PMID- 17460533 TI - Systematic review of the diagnostic accuracy of 201Tl single photon emission computed tomography in the detection of recurrent glioma. AB - OBJECTIVE: To determine the diagnostic accuracy of 201Tl SPECT in the detection of tumour recurrence in patients with previous radiotherapy for supratentorial glioma. METHODS: The databases of PubMed and Embase were searched for relevant studies. Two reviewers independently selected and extracted data on study characteristics, quality and accuracy of studies. Studies were included if they comprised at least six eligible patients who underwent 201Tl SPECT (index test) and in whom (histo)pathological confirmation (reference test) of the suspected brain lesion was obtained. Because of the methodological and statistical heterogeneity of the included studies, a quantitative meta-analysis was not performed. Instead, for every individual study, the sensitivity, specificity and diagnostic odds ratio of 201Tl SPECT was calculated. RESULTS: Eight studies met the inclusion criteria. Only one was considered of high methodological quality. Methodological limitations referred most notably to blinding and patient selection. The diagnostic odds ratio was greater than 1 in all studies included, with a broad range (2-351), and relatively wide 95% confidence intervals. The sensitivity of 201Tl SPECT ranged from 0.43 to 1.00, and the specificity from 0.25 to 1.00. CONCLUSION: 201Tl SPECT seems a valuable method in the detection of tumour recurrence in patients treated with radiotherapy for supratentorial glioma. However, the evidence is not very robust because of the low quality and high heterogeneity of the studies included. Future studies are warranted to further explore the diagnostic potential of 201Tl SPECT, and to determine optimum thresholds for the detection of glioma recurrence. PMID- 17460534 TI - The role of cardiovascular imaging techniques in the assessment of patients with acute chest pain. AB - Chest pain is the most common presenting symptom of coronary artery disease. The assessment and appropriate management of patients with acute chest pain and non diagnostic electrocardiograms (ECGs) remain a continuing clinical problem, with major logistic and financial implications for health-care providers. Cardiovascular imaging is at the forefront of health care, experiencing rapid changes over the recent years, particularly with the use of advanced medical technologies. Imaging techniques like acute rest myocardial perfusion imaging (MPI), echocardiography, electron beam computed tomography (CT), cardiac magnetic resonance imaging (MRI) and multi-detector CT (MDCT) have been used recently in the evaluation and triage of patients with chest pain in addition to the conventional investigations such as ECGs and cardiac biomarkers in the chest pain units. The annual potential cost savings, by incorporating the routine use of acute rest MPI in patients with low-to-moderate risk and non-diagnostic ECGs are substantial. The high negative predictive value of a normal resting MPI in patients with chest pain for myocardial infarction and future cardiac events is well established. Echocardiography is also considered to be useful but the technique is operator dependent and at present there is insufficient data to support its use. Cardiac MRI is expensive and time consuming and there is insufficient diagnostic and prognostic data to make it suitable for chest pain patients at present. There has been increasing interest in MDCT recently, especially with the advent of 64-slice CT but the sensitivity and specificity in chest pain patients are no better than MPI so far. PMID- 17460535 TI - Limitations of the HMPAO SPECT appearances of occipital lobe perfusion in the differential diagnosis of dementia with Lewy bodies. AB - OBJECTIVE: To assess the utility of the appearances of occipital lobe perfusion on HMPAO SPECT in the diagnosis of dementia with Lewy bodies (DLB) using the 123I FP-CIT findings as the diagnostic 'gold standard'. METHODS: Eighty-four consecutive patients underwent both HMPAO SPECT and 123I-FP-CIT as part of their routine investigations for suspected DLB. RESULTS: Thirty-nine of the 84 FP-CIT scans were abnormal indicating a prevalence of 44% of patients with DLB in this series. In those patients classified as DLB, 28% of HMPAO SPECT scans demonstrated occipital hypoperfusion. In those patients with a dementia other than DLB 31% of patients demonstrated occipital hypoperfusion (P=0.8). CONCLUSION: Occipital lobe hypoperfusion as demonstrated by HMPAO SPECT in patients with suspected Lewy body dementia does not appear to be able to either rule in, or rule out, the diagnosis of DLB. PMID- 17460536 TI - Prediction of mortality in patients with coronary artery disease undergoing vasodilator stress testing: a comparison between 99mTc-tetrofosmin and 99mTc sestamibi. AB - PURPOSE: To compare the prognostic ability of the imaging agents 99mTc-sestamibi versus 99mTc-tetrofosmin to predict mortality outcomes in patients with documented coronary artery disease and undergoing vasodilator stress testing. MATERIALS AND METHODS: The study included 2147 consecutive patients who underwent rest and stress single photon emission computed tomographic (SPECT) examination with either 99mTc-sestamibi (n=1128) or 99mTc-tetrofosmin (n=1019). Information relating to all-cause death and cardiovascular death was collected over a 4-year study period. Unadjusted Kaplan-Meier estimates were compared for the two imaging agents. Cox proportional hazard models were examined to determine the incremental contribution of SPECT sum stress score (SSS) and the imaging agent after adjusting for clinical and demographic characteristics. Additionally, the interaction between SSS and agent was examined to determine if the effect of SSS on prognosis was different for the two agents. RESULTS: Vasodilator agents were used for stress testing in all patients who received 99mTc-tetrofosmin and 99mTc sestamibi. Despite differences in patient risk factors Kaplan-Meier estimates were similar for the two groups of patients. Resulting P-values for differences between models for the end points of (1) death from any cause and (2) cardiovascular death showed that SSS combined with clinical index was significantly better than a model that adjusted for only baseline characteristics (P<0.0001 for both endpoints). The addition of imaging agent (99mTc-tetrofosmin or 99mTc-sestamibi) to the model containing both SSS and the clinical characteristics did not show further significant improvement (P=0.62, P=0.96 for death from any cause and cardiovascular death, respectively). CONCLUSION: The type of clinically available 99mTc-labelled myocardial perfusion agents did not affect interpretation of results for prognostic assessment. PMID- 17460537 TI - Potential role of FDG PET in the setting of diabetic neuro-osteoarthropathy: can it differentiate uncomplicated Charcot's neuroarthropathy from osteomyelitis and soft-tissue infection? AB - BACKGROUND: This paper is based on the results from an ongoing prospective trial designed to investigate the usefulness of FDG PET in the complicated diabetic foot. AIM: To investigate the potential utility of FDG PET imaging in the setting of acute neuropathic osteoarthropathy (Charcot's foot). PATIENTS AND METHODS: A total of 63 patients, in four groups, were evaluated. The groups were: (A) 17 patients with a clinical diagnosis of Charcot's neuroarthropathy (11 men, six women; mean age: 59.4+/-8.6 years); (B) 21 patients with uncomplicated diabetic foot (16 men, five women; mean age: 63+/-10 years); (C) 20 non-diabetic patients with normal lower extremities (12 men, eight women; mean age 54+/-19 years); and (D) five patients with proven osteomyelitis secondary to complicated diabetic foot (three men, two women; mean age: 61.2+/-13.9 years). Five patients in group A had foot ulcer and intermediate to high degree of suspicion for superimposed osteomyelitis. Each subject underwent FDG PET imaging of the lower extremities in addition to MRI and the findings were compared with the final diagnostic outcome based on histopathology and clinical follow-up. The images were examined visually for focal abnormalities. Regions of interest were assigned to the sites of abnormal FDG uptake for calculating maximum standardized uptake value (SUVmax). Two important clinical decision-making issues were explored: (1) whether FDG PET shows a definitive uptake pattern in Charcot's neuroarthropathy and if so whether that could be utilized to differentiate it from other complicated forms of diabetic foot like osteomyelitis and cellulitis, which is frequently a diagnostic challenge in this clinical setting; and (2) how accurate FDG PET is in detection soft tissue infection in patients with Charcot's foot. These issues were examined by utilizing FDG PET findings along with MRI results in the same patient. RESULTS: We observed a low degree of diffuse FDG uptake in the Charcot's joints. This was clearly distinguishable from the normal joints. The SUVmax in the Charcot's lesions varied from 0.7 to 2.4 (mean, 1.3+/-0.4) while those of midfoot of the normal control subjects and the uncomplicated diabetic foot ranged from 0.2 to 0.7 (mean 0.42+/-0.12) and from 0.2 to 0.8 (mean 0.5+/-0.16), respectively. The only patient with Charcot's foot with superimposed osteomyelitis had an SUVmax of 6.5. The SUVmax of the sites of osteomyelitis as a complication of diabetic foot was 2.9-6.2 (mean: 4.38+/-1.39). Unifactorial analysis of variance test yielded a statistical significance in the SUVmax between the four groups (P<0.01). The SUVmax between the normal control groups and the uncomplicated diabetic foot was not statistically significant by the Student's t-test (P>0.05). In the setting of concomitant foot ulcer FDG PET accurately ruled out osteomyelitis. Overall sensitivity and accuracy of FDG PET in the diagnosis of Charcot's foot was 100 and 93.8%, respectively; and for MRI were 76.9 and 75%, respectively. FDG PET showed foci of abnormally enhanced uptake in the soft tissue which was suggestive of inflammation in seven cases (43.75%) which were proven pathologically to be secondary to infection. In only two of these cases the features of soft tissue infection were noted on the magnetic resonance images. CONCLUSION: The results support a valuable role of FDG PET in the setting of Charcot's neuroarthropathy by reliably differentiating it from osteomyelitis both in general and when foot ulcer is present. PMID- 17460538 TI - Role of 18F-dopa PET/CT imaging in the management of patients with 111In pentetreotide negative GEP tumours. AB - PURPOSE: To assess whether 18F-dopa PET/CT is able to provide information relevant in changing the clinical management of patients with gastro enteropancreatic (GEP) tumours where there is negative or inconclusive conventional radiological imaging (ultrasound (US), computed tomography (CT) and magnetic resonance imaging (MRI)) and 111In-pentetreotide scintigraphy. MATERIALS AND METHODS: From January 2005 to October 2006, 84 patients with clinical and biochemical suspicion of GEP tumours were investigated by US and CT scans, MRI and 111In-pentetreotide scintigraphy. In 13/84 (15.4%) both conventional radiological imaging and 111In-pentetreotide scintigraphy provided negative or inconclusive findings, and patients were referred for 18F-dopa PET/CT imaging. Each patient received 5.3 MBq x kg(-1) 18F-dopa intravenously, and imaged 60 min later using a hybrid PET/CT scanner. RESULTS: 18F-dopa PET/CT detected the primary tumour in all 13 patients (size range, 7-26 mm, mean, 18 mm; SUVmax range, 2.3-16.3, mean, 5.7) and further 12 unsuspected lesions (size range, 12-23 mm, mean 17; SUVmax range 2.8-12.7, mean 4.6). Confirmation of the PET/CT findings was obtained in all patients from histopathological analysis of tissue obtained after surgery and/or biopsy. All the 18F-dopa-positive primary lesions were confirmed as being the primary tumour at histology, whereas of the other 12 unsuspected 18F-dopa-positive lesions, 11 were found to be metastatic deposits and one due to unspecific inflammation (one false positive result). Notably, the results of 18F-dopa PET/CT imaging changed the clinical management in 11/13 patients (84%). CONCLUSIONS: Our preliminary results suggest that 18F-dopa PET/CT has a promising role in GEP patients with negative or inconclusive findings at conventional radiological imaging and 111In-pentetreotide scintigraphy. The findings were helpful in biopsy guidance and played a major role in changing the management of those patients. PMID- 17460539 TI - Dynamic antral scintigraphy following solid and liquid meals in healthy human subjects. AB - AIMS: Dynamic antral scintigraphy (DAS), a non-invasive technique for the assessment of post-prandial gastric contractions, has been used to demonstrate abnormal contractility in several clinical conditions. The objective of the present study was to assess differences between solid and liquid meals regarding gastric contractions in healthy volunteers using DAS. METHODS: Ten healthy male volunteers were studied after ingesting solid or liquid meals [approximately 1670 kJ (approximately 400 kcal)] labelled with 99mTc phytate and administered in a random order. Gastric images were acquired for 120 min for gastric emptying half time (T1/2) measurement. Dynamic (1 frame x s(-1)) images of the gastric antrum were acquired at 30 min intervals for 4 min for the assessment of antral contractility. RESULTS: Gastric emptying T1/2 values for solid and liquid meals were similar (58.1+/-19.06 min vs. 69.4+/-6.76 min; P=0.13). For the solid meal, average values for both frequency (3.08+/-0.15 cycles x min(-1) vs. 2.78+/-0.18 cycles x min(-1); P=0.003) and amplitude (33.94+/-5.2% variation vs. 24.09+/ 7.37% variation; P=0.002) of antral contractions were significantly higher than those obtained with the liquid meal. For either of the test meals, none of the antral contractility variables correlated with gastric emptying T1/2. CONCLUSIONS: Dynamic antral scintigraphy is capable of detecting differences between solid and liquid meals concerning post-prandial gastric antral contractions under physiological conditions. The frequency and amplitude of gastric antral contractions after a solid meal are greater than after an exclusively liquid meal of similar calorie content, in spite of lack of a difference regarding gastric emptying. PMID- 17460540 TI - A novel iterative method for lesion delineation and volumetric quantification with FDG PET. AB - OBJECTIVES: The determination of lesion boundaries on FDG PET is difficult due to the point-spread blurring and unknown uptake of activity within a lesion. Standard threshold-based methods for volumetric quantification on PET usually neglect any size dependence and are biased by dependence on the signal-to background ratio (SBR). A novel, model-based method is hypothesized to provide threshold levels independent f the SBR and to allow accurate measurement of volumes down to the resolution of the PET scanner. METHODS: A background subtracted relative-threshold level (RTL) method was derived, based on a convolution of the point-spread function and a sphere with diameter D. Validation of the RTL method was performed using PET imaging of a Jaszczak phantom with seven hollow spheres (D=10-60 mm). Activity concentrations for the background and spheres (signal) were varied to obtain SBRs of 1.5-10. An iterative procedure was introduced for volumetric quantification, as the optimal RTL depends on a priori knowledge of the volume. The feasibility of the RTL method was tested in two patients with liver metastases and compared to a standard method using a fixed percentage of the signal. RESULTS: Phantom data validated that the theoretically optimal RTL depends on the sphere size, but not on the SBR. Typically, RTL=40% (D=15-60 mm), and RTL>50% for small spheres (D<12 mm). The RTL method is better applicable to patient data than the standard method. CONCLUSIONS: Based on an iterative procedure, the RTL method has been shown to provide optimal threshold levels independent of the SBR and to be applicable in phantom and in patient studies. It is a promising tool for lesion delineation and volumetric quantification of PET lesions. PMID- 17460541 TI - High-density materials do not always induce artifacts on PET/CT: what is responsible for the difference? AB - OBJECTIVE: PET/CT often show increased uptake at sites of high-density materials. However, some materials seldom demonstrate increased uptake on PET/CT, such as the materials used in hip prostheses. We hypothesized that the motion of materials may be crucial for such artifacts. Here, we present representative cases, and validate our hypothesis based on the results of phantom studies. METHODS: A standard cylinder, 20 cm in diameter, was filled with approximately 37 MBq of 18F-based activity, and a pacemaker was attached to the side of the cylinder. This phantom was placed on the bed with the pacemaker side facing the scanner. PET scans were performed using a Biograph LSO DUO. CT scans were performed first for transmission scans, followed by acquisition of emission scans. The phantom was first scanned (protocol 1). The phantom was then moved about 2 cm closer to the distal edge of the bed just after transmission CT scan, and the emission scan was performed (protocol 2). RESULTS: Homogenous uptake was seen in the cylinder in protocol 1, and there was no visible uptake at the site of the pacemaker. In contrast, a clear hotspot was seen at the site of the pacemaker in protocol 2. The uptake in the cylinder was inhomogeneous; that on the pacemaker side of the cylinder was low, while that on the opposite side was high. CONCLUSIONS: High-density materials do not show false increased uptake without motion on PET/CT. Motion of these materials surrounded by radioactive organs may play an important role in inducing false increased uptake on PET/CT. PMID- 17460542 TI - Radiosynthesis of 18F-(R8,15,21, L17)-vasoactive intestinal peptide and preliminary evaluation in mice bearing C26 colorectal tumours. AB - BACKGROUND: Radiolabelled vasoactive intestinal peptide (VIP) and its analogues have shown their potential as imaging agents for diagnosing tumours expressing VIP receptor. However, the fast proteolytic degradation in vivo has limited their clinical use. AIM: To prepare the 18F-labelled (R8,15,21, L17)-VIP analogue in a convenient way and to evaluate its potential as an imaging agent for VIP receptor positive tumours. METHODS: Radiolabelled (R8,15,21, L17)-VIP was obtained by conjugation with N-succinimidyl 4-([18F]fluoromethyl) benzoate and purified by HPLC. Radiochemical purity and specific radioactivity were measured by analytical HPLC. In-vitro stability of the product was carried out in HSA solution and analysed by HPLC. Biodistribution study was carried out in mice bearing C26 colorectal tumours. RESULTS: 18F-(R8,15,21, L17)-VIP was obtained in greater than 99% radiochemical purity within 60 min in decay-for-corrected radiochemical yields of 21.8+/-4.7% (n=5) and a specific activity of 17.76 GBq x mumol(-1) at the end of synthesis (EOS). Results of in-vitro studies demonstrated a high stability in human serum albumin (HSA) solution. Biodistribution data showed a rapid blood clearance and specific binding towards receptor-positive tumours. CONCLUSION: 18F-(R8,15,21, L17)-VIP was prepared by a convenient method. Preliminary biodistribution results showed its potential for imaging tumours over expressing VIP receptors and encouraged further investigation. PMID- 17460543 TI - Long-term recurrence of thyroid cancer after thyroid remnant ablation with 1.1 and 3.7 GBq radioiodine. PMID- 17460545 TI - The discovery of new coding alleles of human CYP26A1 that are potentially defective in the metabolism of all-trans retinoic acid and their assessment in a recombinant cDNA expression system. AB - OBJECTIVES: Retinoic acid (RA) is a critical regulator of gene expression during embryonic development and in the maintenance of adult epithelial tissues. This study was undertaken to identify genetic polymorphisms of CYP26A1 which might affect these processes. We sequenced CYP26A1 in racially diverse individuals and assessed the metabolism of retinoic acid by newly identified coding alleles of CYP26A1 in a recombinant system. METHODS: CYP26A1 was sequenced in 24 Caucasians, 24 African-Americans, 24 Asians, and 20 individuals of unknown racial origin. cDNA constructs for wild-type and coding alleles of CYP26A1 were constructed in a pcDNA3.1 expression vector and expressed in Cos-1 cells. A FLAG tag at the C terminal end of the cDNA was used to quantitate the recombinant CYP26A1 proteins. RESULTS: A total of 13 single nucleotide polymorphisms (SNPs) were identified in CYP26A1. Three SNPs produced coding changes: R173S, F186L, and C358R. These alleles were termed as CYP26A1*2, CYP26A1*3, and CYP26A1*4, respectively, by the Human Cytochrome P450 (CYP) Allele Nomenclature Committee at http://www.cypalleles.ki.se/. Wild type CYP26A1 protein metabolized all-trans retinoic acid (at-RA) to 4-oxo-RA, 4-OH-RA as well as water-soluble metabolites. CYP26A1.3 (F186L) and CYP26A1.4 (C358R) allelic proteins exhibited significantly lower metabolism (40-80%) of at-RA than wild-type CYP26A1.1 protein. CONCLUSION: This is the first study to identify coding alleles of CYP26A1. Two coding alleles, CYP26A1*3 and CYP26A1*4, are predicted to be defective based on the metabolism of at-RA by the recombinant proteins. These studies suggest the need for future clinical studies of polymorphisms of CYP26A1 in embryonic development. PMID- 17460546 TI - Description of a novel polymorphic gene encoding for arylamine N acetyltransferase in the rhesus macaque (Macaca mulatta), a model animal for endometriosis. AB - OBJECTIVES: Case-control studies have previously associated polymorphisms in the gene encoding the xenobiotic metabolizing enzyme arylamine N-acetyltransferase 2 (NAT2) with endometriosis, a common multifactorial disease in women. These studies, however, have been problematic on methodological grounds and their results are inconclusive. To better understand the possible relationship between the NAT2 gene and endometriosis, we characterized its homologue in the rhesus macaque, an animal model for the disease. METHODS: Human NAT2-specific primers were used to isolate orthologous gene sequences from four unrelated rhesus macaques of the same colony. Recombinant proteins were expressed in mammalian cells and analysed for their ability to acetylate NAT substrates and bind anti NAT antibodies. RESULTS: A polymorphic gene, showing 94% identity to human NAT2, was identified in the rhesus macaque. Its two characterized alleles, designated (MACMU)NAT2*1 and (MACMU)NAT2*2, were differentiated by one synonymous (C(624)T) and one nonsynonymous (G(691)A) polymorphism, the latter causing a Val(231)Ile substitution. The recombinant (MACMU)NAT2 protein was not recognized by anti (HUMAN)NAT1 antibody, but reacted with antibodies against (HUMAN)NAT2 or the active site of NAT. Rhesus NAT2 provided relatively high acetylation activity with p-anisidine, lower activity with procainamide, sulphamethazine or 5 aminosalicylate and poor activity with p-aminobenzoic acid. Differences in the activities of the two allozymes were evident with most substrates. CONCLUSIONS: A polymorphic homologue of human NAT2 was characterized in the rhesus macaque, to facilitate investigations of the postulated involvement of this isoenzyme in the toxicogenetics of endometriosis. PMID- 17460547 TI - Pharmacogenetics of the 5-lipoxygenase biosynthetic pathway and variable clinical response to montelukast. AB - OBJECTIVE: Interindividual clinical response to leukotriene modifiers is highly variable, and less efficacious than inhaled corticosteroids in treating asthma. Genetic variability in 5-lipoxygenase biosynthetic and receptor pathway gene loci may influence cysteinyl-leukotriene production and subsequent response to leukotriene modifiers. METHODS: Using data from two clinical trials of 12-week duration, post-hoc analyses were performed in 174 patients randomized to montelukast. Associations between polymorphisms in 10 candidate genes (ALOX5, ALOX5AP, LTC4S, CYSLTR1, CYSLTR2, PLA2G4A, CYP2C9, CYP3A4, ADRB2, and NR3C1) and response to montelukast were modeled using change in morning peak expiratory flow and forced expiratory volume in 1 s (FEV1) to define the response phenotype. RESULTS: In our sample, eight out of 25 markers in 10 candidate genes were statistically associated with response to montelukast, with an estimated proportion of false discoveries of 16%. The strongest statistical evidence of clinically relevant pharmacogenetic effects peak expiratory flow were identified in CYSLTR2 (rs91227 and rs912278; P=0.02 and P=0.02, respectively) and ALOX5 (rs4987105 and rs4986832; P=0.01 and P=0.01, respectively). Patients with these variant genotypes, found in roughly 10-13% of patients, had an 18-25% improvement in peak expiratory flow. In contrast, the majority of patients with the wild-type alleles had only a marginal (8-10%) improvement. CONCLUSIONS: The overall mean response to montelukast may be skewed towards a response phenotype by a small subset (<15%) of asthma patients. CYSLTR2 and ALOX5 polymorphisms may predispose a minority of individuals to excessive cysteinyl-leukotriene concentrations, yielding a distinct asthma phenotype most likely to respond to leukotriene modifier pharmacotherapy. These findings require replication to establish validity and clinical utility. PMID- 17460548 TI - Platelet 12-lipoxygenase Arg261Gln polymorphism: functional characterization and association with risk of esophageal squamous cell carcinoma in combination with COX-2 polymorphisms. AB - BACKGROUND: Aberrant arachidonic acid metabolism by 12-lipoxygenase (12-LOX) and cyclooxygenase-2 (COX-2) has been implicated in human carcinogenesis. Inherited polymorphisms in 12-LOX and COX-2 contributed to differential expression or activity of these enzymes might confer interindividual susceptibility to cancer. OBJECTIVE: To examine the functional significance of 12-LOX 261 Arg> Gln polymorphism and its association, alone and in combination with COX-2 -1195G > A and -765G > C polymorphisms, with risk of developing esophageal squamous cell carcinoma (ESCC). METHODS: The platelet 12-LOX activity was measured by quantifying 12-HETE in the lipoxygenation reaction. Genotypes of 12-LOX261Arg>Gln and COX-2 -1195G>A and -765G>C polymorphisms were determined in a case-control study consisting of 1026 patients and 1270 controls. Associations with the risk of ESCC were estimated by logistic regression. RESULTS: Subjects with the 12-LOX Gln/Gln genotype had higher platelet 12-LOX activity (mean+/-SEM nmol/mg/min) than those with the Arg/Arg genotype (0.405+/-0.047 [n=10] versus 0.136+/-0.022 [n=6]; P=0.001). Genotyping data showed that the 12-LOX Gln/Gln genotype was associated with increased risk of developing ESCC (odds ratio [OR]=1.42, 95% confidence interval [CI]=1.12-1.81), compared with the Arg/Arg genotype adjusted for sex, age, and smoking. An increased risk of ESCC was also associated with the COX-2 -1195GA (OR=1.34, 95% CI=1.08-1.68; P=0.008), -1195AA (OR=1.72, 95% CI=1.35 2.20; P=<0.001), and -765GC (OR=2.24, 95% CI=1.59-3.16; P<0.001) genotypes. Furthermore, a multiplicative interaction between the 12-LOX Gln/Gln and COX-2 1195AA or -765GC genotype in intensifying risk of ESCC was observed, with the ORs for the presence of both 12-LOX Gln/Gln and COX-2 -1195AA or -765GC genotypes being 3.21 (95% CI=1.93-5.34) and 3.33 (95% CI=1.59-6.98). A multiplicative interaction between the -765GC genotype and smoking was also evident (OR=4.45, 95% CI=2.71-7.29). CONCLUSION: These observations suggest that inherited polymorphisms in arachidonic acid-metabolizing enzymes, which result in heightened gene expression or enzymatic activity, may confer host susceptibility to ESCC. PMID- 17460549 TI - A splice site polymorphism in the G-protein beta subunit influences antidepressant efficacy in depression. AB - OBJECTIVE: Recent evidence suggests that signalling cascades located downstream of monoamine receptors are altered following antidepressant treatment. Our objective was to investigate whether genetic polymorphisms in genes involved in these signalling cascades influenced antidepressant efficacy. METHODS: Polymorphisms in the G-protein beta subunit GNB3, the cAMP response element binding protein 1 gene (CREB1), the brain derived neurotrophic factor (BDNF) and CREB binding protein (CREBBP) were studied in well characterised unipolar (n=166) and early onset (n=102) depressive populations and correlated with treatment response. RESULTS: The GNB3 C825T polymorphism, which results in a 41 amino acid deletion, was significantly associated with lack of remission (OR=0.18, P=0.02) and lack of response (OR=0.26, P=0.03) following 2nd switch treatment. A cytosine deletion 16 base pairs from the start of exon 8 in CREB1 was found more frequently in remitters and responders to 2nd switch antidepressant drug therapy, although these differences failed to reach significance. Polymorphisms detected BDNF (G196A) and CREBBP (T651 C) did not appear to influence antidepressant response. CONCLUSIONS: These results suggest that inheritance of the GNB3C825T allele may significantly influence antidepressant response and emphasises the potential importance of polymorphisms in genes in signalling cascades activated by commonly prescribed antidepressants. PMID- 17460550 TI - Common ABCB1 polymorphisms are not associated with multidrug resistance in epilepsy using a gene-wide tagging approach. AB - P-glycoprotein, the product of the ABCB1 gene, is a proposed mechanism of pharmacoresistance in epilepsy. Previous attempts to correlate the ABCB1 C3435T SNP, or a three-SNP haplotype containing C3435T with epilepsy pharmacoresistance have produced discordant findings. We analysed these single nucleotide polymorphisms (SNPs), plus a more comprehensive set of tagging SNPs describing common variation in ABCB1 in a case-control study. No significant association of C3435T (P=0.55), the three-SNP haplotype (lowest P=0.14) or any gene-wide tagging SNP (lowest P=0.17) with multidrug resistance in epilepsy was identified. Meta analysis of studies using the same definition of multidrug resistance (n=1064) also demonstrated no significant association of C3435T with multidrug resistance (P=0.31). These findings suggest that C3435T is unlikely to be a marker for epilepsy multidrug resistance. In addition, no evidence for a role of other common ABCB1 polymorphisms was found using a potentially more powerful gene-wide tagging approach. PMID- 17460551 TI - Polymorphisms predicted to alter function in prostaglandin E2 synthase and prostaglandin E2 receptors. AB - BACKGROUND AND OBJECTIVE: Prostaglandin synthesis is the primary target of aspirin and other nonsteroidal antiinflammatory drugs, and thus is a pathway of major interest to pharmacology, pharmacogenetics, and epidemiology. Several lines of evidence implicate prostaglandin E2 in carcinogenesis; this study aimed to identify genetic variants in genes related to prostaglandin E2 synthesis and signaling. METHODS: We resequenced the coding regions of human prostaglandin E2 synthase (PGES), and prostaglandin E2 receptors EP1, EP2, and EP4 in 48 African Americans and 47 Caucasians. RESULTS AND CONCLUSIONS: We identified 23 variants, 6 of which cause amino acid changes. The non-synonymous polymorphisms in PGES, EP1, and EP2 were present only among African-Americans; both populations carried non-synonymous polymorphisms in EP4. We used two sequence homology-based programs, SIFT and PolyPhen, to predict the impact of these polymorphisms. These programs predicted that the amino-acid changes p.Phe119Val in EP1, p.Ala44Glu in EP2, and possibly p.Val7Glu in PGES, p.Thr176Ile in EP4 and p.Gly420Asp in EP4 are likely to affect protein function. Thus, these variants may be relevant for inflammatory conditions, carcinogenesis, and pharmacogenetics. PMID- 17460552 TI - Fish oil treatment for kidney transplant recipients: a meta-analysis of randomized controlled trials. AB - BACKGROUND: Calcineurin inhibitors have adverse effects that contribute to nephrotoxicity and cardiovascular risk profile, and these may be reduced by administration of fish oil. The aim of this review was to assess the benefits and harms of fish oil supplementation in kidney transplant recipients on a calcineurin inhibitor-based immunosuppressive regimen. METHODS: The Cochrane Controlled Trials Registry, MEDLINE, and EMBASE were searched for randomized controlled trials of fish oil treatment in kidney transplant recipients on a calcineurin inhibitor-based immunosuppressive regimen. Trials comparing fish oil to both placebo and statins were included. Data were extracted for patient and graft survival, acute rejection, calcineurin inhibitor toxicity, cardiovascular events, adverse effects, compliance, renal function, blood pressure, and lipid profile. Dichotomous outcomes were reported as relative risk and continuous outcome measures as weighted mean differences (WMD), with 95% confidence intervals. RESULTS: Sixteen suitable trials were analyzed. Fish oil treatment was associated with a lower diastolic blood pressure (WMD 4.5 mmHg, P=0.004) and higher high-density lipoprotein (HDL) cholesterol (WMD 0.12 mmol/L, P=0.01) but did not affect the other outcomes. Fishy aftertaste and gastrointestinal upset were common but did not result in significant dropout. Fish oil effects on lipids were not significantly different than low-dose statins. CONCLUSION: There is insufficient evidence from currently available randomized controlled trials to recommend fish oil therapy to improve renal function, rejection rates, and patient or graft survival. Improvements in HDL cholesterol and diastolic blood pressure were too modest to recommend routine use. PMID- 17460553 TI - Activated circulating dendritic cells after hematopoietic stem cell transplantation predict acute graft-versus-host disease. AB - BACKGROUND: Dendritic cells (DC) are central to the development of acute graft versus-host disease (GVHD) following allogeneic hematopoietic stem cell transplantation (alloHSCT). We hypothesized that DC activation status determines the severity of GVHD and that activated DC may be detected in the circulation prior to clinical presentation of GVHD. METHODS: Following transplant, blood samples were obtained twice weekly from alloHSCT patients. Myeloid (CD11c+) and plasmacytoid (CD123hi) DC were enumerated by flow cytometry, and activated myeloid DC were identified using the CMRF-44 monoclonal antibody. RESULTS: Of 40 alloHSCT patients, 26 developed acute GVHD. Severity of GVHD was associated with low total blood DC counts (P=0.007) and with low myeloid and plasmacytoid DC numbers (P=0.015 and 0.003). The CMRF-44 antigen was expressed on blood CD11c+ DC in all cases prior to GVHD onset, whereas of the 14 patients without GVHD, seven had no CMRF-44+ CD11c DC. Patients with CMRF-44+ CD11c+ DC in more than 20% of samples were more likely to subsequently develop acute GVHD (P=0.001, odds ratio=37.1), while patients who developed grade 2-4 GVHD had prior higher percentages of CMRF-44+ CD11c+ DC compared to grade 0-1 GVHD patients (P=0.001). CMRF-44 expression on >7.9% CD11c+ DC predicted for subsequent development of GVHD with a sensitivity of 87.5% and specificity of 79.2%. CONCLUSIONS: Activation status, as assessed by CMRF-44 antigen expression, of blood CD11c+ DC is highly associated with acute GVHD and these cells may be targets for therapeutic intervention. PMID- 17460554 TI - Pretransplant cellular alloimmunity as assessed by a panel of reactive T cells assay correlates with acute renal graft rejection. AB - BACKGROUND: The panel reactive antibody test (PRA) is an established method for assessing posttransplant risk of immune-mediated graft injury. The panel of reactive T cell assay (PRT) in which transplant candidates' peripheral blood mononuclear cells are tested for reactivity to a panel of allogenic stimulator cells by the IFN-gamma enzyme-linked immunosorbent spot assay analogously assesses the strength of the pretransplant effector-memory alloreactive T cell repertoire. METHODS: PRT assays were performed in 30 kidney transplant candidates and results were correlated with acute rejection (AR). A positive PRT assay was defined as a response to at least 75% of the stimulators tested. RESULTS: A positive pretransplant PRT test was observed in 11 of 30 (37%) patients, and AR within 1 year posttransplantation was seen in 7 of 30 (23%) subjects. Six of the seven (86%) patients with AR were PRT-positive (P=0.01) whereas only one of seven (14%) patients with a PRA greater than 15% had AR. The mean pretransplant PRT percentage was 40% for patients with no AR versus 81% for patients with AR (P=0.01). Estimated glomerular filtration rate (mL/min/1.73 m2) showed a trend towards a lower value in PRT-positive (48+/-15) versus PRT-negative (55+/-13) individuals. CONCLUSIONS: The data suggest that pretransplant PRT screening can identify patients at risk for posttransplant cellular immune mediated graft injury despite the absence of humoral allosensitization. Once confirmed by larger prospective trials, PRT screening could be used to guide clinical decision-making with regard to choosing donor organs and individualizing immunosuppression regimens. PMID- 17460555 TI - Hepatitis C virus infection and kidney transplantation: predictors of patient and graft survival. AB - BACKGROUND: The effect of hepatitis C virus (HCV) infection on patients undergoing kidney transplantation (KTx) is uncertain. This study aimed to evaluate the outcomes of our HCV+/end-stage renal disease (ESRD) patient population based on the therapeutic option including KTx or continuation in dialysis. METHODS: KTx performed at Virginia Commonwealth University Hospital between January 2000 and December 2004 were tracked prospectively. Forty-three out of a total of 394 KTx patients included in the analysis were HCV+. A group of 52 contemporaneous HCV+/ESRD patients listed, but never transplanted, was also analyzed. HCV-negative transplanted patients were used as the control group. RESULTS: Patient survival posttransplantation was 81.4% and 68.5% at 1 and 3 years in the HCV+ group, and 97.1% and 92.9% at 1 and 3 years in the HCV- group, respectively (P=0.001). Graft survival was 81.2% and 64.1% at 1 and 3 years in the HCV+ group, and 93.2% and 84.1% at 1 and 3 years posttransplantation in the HCV- group (P=0.01). Univariate analysis identified Knodell score as a predictor of mortality in HCV+ patients (P=0.04). Cox proportional hazards multivariate analysis identified deceased donor (P=0.02), previous kidney transplant (P=0.007), pretransplant diabetes (P=0.05), and Knodell Score (P=0.012) as predictors of patient mortality. Patient survival was superior in HCV+ patients undergoing KTx versus remaining on dialysis. CONCLUSIONS: Patients with ESRD/HCV+ benefit from KTx without achieving the excellent survival of HCV-/ESRD patients. Liver biopsy is a useful tool to identify advanced liver disease at pretransplantation time. PMID- 17460556 TI - Rates and risk factors for nonadherence to the medical regimen after adult solid organ transplantation. AB - BACKGROUND: Despite the impact of medical regimen nonadherence on health outcomes after organ transplantation, there is mixed and conflicting evidence regarding the prevalence and predictors of posttransplant nonadherence. Clinicians require precise information on nonadherence rates in order to evaluate patients' risks for this problem. METHODS: A total of 147 studies of kidney, heart, liver, pancreas/kidney-pancreas, or lung/heart-lung recipients published between 1981 and 2005 were included in a meta-analysis. Average nonadherence rates were calculated for 10 areas of the medical regimen. Correlations between nonadherence and patient psychosocial risk factors were examined. RESULTS: Across all types of transplantation, average nonadherence rates ranged from 1 to 4 cases per 100 patients per year (PPY) for substance use (tobacco, alcohol, illicit drugs), to 19 to 25 cases per 100 PPY for nonadherence to immunosuppressants, diet, exercise, and other healthcare requirements. Rates varied significantly by transplant type in two areas: immunosuppressant nonadherence was highest in kidney recipients (36 cases per 100 PPY vs. 7 to 15 cases in other recipients). Failure to exercise was highest in heart recipients (34 cases per 100 PPY vs. 9 to 22 cases in other recipients). Demographics, social support, and perceived health showed little correlation with nonadherence. Pretransplant substance use predicted posttransplant use. CONCLUSIONS: The estimated nonadherence rates, overall and by transplant type, allow clinicians to gauge patient risk and target resources accordingly. Nonadherence rates in some areas--including immunosuppressant use--appear unacceptably high. Weak correlations of most patient psychosocial factors with nonadherence suggest that attention should focus on other classes of variables (e.g., provider-related and systems-level factors), which may be more influential. PMID- 17460557 TI - Leukopenia complicates cytomegalovirus prevention after renal transplantation with alemtuzumab induction. AB - BACKGROUND: Cytomegalovirus (CMV) is a major cause of morbidity after transplantation. Valganciclovir (VGCV) is commonly utilized for CMV prophylaxis but can cause leukopenia, with risk compounded by the use of myelosuppressive immunosuppression. By utilizing a preemptive therapeutic strategy with VGCV targeted only toward patients at risk for developing CMV disease, the rate and extent of leukopenia may be reduced. METHODS: VGCV prophylactic and preemptive strategies were compared in renal transplant recipients receiving alemtuzumab induction and prednisone-free maintenance with tacrolimus and mycophenolate mofetil (MMF). Patients were risk-stratified by CMV serologic status. All donor seropositive/recipient seronegative (D+/R-) patients, February 2002-January 2004 (n=32), received prophylaxis with VGCV 450 mg daily for 3 months. Outcomes of D+/ /R+ patients were compared. Patients in the first cohort, February 2002-October 2002 (n=61), received prophylaxis as described. In the second cohort, October 2002-January 2004 (n=110), patients were monitored by quantitative CMV-polymerase chain reaction (PCR) every 2 weeks for 3 months. If the CMV load was above laboratory threshold, VGCV 450 mg daily was initiated for 1 month or until viremia cleared. RESULTS: Comparing preemptive therapy versus prophylaxis in D+/ /R+ patients, there was a lower incidence of leukopenia (67% vs. 82%, P=0.039) and trend toward less granulocyte-colony stimulating factor (G-CSF) use (24% vs. 33%, P=0.196), but higher CMV disease rate (15% vs. 3%, P=0.02). One limitation was strategy compliance: 41% (7 of 17) of preemptive patients who developed CMV missed at least 1 CMV-PCR before diagnosis. One-year patient (98.2% vs. 98.4%) and death-censored graft (100% vs. 98.4%) survival was similar. CONCLUSIONS: Antiviral toxicity may be decreased with preemptive therapy, but effectiveness for CMV prevention seems dependent upon monitoring compliance. PMID- 17460558 TI - Kidney transplantation with sirolimus and mycophenolate mofetil-based immunosuppression: 5-year results of a randomized prospective trial compared to calcineurin inhibitor drugs. AB - BACKGROUND: We report the 5-year outcomes from a randomized prospective trial in primary adult renal allograft recipients, designed to evaluate calcineurin inhibitor (CNI)-free immunosuppression on kidney transplant function. METHODS: Sixty-one patients were randomized to either sirolimus (n=31) or cyclosporine (n=30) after basiliximab induction and mycophenolate mofetil (MMF) with steroids. Sirolimus was concentration controlled at 10-12 ng/mL for at least 6 months. RESULTS: After 5 years, sirolimus-MMF-steroids compared to cyclosporine-MMF steroids provides similar patient survival (87.1 vs. 90%, P=0.681), acute rejection rates (12.9 vs. 23.3%, P=0.22), total cholesterol (209.1 vs. 204.3 mg/dL, P=0.973), urine protein/creatinine ratios (0.398 vs. 0.478 mg/dL, P=0.72), and overall medical and surgical morbidity (P=NS). Although unadjusted patient survival was similar, sirolimus based CNI-free patients had longer death censored graft survival (96.4 vs. 76.7%, P=0.0265), higher glomerular filtration rate (GFR) by the abbreviated Modified Diet in Renal Disease (66.7 vs. 50.7 cc/min, P=0.0075), and fewer graft losses from chronic allograft nephropathy. The Banff chronic scores at two years were strong predictors of 5-year GFR. At 5 years, there were six de novo (three solid organ, three skin) cancers in the CNI group and only two de novo (one skin, one leukemia, no solid organ) cancers in the sirolimus group (P=NS). CONCLUSIONS: This study of low to moderate risk patients demonstrates that excellent 5-year kidney transplant outcomes can be achieved without CNI drugs, when therapeutic drug monitoring of sirolimus is employed. The application of CNI drug avoidance protocols to high-risk recipients (retransplants, highly sensitized, etc.), extrarenal allograft recipients, or alternative drug regimens such as steroid or MMF elimination should be subjected to controlled trials. PMID- 17460559 TI - Extended indication for living donor liver transplantation in patients with hepatocellular carcinoma. AB - BACKGROUND: Liver transplantation is an accepted treatment option for patients with otherwise untreatable hepatocellular carcinoma (HCC). The present study assessed the outcome of living donor liver transplantation (LDLT) under extended selection criteria based on a single-center experience. METHODS: A total of 60 patients who underwent LDLT for HCC were included. Our indication for LDLT included HCC without extrahepatic spread or macroscopic vascular invasion. The size and number of HCC nodules were not limited. Recurrence-free survival rates according to various factors were compared to identify risk factors for recurrence. RESULTS: Forty patients (67%) preoperatively exceeded the Milan criteria. The median follow-up was 437 days (range: 23-1,385 days). The overall 1 and 3-year actuarial survival rates were 88.4 and 68.6%, respectively. HCC recurred in eight patients (14.3%) within a mean follow-up of 288 days; all were patients who exceeded the Milan criteria. The 1-, 2- and 3-year recurrence-free survival rates of patients who fulfilled the Milan criteria were 100%, 100%, and 100%, respectively, whereas those of patients who exceeded the criteria were 83.0%, 74.0%, and 74.0%, respectively. Tumor diameter >5 cm was significantly associated with worse prognosis, but the number of tumors was not. A preoperative des-gamma-carboxy prothrombin value >300 mAU/ml was strongly associated with the high recurrence rate. These two variables were significant in multivariate analysis. CONCLUSIONS: LDLT was shown to offer acceptable results in patients who exceeded the Milan criteria. The indication for LDLT can therefore be expanded beyond the Milan criteria, especially for patients with small multiple tumors <5 cm. PMID- 17460560 TI - Five-year outcome in pediatric patients with mycophenolate mofetil-based renal transplantation. AB - BACKGROUND: Mycophenolate mofetil (MMF) based immunosuppression after renal transplantation has proven to be safe and beneficial for children and adolescents. However, long-term analysis, in particular of pediatric patients, is scarce. PATIENTS: Data of 140 patients receiving MMF versus azathioprine (AZA) in combination with cyclosporine A (CsA) and prednisone without induction were analyzed with a main focus on survival and renal function in long-term follow-up. RESULTS: After 5 years of follow-up, 44 MMF and 20 AZA patients were still on study. Graft survival of intent to treat (ITT) groups was 90.7% for MMF and 68.5% for AZA patients (P<0.001). Cumulative rejection free survival was 51.2% in MMF versus 37.0% in AZA patients (P<0.05). In association with early acute rejections (ARE), projected half-life was 14.4/4.5 years in patients with and 18.7/14.5 years without rejection in the MMF/AZA group, respectively. CONCLUSIONS: MMF based protocols improved long-term graft survival without an increase in side effects. Early ARE were associated with worse half-life of the graft, although more stressed in the AZA group. Thus, to improve quality of life in children for very long-term outcome, ARE should be further decreased and renal function should be better preserved. PMID- 17460561 TI - Elevated interleukin-4 expression in severe recurrent hepatitis C virus after liver transplantation. AB - BACKGROUND: Chronic hepatitis C virus (HCV) is usually associated with high levels of hepatic interleukin (IL)-2 and low levels of IL-4 transcripts. HCV frequently recurs after liver transplantation, and its course is accelerated in this setting. We compared in situ expression of IL-2 and IL-4 in transplanted and nontransplanted patients with HCV. METHODS: A total of 74 liver biopsy specimens were studied; 52 came from transplanted patients, 38 of whom were HCV-positive (17 mild and 21 severe cases of recurrent HCV) and 22 came from nontransplanted patients, 17 of whom were HCV-positive (7 mild and 10 severe cases of HCV). The expression of IL-2 and IL-4 mRNA and IL-4 protein was studied using the reverse transcriptase polymerase chain reaction and immunohistochemical methods, respectively. RESULTS: IL-2 transcript levels were significantly higher in severe than in mild HCV in both liver graft recipients and nontransplanted patients. However, IL-2 levels were higher in nontransplanted than in transplanted patients. IL-4 transcripts and protein were preferentially detected in graft recipients with severe recurrent HCV. CONCLUSION: IL-4 expression is elevated in severe recurrent HCV and may play a role in the progression of hepatic lesions after liver transplantation. PMID- 17460562 TI - Salvage chemotherapy for refractory and relapsed posttransplant lymphoproliferative disorders (PTLD) after treatment with single-agent rituximab. AB - BACKGROUND: Single-agent rituximab has demonstrated encouraging efficacy and tolerability in posttransplant lymphoproliferative disorders (PTLDs) failing to respond to immunosuppression reduction (IR). This retrospective analysis was undertaken to determine the efficacy and safety of salvage therapy in recipients of solid organ transplants with progression of PTLD after rituximab first-line therapy. METHODS: Eleven patients who had received IR and single-agent rituximab were analyzed. Of these, 10 had received CHOP salvage chemotherapy. One patient with limited disease received tumor irradiation and further IR. Most patients (73%) had late PTLD (median onset of disease 145 months posttransplant), and most (83%) had monomorphic histology; 36% had EBV-association. RESULTS: IR and irradiation therapy re-induced complete remission (CR) and allowed long-term disease control in a patient with polymorphic PTLD relapse. CHOP therapy achieved CR in five (50%) and partial remission (PR) in two (20%) patients. Four of five (80%) patients achieving CR remained in CR at a median follow-up of 44.2 months. Of the patients achieving PR, one is currently alive, and the second died from transplant rejection after converting to CR after consolidative chemotherapy. Patients with stable disease (two) and progressive disease (one) have died from PTLD. There was one possible CHOP-associated death (acute cardiac event) and two patients had to be switched to less-toxic monotherapies. Median overall survival was 46.5 months (95% confidence interval: 23.6-49.1 months). CONCLUSIONS: CHOP salvage therapy achieved a favorable overall response rate of 70% in this setting, indicating that PTLD generally remains chemotherapy-sensitive after progression following first-line rituximab. PMID- 17460563 TI - Laboratory test variability and model for end-stage liver disease score calculation: effect on liver allocation and proposal for adjustment. AB - BACKGROUND: The use of the Model for End-Stage Liver Disease (MELD) score to prioritize patients on liver waiting lists must take the bias of different laboratories into account. METHODS: We evaluated the outcome of 418 patients listed during 1 year whose MELD score was computed by two laboratories (lab 1 and lab 2). The two labs had different normality ranges for bilirubin (maximal normal value [Vmax]: 1.1 for lab 1 and 1.2 for lab 2) and creatinine (Vmax: 1.2 for lab 1 and 1.4 for lab 2). The outcome during the waiting time was evaluated by considering the liver transplantations and the dropouts, which included deaths on the list, tumor progression, and patients who were too sick. RESULTS: Although the clinical features of patients were similar between the two laboratories, 36 (13.1%) out of 275 were dropped from the list in lab 1, compared to 5 (3.5%) out of 143 in lab 2 (P<0.01). The differences were mainly due to the deaths on the list (8% lab 1 vs. 2.1% lab 2, P<0.05). The competing risk analysis confirmed the different risk of dropout between the two labs independently of the MELD score, blood group, and preoperative diagnosis. The bias on MELD calculation was considered and bilirubin and creatinine values were "normalized" to Vmax of lab 1 (corrected value=measured value x Vmax lab 1/Vmax lab 2). By comparing receiver operating characteristic curves, the ability of MELD to predict the 6-month dropouts significantly increased from an area under the curve of 0.703 to 0.716 after "normalization" (P<0.05). CONCLUSIONS: Normalization of MELD is a correct and good compromise to avoid systematic bias due to different laboratory methods. PMID- 17460564 TI - Effects of activin A on pancreatic ductal cells in streptozotocin-induced diabetic rats. AB - BACKGROUND: The shortage of islets for transplantation has led to find alternative insulin producing cells. Pancreatic progenitor cells in the duct have the potential to grow and differentiate into endocrine cells. In this study, we examined whether activin A can promote the expansion and/or differentiation of ductal cells into insulin-producing cells. METHODS: Pancreatic ductal cells were treated with activin A for differentiation into endocrine cells, and transplanted into the renal subcapsular space of streptozotocin (STZ)-induced diabetic rats. The identity of the endocrine cells was confirmed by immunostaining and analysis of the expression of transcription factors and endocrine genes by reverse transcriptase polymerase chain reaction. RESULTS: Activin A treatment significantly increased the DNA synthesis and the expression of insulin I, insulin II, PDX-1, Nkx 6.1, Glut-2, Pax-4, Pax-6, and Ngn-3. De novo synthesis of insulin in activin A-treated ductal cells was observed by the immunocytochemical detection of C-peptide and the differentiated ductal cells secreted significantly increased amount of insulin compared to nontreated ductal cells in response to glucose stimulation. When activin A-treated ductal cells were transplanted on STZ induced diabetic rats, blood glucose levels were normalized and the removal of the transplanted kidney resulted in return to hyperglycemia. CONCLUSIONS: The pancreatic ductal cells could be efficiently differentiated into insulin secreting cells by activin A treatment in vitro and normalize hyperglycemia in vivo. PMID- 17460565 TI - Pathways of helper CD4 T cell allorecognition in generating alloantibody and CD8 T cell alloimmunity. AB - BACKGROUND: The relative contributions of the "direct" and "indirect" pathways of CD4 T cell allorecognition in providing help for generating effective humoral and CD8 T cell alloimmunity remain unclear. Here, the generation of alloantibody and cytotoxic CD8 T cell responses to a vascularized allograft were examined in a murine adoptive-transfer model in which help could only be provided by transferred CD4 T cells recognizing alloantigen exclusively through the direct pathway. METHODS: Rejection kinetics and the development of alloantibody and cytotoxic CD8 T cell responses to MHC-mismatched H-2d heart grafts were compared when CD4 T cell help was present (wild-type H-2d recipients), or absent (CD4 T cell deficient, MHC class II-/- H-2b recipients [B6CII-/-]), or available only through the direct pathway (B6CII-/- mice reconstituted with wild-type CD4 T cells). RESULTS: BALB/c allografts were rejected by B6 mice rapidly (median survival time [MST] 7 days) with strong CD8 T cell effector and alloantibody responses, but were rejected by B6CII-/- mice more slowly (MST 23 days), with markedly reduced CD8 T cell responses and no detectable alloantibody. CD4 T cell reconstitution of B6CII-/- recipients accelerated heart graft rejection to near that of wild-type recipients (MST 13 days), with complete restoration of cytotoxic CD8 T cell responses but without detectable IgM or IgG alloantibody. CONCLUSIONS: Different pathways of helper T cell allorecognition are responsible for generating humoral and CD8 T cell alloimmunity. CD4 T cell help provided exclusively through the direct pathway generates strong cytotoxic CD8 T cell responses that effect rapid heart graft rejection. PMID- 17460566 TI - Influence of angiotensin II on expression of toll-like receptor 2 and maturation of dendritic cells in chronic cyclosporine nephropathy. AB - BACKGROUND: Angiotensin (Ang) II plays an important role in immune regulation. We evaluate the influence of the renin-angiotensin system (RAS) in the innate immune response caused by cyclosporine A (CsA)-induced renal injury. METHODS: Two separate studies were performed in Sprague Dawley rats. First, losartan (LSRT, 10 mg/kg per day) was concurrently administered with CsA (15 mg/kg per day) for 28 days. Second, AngII (435 ng/kg/min) was infused with or without LSRT for 14 days. RESULTS: AngII blockade with LSRT decreased toll-like receptor (TLR) 2 mRNA and protein expression, expression of tumor necrosis factor (TNF)-alpha mRNA, and expression of major histocompatibility complex class II antigen, which was upregulated in CsA-induced renal injury. The increased number of matured dendritic cells (DCs) in CsA-induced renal injury was also decreased by concomitant treatment of LSRT. Direct infusion of AngII increased TNF-alpha mRNA, TLR2 mRNA, and protein and the number of DCs, compared with the control rat kidney. In contrast, concomitant treatment of LSRT decreased all parameters. CONCLUSION: AngII plays a pivotal role in activating the innate immune response in CsA-induced renal injury. PMID- 17460567 TI - Inhibition of ERK1/2 activation by phenolic antioxidants protects kidney tubular cells during cold storage. AB - BACKGROUND: Cold storage of tissues induces reactive oxygen species (ROS), which contribute to cell injury. We have compared different antioxidants in protection of renal tubular cells against hypothermia injury and studied their effect on cold-induced mitogen-activated protein (MAP) kinase activation. METHODS: Cultured renal tubular epithelial cells (LLC-PK1) were stored in University of Wisconsin solution supplemented with compounds tested for 16 hr at 4 degrees C. Release of lactate dehydrogenase and cellular adenosine triphosphate were measured. Activation of MAP kinases was determined by Western blotting. Intracellular ROS were monitored with a fluorescent probe. RESULTS: Cold storage resulted in a substantial loss of cell viability. The simple phenol butylated hydroxyanisol (BHA) most effectively prevented hypothermia-induced cell injury, whereas about 100-fold higher concentration of the polyphenol epigallocatechin gallate (EGCG) was needed, although EGCG most effectively scavenged intracellular ROS elicited by serum withdrawal. The MEK inhibitor U0126 and reduced nicotinamide adenine dinucleotide phosphate (NADPH) oxidase inhibitor diphenyleneiodonium effectively protected the cells against hypothermia injury. ERK1/2 was rapidly activated during chilling of the cells and this was inhibited by BHA but not by EGCG. CONCLUSION: The results suggest that chilling of renal epithelial cells induces ROS generation by NADPH oxidase, which leads to rapid activation of the MEK ERK1/2 cascade and initiation of cell injury. This can be prevented by antioxidants. PMID- 17460568 TI - Addition of cyclophosphamide to T-cell depletion-based nonmyeloablative conditioning allows donor T-cell engraftment and clonal deletion of alloreactive host T-cells after bone marrow transplantation. AB - BACKGROUND: Bone marrow (BM) chimerism has been shown to have a beneficial effect on allograft survival. We recently found that production of donor T-cells was highly correlated with induction of tolerance in minimally conditioned chimeras. In the present studies, we demonstrate that nonmyeloablative conditioning and BM cell infusion modulate innate and adaptive host immune responses. METHODS: Chimeras were generated by bone marrow transplantation (B10.BR to B10). Recipients were preconditioned with T-cell depleting antibodies and total body irradiation with or without cyclophosphamide. Donor-specific tolerance was tested by skin grafting. RESULTS: Transfer of tolerant splenocytes to immunocompetent secondary recipients did not transfer tolerance, nor did infusion of tolerant CD4+/CD25+ T-cells into chimeras without donor T-cell production, demonstrating that linked suppression is an unlikely mechanism in tolerance induction in the context of BM cell infusion. The addition of a single dose of cyclophosphamide to the conditioning enhanced engraftment and tolerance. This was associated with production of donor T-cells and effective clonal deletion, and a significant reduction in activated recipient plasmacytoid dendritic cells (pDC) and natural killer (NK) cells. Chimeras without donor T-cell production that eventually lost their chimerism did not generate an antidonor humoral response, whereas unconditioned controls infused with similar numbers of BM cells did, indicating that infusion of donor BM cells into conditioned recipients induced immune deviation for adaptive B-cell immunity, preventing sensitization to major histocompatibility complex (MHC) alloantigens. CONCLUSIONS: These results demonstrate that recipient T-cells, pDC, and NK cells contribute to the host barrier for establishing chimerism, implicate deletional tolerance as the mechanism for total body irradiation-based nonmyeloablative conditioning for BM transplantation, and show a beneficial effect of BM cells in preventing sensitization to MHC alloantigens. PMID- 17460569 TI - Improved definition of human leukocyte antigen frequencies among minorities and applicability to estimates of transplant compatibility. AB - BACKGROUND: HLA population data can be applied to estimates of waiting time and probabilities of donor compatibility. Registry data were used for derivation of HLA antigen and haplotype frequencies in a 1996 report. At that time there were several instances of significant deviation from Hardy Weinberg Equilibrium (HWE). Because molecular typing has been increasingly used since 1996, analysis of recent donor phenotypes should provide more accurate HLA frequencies. METHODS: HLA frequencies were derived from the phenotypes of 12,061 donors entered into the Organ Procurement and Transplantation Network registry from January 1, 2003 to December 31, 2004. Frequencies for HLA-A;B;DR and HLA-A;B, DR, DQ haplotypes were derived from 11,509 and 10,590 donors, respectively. Frequencies of the allele groups encoding serologic antigens were obtained by gene counting and haplotype frequencies were estimated using the expectation maximization algorithm. Fit to HWE was evaluated by an exact test using Markov Chain Monte Carlo methods. RESULTS: There was clear evidence of improved definition of rarer HLA antigens and haplotypes, particularly among minorities. The reported frequencies of broad antigens decreased overall for HLA-A, B, and DR, with concomitant increases in split antigens. Allele group genotypes among the major ethnic groups were in HWE with the single exception of HLA-A locus alleles among Asians. Improved HLA definition also permitted the first report of DR;DQ and A;B;DR;DQ haplotypes among U.S. donors. CONCLUSIONS: The noted improvements in HLA definition and the overall lack of significant deviation from HWE indicate the accuracy of these HLA frequencies. These frequencies can therefore be applied for representative estimates of the U.S. donor population. PMID- 17460570 TI - Genes associated with progression and recurrence of hepatocellular carcinoma in hepatitis C patients waiting and undergoing liver transplantation: preliminary results. AB - BACKGROUND: Liver transplantation (LT) represents a curative treatment for small hepatocellular carcinoma (HCC). Potentially curable higher-stage HCC patients are denied LT due to the lack of cancer markers that predict progression and recurrence. METHODS: Thirty-eight candidates for LT with hepatitis C virus (HCV) cirrhosis and HCC were studied. Gene expression (Gexp) analysis of tumor samples was performed using microarrays. RESULTS: Twenty patients underwent transplantation, 13 progressed while waiting for transplantation, 4 are alive awaiting transplantation, and 1 died without progression while waiting for LT. Differences in GExp among patients who underwent LT or did not progress (n=25) versus those whose disease progressed while waiting for LT (n=13) were assessed. Thus, 54 probe sets (Pset) were significantly differentially expressed. Among LT patients, 10 Psets were differentially expressed between LT patients with the same explanted stage that recurred (n=5) versus LT patients who did not recur (n=5). Ninety-eight Psets were significantly associated with survival at the alpha=0.005 level. CONCLUSIONS: Here, we have identified genes associated with HCC progression in HCV-HCC patients awaiting LT transplantation. A limited number of genes were related to overall survival and cancer-free survival after LT. Incorporation of these molecular markers could help to improve organ allocation for HCV-HCC patients. PMID- 17460571 TI - HLA-specific B cells: I. A method for their detection, quantification, and isolation using HLA tetramers. AB - BACKGROUND: The development of highly sensitive and specific assays for detecting and characterizing HLA-specific antibodies has contributed to an appreciation of the extensive involvement of those antibodies in graft injury and dysfunction. However, understanding the regulatory processes of the humoral response to transplantation and the mechanisms underlying therapeutic agents and protocols for preventing and treating sensitization requires a way to study HLA-specific B cells. METHODS: Lymphocyte preparations enriched for B cells were stained with one or more of three different HLA tetramers. Tetramer-positive (tet+) B cells were enumerated and evaluated for an association of their frequencies with known sensitization. In some cases, tet+ B cells were isolated and placed in culture with supplements known to activate B cells in a nonspecific fashion. RESULTS: For all tetramers used, the frequencies of tet+ B cells were significantly higher (4.1%-5.5%) among sensitized patients than among nonsensitized patients (1.6% 3.2%, P<0.001). Binding of the tetramers occurred by the surface immunoglobulin antigen receptor with little or no binding to antibody captured in the Fc receptor. Cultured tet+ B cells produced antibodies specific for epitopes of the tetramer antigen. There appeared to be a certain amount of crossreactivity in the binding of tetramers. The frequency of CD27+ cells among tet+ B cells was higher, on average (34.4%-38.8%) than among all B cells (26.2%) whereas the frequencies of CD38 were comparable in the two groups. CONCLUSIONS: Staining with HLA tetramers provides a means for identifying, quantifying, and isolating HLA specific B cells. PMID- 17460572 TI - HLA-specific B cells: II. Application to transplantation. AB - BACKGROUND: Differences in the antibody response to allogeneic transplantation exist between groups defined by race or gender. These differences may reflect differences in immune competency and/or exposure to alloantigens. We have investigated the frequencies and phenotypes of HLA-specific B cells to address those possibilities. METHODS: HLA-specific B cells were identified by staining with HLA tetramers (tet) as described previously and the distribution of CD27 and CD38 among those cells were measured in groups defined by various parameters. Possible correlation between frequencies of HLA-specific B cells and production of HLA-specific antibody after transplantation was also investigated. RESULTS: We found no correlation between the frequencies of CD27+tet+ (33%-44% vs. 34%-36%) or CD38+tet+ (57%-65% vs. 59%-66%) B cells and a previous mismatch for the HLA antigen of the tetramer. However, there was an increase in CD38+tet+ B cells among patients making antibody to the tetramer antigen (67%-72% vs. 53%-56%). Blacks had lower frequencies of CD27+ B cells than did whites (11.8% vs. 28.9%, P=0.003), but had greater increases of these cells among tet+ cells than did whites. There was a higher frequency of tet+ B cells among patients who developed "new" antibody to the HLA antigen (3.9%-8.6%) of the tetramer after transplantation than among those who did not (1.1%-3.7%). CONCLUSIONS: The phenotype of HLA-specific B cells reflects current or historic sensitization to HLA and may reflect inherent differences between groups defined by race and/or gender. The frequencies of HLA-specific B cells may predict patients at risk for production of donor-specific antibody after transplantation. PMID- 17460573 TI - Early urethral (Foley) catheter removal positively affects length of stay after renal transplantation. PMID- 17460574 TI - N,N,N-trimethylglycine (betaine) improves analysis of CDR3 diversification in children reconstituting their immune repertoire after hematopoietic stem-cell transplantation. PMID- 17460575 TI - Tacrolimus dose in black renal transplant recipients. PMID- 17460576 TI - Response: organ donation and the law. PMID- 17460577 TI - Long-term patient satisfaction after revision rhinoplasty. AB - OBJECTIVE: In cases of dissatisfaction after rhinoplasty for esthetic or functional reasons, or both, revision rhinoplasty may be advocated to improve the patients' nasal performance. In contrast to studies on primary rhinoplasty, no objective outcome evaluation parameter has been validated in revision rhinoplasty, nor has there been a study specifically looking at long-term satisfaction after revision rhinoplasty. We aim at studying patient satisfaction after a long-term follow-up of revision rhinoplasty performed at an academic referral center. METHODS: At a mean period of 2.5 years after revision rhinoplasty, 90 patients who underwent a revision rhinoplasty at the Academic Medical Center in Amsterdam, The Netherlands, were sent a questionnaire on general satisfaction together with the validated rhinoplasty outcome evaluation (ROE) questionnaire. RESULTS: Of the 46 patients who responded to the questionnaire, 88% experienced a significant improvement in nasal performance by revision rhinoplasty, and 79% would choose to undergo the revision procedure again. Subgroup analysis of the ROE scores demonstrated that good satisfaction was mainly obtained in the young adult and middle-aged groups, without major differences between males and females. Satisfaction was unrelated to the open or closed technique used for revision, or to the graft material used. Interestingly, an inverse correlation was found between satisfaction and the number of previous rhinoplasties. CONCLUSIONS: This retrospective study demonstrates that revision rhinoplasty in an academic practice provides most patients with long-standing satisfaction. PMID- 17460578 TI - Clinical significance of olfactory event-related potentials related to orthonasal and retronasal olfactory testing. AB - OBJECTIVE: The purpose of this study was to evaluate the likelihood of recording olfactory event-related potentials (OERPs) in patients with an olfactory dysfunction and to correlate the electrophysiological responses to orthonasal and retronasal olfactory testing. DESIGN/MATERIALS AND METHODS: This was a prospective study of 65 patients with different origins of their olfactory loss. Orthonasal olfactory function was assessed with the "Sniffin' Sticks" test (orthonasal score; maximal score 48) and retronasal olfactory function with odorized powders presented intraorally (retronasal score; maximal score 20). The OERPs were obtained after presentation of 2-phenyl ethyl alcohol, the selected olfactory stimulus. Causes of olfactory dysfunction included postinfectious olfactory loss (n = 15), head trauma (n = 26), nasal polyposis (n = 15), and mixed causes (idiopathic, toxic, drug induced) (n = 9). RESULTS: Based on orthonasal testing, 32 and 33 patients were diagnosed with anosmia and hyposmia, respectively. Twenty-two patients from the hyposmic group demonstrated reliable OERPs. No OERPs were recorded in the anosmic group. Prevalence of OERPs in a cohort of patients with olfactory dysfunction was 33.8% (22 of 65). Median score (expressed as the percentage of the maximal score that could be obtained theoretically) in which OERPs were recorded was 50% (24 of 48) with orthonasal testing and 80% (16 of 20) with retronasal testing. CONCLUSIONS: Patients with olfactory dysfunction usually demonstrate OERPs in one third of the cases. When olfactory dysfunction is in the range that separates normosmic subjects from anosmic patients, patients may have identifiable OERPs. Interpretation of both orthonasal and retronasal psychophysical olfactory testing should be supported by the recording of OERPs in a clinical setting. PMID- 17460579 TI - Bone regeneration of canine skull using bone marrow-derived stromal cells and beta-tricalcium phosphate. AB - OBJECTIVE: The aim of this study was to regenerate high-quality cranial bone using tissue engineering techniques, with subsequent extension to clinical application. Our previous study with a 3-month observation period indicated that a composite scaffold composed primarily of beta-tricalcium phosphate (TCP) had the potential for cranial bone regeneration. In this study, we investigated whether bone marrow derived stromal cells (BSCs) could promote the regeneration of cranial bone as determined after 3 and 6 months. STUDY DESIGN: The pilot study was conducted with 14 adult beagle dogs. MATERIALS AND METHODS: Craniotomy was performed in the same manner used clinically. The bone defect (2 x 2 cm) was created at each canine temporoparietal region. The test animals were divided into three groups. In group I, the bone defect was closed by replacing the original free bone flap without filling the residual gaps. In group II, the gap was filled with a composite scaffold consisting of collagen coated beta-TCP and autologous bone fragments with fibrin glue. In group III, autologous cultured BSCs and the composite scaffold were used to fill the gap. The sites of craniotomy were analyzed with three-dimensional computed tomography and histologic examination 3 and 6 months after the operation. RESULTS: Bone regeneration was observed in groups II and III, with more extensive formation in group III than in group II. In group I, bone regeneration was not observed. CONCLUSION: This study showed that BSCs have the potential to promote cranial bone regeneration and confirmed the efficacy of a composite scaffold made of beta-TCP and autologous bone fragments with fibrin glue. PMID- 17460580 TI - Inverse association between T-cell immunoglobulin and mucin domain-1 and T-bet in a mouse model of allergic rhinitis. AB - BACKGROUND: It has been suggested that human hepatitis A virus cellular receptor, also known as T-cell immunoglobulin and mucin domain-1 (TIM-1), plays an important role in the development of allergic diseases on the basis of epidemiologic data, but the molecular mechanism has been unclear. In a murine model of ovalbumin (OVA)-sensitized allergic rhinitis (AR), we examined the expression of TIM-1 and its correlation with T helper1-associated transcription factor, T-bet, as a potential mediator of T-cell immunoglobulin expression. METHODS: Mice were challenged intranasally with OVA to elicit AR. The expression of TIM-1 in nasal tissues was examined by real-time reverse-transcription polymerase chain reaction (RT-PCR), and the surface expression of TIM-1 in peripheral blood mononuclear cells was evaluated by means of flow cytometry. In addition, the expression of TIM-1 as well as T-bet in splenic lymphocytes was examined by Western blotting. RESULTS: TIM-1 mRNA was increased significantly in nasal tissues (P < .05) as seen by real-time RT-PCR. Flow cytometry indicated a differential TIM-1 expression of 135.5 +/- 34.2 in the AR group versus 51.1 +/- 10.9 in the control group (P < .05). The mean values of normalized TIM-1 were 0.43 +/- 0.18 and 0.21 +/- 0.10 in AR and control groups, respectively, whereas the mean values of normalized T-bet were 0.22 +/- 0.13 and 0.67 +/- 0.17 in the AR and control groups, respectively. There was a significant difference in the production of TIM-1 as well as T-bet in AR mice versus control mice (P < .05). The increased production of TIM-1 correlated significantly with the decreased T bet in spleen tissue of AR mice (r = -0.52, P < .05). CONCLUSION: Our experimental model recapitulates an increase in lymphocyte TIM-1 expression seen in AR both locally and systemically. Our results also demonstrate an inverse relationship between lymphocyte TIM-1 and T-bet expression, suggesting a possible mechanism that TIM-1 influences the development of AR. PMID- 17460581 TI - Role of genetic factors and inflammation in age-related macular degeneration. AB - Complement factor H (CFH) has been implicated in the predisposition to advanced forms of age-related macular degeneration (AMD). The purpose of this review is to highlight recent discoveries implicating single nucleotide polymorphisms on 1q32, 6p21, and 10q26 in the risk for development of AMD. In addition, the central role of CFH in the complement cascade and its role in the inflammatory hypothesis for AMD are reviewed. PMID- 17460582 TI - Fundus changes in central retinal artery occlusion. AB - PURPOSE: To investigate systematically the ophthalmoscopic fundus findings associated with central retinal artery occlusion (CRAO). METHODS: The study included 240 consecutive patients (248 eyes) with CRAO. The eyes underwent detailed fundus evaluation and stereoscopic color fundus photography at initial and follow-up visits. Patients without evidence of giant cell arteritis were advised to have carotid Doppler imaging and echocardiography to determine the source of emboli. CRAO was classified into 3 types: permanent CRAO (175 eyes), permanent CRAO with cilioretinal artery sparing (35 eyes), and transient CRAO (38 eyes). In the three types of CRAO, acute-phase and late-phase changes in the retina, optic disk, and retinal vessels were evaluated. RESULTS: The main findings during the initial examination in our clinic for permanent CRAO were retinal opacity in the posterior pole (58%), cherry-red spot (90%), box-carring (19%), retinal arterial attenuation (32%), and optic disk edema (22%) and pallor (39%). The most frequent findings identified at the late stage, based on survivorship curves, were optic atrophy (91%), retinal arterial attenuation (58%), cilioretinal collaterals (18%), and macular retinal pigment epithelial changes (11%). Compared with permanent CRAO, permanent CRAO with cilioretinal artery sparing was associated with a lower incidence of all macular and optic disk abnormalities. For transient CRAO, the incidence of initial findings varied greatly compared with the other types. Intraarterial emboli were observed in 20% of patients. Carotid Doppler evaluation identified carotid vascular plaques in 67% of patients tested and hemodynamically significant (>50%) obstruction in 32%. Four percent of CRAOs presented with simultaneous bilateral onset. CONCLUSIONS: The type and incidence of fundus findings at the initial visit and in the late phase of CRAO vary by its type. This study confirms that retinal opacity is predominantly evident in the posterior retina, that optic disk findings at presentation are common, that CRAO associated with normal-appearing retinal vessels and/or optic disk is not rare, and that observation of emboli is infrequent. Clinicians should be aware of the various presentation findings during the acute and late stages of CRAO and its various types. A complete picture of CRAO is provided by combined information of our clinical and experimental studies of CRAO. PMID- 17460583 TI - Anticoagulation with warfarin in vitreoretinal surgery. AB - PURPOSE: To describe the clinical course of patients undergoing vitreoretinal procedures while receiving systemic anticoagulation with warfarin. METHODS: We reviewed patient demographics, ocular findings, and clinical courses for 25 patients receiving systemic anticoagulation with warfarin who subsequently underwent vitreoretinal surgery. RESULTS: Patient ages ranged from 49 years to 81 years (median, 69 years). Indications for anticoagulation included atrial fibrillation, cerebrovascular disease, deep vein thrombosis, prosthetic heart valves, and hypercoagulable state. Follow-up ranged from 4 months to 36 months (median, 19.5 months). The international normalized ratio ranged from 1.5 to 3.1 (median, 2.0). Final vision after surgery ranged from 20/20 to 20/400 (median, 20/100). One patient who underwent scleral buckling and external drainage of subretinal fluid had an intraoperative subretinal hemorrhage associated with the drainage procedure. In all other patients, no intraoperative complications occurred. CONCLUSION: Cessation of therapy with warfarin may not be necessary in patients receiving anticoagulation who are undergoing vitreoretinal procedures. Successful visual and anatomical results may be achieved after vitreoretinal surgery for patients receiving anticoagulation with warfarin. The management of anticoagulation should occur in conjunction with the patient's internist to allow a clear understanding of the potential systemic risks of cessation of warfarin treatment preoperatively. PMID- 17460584 TI - Outcomes of transconjunctival sutureless 25-gauge vitrectomy with silicone oil infusion. AB - PURPOSE: To evaluate the outcomes and complications of surgical management with 25-gauge pars plana vitrectomy (PPV) and silicone oil (SO) tamponade in complex vitreoretinal diseases. METHODS: Retrospective review of a consecutive, interventional case series at a single center. RESULTS: Thirty-five eyes of 35 patients were included in the study. The indications for vitrectomy included tractional retinal detachment (11 eyes), macular hole (6 eyes), proliferative vitreoretinopathy or recurrent retinal detachment (9 eyes), neovascular glaucoma (3 eyes), giant retinal tear (3 eyes), and pathologic myopia with epiretinal membrane or macular hole (3 eyes). All patients underwent 25-gauge PPV with either 1,000-centistoke (n = 31) or 5,000-centistoke (n = 4) SO tamponade infused through a 24-gauge angiocatheter. No intraoperative complications were noted. The median preoperative visual acuity was counting fingers (range, 20/50 to light perception). The median postoperative visual acuity after a median follow-up of 6 months (range, 1-19 months) was 20/200 (range, 20/30 to light perception). A small subconjunctival SO bleb was identified in two patients. Recurrent retinal detachment occurred in three patients. No significant complications relating to the use of SO in the setting of 25-gauge PPV occurred. CONCLUSIONS: Advances in 25-gauge PPV instrumentation have enabled expanding indications for 25-gauge PPV. 25-Gauge PPV with SO tamponade is safe and efficient and can be considered in the surgical management of complex vitreoretinal disease. PMID- 17460585 TI - Primary silicone oil tamponade in the management of severe intraocular foreign body injuries: an 8-year follow-up. AB - PURPOSE: To evaluate the long-term outcome of pars plana vitrectomy and primary silicone oil tamponade in patients with severe intraocular foreign body (IOFB) injuries and high risk of proliferative vitreoretinopathy (PVR). METHODS: This retrospective consecutive study included 23 patients with severe IOFB injuries who had extensive lacerations including sclera, choroid, and retina, and were complicated by predictive factors for elevated proliferative activity and an unfavorable outcome. All patients underwent pars plana vitrectomy, removal of the IOFB, and primary silicone oil tamponade and were followed up for a mean 8.9 years. Main functional outcome was assessed as final best-corrected visual acuity. Anatomic success was defined as permanent retinal attachment. RESULTS: PVR occurred in 70% of all eyes and required 16 revisions. Silicone oil was removed in 78% of the eyes after a mean tamponade duration of 9.1 months. Complete retinal attachment was achieved in 83% of the eyes. Three eyes developed a persisting hypotony that was stabilized under permanent silicone oil. Functional stabilization was observed in the third year resulting in a final visual acuity of 20/630. Useful vision of better than 20/400 could be preserved in 55% of the patients. Only one eye underwent a late enucleation after 6.8 years. CONCLUSIONS: Primary silicone oil stabilizes the retina during the critical period of active PVR and may limit the visual loss in selected high-risk eyes in the long term. PMID- 17460586 TI - Assessment of macular function by focal electroretinogram and pattern electroretinogram before and after epimacular membrane surgery. AB - PURPOSE: To evaluate macular function before and after surgical peeling of idiopathic epimacular membrane (EMM). METHODS: Logarithm of the minimal angle of resolution visual acuity and results of focal (central 9 x 9 degrees) electroretinogram (fERG), pattern electroretinogram (pERG), and optical coherence tomography (OCT) assessment of macular volume were evaluated for 22 eyes of 22 patients (mean age +/- SD, 63.20 +/- 10.0 years) with EMM preoperatively (baseline) and 6 months after surgical peeling. Preoperative visual acuity and fERG and pERG amplitudes observed in EMM eyes were compared with those in 15 age matched control eyes. RESULTS: In the preoperative evaluation, EMM eyes had a significant (P < 0.01; one-way analysis of variance) reduction in visual acuity and fERG and pERG amplitudes and an increase in OCT macular volume when compared with controls. In EMM eyes, the decrease in visual acuity was significantly correlated (P < 0.01, Pearson test) to the reduction in fERG and pERG amplitudes. At the postoperative evaluation, EMM eyes had a correlated significant (P < 0.01) increase in visual acuity, fERG amplitude, and pERG amplitude with respect to the preoperative values. All EMM eyes had a significant (P <0.01) reduction in macular volume, and retinal microanatomy was restored to normal conditions. CONCLUSION: In EMM eyes, the decrease in visual acuity is related to dysfunction of both preganglionic (abnormal fERG) and ganglionic (abnormal pERG) macular elements. Surgical removal of EMM may induce improvement of the function of both outer and innermost macular retinal layers, leading to a related increase in visual acuity. PMID- 17460587 TI - Management of submacular hemorrhage with intravitreal injection of tissue plasminogen activator and expansile gas. AB - PURPOSE: To evaluate the clinical outcome of intravitreal tissue plasminogen activator (tPA) and expansile gas injection as a minimally invasive treatment for submacular hemorrhage (SMH). METHODS: This study was a retrospective clinical case series examining 104 eyes that received an intravitreal injection of 30-100 mcg of tPA and expansile gas (SF6 or C3F8) for SMH. The main outcomes evaluated were visual acuities (VA), anatomic displacement of submacular blood, and surgical complications. RESULTS: : A total of 85, 77, and 81 eyes were available at 1 week, 3 months, and 12 months follow up, respectively. Postoperatively, > or = 2 Snellen lines improvement were achieved in 43/85 eyes (51%) at 1 week, 49/77 eyes (63%) at 3 months, and 52/81 eyes (64%) at 12 months. Postoperative VA improvement was significantly associated with preoperative VA, submacular blood displacement, and the underlying cause of SMH. Diagnostic postoperative angiogram and clinical examination were possible at 8.2 +/- 7.4 weeks and 9.5 +/- 7.4 weeks, respectively. The observed complications included breakthrough vitreous hemorrhage in 8 eyes (8%) and retinal detachment in 3 eyes (3%). CONCLUSIONS: In this retrospective series, intravitreal injection of tPA and expansile gas was shown to be a safe and effective technique that can improve VA in most eyes with SMH and assist in the diagnosis of the underlying cause. PMID- 17460588 TI - Pneumatic displacement of submacular hemorrhage: safety, efficacy, and patient selection. AB - PURPOSE: To investigate the safety and efficacy of pneumatic displacement of submacular hemorrhage without the use of intravitreal tissue plasminogen activator (tPA). METHODS: In a prospective, consecutive, single-center, noncomparative, interventional case series, 20 consecutive patients with submacular hemorrhage due to various etiologies (duration, 1-30 days; visual acuity, hand movements to 20/125) were treated with intravitreal perfluoropropane gas injection followed by prone positioning for 5 days to 7 days. Patients were observed 1 day, 7 days, 30 days, 3 months, 6 months, and 1 year after the procedure. RESULTS: Submacular blood was completely or partially displaced from the fovea in 16 of 20 eyes within 7 days after the injection. Mean best-corrected visual acuity improved from 1.6 to 0.72 logarithm of minimum angle of resolution, which was statistically significant (Wilcoxon signed rank test, P = 0.001). Final visual acuity was 20/63 or better in 10 eyes (50%). The change in proportion of eyes with best-corrected visual acuity of 20/63 or better after treatment was statistically significant (McNemar test, P = 0.002). Four patients developed nonresolving vitreous hemorrhage, which necessitated vitrectomy. CONCLUSION: The results of pneumatic displacement of submacular hemorrhage using perfluoropropane gas are encouraging, thus questioning the role of tPA in the management of such cases. PMID- 17460589 TI - Hemorrhagic complications after photodynamic therapy for polypoidal choroidal vasculopathy. AB - PURPOSE: To evaluate the clinical features and risk factors of hemorrhagic complications in eyes with polypoidal choroidal vasculopathy (PCV) after photodynamic therapy (PDT). METHODS: We retrospectively reviewed data for 91 consecutive eyes of 85 patients who underwent PDT for the treatment of PCV. The diagnosis of PCV was based on indocyanine green angiographic findings, showing a branching vascular network terminating in polypoidal swelling. The greatest linear dimension included all polypoidal lesions, leaking vascular network, and type 2 choroidal neovascularization. RESULTS: During the follow-up period after PDT, postoperative subretinal hemorrhage was seen in 28 (30.8%) of 91 eyes. In 22 (78.6%) of these 28 eyes, subretinal hemorrhage was absorbed without treatment. In 6 eyes (21.4%), however, bleeding resulted in vitreous hemorrhage, and 2 eyes underwent pars plana vitrectomy. Although visual acuity was maintained or increased in 18 (81.8%) of 22 eyes with subretinal hemorrhage alone, it decreased significantly in 3 (50.0%) of 6 eyes with postoperative vitreous hemorrhage. Various systemic diseases and medication with an anticoagulant had no correlation with these hemorrhagic complications. Laser irradiation spot size for PDT was significantly larger in eyes with postoperative vitreous hemorrhage (P = 0.017) than in those without. CONCLUSION: Subretinal hemorrhage after PDT can be a common complication in patients with PCV and may have a minor effect on visual outcome. However, postoperative hemorrhage is occasionally so massive that it leads to vitreous hemorrhage and poor visual prognosis. When considering PDT for eyes with a large PCV lesion, ophthalmologists should be aware of the risk of serious hemorrhagic complications. PMID- 17460590 TI - Photodynamic therapy with verteporfin for choroidal neovascularization associated with retinal pigment epithelial detachment in age-related macular degeneration. AB - PURPOSE: To assess the effectiveness of photodynamic therapy (PDT) with verteporfin for choroidal neovascularization (CNV) associated with retinal pigment epithelium detachment (PED) in age-related macular degeneration. METHODS: Thirty eyes of 26 patients with CNV and PED were treated with PDT. The eyes were divided in two groups based on CNV location in relation to PED; group 1 included 13 eyes with CNV within PED, and group 2 included 17 eyes with CNV at the edge of PED. The median follow-up was 16 months. RESULTS: Patients received a mean +/- SD of 2.83 +/- 1.26 treatments (range, 1-6 treatments). In the whole cohort, the mean preoperative visual acuity changed from 20/144 (0.86 +/- 0.42 logarithm of minimal angle of resolution [logMAR]) to 20/182 (0.96 +/- 0.51 logMAR; P = 0.39) at month 18. Five eyes (16%) gained a mean of 1.5 Snellen lines from baseline. Twelve eyes (40%) lost a mean of 1.7 Snellen lines of visual acuity. Vision in 13 eyes (44%) remained stable. In group 1, the mean visual acuity at month 12 was 20/303 (1.18 +/- 0.51 logMAR) and significantly (P = 0.015) worse than that, 20/110 (0.74 +/- 0.42 logMAR), in group 2. CONCLUSION: PDT can improve or stabilize visual function in 60% of eyes with vascularized PED. CNV at the edge of PED appears to respond more favorably to PDT. Appropriate patient selection and prompt treatment are essential to obtain the best outcomes after verteporfin therapy. PMID- 17460591 TI - Retinal thickness and visual thresholds measured in patients with retinitis pigmentosa. AB - PURPOSE: : To determine the association between retinal thickness and visual function in patients with retinitis pigmentosa (RP). METHODS: : Retinal thickness was estimated from optical coherence tomography (OCT) images obtained for six patients with RP. The thickness measurements were compared with dark-adapted rod and minimally light-adapted cone thresholds obtained by psychophysical testing using a Tubinger perimeter and with standard light-adapted Humphrey visual field (HVF) perimetric thresholds. RESULTS: : Four patterns of association between retinal thickness and visual function were observed: normal retinal thickness and normal visual thresholds; normal retinal thickness and normal cone thresholds but elevated rod thresholds; reduced retinal thickness and elevated rod and cone thresholds; and normal retinal thickness and normal Humphrey thresholds but elevated rod and cone thresholds by Tubinger perimetry. CONCLUSION: : Retinal thinning was observed only when both rod and cone thresholds were elevated. However, normal retinal thickness was not necessarily accompanied by normal visual sensitivity. The determination of retinal thickness by OCT and its association with psychophysical measurements of visual function could be useful for identifying those RP patients who might respond most optimally to therapeutic interventions. PMID- 17460592 TI - Macular function of successfully repaired macula-off retinal detachments. AB - PURPOSE: To assess the macular function of successfully repaired macula-off retinal detachments in the long term and to evaluate the relationship between duration of macular detachment and functional recovery. METHODS: Of 214 consecutive patients, 29 who were operated on because of unilateral retinal detachment with at least 6 months of follow-up and whose best-corrected visual acuity was better than 0.05 Snellen chart value in both eyes were examined. Operated eyes were included in group I, and the fellow eyes were included in group II. Macular functions were evaluated in terms of visual acuity, contrast sensitivity, color vision, visual field, and visual evoked potentials after reattachment. RESULTS: All macular functions except P100 amplitudes in the operated eyes were significantly less than those in the fellow eyes (P = 0.28) following reattachment after nearly 5 years. It was found that the longer the duration of macular detachment (except detachment duration of < or = 7 days), the smaller the increase in visual acuity, contrast sensitivity, and color vision defect scores (P = 0.000, P = 0.034, and P = 0.0003, respectively). CONCLUSIONS: Macular functions except P100 amplitude cannot be recovered completely 5 years after retinal reattachment. There was no relation between the duration of macular detachment and the mean deviation in visual field and the P100 latency and P100 amplitude difference between both eyes. PMID- 17460593 TI - Macular hole repair with limited nonsupine positioning. AB - PURPOSE: To assess the surgical success rates of modern macular hole repair with elimination of face down positioning. METHODS: A review of data for 72 eyes (from 102 consecutive cases) with idiopathic macular holes treated surgically between 1998 and 2004 was performed. Exclusion criteria consisted of macular hole for >1 year or of unknown duration and macular holes from secondary causes. All patients were evaluated and surgically managed by one surgeon (R.T.). RESULTS: Average preoperative best spectacle corrected visual acuity (BSCVA) was 20/170 (6/51). Six patients had a grade II hole, 60 patients had a grade III hole, and 6 patients, had a grade IV hole. Anatomical success was achieved in 92% of cases with 1 operation, and the average postoperative BSCVA was 20/46 (6/14). Six patients required additional surgical management to achieve anatomical success with an average postoperative BSCVA of 20/55 (6/16.5). The postoperative BSCVA improved an average of 5.7 lines from baseline. CONCLUSION: Favorable anatomical and BSCVA outcomes were achieved with the elimination of face down positioning in the postoperative period. Additional benefits are an increase in patient acceptance and compliance and the number of patients eligible for the procedure. PMID- 17460594 TI - Analysis of forces acting upon submacular hemorrhage in pneumatic displacement. AB - PURPOSE: To analyze the forces affecting pneumatic displacement of submacular hemorrhage and determine an efficient postoperative positioning method for patients. METHOD: Geometric analysis of the forces that act upon submacular hemorrhage in the absence and presence of a gas bubble. RESULTS: A component of gravity force of the hemorrhage parallel to the subretinal space is responsible for pneumatic displacement. This component is largest when the patient is looking straightforward and is absent when the patient is positioned facedown. CONCLUSION: Face down positioning is not optimal for pneumatic displacement of submacular hemorrhage and should be avoided. PMID- 17460595 TI - Indocyanine green and trypan blue: intracellular uptake and extracellular binding by human retinal pigment epithelial cells. AB - PURPOSE: To examine the intracellular uptake and extracellular binding of indocyanine green (ICG) and trypan blue (TB) by human retinal pigment epithelium (RPE) cells in vitro at clinically relevant concentrations. METHODS: Human RPE cells (ARPE19 cells) were exposed to 0.01%, 0.05%, and 0.25% ICG or 0.06%, 0.15%, and 0.25% TB for 15 seconds, 30 seconds, and 5 minutes, and the dye concentrations taken up by the cells were evaluated by spectrophotometric analysis. RESULTS: ICG was significantly taken up by ARPE19 cells after exposure to 0.05% for 5 minutes, 0.01% for 30 seconds, and 0.25% ICG for 15 seconds, whereas TB was not taken up by RPE cells under the conditions in this study. CONCLUSION: ICG is bound and taken up by RPE cells at concentrations lower than clinically used, and TB is not. Understanding the effects of dyes on ocular tissues leads to more effective and safer dye-assisted vitrectomy protocols, although the experimental setting is different from the clinical situation. PMID- 17460596 TI - Cutaneous malignant melanoma metastatic to the vitreous. AB - We describe a patient with dysplastic nevus syndrome and cutaneous melanoma in whom vitreous involvement was the first sign of metastatic disease. Cytologic examination of vitreous aspirate can usually confirm the diagnosis. Enucleation may be a treatment option if the vitreous is the sole metastatic site. PMID- 17460597 TI - Three-port 20-gauge transconjunctival pars plana vitrectomy. PMID- 17460598 TI - Diagnostic and therapeutic challenges. PMID- 17460599 TI - Cavernous hemangioma of the optic disk. PMID- 17460600 TI - Possibility of flap displacement during retinal surgery. PMID- 17460603 TI - Pulmonary endarterectomy in chronic thromboembolic pulmonary hypertension: short term functional assessment in a longitudinal study. AB - AIM: Presently, the surgical treatment choice in chronic thromboembolic pulmonary hypertension (CTEPH) consists in a pulmonary endarterectomy (PEA). The aim of the present study is the functional assessment of patients submitted to PEA both preoperatively and shortly after the intervention. A longitudinal study was developed to study the quality and quantity of functional performance possible in these subjects. METHODS: Twenty-two subjects were assessed immediately prior to PEA and 3 months later in order to obtain quantitative measurements of short-term functional recovery. The functional assessment included the 6-min walk test (6mWT), the measurement of the oxygen percent saturation (HbS%O(2)) and the degree of dyspnea subjectively perceived by each patient. RESULTS: Three months after the surgical intervention, there was a definite increase in the number of meters walked during the 6mWT with respect to preintervention; the difference between the distances walked in the 6mWT (6mWD) in the pre and post-PEA was statistically significant (Paired t-test P<0.001). CONCLUSION: In this study the 6mWT resulted to be a useful tool in the functional evaluation of patients affected by CTEPH and submitted to PEA. The average 6mWD significantly improved already at 3 months after the intervention, thus reaching the minimum limit of the range predicted for the healthy control, but remains lower than the average theoretical value predicted (about 75% of the same). PMID- 17460605 TI - Symmetric division of cancer stem cells--a key mechanism in tumor growth that should be targeted in future therapeutic approaches. AB - As cancer stem cells (SCs) drive tumor growth, it is only through the elimination of those cancer SCs that a pharmacologic cure can be attained. To study ways to develop drugs that target cancer SC, we investigated changes in cellular mechanisms and kinetics that occur in SC populations during colorectal cancer (CRC) development. We used computer modeling to determine which changes could give rise to exponential increases in both SC and non-SC populations in CRC. Our results show that the only mechanism that can explain how these subpopulations increase exponentially in CRC development involves an increase in symmetric SC cell division. This finding suggests that any systemic therapies designed to effectively treat CRC and other cancers must act to control or eliminate symmetrical cancer SC division in tumors, while minimally affecting normal SC division in non-tumor tissues. PMID- 17460606 TI - Factors affecting drug concentrations and QT interval during thioridazine therapy. AB - The objective of this study was to investigate factors affecting steady-state plasma concentrations of thioridazine. A cross-sectional study of patients receiving chronic thioridazine was employed. Common allelic variants of CYP2D6 and CYP2C19, as well as thioridazine and metabolite concentrations and QTc intervals, were determined. In 97 patients, dose-corrected plasma concentrations (C/Ds) of thioridazine and metabolites were correlated with age but not sex or CYP2C19 genotype. Patients with no functional CYP2D6 alleles (n=9) had significantly higher C/D for thioridazine (P=0.017) and the ring sulfoxide metabolite and a significantly higher thioridazine/mesoridazine ratio compared with those with >/=1 functional CYP2D6 allele (n=82). Smokers had significantly lower C/D for thioridazine, mesoridazine, and sulforidazine and significantly lower thioridazine/ring sulfoxide ratios than non-smokers. QTc interval was not significantly affected by CYP2D6 or CYP2C19 genotypes. Plasma concentrations of thioridazine are influenced by age, smoking, and CYP2D6 genotype, but CYP2D6 genotype does not appear to influence on-treatment QTc interval. PMID- 17460607 TI - SLCO1B1 (OATP1B1, an uptake transporter) and ABCG2 (BCRP, an efflux transporter) variant alleles and pharmacokinetics of pitavastatin in healthy volunteers. AB - To investigate the contribution of genetic polymorphisms of SLCO1B1 and ABCG2 to the pharmacokinetics of a dual substrate, pitavastatin, 2 mg of pitavastatin was administered to 38 healthy volunteers and pharmacokinetic parameters were compared among the following groups: 421C/C(*)1b/(*)1b (group 1), 421C/C(*)1b/(*)15 (group 2), 421C/C(*)15/(*)15 and 421C/A(*)15/(*)15 (group 3), 421C/A(*)1b/(*)1b (group 4), 421A/A(*)1b/(*)1b (group 5), and 421C/A(*)1b/(*)15 (group 6). In SLCO1B1, pitavastatin area under plasma concentration-time curve from 0 to 24 h (AUC(0-24)) for groups 1, 2, and 3 was 81.1+/-18.1, 144+/-32, and 250+/-57 ng h/ml, respectively, with significant differences among all three groups. In contrast to SLCO1B1, AUC(0-24) in groups 1, 4, and 5 was 81.1+/-18.1, 96.7+/-35.4, and 78.2+/-8.2 ng h/ml, respectively. Although the SLCO1B1 polymorphism was found to have a significant effect on the pharmacokinetics of pitavastatin, a nonsynonymous ABCG2 variant, 421C>A, did not appear to be associated with the altered pharmacokinetics of pitavastatin. PMID- 17460608 TI - Genetic variation in the renal sodium transporters NKCC2, NCC, and ENaC in relation to the effects of loop diuretic drugs. AB - There is little data on genetic predictors of loop diuretic efficacy in humans. Therefore, we investigated the diuretic effects of single oral doses of bumetanide, frusemide, and torsemide in a crossover study in 97 healthy Caucasians in relation to genetic variation in the renal sodium transporters NKCC2 (coded by SLC12A1), NCC (SLC12A3), and ENaC (three subunits coded by SCNN1A, SCNN1B, and SCNN1G). The NCC alanine 264 allele (Gly264Ala) and the most frequent SCNN1B haplotype were associated with stronger diuresis, indicating lower reabsorbing function of these alleles. The variant alleles of the tightly coupled polymorphisms rs5723 (Leu649Leu) and rs5729 in SCNN1G were associated with weaker diuresis, indicating higher activity. Extended haplotype homozygosity implied evolutionary selection of the NCC alanine 264 allele. In conclusion, acute diuretic effects of loop diuretics were affected by genetic variation in sodium transporters that, in the nephron, are located distally from NKCC2. PMID- 17460609 TI - Psychological stress can trigger atopic dermatitis in NC/Nga mice: an inhibitory effect of corticotropin-releasing factor. AB - Atopic dermatitis (AD) is one of the most common inflammatory diseases of the skin and is usually associated with a family history of atopic diathesis. It has been well established that many environmental or psychological factors aggravate AD. However, it is not clear whether psychological stress by itself can trigger AD. We examined the effect of psychological stress on the onset of AD, using an animal model, the NC/Nga mouse. The animals were exposed to the water avoidance stress (WAS) test to induce psychological stress. Additionally, we examined how corticotropin-releasing factor (CRF) affected the development of AD induced by psychological stress. Under specific pathogen-free (SPF) conditions, NC/Nga mice did not develop AD-like skin lesions. In contrast, NC/Nga mice exposed to psychological stress developed AD-like skin lesions along with elevated levels of serum immunoglobulin E even when kept under SPF conditions. The AD-like skin lesions induced by WAS were completely blocked by pretreating the animals with CRF. Our data indicate that a psychological factor is capable of eliciting AD like skin lesions in NC/Nga mice. It is possible that the inhibitory effect of CRF may be mediated by the functional modification of various cells that have CRF receptors. PMID- 17460610 TI - Alcohol preference influences the subthalamic nucleus control on motivation for alcohol in rats. AB - In addition to its role in motor and attentional processes, the subthalamic nucleus (STN) has also been recently demonstrated to be involved in motivational function. Indeed, bilateral STN lesions modulate differentially the motivation for natural rewards and drugs of abuse, increasing motivation for food and decreasing motivation for cocaine in rats. Here, we show that in outbred rats, the STN can modulate the motivation for alcohol according to alcohol preference, without affecting alcohol intake. When performed on 'High-Drinker' rats, STN lesions enhanced the breaking point (BP) under a progressive ratio schedule of reinforcement and increased the time spent in the environment previously paired with alcohol access in the place preference paradigm. In contrast, when performed on 'Low-Drinker' rats, STN lesions decreased the BP and increased the time spent in the environment paired with water. These results show that STN lesions enhance the motivation for alcohol in rats showing a high alcohol preference, whereas they decrease it in rats showing a low preference for alcohol. These results suggest that the STN plays a complex role in the reward circuit, that is not limited to a PMID- 17460611 TI - IMPA1 is essential for embryonic development and lithium-like pilocarpine sensitivity. AB - Lithium has been the standard pharmacological treatment for bipolar disorder over the last 50 years; however, the molecular targets through which lithium exerts its therapeutic effects are still not defined. We characterized the phenotype of mice with a dysfunctional IMPA1 gene (IMPA1-/-) to study the in vivo physiological functions of IMPA1, in general, and more specifically its potential role as a molecular target in mediating lithium-dependent physiological effects. Homozygote IMPA1-/- mice died in utero between days 9.5 and 10.5 post coitum (p.c.) demonstrating the importance of IMPA1 in early embryonic development. Intriguingly, the embryonic lethality could be reversed by myo-inositol supplementation via the pregnant mothers. In brains of adult IMPA1-/- mice, IMPase activity levels were found to be reduced (up to 65% in hippocampus); however, inositol levels were not found to be altered. Behavioral analysis of the IMPA1-/- mice indicated an increased motor activity in both the open-field test and the forced-swim test as well as a strongly increased sensitivity to pilocarpine-induced seizures, the latter supporting the idea that IMPA1 represents a physiologically relevant target for lithium. In conclusion the IMPA1 /- mouse represents a novel model to study inositol homeostasis, and indicates that genetic inactivation of IMPA1 can mimic some actions of lithium. PMID- 17460612 TI - Combined exposure to nicotine and ethanol in adolescent mice differentially affects anxiety levels during exposure, short-term, and long-term withdrawal. AB - Smoking and consumption of alcoholic beverages are frequently associated during adolescence. This association could be explained by the cumulative behavioral effects of nicotine and ethanol, particularly those related to anxiety levels. However, despite epidemiological findings, there have been few animal studies of the basic neurobiology of the combined exposure in the adolescent brain. In the present work we assessed, through the use of the elevated plus maze, the short- and long-term anxiety effects of nicotine (NIC) and/or ethanol (ETOH) exposure during adolescence (from the 30th to the 45th postnatal day) in four groups of male and female C57BL/6 mice: (1) Concomitant NIC (nicotine free-base solution (50 microg/ml) in 2% saccharin to drink) and ETOH (ethanol solution (25%, 2 g/kg) i.p. injected every other day) exposure; (2) NIC exposure; (3) ETOH exposure; (4) Vehicle. C57BL/6 mice were selected, in spite of the fact that they present slower ethanol metabolism, because they readily consume nicotine in the concentration used in the present study. During exposure (45th postnatal day: PN45), our results indicated that ethanol was anxiolytic in adolescent mice and that nicotine reverted this effect. Short-term drug withdrawal (PN50) elicited sex-dependent effects: exposure to nicotine and/or ethanol was anxiogenic only for females. Although neither nicotine nor ethanol effects persisted up to 1 month postexposure (PN75), the coadministration elicited an anxiogenic response. In spite of the fact that generalizations based on the results from a single strain of mice are prone to shortcomings, our results suggest that the deficient response to the anxiolytic effects of ethanol in adolescents co-exposed to nicotine may drive higher ethanol consumption. Additionally, increased anxiety during long-term smoking and drinking withdrawal may facilitate relapse to drug use. PMID- 17460613 TI - Ventral striatal blood flow is altered by acute nicotine but not withdrawal from nicotine. AB - Neural mechanisms underlying the reinforcing effects of nicotine and other drugs have been widely studied and are known to involve the ventral striatum, which is part of the mesocorticolimbic dopamine system. In contrast, mechanisms of nicotine withdrawal have received less attention although subjective withdrawal likely contributes to the difficulty of quitting. The goal of this study was to determine if nicotine withdrawal was associated with alterations of cerebral blood flow (CBF) in ventral striatum. Twelve smokers, moderately dependent on nicotine, underwent MR dynamic susceptibility contrast (DSC) imaging at baseline, after overnight withdrawal from nicotine, and after nicotine replacement. DSC images were used to calculate CBF in three regions of interest: ventral striatum, thalamus, and medial frontal cortex. Subjective withdrawal symptoms were measured at each time point. In spite of significant subjective withdrawal symptoms, there was no main effect of withdrawal on CBF in the three regions. However, there was a significant correlation between the increase in withdrawal symptoms and a reduction in thalamic CBF. In contrast to withdrawal, nicotine replacement significantly increased CBF in ventral striatum. Our findings are consistent with the known role of ventral striatum in drug reward. The lack of a main effect on withdrawal, but correlation of thalamic blood flow with withdrawal symptoms suggests that more complex mechanisms mediate the subjective features of the withdrawal state. PMID- 17460614 TI - Stimulation of the beta3-Adrenoceptor as a novel treatment strategy for anxiety and depressive disorders. AB - The characterization of the first selective orally active and brain-penetrant beta3-adrenoceptor agonist, SR58611A (amibegron), has opened new possibilities for exploring the involvement of this receptor in stress-related disorders. By using a battery of tests measuring a wide range of anxiety-related behaviors in rodents, including the mouse defense test battery, the elevated plus-maze, social interaction, stress-induced hyperthermia, four-plate, and punished drinking tests, we demonstrated for the first time that the stimulation of the beta3 receptor by SR58611A resulted in robust anxiolytic-like effects, with minimal active doses ranging from 0.3 to 10 mg/kg p.o., depending on the procedure. These effects paralleled those obtained with the prototypical benzodiazepine anxiolytic diazepam or chlordiazepoxide. Moreover, when SR58611A was tested in acute or chronic models of depression in rodents, such as the forced-swimming and the chronic mild stress tests, it produced antidepressant-like effects, which were comparable in terms of the magnitude of the effects to those of the antidepressant fluoxetine or imipramine. Supporting these behavioral data, SR58611A modified spontaneous sleep parameters in a manner comparable to that observed with fluoxetine. Importantly, SR58611A was devoid of side effects related to cognition (as shown in the Morris water maze and object recognition tasks), motor activity (in the rotarod), alcohol interaction, or physical dependence. Antagonism studies using pharmacological tools targeting a variety of neurotransmitters involved in anxiety and depression and the use of mice lacking the beta3 adrenoceptor suggested that these effects of SR58611A are mediated by beta3 adrenoceptors. Taken as a whole, these findings indicate that the pharmacological stimulation of beta3 adrenoceptors may represent an innovative approach for the treatment of anxiety and depressive disorders. PMID- 17460615 TI - Gene-by-environment (serotonin transporter and childhood maltreatment) interaction for anxiety sensitivity, an intermediate phenotype for anxiety disorders. AB - Anxiety sensitivity (AS) is a dispositional characteristic that predisposes to the development of anxiety disorders (eg, panic and post-traumatic stress disorder) and major depression. AS is subject to genetic and environmental influences, the former as yet unidentified and the latter known to include childhood maltreatment. The serotonin transporter gene (SLC6A4) promoter polymorphism (5-HTTLPR) has been associated with depression, but most consistently in the context of environmental stress. We tested the hypothesis that 5-HTTLPR genotype and childhood maltreatment would interact to increase susceptibility to AS in young adults. Subjects were European-American college undergraduates (N=150, median age 18 years) characterized on a measures of AS (Anxiety Sensitivity Index) and retrospective childhood maltreatment (Childhood Trauma Questionnaire [CTQ]). 5-HTTLPR genotypes were obtained from blood-derived DNA. Linear regression was used to model relationships between 5-HTTLPR, childhood emotional abuse, and AS; covariates such as sex, neuroticism, and ancestral proportion scores were incorporated into some models in a larger, ethnically heterogenous sample (N=247) to evaluate robustness of the findings to model assumptions. A statistically signficant interaction was observed between levels of childhood emotional (or physical) maltreatment and 5-HTTLPR genotype. Specifically, S/S individuals with higher levels of maltreatment had significantly higher levels of AS than subjects in other groups. No such relationship was found for neuroticism, attesting to the possible specificity of the findings for AS. Findings were consistently robust to the inclusion of covariates, and were not confounded by population stratification. In conclusion, these results provide evidence of a specific genetic influence on anxiety sensitivity-an intermediate phenotype for anxiety (and depressive) disorders; this effect is modified by severity of childhood maltreatment. These findings are consistent with the notion that 5-HTTLPR operates broadly to moderate emotional responsivity to stress. PMID- 17460616 TI - Haloperidol differentially modulates prepulse inhibition and p50 suppression in healthy humans stratified for low and high gating levels. AB - Schizophrenia patients exhibit deficits in sensory gating as indexed by reduced prepulse inhibition (PPI) and P50 suppression, which have been linked to psychotic symptom formation and cognitive deficits. Although recent evidence suggests that atypical antipsychotics might be superior over typical antipsychotics in reversing PPI and P50 suppression deficits not only in schizophrenia patients, but also in healthy volunteers exhibiting low levels of PPI, the impact of typical antipsychotics on these gating measures is less clear. To explore the impact of the dopamine D2-like receptor system on gating and cognition, the acute effects of haloperidol on PPI, P50 suppression, and cognition were assessed in 26 healthy male volunteers split into subgroups having low vs high PPI or P50 suppression levels using a placebo-controlled within subject design. Haloperidol failed to increase PPI in subjects exhibiting low levels of PPI, but attenuated PPI in those subjects with high sensorimotor gating levels. Furthermore, haloperidol increased P50 suppression in subjects exhibiting low P50 gating and disrupted P50 suppression in individuals expressing high P50 gating levels. Independently of drug condition, high PPI levels were associated with superior strategy formation and execution times in a subset of cognitive tests. Moreover, haloperidol impaired spatial working memory performance and planning ability. These findings suggest that dopamine D2-like receptors are critically involved in the modulation of P50 suppression in healthy volunteers, and to a lesser extent also in PPI among subjects expressing high sensorimotor gating levels. Furthermore, the results suggest a relation between sensorimotor gating and working memory performance. PMID- 17460621 TI - Taking the first step. PMID- 17460622 TI - The university of the future. PMID- 17460617 TI - Assessment of cognitive brain function in ecstasy users and contributions of other drugs of abuse: results from an FMRI study. AB - Heavy ecstasy use has been associated with neurocognitive deficits in various behavioral and brain imaging studies. However, this association is not conclusive owing to the unavoidable confounding factor of polysubstance use. The present study, as part of the Netherlands XTC Toxicity study, investigated specific effects of ecstasy on working memory, attention, and associative memory, using functional magnetic resonance imaging (fMRI). A large sample (n=71) was carefully composed based on variation in the amount and type of drugs that were used. The sample included 33 heavy ecstasy users (mean 322 pills lifetime). Neurocognitive brain function in three domains: working memory, attention, and associative memory, was assessed with performance measures and fMRI. Independent effects of the use of ecstasy, amphetamine, cocaine, cannabis, alcohol, tobacco, and of gender and IQ were assessed and separated by means of multiple regression analyses. Use of ecstasy had no effect on working memory and attention, but drug use was associated with reduced associative memory performance. Multiple regression analysis showed that associative memory performance was affected by amphetamine much more than by ecstasy. Both drugs affected associative memory related brain activity, but the effects were consistently in opposite directions, suggesting that different mechanisms are at play. This could be related to the different neurotransmitter systems these drugs predominantly act upon, that is, serotonin (ecstasy) vs dopamine (amphetamine) systems. PMID- 17460623 TI - Conflicted contractors. PMID- 17460625 TI - China struggles to square growth and emissions. PMID- 17460626 TI - Primate work faces German veto. PMID- 17460627 TI - Brain's speech site is revisited and revised. PMID- 17460628 TI - Elusive flowering signal pruned of mystery at last. PMID- 17460630 TI - Online resources threaten livelihood of libraries. PMID- 17460631 TI - Regulators pull contract for chemical review. PMID- 17460635 TI - Nanotech growing pains. PMID- 17460632 TI - Rival genetics projects build bridges. PMID- 17460636 TI - Immunology: Pimp my antibody. PMID- 17460637 TI - US higher education: the Arizona experiment. PMID- 17460638 TI - US higher education: medicinal properties. PMID- 17460639 TI - Millennium: big effort has produced statistical results. PMID- 17460640 TI - Millennium: invest in country statistical systems. PMID- 17460641 TI - A logged forest in Borneo is better than none at all. PMID- 17460642 TI - When good drugs go bad. PMID- 17460647 TI - Disappearing act. PMID- 17460648 TI - Plant ecology: resourceful invaders. PMID- 17460649 TI - Astrophysics: the answer is blowing in the wind. PMID- 17460650 TI - Biogeochemistry: iron findings. PMID- 17460652 TI - Atmospheric chemistry: a forest air of chirality. PMID- 17460653 TI - Computing: the security of knowing nothing. PMID- 17460654 TI - Chemical biology: dressed-up proteins. PMID- 17460655 TI - Applied physics: weight inside. PMID- 17460656 TI - Mathematical physics: added dimensions to grain growth. PMID- 17460657 TI - Stem cells: blood lines from embryo to adult. PMID- 17460658 TI - Obituary: John Backus (1924-2007). PMID- 17460660 TI - Chemical glycosylation in the synthesis of glycoconjugate antitumour vaccines. AB - Therapeutic vaccines derived from carbohydrate antigen-adjuvant combinations are a promising approach for cancer immunotherapy. One of the critical limitations in this area is access to sufficient quantities of tumour-associated carbohydrate antigens and glycoconjugate adjuvants. At present, availability of the complex oligosaccharide constructs that are needed for the systematic design and evaluation of novel vaccine formulations relies on de novo chemical synthesis. The use of both state-of-the-art and emerging glycosylation technologies has led to significant advances in this field, allowing the clinical exploration of carbohydrate-based antigens in the treatment of cancer. PMID- 17460661 TI - Unusual sugar biosynthesis and natural product glycodiversification. AB - The enzymes involved in the biosynthesis of carbohydrates and the attachment of sugar units to biological acceptor molecules catalyse an array of chemical transformations and coupling reactions. In prokaryotes, both common sugar precursors and their enzymatically modified derivatives often become substituents of biologically active natural products through the action of glycosyltransferases. Recently, researchers have begun to harness the power of these biological catalysts to alter the sugar structures and glycosylation patterns of natural products both in vivo and in vitro. Biochemical and structural studies of sugar biosynthetic enzymes and glycosyltransferases, coupled with advances in bioengineering methodology, have ushered in a new era of drug development. PMID- 17460662 TI - Cycling of O-linked beta-N-acetylglucosamine on nucleocytoplasmic proteins. AB - All animals and plants dynamically attach and remove O-linked beta-N acetylglucosamine (O-GlcNAc) at serine and threonine residues on myriad nuclear and cytoplasmic proteins. O-GlcNAc cycling, which is tightly regulated by the concerted actions of two highly conserved enzymes, serves as a nutrient and stress sensor. On some proteins, O-GlcNAc competes directly with phosphate for serine/threonine residues. Glycosylation with O-GlcNAc modulates signalling, and influences protein expression, degradation and trafficking. Emerging data indicate that O-GlcNAc glycosylation has a role in the aetiology of diabetes and neurodegeneration. PMID- 17460663 TI - Glycan-based interactions involving vertebrate sialic-acid-recognizing proteins. AB - All cells in nature are covered by a dense and complex array of carbohydrates. Given their prominence on cell surfaces, it is not surprising that these glycans mediate and/or modulate many cellular interactions. Proteins that bind sialic acid, a sugar that is found on the surface of the cell and on secreted proteins in vertebrates, are involved in a broad range of biological processes, including intercellular adhesion, signalling and microbial attachment. Studying the roles of such proteins in vertebrates has improved our understanding of normal physiology, disease and human evolution. PMID- 17460664 TI - Heparan sulphate proteoglycans fine-tune mammalian physiology. AB - Heparan sulphate proteoglycans reside on the plasma membrane of all animal cells studied so far and are a major component of extracellular matrices. Studies of model organisms and human diseases have demonstrated their importance in development and normal physiology. A recurrent theme is the electrostatic interaction of the heparan sulphate chains with protein ligands, which affects metabolism, transport, information transfer, support and regulation in all organ systems. The importance of these interactions is exemplified by phenotypic studies of mice and humans bearing mutations in the core proteins or the biosynthetic enzymes responsible for assembling the heparan sulphate chains. PMID- 17460665 TI - Exploiting the defensive sugars of HIV-1 for drug and vaccine design. AB - The sustained effort towards developing an antibody vaccine against HIV/AIDS has provided much of our understanding of viral immunology. It is generally accepted that one of the main barriers to antibody neutralization of HIV is the array of protective structural carbohydrates that covers the antigens on the virus's surface. Intriguingly, however, recent findings suggest that these carbohydrates, which have evolved to protect HIV and promote its transmission, are also attractive therapeutic targets. PMID- 17460666 TI - Synthesis and medical applications of oligosaccharides. AB - Our understanding of the different glycoconjugates present on cells, proteins and entire organisms is lagging far behind advances in genomics and proteomics. Carbohydrate sequencing and the synthesis of defined oligosaccharides are two key technologies that have contributed to progress in glycomics research. Synthetic tools and high-throughput experiments such as carbohydrate arrays are beginning to affect biological research. These techniques are now being applied to the development of carbohydrate-based diagnostics, vaccines and therapeutics. PMID- 17460667 TI - The von Neumann relation generalized to coarsening of three-dimensional microstructures. AB - Cellular structures or tessellations are ubiquitous in nature. Metals and ceramics commonly consist of space-filling arrays of single-crystal grains separated by a network of grain boundaries, and foams (froths) are networks of gas-filled bubbles separated by liquid walls. Cellular structures also occur in biological tissue, and in magnetic, ferroelectric and complex fluid contexts. In many situations, the cell/grain/bubble walls move under the influence of their surface tension (capillarity), with a velocity proportional to their mean curvature. As a result, the cells evolve and the structure coarsens. Over 50 years ago, von Neumann derived an exact formula for the growth rate of a cell in a two-dimensional cellular structure (using the relation between wall velocity and mean curvature, the fact that three domain walls meet at 120 degrees and basic topology). This forms the basis of modern grain growth theory. Here we present an exact and much-sought extension of this result into three (and higher) dimensions. The present results may lead to the development of predictive models for capillarity-driven microstructure evolution in a wide range of industrial and commercial processing scenarios--such as the heat treatment of metals, or even controlling the 'head' on a pint of beer. PMID- 17460668 TI - Chlorine isotope homogeneity of the mantle, crust and carbonaceous chondrites. AB - Chlorine in the Earth is highly depleted relative to carbonaceous chondrites and solar abundances. Knowledge of the Cl concentrations and distribution on Earth is essential for understanding the origin of these depletions. Large differences in the stable chlorine isotope ratios of meteoritic, mantle and crustal materials have been used as evidence for distinct reservoirs in the solar nebula and to calculate the relative proportions of Cl in the mantle and crust. Here we report that large isotopic differences do not exist, and that carbonaceous chondrites, mantle and crust all have the same 37Cl/35Cl ratios. We have further analysed crustal sediments from the early Archaean era to the Recent epoch and find no systematic isotopic variations with age, demonstrating that the mantle and crust have always had the same delta37Cl value. The similarity of mantle, crust and carbonaceous chondrites establishes that there were no nebular reservoirs with distinct isotopic compositions, no isotopic fractionation during differentiation of the Earth and no late (post-core formation) Cl-bearing volatile additions to the crustal veneer with a unique isotopic composition. PMID- 17460669 TI - Weighing of biomolecules, single cells and single nanoparticles in fluid. AB - Nanomechanical resonators enable the measurement of mass with extraordinary sensitivity. Previously, samples as light as 7 zeptograms (1 zg = 10(-21) g) have been weighed in vacuum, and proton-level resolution seems to be within reach. Resolving small mass changes requires the resonator to be light and to ring at a very pure tone-that is, with a high quality factor. In solution, viscosity severely degrades both of these characteristics, thus preventing many applications in nanotechnology and the life sciences where fluid is required. Although the resonant structure can be designed to minimize viscous loss, resolution is still substantially degraded when compared to measurements made in air or vacuum. An entirely different approach eliminates viscous damping by placing the solution inside a hollow resonator that is surrounded by vacuum. Here we demonstrate that suspended microchannel resonators can weigh single nanoparticles, single bacterial cells and sub-monolayers of adsorbed proteins in water with sub-femtogram resolution (1 Hz bandwidth). Central to these results is our observation that viscous loss due to the fluid is negligible compared to the intrinsic damping of our silicon crystal resonator. The combination of the low resonator mass (100 ng) and high quality factor (15,000) enables an improvement in mass resolution of six orders of magnitude over a high-end commercial quartz crystal microbalance. This gives access to intriguing applications, such as mass based flow cytometry, the direct detection of pathogens, or the non-optical sizing and mass density measurement of colloidal particles. PMID- 17460670 TI - Effect of natural iron fertilization on carbon sequestration in the Southern Ocean. AB - The availability of iron limits primary productivity and the associated uptake of carbon over large areas of the ocean. Iron thus plays an important role in the carbon cycle, and changes in its supply to the surface ocean may have had a significant effect on atmospheric carbon dioxide concentrations over glacial interglacial cycles. To date, the role of iron in carbon cycling has largely been assessed using short-term iron-addition experiments. It is difficult, however, to reliably assess the magnitude of carbon export to the ocean interior using such methods, and the short observational periods preclude extrapolation of the results to longer timescales. Here we report observations of a phytoplankton bloom induced by natural iron fertilization--an approach that offers the opportunity to overcome some of the limitations of short-term experiments. We found that a large phytoplankton bloom over the Kerguelen plateau in the Southern Ocean was sustained by the supply of iron and major nutrients to surface waters from iron-rich deep water below. The efficiency of fertilization, defined as the ratio of the carbon export to the amount of iron supplied, was at least ten times higher than previous estimates from short-term blooms induced by iron-addition experiments. This result sheds new light on the effect of long-term fertilization by iron and macronutrients on carbon sequestration, suggesting that changes in iron supply from below--as invoked in some palaeoclimatic and future climate change scenarios--may have a more significant effect on atmospheric carbon dioxide concentrations than previously thought. PMID- 17460671 TI - The role of fluids in lower-crustal earthquakes near continental rifts. AB - The occurrence of earthquakes in the lower crust near continental rifts has long been puzzling, as the lower crust is generally thought to be too hot for brittle failure to occur. Such anomalous events have usually been explained in terms of the lower crust being cooler than normal. But if the lower crust is indeed cold enough to produce earthquakes, then the uppermost mantle beneath it should also be cold enough, and yet uppermost mantle earthquakes are not observed. Numerous lower-crustal earthquakes occur near the southwestern termination of the Taupo Volcanic Zone (TVZ), an active continental rift in New Zealand. Here we present three-dimensional tomographic imaging of seismic velocities and seismic attenuation in this region using data from a dense seismograph deployment. We find that crustal earthquakes accurately relocated with our three-dimensional seismic velocity model form a continuous band along the rift, deepening from mostly less than 10 km in the central TVZ to depths of 30-40 km in the lower crust, 30 km southwest of the termination of the volcanic zone. These earthquakes often occur in swarms, suggesting fluid movement in critically loaded fault zones. Seismic velocities within the band are also consistent with the presence of fluids, and the deepening seismicity parallels the boundary between high seismic attenuation (interpreted as partial melt) within the central TVZ and low seismic attenuation in the crust to the southwest. This linking of upper and lower-crustal seismicity and crustal structure allows us to propose a common explanation for all the seismicity, involving the weakening of faults on the periphery of an otherwise dry, mafic crust by hot fluids, including those exsolved from underlying melt. Such fluids may generally be an important driver of lower-crustal seismicity near continental rifts. PMID- 17460672 TI - Resource-use efficiency and plant invasion in low-resource systems. AB - No species can maximize growth, reproduction and competitive ability across all environments, so the success of invasive species is habitat-dependent. Nutrient rich habitats often experience more invasion than resource-poor habitats, a pattern consistent with traits generally associated with successful invaders (high growth rates, early reproduction and many offspring). However, invaders do colonize resource-poor environments, and the mechanisms that allow their success in these systems are poorly understood. Traits associated with resource conservation are widespread among species adapted to resource-poor environments, and invasive species may succeed in low-resource environments by employing resource conservation traits such as high resource-use efficiency (RUE; carbon assimilation per unit of resource). We investigated RUE in invasive and native species from three habitats in Hawaii where light, water or nutrient availability was limiting to plant growth. Here we show that across multiple growth forms and broad taxonomic diversity invasive species were generally more efficient than native species at using limiting resources on short timescales and were similarly efficient when RUE measures were integrated over leaf lifespans. Our data challenge the idea that native species generally outperform invasive species under conditions of low resource availability, and suggest that managing resource levels is not always an effective strategy for invasive species control. PMID- 17460673 TI - How swifts control their glide performance with morphing wings. AB - Gliding birds continually change the shape and size of their wings, presumably to exploit the profound effect of wing morphology on aerodynamic performance. That birds should adjust wing sweep to suit glide speed has been predicted qualitatively by analytical glide models, which extrapolated the wing's performance envelope from aerodynamic theory. Here we describe the aerodynamic and structural performance of actual swift wings, as measured in a wind tunnel, and on this basis build a semi-empirical glide model. By measuring inside and outside swifts' behavioural envelope, we show that choosing the most suitable sweep can halve sink speed or triple turning rate. Extended wings are superior for slow glides and turns; swept wings are superior for fast glides and turns. This superiority is due to better aerodynamic performance-with the exception of fast turns. Swept wings are less effective at generating lift while turning at high speeds, but can bear the extreme loads. Finally, our glide model predicts that cost-effective gliding occurs at speeds of 8-10 m s(-1), whereas agility related figures of merit peak at 15-25 m s(-1). In fact, swifts spend the night ('roost') in flight at 8-10 m s(-1) (ref. 11), thus our model can explain this choice for a resting behaviour. Morphing not only adjusts birds' wing performance to the task at hand, but could also control the flight of future aircraft. PMID- 17460674 TI - Opioids block long-term potentiation of inhibitory synapses. AB - Excitatory brain synapses are strengthened or weakened in response to specific patterns of synaptic activation, and these changes in synaptic strength are thought to underlie persistent pathologies such as drug addiction, as well as learning. In contrast, there are few examples of synaptic plasticity of inhibitory GABA (gamma-aminobutyric acid)-releasing synapses. Here we report long term potentiation of GABA(A)-mediated synaptic transmission (LTP(GABA)) onto dopamine neurons of the rat brain ventral tegmental area, a region required for the development of drug addiction. This novel form of LTP is heterosynaptic, requiring postsynaptic NMDA (N-methyl-d-aspartate) receptor activation at glutamate synapses, but resulting from increased GABA release at neighbouring inhibitory nerve terminals. NMDA receptor activation produces nitric oxide, a retrograde signal released from the postsynaptic dopamine neuron. Nitric oxide initiates LTP(GABA) by activating guanylate cyclase in GABA-releasing nerve terminals. Exposure to morphine both in vitro and in vivo prevents LTP(GABA). Whereas brief treatment with morphine in vitro blocks LTP(GABA) by inhibiting presynaptic glutamate release, in vivo exposure to morphine persistently interrupts signalling from nitric oxide to guanylate cyclase. These neuroadaptations to opioid drugs might contribute to early stages of addiction, and may potentially be exploited therapeutically using drugs targeting GABA(A) receptors. PMID- 17460675 TI - Expanding the diversity of chemical protein modification allows post translational mimicry. AB - One of the most important current scientific paradoxes is the economy with which nature uses genes. In all higher animals studied, we have found many fewer genes than we would have previously expected. The functional outputs of the eventual products of genes seem to be far more complex than the more restricted blueprint. In higher organisms, the functions of many proteins are modulated by post translational modifications (PTMs). These alterations of amino-acid side chains lead to higher structural and functional protein diversity and are, therefore, a leading contender for an explanation for this seeming incongruity. Natural protein production methods typically produce PTM mixtures within which function is difficult to dissect or control. Until now it has not been possible to access pure mimics of complex PTMs. Here we report a chemical tagging approach that enables the attachment of multiple modifications to bacterially expressed (bare) protein scaffolds: this approach allows reconstitution of functionally effective mimics of higher organism PTMs. By attaching appropriate modifications at suitable distances in the widely-used LacZ reporter enzyme scaffold, we created protein probes that included sensitive systems for detection of mammalian brain inflammation and disease. Through target synthesis of the desired modification, chemistry provides a structural precision and an ability to retool with a chosen PTM in a manner not available to other approaches. In this way, combining chemical control of PTM with readily available protein scaffolds provides a systematic platform for creating probes of protein-PTM interactions. We therefore anticipate that this ability to build model systems will allow some of this gene product complexity to be dissected, with the aim of eventually being able to completely duplicate the patterns of a particular protein's PTMs from an in vivo assay into an in vitro system. PMID- 17460676 TI - MicroRNAs as tumor suppressors. PMID- 17460678 TI - Reactive oxygen species and the segregation of mtDNA sequence variants. PMID- 17460680 TI - Heritable germline epimutations in humans. PMID- 17460681 TI - Inherited epimutation or a haplotypic basis for the propensity to silence? PMID- 17460682 TI - Heritable germline epimutation is not the same as transgenerational epigenetic inheritance. PMID- 17460686 TI - Multiple prostate cancer risk variants on 8q24. PMID- 17460687 TI - Road rage and fruit flies. PMID- 17460688 TI - A quantitative trait locus regulating rice grain width. PMID- 17460691 TI - Drosophila researchers focus on human disease. PMID- 17460694 TI - FANCI is a second monoubiquitinated member of the Fanconi anemia pathway. AB - Activation of the Fanconi anemia (FA) DNA damage-response pathway results in the monoubiquitination of FANCD2, which is regulated by the nuclear FA core ubiquitin ligase complex. A FANCD2 protein sequence-based homology search facilitated the discovery of FANCI, a second monoubiquitinated component of the FA pathway. Biallelic mutations in the gene coding for this protein were found in cells from four FA patients, including an FA-I reference cell line. PMID- 17460695 TI - Structural basis of Na(+)/K(+)-ATPase adaptation to marine environments. AB - Throughout evolution, enzymes have adapted to perform in different environments. The Na(+)/K(+) pump, an enzyme crucial for maintaining ionic gradients across cell membranes, is strongly influenced by the ionic environment. In vertebrates, the pump sees much less external Na(+) (100-160 mM) than it does in osmoconformers such as squid (450 mM), which live in seawater. If the extracellular architecture of the squid pump were identical to that of vertebrates, then at the resting potential, the pump's function would be severely compromised because the negative voltage would drive Na(+) ions back to their binding sites, practically abolishing forward transport. Here we show that four amino acids that ring the external mouth of the ion translocation pathway are more positive in squid, thereby reducing the pump's sensitivity to external Na(+) and explaining how it can perform optimally in the marine environment. PMID- 17460696 TI - Essential function of the built-in lid in the allosteric regulation of eukaryotic and archaeal chaperonins. AB - Chaperonins are allosteric double-ring ATPases that mediate cellular protein folding. ATP binding and hydrolysis control opening and closing of the central chaperonin chamber, which transiently provides a protected environment for protein folding. During evolution, two strategies to close the chaperonin chamber have emerged. Archaeal and eukaryotic group II chaperonins contain a built-in lid, whereas bacterial chaperonins use a ring-shaped cofactor as a detachable lid. Here we show that the built-in lid is an allosteric regulator of group II chaperonins, which helps synchronize the subunits within one ring and, to our surprise, also influences inter-ring communication. The lid is dispensable for substrate binding and ATP hydrolysis, but is required for productive substrate folding. These regulatory functions of the lid may serve to allow the symmetrical chaperonins to function as 'two-stroke' motors and may also provide a timer for substrate encapsulation within the closed chamber. PMID- 17460697 TI - A variant in CDKAL1 influences insulin response and risk of type 2 diabetes. AB - We conducted a genome-wide association study for type 2 diabetes (T2D) in Icelandic cases and controls, and we found that a previously described variant in the transcription factor 7-like 2 gene (TCF7L2) gene conferred the most significant risk. In addition to confirming two recently identified risk variants, we identified a variant in the CDKAL1 gene that was associated with T2D in individuals of European ancestry (allele-specific odds ratio (OR) = 1.20 (95% confidence interval, 1.13-1.27), P = 7.7 x 10(-9)) and individuals from Hong Kong of Han Chinese ancestry (OR = 1.25 (1.11-1.40), P = 0.00018). The genotype OR of this variant suggested that the effect was substantially stronger in homozygous carriers than in heterozygous carriers. The ORs for homozygotes were 1.50 (1.31 1.72) and 1.55 (1.23-1.95) in the European and Hong Kong groups, respectively. The insulin response for homozygotes was approximately 20% lower than for heterozygotes or noncarriers, suggesting that this variant confers risk of T2D through reduced insulin secretion. PMID- 17460698 TI - Prevalence of major depressive disorder in self-referred patients in a late onset hypogonadism clinic. AB - The goal was to clarify the rate of major depressive disorder (MDD) in self referred patients who visited our late onset hypogonadism (LOH) clinic and the importance of screening of MDD in LOH clinic. Two hundred consecutive self referred patients who visited our LOH clinic were evaluated. MDD was diagnosed using, the Mini International Neuropsychiatric Interview (MINI) questionnaire. Scores of the Aging Males Symptom (AMS) scale were compared between the MDD and non-MDD groups with and without low testosterone values. Forty-two percent of all patients were categorized into the MDD group. Only 4% of the patients were considered to be LOH. The MDD group had significantly higher scores on the AMS scale than the LOH and non-MDD with normal testosterone groups. In conclusion, significant numbers of undiagnosed MDD patients visited the LOH clinic. Screening for MDD is an essential step in the LOH outpatient clinic. PMID- 17460699 TI - Potential differentiation of human mesenchymal stem cell transplanted in rat corpus cavernosum toward endothelial or smooth muscle cells. AB - One of the causes of erectile dysfunction (ED) is the damaged penile cavernous smooth muscle cells (SMCs) and sinus endothelial cells (ECs). To investigate the feasibility of applying immortalized human mesenchymal stem cells (MSCs) to penile cavernous ECs or SMCs repair in the treatment of ED, the in vivo potential differentiation of the immortalized human MSCs toward penile cavernous endothelial or smooth muscle was investigated. One clone of immortalized human bone marrow mesenchymal stem cell line B10 cells via retroviral vector encoding v myc were transplanted into the cavernosum of the Sprague-Dawley rats and harvested 2 weeks later. The expression of CD31, von Willebrand factor (vWF), smooth muscle cell actin (SMA), calponin and desmin was determined immunohistochemically in rat penile cavernosum. Multipotency of B10 to adipogenic, osteogenic or chondrogenic differentiation was found. Expression of EC specific markers (CD31 or vWF protein) and expression of SMC specific markers (calponin, SMA or desmin protein) were demonstrated in grafted B10 cells. When human MSCs were transplanted into the penile cavernosum, they have the potential to differentiate toward ECs or SMCs. Human MSCs may be a good candidate in the treatment of penile cavernosum injury. PMID- 17460700 TI - ABT-737, an inhibitor of Bcl-2 family proteins, is a potent inducer of apoptosis in multiple myeloma cells. AB - Disruption of pathways leading to programmed cell death plays a major role in most malignancies, including multiple myeloma (MM). ABT-737 is a BH3 mimetic small-molecule inhibitor that binds with high affinity to Bcl-2 and Bcl-xL, preventing the sequestration of proapoptotic molecules and shifting the cell survival/apoptosis balance toward apoptosis induction. In this study, we show that ABT-737 is cytotoxic to MM cell lines, including those resistant to conventional therapies, and primary tumor cells. Flow cytometric analysis of intracellular levels of Bcl-2 family proteins demonstrates a clear inversion of the Bax/Bcl-2 ratio leading to induction of apoptosis. Activation of the mitochondrial apoptosis pathway was indicated by mitochondrial membrane depolarization and caspase cleavage. Additionally, several signaling pathways known to be important for MM cell survival are disrupted following treatment with ABT-737. The impact of ABT-737 on survival could not be overcome by the addition of interleukin-6, vascular endothelial growth factor or insulin-like growth factor, suggesting that ABT-737 may be effective in preventing the growth and survival signals provided by the microenvironment. These data indicate that therapies targeting apoptotic pathways may be effective in MM treatment and warrant clinical evaluation of ABT-737 and similar drugs alone or in combination with other agents in the setting of MM. PMID- 17460701 TI - Childhood secondary ALL after ALL treatment. AB - Data on secondary acute lymphoblastic leukaemia (sALL) following ALL treatment are very rare. However, the incidence might be underestimated as sALLs without a significant lineage shift might automatically be diagnosed as relapses. Examination of immunoglobulin and T-cell receptor gene rearrangements brought a new tool that can help in discrimination between relapse and sALL. We focused on the recurrences of childhood ALL to discover the real frequency of the sALL after ALL treatment. We compared clonal markers in matched presentation and recurrence samples of 366 patients treated according to the Berlin-Frankfurt-Munster (BFM) based protocols. We found two cases of sALL and another three, where the recurrence is suspicious of being sALL rather than relapse. Our proposal for the 'secondary ALL after ALL' diagnostic criteria is as follows: (A) No clonal relationship between diagnosis and recurrence; (B) significant immunophenotypic shift--significant cytogenetic shift--gain/loss of a fusion gene. For the sALL (A) plus at least one (B) criterion should be fulfilled. With these criteria, the estimated frequency of the sALL after ALL is according to our data 0.5-1.5% of ALL recurrences on BFM-based protocols. Finally, we propose a treatment strategy for the patients with secondary disease. PMID- 17460702 TI - Effect of spacing intravenous bisphosphonates in patients with multiple myeloma in plateau phase. PMID- 17460703 TI - Treatment of severe autoimmune hemolytic anemia in B-cell chronic lymphocytic leukemia with alemtuzumab. PMID- 17460704 TI - Novel multi-parameter flow cytometry sensitively detects phenotypically distinct plasma cell subsets in plasma cell proliferative disorders. PMID- 17460705 TI - Lenalidomide therapy in del(5)(q31)-associated myelofibrosis: cytogenetic and JAK2V617F molecular remissions. PMID- 17460706 TI - JAK2V617F prevalence and allele burden in non-splanchnic venous thrombosis in the absence of overt myeloproliferative disorder. PMID- 17460707 TI - Monosomy 7 in myeloid malignancies: parental origin and monitoring by real-time quantitative PCR. PMID- 17460708 TI - Detecting the JAK2 V617F mutation in fresh and 'historic' blood and bone marrow. PMID- 17460709 TI - Blood pressure, antihypertensive treatment and factors associated with good blood pressure control in hypertensive diabetics: the Tarmidas study. AB - Numerous population studies confirm the high prevalence of hypertension in type II diabetic (DM2) subjects and that intensive antihypertensive treatment is more beneficial to diabetic than to nondiabetic hypertensive subjects, yet not many of these are specific to Spain. To assess the degree of blood pressure (BP) control and the effects of antihypertensive drugs in the medical management of hypertension in diabetic patients in specialist care centres throughout Spain, we studied the socio-demographic, clinical and relevant laboratory parameters of 796 hypertensive patients with DM2 (mean age 66.09 (95% confidence interval (CI): 64.08-68.10). The percentage of diabetic patients responding positively to BP control measures was lower when compared to the nondiabetic population in both Spain and Europe. The degree of control was poorer for systolic than for diastolic BP, yet 40.6% of the patients were only on monotherapy. The fact that antihypertensive treatment was modified in only 40% of the poorly controlled patients was also highly significant and could be attributed to a nonstringent use of clinical guidelines. Among the other differences between well-controlled and poorly controlled patients, we found that well-controlled patients presented with lower levels of cholesterol and triglycerides, a lower prevalence of excess weight/obesity, and a greater prevalence of cardiovascular and/or cerebrovascular disease despite having a greater percentage of patients on antiplatelet therapy. Better application of therapeutic guidelines and the prevention and treatment of compounding factors could improve the response rate to BP control measures in poorly controlled patients. PMID- 17460710 TI - Acute effects of renin-angiotensin system blockade on arterial function in hypertensive patients. AB - The acute effects of the renin-angiotensin system (RAS) blockers may be important in some clinical settings. To assess the acute impact of such drugs on arterial function, we studied the effects of captopril 25 mg, quinapril 20 mg and telmisartan 80 mg on 100 hypertensive patients, according to a randomized, double blind, placebo-controlled study. Central (aortic) blood pressure (BP) and augmentation index (AIx, a measure of wave reflections), as well as flow-mediated dilatation (FMD) of the brachial artery and forearm blood flow (FBF) (measures of conduit and resistance artery endothelial function, respectively), were evaluated before and 2 h after oral drug administration. Compared to placebo, captopril and quinapril decreased central systolic (by 7.5 mm Hg, P<0.05 and by 12.3 mm Hg, P<0.001) and diastolic BP (by 4.9 mm Hg, P<0.01 and by 8.4 mm Hg, P<0.001), whereas telmisartan had no significant effect (P=NS). Additionally, AIx was reduced after quinapril (absolute decrease of 7.2%, P<0.01) and marginally after captopril (decrease of 4.7%, P=0.07). Only quinapril led to a beneficial change of FMD (absolute increase of 2.7%, P<0.001). No treatment was related to significant changes of peak hyperaemic or 3-min hyperaemic FBF. In adjusted analyses, all the favourable alterations induced by quinapril were independent of potential confounding haemodynamic factors. Our data show that acute RAS inhibition with quinapril (20 mg) may be more beneficial in terms of arterial function and central haemodynamics compared to captopril (25 mg) or telmisartan (80 mg). Further studies are needed to investigate whether these acute arterial effects of quinapril are clinically significant. PMID- 17460711 TI - Plasma N terminal pro-brain natriuretic peptide levels and its determinants in a multi-ethnic population. AB - This study documents the determinants and plasma levels of N-terminal pro-brain natriuretic peptide (NT-proBNP) among hypertensive and normotensive subjects in a multi-ethnic population in the United Arab Emirates (UAE). We obtained demographic, anthropometric and clinical data, together with fasting NT-proBNP and biochemical indices from 128 hypertensive patients and 138 normotensive subjects matched for age, gender and ethnicity. Plasma NT-proBNP levels were significantly (P<0.001), and several-fold higher among hypertensives (median 5.92, inter quartile range (IQR): 1.79-18.48 pmol/l) than normotensives (median 1.78, IQR: 0.59-4.32 pmol/l) in the total study population, and the same was true for the ethnic groups separately. Similarly, plasma levels of glucose, blood urea nitrogen (BUN) and creatinine, but not insulin, were significantly (P<0.05) higher among hypertensives than normotensives. For all subjects combined, log NT proBNP correlated positively and significantly with age (P<0.01), log glucose (P<0.05), systolic blood pressure (SBP, P<0.001), log BUN (P<0.001) and log creatinine (P<0.001). Multivariate regression analysis showed that NT-proBNP levels were independently and positively correlated with SBP, age, gender, log BUN, Emirati and South East Asian ethnic groups and inversely associated with current exercise. In conclusion, we found circulating levels of NT-proBNP to be significantly increased in hypertensive versus normotensive subjects in the UAE and independently related to SBP, age, gender, indices of renal function and possibly exercise. Our results further suggest a possible modulating effect of ethnicity on NT-proBNP levels. PMID- 17460712 TI - The interarm blood pressure difference as predictor of cardiovascular events in patients with hypertension in primary care: cohort study. AB - Objectives of this study were to measure the prevalence of a difference in blood pressure (BP) between arms and determine whether a difference is associated with increased risk of cardiovascular events or death. A prospective cohort study of 247 patients with hypertension was undertaken in one rural general practice in England. The main outcome measures were mean difference in BP between arms and new episodes of myocardial infarction, cerebrovascular event, onset of angina or peripheral vascular disease or death. A total of 57/247 (23%) patients had a mean difference in systolic BP between arms of >or=10 mm Hg and 8/247 (3%) had a mean difference of >or=20 mm Hg. A total of 15/247 (6%) patients had a mean difference in diastolic BP between arms of >or=10 mm Hg. Survival analysis after 4.7 years (range 3.3-5.9) showed a shorter mean survival time without event or death for patients with a difference in systolic BP of >or=10 mm Hg compared with a difference of <10 mm Hg (3.7 (95% confidence interval, 3.2-4.2) versus 4.8 (4.6 5.1) years; P<0.001; hazard ratio 2.5 (1.5-4.2), P=0.001). Difference in systolic BP of >or=10 mm Hg between arms is common in this primary care population and is associated with a shorter survival time to death or new cardiovascular event. Detection of a difference between arms may identify hypertensive patients at increased risk of cardiovascular events. Such an approach would allow more effective targeting of resources in primary prevention strategies. PMID- 17460713 TI - Effects of lifestyle changes and metformin on salt sensitivity and nitric oxide metabolism in obese salt-sensitive Hispanics. AB - Salt sensitivity is associated with obesity, and increased cardiovascular morbidity and mortality. We investigated whether treatment of obesity and its associated metabolic abnormalities corrects salt sensitivity and restores impaired nitric oxide (NO) metabolism characteristic of salt sensitivity. Twenty, otherwise, healthy obese salt-sensitive subjects completed a 12-month program of caloric restriction, aerobic exercise and metformin. Two salt sensitivity tests were performed, that is at baseline and end of program. Lifestyle-metformin treatment decreased weight (9.8+/-0.3 kg), body mass index (3.9+/-0.2 kg/m(2)), waist (11.5+/-0.5 cm), systolic blood pressure (SBP) (8.6+/-0.4 mm Hg), diastolic blood pressure (DBP) (5.5+/-0.4 mm Hg), triglyceride (40+/-5 mg/dl), fasting (8.3+/-1 microIU/ml) and post-load (20+/-4 microIU/ml) insulin levels, and salt sensitivity. Going from a high-sodium ( approximately 300 mmol) to a low-sodium diet ( approximately 30 mmol of sodium/day) lowered SBP/DBP by 14.7+/-1.7/7.4+/ 0.9 mm Hg at baseline and by 8.6+/-1.9/3.2+/-1.2 mm Hg after treatment (P<0.001). More importantly, blood pressure (BP) sensitivity to customary levels of dietary salt ( approximately 150 mmol of sodium/day) was abolished by the lifestyle metformin treatment. Differences in SBP/DBP between usual and low salt averaged 11+/-1/8+/-1 mm Hg before treatment, and 3+/-1/1+/-0.5 mm Hg after treatment (P<0.001). At baseline, NO-metabolite excretion was inhibited during high salt; this impairment was corrected by the lifestyle-metformin treatment. In conclusion, acquired correctable factors play an important role in the pathogenesis of salt sensitivity associated with obesity. Correction of salt sensitivity may account for the BP lowering induced by weight reduction. Restoration of the inability to increase or sustain NO production in response to high salt could account for the correction of salt sensitivity induced by the lifestyle-metformin treatment. PMID- 17460714 TI - Assessment of single versus twice daily dosing of ramipril by ambulatory blood pressure monitoring in patients similar to those included in the HOPE study. AB - Ramipril has been used in twice daily dose of 5 mg in most heart failure trials, whereas the dose used in Heart Outcomes Prevention Evaluation (HOPE) study was 10 mg once at bedtime. The HOPE investigators in an ambulatory blood pressure (ABP) substudy observed a fall of nighttime but not daytime blood pressure (BP). We examined the effects of once daily ramipril 10 mg versus 5 mg twice a day. Twenty nine patients were recruited based on the original criteria for the HOPE study and were given ramipril either in twice-daily dose (5 mg b.d.) or once daily (10 mg o.d.) each morning in randomized, prospective crossover trial. Twenty-four hour ABP recordings were taken just before commencement of ramipril therapy and after treatment with twice-daily and once-daily ramipril. Our results show that ramipril causes a significant reduction of BP over 24-h period as compared with baseline. The mean baseline ABP was 124/73 mm Hg, which reduced to 117/69 mm Hg for the twice-a-day regimen (P<0.001) and 115/68 mm Hg for the once a day regimen (P<0.001). Both regimes effectively lower BP to a similar extent. Ramipril causes significant BP reduction in both once- and twice-daily dosing. The fall in BP after daytime dosing is greater than that observed in the HOPE study (including ABP substudy). Once-daily dosing in the morning seems to be effective in causing a significant reduction in the ABP profile of patients at high-risk of a future vascular event. PMID- 17460715 TI - Highlights of the 23rd Genes and Cancer meeting University of Warwick 11-13 December 2006. PMID- 17460717 TI - Significantly increased lifespan and improved behavioral performances by rAAV gene delivery in adult mucopolysaccharidosis IIIB mice. AB - Mucopolysaccharidosis (MPS) IIIB is an inherited lysosomal storage disease, caused by the deficiency of alpha-N-acetylglucosaminidase (NaGlu), resulting in severe global neurological involvement with high mortality. One major hurdle in therapeutic development for MPS IIIB is the presence of the blood-brain barrier, which impedes the global central nervous system (CNS) delivery of therapeutic materials. In this study, we used a minimal invasive strategy, combining an intravenous (i.v.) and an intracisternal (i.c.) injection, following an i.v. infusion of mannitol, to complement the CNS delivery of adeno-associated viral (AAV) vector for treating MPS IIIB in young adult mice. This treatment resulted in a significantly prolonged lifespan of MPS IIIB mice (11.1-19.5 months), compared with that without treatment (7.9-11.3), and correlated with significantly improved behavioral performances, the restoration of functional NaGlu, and variable correction of lysosomal storage pathology in the CNS, as well as in different somatic tissues. This study demonstrated the great potential of combining i.v. and i.c. administration for improving rAAV CNS gene delivery and developing rAAV gene therapy for treating MPS IIIB in patients. PMID- 17460716 TI - Reversal of streptozotocin-induced diabetes in rats by gene therapy with betacellulin and pancreatic duodenal homeobox-1. AB - Ultrasound-targeted microbubble destruction (UTMD) was used to direct betacellulin (BTC) and pancreatic duodenal homeobox-1 (PDX1) to rat pancreas 48 h after islet destruction by streptozotocin (STZ). Sprague-Dawley rats were rendered diabetic by STZ injection. Controls included normal rats, STZ only without UTMD, and UTMD with DsRed reporter gene. Blood glucose increased dramatically in all rats 48 h after STZ, and continued to rise after UTMD with BTC alone. Blood glucose declined from day 3 to day 10 after UTMD with PDX1, but remained elevated (261+/-8 mg/dl). However, in rats treated with both BTC and PDX1, blood glucose remained below 200 mg/dl throughout day 10. This was accompanied by normalization of blood insulin and C-peptide. Histology demonstrated islet-like clusters of glucagon-staining cells in the rats treated with BTC and PDX1, but these clusters disappeared by 30 days after UTMD treatment. Although regeneration of insulin-producing islets was not seen, diabetes was reversed for up to 15 days after a single UTMD treatment by ectopic insulin production by pancreatic acinar cells. These cells co-expressed amylase and insulin and demonstrated several beta-cell markers by reverse transcription PCR. Gene therapy by UTMD can reverse diabetes in vivo in adult rats by restoring pancreatic insulin production. PMID- 17460718 TI - Integration of active human beta-galactosidase gene (100 kb) into genome using HSV/AAV amplicon vector. AB - Vectors based on herpes simplex virus type-1 (HSV-1) permit delivery of transgenes of up to 150 kb, while the inverted terminal repeats and Rep of the adeno-associated virus (AAV) can confer site-specific integration into the AAVS1 site, which allows sustained expression of a transgene. In this study, combination of the viral elements in HSV/AAV hybrid vectors has been applied for the infectious transfer of the human lysosomal beta-galactosidase (BGAL) gene of 100 kb. Temporary expression and functional activity of beta-galactosidase (beta gal) could be detected in human beta-gal-deficient patient and glioblastoma (Gli36) cells upon infection with the basic BGAL amplicon vector. Sustained expression of beta-gal was achieved in Gli36 cells infected with rep-plus, but not rep-minus, HSV/AAV hybrid vectors. None of five clones isolated after rep minus hybrid vector infection showed elevated beta-gal activity or site-specific integration. In contrast, 80% of the rep-plus clones possessed beta-gal activity at least twofold greater than normal levels for up to 4 months of continuous growth, and 33% of the clones exhibited AAVS1-specific integration of the ITR flanked transgene. One of the rep-plus clones displayed integration of the ITR cassette only at the AAVS1 site, with no sequences outside the cassette detectable and beta-gal activity fourfold above normal levels. These data demonstrate AAVS1-specific integration of an entire genomic locus and expression of the transgene from the endogenous promoter mediated by an HSV/AAV hybrid vector. PMID- 17460719 TI - Delayed administration of adenoviral BMP-2 vector improves the formation of bone in osseous defects. AB - The direct, local, administration of adenovirus carrying human BMP-2 cDNA (Ad.BMP 2) heals critical-sized femoral bone defects in rabbit and rat models. However, the outcome is suboptimal and the technology needs to provide a more reliable and uniform outcome. To this end, we investigated whether the timing of Ad.BMP-2 administration influenced the formation of mineralized tissue within the defect. Critical-sized defects were created in the femora of 28 Sprague-Dawley rats. Animals were injected intralesionally with a single, percutaneous injection of Ad.BMP-2 (4 x 10(8) plaque-forming units) either intraoperatively (day 0) or 24 h (day 1), 5 days or 10 days after surgery. The femora were evaluated 8 weeks after surgery by X-ray, microcomputed tomography, dual-energy X-ray absorptiometry and biomechanical testing. The incidence of radiological union was markedly increased when administration of Ad.BMP-2 was delayed until days 5 and 10, at which point 86% of the defects healed. These time points also provided greater bone mineral content within the defect site and improved the average mechanical strength of the healed bone. Thus, delaying the injection of Ad.BMP-2 until 5 or 10 days after surgery enables a greater percentage of critical-sized, segmental defects to achieve radiological union, producing a repair tissue with enhanced mineralization and greater mechanical strength. PMID- 17460720 TI - Impact of deletion of envelope-related genes of recombinant Sendai viruses on immune responses following pulmonary gene transfer of neonatal mice. AB - We demonstrated previously that the additive-type recombinant Sendai virus (rSeV) is highly efficient for use in pulmonary gene transfer; however, rSeV exhibits inflammatory responses. To overcome this problem, we tested newly developed non transmissible constructs, namely, temperature-sensitive F-deleted vector, rSeV/dF (ts-rSeV/dF) and a rSeV with all the envelope-related genes deleted (rSeV/dFdMdHN), for pulmonary gene transfer in neonatal mice, by assessing their toxicity and immune responses. The gene expression in the lungs of neonatal ICR mice peaked on day 2, then gradually decreased until almost disappearing at 14 days after infection in all constructs. Loss of body weight and mortality rate, however, were dramatically improved in mice treated with SeV/dFdMdHN (mortality=0%, n=41) and ts-rSeV/dF (24.2%, n=33) compared with additive rSeV (70.7%, n=58). Although the deletion of envelope-related genes of SeV had a small impact on the production of antibody and cytotoxic T-lymphocyte activity in both adults and neonates, a dramatic reduction was found in the events related to innate responses, including the production of proinflammatory cytokines, particularly in the case of neonates. These results indicate that pulmonary gene transfer using SeV/dFdMdHN warrants further investigation for its possible use in developing safer therapeutics for neonatal lung diseases, including cystic fibrosis. PMID- 17460721 TI - Biocompatible micellar nanovectors achieve efficient gene transfer to vascular lesions without cytotoxicity and thrombus formation. AB - Gene therapy, a promising treatment for vascular disease, requires appropriate gene vectors with high gene transfer efficiency, good biocompatibility and low cytotoxicity. To satisfy these requirements from the approach of nonviral vectors, a novel block copolymer, poly(ethylene glycol) (PEG)-block-polycation, carrying ethylenediamine units in the side chain (PEG-b-P[Asp(DET)]) was prepared. PEG-b-P[Asp(DET)] formed a polyplex micelle through polyion complex formation with plasmid DNA (pDNA). The PEG-b-P[Asp(DET)] polyplex micelle showed efficient gene expression with low cytotoxicity against vascular smooth muscle cells in vitro. It also showed reduced interactions with blood components, offering its feasibility of gene delivery via the vessel lumen. To evaluate in vivo gene transfer efficiency for vascular lesions, PEG-b-P[Asp(DET)] micelle was instilled into rabbit carotid artery with neointima by an intravascular method, and expression of the reporter gene in vascular lesions was assessed. Polyplexes from homopolymer P[Asp(DET)] and branched polyethyleneimine (BPEI) were used as controls. Ultimately, only the polyplex micelle showed appreciable gene transfer into vascular lesions without any vessel occlusion by thrombus, which was in strong contrast to BPEI and P[Asp(DET)] polyplexes which frequently showed occlusion with thrombus. These findings suggest that the PEG-b-P[Asp(DET)] polyplex micelle may have promising potential as a nonviral vector for the treatment of vascular diseases. PMID- 17460722 TI - Recombinant AAV-mediated VEGF gene therapy induces mandibular condylar growth. AB - Craniofacial anomalies resulting from impaired growth of mandibular condyles require multidisciplinary interventions, which impose a substantial burden on patients and their families. So far, correcting such deformities with an alternative strategy - gene therapy - is still an uncharted territory. Here, we established an effective in vivo gene delivery system with recombinant adeno associated virus (rAAV)-mediated vascular endothelial growth factor (VEGF) to enhance mandibular condylar growth. With in situ hybridization, RT-PCR, immunostaining and Western blot, transgene expression was clearly detected in the mandibular condyles during the whole experiment periods. At defined time points, specific osteogenetic markers (alkaline phosphatase and osteocalcin) and chondrogenetic markers (collagen type II and collagen type X) were assessed by means of biochemical analysis and their expression significantly changed from day 30. Proliferation index by proliferating cell nuclear antigen staining showed also a significant increase in cell proliferation. Morphological measurement identified that the size of mandibular condyle significantly increased from day 30. Taken together, rAAV-VEGF was successfully established as an efficient delivery system to induce mandibular condylar growth, which provides the basis for future gene therapy to treat patients with craniofacial deformities. PMID- 17460724 TI - Heritability of serum hs-CRP concentration and 5-year changes in the Stanislas family study: association with apolipoprotein E alleles. AB - We aimed at estimating additive genetic heritability, household component effect and the influence of common alleles of the apolipoprotein E gene (APOE) on serum high-sensitivity C-reactive protein (hs-CRP) concentrations and the subsequent changes over 5 years. A sub-sample of 320 nuclear families was randomly selected from the Stanislas Family Study. Serum hs-CRP concentration was measured by immunonephelometry at entrance and after 5 years. APOE alleles were determined by restriction fragment length polymorphism. After adjustment for covariates, the number of the epsilon4 allele was negatively associated with serum concentration of hs-CRP in the whole sample, at entrance and 5 years later, without significant interaction with sex by generation groups (P=0.003 and P=0.0003, respectively). However, no significant association was found between epsilon4 allele and 5-year changes in hs-CRP concentration. Using a variance component analysis, no significant genetic influence was shown in family aggregation of both hs-CRP measurements and 5-year changes; the household common component was between 6.5 and 12.8%. In addition, after adjustment for APOE gene polymorphisms, degrees of resemblance were almost unchanged. In the Stanislas Family Study, epsilon4 allele of the APOE gene was associated with lower hs-CRP concentration, but not with 5 year changes. However, variance component analysis did not evidence a significant polygenic effect. PMID- 17460725 TI - A promoter nucleotide variant of the dendritic cell-specific DCNP1 associates with serum IgE levels specific for dust mite allergens among the Korean asthmatics. AB - Dendritic cells (DCs), the most abundant antigen-presenting cells in the lung, have been drawing attention for their roles in specific allergic responses to aeroallergens with support of Th lymphocytes, and in persistent inflammatory changes in allergic asthma. To identify genetic factors that may be involved in the asthma susceptibility and development of the disease phenotypes, we examined association of DC-specific DCNP1 polymorphisms with the disease risk. The case control study revealed association of the nucleotide variants with serum immunoglobulin E (IgE) levels specific for Dermatophagoides farinae (Der f 1) and Dermatophagoides pteronyssinus (Der p 1), major aeroallergens of dust mites, among subjects with asthma. In particular, the T-allele-carrying genotype frequencies for one of the variants (c.-1289C>T) located in the promoter region were found increased in the asthmatic group with low levels of the mite-specific IgE (odds ratio (OR)=0.63 (0.48-0.83) for Der p 1). Results from functional analyses indicated that the promoter variant would affect the gene expression by modulating DNA-protein interaction. We propose that the genetic polymorphism of DCNP1 may influence production of specific IgE by altering DC functions in the mite allergen presenting and/or processing. The functional relevance of the genetic variation would provide an important insight into the genetic basis of allergic response to the mite antigens. PMID- 17460726 TI - The constitutive capacity of human keratinocytes to kill Staphylococcus aureus is dependent on beta-defensin 3. AB - Normal skin is often exposed to bacteria, including potent pathogens such as E. coli, Staphylococcus aureus, and Streptococcus sp., but these microbes usually do not cause skin inflammation or infection in healthy individuals. Therefore, we hypothesized that there must be a constitutive mechanism for rapid destruction and elimination of small numbers of bacteria which penetrate the stratum corneum from everyday activities. This study found that exposure of keratinocytes cultured from a number of individuals to S. aureus resulted in approximately 2-3 log better killing than by HaCaT cells within 1 hour. Killing required contact between the keratinocytes and the bacteria, but was not dependent on internalization. Contact between the bacteria and the keratinocytes resulted in rapid deposition of several antimicrobial peptides onto the bacteria, but only human beta-defensin (HBD) 3 accumulated at levels sufficient to account for killing when S. aureus were exposed to human skin explants. Blocking peptide binding of HBD3 inhibited killing of the bacteria, indicating an essential role for beta-defensin 3 in the constitutive killing of bacteria by normal keratinocytes. PMID- 17460727 TI - Enhanced anti-mycobacterial immunity in children with erythema nodosum and a positive tuberculin skin test. AB - Erythema nodosum (EN) may follow a variety of infections, but in regions with a high prevalence of tuberculosis, is frequently associated with a positive tuberculin skin test (TST) and tuberculosis infection. We aimed to investigate the immunological differences between patients with EN as a manifestation of primary tuberculosis, and those with progressive pulmonary tuberculosis (PTB) or asymptomatic infection. We studied the inflammatory response to both mycobacterial and non-mycobacterial antigens in 11 children with EN associated with a positive TST, 22 children with culture-confirmed tuberculosis, and 53 healthy skin test-positive children. In addition, we evaluated functional anti mycobacterial immunity using an ex vivo assay of mycobacterial growth restriction in five children with EN and 15 with PTB. Patients with EN were distinguished by enhanced mycobacterial growth restriction on the functional assay, which was associated with a markedly increased production of IFNgamma in response to stimulation with purified protein derivative of Mycobacterium tuberculosis. Children presenting with EN and a positive TST show evidence of responses associated with enhanced anti-mycobacterial immunity. PMID- 17460728 TI - Isolation and expression of a Malassezia globosa lipase gene, LIP1. AB - Dandruff and seborrheic dermatitis (D/SD) are common hyperproliferative scalp disorders with a similar etiology. Both result, in part, from metabolic activity of Malassezia globosa and Malassezia restricta, commensal basidiomycete yeasts commonly found on human scalps. Current hypotheses about the mechanism of D/SD include Malassezia-induced fatty acid metabolism, particularly lipase-mediated breakdown of sebaceous lipids and release of irritating free fatty acids. We report that lipase activity was detected in four species of Malassezia, including M. globosa. We isolated lipase activity by washing M. globosa cells. The isolated lipase was active against diolein, but not triolein. In contrast, intact cells showed lipase activity against both substrates, suggesting the presence of at least another lipase. The diglyceride-hydrolyzing lipase was purified from the extract, and much of its sequence was determined by peptide sequencing. The corresponding lipase gene (LIP1) was cloned and sequenced. Confirmation that LIP1 encoded a functional lipase was obtained using a covalent lipase inhibitor. LIP1 was differentially expressed in vitro. Expression was detected on three out of five human scalps, as indicated by reverse transcription-PCR. This is the first step in a molecular description of lipid metabolism on the scalp, ultimately leading toward a test of its role in D/SD etiology. PMID- 17460729 TI - Nox1 induces differentiation resistance in immortalized human keratinocytes generating cells that express simple epithelial keratins. AB - We have shown that superoxide radical-generating NADPH oxidase 1 (Nox1) is increased in intermediate human transformed cells. It was unknown whether Nox1 overexpression could accelerate early transformation steps. We demonstrated that Nox1 rendered human immortalized (GM16) keratinocytes resistant against Ca(2+)/serum-induced differentiation. Nox1-transfected cells produced fast dividing resistant cells within 7-10 days after DMEM exposure. Progenitor lines (or Nox1 lines) were reproducibly generated from Nox1-transfected cells, while no lines were obtained from control transfections. From several attempts to generate control cells, one resistant population was obtained from untransfected GM16 cells after a 6-week DMEM exposure. Prolonged passaging of the control line could induce Nox1. Compared with the control line, Nox1 lines showed greater expression of Nox1, Rac1, p47phox, p67phox, NOXO1, and NOXA1 with concomitant increased superoxide generation. All five Nox1 lines contained varying amounts of E cadherin, involucrin, vimentin, and K8/K18, while the control line did not. Since vimentin and K8/K18 are associated with malignant progression in different types of human epithelial tumors, our data demonstrate that Nox1 accelerated neoplastic like progression by inducing generation of progenitor cells. Our data also emphasize the importance of Nox1 in inducing resistance against differentiation induced cell death, suggesting a contribution of Nox1 and its oxidants during early stage of cell transformation. PMID- 17460730 TI - Systemic anti-TNFalpha treatment restores diabetes-impaired skin repair in ob/ob mice by inactivation of macrophages. AB - To date, diabetes-associated skin ulcerations represent a therapeutic problem of clinical importance. The insulin-resistant type II diabetic phenotype is functionally connected to obesity in rodent models of metabolic syndrome through the release of inflammatory mediators from adipose tissue. Here, we used the impaired wound-healing process in obese/obese (ob/ob) mice to investigate the impact of obesity-mediated systemic inflammation on cutaneous wound-healing processes. Systemic administration of neutralizing monoclonal antibodies against tumor necrosis factor (TNF)alpha (V1q) or monocyte/macrophage-expressed EGF-like module-containing mucin-like hormone receptor-like (Emr)-1 (F4/80) into wounded ob/ob mice at the end of acute wound inflammation initiated a rapid and complete neo-epidermal coverage of impaired wound tissue in the presence of a persisting diabetic phenotype. Wound closure in antibody-treated mice was paralleled by a marked attenuation of wound inflammation. Remarkably, anti-TNFalpha- and anti F4/80-treated mice exhibited a strong reduction in circulating monocytic cells and reduced numbers of viable macrophages at the wound site. Our data provide strong evidence that anti-TNFalpha therapy, widely used in chronic inflammatory diseases in humans, might also exert effects by targeting "activated" TNFalpha expressing macrophage subsets, and that inactivation or depletion of misbehaving macrophages from impaired wounds might be a novel therapeutic clue to improve healing of skin ulcers. PMID- 17460731 TI - Post-transcriptional regulation of melanin biosynthetic enzymes by cAMP and resveratrol in human melanocytes. AB - Upregulation of microphthalmia-associated transcription factor (MITF) expression has been proposed to mediate melanogenesis stimulated by cAMP, whereas downregulation of MITF has been suggested to underlie the depigmentary effects of resveratrol, a promising chemotherapeutic found in red wine. We have assessed the contribution of MITF to pigmentation regulation by treating primary cultures of normal human melanocytes with the adenylate cyclase activator forskolin and/or resveratrol, then quantifying mRNA and protein levels for MITF, tyrosinase, tyrosinase-related protein-1, and dopachrome tautomerase (DCT). The inhibition of tyrosinase activity by resveratrol was not due to alterations in MITF, but instead was explained by both direct tyrosinase inhibition and a post transcriptional effect that reduced the amount of fully processed tyrosinase. Glycosidase digestion revealed that the basis for the tyrosinase decrease was the retention of an immature form in the ER and subsequent loss of the mature, Golgi processed enzyme. Elevation of intracellular cAMP by forskolin markedly increased protein levels for MITF, tyrosinase and DCT, however there was no concomitant increase in tyrosinase or DCT mRNA. This indicated that elevated levels of MITF were not sufficient to promote transcription of these melanogenic genes and that the increase in their protein abundance appeared to be predominantly mediated through post-transcriptional processing events. PMID- 17460732 TI - Improving sun-protection behavior among children: results of a cluster-randomized trial in Italian elementary schools. The "SoleSi SoleNo-GISED" Project. AB - A history of sunburns in early life nearly doubles the risk of developing malignant melanoma in adulthood. From 2001 to 2004, we conducted a cluster randomized trial of an educational intervention to reduce sunburn rates (primary outcome) and improve sun-protection behavior (secondary outcome) in schoolchildren. A total of 122 Italian primary schools (grades 2 and 3) were randomized to receive, or not, an intervention consisting of an educational curriculum at school, conducted by trained teachers, which included the projection of a short video and the distribution of booklets to children and their parents. Behavior while in the sun was assessed at baseline and 14-16 months after baseline. In a subgroup (44% of the total sample), melanocytic nevi were also counted. Of the 11,230 children enrolled, 8,611 completed the study. A total of 1,547 children (14%) reported a history of sunburns at baseline. At follow-up, no difference in sunburn episodes was documented between the study groups (odds ratio 0.97, 95% confidence interval 0.84-1.13) and similar sun protection habits were reported. No significant impact of the proposed educational program was documented at 1-year follow-up. Innovative strategies need to be developed to increase the effectiveness of future educational interventions in this area. PMID- 17460733 TI - Five new homozygous mutations in the KIND1 gene in Kindler syndrome. PMID- 17460734 TI - Noninvasive imaging of melanoma with reflectance mode confocal scanning laser microscopy in a murine model. AB - A reflectance-mode confocal scanning laser microscope (rCSLM) was developed for imaging early-stage melanoma in a living mouse model without the addition of exogenous contrast agents. Lesions were first located by surveying the dorsum with a polarized light camera, then imaged with the rCSLM. The images demonstrated two characteristics of melanoma in this animal model: (1) melanocytes and apparent tumor nests in the epidermis at the stratum spinosum in a state of pagetoid spread and (2) architectural disruption of the dermal epidermal junction. The epidermal melanocytes and apparent tumor nests had a high melanin content, which caused their reflectance to be fivefold greater than the surrounding epidermis. The rCSLM images illustrate the difference between normal skin and sites with apparent melanoma. This imaging modality shows promise to track the progression of melanoma lesions in animal models. PMID- 17460735 TI - Impaired induction of adhesion molecule expression in immortalized endothelial cells leads to functional defects in dynamic interactions with lymphocytes. AB - Immortalization should overcome the problem of short lifespan and difficult culture of endothelial cells that limited their use in functional studies. We used four different immortalized endothelial cell lines to study dynamic interactions with lymphocytes. Surprisingly, tumor necrosis factor (TNF)alpha stimulated human umbilical vein endothelial cells (HUVECs) or human dermal microvascular endothelial cells (HDMECs) readily supported rolling and binding of lymphocytes, whereas none of the immortalized cell lines did. As rolling interactions are primarily mediated by selectins and vascular cell adhesion molecule (VCAM)-1, the endothelial cells were analyzed regarding expression of selectins and other adhesion molecules. Interestingly, cell surface expression of E-selectin could only be detected on HUVEC and HDMEC. Immunocytochemistry showed that some immortalized endothelial cells expressed E-selectin intracellularly following TNFalpha stimulation, suggesting translation but defective post translational processing or transport of the molecule. In contrast, other immortalized cell lines did not have detectable levels of E-selectin mRNA, suggesting impaired transcription. VCAM-1 could only be induced on normal and human placental microvascular endothelial cell-A2 endothelial cells, whereas all cell lines expressed intercellular adhesion molecule-1 following TNF stimulation. The immortalized endothelial cells tested here have lost functions that are required for dynamic interactions with immune cells and that are common to primary endothelial cells. PMID- 17460736 TI - Targeting tumor-associated macrophages and inhibition of MCP-1 reduce angiogenesis and tumor growth in a human melanoma xenograft. AB - Chemokines such as monocyte chemoattractant protein (MCP)-1 are key agonists that attract macrophages to tumors. In melanoma, it has been previously shown that variable levels of MCP-1/CCL2 appear to correlate with infiltrating macrophages and tumor fate, with low to intermediate levels of the chemokine contributing to melanoma development. To work under such conditions, a poorly tumorigenic human melanoma cell line was transfected with an expression vector encoding MCP-1. We found that M2 macrophages are associated to MCP-1+ tumors, triggering a profuse vascular network. To target the protumoral macrophages recruitment and reverting tumor growth promotion, clodronate-laden liposomes (Clod-Lip) or bindarit were administered to melanoma-bearing mice. Macrophage depletion after Clod-Lip treatment induced development of smaller tumors than in untreated mice. Immunohistochemical analysis with an anti-CD31 antibody revealed scarce vascular structures mainly characterized by narrow vascular lights. Pharmacological inhibition of MCP-1 with bindarit also reduced tumor growth and macrophage recruitment, rendering necrotic tumor masses. We suggest that bindarit or Clod Lip abrogates protumoral-associated macrophages in human melanoma xenografts and could be considered as complementary approaches to antiangiogenic therapy. PMID- 17460737 TI - Prevention and control of sexually transmitted infections in New Zealand. PMID- 17460738 TI - The integration of managed healthcare in New Zealand. PMID- 17460739 TI - Chronic Care Management evolves towards Integrated Care in Counties Manukau, New Zealand. AB - Despite anecdotes of many chronic care management and integrated care projects around New Zealand, there is no formal process to collect and share relevant learning within (but especially between) District Health Boards (DHBs). We wish to share our experiences and hope to stimulate a productive exchange of ongoing learning. We define chronic care management and integrated care, then summarise current theory and evidence. We describe national policy development (relevant to integrated care, since 2000) including the New Zealand Health Strategy, the NZ Primary Care Strategy, the development of Primary Health Organisations (PHOs), capitation payments, Care Plus, and Services to Improve Access funding. We then describe chronic care management in Counties Manukau, which evolved both prior to and during the international refinement of theory and evidence and the national policy development and implementation. We reflect on local progress to date and opportunities for (and barriers to) future improvements, aided by comparative reflections on the United Kingdom (UK). Our most important messages are addressed as follows: To policymakers and funders--a fragile culture change towards teamwork in the health system is taking place in New Zealand; this change needs to be specifically and actively supported. To PHOs--general practices need help to align their internal (within-practice) financial signals with the new world of capitation and integrated care. To primary and secondary care doctors, nurses, and other carers - systematic chronic care management and integrated care can improve patient quality of life; and if healthcare structures and systems are properly managed to support integration, then healthcare provider professional and personal satisfaction will improve. PMID- 17460740 TI - Chlamydia screening in Wellington Family Planning Association (FPA) clinics: a demonstration project. AB - AIMS: To demonstrate that enhanced screening for Chlamydia over and above the usual opportunistic screening in family planning (FPA) clinics is feasible, practical, and acceptable. METHODS: Over a 6-month period from November 2004 to May 2005, all under-25-year-olds attending three Wellington FPA clinics in New Zealand were offered Chlamydia urine testing. Staff interviews before and after the study were carried out to assess the impact of enhanced screening on clinic routines. Interviews were conducted with 50 clients to assess the acceptability to young persons. Additional questions were asked of 22 Chlamydia-positive clients to ascertain the acceptability of the procedures for follow up. RESULTS: From a total of 4674 participants, a valid urine test was carried out on 2533 (54%). The most common reason for exclusion was having passed urine in the last hour. Positive tests were detected in 212 (8%). A positive result was more likely in those with a history of partner change or in Maori and Pacific ethnic groups; it was least likely in those who always used condoms. For the staff, time constraints were the most important barrier to screening. The procedures were acceptable to clients. CONCLUSIONS: We demonstrated that improvements in Chlamydia screening are feasible, practical and acceptable to clients PMID- 17460741 TI - Risk factors and characteristics of patients with gonorrhoea presenting to Auckland Sexual Health Service, New Zealand. AB - AIMS: A case-control study of patients with gonorrhoea presenting to Auckland Sexual Health Service was carried out to determine specific risk factors and to look more closely at the transmission of ciprofloxacin-resistant Neisseria gonorrhoeae in the Auckland region. METHODS: Patients who tested positive for gonorrhoea during a 6-month time period from September 2003 till March 2004 were included in the study. A control group was selected from patients who presented during the same time period but who had negative sexually transmitted infection (STI) screens. A selection of N. gonorrhoeae isolates were sent to ESR for typing by pulsed-field gel electrophoresis (PFGE). RESULTS: 204 cases of gonorrhoea were identified during the study period; 50% of the diagnosed cases were in people aged less than 26 years. Gonococcal infection was strongly associated with being of either Maori or Pacific ethnicity (p<0.001). A history of consistent condom use was found to lower the risk of acquisition of gonorrhoea (p=0.009). Thirty three percent of cases had ciprofloxacin-resistant gonococcal infection and the majority of these acquired their infection in New Zealand (88%). Comparison of PFGE genotyping data with results from an audit in 2001 indicated that genetic diversity amongst ciprofloxacin-resistant isolates has increased significantly. CONCLUSION: The incidence of gonorrhoea in the Auckland population is steadily rising, with the main risk factors being age under 25 and being of Maori or Pacific ethnicity. Genetic diversity amongst ciprofloxacin-resistant isolates is increasing, thus indicating high levels of endemic infection. Urgent action needs to be taken to control the current gonorrhoea epidemic in Auckland and more research is required to investigate reasons for ethnic differences in disease incidence. PMID- 17460742 TI - Treatment of anaphylaxis in adults: results of a survey of doctors at Dunedin Hospital, New Zealand. AB - AIMS: To identify which medications doctors would prescribe when treating an adult patient with anaphylaxis, and to ascertain the dose and route of administration of adrenaline they would use. METHOD: Doctors of various grades working in a range of acute specialties at Dunedin Public Hospital (n=91) were asked to anonymously complete a questionnaire detailing two hypothetical cases of anaphylaxis. RESULTS: 92% of participants would give adrenaline as first-line treatment to a patient with anaphylaxis, but only 20% knew the correct dose and route of administration according to the New Zealand Resuscitation Council (NZRC) or local hospital formulary guidelines. 43% of doctors surveyed stated they would give adrenaline by the intravenous (IV) route as first-line treatment with 20% proposing a dose of 1 milligram or greater. CONCLUSION: Most doctors surveyed were not clear about current anaphylaxis treatment guidelines. In particular, they were unsure of the recommended dose and route of administration of adrenaline. To ensure that the first-line treatment of anaphylaxis is safe, we recommend that intramuscular (IM) adrenaline should be used in the majority of situations in accordance with both NZRC and local hospital guidelines. We recommend that all doctors should receive regular education concerning the treatment of anaphylaxis. PMID- 17460743 TI - Health outcomes for diabetes patients returning for three annual general practice checks. AB - AIM: To measure changes in the health status of 840 Type 1 and 9998 Type 2 diabetes patients who had completed three free annual diabetes reviews by December 2005. METHOD: We evaluated changes in clinical measures and differences in proportions of patients achieving guideline targets between the first and third reviews. Logistic regression analysis examined variables associated with an absolute decrease in HbA1c level of at least 1% from the first to the third review. RESULTS: The proportion of patients achieving guideline targets for glycaemic control fell from 17.1% to 12.2% for Type 1 diabetes (p=0.004) and from 56.1% to 50.2% for Type 2 diabetes (p<0.001). There were significant improvements in mean albumin:creatinine ratio, total cholesterol, and high density lipoprotein levels in both diabetes cohorts and mean blood pressure and triglyceride levels in Type 2 patients. Adjusted odds of improved glycaemic control were higher for Type 2 diabetics treated with oral medication only (OR=4.27 (95%CI: 3.45-5.14)), insulin only (OR=7.10 [95%CI: 5.58-9.04]), and insulin and oral therapy (OR=10.05 [95%CI: 7.72-13.09]). CONCLUSION: The introduction of a structured and systematic general practice review process aimed at improving diabetes care and patient outcomes resulted in significant improvements in mean blood pressure, cholesterol levels, and albumin:creatinine ratio. There was, however, no overall improvement in glycaemic control PMID- 17460744 TI - Too much of a good thing, is it bad? Adrenaline on trial. AB - Adrenaline is vital in the treatment of severe allergic reactions (anaphylaxis), however it is often underutilised or inappropriately administered. Adrenaline treatment is not without risk and most adverse reactions to adrenaline occur when it is given in overdose or as an intravenous bolus. We report a case of myocardial injury and hypotension following inappropriate administration of adrenaline. PMID- 17460745 TI - Seek and ye shall find: Hodgkin's lymphoma presenting as paraneoplastic cerebellar degeneration. AB - Paraneoplastic neurological syndromes (PNS) are rare complications of cancer and may be the presenting sign of an occult malignancy. Diagnosis of PNS is often challenging and treatment of the underlying tumour remains the best therapeutic option for preventing further neurological damage. We report on a patient who developed a rapidly progressive cerebellar ataxia and underwent extensive investigations before a paraneoplastic aetiology was confirmed by detection of an underlying malignancy. PMID- 17460746 TI - Disappearing drain--disaster averted and lesson learnt! AB - The use of postoperative drains date back to Hippocrates. We report an iatrogenic case of migrated drain into the retroperitoneum. A novel technique using a rigid cystoscope for retrieval is described that prevented another laparotomy. PMID- 17460747 TI - The prevalence of misleading tobacco descriptors in the New Zealand tobacco market. PMID- 17460748 TI - Medical image. Pneumomediastinum and pneumopericardium. PMID- 17460749 TI - Chlamydia trachomatis: discovery of a new strain. PMID- 17460750 TI - Prevention of colchicine toxicity in patients with gout. PMID- 17460751 TI - Burns treatment--with a response from Middlemore Hospital's Burn Unit. PMID- 17460752 TI - Focusing on fear: Attentional disengagement from emotional faces. AB - Evidence suggests that anxiety is associated with a shift of visual attention toward threatening stimuli in the environment, such as facial expressions (Mogg & Bradley, 1999). More recent evidence, however, indicates that anxiety may be better characterized by a failure to rapidly disengage the visual attention system away from threat-related facial expressions (Fox, Russo, Bowles, & Dutton, 2001). The present study further investigates this delayed disengagement hypothesis. Results show that high trait-anxious individuals, in contrast to low trait-anxious individuals, take longer to classify peripheral target letters when fearful facial expressions were presented at fixation relative to sad, happy, or neutral expressions. These findings demonstrate a specific tendency to dwell on fear-relevant stimuli, as opposed to negative information in general. These findings are considered from an evolutionary perspective and the possible role of delayed disengagement from threat in the maintenance of anxiety states is also discussed. PMID- 17460753 TI - Mood-congruent free recall bias in anxiety. AB - The present study evaluated the status of mood-congruent free recall bias in anxious individuals following incidental encoding of target words. In the first experiment, high trait anxiety individuals showed increased recall of threat related information after an orienting task promoting lexical processing of target words. In a second experiment, both lexical and semantic orienting tasks were performed at study. In this experiment, anxious individuals displayed a mood congruent recall bias only for target information processed at a lexical level. Theoretical and practical implications of these findings are discussed. PMID- 17460754 TI - Identification of the endogenous key substrates of the human organic cation transporter OCT2 and their implication in function of dopaminergic neurons. AB - BACKGROUND: The etiology of neurodegenerative disorders, such as the accelerated loss of dopaminergic neurons in Parkinson's disease, is unclear. Current hypotheses suggest an abnormal function of the neuronal sodium-dependent dopamine transporter DAT to contribute to cell death in the dopaminergic system, but it has not been investigated whether sodium-independent amine transporters are implicated in the pathogenesis of Parkinson's disease. METHODOLOGY/PRINCIPAL FINDINGS: By the use of a novel tandem-mass spectrometry-based substrate search technique, we have shown that the dopaminergic neuromodulators histidyl-proline diketopiperazine (cyclo(his-pro)) and salsolinol were the endogenous key substrates of the sodium-independent organic cation transporter OCT2. Quantitative real-time mRNA expression analysis revealed that OCT2 in contrast to its related transporters was preferentially expressed in the dopaminergic regions of the substantia nigra where it colocalized with DAT and tyrosine hydroxylase. By assessing cell viability with the MTT reduction assay, we found that salsolinol exhibited a selective toxicity toward OCT2-expressing cells that was prevented by cyclo(his-pro). A frequent genetic variant of OCT2 with the amino acid substitution R400C reduced the transport efficiency for the cytoprotective cyclo(his-pro) and thereby increased the susceptibility to salsolinol-induced cell death. CONCLUSIONS/SIGNIFICANCE: Our findings indicate that the OCT2 regulated interplay between cyclo(his-pro) and salsolinol is crucial for nigral cell integrity and that a shift in transport efficiency may impact the risk of Parkinson's disease. PMID- 17460755 TI - Enriched monolayer precursor cell cultures from micro-dissected adult mouse dentate gyrus yield functional granule cell-like neurons. AB - BACKGROUND: Stem cell cultures are key tools of basic and applied research in Regenerative Medicine. In the adult mammalian brain, lifelong neurogenesis originating from local precursor cells occurs in the neurogenic regions of the hippocampal dentate gyrus. Despite widespread interest in adult hippocampal neurogenesis and the use of mouse models to study it, no protocol existed for adult murine long-term precursor cell cultures with hippocampus-specific differentiation potential. METHODOLOGY/PRINCIPAL FINDINGS: We describe a new strategy to obtain serum-free monolayer cultures of neural precursor cells from microdissected dentate gyrus of adult mice. Neurons generated from these adherent hippocampal precursor cell cultures expressed the characteristic markers like transcription factor Prox1 and showed the TTX-sensitive sodium currents of mature granule cells in vivo. Similar to granule cells in vivo, treatment with kainic acid or brain derived neurotrophic factor (BDNF) elicited the expression of GABAergic markers, further supporting the correspondence between the in vitro and in vivo phenotype. When plated as single cells (in individual wells) or at lowest density for two to three consecutive generations, a subset of the cells showed self-renewal and gave rise to cells with properties of neurons, astrocytes and oligodendrocytes. The precursor cell fate was sensitive to culture conditions with their phenotype highly influenced by factors within the media (sonic hedgehog, BMP, LIF) and externally applied growth factors (EGF, FGF2, BDNF, and NT3). CONCLUSIONS/SIGNIFICANCE: We report the conditions required to generate adult murine dentate gyrus precursor cell cultures and to analyze functional properties of precursor cells and their differentiated granule cell-like progeny in vitro. PMID- 17460756 TI - Self-reactivities to the non-erythroid alpha spectrin correlate with cerebral malaria in Gabonese children. AB - BACKGROUND: Hypergammaglobulinemia and polyclonal B-cell activation commonly occur in Plasmodium sp. infections. Some of the antibodies produced recognize self-components and are correlated with disease severity in P. falciparum malaria. However, it is not known whether some self-reactive antibodies produced during P. falciparum infection contribute to the events leading to cerebral malaria (CM). We show here a correlation between self-antibody responses to a human brain protein and high levels of circulating TNF alpha (TNFalpha), with the manifestation of CM in Gabonese children. METHODOLOGY: To study the role of self reactive antibodies associated to the development of P. falciparum cerebral malaria, we used a combination of quantitative immunoblotting and multivariate analysis to analyse correlation between the reactivity of circulating IgG with a human brain protein extract and TNFalpha concentrations in cohorts of uninfected controls (UI) and P. falciparum-infected Gabonese children developing uncomplicated malaria (UM), severe non-cerebral malaria (SNCM), or CM. RESULTS/CONCLUSION: The repertoire of brain antigens recognized by plasma IgGs was more diverse in infected than in UI individuals. Anti-brain reactivity was significantly higher in the CM group than in the UM and SNCM groups. IgG self reactivity to brain antigens was also correlated with plasma IgG levels and age. We found that 90% of CM patients displayed reactivity to a high-molecular mass band containing the spectrin non-erythroid alpha chain. Reactivity with this band was correlated with high TNFalpha concentrations in CM patients. These results strongly suggest that an antibody response to brain antigens induced by P. falciparum infection may be associated with pathogenic mechanisms in patients developing CM. PMID- 17460757 TI - FAK-Mediated mechanotransduction in skeletal regeneration. AB - The majority of cells are equipped to detect and decipher physical stimuli, and then react to these stimuli in a cell type-specific manner. Ultimately, these cellular behaviors are synchronized to produce a tissue response, but how this is achieved remains enigmatic. Here, we investigated the genetic basis for mechanotransduction using the bone marrow as a model system. We found that physical stimuli produced a pattern of principal strain that precisely corresponded to the site-specific expression of sox9 and runx2, two transcription factors required for the commitment of stem cells to a skeletogenic lineage, and the arrangement and orientation of newly deposited type I collagen fibrils. To gain insights into the genetic basis for skeletal mechanotransduction we conditionally inactivated focal adhesion kinase (FAK), an intracellular component of the integrin signaling pathway. By doing so we abolished the mechanically induced osteogenic response and thus identified a critical genetic component of the molecular machinery required for mechanotransduction. Our data provide a new framework in which to consider how physical forces and molecular signals are synchronized during the program of skeletal regeneration. PMID- 17460758 TI - Diversity of raft-like domains in late endosomes. AB - BACKGROUND: Late endosomes, the last sorting station in the endocytic pathway before lysosomes, are pleiomorphic organelles composed of tubular elements as well as vesicular regions with a characteristic multivesicular appearance, which play a crucial role in intracellular trafficking. Here, we have investigated whether, in addition to these morphologically distinguishable regions, late endosomal membranes are additionally sub-compartmentalized into membrane microdomains. METHODOLOGY/PRINCIPAL FINDINGS: Using sub-organellar fractionation techniques, both with and without detergents, combined with electron microscopy, we found that both the limiting membrane of the organel and the intraluminal vesicles contain raft-type membrane domains. Interestingly, these differentially localized domains vary in protein composition and physico-chemical properties. CONCLUSIONS/SIGNIFICANCE: In addition to the multivesicular organization, we find that late endosomes contain cholesterol rich microdomains both on their limiting membrane and their intraluminal vesicles that differ in composition and properties. Implications of these findings for late endosomal functions are discussed. PMID- 17460759 TI - Extracellular sulfatases, elements of the Wnt signaling pathway, positively regulate growth and tumorigenicity of human pancreatic cancer cells. AB - BACKGROUND: Heparan sulfate proteoglycans (HSPGs) are control elements in Wnt signaling, which bind extracellularly to Wnt ligands and regulate their ability to interact with signal transduction receptors on the cell surface. Sulf-1 and Sulf-2 are novel extracellular sulfatases that act on internal glucosamine-6 sulfate (6S) modifications within HSPGs and thereby modulate HSPG interactions with various signaling molecules, including Wnt ligands. Emerging evidence indicates the importance of reactivated Wnt signaling in a number of cancers, including pancreatic adenocarcinoma. PRINCIPLE FINDINGS: Both Sulf proteins were upregulated in human pancreatic adenocarcinoma tumors and were broadly expressed in human pancreatic adenocarcinoma cell lines. Expression of human extracellular sulfatases Sulf-1 and Sulf-2 enhanced Wnt signaling in a reconstituted system. Three of four pancreatic adenocarcinoma cell lines tested exhibited autocrine Wnt signaling, in that extracellular Wnt ligands were required to initiate downstream Wnt signaling. Exposure of these pancreatic adenocarcinoma cells to a catalytically inactive form of Sulf-2 or siRNA-mediated silencing of endogenous Sulf-2 inhibited both Wnt signaling and cell growth. Sulf-2 silencing in two of these lines resulted in markedly reduced tumorigenesis in immunocompromised mice. CONCLUSIONS/SIGNIFICANCE: We have identified the Sulfs as potentiators of autocrine Wnt signaling in pancreatic cancer cells and have demonstrated their contribution to the growth and tumorigenicity of these cells. Since the Sulfs are extracellular enzymes, they would be attractive targets for therapy of pancreatic cancer. Our results run counter to the prevailing view in the literature that the Sulfs are negative regulators of tumorigenesis. PMID- 17460760 TI - Modulation of CD4(+) T cell-dependent specific cytotoxic CD8(+) T cells differentiation and proliferation by the timing of increase in the pathogen load. AB - BACKGROUND: Following infection with viruses, bacteria or protozoan parasites, naive antigen-specific CD8(+) T cells undergo a process of differentiation and proliferation to generate effector cells. Recent evidences suggest that the timing of generation of specific effector CD8(+) T cells varies widely according to different pathogens. We hypothesized that the timing of increase in the pathogen load could be a critical parameter governing this process. METHODOLOGY/PRINCIPAL FINDINGS: Using increasing doses of the protozoan parasite Trypanosoma cruzi to infect C57BL/6 mice, we observed a significant acceleration in the timing of parasitemia without an increase in mouse susceptibility. In contrast, in CD8 deficient mice, we observed an inverse relationship between the parasite inoculum and the timing of death. These results suggest that in normal mice CD8(+) T cells became protective earlier, following the accelerated development of parasitemia. The evaluation of specific cytotoxic responses in vivo to three distinct epitopes revealed that increasing the parasite inoculum hastened the expansion of specific CD8(+) cytotoxic T cells following infection. The differentiation and expansion of T. cruzi-specific CD8(+) cytotoxic T cells is in fact dependent on parasite multiplication, as radiation-attenuated parasites were unable to activate these cells. We also observed that, in contrast to most pathogens, the activation process of T. cruzi-specific CD8(+) cytotoxic T cells was dependent on MHC class II restricted CD4(+) T cells. CONCLUSIONS/SIGNIFICANCE: Our results are compatible with our initial hypothesis that the timing of increase in the pathogen load can be a critical parameter governing the kinetics of CD4(+) T cell-dependent expansion of pathogen-specific CD8(+) cytotoxic T cells. PMID- 17460761 TI - The origin of phenotypic heterogeneity in a clonal cell population in vitro. AB - BACKGROUND: The spontaneous emergence of phenotypic heterogeneity in clonal populations of mammalian cells in vitro is a rule rather than an exception. We consider two simple, mutually non-exclusive models that explain the generation of diverse cell types in a homogeneous population. In the first model, the phenotypic switch is the consequence of extrinsic factors. Initially identical cells may become different because they encounter different local environments that induce adaptive responses. According to the second model, the phenotypic switch is intrinsic to the cells that may occur even in homogeneous environments. PRINCIPAL FINDINGS: We have investigated the "extrinsic" and the "intrinsic" mechanisms using computer simulations and experimentation. First, we simulated in silico the emergence of two cell types in a clonal cell population using a multiagent model. Both mechanisms produced stable phenotypic heterogeneity, but the distribution of the cell types was different. The "intrinsic" model predicted an even distribution of the rare phenotype cells, while in the "extrinsic" model these cells formed small clusters. The key predictions of the two models were confronted with the results obtained experimentally using a myogenic cell line. CONCLUSIONS: The observations emphasize the importance of the "ecological" context and suggest that, consistently with the "extrinsic" model, local stochastic interactions between phenotypically identical cells play a key role in the initiation of phenotypic switch. Nevertheless, the "intrinsic" model also shows some other aspects of reality: The phenotypic switch is not triggered exclusively by the local environmental variations, but also depends to some extent on the phenotypic intrinsic robustness of the cells. PMID- 17460762 TI - Magnetoreception system in honeybees (Apis mellifera). AB - Honeybees (Apis mellifera) undergo iron biomineralization, providing the basis for magnetoreception. We showed earlier the presence of superparamagnetic magnetite in iron granules formed in honeybees, and subscribed to the notion that external magnetic fields may cause expansion or contraction of the superparamagnetic particles in an orientation-specific manner, relaying the signal via cytoskeleton (Hsu and Li 1994). In this study, we established a size density purification procedure, with which quantitative amount of iron granules was obtained from honey bee trophocytes and characterized; the density of iron granules was determined to be 1.25 g/cm(3). While we confirmed the presence of superparamagnetic magnetite in the iron granules, we observed changes in the size of the magnetic granules in the trophycytes upon applying additional magnetic field to the cells. A concomitant release of calcium ion was observed by confocal microscope. This size fluctuation triggered the increase of intracellular Ca(+2) , which was inhibited by colchicines and latrunculin B, known to be blockers for microtubule and microfilament syntheses, respectively. The associated cytoskeleton may thus relay the magnetosignal, initiating a neural response. A model for the mechanism of magnetoreception in honeybees is proposed, which may be applicable to most, if not all, magnetotactic organisms. PMID- 17460763 TI - Searching for tissue-specific expression pattern-linked nucleotides of UGT1A isoforms. AB - UDP-glucuronosyltransferases 1A isoforms belong to a superfamily of microsomal enzymes responsible for glucuronidation of numerous endogenous and exogenous compounds. The nine functional UGT1A isoforms are encoded by a single UGT1A gene locus with multiple first exons. The expression of the UGT1A transcripts was measured by quantitative RT-PCR in 23 normal human tissues. The tissue-specific expression patterns were observed in 13 tissues. To understand the regulation mechanism that is responsible for the tissue-specific expression patterns, we scanned the DNA sequence alignments of the putative promoter regions, exon 1 sequences and intron 1 sequences for those expression-pattern-linked nucleotides. Using one of the expression-pattern-linked nucleotides for livers as an example, we showed that a database comprised of these expression-pattern-linked nucleotides could be used to generate focused hypotheses on the problem of tissue specific expression, which is critical for tissue-specific pharmacodynamics of anticancer drugs. PMID- 17460764 TI - Dynamic social adaptation of motion-related neurons in primate parietal cortex. AB - Social brain function, which allows us to adapt our behavior to social context, is poorly understood at the single-cell level due largely to technical limitations. But the questions involved are vital: How do neurons recognize and modulate their activity in response to social context? To probe the mechanisms involved, we developed a novel recording technique, called multi-dimensional recording, and applied it simultaneously in the left parietal cortices of two monkeys while they shared a common social space. When the monkeys sat near each other but did not interact, each monkey's parietal activity showed robust response preference to action by his own right arm and almost no response to action by the other's arm. But the preference was broken if social conflict emerged between the monkeys-specifically, if both were able to reach for the same food item placed on the table between them. Under these circumstances, parietal neurons started to show complex combinatorial responses to motion of self and other. Parietal cortex adapted its response properties in the social context by discarding and recruiting different neural populations. Our results suggest that parietal neurons can recognize social events in the environment linked with current social context and form part of a larger social brain network. PMID- 17460765 TI - Tbx2 and Tbx3 regulate the dynamics of cell proliferation during heart remodeling. AB - BACKGROUND: The heart forms from a linear tube that is subject to complex remodeling during embryonic development. Hallmarks of this remodeling are the looping of the heart tube and the regionalization into chamber and non-chamber myocardium. Cardiomyocytes in the future chamber myocardium acquire different cellular and physiological characteristics through activation of a chamber specific genetic program, which is in part mediated by T-box genes. METHODOLOGY/PRINCIPAL FINDING: We characterize two new zebrafish T-box transcription factors, tbx3b and tbx2a, and analyze their role during the development of the atrioventricular canal. Loss- and gain-of-function analyses demonstrate that tbx3b and tbx2a are necessary to repress the chamber-genetic program in the non-chamber myocardium. We also show that tbx3b and tbx2a are required to control cell proliferation in the atrioventricular canal and that misregulation of cell proliferation in the heart tube influences looping. Furthermore, we characterize the heart phenotype of a novel Tbx3 mutation in mice and show that both the control of cell proliferation and the repression of chamber-specific genetic program in the non-chamber myocardium are conserved roles of Tbx3 in this species. CONCLUSIONS/SIGNIFICANCE: Taken together, our results uncover an evolutionarily conserved role of Tbx2/3 transcription factors during remodeling of the heart myocardium and highlight the importance of controlling cell proliferation as a driving force of morphogenesis. PMID- 17460766 TI - Comparison of FGF1 (aFGF) expression between the dorsal motor nucleus of vagus and the hypoglossal nucleus of rat. AB - Neurons in the dorsal motor nucleus of the vagus (DMNV) are more severely affected by axonal injury than most other nerves, such as those of the hypoglossal nucleus. However, the mechanism underlying such a response remains unclear. In this study, we compared the expression of fibroblast growth factor 1 (FGF1), a neurotrophic factor, between the DMNV and the hypoglossal nucleus by RT PCR and immunohistochemical analyses. RT-PCR showed that the level of FGF1 mRNA expression in the DMNV was lower than that in the hypoglossal nucleus (P<0.01). Immunohistochemistry revealed that FGF1 was localized to neurons. FGF1-positive neurons in large numbers were evenly distributed in the hypoglossal nucleus, whereas FGF1-positive neurons were located in the lateral part of the DMNV. Double immunostaining for FGF1 and choline acetyltransferase demonstrated that 22.7% and 78% of cholinergic neurons were positive for FGF1 in the DMNV and hypoglossal nucleus, respectively. A tracing study with cholera toxin B subunit (CTb) demonstrated that cholinergic neurons sending their axons from the DMNV to the superior laryngeal nerve were FGF1-negative. The results suggest that the low expression of FGF1 in the DMNV is due to severe damage of neurons in the DMNV. PMID- 17460767 TI - Alterations in somatostatin cells and biochemical parameters following zinc supplementation in gastrointestinal tissue of streptozotocin-induced diabetic rats. AB - Chronic hyperglycemia in diabetes is a major causative factor of free radical generation which further leads to many secondary diabetic complications via the damage to cellular proteins, membrane lipids, and nucleic acids. Zinc is an essential trace element in all living systems and plays a structural role in many proteins and enzymes. Somatostatin is known to have inhibitory effects on various gastrointestinal functions. Therefore, we determined somatostatin protein production and secretion levels, and biochemical and light microscopical changes following zinc supplementation in the gastrointestinal tract of streptozotocin (STZ)-diabetic rats. The animals were divided into four groups: Group I: control (untreated) animals; Group II: control animals given zinc sulfate; Group III: diabetic animals; and Group IV: diabetic animals given zinc sulfate. Zinc sulfate was given to the animals by gavage at a daily dose of 100 mg/kg body weight for 60 days. Diabetes was induced by intraperitoneal (i.p.) injection of STZ in a single dose of 65 mg/kg. For histological studies, stomach and duodenum tissues were fixed in Bouin solution and sections stained with Masson's trichrome and Periodic-Acid-Schiff. Tissue homogenates were used for protein, lipid peroxidation (LPO), glutathione (GSH), and nonenzymatic glycosylation (NEG) analyses. Zinc supplementation to the STZ-diabetic rats revealed the protective effect of zinc on these parameters. Zinc supplementation may contribute to prevent at least some complications of diabetes mellitus. PMID- 17460768 TI - Interleukin-8 producing malignant fibrous histiocytoma with prolonged fever. AB - We present a case of malignant fibrous histiocytoma accompanied by prolonged spiking fevers, which disappeared after tumor resection. Sarcoma with fever as a primary symptom is rare. Furthermore, in this case, fever was closely related to the clinical course of the tumor. In order to detect possible production of febriferous substance(s), we used blood and tumor tissue samples to investigate nine candidate cytokines possibly responsible for the fever. Expression of IL-8 mRNA was detected in preoperative peripheral blood mononuclear cells by RT-PCR. Expressions of IL-6, IL-8, IFN-gamma and TNF-alpha mRNAs were also detected in tumor tissue, while IL-1alpha, IL-1beta, IL-2, IL-4 and COX-2 mRNAs were not. We suspected IL-8 to be a causative factor, and examined its localization by immunohistochemical staining, paraffin sections of tumor tissue stained positive for IL-8. Since infiltrating mononuclear cells were positive for IL-8, this may explain the tumor-associated fever. This case involves intratumoral production of IL-8 as a causative factor, and IL-6, IL-8, IFN-gamma and TNF-alpha cytokine production might have resulted from stimulation with a substance(s) derived from tumor tissue, since the fever disappeared postoperatively. To date the patient is alive and in good health for 7 years and 2 months since the surgery. PMID- 17460769 TI - Clinicopathological characteristics of esophageal squamous papillomas in Japanese patients--with comparison of findings from Western countries. AB - To clarify the characteristics of esophageal squamous papillomas (ESPs) in the Japanese population, we investigated 38 ESPs of 35 Japanese patients from a file with 17,387 upper gastrointestinal endoscopies in our university hospital. ESPs accounted for 0.20% of the total number of endoscopies and comprised 21 females and 14 males with an average age of 59.2 years. More than half of the ESPs (52.6%) were located in the middle esophagus. The ratio of human papilloma virus (HPV) positive ESPs was 10.5% and all were located in the middle esophagus of female patients only. HPV-positive ESP cases were younger (46.8 years) than HPV negative cases (60.8 years). Based on comparison with the reports from western countries, we attribute the low prevalence in the lower esophagus to the relatively fewer occurrences of severe reflux esophagitis (RE) due to chronic gastritis with low gastric acid secretion among Japanese patients. PMID- 17460770 TI - Telomere length on chromosome 17q shortens more than global telomere length in the development of breast cancer. AB - It is known that total telomere length is shorter in invasive breast cancer than in normal breast tissue but the status of individual telomere lengths has not been studied. Part of the difficulty is that usually telomere length in interphase cells is measured on all chromosomes together. In this study we compared normal breast epithelium, duct carcinoma in situ (DCIS), and invasive duct carcinoma (IDC) from 18 patients. Telomere length was specifically measured on chromosome 17q and was found to be shorter in DCIS and IDC than in normal breast epithelial cells, with more heterogeneity in telomere length in DCIS associated with IDC than in DCIS alone. More importantly, we found that the shortening of telomere on chromosome 17q is greater than the average shortening of all telomeres. This finding indicates that telomere shortening is not simply the result of the end replication problem; otherwise, all telomeres should be subjected to the same rate of telomere shortening. It seems there are mechanisms that preferentially erode some telomeres more than others or preferentially protect some chromosome ends. Our results suggest that the increased level of telomere shortening on 17q may be involved in chromosome instability and the progression of DCIS. PMID- 17460771 TI - The HIV protease inhibitor nelfinavir downregulates Akt phosphorylation by inhibiting proteasomal activity and inducing the unfolded protein response. AB - HIV protease inhibitors (HPIs), which have been used to treat HIV patients since the mid 1990s, have been shown to downregulate the phosphatidylinositol 3-kinase (PI3K)-Akt pathway. Because this pathway is frequently activated in human malignancies and associated with resistance to ionizing radiation, we investigated and confirmed that HPIs could radiosensitize cells. However, the mechanism underlying this downregulation was unclear, prompting the investigations in this report. In this paper we show that nelfinavir inhibits proteasome activity. Inhibition of the proteasome leads to endoplasmic reticulum based stress with accumulation of misfolded proteins, which triggers the unfolded protein response (UPR). As part of the UPR, the alpha subunit of eukaryotic translation initiation factor 2 (eIF2alpha) is phosphorylated, resulting in a decrease in global protein synthesis and induction of the feedback regulator growth arrest and DNA damage-inducible protein (GADD34), which acts as a phosphatase in complex with protein phosphatase 1. This complex dephosphorylates eIF2alpha; however, our data also suggest that this phosphatase activity can dephosphorylate Akt. Furthermore, our data indicate that nelfinavir decreases Akt phosphorylation by triggering this response. These findings may have important implications in understanding how nelfinavir may increase radiation sensitivity and also result in downregulation of the PI3K/Akt pathway. PMID- 17460772 TI - Signal transducer and activator of transcription 3 activation promotes invasive growth of colon carcinomas through matrix metalloproteinase induction. AB - Signal transducer and activator of transcription 3 (STAT3) is aberrantly activated in colorectal carcinomas (CRCs). Here, we define the relationship between STAT3 function and the malignant properties of colon carcinoma cells. Elevated activation of STAT3 enhances invasive growth of the CRC cell lines. To address mechanisms through which STAT3 influences invasiveness, the protease mRNA expression pattern of CRC biopsies was analyzed and correlated with the STAT3 activity status. These studies revealed a striking coincidence of STAT3 activation and strong expression of matrix metalloproteinases MMP-1, -3, -7, and 9. Immunohistological examination of CRC tumor specimens showed a clear colocalization of MMP-1 and activated STAT3. Experimentally induced STAT3 activity in CRC cell lines enhanced both the level of MMP-1 mRNA and secreted MMP 1 enzymatic activity. A direct connection of STAT3 activity and transcription from the MMP-1 promoter was shown by reporter gene experiments. Moreover, high affinity binding of STAT3 to STAT recognition elements in both the MMP-1 and MMP 3 promoter was demonstrated. Xenograft tumors arising from implantation of CRC cells into nude mice showed simultaneous appearance and colocalization of p-Y STAT3 and MMP-1 expression. Our results link aberrant activity of STAT3 in CRC to malignant tumor progression through upregulated expression of MMPs. PMID- 17460773 TI - Feature selection and molecular classification of cancer using genetic programming. AB - Despite important advances in microarray-based molecular classification of tumors, its application in clinical settings remains formidable. This is in part due to the limitation of current analysis programs in discovering robust biomarkers and developing classifiers with a practical set of genes. Genetic programming (GP) is a type of machine learning technique that uses evolutionary algorithm to simulate natural selection as well as population dynamics, hence leading to simple and comprehensible classifiers. Here we applied GP to cancer expression profiling data to select feature genes and build molecular classifiers by mathematical integration of these genes. Analysis of thousands of GP classifiers generated for a prostate cancer data set revealed repetitive use of a set of highly discriminative feature genes, many of which are known to be disease associated. GP classifiers often comprise five or less genes and successfully predict cancer types and subtypes. More importantly, GP classifiers generated in one study are able to predict samples from an independent study, which may have used different microarray platforms. In addition, GP yielded classification accuracy better than or similar to conventional classification methods. Furthermore, the mathematical expression of GP classifiers provides insights into relationships between classifier genes. Taken together, our results demonstrate that GP may be valuable for generating effective classifiers containing a practical set of genes for diagnostic/prognostic cancer classification. PMID- 17460775 TI - Cre-loxP-mediated recombination between the SIL and SCL genes leads to a block in T-cell development at the CD4- CD8- to CD4+ CD8+ transition. AB - In the most common form of stem cell leukemia (SCL) gene rearrangement, an interstitial deletion of 82 kb brings SCL under the control of regulatory elements that normally govern expression of the ubiquitously expressed SCL interrupting locus (SIL) gene, which is located directly upstream of SCL. To investigate the effect of this fusion in a mouse model, a bacterial artificial chromosome (BAC) clone containing both human SIL and SCL genes was isolated, and loxP sites were inserted into intron 1 of both the SIL and SCL genes, corresponding to the sites at which recombination occurs in human T-cell acute lymphocytic leukemia patients. This BAC clone was used to generate transgenic SILloxloxSCL mice. These transgenic mice were subsequently bred to Lck-Cre mice that express the Cre recombinase specifically in the thymus. The BAC transgene was recombined between the two loxP sites in over 50% of the thymocytes from SILloxloxSCL/Cre double-transgenic mice, bringing the SCL gene under the direct control of SIL regulatory elements. Aberrant SCL gene expression in the thymus was verified by reverse transcription-polymerase chain reaction. Using FACS analysis, we found that mice carrying both SILloxloxSCL and Cre transgenes have increased CD4-/CD8- thymocytes compared with transgene-negative mice. In the spleen, these transgenic mice show a marked reduction in the number of mature CD4+ or CD8+ cells. These results demonstrate that conditional activation of SCL under control of SIL regulatory elements can impair normal T-cell development. PMID- 17460776 TI - Plant-derived MINA-05 inhibits human prostate cancer proliferation in vitro and lymph node spread in vivo. AB - Few treatment options exist for metastatic prostate cancer (PC) that becomes hormone refractory (HRPC). In vitro, plant-derived MINA-05 caused dose-dependent decreases in cell numbers in HRPC cell lines LNCaP-C4-2B and PC-3, and in androgen-sensitive LNCaP-FGC, DuCaP, and LAPC-4, by WST-1 assay. MINA-05 pretreatment significantly decreased clonogenic survival in agar and on plastic at 1 x and 2 x IC50 for PC-3 (P < .05 and P < .001, respectively), and at 1/2 x, 1 x, and 2 x IC50 for LNCaP-FGC cells (P < .001). MINA-05 also induced G2M arrest of LNCaP-FGC and PC-3 cells (by flow cytometry) and caused some apoptosis in LNCaP-FGC (sub-G1 peak on flow, expression of activated caspase-3) but not in PC 3 cells. Western blotting indicated that these cell cycle changes were associated with decreased levels of regulatory proteins cyclin B1 and cdc25C. MINA-05 given daily by gavage for 39 days did not diminish primary orthotopic PC-3 growth in nude mice, but decreased the extent of lymph node invasion at higher doses. We conclude that MINA-05 induces G2M arrest, inhibits cell growth, reduces PC cell regrowth in vitro, and reduces lymph node invasion after orthotopic PC-3 cell implantation in vivo. It has potential as an adjuvant treatment for patients with PC. PMID- 17460774 TI - Claudin-4 overexpression in epithelial ovarian cancer is associated with hypomethylation and is a potential target for modulation of tight junction barrier function using a C-terminal fragment of Clostridium perfringens enterotoxin. AB - BACKGROUND: Claudin-4, a tight junction (TJ) protein and receptor for the C terminal fragment of Clostridium perfringens enterotoxin (C-CPE), is overexpressed in epithelial ovarian cancer (EOC). Previous research suggests DNA methylation is a mechanism for claudin-4 overexpression in cancer and that C-CPE acts as an absorption-enhancing agent in claudin-4-expressing cells. We sought to correlate claudin-4 overexpression in EOC with clinical outcomes and TJ barrier function, investigate DNA methylation as a mechanism for overexpression, and evaluate the effect of C-CPE on the TJ. METHODS: Claudin-4 expression in EOC was quantified and correlated with clinical outcomes. Claudin-4 methylation status was determined, and claudin-4-negative cell lines were treated with a demethylating agent. Electric cell-substrate impedance sensing was used to calculate junctional (paracellular) resistance (Rb) in EOC cells after claudin-4 silencing and after C-CPE treatment. RESULTS: Claudin-4 overexpression in EOC does not correlate with survival or other clinical endpoints and is associated with hypomethylation. Claudin-4 overexpression correlates with Rb and C-CPE treatment of EOC cells significantly decreased Rb in a dose- and claudin-4 dependent noncytotoxic manner. CONCLUSIONS: C-CPE treatment of EOC cells leads to altered TJ function. Further research is needed to determine the potential clinical applications of C-CPE in EOC drug delivery strategies. PMID- 17460777 TI - Necrosis induction in glioblastoma cells reveals a new "bioswitch" function for the MT1-MMP/G6PT signaling axis in proMMP-2 activation versus cell death decision. AB - Cytoskeleton disorganization is an early step in the activation process of matrix metalloproteinase 2 (MMP-2) by membrane type 1 MMP (MT1-MMP) but is also associated with endoplasmic reticulum (ER) dysfunction and subsequent cell death. Given evidence that the ER-embedded glucose-6-phosphate transporter (G6PT) regulates glioblastoma cell survival and that MT1-MMP is a key enzyme in the cancer cell invasive phenotype, we explored the molecular link between G6PT and MT1-MMP. Cytoskeleton-disrupting agents such as concanavalin A (ConA) and cytochalasin D triggered proMMP-2 activation and cell death in U87 glioma cells. ConA decreased G6PT gene expression, an event that was also observed in cells overexpressing the full-length recombinant MT1-MMP protein. Overexpression of a membrane-bound catalytically active but cytoplasmic domain-deleted MT1-MMP was unable to downregulate G6PT gene expression or to trigger necrosis. Gene silencing of MT1-MMP with small interfering RNA prevented proMMP-2 activation and induced G6PT gene expression. ConA inhibited Akt phosphorylation, whereas overexpression of recombinant G6PT rescued the cells from ConA-induced proMMP-2 activation and increased Akt phosphorylation. Altogether, new functions of MT1 MMP in cell death signaling may be linked to those of G6PT. Our study indicates a molecular signaling axis regulating the invasive phenotype of brain tumor cells and highlights a new "bioswitch" function for G6PT in cell survival. PMID- 17460778 TI - Oncogenic K-Ras signals through epidermal growth factor receptor and wild-type H Ras to promote radiation survival in pancreatic and colorectal carcinoma cells. AB - Pancreatic and colorectal carcinomas frequently express oncogenic/mutant K-Ras that contributes to both tumorigenesis and clinically observed resistance to radiation treatment. We have previously shown that farnesyltransferase inhibitors (FTI) radiosensitize many pancreatic and colorectal cancer cell lines that express oncogenic K-ras at doses that inhibit the prenylation and activation of H Ras but not K-Ras. In the present study, we have examined the mechanism of FTI mediated radiosensitization in cell lines that express oncogenic K-Ras and found that wild-type H-Ras is a contributor to radiation survival in tumor cells that express oncogenic K-Ras. In these experiments, inhibiting the expression of oncogenic K-Ras, wild-type H-Ras, or epidermal growth factor receptor (EGFR) led to similar levels of radiosensitization as treatment with the FTI tipifarnib. Treatment with the EGFR inhibitor gefitinib led to similar levels of radiosensitization, and the combinations of tipifarnib or gefitinib plus inhibition of K-Ras, H-Ras, or EGFR expression did not provide additional radiosensitization compared with tipifarnib or gefitinib alone. Finally, supplementing culture medium with the EGFR ligand transforming growth factor alpha was able to reverse the radiosensitizing effect of inhibiting K-ras expression. Taken together, these findings suggest that EGFR-activated H-Ras signaling is initiated by oncogenic K-Ras to promote radiation survival in pancreatic and colorectal cancers. PMID- 17460779 TI - Tumor vesicle-associated CD147 modulates the angiogenic capability of endothelial cells. AB - Matrix metalloproteinase (MMP) degradation of extracellular matrix is thought to play an important role in invasion, angiogenesis, tumor growth, and metastasis. Several studies have demonstrated that CD147/extracellular MMP inducer, a membrane-spanning molecule highly expressed in tumor cells, may be involved in the progression of malignancies by regulating expression of MMP in peritumoral stromal cells. In the present study we show that CD147 is expressed in microvesicles derived from epithelial ovarian cancer cells and that CD147 positive vesicles may promote an angiogenic phenotype in endothelial cells in vitro. Vesicles shed by human ovarian carcinoma cell lines OVCAR3, SKOV3, and A2780 expressed different levels of CD147 and stimulated proangiogenic activities of human umbilical vein endothelial cells (HUVECs) in a CD147-dependent fashion (OVCAR3 > SKOV3 > A2780). Moreover, vesicles shed by ovarian carcinoma cell line CABA I with low CD147 expression had no significant effect on the development of angiogenic phenotype in HUVECs. The treatment of OVCAR3 cells with small interfering RNA against CD147 suppressed the angiogenic potential of OVCAR3 derived microvesicles. However, transfection of CD147 cDNA into the CABA I cell line enabled CABA I-derived vesicles to induce angiogenesis and to promote MMP genes expression in HUVECs. We therefore conclude that vesicles shed by ovarian cancer cells may induce proangiogenic activities of HUVECs by a CD147-mediated mechanism. PMID- 17460780 TI - Disrupted axo-glial junctions result in accumulation of abnormal mitochondria at nodes of ranvier. AB - Mitochondria and other membranous organelles are frequently enriched in the nodes and paranodes of peripheral myelinated axons, particularly those of large caliber. The physiologic role(s) of this organelle enrichment and the rheologic factors that regulate it are not well understood. Previous studies suggest that axonal transport of organelles across the nodal/paranodal region is locally regulated. In this study, we have examined the ultrastructure of myelinated axons in the sciatic nerves of mice deficient in the contactin-associated protein (Caspr), an integral junctional component. These mice, which lack the normal septate-like junctions that promote attachment of the glial (paranodal) loops to the axon, contain aberrant mitochondria in their nodal/paranodal regions. These mitochondria are typically large and swollen and occupy prominent varicosities of the nodal axolemma. In contrast, mitochondria located outside the nodal/paranodal regions of the myelinated axons appear normal. These findings suggest that paranodal junctions regulate mitochondrial transport and function in the axoplasm of the nodal/paranodal region of myelinated axons of peripheral nerves. They further implicate the paranodal junctions in playing a role, either directly or indirectly, in the local regulation of energy metabolism in the nodal region. PMID- 17460781 TI - 3D dosimetry by optical-CT scanning. AB - The need for an accurate, practical, low-cost 3D dosimetry system is becoming ever more critical as modern dose delivery techniques increase in complexity and sophistication. A recent report from the Radiological Physics Center (RPC) (1), revealed that 38% of institutions failed the head-and-neck IMRT phantom credentialing test at the first attempt. This was despite generous passing criteria (within 7% dose-difference or 4mm distance-to-agreement) evaluated at a half-dozen points and a single axial plane. The question that arises from this disturbing finding is - what percentage of institutions would have failed if a comprehensive 3D measurement had been feasible, rather than measurements restricted to the central film-plane and TLD points? This question can only be adequately answered by a comprehensive 3D-dosimetry system, which presents a compelling argument for its development as a clinically viable low cost dosimetry solution. Optical-CT dosimetry is perhaps the closest system to providing such a comprehensive solution. In this article, we review the origins and recent developments of optical-CT dosimetry systems. The principle focus is on first generation systems known to have highest accuracy but longer scan times. PMID- 17460782 TI - IMRT verification using a radiochromic/optical-CT dosimetry system. PMID- 17460783 TI - Towards four dimensional (4D) dosimetry for radiation-therapy. PMID- 17460784 TI - Entecavir: a new nucleoside analogue for the treatment of chronic hepatitis B. AB - Infection with hepatitis B virus (HBV) is extremely widespread - it infects two billion people out of the six billion world population. It is estimated that between 350 and 400 million people are chronically infected with HBV. Chronic HBV infection leads to development of complications, such as cirrhosis and hepatocellular carcinoma (HCC), which arise in 15-40% of patients. HBV-related liver disease and its complications result in approximately one million deaths each year. The ultimate goals of chronic hepatitis B (CHB) therapy are decreases in the incidence of cirrhosis, end-stage liver disease and HCC. The following six medications are currently approved by the U.S. Food and Drug Administration for the treatment of CHB: interferon (INF)-alpha2b, pegylated INF-alpha2a, lamivudine, adefovir dipivoxil, entecavir and, recently, telbivudine. Interferon therapy has many contraindications and commonly causes multiple intolerable adverse effects. Lamivudine therapy leads to increased development of resistant mutations with each year of use. Entecavir, a new guanosine nucleoside analogue with specific activity against HBV DNA polymerase, represents a third agent within the nucleoside/nucleotide HBV polymerase inhibitor class. It has distinct advantages over lamivudine and adefovir dipivoxil: it has a three-step mechanism of action, is the most potent inhibitor of HBV DNA polymerase, is not associated with any major adverse effects and has a limited potential for resistance. In clinical trials, entecavir was superior to lamivudine in all primary endpoints in both nucleoside-naive and lamivudine-refractory hepatitis B e antigen (HBeAg) positive and HBeAg-negative patients. Preliminary data support entecavir efficacy in patients with cirrhosis and HIV/HBV coinfected patients. No resistance occurred after two years of entecavir therapy in nucleoside-naive patients. Up to 9% resistance developed in patients with documented prior lamivudine resistance during 96 weeks of entecavir therapy. Currently, entecavir should be considered a first- or second-line treatment option for the management of HBeAg-positive or negative nucleoside-naive or lamivudine-refractory CHB patients. PMID- 17460785 TI - Lopinavir/ritonavir: appraisal of its use in HIV therapy. AB - Recommendations for a highly active antiretroviral therapy in either pretreated patients or symptomatic patients with an AIDS-defining event include at least one protease inhibitor. The majority of currently available protease inhibitors are coadministrated with low-dose ritonavir, a pharmacoenhancer that significantly increases protease inhibitor plasma concentrations. In the class of protease inhibitors lopinavir plus ritonavir is the only coformulation. This coformulation was designed to overcome the problems of earlier agents of this class of drugs concerning unfavorable pharmacokinetics with a higher frequency of dosing and therapy failure. The pharmacoenhancing effect of ritonavir on lopinavir resulted in a highly potent, clinically effective antiretroviral drug with a high genetic barrier to viral resistance. Safety concerns have taken a backseat, focusing instead on the favorable efficacy of lopinavir, which recently led to the evaluation of its use in boosted double-protease-inhibitor regimens, as a once daily application and even in HIV monotherapy. Nevertheless, since HIV infection became a chronic but controllable disease, side effects like metabolic disorders and cardiovascular disease have begun to draw increased attention in the long term treatment with protease inhibitors. Coformulated lopinavir/ritonavir is available as a soft gelatin capsule (133.33/33.33 mg), liquid formulation (80/20 mg/ml) and recently approved melt-extrusion tablet (200/50 mg). Lopinavir/ritonavir is recommended for first- and second-line therapy in HIV-1 infection, in children as well as adolescents and adults. PMID- 17460786 TI - Extended-release paliperidone: efficacy, safety and tolerability profile of a new atypical antipsychotic. AB - Extended-release paliperidone is a new atypical antipsychotic chemically related to the well-known antipsychotic risperidone. It has been formulated in an osmotic controlled-release oral delivery system that minimizes peak-trough fluctuations and, by obviating dose-titration, allows once-daily dosing with a therapeutically active dose from the first day. Its pharmacokinetic profile is characterized by a mean time-to-peak plasma concentration of 24.1 hours and an elimination half-life of approximately 24 hours. A dose of 6 mg of paliperidone extended-release (ER) provides a mean striatal D2 receptor occupancy of 64%, approaching the accepted lower receptor occupancy threshold required for optimal antipsychotic activity without causing extrapyramidal symptoms. It undergoes minimal hepatic biotransformation and is mainly excreted unchanged in the urine with four metabolic products. Three pivotal randomized, double-blind, placebo-controlled, parallel-group six-week trials investigated the efficacy, safety and tolerability of paliperidone ER at doses of 3-15 mg/day. All doses produced a significant reduction in schizophrenia symptomatology, with an onset of effect as of day 4. Personal and social functioning also improved as measured by the Personal and Social Performance scale. A prevention of recurrence study showed that paliperidone ER effectively prolonged the time-to-recurrence versus placebo. Paliperidone ER was efficacious in young, elderly and recently diagnosed schizophrenia patients. Beneficial effects on sleep assessed objectively and subjectively with minimal daytime somnolence were demonstrated. Overall, it was well-tolerated and had placebo-like discontinuation rates for adverse events. There were some dose-related extrapyramidal symptoms, mostly mild to moderate in intensity and associated with minimal changes in the rating scales that assessed extrapyramidal symptom severity. Although prolactin elevation occurred with paliperidone ER, only few prolactin-related adverse events were reported. There were no signals for metabolic dysfunction in terms of glucose, insulin, lipid or triglyceride changes or any indicators of clinically relevant cardiac events. Body weight increased slightly, but the changes were acceptable in the context of the doses likely to be used clinically, and patients with higher initial body mass index gained less weight as has been reported for other antipsychotics. PMID- 17460787 TI - Strategies for suppressing angiogenesis in gynecological cancers. AB - Angiogenesis is vital for numerous physiological and pathological processes, including proliferation, invasion and metastasis in malignancies. Various strategies to suppress angiogenesis are under evaluation in gynecological malignancies, and ovarian cancer--the deadliest of them--has been the focus. Interruption of interaction between vascular endothelial growth factor (VEGF) and VEGF receptor (VEGFR) with a monoclonal antibody, bevacizumab, has so far been the most promising antiangiogenic strategy in ovarian cancer clinically but is overshadowed by higher than expected frequency of severe toxicities. Interception of VEGF with receptor decoys, such as VEGF-Trap, and inhibiting receptor tyrosine kinases for VEGF and related growth factors with small molecule inhibitors have shown encouraging results in early phase trials of ovarian cancer; validation is ongoing in larger studies. Another approach is targeting pre-mRNA for VEGF receptors with ribozyme (angiozyme). The knowledge gained from developing these different classes of antiangiogenic agents will lay the path to future trials of other types of gynecological cancers. PMID- 17460789 TI - Visualizing RNA splicing in vivo. AB - Ribozymes are RNA molecules capable of associating with other RNA molecules through base-pairing and catalyzing various reactions involving phosphate group transfer. Of particular interest to us is the well known ribozyme from Tetrahymena thermophila capable of catalyzing RNA splicing in eukaryotic systems, chiefly because of its potential use as a gene therapy agent. In this article we review the progress made towards visualizing the RNA splicing mediated by the Tetrahymena ribozyme in single living mammalian cells with the beta-lactamase reporter system and highlight the development made in imaging RNA splicing with the luciferase reporter system in living animals. PMID- 17460790 TI - Proteomic approaches to studying protein tyrosine phosphatases. AB - Protein tyrosine phosphatases (PTPs) constitute a large family of enzymes that play key roles in cell signaling. Deregulation of PTP activity results in aberrant tyrosine phosphorylation, which has been linked to the etiology of several human diseases, including cancer. Since phosphate removal by the PTPs can both enhance and antagonize cellular signaling, it is essential to elucidate the physiological context in which PTPs operate. Two powerful proteomic approaches have been developed to rapidly establish the exact functional roles for every PTP, both in normal cellular physiology and in pathogenic conditions. In the first, an affinity-based substrate-trapping approach has been employed for PTP substrate identification. Identification and characterization of specific PTP substrate interactions will associate functions with PTP as well as implicate PTP to specific signaling pathways. In the second, a number of activity-based PTP probes have been developed that can provide a direct readout of the functional state of the PTPs in complex proteomes. The ability to profile the entire PTP family on the basis of changes in their activity is expected to yield new functional insights into pathways regulated by the PTPs and contribute to the discovery of PTPs as novel therapeutic targets. Effective application of these proteomic techniques will accelerate the functional characterization of PTPs, thereby facilitating our understanding of PTPs in cell signaling and in diseases. PMID- 17460791 TI - Mammalian antimicrobial proteins and peptides: overview on the RNase A superfamily members involved in innate host defence. AB - The review starts with a general outlook of the main mechanisms of action of antimicrobial proteins and peptides, with the final aim of understanding the biological function of antimicrobial RNases, and identifying the key events that account for their selective properties. Although most antibacterial proteins and peptides do display a wide-range spectrum of action, with a cytotoxic activity against bacteria, fungi, eukaryotic parasites and viruses, we have only focused on their bactericidal activity. We start with a detailed description of the main distinctive structural features of the bacteria target and on the polypeptides, which act as selective host defence weapons.Following, we include an overview of all the current available information on the mammalian RNases which display an antimicrobial activity. There is a wealth of information on the structural, catalytic mechanism and evolutionary relationships of the RNase A superfamily. The bovine pancreatic RNase A (RNase A), the reference member of the mammalian RNase family, has been the main research object of several Nobel laureates in the 60s, 70s and 80s. A potential antimicrobial function was only recently suggested for several members of this family. In fact, the recent evolutionary studies indicate that this protein family may have started off with a host defence function. Antimicrobial RNases constitute an interesting example of proteins involved in the mammalian innate immune defence system. Besides, there is wealth of available information on the mechanism of action of short antimicrobial peptides, but little is known on larger polypeptides, that is, on proteins. Therefore, the identification of the mechanisms of action of antimicrobial RNases would contribute to the understanding of the proteins involved in the innate immunity. PMID- 17460792 TI - Analytical nanobiotechnology for medicine diagnostics. AB - The review is concerned with the state-of-the-art and the prospects of development of nanotechnologies in clinical proteomics. Nanotechnology in clinical proteomics is a new medical research direction, dealing with the creation and application of nanodevices for performing proteomic analyses in the clinic. Nanotechnological progress in the field of atomic force microscopy makes it possible to perform clinical studies on the revelation, visualization and identification of protein disease markers, in particular of those with the sensitivity of 10(-17) M that surpasses by several orders the sensitivity of commonly adopted clinical methods. At the same time, implementation of nanotechnological approaches into diagnostics allows for the creation of new diagnostic systems based on the optical, electro-optical, electromechanical and electrochemical nanosensoric elements with high operating speed. The application of nanotechnological approaches to creating nanopore-based devices for express sequencing of the genome is discussed. PMID- 17460793 TI - A synthetic biology challenge: making cells compute. AB - Advances in biology and engineering have enabled the reprogramming of cells with well-defined functions, leading to the emergence of synthetic biology. Early successes in this nascent field suggest its potential to impact diverse areas. Here, we examine the feasibility of engineering circuits for cell-based computation. We illustrate the basic concepts by describing the mapping of several computational problems to engineered gene circuits. Revolving around these examples and past studies, we discuss technologies and computational methods available to design, test, and optimize gene circuits. We conclude with discussion of challenges involved in a typical design cycle, as well as those specific to cellular computation. PMID- 17460794 TI - Quantitative shotgun proteomics using a protease with broad specificity and normalized spectral abundance factors. AB - Non-specific proteases are rarely used in quantitative shotgun proteomics due to potentially high false discovery rates. Yet, there are instances when application of a non-specific protease is desirable to obtain sufficient sequence coverage of otherwise poorly accessible proteins or structural domains. Using the non specific protease, proteinase K, we analyzed Saccharomyces cerevisiae preparations grown in (14)N rich media and (15)N minimal media and obtained relative quantitation from the dataset using normalized spectral abundance factors (NSAFs). A critical step in using a spectral counting based approach for quantitative proteomics is ensuring the inclusion of high quality spectra in the dataset. One way to do this is to minimize the false discovery rate, which can be accomplished by applying different filters to a searched dataset. Natural log transformation of proteinase K derived NSAF values followed a normal distribution and allowed for statistical analysis by the t-test. Using this approach, we generated a dataset of 719 unique proteins found in each of the three independent biological replicates, of which 84 showed a statistically significant difference in expression levels between the two growth conditions. PMID- 17460795 TI - More systematic reviews needed to improve occupational health. PMID- 17460796 TI - Systematic review of the prevention incentives of insurance and regulatory mechanisms for occupational health and safety. AB - OBJECTIVES: The objective of this study was to determine the strength of evidence on the effectiveness of two policy levers-the experience rating of workers' compensation insurance and the enforcement of occupational health and safety regulation-in creating incentives for firms to focus on health and safety issues. METHODS: An extensive systematic literature review was undertaken in an effort to capture both published and grey literature studies on the topic. Studies that met specific subject-matter and methods criteria underwent a quality assessment. A qualitative approach to evidence synthesis, known as "best-evidence" synthesis, was used. This method ranks the strength of evidence on a particular topic on the basis of the number, quality, and consistency of studies on the topic. RESULTS: There was moderate evidence that the degree of experience rating reduces injuries, limited to mixed evidence that inspections offer general and specific deterrence and that citations and penalties aid general deterrence, and strong evidence that actual citations and penalties reduce injuries. CONCLUSIONS: Although experience rating is a key policy lever of those providing workers' compensation insurance, there is much to be learned about its merits. Few studies have concerned the topic, and most have used crude proxy measures or exploited natural experiments. There have been many more studies on the merits of regulation enforcement, even though here too measures were often crude. Nonetheless, this synthesis indicates that general deterrence is less effective in reducing injury incidence and severity, whereas specific deterrence with regard to citations and penalties does indeed have an impact. PMID- 17460797 TI - Immigrant populations, work and health--a systematic literature review. AB - OBJECTIVES: This paper summarizes the information on immigrant occupational health available from recent studies, incorporating varied study designs. METHODS: A systematic search was carried out in PubMed employing terms of interest to the study and related terms supplied by the same search engine. Articles were selected through the following process: (i) reading the title and abstract, in English or Spanish, for the period 1990-2005, (ii) reading of the entire text of selected articles; (iii) making a manual search of the relevant citations in the selected articles; (iv) eliminating articles without a focus on the themes of central interest (immigration, work, and health), and (v) reading and analyzing the definitive article set. No quality criteria were used in the article selection. RESULTS: The location of studies was not straightforward and required careful thought about the search terms. The included 48 papers were often multifaceted and difficult to categorize. They generally came from countries historically associated with immigration and described occupational risk factors, health consequences, and the social, economic, and cultural influences on worker health. The authors of the studies highlighted the fact that the data were limited and that the [corrected] surveillance, training, and preventive measures used among these populations [corrected] were inadequate. CONCLUSIONS: Increased migration is a reality in industrialized countries all over the world, and it has social, political, and economic consequences for migrating groups, as well as for their sending and host societies. More reliable data, targeted appropriate interventions, and enforcement of existing regulations are necessary to improve the health of immigrant workers. Furthermore, studies in sending and developing countries should be encouraged to form a more complete understanding of this complex situation. PMID- 17460798 TI - Health effects of transitions in work schedule, workhours and overtime in a prospective cohort study. AB - OBJECTIVES: The aim of this study was to examine the longitudinal relationship between transitions in work schedules, workhours and overtime and changes in several self-reported health outcomes (general health, fatigue, need for recovery, and psychological distress). METHODS: Three-year follow-up data from the Maastricht Cohort Study on fatigue at work were used. Gender-stratified logistic regression analyses using generalized estimating equations were conducted for each of the dichotomized health outcomes, with control for a range of possible confounding factors. RESULTS: In this study, transitions in worktime arrangements were prospectively related to changes in several self-reported health outcomes. Substantial and significant associations were found for transitions in work schedule and the incidence of prolonged fatigue and for the need for recovery among men. Moreover, transitions in workhours affected the need for recovery among men, while they influenced general health and psychological distress among women. Finally, transitions in overtime were significantly associated with the incidence of the need for recovery among both men and women and with the incidence of psychological distress among men only. CONCLUSIONS: Transitions in worktime arrangements are related to changes in health, and studying transitions might be an important means of gaining insight into a possible causal relationship between employment and health. Given the considerable impact of worktime arrangements on the individual worker, employers, and society and the high frequency in which transitions within worktime arrangements can occur, these findings underline the need for interventions addressing worktime arrangements in order to reduce or prevent their impact on employee health. PMID- 17460799 TI - Exposure, lung function decline and systemic inflammatory response in asphalt workers. AB - OBJECTIVES: The aim of this study was to determine the association between exposures in asphalt work and changes in lung function, blood concentrations of interleukin-6 (IL-6), micro-C-reactive protein, and fibrinogen among asphalt workers during a work season. METHODS: Blood samples from all asphalt workers (N=140) in Norway's largest road construction and maintenance company were taken in April-May 2005 and again in September-October 2005. Spirometric tests of the asphalt workers and a reference group (heavy construction workers, N=126) were carried out before the asphalt season, and the asphalt workers were tested again at the end of the season. Exposure to total dust, oil mist, polycyclic aromatic hydrocarbons, and gases was measured by personal samplers during the asphalt season. RESULTS: The asphalt workers had a significantly a lower forced expiratory volume in 1 second (FEV(1)) and forced expiratory flow rate of 50% of the forced vital capacity than the reference group at the beginning of the season. The asphalt workers were divided according to their exposure into two groups, asphalt pavers (N=81) and asphalt plant operators and truck drivers (N=54). The screedmen, a group of the asphalt pavers, had a statistically significant lower FVC and FEV(1) after one season of asphalt work than all of the other asphalt workers (P<0.05). The mean plasma concentration of IL-6 increased among the asphalt pavers from 1.55 pg/ml before the season to 2.67 pg/ml at the season's end (P=0.04, adjusted for current smoking). CONCLUSIONS: Exposure in asphalt paving may enhance the risk of lung function decline. PMID- 17460800 TI - Consensus-based findings and recommendations for estimating the costs of health related productivity loss from a company's perspective. AB - OBJECTIVES: There were two study objectives: (i) to identify, via consensus, a key set of items for estimating the costs of productivity loss from a company's perspective and (ii) to develop recommendations for the costs of estimating productivity loss on the basis of consensus findings. METHODS: A modified Delphi procedure was utilized in which a predetermined set of 26 items formed the basis for inquiry in the first round. Thirty-six experts from five stakeholder groups in the Netherlands (employers, employees, policy makers and insurers, occupational health professionals, and researchers) participated in the panel. Opinions were sought regarding the relevance and retrievability of data on items related to the following three forms of work loss: work presenteeism (ie, decreased work performance while at work), short-term absenteeism (<2 weeks), and long-term absenteeism (>2 weeks). The data were analyzed quantitatively and qualitatively. The consensus for relevance was set at 70%. RESULTS: After two rounds, 4 items were found relevant for estimating the costs of productivity loss due to work presenteeism, 6 items were relevant for short-term absenteeism, and 11 items remained for long-term absenteeism. The retrievability of data varied. Three sets of recommendations were formulated for estimating the costs of productivity loss from a company's perspective. CONCLUSION: A streamlined set of relevant items has been identified via consensus and formulated into recommendations for estimating the costs of productivity loss from a company's perspective. Although not definitive, these recommendations represent an important step towards standardizing the way these costs are estimated, and, in turn, facilitate the comparability and utility of economic evaluations of occupational health interventions. PMID- 17460801 TI - A pattern recognition approach to the development of a classification system for upper-limb musculoskeletal disorders of workers. AB - OBJECTIVES: Workers' musculoskeletal disorders are often pain-based and elude specific diagnoses; yet diagnosis or classification is the cornerstone to researching and managing these disorders. Clinicians are skilled in pattern recognition and use it in their daily practice. The purpose of this study was to use the clinical reasoning of experienced clinicians to recognize patterns of signs and symptoms and thus create a classification system. METHODS: Two hundred and forty-two workers consented to a standardized physical assessment and to completing a questionnaire. Each physical assessment finding was dichotomized (normal versus abnormal), and the results were graphically displayed on body diagrams. At two different workshops, groups of experienced researchers or clinicians were led through an exercise of pattern recognition (clustering and naming of clusters) to arrive at a classification system. Interobserver reliability was assessed (8 observers, 40 workers), and the classification system was revised to improve reliability. RESULTS: The initial classification system had good face validity but low interobserver reliability (kappa <0.3). Revisions were made that resulted in a proposed triaxial classification system. The signs and symptoms axes quantified the areas in the involved upper limbs. The proposed third axis described the likelihood of a specific clinical diagnosis being made and the degree of certainty. The interobserver reliability improved to approximately 0.70. CONCLUSIONS: This triaxial classification system for musculoskeletal disorders is based on clinically observable findings. Further testing and application in other populations is required. This classification system could be useful for both clinicians and epidemiologists. PMID- 17460802 TI - Operating room nursing and lung cancer risk in a cohort of female registered nurses. AB - OBJECTIVES: Smoke generated during laser surgery and electrocautery contains respiratory irritants and human carcinogens. Although laboratory and animal studies have demonstrated that this smoke has inflammatory and mutagenic potential, no population-based studies of the health effects of exposure to surgical smoke have been published. We examined the association between duration of employment as an operating room nurse, a proxy measure for surgical smoke exposure, and subsequent lung cancer risk. METHODS: This study was conducted among 86 747 women in the Nurses' Health Study. Information on the duration of prior operating room employment was collected in 1984, and the women were followed for incident, confirmed lung cancer. Cox proportional hazards regression was used to model the incidence rate ratio of lung cancer for each exposure category using women with no prior operating room employment for comparison. All of the models were adjusted for age, smoking history, passive smoke exposure, fruit and vegetable consumption, and alpha carotene and lycopene intake. RESULTS: A history of operating room employment was not associated with an increased rate of lung cancer in multivariable analyses [rate ratio (RR) 0.99, 95% confidence interval (95% CI) 0.86-1.15]. In fact, nurses in the highest exposure category, > or =15 years of operating room employment, had a significantly lower rate of lung cancer than nurses with no prior operating room employment (RR 0.58, 95% CI 0.37 0.91), possibly due to confounding by overall health status or residual confounding by smoking history. CONCLUSIONS: Long-term exposure to surgical smoke, as measured by the duration of operating room employment, does not appear to increase the risk of lung cancer. PMID- 17460803 TI - Electroencephalography artifacts in workplace alertness monitoring. AB - OBJECTIVE: This study assessed the effect of removing artifacts from workplace electroencephalography (EEG) recordings on power spectra and the consequent interpretation of changes in alertness. METHODS: EEG was recorded for 27 air traffic controllers on the night shifts of four roster cycles. On two of the four night shifts, each controller was given a 40-minute opportunity to nap, while on the other two they remained awake (105 shifts in total). Recordings for the last hour of each night shift were screened for artifacts by an experienced viewer (who viewed the EEG in isolation from other electrophysiological recordings). The effects of the nap opportunity on the EEG power spectra were then analyzed in a mixed model analysis of variance in the presence and absence of artifact contaminated data. RESULTS: Overall, 89.3% of the EEG recordings contained artifacts. Removal of these data markedly altered the interpretation of how the nap opportunities affected the EEG power spectra. The spectral parameters of the artifact appeared to be different when the participants were given the opportunity to nap. CONCLUSIONS: Removal of artifacts can dramatically affect the interpretation of workplace EEG recordings. This potential source of error is often unreported. PMID- 17460804 TI - Occupational vocal cord dysfunction due to exposure to wood dust and xerographic toner. AB - OBJECTIVES: Vocal cord dysfunction is a poorly understood entity that is often misdiagnosed as asthma. Both irritant and non-irritant vocal cord dysfunction have been described. This report presents two cases of irritant vocal cord dysfunction secondary to specific environmental exposure, the first to iroko and western red cedar wood (a carpenter) and the second to xerographic printing toner (a secretary). METHODS: Several tests were performed, including chest radiographs, measurements of total serum immunoglobulin E, skin prick tests with common pneumoallergens (as well as iroko and western red cedar in the first case), pulmonary function studies, methacholine challenge testing, specific inhalation challenge performed with suspected agents in a single-blinded fashion, and peak expiratory flow testing and fiberoptic rhinolaryngoscopy (in case 1). RESULTS: During the specific inhalation challenge, the patients showed dysphonia, chest tightness, inspiratory stridor, and flattening of the inspiratory limb of the maximum flow-volume loop in spirometry, with no significant decreases in the level of forced expiratory volume in 1 second; fiberoptic rhinolaryngoscopy confirmed the diagnosis of vocal cord dysfunction in case 1. CONCLUSIONS: It is important to know that agents that can cause occupational asthma can also cause vocal cord dysfunction. The mechanisms by which these agents produce vocal cord dysfunction are unknown. The differences in the clinical presentation of the patients described relative to the reported cases suggest that more than one pathophysiological mechanism may be implicated in the genesis of this entity. PMID- 17460805 TI - Back extensor strength and lumbar spinal mobility are predictors of quality of life in patients with postmenopausal osteoporosis. AB - To assess the effect of multiple factors on quality of life (QOL) in osteoporosis, relationships between the QOL and possible spinal factors were analyzed in 174 postmenopausal women with osteoporosis. Back extensor strength and lumbar spinal mobility were the most important factors for QOL in these patients. INTRODUCTION: Quality of life (QOL) in patients with osteoporosis and vertebral fractures is impaired by the decline of total spinal mobility, although it is not clear to what extent. This study aimed to assess the effect of multiple factors on QOL in patients with osteoporosis. METHODS: QOL of 174 postmenopausal women with osteoporosis (mean, 68 years old) was evaluated using the Japanese Osteoporosis QOL Questionnaire (JOQOL). Correlations between the JOQOL score, bone mineral density (BMD) of the lumbar spine/proximal femur/whole body, the kyphosis angle and mobility of thoracic and lumbar spine, the number of vertebral fractures, grip strengths of dominant and non-dominant hands, and isometric back extensor strength (BES) were analyzed. RESULTS: JOQOL showed significant correlation (p < 0.05) with age (r = -0.303), BES (r = 0.455), dominant and non dominant grip strengths (r = 0.273 and r = 0.255, respectively), number of vertebral fractures (r = -0.282), BMDs of proximal femur and whole body (r = 0.200 and r = 0.157, respectively), lumbar kyphosis angle (r = -0.296), and lumbar spinal mobility (r = 0.345). Multiple regression analysis revealed that the BES and lumbar spinal mobility were the significant contributors to the JOQOL (p < 0.05). CONCLUSION: BES and lumbar spinal mobility are the important factors for QOL in patients with postmenopausal osteoporosis. PMID- 17460806 TI - The mitochondrial DNA control region of Muscidae flies: evolution and structural conservation in a dipteran context. AB - The structure and evolution of the mtDNA control region (CR) and its flanking genes in economically important dipterans from the family Muscidae (Brachycera: Calyptratae), Haematobia irritans, Musca domestica, Atherigona orientalis, and Stomoxys calcitrans are presented in this paper, along with the description of short noncoding intergenic regions possibly related to CR flanking sequences in Stomoxys calcitrans and Ophyra aenescens mtDNAs (ScIR and OaIR, respectively). S. calcitrans showed a large CR with an approximately 550-bp element tandemly repeated and a duplicated tRNA(Ile) gene. The characterization of H. irritans, M. domestica, A. orientalis, and S. calcitrans CR sequences led to the identification of seven conserved sequence blocks homologous to the elements previously described for Calliphoridae and Oestridae species (Brachycera: Calyptratae). Comparative analysis with Drosophila species (Brachycera: Acalyptratae) revealed four conserved regions. The putative functional roles of the conserved elements in the regulation of replication and transcription processes are addressed. The characterization of the structural organization of the mitochondrial genome CR demonstrates the plasticity of the mtDNA molecule in family Muscidae. PMID- 17460807 TI - Protein polymorphism is negatively correlated with conservation of intronic sequences and complexity of expression patterns in Drosophila melanogaster. AB - We report a significant negative correlation between nonsynonymous polymorphism and intron length in Drosophila melanogaster. This correlation is similar to that between protein divergence and intron length previously reported in Drosophila. We show that the relationship can be explained by the content of conserved noncoding sequences (CNS) within introns. In addition, genes with a high regulatory complexity and many genetic interactions also exhibit larger amounts of CNS within their introns and lower values of nonsynonymous polymorphism. The present study provides relevant evidence on the importance of intron content and expression patterns on the levels of coding polymorphism. PMID- 17460808 TI - Phylogenetic analyses under secondary structure-specific substitution models outperform traditional approaches: case studies with diploblast LSU. AB - Many rDNA molecular phylogenetic studies result in trees that are incongruent to either alternative gene tree reconstructions and/or morphological assumptions. One reason for this outcome might be the application of suboptimal phylogenetic substitution models. While the most commonly implemented models describe the evolution of independently evolving characters fairly well, they do not account for character dependencies such as rRNA strands that form a helix in the ribosome. Such nonindependent sites require the use of models that take into account the coevolution of the complete nucleotide pair (doublet). We analyzed 28S rDNA (LSU) demosponge phylogenies using a "doublet" model for pairing sites (rRNA-helices) and compared our findings with the results of "standard" approaches using Bayes factors. We demonstrate that paired and unpaired sites of the same gene result in different reconstructions and that usage of a doublet model leads to more reliable demosponge trees. We show the influence of more sophisticated models on phylogenetic reconstructions of early-branching metazoans and the phylogenetic relationships of demosponge orders. PMID- 17460810 TI - A new technique of hepatic segmentectomy by selective portal venous occlusion using a balloon catheter through a branch of the superior mesenteric vein. AB - BACKGROUND: We describe a new technique of segment-based liver resection by inserting a 6 French balloon catheter into the portal vein via a branch of the superior mesenteric vein. MATERIALS AND METHODS: The catheter was guided into the relevant portal branch supplying the liver segment that contained the tumor. Injection of methylene blue through the catheter delineated the liver segment(s) to be resected. This operation was carried out on 48 patients with a 0% in hospital mortality. RESULTS: The 5-year overall survival was 52.1% and the 5-year disease-free survival was 20.8%. CONCLUSIONS: Our experience showed this technique to be a viable alternative for anatomical liver segmentectomy. PMID- 17460809 TI - Evolution of ITS ribosomal RNA secondary structures in fungal and algal symbionts of selected species of Cladonia sect. Cladonia (Cladoniaceae, Ascomycotina). AB - Evolutionary studies in lichen associations follow that of the fungal symbiont (mycobiont), which is the symbiont after which the lichen is named and forms the majority of the thallus. However, evolution of the algal partner (photobiont) is important to maintain compatibility between symbionts and to optimize productivity of the lichen association. The internal transcribed spacer (ITS) regions of the nuclear ribosomal DNA (rDNA) were examined for primary DNA sequence patterns and for patterns in the secondary structure of the rRNA transcripts in both symbionts of the genus Cladonia. Fungal and algal symbionts show opposite trends in rates of evolution and fragment lengths. Both symbionts showed stronger conservation of the ITS2 structure than the ITS1 structure. Homology was evident in the secondary structures between the two highly divergent chlorophyte and ascomycete taxonomic groups. Most fungal species and all species complexes were polyphyletic. The ITS rDNA of the natural lichen algae from Manitoba and four known algal species is highly similar. The natural lichen algae segregate into highly supported clades by environmental features, suggesting that algae that are already adapted to the environment may associate with germinating fungal propagules in the genus Cladonia. Fungal plasticity may allow the mycobiont to adapt to the environment of the photobiont producing variation in lichen morphology. This might explain the incongruence of phylogenetic patterns between the algal and fungal partners tested and the polyphyly of the fungal species. PMID- 17460811 TI - Is sentinel lymph node mapping in colorectal cancer a future prognostic factor? A meta-analysis. AB - The diagnostic value of sentinel lymph node mapping (SLNM) in patients with colorectal cancer (CRC) is controversial. Prognostic factors for CRC must be detected to improve its treatment. A PubMed query (key words: colorectal cancer, sentinel node) provided 182 studies on the sentinel lymph node (SLN) for CRC, the abstracts of which were reviewed. Altogether, 48 studies dealing with the diagnostic value of SLNM were selected from PubMed, and 6 other studies were retrieved from reviews. We compared the diagnostic value of SLNM with that of conventional histopathologic examination. We used the diagnostic accuracy odds ratio (DAOR) method. Because of significant heterogeneity, we chose the random effect model (Der Simonian and Laird). Statistics were performed on 33 studies, including 1794 patients (1201 colon and 332 rectum cancers). The mean SLNM failure rate was 10%. The global sensitivity and specificity of the SLNM were, respectively, 70% and 81%. The pooled DAOR was 10.7 (95% confidence interval 7.0 16.5). That means that a patient whose SLN is invaded has 10.7 times more risk to be node-positive than an SLN-negative patient. Lymphatic mapping appears to be readily applicable to CRC. One of the main reasons for the heterogeneity is the performance of the SLNM by Saha et al., whose data had better sensitivity (90%) than those in other studies. The SLNM technique should be better standardized in future studies. Understanding the cause of false-negative SLNs (9%) is a major issue to resolve before routinely using this technique in CRC management. The prognostic implication of micrometastases found in SLNs requires further evaluation. PMID- 17460812 TI - Effect of parathyroidectomy on quality of life and neuropsychological symptoms in primary hyperparathyroidism. AB - Neuropsychological symptoms are found in a certain number of patients with primary hyperparathyroidism (PHPT). Preoperative and postoperative quality of life, anxiety, and depression are measured to analyze the impact of parathyroidectomy on these symptoms. In this prospective study, 66 patients underwent parathyroidectomy for PHPT and were evaluated pre- and postoperatively with two validated psychometric instruments (HADS, PHQ-9). Health-related quality of life was measured with a 12-item short-form health survey (SF-12). Preoperatively, the median physical component score (SF-12) of 43.0 and mental component score of 43.5 were lower than those of the general population (52.8 and 54.2 points, respectively). One year postoperatively the mental component score increased to 48.6 (p = 0.011), whereas the physical functioning scale scored 45.3 and therefore did not change significantly (p = 0.585). Preoperatively, symptoms of depression were found in 23.4% of the patients, and 15.6% of the patients displayed symptoms of anxiety (HADS). The prevalence of depression was significantly higher in patients with preoperative serum calcium levels > 11.2 mg/dl (2.8 mmol/L) (p = 0.015). Twelve months postoperatively, the overall proportion of patients with anxiety and depression decreased to 7.8% and 15.7%, respectively (p = NS). The severity of depression as measured with the PHQ-9 declined postoperatively as well. In this study, preoperative neuropsychological symptoms were related to the serum calcium levels. Postoperative health-related quality of life improved significantly. Among patients with preoperative symptoms of depression and anxiety, both symptoms were alleviated significantly at the 12 month follow-up. Therefore, surgery for PHPT seems to be effective in reducing neuropsychological morbidity associated with PHPT. PMID- 17460813 TI - 3D kinematics of the glenohumeral joint during abduction motion: an ex vivo study. AB - The clinical tolerance of rotator cuff tears is extremely variable, so the question is, what is the role of the deltoid in the shoulder stability? First of all, ex vivo experiments are necessary to analyse its effect. The aims of this study were: (1) to propose a testing protocol to measure the glenohumeral joint kinematics during the abduction motion by pulling on the deltoid without constraining the humerus and (2) to evaluate the repeatability of the 3D measurements. Six fresh-frozen anatomic specimens were tested. The kinematics follow-up of the osseous parts was carried out using an optoelectronic system (Polaris((R)), NDI, Canada). The abduction motion is realized by the pulling on anterior and medium fibers of the deltoid. For a 25 mm displacement, the range of motion: for the abduction was 24 degrees to 30.5 degrees , for the flexion was 1.5 degrees to -30.5 degrees (extension), for the medio-lateral rotation was 12 degrees (lateral rotation) to -5 degrees (medial rotation). For a displacement of the whole acromion-clavicle between 0 and 25 mm, the three humeral head translations were less than 5 mm. The three rotations and three translations were (with SD 95%): abduction: 0.5 degrees , flexion: 1 degrees , medio-lateral rotation: 1.5 degrees , three translations: 0.5 mm. The results showed a very high repeatability of the values. Results suggest that the deltoid alone can realize a motion of lateral elevation with a good stability in the glenohumeral joint as shown by the slight translation motion of the head and the value reproducibility. The protocol can be used to validate a finite element model of the glenohumeral joint. PMID- 17460814 TI - Real time PCR for detection of Chlamydophila pneumoniae in peripheral blood mononuclear cells of patients with multiple sclerosis. PMID- 17460815 TI - Primary malignant lymphoma of the cranial vault. AB - A 60-year-old woman presented with a subcutaneous mass on her scalp. Computed tomography (CT) showed a homogeneously enhanced mass of the parietal bone with both intra- and extra-calvarial extension and having destroyed the right parietal bone. The mass was hypointense on the T1-weighted magnetic resonance image, slightly hyperintense on the T2-weighted image and homogenously enhanced with Gd DTPA. Bone scintigraphy showed prominent accumulation of radioisotopes in the scalp lesion. The tumour was removed, including the involved bone and dura mater. Histologic diagnosis was non-Hodgkin's B-cell lymphoma, and tumour cells had infiltrated into the dura mater. The patient was treated with radiotherapy and chemotherapy. She returned to ordinary daily life and has been well without recurrence for 3 years. Although primary malignant lymphoma of the cranial vault is rare, it should be considered in the differential diagnosis when a mass is encountered in the cranial vault. We have found only fourteen such cases in the literature, and we review these cases. PMID- 17460816 TI - Predictive value of initial clinical status, intracranial pressure and transcranial Doppler pulsatility after subarachnoid haemorrhage. AB - BACKGROUND: We examined the predictive value of initial clinical status, mean arterial blood pressure (MABP), intracranial pressure (ICP) and transcranial Doppler (TCD)-derived pulsatility and resistance indices for outcome and quality of life one year following aneurysmal subarachnoid haemorrhage (SAH). METHOD: Neuromonitoring was performed in 29 patients following clipping or coiling of an aneurysm. Mean arterial blood pressure was measured in the radial artery and intracranial pressure was assessed via a closed external ventricular drainage. Based on transcranial Doppler-recordings of the middle cerebral artery, Gosling's pulsatility (PI) and Pourcelot's resistance (RI) index were calculated. Glasgow outcome score (GOS) and short form-36 (SF-36) scores were determined one year after SAH. FINDINGS: An unfavourable outcome (GOS 1-3) was observed in 34% of patients and correlated significantly (p < 0.05) with a poor initial clinical status, as determined by Glasgow Coma Scale (r = 0.55), Hunt and Hess (r = 0.62), World Federation of Neurosurgical Societies (WFNS) (r = -0.48) and Fisher (r = -0.58) score. Poor outcome was significantly associated with high mean arterial blood pressure (r = -0.44) and intracranial pressure (r = -0.48) as well as increased pulsatility (r = -0.46) and resistance (r = -0.43) indices. Hunt and Hess grade > or = 4 (OR 12.4, 5-95% CI: 1.9-82.3), mean arterial blood pressure > 95 mmHg (19.5, 2.9-132.3), Gosling's pulsatility >0.8 (6.5, 1.6-27.1) and Pourcelot's resistance >0.57 (15.4, 2.3-103.4) were predictive for unfavourable outcome in logistic regression, however TCD-diagnosed vasospasm was not. Except for mental health, significantly reduced scores were observed in all short form 36 domains. Initial clinical status correlated significantly with the physical functioning, role physical, bodily pain, social functioning and physical component summary of short form-36. CONCLUSIONS: Mortality and morbidity following SAH remains high, especially in poor-grade patients. Outcome is mainly correlated with initial clinical status, mean arterial blood pressure, intracranial pressure, pulsatility and resistance indices. Those factors seem to be stronger than the influence of vasospasm. PMID- 17460817 TI - Multiple Aspergillus brain abscesses in immuno-competent patient with severe cranio-facial trauma. AB - Aspergillosis of the central nervous system (CNS) is a rare, but well described disease in immuno-competent patients. We present a 65-year-old patient who developed neuro-aspergillosis 10 months after severe cranio-facial trauma (Le Fort III). He was treated successfully with surgery including stereotactic drainage and, with Amphotericin B, Liposomal Amphotericin B, and Itraconazol. PMID- 17460819 TI - Asylum application denied for HIV-positive Venezuelan. PMID- 17460818 TI - VII-VIII nerve complex hung up by the subarcuate artery: a cause of hemifacial spasm. PMID- 17460820 TI - Prison. HIV drug denial case is referred to Mississippi courts. PMID- 17460821 TI - Sentencing. Victim's serostatus disallowed as defense for use of force. PMID- 17460822 TI - Kenyan denied asylum based on his HIV-positive status. PMID- 17460823 TI - An unusual team of cardiothoracic surgeons. Interview by Ingrid Torjesen. PMID- 17460824 TI - History of cardiology: Sir John Parkinson, MD. PMID- 17460826 TI - Cryo-therapy in granuloma pyogenicum. PMID- 17460825 TI - The "sign of groove", a new cutaneous sign of internal malignancy. AB - A 36-year-old young male with multiple heterosexual contacts presented with bilateral inguinal bubo and the classical "sign of groove". A diagnosis of lymphogranuloma venereum (LGV) was made and a three-week course of doxycycline was given. Lack of response prompted us to investigate further. A biopsy of the bubo was consistent with non-Hodgkin's lymphoma (NHL). Immunohistochemistry of the lymph node done at the Regional Cancer Center (RCC), Trivandrum, confirmed the diagnosis as NHL of diffuse large B-cell type. The second patient, a 32-year old male with two unprotected heterosexual contacts presented with a left-sided inguinal bubo of six weeks duration. An empirical course of doxycycline was given even though investigations did not reveal any STI. Lack of response prompted us to do a lymph node biopsy, which was consistent with NHL, which later with immunohistochemistry was confirmed as NHL, diffuse large cell type. We are reporting here that the "sign of groove" is not specific for LGV as thought earlier, but can occur in NHL also. PMID- 17460827 TI - Genes versus cancer. PMID- 17460828 TI - Tough sell. PMID- 17460829 TI - Simvastatin + ezetimibe: a combination with no proven advantages. PMID- 17460830 TI - The concentration of anthranilic acid in saliva of orthodontic appliances. AB - PURPOSE: Anthranilic acid is an important, the aromatic intermediate in the degradation of tryptophan in kynurenine pathway. This compound plays an important role in the regulation of immunological processes as well shows antibacterial activity. The aim of our study was to estimate the concentration of anthranilic acid in saliva of young patients with orthodontic apparatus. We also assessed correlation between saliva anthranilic acid concentrations and time of orthodontic treatment. For the first time we have demonstrated the enhanced concentration of anthranilic acid in saliva of young orthodontic appliances. MATERIAL AND METHODS: The study was performed on non-stimulated, mixed saliva of patients with orthodontic appliances. The concentration of anthranilic acid and was determined by high-performance liquid chromatography. RESULTS: The concentration of anthranilic acid was significantly higher in orthodontic patients (p = 0.043) in comparison to healthy volunteers. The mean time of orthodontic treatment was 15.0 +/- 2.03 months. We did not observe existence of correlation between anthranilic acid concentration in saliva and time of orthodontic treatment (r = -0.250; p = 0.517). CONCLUSION: These results might indicate that anthranilic acid can be one of many factors initiating of periodontal disease in orthodontic appliances. PMID- 17460831 TI - The saliva immunology mechanisms and periodontal status in HIV infected subjects. AB - PURPOSE: The aim of this study was the evaluation of connection between parodontium determined by using GI and PBI indexes and specific immunity status and non-specific in HIV infected group and in control group. MATERIAL AND METHODS: The study was carried out in the group of 37 patients infected with HIV. Mixed non-stimulated saliva was used for the study. Peroxidase activity was determined using the method by Mansson-Rahemtull. Lysozyme and A, G, M antibodies concentrations were determined with the use of radial immunodiffusion method. The concentration of lactoferrin was determined by using ELISA method. The clinical state of parodontium estimated by means of GI and PBI evaluating quality changes in the gum. RESULTS: Deterioration of the immunological status of subjects was accompanied by the increase of the values of GI and PBI. The strong negative correlation between GI and PBI and the concentration of lactoferrin and positive activity of the peroxidase in the whole examined population was determined. In the infected group the correlation between the status of gingiva expressed by GI and concentration or activity of examined enzymes and immunoglobulins was not ascertained. CONCLUSIONS: 1. HIV infection is connected to worsening of paradontium status expressed by values of GI and PBI indexes. 2. Paradontium status correlated positively with immunological status of HIV positive subjects. 3. In HIV infected group, no connection between number of IgA, IgG, IgM, concentration of lysozyme, lactoferrin, activity of peroxidase and paradontium status was observed. PMID- 17460832 TI - Transient oral cavity and skin complications after mucositis preventing therapy (palifermin) in a patient after allogeneic PBSCT. Case history. AB - PURPOSE: The aim of this study was to assess the state of oral mucosa in a patient after allo-PBSCT who has received palifermin, a recombinant human keratinocyte growth factor. MATERIAL AND METHODS: A 19-year-old male was treated in the Department of Haematology of the Medical University in Warsaw due to the AML. Conditional chemotherapy was applied, according to the BuCy 4 + ATG regimen and allogeneic haematopoietic cells transplantation from an unrelated donor. He was receiving palifermin intravenously for 3 consecutive days immediately before the initiation of conditioning therapy and after allogeneic PBSCT. On day +3 the oral mucous membrane was pale and swollen, with linea alba visible on cheeks. Superficial glossitis and viral pharyngitis were noted. Beginning with day +5/+6 proliferative gingivitis was observed. On day +9 gingival contour was altered and the gingiva covered nearly completely tooth crowns of all teeth. The gingiva were whitened, as if covered by thick epithelium. Slight gingival hyperplasia was still observed on day +24. Since day +4/+5 skin rash coexisted, spreading over hairy head skin, face, dorsum and chest. Disseminated papulopustular (acne-like) lesions were observed, some of them related to the hair follicles. Skin changes were present till day +15. CONCLUSIONS: Palifermin is an efficient pharmaceutical in mucositis prevention in patients after allogeneic PBSC transplantation. Transient complication of hyperplastic gingivitis with a concomitant skin eczema of a papulopustular nature arose. PMID- 17460833 TI - Periodontal condition in patients with cardiovascular diseases. AB - The cardiovascular system diseases constitute a serious problem for modern medicine. THE AIM: To investigate the potential risk and the connection of periodontal diseases and cardiovascular disorders. MATERIAL: The examination was performed in the group of 104 patients of both sexes, aged 50-90 years. The patients were divided into two groups: group I--patients with hypertension (47 subjects), group II--patients with fresh myocardial infarction, treated with primary coronary angioplasty (57 subjects). METHODS: The OHI index, according to Greene and Vermillion, was used to assess the oral hygiene and periodontal clinical conditions were evaluated according to Russell's PI index, modified by Davies. CPI index was used to estimate the state of periodontium. Teeth loss was classified according to the Eichner's classification. RESULTS: The value of OHI index differs in both groups. Highest value was registered at 5 patients in the I group vs 2 in the II group. Lowest value was recorded in 11 patients in the I group and 4 in the II group. The value 0.0-0.2 PI was recorded at 14 persons in the I group and 11 in the II group. The value 1.6-3.8 of PI index was registered at 2 in the I group and 6 in the II group. Healthy periodontium was stated in 10 patients with hypertension and only 2 with myocardial infarction. The CPI = 2 was shown in 12 patients with hypertension and 11 with myocardial infarction, CPI = 3 was shown in 23 patients with myocardial infarction. CONCLUSION: The studies revealed bad condition of the oral cavities of patients with hypertension, and specifically with fresh myocardial infarction. PMID- 17460835 TI - The evaluation of CPITN index among adults living in Podlasie region. AB - The aim of this study was to evaluate the condition and treatment needs of the periodontium in adults living in Podlasie region. Checked population was divided into three groups: 18 year old, 35-44 and 65-74 year old. The assessment of the periodontium status was performed on the basis of CPITN index. The study showed that young people usually did not need any periodontal treatment. The predominating treatment need was removing of dental calculus, respectively 7.4% subjects aged 18, 62.5% of second group and 58.7% of the oldest one. 10% persons aged 35-44 and 6.9% persons aged 65-74 required complex periodontal treatment. The number of excluded sextants grown with aged. PMID- 17460834 TI - Short time effect of elmex and Listerine mouthrinses on plaque in 12-year-old children. AB - PURPOSE: This study was conducted to determine the effect of two mouthrinses elmex and Listerine on plaque accumulation in 12-year-olds. MATERIAL AND METHODS: 30 12-year-old children took part in the clinical study. They were divided into three groups. Group I (10 people) was given Listerine to home use. Group II (10 people) was given elmex to home use. Group III (10 people) did not receive any mouthrinses. Following indices were used in first and base study Plaque Index (PI), Approximal Plaque Index (API) and Sulcus Bleeding Index (SBI). The statistical analysis was performed using T test for related samples and Spearman rank order correlations. RESULTS: Mean PI lessened in group I (Listerine) from 0.996 to 0.804 and group II (elmex) from 0.807 to 0.698. In group III it stayed almost at the same level. In all children values of API and SBI decreased after two weeks. Reduce of API in participants using Listerine was important statistically and it lessened from 57.4% to 48.1% (reduction by 16.2%). The other results of API and SBI were not statistically important. API in children using elmex lowered by 15.5%. Bleeding (SBI) in Listerine group decreased by 21.5% and in elmex group decreased by 24.5%. In control group diminish of SBI was only by 14.4%. CONCLUSIONS: In summary, this study has demonstrated that additional rinsing helped in reducing plaque and gingivitis in 12-year-olds but it is not as essential as motivation to everyday oral hygiene. PMID- 17460836 TI - Hygienic habits and the dental condition in 12-year-old children. AB - PURPOSE: The aim of the study was the description of the dental condition of hygienic routines in 12-year-old children in urban and rural areas of Lublin voivodship. MATERIAL AND METHODS: The study comprised 274 children at the age of 12 (152 girls and 122 boys). 95 girls and 92 boys came from the urban area; 57 girls and 30 boys came from the rural area. RESULTS: On the basis of clinical examination it was concluded that 11.96% of boys and 18.95% of girls from the urban area and 6.67% boys and 8.77% of girls from the rural area brush their teeth after every meal; 60.87% of boys and 68.42% of girls from the urban area and 43.33% of boys and 50.88% of girls from the rural area brush their teeth twice; 22.83% of boys and 11.58% of girls from the urban area and 26.67% of boys and 28.07% of girls from the rural area brush their teeth once daily. DMF count was for boys from the urban area--4.12/girls--3.92 and for boys from the rural area 4.50/girls--4.29. The treatment indicator was for boys from the urban area- 0.56/ girls - 0.47 and for boys from the rural area 0.35/girls--0.67. CONCLUSIONS: On the basis of the research conducted in the study, it was concluded that tooth brushing is more frequent with urban area children than in children from rural area. This leads to a conclusion that the action for improvement of the health awareness is a dire need among 12-year-olds both from urban and from rural areas. PMID- 17460837 TI - Prevalence rate and antibiotic susceptibility of oral viridans group streptococci (VGS) in healthy children population. AB - PURPOSE: The aim of this study was to evaluated the prevalence rate of oral viridans group streptococci (VGS) and their susceptibilities to some antibiotics in healthy children. MATERIAL AND METHODS: Samples of pharyngeal swabs and supragingival dental plaques for microbiological studies were collected from 206 healthy children, aged 4-18 years. Additionally, 75 samples of carious lesions from children with dental caries were included. The streptococci were isolated and identified using standard methods and commercial identification kits. For performance of antibacterial susceptibility testing of VGS strains disk diffusion and/or breakpoints procedures were used according to NCCLS standards and criteria. A total of 425 VGS strains were tested against penicillin, ampicillin, erythromycin, clindamycin, tetracycline, doxycycline, ciprofloxacin and vancomycin. RESULTS: A total of 239 VGS strains belonging to 8 species from pharyngeal swabs of 192 (93.2%) children were isolated. VGS strains from supragingival plaques were isolated in 149 (72.3%) healthy children (p < 0.05), and from carious lesions in 37 (49.3%) children with dental caries. VGS strains of S. mitis species were isolated most frequently from 4-5 year old as compared to 12 and 18 year old children (p < 0.05), while S. vestibularis strains isolated most often in 12 year old ones (p < 0.05). Among 425 VGS strains, high level of penicillin resistance (MIC > or = 2.0 mg/L) was shown in 71 (16.7%) strains, 33 (46.5%) of them belonged to S. mitis species. VGS strains were also resistant to erythromycin (23.5%), clindamycin (23.1%), tetracyclines (T-52%, DOX-16%), gentamycin (25.9%) and ciprofloxacin (55.2%). All VGS strains were vancomycin - susceptible. CONCLUSIONS: 1. In the oral cavities of healthy children, approximately 98% of streptococci belonged to two VGS groups, i.e. mitis and salivarius groups. Streptococci of mutans and anginosus groups were isolated sporadically (2%). 2. We observed difference in susceptibility to penicillin and other antibiotics between the various species of viridans groups streptococci. Mitis group strains (except S. pneumoniae) were more frequently penicillin resistant (23%) in comparison to salivarius group of VGS strains (9%) (p = 0.0001). PMID- 17460838 TI - Assessment of the state of dentition and oral hygiene in 16-25-year-old young people with mild and moderate mental disability. AB - PURPOSE: The purpose of the research is to assess the state of dentition and oral hygiene in 16-25-year-old young people with mild and moderate mental disability in comparison with a control group of healthy young people at the same age. MATERIAL AND METHODS: The research was carried out in a special School and Tutelary Centre in Lublin. A group of 144 young people aged 16-25 with mild and moderate mental disability (group I) among them 75 girls and 69 boys participated in the research. A group of 50 healthy young people aged 16-25 (group II) among them 24 girls and 26 boys was a control group. Determined: frequency of dental caries, DMF number, dental caries treatment index (DTI), oral hygiene index (OHI), percentage of traumatic injuries of teeth, percentage of sealed teeth. RESULTS: The frequency of dental caries in both groups was 100%. The average DMF was 11.96 (group I) and in the control group II: 6.58. The largest number of teeth with active caries--8.21 teeth with caries per person was found in group I, but 2.72 in group II. Dental caries treatment index (DTI) was 0.24 in group I and 0.59 in the control group II. Oral hygiene index OHI in group I was 1.78, in group II this index was 0.34, 0.29 in girls and 0.38 in boys. CONCLUSIONS: 1. The state of dentition in 16-25-year-old young people with mild and moderate mental disability is unsatisfactory. 2. Higher values of OHI index were in group I. 3. The obtained results of the state of dentition and oral hygiene in the group of young people with mental disability are at the same level both in the girls and boys. 4. The above mentioned results suggest the need for special dental care for young people with mild and moderate mental disability. PMID- 17460839 TI - Microorganisms in root carious lesions in adults. AB - PURPOSE: Root caries is emerging as a significant problem in the middle aged and elderly subjects because of the improving general health conditions, and medical and technological advances. The purpose of this investigation was to assess the prevalence of aerobic and anaerobic bacteria as well as yeasts of Candida genus in root carious lesions in middle-aged and older adults. MATERIAL AND METHODS: Specimens of root carious lesions were collected from 78 adults for bacteriological and mycological studies. Standard procedures of culture, isolation, and identification of aerobic and anaerobic bacteria, and fungi were used in the study. RESULTS: The analysis of results was performed independently in two age groups of adults, i.e. 52 subjects aged 35-44 years (middle age) and 26-aged 55-72 years (older age). There were 120 bacterial strains isolated from root carious lesions in middle-aged subjects, 63 (52.5%) strains belonged to 5 genera of aerobic bacteria and 57 (47.5%)--to 7 genera of anaerobic bacteria (p > 0.05). While in the second group, 85 strains were isolated, 54 (63.5%)--6 genera of aerobic bacteria and 31 (36.5%)--4 genera of anaerobic bacteria (p = 0.0004). There were no differences between the isolation rate of a various species in both examined groups, except for Streptococcus spp., S. oralis, Micrococcus spp., Neisseria spp. and Veillonella spp., which statistically significantly most frequent occurred in elderly (p < 0.05). The yeasts of Candida genus of 4 species (C. albicans, C. lusitaniae, C. pelliculosa, and C. pulcherrima) were isolated from middle-aged subjects (32.7%) with the comparable rate to older adults (30.8%; only C. albicans) (p > 0.05). Among all isolated microorganisms, Candida spp., were comprised about 10% in both examined groups (p > 0.05). CONCLUSIONS: Aerobic Gram-positive cocci (Staphylococcus spp. and Streptococcus spp.) as well as anaerobic ones (Peptostreptococcus spp.), and Candida albicans were occurred most frequently in root carious lesions in middle-aged and older adults. PMID- 17460840 TI - Fundraising initiatives and vendor solicitation: avoiding kickback implications. PMID- 17460841 TI - Welcome to Medicare visit and AAA screening. PMID- 17460842 TI - Semiformal attire and a smile. PMID- 17460843 TI - Regarding "efficacy of stellate ganglion blockade for the management of type 1 complex regional pain syndrome". PMID- 17460844 TI - The body as a battlefield: stories from the front line of spiritual health. PMID- 17460845 TI - Evaluation of penicillin allergy without benzylpenicilloyl-polylysine. PMID- 17460846 TI - The strategic importance of thinking time: creating a mind-set for safety. PMID- 17460848 TI - Direct reporting by patients: positive results. PMID- 17460847 TI - Faculty Matters. Gary L Loving. PMID- 17460849 TI - It's all about you: my resolutions for 2007. PMID- 17460850 TI - Publishing--part II. PMID- 17460851 TI - Deferiprone: agranulocytosis and neurological disorders. PMID- 17460852 TI - Commentary. Brief communication: ramipril markedly improves walking ability in patients with peripheral arterial disease: a randomized trial. PMID- 17460853 TI - Commentary. Emergency endovascular stent-graft treatment for acute thoracic aortic syndromes. PMID- 17460854 TI - Commentary. Advancements in carotid stenting leading to reductions in perioperative morbidity among patients 80 years and older. PMID- 17460855 TI - Commentary. Is carotid artery stenting in octogenarians really dangerous? PMID- 17460857 TI - Bibliography. Current world literature. Vascular biology. PMID- 17460856 TI - Commentary. Synthetic vascular prosthesis impregnated with mesenchymal stem cells overexpressing endothelial nitric oxide synthase. PMID- 17460858 TI - Commentary. Clonidine decreases stress response in patients undergoing carotid endarterectomy under regional anesthesia: a prospective, randomized, double blinded, placebo-controlled study. PMID- 17460859 TI - Arterial 'prehypertension': a useful concept for drug companies, useless for patients. AB - (1) Elevated blood pressure is an independent and progressive cardiovascular risk factor. The risk starts to increase above a threshold of 115/75 mmHg. (2) The threshold values for blood pressure with practical implications for patients' health have been determined from clinical trial results. These are, for example, 160/95 mmHg in patients without diabetes and complications of hypertension, and 140/80 mmHg in patients with diabetes or a history of stroke. (3) A prospective cohort analysis confirmed the progressive nature of the relation between blood pressure and the risk of cardiovascular events: after about 12 years the incidence of cardiovascular events was 7% when blood pressure was less than 120/80 mmHg and 12% when it was between 130/85 and 140/90 mmHg. However, patients with these moderately elevated blood pressure values were also more likely to be diabetic. (4) The only trial involving patients with systolic pressure values between 130 and 139 mmHg, levels referred to by some as 'prehypertension', was not designed to determine either the clinical benefits or the adverse effects of treatment with candesartan. Two years after withdrawal of this antihypertensive drug, there was no statistically significant difference in the proportion of patients requiring antihypertensive treatment (threshold 160/100 mmHg). (5) In practice, 'prehypertension' is not a useful concept for patient management. The blood pressure thresholds above which the risk-benefit balance for some treatments becomes positive, in terms of morbidity or mortality, remain at 160/95 mmHg for patients without diabetes or complications and 140/80 mmHg for patients with diabetes or a history of stroke. PMID- 17460860 TI - Chickenpox: very low mortality in France and the USA. PMID- 17460861 TI - Re: "Work related stress and pain". PMID- 17460862 TI - Enter the health zone: how summer camps introduce students to health care careers. PMID- 17460863 TI - Tail docking regulations go through. PMID- 17460866 TI - The truth about 'nano'. Interview by Nicola Nugent. PMID- 17460864 TI - Concern about the sudden withdrawal of brucellosis testing. PMID- 17460867 TI - DNA block copolymers. PMID- 17460868 TI - Maternal mortality and related concepts. AB - OBJECTIVE: This report presents data on U.S. deaths to pregnant or recently pregnant women, summarizes long-term processing issues, and examines recent changes affecting the data and the impact of the changes on the statistics for these women. METHODS: This report presents descriptive tabulations of information reported on death certificates that are completed by funeral directors, attending physicians, medical examiners, and coroners. The original records are filed in the state registration offices. Statistical information is compiled into a national database through the Vital Statistics Cooperative Program of the Centers for Disease Control and Prevention's (CDC) National Center for Health Statistics (NCHS). Causes of death are processed in accordance with the International Classification of Diseases (ICD). RESULTS: Maternal mortality fluctuates from year to year but was 12.1 deaths per 100,000 live births in 2003. The implementation of the International Classification of Diseases, Tenth Revision (ICD-10) in 1999 resulted in about a 13 percent increase in the number of deaths identified as maternal deaths between 1998 and 1999. The rate increased again between 2002 and 2003 after a separate pregnancy question became a standard item on the U.S. Standard Certificate of Death. The adoption of a standard separate question on pregnancy facilitates the identification of late maternal deaths. CONCLUSION: Maternal deaths increased with the introduction of the ICD-10 and with changes associated with the addition of a separate pregnancy status question on the U.S. Standard Certificate of Death. These changes may result in better identification of maternal deaths. PMID- 17460869 TI - Health care technology assessment: implications for modern medical practice. Part I. Understanding technology adoption and analyses. AB - In the modern era of rapidly rising medical costs, health care technology assessment--multidisciplinary evaluation of clinical and economic aspects of technology--has assumed an increasingly important role in health policy and clinical decision-making. This review examines health care technology adoption, its impact on medical and surgical practice, and recent trends in health care technology assessment. Part I discusses the difficult challenges posed by assessment and provides a guide to the methodologies used. PMID- 17460870 TI - The orthopedist as clinical densitometrist: cost- and time-effectiveness. AB - We tested the hypothesis that an orthopedic surgeon and his or her staff can efficiently and economically provide a bone densitometry service. This hypothesis reflects a philosophy that orthopedists should take a more active role in identifying patients at risk for osteoporosis. We evaluated the cost- and time effectiveness of an orthopedic surgeon and his medical assistant in completing reports and related correspondence for dual-energy x-ray absorptiometry scans conducted in an orthopedic subspecialty clinic. Cost analysis showed that completing 14 or 15 reports per month was required to break even and that completing up to 40 reports per month was a highly efficient and economic use of the surgeon's time. PMID- 17460871 TI - Flexible intramedullary nailing for a segmental radial fracture of the neck and shaft in a child. PMID- 17460872 TI - What residents need. PMID- 17460873 TI - Blastomyces dermatitidis osteomyelitis of the tibia. PMID- 17460874 TI - Distal biceps tendon repair: anchor versus transosseous suture fixation. AB - Suture anchor fixation and transosseous suture fixation were compared in 12 fresh frozen cadaveric radii using either No. 2 braided polyester suture or single Mainstay 3.5-mm threaded anchors (made at the time by Howmedica, Rutherford, NJ) with No. 2 suture. Suture fixation failed at a mean strength of 162 N (range, 129 179 N), anchor fixation at 136 N (range, 121-150 N). Neither technique is strong enough to safely allow immediate biceps activity. Nevertheless, suture anchor fixation to the radial tuberosity offers a lower but clinically comparable strength to transosseous suture fixation while limiting postoperative risks. PMID- 17460875 TI - Assessment of acetabular version by plain radiograph. AB - Radiographs are routinely used to assess the condition and position of the acetabular component. The condition of the cement mantle, or the ingrowth potential, is usually easily recognized. Component-bone position can be assessed by using the method of Ranawat or by measuring abduction angles. Assessment of the version of an acetabular component is often overlooked. This angle or position is important relative to instability, impingement, and motion abnormality. The opening angle or version can be implied from a true acetabular or cross-table lateral radiograph, but good-quality views are often difficult to obtain on an outpatient basis. Using the simple technique presented here, clinicians can assess the acetabular component for version on the basis of plain anteroposterior pelvis and hip radiographs. PMID- 17460876 TI - Early prediction of myocardial salvage after primary coronary angioplasty: comparative study of coronary flow velocity pattern immediately after primary coronary angioplasty and perfusion-metabolism mismatch. AB - OBJECTIVES: Perfusion-metabolism mismatch in the subacute phase using thallium 201/radio iodinated beta-methyl-p-iodophenyl pentadecanoic acid (T1/BMIPP) dual scintigraphy is an indicator of viable myocardium in acute myocardial infarction. This study investigated early prediction of myocardial salvage from the T1/BMIPP mismatch and coronary flow velocity (CFV) patterns in patients with acute myocardial infarction. METHODS: Thirty three patients with first anterior wall myocardial infarction underwent primary coronary angioplasty and achieved reflow within 8 hr of onset. By using a Doppler guide wire, CFV patterns were assessed immediately after primary coronary angioplasty. T1/BMIPP dual scintigraphy was performed within 3 days after reperfusion. The extent of discordance in severity score was defined as the T1/BMIPP mismatch score. RESULTS: Regression analysis showed dual scintigraphy mismatch score correlated well with deceleration time of diastolic flow velocity (r = 0.54, p < 0.01). Mismatch score was greater in the non-early systolic reversal flow group than in the early systolic reversal flow group (5.5 +/- 3.3 vs 1.9 +/- 2.1, respectively, p < 0.01). CONCLUSIONS: Changes in CFV patterns correlated well with T1/BMIPP mismatch score. CFV pattern immediately after reperfusion is useful for early prediction of myocardial salvage. PMID- 17460877 TI - Possibility of close relationship between sleep disorder breathing and acute coronary syndrome. AB - OBJECTIVES: Sleep apnea syndrome and acute coronary syndrome (ACS) are related, but any further association with congestive heart failure (CHF) remains unclear. METHODS: Sixty-five patients with ACS (ACS group) and 48 patients with CHF (CHF group)underwent Holter electrocardiography and respiratory monitoring to identify sleep apnea. RESULTS: There were significant differences in age, sex, frequency of smoking, and ejection fraction between the two groups. The apnea hypopnea index showed similar high values in both ACS group (21.7 +/- 17.0/hr) and CHF group (19.4 +/- 17.9/hr). In the ACS group, 24 patients (37%) had central sleep apnea syndrome and 29 patients (45%) had obstructive sleep apnea syndrome. There were no significant differences in the incidences of central and obstructive sleep apnea syndromes between the two groups. Sympathetic nerve activity was significantly higher in ACS group than in CHF group (low/high frequency power ratio in overall study, 2.64 +/- 2.43 vs 1.24 +/- 1.05, p = 0.0003; in asleep study, 2.64 +/- 2.35 vs 1.23 +/- 1.04, p = 0.0002; in awake study, 2.73 +/- 2.36 vs 1.50 +/- 1.46, p = 0.002). CONCLUSIONS: Sleep apnea was observed at the same frequency in the ACS group and the CHF group including higher sympathetic nerve activity, and there was no significant difference in frequency of desaturation. This study suggested that sleep disorder breathing is frequently and similarly associated with both CHF and ACS. PMID- 17460878 TI - Defect images in stress thallium-201 myocardial scintigraphy in patients with complete left bundle branch block: comparison of exercise stress and pharmacological stress. AB - OBJECTIVES: Stress thallium-201 (201Tl) myocardial scintigraphy can demonstrate perfusion abnormalities, especially in the septum in patients with complete left bundle branch block (CLBBB) even with angiographically normal coronary arteries. Differences in the images between exercise and pharmacological stress 201Tl myocardial scintigraphy were evaluated in patients with CLBBB and normal coronary arteries. METHODS: Forty-five patients with CLBBB underwent exercise stress using treadmill or pharmacological (adenosine triphosphate) stress 201Tl myocardial scintigraphy from October 1997 to February 2003. Patients with myocardial diseases were excluded, such as cardiomyopathy and coronary artery diseases detected by echocardiography and/or cardiac catheterization. The myocardial segment was classified according to the American Heart Association style for coronary artery disease. RESULTS: Peak blood pressure levels and heart rates were significantly higher in the exercise stress group than in the pharmacological stress group (p < 0.001). The rate of defects in stress images was significantly higher in the exercise stress group (72.4%; 21/29 cases) than in the pharmacological stress group (18.8%; 3/16 cases) (p < 0.01). The rate of redistribution of observed defects in delayed images was 76.2% (16/21 cases) in the exercise stress group, and 0% (0/3 cases)in the pharmacological stress group (p < 0.01). The myocardial segments showing defects were different between the exercise stress group and the pharmacological stress group. CONCLUSIONS: Patients with CLBBB showed different frequencies of defects by stress 201Tl myocardial scintigraphy according to the stress method. Moreover, defects also occured in areas other than the septum. Blood pressure and heart rate were involved in the mechanisms of defects in left bundle branch block. PMID- 17460879 TI - Thermal therapy improves left ventricular diastolic function in patients with congestive heart failure: a tissue doppler echocardiographic study. AB - OBJECTIVES: We previously reported that systemic thermal therapy using 60 degrees C dry sauna improves left ventricular systolic function and clinical symptoms in patients with chronic heart failure. The aim of this study was to investigate the effects of thermal therapy on left ventricular diastolic function. METHODS: We examined transmitral inflow and mitral annular velocity before and after sauna in 10 patients with congestive heart failure using pulsed and tissue Doppler echocardiography. RESULTS: Left ventricular and left atrial dimensions and left ventricular percentage fractional shortening did not change after sauna. Early diastolic mitral inflow velocity (E) increased and the deceleration time of the E wave decreased significantly after sauna compared to before sauna. Early diastolic mitral annular velocity (E') significantly increased after sauna. The deceleration time of E' significantly decreased after sauna compared to before sauna. The E/E' significantly decreased 30 min after sauna. CONCLUSIONS: Thermal therapy improves acute left ventricular diastolic function in patients with congestive heart failure. PMID- 17460880 TI - Ultrasound attenuated coronary plaque as a risk factor for slow flow or no-reflow during percutaneous coronary intervention: a case report. AB - Slow flow or no-reflow is a serious complication during percutaneous coronary intervention (PCI), but little is known about the risk factors. A 64-year-old man underwent coronary angiography and PCI for stable angina. Pre-interventional intravascular ultrasound demonstrated an ultrasound attenuated coronary plaque, as a long eccentric bulky plaque with a marked decrease of the back echo without calcification. Since the lesion was highly eccentric in the large left anterior descending artery, directional coronary atherectomy (DCA) and subsequent stent implantation were planned. Serious no-reflow occurred after DCA. The DCA specimen suggested that the lipid-laden atheromatous gruel could attenuate the ultrasound reflection and cause distal embolization, resulting in no-reflow during PCI. The presence of ultrasound attenuated coronary plaque is a predictor of slow flow or no-reflow in PCI, indicating that distal protection devices may be required during the procedure. PMID- 17460881 TI - [Papillary fibroelastoma in the right ventricular outflow tract: a case report]. AB - A 60-year-old female was admitted to our hospital for further examination of heart murmur. Chest radiography revealed cardiomegaly and pulmonary congestion. Two-dimensional echocardiography showed a mobile and pedunculated mass, approximately 20 mm in diameter, attached to the right ventricular outflow tract, and perimembranous ventricular septal defect. Cardiac catheterization and blood sampling were performed, which showed a pulmonary to systemic flow ratio of 3.70. She underwent surgical excision of the tumor, ventricular septal defect patch suture and tricuspid annuloplasty. Histological examination confirmed papillary fibroelastoma. The predominant location of papillary fibroelastoma is the valvular surface. This very rare case of papillary fibroelastoma was located in the right ventricular outflow tract. PMID- 17460882 TI - [Primary effusion lymphoma complicating cardiac tamponade: a case report]. AB - A 76-year-old woman was admitted to our hospital because of exertional dyspnea and leg edema during the previous month. Her systolic blood pressure on admission was 80 mmHg with 12 mmHg of pulsus paradoxous, and her pulse rate was 110 beats/min. Chest radiography revealed marked cardiomegaly and echocardiography showed massive pericardial effusion mainly behind the left ventricle and collapse of the right ventricle. The initial diagnosis was pericardial tamponade. Pericardiocentesis and pericardial drainage revealed bloody pericardial effusion. After drainage, her vital signs improved and her symptoms immediately disappeared. The cytological analysis of the pericardial effusion revealed numerous lymphoma cells. Computed tomography of the neck, chest and abdomen showed no evidence of tumor masses, lymph node enlargement, or hepatosplenomegaly. Infectious disease, collagen disease and aortic dissection were excluded. The final diagnosis was primary effusion lymphoma. The prognosis of primary effusion lymphoma is generally unfavorable because it is frequently accompanied by immunodeficiency disease. However, there was no human immunodeficiency virus infection in this patient. Fortunately, the effect of chemotherapy was excellent and the patient is doing well 1 year after the diagnosis. PMID- 17460883 TI - [Epigastric pain with elevated fibrin degradation product-DD]. PMID- 17460884 TI - Clinical application of real-time three-dimensional echocardiography. PMID- 17460885 TI - [The product specificity evolution of cyclodextrin glucanotransferase: problems and challenges]. AB - Cyclodextrin glucanotransferase, the essential enzyme for the production of cyclodextrins, has become the focus of scientific research nowadays. Although many related enzyme properties are well known, the crucial factors in product specificity determination remain to be answered. Here, the recent research progresses of cyclodextrin glucanotransferase, especially those about the evolution of product specificity, were reviewed, and the scientific problems were discussed. PMID- 17460886 TI - [Structure-based design and biosynthesis of collagen proteins]. AB - Collagen is the most abundant protein in human body and a periodic helix, i. e. , triple helix, fibrous protein, which provides the scaffold structures for the cell adhesion and macromolecule aggregation, etc. With the development of gene engineering and biomaterial technologies, and the incessant studies on the technique to obtain the proteins with special functions, the collagen protein has been one of the third generation biomaterials that attract more attention than others. In this paper, we reviewed the recent structure-based design and biosynthesis of collagen. PMID- 17460887 TI - [Multipotential differentiation and potential applications of adipose-derived stem cells]. AB - Adipose tissue contains a population of multipotent cells called adipose-derived stem cells (ADSCs). With the similar properties of marrow-derived mesenchymal stem cells, ADSCs have the ability to differentiate differentiate towards adipogenic, osteogenic, chondrogenic, myogenic, endothelial, hematopoietic, hepatic, islet, and neurogenic cell lineages. As adipose tissue in harvested in large amounts with minimal morbidity, it can be widely used in tissue engineering, organ repair and gene therapy. This paper focused on the plasticity of ADSCs and reviewed the new advances of this field. Finally, the problems and prospect for application was also discussed. PMID- 17460888 TI - [Progress in metabolism and crucial enzymes of glycerol conversion to 1,3 propanediol]. AB - 1,3-propanediol production by microbial fermentation has become the research hot spot for its amiability with the environment. Here the molecular mechanism of glycerol bioconversion to 1,3-propanediol was outlined by elucidating the fermentation strains, metabolic pathways, regulon and key enzymes. Of enzymes, glycerol dehydrogenase, the velocity-limiting enzyme in glycerol reductive pathway, was emphatically discussed with regard to its molecular structure and reactivating factors. This paper aims to provide the basis for genetic modification of fermentation strains. PMID- 17460889 TI - [Improved the solubility of maize uroporphyrinogen III methyltransferase as the red fluorescent indicator by site-directed mutagenesis]. AB - S-adenosylmethionine-dependent uroporphyrinogen III methyltransferase (SUMT) is a novel red fluorescence indicator. However, the production of SUMT in Escherichia coli is restricted by its relatively low solubility, and little is known about the red fluorescent materials that are associate with SUMT. Two individual SUMT mutations, L166A and L88R/L89G double mutant were produced by site-directed mutagenesis. Both mutants were overexpressed in E. coli and purified by Ni-NTA chromatography. The reddish mixtures isolated from the purified L88R/L89G double mutant were analyzed by UV-visible spectra scanning and mass analysis(MS). The L88R/L89G double mutant has enzymatic activity in vivo, whereas L166A mutant loses the activity. Trimethylpyrrocorphin is identified as the main constituent in the isolated pigments. The purified L88R/L89G mutant increases protein solubility, which is applied potentially as the fluorescent indicator denoting the solubility of protein fusion partner. PMID- 17460890 TI - [Construction and immunization of a enterohemorrhagic Escherichia coli O157 attenuated]. AB - Enterohemorrhagic Escherichia coli (EHEC) O157:H7 is an important pathogen. One of the important virulence traits of EHEC O157:H7 is the capacity to produce attaching and effacing (A/E) lesions on enterocyte. This property encoded by a pathogenicity island termed the locus of enterocyte effacement (LEE). LEE contains ler (LEE-encoded regulator) gene. The product of ler is a central up regulator of many virulence genes of the LEE. Another important virulence factor of EHEC O157: H7 is Shiga toxin (Stx), encoded by a prophage integrated into the chromosome of O157:H7. In order to obtain an attenuated vaccine candidate, a ler deletion mutant of O157: H7 was constructed by use of suicide vector pCVD442. Meanwhile, due to potential instability of the prophage carrying the stx gene, the prophage was cured with serial passages of bacteria and confirmed by PCR and DNA sequencing. A ler/stx deletion mutant of EHEC O157:H7 was constructed, termed as O157:H7(deltaler/deltastx). The cultural supernatant of O157 ler/stx deletion mutant was inoculated in vero cell culture, and the result indicating that O157 ler/stx deletion mutant lost the toxigenicity to vero cell. Test group and control group of mice were orogstrically inoculated with the O157 ler/stx deletion mutant and the virulent strain O157:H7 EDL933, respectively. Mice were observed daily for clinical signs and weight changes. After inoculation of the deletion mutant, test group of mice (inoculated with O157:H7(deltaler/deltastx)) gained weight normally and experienced no clinical signs. In contrast, control group of mice (inoculated with O157: H7) exhibited weight loss and all died in four days. In another experiment, pregnant mice were orally vaccinated by O157:H7(deltaler/ deltastx) twice at interval of 14 days. Subsequently, the suckling mice were orally challenged with O157:H7 EDL933 at 7 days of age. The results showed that 78.34% of the sucking mice born by vaccinated mice were survival and 12.73% of the sucking mice born by non-vaccinated mice were survival. This study demonstrated that O157 ler/stx deletion mutant lost the toxigenicity to vero cell and to be safety to mice. Oral immunization can induce specific immune responses, and this mutant strain could be used as an attenuated vaccine candidate against EHEC O157. PMID- 17460891 TI - [Expression and characterization of fusion protein tTF/SA as a universal effector for targeting blood coagulation]. AB - To prepare a novel fusion protein (tTF/SA) as a universal effector for targeting therapy of blood coagulation and to analyze its biological activities. The fusion gene tTF/SA was constructed by PCR, then inserted into expression vector pET22 b (+), and expressed in E. coli BL21 (DE3). The fusion protein was purified using Nickel-affinity chromatography column. The activities of tTF moiety of the fusion protein were analyzed by clotting assay and FX activation assay, and the binding activities of Streptavidin(SA) to Biotin(B) were analyzed using ELISA. RESULTS: The recombinant plasmid tTF/SA/pET22 b (+) with the correct sequence was obtained. The fusion gene tTF/SA was expressed with high yield in E. coli BL21 (DE3). The purified fusion protein retain the abilities of activating FX, inducing blood coagulation, and binding Biotin. The fusion gene tTF/SA was successfully expressed in E. coli BL21 (DE3). The recombinant tTF/SA proteins retain the activities of TF and SA. The multitarget therapy of selectively inducing thrombosis in tumor blood vessels can be achieved by the combination of tTF/SA, as a universal effector, and biotinlated carriers directing to tumor blood vessels. PMID- 17460892 TI - [The targeting study of human serum albumin gene]. AB - To generate transgenic porcine which expresses human serum albumin (HSA), the HSA gene targeting vector was constructed with HSA cDNA as the gene of interestand partial porcine serum albumin (PSA) gene as homologous arms which respectively were 7.2 kb 5' regulation sequence and 2.8 kb genomic sequence from the first intron to the fourth intron. The resistant gene neo was inserted into intron 1 and tk was ligated to the 3' end of the construct. Linearized targeting construct DNA was introduced into the fibroblast cells obtained from porcine fetus by electroporation. The positive-negative selection was performed and survival clones were screened by PCR and Southern blot. Three colonies with correct homologous recombination were obtained. Our results set a good basis for the establishment of transgenic porcine by gene target and nuclear transfer methods. PMID- 17460893 TI - [Transfection with a novel cationic gene carrier: PEI-PBLG]. AB - This study mainly deals with cell transfection and cytotoxicity for PEI(10kD) PBLG, a novel cationic copolymer, to observe its potential as a gene carrier. Size measurement and SEM were used to show the modality of the PEI-PBLG/pDNA complexes. Cytotoxicity of PEI (10kD)-PBLG was evaluated by 3-(4,5 dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) colorimetric assay and compared with PEI(25kD)-PBLG, PEI(10kD), and PEI(25kD). Furthermore, pEGFP that can express the enhanced green fluorescent protein was chosen as a reporter to observe the transfection efficiency directly. Then, PEI (10kD)-PBLG/pEGFP complexes were transfected into several cell lines, such as Hela, COS-7, Vero-E6, and ECV-304, and effects of the transfection conditions were evaluated. The efficiencies were measured by FACS. Size measurement of complex particles indicated that PEI-PBLG/pDNA tended to form smaller nanoparticles compared with PEI/pDNA. The representative size of the PEI(10kD)-PBLG/pDNA complex was approximately 100 - 200 nm. SEM images showed that the particles were condense and compact. This can be suitable for their entry into cells. Cytotoxicity studies suggested that PEI (10kD)-PBLG had considerably lower toxicity than the other three materials. In the transfection tests, PEI (10kD)-PBLG/pDNA complexes could be transfected into all the cell lines that were tested. These provided the highest level of EGFP expression (45.02%) in Hela cells, which was considerably higher than that of PEI(10kD)/pEGFP (29.16%). Being less affected by the serum during transfection, PEI-PBLG/pDNA complexes offered greater biocompatibility than PEI. PEI-PBLG copolymer reduces the cytotoxicity of PEI, improves the transfection efficiency, and offers greater biocompatibility than PEI. It shows considerable potential as an efficient nonviral carrier for gene delivery. PMID- 17460894 TI - [Construction of rat bdnf gene lentiviral vector and its expression in mesenchymal stem cells]. AB - Recently, mesenchymal stem cells (MSCs) have been one of the target cells of gene engineering. To construct the lentiviral (LV) vectors carrying the brain-derived neurotrophic factor (Bdnf) gene, the rat mesenchymal stem cells (rMSCs) were infected and finally the Bdnf gene-modified rMSCs was obtained. The CDS region of the rat Bdnf gene was obtained with reverse transcriptase-polymerase chain reaction (RT-PCR), and the transfer plasmid (PNL-BDNF-IRES2-EGFP) of the LV vector was constructed. The three plasmids of LV vector: PNL-BDNF-IRES2-EGFP, HELPER, and VSVG were cotransfected to 293T cells to produce the LV vectors, which enabled the coexpression of the Bdnf gene and the enhanced green fluorescent protein (Egfp) gene. rMSCs were separated from the bone marrow of 2 month-old F344 rats, cultured in vitro, and identified. rMSCs were infected by the LV vectors that were produced already and were identified with fluorescent microscope, RT-PCR, immunocytochemical staining, and western blot. The result of sequencing showed that the sequence of the cloned Bdnf gene was consistent with that reported in the GenBank. The PNL-BDNF-IRES2-EGFP plasmid that was identified showed the correct sequence. After the 3 plasmids of LV vectors were cotransfected to the 293T cells, considerable green fluorescence in 293T cells was observed under the fluorescent microscope; the supernatant was collected and concentrated using ultracentrifugation, and the titer of the replication defective LV vector particles measured was found to be 6.7 x 10(7) TU/mL. After the constructed LV vectors infected the rMSCs, the results obtained using RT-PCR, immunocytochemical staining, and western blot showed that the expression of BDNF in the Bdnf-rMSCs group (experimental group, EG) was significantly higher than that in the PNL-IRES2-EGFP-rMSCs group (mock group, MG) and the rMSCs group (control group, CG) at both mRNA and protein levels. LV vectors carrying the Bdnf gene were constructed successfully. The Bdnf gene-modified rMSCs could express BDNF to a higher degree. This greatly facilitates the next step in the study, such as the long period of therapeutic observation of cerebral ischemia with Bdnf gene-modified rMSCs. PMID- 17460895 TI - [Construction of vector of multiple loci gene targeting in leghorn chicken based on BAC with Cre/lox P system]. AB - Based on the sequence of BAC (Bacterial Artificial Chromosome) along with the Cre/lox P system, the gene-targeting vectors to multiple loci of the repetitive internal transcribed spacers between rDNA genes in Leghorn chicken were constructed. The key material of multiple loci gene targeting in vivo would be obtained. First, the plasmid of pYLSV-TDN with TK, HRDS2, and Neo genes was constructed. The TK-HRDS2-Neo DNA fragment obtained from the plasmid of pYLSV-TDN was digested by Not I/HindIII and inserted into the upstream of the lox P site of BAC plasmid for obtaining the selective vector of BAC-TDN. The expression vector of pYLVS-GID with EGFP, hIFN genes, and HRDS1 was then obtained. The plasmid of BAC-TDN-VS-GID was obtained by cotransformation of the selective vector of BAC TDN and the expression vector of pYLVS-GID to E. coli NS3529 through the action of Cre/lox P system. The gene-targeting vector of BAC-TDN-GID to multiple loci of the ITS region in Leghorn chicken was obtained by cleaving the sequence of pYLVS with the homing endonuclease of I -Sce I and ligating with the linker of LS. The insertion and the insert direction of DNA fragments were identified by restriction digestion or PCR and sequencing in each clone. The significance of the technique ofgene-targeting vector to multiple loci are shown as follows. First, the targeting loci were increased to 100 - 300. Second, the problems of unstable expression of inserted genes were partially solved. Third, the need for safety against toxicity integration was resolved. Fourth, the forbidden zone of gene integrating on the repetitive DNA sequences was broken through. PMID- 17460896 TI - [Isolation, purification and identification of epithelial cells derived from fetal islet-like cell clusters]. AB - The aim of this article is to provide methods for the isolation and identification of pancreatic stem cells and cell source for research and therapy of diabetes. ICCs were isolated by collagenase IV digesting and then cultured; epithelial cells were purified from monolayer cultured ICCs. The growth curve of the epithelial cells was measured by MTT. The expression of molecular markers in the cells was identified by immunohistochemical staining. The surface markers in the epithelial cells were analyzed by FACS. Epithelial cells were purified from isolated human fetal ICCs and passaged 40 times, and 10(6) - 10(8) cells were cryopreservated per passage. The growth curve demonstrated that the epithelial cells proliferated rapidly. The epithelial cells expressed PDX-1, PCNA, CK-7, CK 19, Nestin, Glut2, and Vimentin, but Insulin was undetected. The cells expressed CD29, CD44, and CD166, but did not express CD11a, CD14, CD34, CD45, CD90, CD105, and CD117. Taken together, these results indicate that self-renewable epithelial cells can be isolated and purified from human fetal pancreas. These also show that the epithelial cells originate from ducts and have the characteristics of pancreatic stem cells. PMID- 17460897 TI - [Human bone marrow mesenchymal stem cells differentiated into dopaminergenic neurons in vitro]. AB - Midbrain dopamine (DA) neurons play an essential role in modulating motor control. Defects in central DA neurons affect a wide range of neurological disorders including Parkinson's disease (PD). The greatest motivation in the field has been the potential use of DA neurons for cell transplantation therapy in Parkinsonian patients. Recent studies indicated that BMSCs could differentiate into DA neurons in vitro as neural stem cells (NSC) and embryonic stem cells (ESC) could. However, there are no direct evidences about functional DA neurons derived from BMSCs. According to the protocols which had been applicated in inducing neuronal stem cells and embryonic stem cells differentiate into DA neurons in vitro, the present study provides a protocol by using 50 micromol/L brain derived neurotrophy factor (BDNF), 10 micromol/L forskolin (FSK) and 10 micromol/L dopamine (DA) to induce BMSCs differentiate into DA neurons. After 2 weeks of differentiation, the cells expressed the character of neurons in ultrastructure. RT-PCR discovered mRNA of NSE (neuron specific enolase), Nurr1, Ptx3, Lmx1b and Tyrosine hydroxylase (TH) were positive. Immunocytochemistry staining indicated the ratio of TH-positive neural cells was significantly increased after induced 2 weeks (24.80 +/- 3.36) % compared to that of induction of 3 days (3.77 +/- 1.77) %. And the DA release was also different between differentiated and undifferentiated cells detected by high performance liquid chromatography (HPLC). That is to say BDNF and FSK and DA can induce BMSCs differentiate into DA neurons in vitro, and the transdifferentiated cells express mature neurons characters. BMSCs might be a suitable and available source for the in vitro derivation of DA neurons and cell transplantation therapy in some central neural system diseases such as PD. PMID- 17460898 TI - Antioxidant activities of some local bangladeshi fruits (Artocarpus heterophyllus, Annona squamosa, Terminalia bellirica, Syzygium samarangense, Averrhoa carambola and Olea europa). AB - In the present study, antioxidant activities of the fruits of A. heterophyllus, A. squamosa, T. bellirica, S. samarangense, A. carambola and O. europa were investigated. For this, at first matured fruits of them were sliced into small pieces and dried in the sun and finally crushed in a grinder to make powder. Ethanolic extracts of fruit powder were prepared using 99.99% ethanol. The antioxidative activities of these extracts were determined according to their abilities of scavenging 1, 1-diphenyl-2-picrylhydrazyl (DPPH) free radical. It was demonstrated that all the ethanolic extracts of A. heterophyllus, A. squamosa, T. bellirica, S. samarangense, A. caranbola and O. europa showed antioxidant activities. The IC50 of the ethanolic extracts of A. heterophyllus, A. squamosa, T. bellirica, S. samarangense, A. carambola and O. europa were 410, 250, 34, 200, 30 and 76 microg/mL, respectively. Among them, A. carambola showed the highest antioxidant activities followed by T. bellirica, O. europa, S. samarangense, A. squamosa and A. heterophyllus indicating that fruits of A. carambola, T. bellirica and O. europa are very beneficial to human health. PMID- 17460899 TI - [Proliferation and differentiation of MC 3T3-E1 cells cultured on nanohydroxyapatite/chitosan composite scaffolds]. AB - with chitosan in situ using a chemical method and a porous structure obtained was then lyophilized. Preosteoblast MC 3T3-E1 the scaffolds was examined after staining it with Wright's stain. Their proliferation was assessed using MTZ assay. After being Abstract Nanohydroxyapatite/chitosan composite scaffolds were fabricated and the proliferation and differentiation of preosteoblast MC 3T3-E1 on them were examined for the assessment of their biocompatibility. Nanohydroxyapatite was combined with chitosan in situ using a chemical method and a porous structure obtained was then lyophilized. Preosteoblast MC 333-E1 cells were inoculated into the porous composite scaffolds and chitosan scaffolds, respectively. The morphology of cells cultured on the scaffolds was examined after staining it with Wright's stain. Their proliferation was assessed using MTT assay. After being cultured in conditioned medium for 30 days, the cells' alkaline phosphatase activities on the scaffolds were studied in situ to compare their differentiation levelabout. Moreover, the alkaline phosphatase activities were assessed with a kit. The expression level of characteristic osteogenic gene was evaluated using Reverse Transcription-Polymerase Chain Reaction (RT-PCR). The results indicated that MC 3T3-E1 cells grown on the composite scaffolds showed a higher proliferation rate and spread better than that on chitosan scaffolds. The alkaline phosphatase stain results showed that the alkaline phosphatase activity of cells on composite scaffolds was significantly higher than that on the chitosan scaffolds. In addition, the quantitative examination of alkaline phosphatase activity indicated that the cells cultured on the composite scaffolds expressed an activity level about 8 times higher than that on chitosan scaffolds. Simultaneously, the osteogenic gene osteopontin (OPN) of cells cultured on composite scaffolds showed a higher expression level than that on chitosan scaffolds. Another osteogenic gene osteocalcin (OC) was expressed in cells cultured on composite scaffolds, whereas it was not detected in cells on chitosan scaffolds. The addition of nanohydroxyapatite in the scaffolds improved not only the proliferation but also the differentiation of preosteoblast cultured on them. The composite scaffolds showed good biocompatibility and bioactivity. These scaffolds would be promising in bone tissue engineering. PMID- 17460900 TI - [Purification and characterization of carbonyl enantioselective reductase from Morganella morganii J-8]. AB - The purification and the characteristics of an enzyme from Morganella morganii J 8, which could produce d-pseudoephedrine from 1-phenyl-2-methylamine-acetone, were performed in this study. In this research, first, cells were disrupted by ultrasonic treatment at 4 degrees C. The carbonyl enantioselective reductase was purified with a combination of ammonium precipitation, Phenyl Superose hydrophobic chromatography, DEAE anion exchange, and native polyacrylamide gel electrophoresis. The molecular mass of the purified enzyme subunit was estimated to be 42.5kD on sodium dodecyl sulfate-polyacrylamide electrophoresis (SDS-PAGE). The native molecular mass of the enzyme that was analyzed by high-performance liquid chromatography was found out to be 84.1 kD, which indicated that the enzyme was a dimmer. The purified enzyme was analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, and the result showed that the purified enzyme had high homology with leucine dehydrogenase. PMID- 17460901 TI - [Mutation research on Q23L and Q23LG272E in phytase derivated from Aspergillus fumigatus]. AB - Aspergillus fumigatus wild-type phytase has many favorable properties, such as a good thermorstability and a broad pH optimum. However, the specific activity of the enzyme is relative low. A. fumigatus Q23L phytase resulted in a remarkable increase in specific activity around pH4.5 - 7.0, but the pH stability of Q23L was lower than A. fumigatus wild-type phytase. To increase the pH stability of Q23L, the mutant Q23LG272E was constructed by site-directed mutagenesis with PCR. The gene of A. fumigatus wild-type phytase and the mutant genes encoding the Q23LG272E and the Q23L were correctly expressed in Pichia pastoris GS115. Enzymes were purified and their enzymatic properties were determined. The results revealed that the specific activity of the Q23L improved remarkably, which increased from 51 u/mg of the wild type to 109 u/mg at pH5.5. Meanwhile, the pH stability of Q23L, decreased evidently, especially from pH3.0 to pH4.0.The pH stability of Q23LG272E in pH3.0 - 4.5 and pH6.5 - 7.0 has been improved compared with Q23L. The specific activity of Q23LG272E basically maintained at the level of Q23L. Analysis of 3-D structure and sequence similarity were used to reveal the presumable factors influencing the enzymatic properties of Q23LG272E, and discussion for the relationship between structure and function of phytase was given. PMID- 17460902 TI - [Expression, purification and enzymatic characterization of Thermus thermophilus HB8 aspartate aminotransferase in Escherichia coli]. AB - To obtain thermostable aspartate aminotransferase, the gene aspC from an extremely thermophilic bacterium, Thermus thermophilus HB8 was cloned, and its product was overexpressed in Escherichia coli BL21 (DE3) and Rosetta (DE3). The expression in Rosetta (DP3) was more efficient. The optimum reactive pH was 7, and the recombinant enzyme activity changed little when incubated in the buffer of pH8 - 10 on 37 degrees C for 1 h. The optimum reactive temprature was 75 degrees C, and the recombinant enzyme was more stable on the temperature of 25 - 55 degrees C. The half life of recombinant enzyme on 65 degrees C was 3.5 h, on 75 degrees C was 2.5 h. KmKG was 7.559 mmol/L, VmaxKG was 0.086 mmol/(L x min), KmAsp was 2.031 mmol/L, VmaxAsp was 0.024 mmol/(L x min). Ca2+, Fe3+, Mn2+ inhibited enzyme activity softly. PMID- 17460903 TI - [Improved expression of HLA-A* 2402-BSP in Escherichia coli and its tetramer preparation]. AB - HLA-A* 2402 is one of the most frequently encountered HLA-A alleles in East Asian populations. In order to study the CD8+ T cell responses in Chinese populations, we have described the generation and functional test of HLA-A* 2402 tetramer loaded with HCMV pp65(341-349) peptide (QYDPVAALF, QYD). The cDNA of HLA-A* 2402 heavy chain was cloned by RT-PCR from one of the donors. DNA fragment encoding the ectodomain of HLA-A* 2402 heavy chain fused at its carboxyl-terminal a BirA substrate peptide (BSP) was amplified by PCR with the cloned heavy chain cDNA as a template. The wild-type gene of HLA-A* 2402-BSP was not expressed in Escherichia coli (E. coli), while mutant HLA-A* 2402-BSP gene with optimized codons was overexpressed as inclusion bodies in E. coli. Furthermore, the soluble HLA-A* 2402-QYD monomers were generated by in vitro refolding of washed inclusion bodies in the presence of beta2-microglobulin and QYD peptide. The tetramer was subsequently formed by mixing HLA-A* 2402-QYD monomers with streptavidin-PE at a molar ratio of 4:1. Flow cytometry analysis indicated that this tetramer possessed binding activity with specific CTL from HLA-A24+ donors and the frequencies of tetramer-binding CTL were 0.09% - 0.37% within total CD8+ T cells. This tetrameric agent provides a powerful tool to explore the secrets of CTL responses against HCMV antigens in HLA-A* 2402 individuals. PMID- 17460904 TI - [Preparation and identification of a single-chain antibody fragment against high pathogenic H5N1 avian influenza virus]. AB - Previously, an mAb 10F7 was developed against H5N1 hemagglutinin, which was highly specific to 34 different H5N1 strains and showed good neutralizing activity. In the present study, the single-chain fragment of the antibody was cloned into a prokaryotic vector and then expressed in E. coli. The activity of the scFv was tested in hemagglution inhibition and neutralization experiment. Two H5N1 virus strains were inhibited to bind erythrocyte cells by the scFv while the H9 virus was not. Also, five H5N1 virus strains were neutralized during infecting MDCK cells. These results showed an approachable method for developing therapeutic antibody to H5N1 virus. PMID- 17460905 TI - [Subcellular localization and identification of hydrogenase isolated from the marine green alga Platymonas subcordiformis using immunoprecipitation and MALDI TOF MS]. AB - A marine unicellular green alga, Platymonas subcordiformis, was demonstrated to photobiologically produce hydrogen gas from seawater. The objective of this study was to localize and identify the hydrogenase isolated from P. subcordiformis. Adaptation in the presence of inhibitors of protein biosynthesis indicated that the hydrogenase was much more inhibited by cycloheximide than that by chloramphenicol. The result suggested that the hydrogenase isolated from P. subcordiformis is probably synthesized in cytoplasmic ribosomes. Both Western blot analysis and immunogold electron microscopy demonstrate that the P. subcordiformis hydrogenase is mainly located in the chloroplast stroma. The proteins that reacted specifically with the antibodies against the iron hydrogenase isolated from Chlamydomonas reinhardtii were concentrated by immunoprecipitation. The separated protein bands were cut out of the SDS-PAGE gel, in-gel digested by trypsin, and analyzed by Matrix-Assisted Laser Desorption Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF MS). Mascot was employed for analysis of the MALDI data using the public databases NCBInr. The hydrogenase isolated from P. subcordiformis was identified to be the Fe-hydrogenase. PMID- 17460906 TI - [Effect of the in vitro culture and cryopreservation on the growth of the microencapsulated recombinant cell and endostatin production]. AB - Microencapsulated recombinant cells technology is a novel approach to tumors therapy. It is necessary to prepare a plenty of the microcapsules with better cell viability and higher endostatin production in order to bring this technology into the clinic. The in vitro culture and cryopreservation are very important parameters in the preparation of microencapsulated cells. In this work, we studied the effect of the in vitro culture and cryopreservation on microencapsulated recombinant cells growth and endostatin production and the effect of the in vitro culture on the cryopreservation of microencapsulated recombinant cells. The results showed that the time of in vitro culture potently affected microencapsulated recombinant CHO cells growth in vivo, endostatin production and the microcapsule stability. The microcapsule kept intact after 36 days of implantation when the in vitro culture time was under 4 days. The thawed microencapsulated recombinant CHO cells had better cell growth and higher endostatin production after 40 days of cryopreservation when the in vitro culture time was 4 days and 8 days. Therefore, the best in vitro culture time was 4 days according to the results of the in vivo culture and cryopreservation and the cryopreservation did not affect microencapsulated recombinant CHO cells growth in vivo, endostatin production and the microcapsule stability. PMID- 17460907 TI - [Priming with an HEV Th epitope can improve the humoral immunogenicity of its native protein]. AB - A dominant H-2d restricted Th epitope P34 was found to be contained in recombinant particulate hepatitis E virus (HEV) vaccine HEV 239. In this paper, the cellular immune response induced in P34 immunized BALB/c mice were studied and the priming effect of P34 was characterized. Groups of BALB/c mice were subcutaneously (s. c.) immunized with P34, splenocytes were then stimulated with P34 and HEV 239 protein, cellular immune response was assayed by IFN-gamma ELISPOT, flow cytometry and T cell proliferation experiments. Results showed that P34 immunized BALB/c splenocytes responsed to P34 and HEV 239 protein stimulation in IFN-gamma-ELISPOT, flow cytometry and T cell proliferation experiments. After depletion of the CD4+ T cells from the immunized splenocytes by magnetic separation, the response decreased to the background level while almost no influence was observed after CD8 + T cells depletion which showed that the cells responsible for IFN-gamma secretion were mainly CD4+ T cells. Then mice were primed with P34 and boosted with its vector protein, E2, the E2 specific antibody titer were assayed. Results showed that after P34 priming, some of the 10 microg, 20 microg E2 boosted mice could develop anti-E2 antibody 1 week later and all the mice had detectable antibody 3 weeks after boosting. In the control peptide P18 priming group, even after boosting with 20 microg E2, anti-E2 antibody couldn't be detected until the end of this experiment. The results showed that priming with P34 epitope could increase the immunogenicity of its vector protein, E2, in BALB/c mice. PMID- 17460908 TI - [The surface display of porcine parvovirus VP2 protein in Lactobacillus casei]. AB - Lactobacillus casei 393 was selected as a bacterial carrier for the expression of Porcine Parvovirus (PPV) protective antigen VP2 protein. The gene encoding PPV VP2 protein was cloned into the Lactobacillus casei surface expression vector pPG, and then the constructed recombinant vector pPG-VP2 was electrotransformed into Lactobacillus casei 393 generating the recombinant system pPG-VP2/L. casei393 expressing PPV VP2 protein. The recombinant strain was induced by 2% Lactose in MRS and about 74kD protein was detected with SDS-PAGE. The result of Western-blot indicated that the expressed protein possessed the antigenic specificity which could be recognized by mouse anti-PPV serum. The indirect immunofluorescent test showed that the expressed protein was secreted on the cell surface Lactobacillus casei. PMID- 17460909 TI - [Construction of a transfer vector based on canine adenovirus type-2]. AB - Canine adenovirus type 2 (CAV-2) has been proposed as a vector for recombinant vaccine. Alternatively, it may be an attractive tool for gene transfer due to lack of pre-existing immunity in humans. In this study, a transfer vector based on CAV-2, in which the 1381bp fragment of the E3 region was deleted, and a linker containing the Not I, Cla I, Fse I restriction enzyme sites were cloned into the deleted region. The recombinant CAV-2 genome was released from the plasmids enzyme digestion and transfected into MDCK cells by lipofectamine to obtain the recombinant virus. No significant difference in morphology, hemagglutination and replication between the recombinant and the wide type CAV-2 was found. These results indicated that this recombinant virus CAV-2-deltaE3 (NF) may be an efficient vector for gene transfer and the capacity of the vector for inserted foreign gene was up to 3.3kb. PMID- 17460910 TI - [Study on CTP production from CMP by beer yeast cell immobilized in PVA]. AB - With PVA as the carrier, the frozen beer yeast cells were immobilized for production of CTP from CMP. we explored the optimal condition of the immobilization from the aspects of the type, concentration of the PVA, and the immobilizing methods of cells In all 8 continuous batch of fermentation under the reactional condition of the immobilized cells, the conversion rate of CTP were maintained about 85% - 95%. Moreever, the storage stability of immobilized cells were investigated, and the products was also isolated and identifided by HPLC. PMID- 17460911 TI - [Effect of spermine on cell growth and polysaccharide production in suspension cultures of protocorm-like bodies from Dendrobium huoshanense]. AB - The effect of outer spermine on cell growth, accumulation of polysaccharides and utilization of nutrient together with the intracellular polyamine contents were investigated in suspension cultures of protocorm-like bodies from Dendrobium huoshanense. The results indicated that spermine at 0.6 mmol/L was the most effective in increasing cell growth and polysaccharide synthesis. The specific growth rate of cell increased from 0.046d(-1) to 0.054d(-1), and the maximum dry weight and polysaccharide production reached 32.4g DW/L and 2.46g/L respectively, which were 1.32-fold and 1.31-fold that of the control on day 30. The titres of intracellular free polyamines were higher in the cultures treated with spermine than that of the control. Invertase and nitrate reductase activities were found to increase significantly in the cultured cells treated with spermine, which was beneficial to the utilization of carbon and nitrogen source. PMID- 17460912 TI - [Simulation and analysis of ethanol concentration response to enzyme amount changes in Saccharomyces cerevisiae glycolysis pathway model]. AB - Metabolome has become an important part of Systems Biology, and a large set of data has already gained by applying the methods of metabolome. How to deal with the data and how to combine data of metabolome with data of other omics are problems that can not be ignored. An Enzyme Amount Multiple Factor was imported into the enzyme kinetic equation. When the enzyme amount in the system changed, in silico model, it means to alter the Enzyme Amount Multiple Factor. In order to observe ethanol concentration response to enzyme amount changes in S. cerevisiae glycolysis pathway model, enzyme amount was separately set at high and low level, the corresponding Enzyme Amount Multiple Factor value was 10 and 0.1, relatively. Based on the result of simulation, twelve enzymes in pathway were separated into two classes, class I and class II by cluster analysis. The four enzymes belonging to class I, ADH, HK, PFK and PDC, all catalyze irreversible reactions. The six out of eight enzymes belonging to class II, ALD, GAPDH, GlcTrans, lpPEP, PGI and TIM, catalyze reversible reactions. The other two enzymes belonging to class II, lpGlyc and PK, catalyze irreversible reactions. Based on this method, data of metabolome and proteomics are easily integrated to accomplish relatively overall analysis of system properties. PMID- 17460913 TI - [Effect of polyethylene glycol on the regeneration of the callus of Pogonatherum paniceum under different culture conditions]. AB - This paper studied the effect of polyethylene glycol (PEG) on regeneration and free proline accumulation of callus of Pogonatherum paniceum (Lam.) Hack. under motionless liquid culture condition and shake liquid culture condition. Callus of P. paniceum had the ability to resist the stress of PEG. The effects of PEG stress and culture conditions on the callus of P. paniceum appeared mainly in two aspects, delaying regeneration time and debasing regeneration rates. The shake liquid culture mainly delayed the regeneration time and PEG stress mainly debased the regeneration rates. Free proline accumulated in the two culture conditions, and the contents of proline were positively correlated with PEG concentrations and culture time. After stress removal, most of the callus could recover the ability of regeneration, and the free proline might pay an important part in the inhibition and recovery. So it must be chosen a more than 300 g x L(-1) PEG concentration and long than 3 weeks culture time in the selection of drought resistant mutants of P. paniceum. The motionless liquid culture was more suitable for selection of drought-resistant mutants. PMID- 17460914 TI - [Establishment of a cell-based high-throughput screening model for PPARdelta agonists]. AB - To establish a new high-throughput screening model for the agonist of PPARdelta, PPARdelta gene was obtained by reverse transcriptase-polymerase chain reaction (RT-PCR), and subcloned to pGEM-T Vector for sequencing, then the PPARdelta fragment was excised by restriction enzymes, and inserted into pTARGET Vector to construct expression vector pTARGET-ppARdelta. Insert three copies of PPRE into pGl3-promoter vector to construct expression vector pGl3-PPRE x 3-luc. The vector pTARGET-ppARdelta was transiently cotransfected with pGl3-PPRE x 3-luc into different cell lines to assay the expression levels of luciferase. The PPARdelta agonist screening model was established and optimized. Bezafibrate and linoleic acid can induce the expression of luciferase significantly and in a dose dependent manner. This method can be used for high throughput screening for the agonist of PPARdelta, which might become lead compounds for new anti atheroscleriosis or anti-adiposity drugs. PMID- 17460915 TI - [The study of optimal conditions of electroporation in Escherichia coli DH10B strain]. AB - In order to optimize the conditions of construction BAC library, the transformation efficiency of E. coli DH10B was studied in this paper. Our data prove much higher competence of electroporation (reaches 2.19 x 10(10) cfu/microg pUC19 DNA) when harvesting the cells between an OD550 of 0.7 - 0.8. Five different electric field strength (from 9 kV/cm to 25 kV/cm) and three different sized plasmid vector DNAs including pUC19 DNA, pECBAC1 DNA and pCLD04541 DNA, as well as three bacterial artificial chromosomes (BACs) ranging from 40 to 190 kb and their mixture were used to discover the transformation efficiency changes under various conditions. Our data show maximum transformation efficiency and optimal electric field strength of plasmid DNAs drop dramatically with increasing size of the DNA. Molecules of 190 kb transform more than 50-fold less well, on a molar basis, than molecules of 40 kb. And the optimal voltage gradient is strongly dependent on the different sized molecules, for instance, pUC19 reaches the highest transformation efficiency at 21 kV/cm, while the 180 kb BAC DNA gets its best efficiency at 13 kV/cm. This paper demonstrates that conditions may be selected which increase the average size of BAC clones generated by electroporation and could be widely applied in large-insert genome library construction. PMID- 17460916 TI - [Cloning and expression in Escherichia coli of secondary lymphoid-tissue chemokine (SLC) gene]. AB - Secondary lymphoid-tissue chemokine (SLC) is a type of CC chemokine identified by searching the Expressed Sequence Tag (EST) database. The full-length SLC gene was synthesized based on human SLC sequence using SOE-PCR. The sequenced SLC gene was cloned into expression vector pTMF and pALM, which used to transform Escherichia coli. Then the E. coli was cultured and induced according to protocol. The expressed target protein was identified by Western blotting. The target protein was expressed as soluble protein as well as inclusion bodies, the ratio of these two forms target protein varied with the difference conditions of culture and induction. The target protein was purified with the methods of nickel nitrilotriacetic acid (Ni-NTA) metal-affinity chromatography. The results of electrophoresis of the purified target protein showed that the molecular weight was larger than the predicted molecular weight. PMID- 17460917 TI - [Transformation of Dunaliella salina by using glass beads--a novel transformation method]. AB - A novel transformation method was firstly established using glass beads in Dunaliella salina (D. salina). The results showed that the GUS gene, a reporter gene, was successfully expressed in D. salina. Cells of D. salina presented blue color under the microscope after stained. In addition, different factors which influenced transformation were optimized including the transformation consecutive time, rotate speed, concentration of the plasmid and PEG 6000. The experiment indicated that this fit together can obtain the best results for D. salina transformation: adding 150 microL PEG and 90 microL plasmid DNA to 800 microL culture of D. salina (10(6) cells/mL) containing 300 mg glass beads, swirling 12 seconds under the rotate speed 2400r/min. This newly method can be used as a potential tool in the research of D. salina gene engineering with the advantage of more simpleness, convenience, quickness and less expense. PMID- 17460918 TI - [Tuberculosis in a pair of twins--the use of molecular biology methods for the detection of the source of infection]. AB - Tuberculosis is still one of the most prevalent infectious diseases worldwide with a high annual morbidity and mortality rate. Its mode of spread necessitates prompt investigation to identify those with active disease, possible carriers and as many contacts as possible. This article describes a pair of twins with endobronchial tuberculosis following close contact with a family relative who had active disease. Mycobacterium tuberculosis was isolated from the family relative and the two children. Laboratory diagnosis of the mycobacterial strain and epidemiologic follow-up were performed using a molecular biology tool, the restriction fragment length polymorphism method. Treatment was successful due to the close cooperation between the medical staff of the hospitals, the community clinics and the Center for Diagnosis and Treatment of Tuberculosis. Treatment was administered by the directly observed therapy (DOT) method recommended by the World Health Organization. This article describes the clinical course and treatment of the patients and reviews the new molecular biology methods currently being used for the diagnosis of tuberculosis and their important clinical applications. PMID- 17460919 TI - [Open versus laparoscopic appendectomy in children in a regional hospital in Israel]. AB - Although laparoscopic appendectomy (LA) has already been found to be associated with greater diagnostic accuracy, less post-operative pain and shorter hospital stay as compared to open appendectomy (OA), questions remain regarding the advantages of this approach and it is still not widely practiced in children, especially in regional hospitals. AIM: This study aims to evaluate Ha'emek Medical Center's initial experience with pediatric OA and LA between July 2002 and October 2003. METHODS: This study is a retrospective outcome analysis of pediatric OA and LA. Records of all children aged 0 to 14 years who underwent appendectomy for acute appendicitis, were reviewed. Operating time (OT), antibiotic treatment, analgesic needs, length of stay (LOS) and complications were analyzed and were compared between the groups. RESULTS: Seventy-one children with acute appendicitis underwent appendectomy in Ha'emek Medical Center between July 2002 and October 2003. Fifty-two children underwent OA and 19 underwent LA. There were no differences in age, sex, race, or stage of the appendicitis between the groups. LA took longer than OA (68 vs 37 minutes), length of stay was shorter for LA (2.8 vs 4.3 days), complications rate was not significantly different and overall hospital expenses were less for LA (NIS 5,756.95 vs. NIS 6,055.47). CONCLUSION: LA is as safe as OA and although it takes longer, recovery is faster. We, therefore, conclude that LA can be safely recommended for treating children with acute non-perforated appendicitis. PMID- 17460920 TI - [Laparoscopic colon and rectal surgery--after ten years and 350 operations]. AB - BACKGROUND: Within a decade since laparoscopy was used in cholecystectomy it has become the preferred approach in many abdominal procedures. Laparoscopic colon and rectal surgery has not yet been adopted by the majority of surgeons, due to technical complexity and reservation regarding its oncological safety. As data and experience accumulate, this attitude is gradually changing. We present our experience with laparoscopic surgery of the large bowel over the last ten years. AIM: To assess the short and intermediate term results after laparoscopic colon and rectal surgery, and to summarize the long term results after curative colectomy for malignancy. METHODS: Data regarding all patients undergoing laparoscopic colon and rectal surgery was prospectively entered into a computerized database, including demographics, surgical technique and perioperative course. Follow-up information was gathered at outpatient clinic visits, and using telephone interviews in selected cases. Data analysis was performed using a statistical software package. RESULTS: Over a period of ten years, 350 various laparoscopic colon and rectal procedures were performed, for both benign and malignant conditions. Sixty percent of the operations were for treatment of colorectal cancer. In 14.5% of cases conversion to open laparotomy was required. Post-operative complications included surgical site infection in 17.4%, anastomotic leak in 6.9%, and a mortality rate of 2.8%. Long term follow up revealed cancer recurrence locally in 2.3% and systemically in 8.2%. Five year survival was 56% after resection of colorectal cancer regardless of the stage, and 63% after resection with curative intent. CONCLUSIONS: The laparoscopic approach to large bowel surgery enables short and long term results comparable with those achieved by open technique, regarding perioperative complication rate and long term oncologic outcome. The advantages of laparoscopy, related to reduced abdominal wall trauma, justify the adoption of this technique as a legitimate alternative to the open approach. PMID- 17460921 TI - [Biphasic versus monophasic shock waveforms for transthoracic cardioversion of atrial flutter in the emergency room]. AB - Transthoracic electrical cardioversion (ECV), traditionally using monophasic waveform (MW) shock, has an important role in the treatment of symptomatic atrial flutter (AFI). Biphasic waveform (BW) shock has been demonstrated to be more successful than MW shock for termination of atrial fibrillation, but data about its use for ECV of AFI are limited. METHODS AND RESULTS: We retrospectively analyzed the records of 53 patients (pts) admitted -to the ER due to symptomatic AFl during the period August 2004 to August 2005: 31 pts received BW shock and 22 pts MW shock. The type of shock waveforms and the initial energy of CV were chosen by the doctor on duty in the ER; the lower energy for ECV was 20 joules, which was increased to 50, 100 and 200 joules if necessary. There were no significant differences between the clinical characteristics of the pts who received BW shock or MW shock. All pts underwent ECV via anterior-laterally positioned hand-held electrode paddles. Successful ECV by BW shock and MW shock was 41% and 42% of the pts, respectively, using 20 joules of energy (p=n.s.); 77% and 80% using 50 joules (p=n.s.); 93% and 90% using 100 joules (p = n.s.); 100% of successful ECV was reached when 200 joules of energy was used, regardless of waveforms type. Median energy for successful ECV was 50 joules in both types of electrical waveforms. No complications were reported. CONCLUSIONS: There were no significant differences in the success rates of conversion of atrial flutter to sinus rhythm by BW or MW shock. We recommend 50 joules for starting energy of ECV of AF1 regardless of waveforms type. PMID- 17460922 TI - [Cigarette use among Israeli adolescents: why do they smoke?]. AB - BACKGROUND: Efforts to prevent adolescents from smoking have limited efficacy, with little understanding of why they smoke. We examine many of the factors which may influence a teenager's decision to start smoking. METHODS: A self-completed questionnaire was distributed among pre-draft teenagers (age 16-18 years) with questions regarding current smoking habits as well as smoking habits of family members. Smokers were asked why they started, and attitudes regarding the dangers of smoking and anti-smoking legislation were examined. RESULTS: A total of 1067 questionnaires were completed (707 males, 360 females). In all, 25.7% were current smokers (26.2% of male and 21.1% of females; p=0.069), with smoking rates highest among those born in Middle Eastern countries and lowest among those born in Ethiopia. Those from single-parent families had a higher smoking rate (relative risk: 1.28), as did those with either parents or siblings who smoked. Parents who were smokers were more likely to tell their children not to smoke, while smoking teenagers were more likely to have been told by their parents not to smoke. Most smokers (58.2%) did not give a specific reason for smoking while 30.3% listed "peer pressure", 8.4% "parents smoking" and 3.1% "advertisements" as the major factor. Smokers were less likely to agree that smoking is harmful and more likely to oppose anti-smoking legislative measures. CONCLUSIONS: Many factors influence a teenager's decision to smoke, including cultural norms and habits of friends and family members. Educating both adolescents and those around them may help prevent smoking among this age group. PMID- 17460923 TI - [Pediatric endourology]. AB - OBJECTIVE: To assess the outcome of pediatric patients treated by an endourological approach for various urinary tract pathologies. METHODS AND MATERIALS: Thirty-seven children (median age 5 years, range 0.3-14 years) were endoscopically treated for ureteropelvic junction obstruction (UPJO) (n= 6), ureteral strictures (n=5), upper urinary tract calculi (n=21) and bladder calculi (n = 5). RESULT: Upper urinary tract calculi were approached by ureteroscopy (n=12), retrograde intrarenal surgery (n=6) and percutaneous nephrolithotomy (n = 3). The average stone burden was 11 mm (range 5-35 mm) and operative time was 40 minutes (range 15-120 minutes). Bladder calculi were treated percutaneously in 3 cases and transurethrally in 2 cases for an average stone burden of 34 mm (range 7-120 mm). The overall stone-free rate after one procedure was 96%. UPJOs were retrogradely approached in an average operative time of 40 minutes (range 30-50 minutes). Successful clinical and functional outcome was maintained after an average follow-up of 15 months (range 6-30 months). The 5 ureteral strictures included 2 located in the middle ureter and 3 at the ureterovesical junction. The success rate in this group was 80% and the average follow-up 24 months (range 6 40 months). The median hospitalization time for the entire series was 1 day (range 0-7 days). There were no intraoperative complications. Three (8%) patients developed post-operative urinary tract infections. Delayed anterior urethral stricture occurred in 1 case. No additional complications occurred after an average follow-up of 11 months (range 4-36 months). CONCLUSION: Endourology in children is safe and highly effective. It appears that the indications for endourological treatment in children emulate those of adults. PMID- 17460924 TI - [On health, sense and confidentiality]. AB - Medical confidentiality is a cornerstone of doctor-patient relations. Although physicians are obliged to keep to themselves any information concerning their patients, it is understood that this obligation is not absolute. There are extreme situations in which doctors are expected, by law, to break medical confidentiality for the benefit of the public. One such unusual situation is the case of HIV carriers who refuse to notify their sexual partners concerning their medical condition. This ignorance on the partner's side might endanger his/her life. As there is no way to enforce the HIV carrier to expose his/her secret while he/she is not willing to do so, it is suggested that the disclosure of this sensitive information should be the physician's task. Firstly, the doctor should obtain an ethical committee's approval. It is also suggested that in these rare cases where doctors will have no choice but to reveal information concerning a patient, the issue will be publicized in a proper way, in order to let the public know the circumstances of the particular case. It is hoped that such publicity will be an appropriate answer to any doubts concerning doctors' behavior regarding medical confidentiality. PMID- 17460925 TI - [Laparoscopic colectomy--recommended routinely?]. AB - The current data of laparoscopic versus open colectomy does not enable a definitive conclusion as to whether laparoscopic colectomy should be considered as the procedure of choice and recommended routinely. Laparoscopic colectomy entails a cosmetic advantage, a shorter scar and a limited advantage in short term convalescence. However, laparoscopic colectomy requires a significantly prolonged operation, with possible exposure to more intra-and post-operative complications with a possible conversion to open surgery at the rate of 10%-20% and at a significantly higher cost. It is possible that the cosmetic and borderline quality of life advantages, do no justify the routine utilization of laparoscopic colectomy. Nonetheless, it should be emphasized that laparoscopic colectomy is a legitimate option for patients with benign conditions and uncomplicated colon cancer. The process of assimilation of laparoscopic colectomy should be conducted cautiously, not due to conservatism, but rather taking into consideration the very long learning-curve of this technigue and also from patient care perspectives. Even large, leading referral colorectal centers are usually slow at implementing laparoscopic colectomy, not exceeding 25% of total colon surgery. PMID- 17460926 TI - [Tuberculosis in rheumatic diseases: an ancient enemy in a new era]. PMID- 17460927 TI - [Echinacea--much ado (meanwhile) about nothing?]. PMID- 17460928 TI - [Aids and the right to privacy]. AB - Fear of AIDS exceeds by far the apprehension concerning any other disease: it is mostly irrational, and is driven by a distorted perception of the disease as well as the stigmatization of patients. A major question faced by healthcare systems worldwide concerns the relation between the patients' responsibility to prevent spreading of the virus and the responsibility of the healthcare authorities on this matter. Early HIV detection and treatment, together with proper patients' education, reduce the odds of further infections. Therefore, HIV patients who choose to refrain from partner notification present a significant challenge to the medical staff, as they may rely on human rights and patients' rights principles. This paper deals with dilemmas concerning nonconsensual HIV reporting. The authors advocate a distinction between two types of sexual relationships: whereas in casual relations, both partners share (non-equivalent) responsibility for their safety, in a monogamous and stable relationship, the infected partner is responsible both ethically and legally, for partner notification. Nonconsensual notification is ethically acceptable in the latter type of relationships rather than in the former. Such notification should be as discrete as possible and patients' protection against potential violence and discrimination must be safeguarded. Prior to the notification, the HIV patient should be provided with advance notice. According to the Israeli Patients' Rights Law, such notification is subject to the authorization of a statutory ethics committee. The authors additionally contend that the adherence to the human rights of HIV patients promotes public health interests of preventing the spread of AIDS. PMID- 17460929 TI - [Tuberculosis in the 21st century]. AB - Tuberculosis remains a significant global problem, principally affecting the developing countries, which are also heavily afflicted by HIV. In the industrialized countries, tuberculosis concerns mostly the immigrant, the immuno compromised, and the elderly. More recently, TNF-alpha antagonists are being recognized as an important risk factor for tuberculosis, and guidelines for improved screening and control have been issued. Diagnosis of tuberculosis remains cumbersome, and the newer nucleic acid-based techniques are insufficiently sensitive. New immunologic methods to improve the diagnosis of latent tuberculosis are being studied. Isoniazid and rifampicin remain the mainstay of the treatment of active disease, but several new drugs are being evaluated. The existence of immune reconstitution inflammatory reactions in HIV patients is also becoming more widely appreciated. Treatment of latent tuberculosis still relies mainly on the use of isoniazid, and efforts continue to improve patient compliance. PMID- 17460930 TI - [Tuberculosis among patients treated with TNF-alpha blockers]. AB - Several reports have indicated a relationship between reactivation of latent tuberculosis infection (LTBI) and treatment with tumor necrosis factor alpha (TNF) blocking agents. These biologic drugs have demonstrated high efficacy in the treatment of chronic inflammatory diseases. Several guidelines have been published worldwide suggesting pretreatment screening strategies for latent tuberculosis in candidates for TNF-blocking treatment, as well as a regimen for prophylactic therapy if latent tuberculosis is indeed diagnosed. There are considerable variations among the different guidelines. This review summarizes currently available data on the risk of infection and particularly, the increased risk profile for tuberculosis in patients treated with TNF-blocking agents. The role of TNF alpha blockers in the pathogenesis of tuberculosis and the existing protocols that have been formulated in order to minimize the risk of tuberculosis reactivation during TNF blockade are also outlined. PMID- 17460931 TI - [Azithromycin as a novel treatment for bronchiolitis obliterans syndrome]. AB - Bronchiolitis obliterans syndrome (BOS) is one of the main causes of mortality after lung and bone marrow transplantation. Up to 75% of lung transplantation patients develop BOS within 5 years, whereas after bone marrow transplantation 14% of the patients develop the disease with 65% mortality within 3 years. Patients demonstrate gradual decrease in pulmonary functions with no significant anatomic/imagine findings. Therapeutic trials with high dose systemic corticosteroids, immune suppression and immunomodulation did not show any significant success. However, macrolides and especially azithromycin have recently been reported as highly efficient for BOS. This review summarizes information on the disease focusing on the clinical experience with azithromycin as a treatment for BOS. PMID- 17460932 TI - [Hyperbaric oxygen therapy for chronic diabetic wounds of the lower limbs--a review of the literature]. AB - Chronic wounds of the lower limbs are a cause of severe morbidity in diabetic patients. Low oxygen tension around the wound is one of several critical factors, which mutually enhance the progression of a chronic ulcer. Hyperbaric oxygen therapy (HBO) is believed to improve wound healing by enhancing oxygen tension around the wound. While conventional therapies for diabetic foot ulcer are based on scientific evidence, HBO treatment lacks evidence-based support regarding its cost effectiveness and efficacy. Recently, several publications emerged, which improve our knowledge regarding this subject. This paper briefly reviews the pathophysiology of chronic diabetic ulcers and the possible advantage of HBO therapy in this clinical setting. The article also summarizes the results of relevant publications, in which appropriate scientific measures were applied. In conclusion, there is evidence that HBO therapy reduces the need for major amputations among diabetic patients with chronic ulcers of the lower limb. HBO seems to enhance the rate of healing. Few publications with methodological defects diminish the value of these conclusions. However, there is a need for larger randomized, double blinded studies in order to validate this treatment. PMID- 17460933 TI - [Cyanides--treatment beneath the shade of terror]. AB - Although the use of cyanides as warfare agents has not been documented since the Iran-Iraq war in the 1980s, there are rising fears of cyanide being used by terrorists. An Al-Qaeda terror plot to use cyanide gas in the London Underground was foiled in 2002. The threat of similar events becomes more imminent in light of the terror attacks in our country and worldwide, accompanied by statements and threats by fundamentalist leaders to employ chemical weapons. Therefore, mass intoxication with cyanides is not merely a hypothetical scenario. The treatment of cyanide poisoning is under constant evaluation and there is no international consensus on the subject. The medical treatment of victims at the scene and in hospitals should be rapid and efficient. Current treatment dictates establishing an intravenous line and a slow rate of administration of antidotes. Both demands are not feasible in this specific mass casualty event. The clinical signs of cyanide poisoning are complex, variable and not necessarily obvious for the medical team. There is great interest in reconsidering the existing treatment protocols for cyanide intoxication in light of current research. This review describes the mechanisms of cyanide toxicity, clinical signs of exposure, and current treatment protocols in use worldwide. On the basis of this evidence we suggest a medical treatment protocol for a mass casualty event caused by cyanide. PMID- 17460934 TI - [Guidelines of the Israeli association of rheumatology for the prevention of tuberculosis in patients treated with TNF-alpha blockers]. AB - The use of TNFalpha blockers is associated with reactivation of tuberculosis. The Israeli Association of Rheumatology nominated a committee to determine guidelines for the prevention of tuberculosis in patients taking TNFalpha blockers. The risk of reactivation of tuberculosis is higher with monoclonal antibodies to TNFalpha (infliximab, adalimumab) in comparison with the soluble receptor of TNFalpha (etanercept). All patients who are candidates to receive TNFalpha blockers should be screened for active or latent tuberculosis. The screening includes: Tuberculin Skin Test (TST), chest X-ray and a questionnaire about possible exposure to tuberculosis. Two-step screening should be used as recommended by the Ministry of Health. The reaction elicited in the second test (if applied) should be used. In the general population latent tuberculosis is diagnosed when the TST response is 15 mm. or above; a reaction of 10 mm. or above is positive in populations with a history of definite or probable exposure to TB and 5 mm. is the threshold for populations who are immunosuppressed or if chest radiography reveals old tuberculosis without clear documentation of previous treatment. Patients with a TST less than 5 mm. should be questioned about prior exposure to tuberculosis. Latent tuberculosis should be treated with a 9 month course of isoniazid (300mg/d) or a 4 month course of rifampicin (600mg/d) or for 3 months with a combination of 300 mg. isoniazid and 600 mg. rifampicin daily. The committee recommends postponing treatment with TNFalpha blockers until completion of anti tuberculosis therapy. If the clinical condition requires the urgent use of TNFalpha blockers, these may be initiated one month after starting treatment for latent tuberculosis. PMID- 17460935 TI - [Assessment of the preimaginal stages of the ticks collected from small mammals in Western and Northern Caucasus (Acari: Ixodidae)]. AB - The exact identification of the preimaginal stages of the ticks collected from small mammals in Western and Northern Caucasus as a result of ten-year route investigations allowed us to obtain new data on the geographic ranges, altitude and biotopic preferences, and host-parasite relations of 11 species. A wide range of the joint occurrence (in the same locality, on the same host species, on the same host individual) is recorded for the first time for 10 species. The cohabitation of the each of 10 species with other 2-9 species has been found. The number of the cases of joint occurrence of different ixodid species on the same host individual is proportionate to the collection period in one locality. The cohabitation did not observed only for the species Ixodes ghilarovi, a high mountain ixodid species, which is for the first time recorded in 7 localities of Northern and Western Caucasus. PMID- 17460936 TI - [Morphofunctional changes in the midgut of the Ixodes ricinus nymphs (Acari: Ixodidae) during developmental diapause]. AB - Changes in the midgut of the Ixodes ricinus nymphs at the stage of developmental diapause were studied. It is established, that the midgut of the tick nymphs undergoes the identical and synchronous changes at the development without diapause and in the state of diapause. In 7-8 months after feeding the midgut of the nymphs containes the digestive cells with gematine and food inclusions, as well as reserve cells, like the midgut of unengorged moulted female. But the midgut of diapausing nymphs retaines such condition during the whole period of diapause. PMID- 17460937 TI - [The taxonomic diversity of the parasites of fishes in the Volga basin. III. Aspidogastrea and Trematoda]. AB - A checklist of Aspidogastrea and Trematoda parasitizing fishes in the Volga basin is given. The checklist includes host species names for the each parasite and data on the occurrence of parasites in different parts of the Volga basin. The data on trematodes from 69 fish species are presented. The list containes 107 species and 1 subspecies names of trematodes, and 1 species of Aspidogastrea. The family Diplostomidae is represented by the most number of species (22). Six species of trematodes (Amurotrema dombrovskajae, Sanguinicola skrjabini, Nicolla skrjabini, Plagioporus skrjabini, Apophallus muehlingi, Rossikotrema donicum) have been introduced into Volga River along with aclimatizated fishes or penetrated into its basin by self-migration to new water bodies. PMID- 17460938 TI - [Opisthorchis felineus (Rivolta, 1884) and Metorchis bilis (Braun, 1890) infections in population of some regions of the Ob River Basin]. AB - The incidence of Opisthorchis felineus (Rivolta, 1884) and Metorchis bilis (Braun, 1890) infections among people living in several regions of the Ob River basin in the West Siberia has been assesed in this work. Our results suggest that Metorchis bilis infection was common in many of the serologically tested people. Moreover, this helminth was obtained from the biliary ducts of humans in autopsy. PMID- 17460939 TI - [The effect of the homogenates from different developmental stages of the nematode Protostrongylus rufescens (Leuckart, 1895) on mitochondrial and lipid bilayer membranes]. AB - The effect of the homogenates from different developmental stages of the nematode Protostrongylus rufescens on mitochondrial and lipid bilayer membranes has been studied. The homogenate of P. rufescens affects efficiently the cell energy by the inhibition of the mitochondrial respiration in the metabolic state V3, uncouples oxidative phosphorylation and affects the functions of mitochondria at the level of cyclosporine A-sensitive pore by making it highly permeable. Moreover, the nematode homogenate at the concentration of 1 mkg/ml increases efficiently the integral permeability of lipid bilayer membranes. An increase in this permeability is connected apparently with the formation of single ion channels. The channels of lipid bilayer membranes induced by the nematode homogenate show cation selectivity. PMID- 17460940 TI - [Parasite fauna of young landlocked salmon (Salmo salar m. Sebago girard) in the Pista River (the White Sea Basin)]. AB - A native population of landlocked salmon of the Pista River was investigated in 2000-2002. Adult salmon in Pista River has smallest size among other populations of landlocked salmon in Karelia. Data on the biology and parasite fauna of young salmon are presented. The presence of local salmon populations in lakes of the river system is apparently one of the mechanisms keeping the magnitude of population. The presence of Gyrodactylus salaris, a harmful parasite of the young landlocked salmon, is established in this territory for the first time. This monogenean species is believed to have been introduced into the Pista River via stocking from Finland. PMID- 17460941 TI - [New species of Myxosporidia (Myxosporea: Cnidosporidia) from cottids (Abyssocottidae) and the oilfishes (Comephorus spp.) of Baikal Lake]. AB - Two new species of Myxosporidia, Myxidium baicalensis sp. n., Myxidium donecae sp. n. are described from the gall bladder of fishes from Baikal. Myxidium baicalensis sp. n. parasitising gobies Cyphocottus megalops, Asprocottus platycephalus, and other host species, while Myxidium donecae sp. n. parasitising Baikal oil fishes Comephorus baicalensis and C. dybowski. PMID- 17460942 TI - [First finding of the Centrorhynchus aluconis (Muller, 1780) (Giganthorhynchidae) and Moniliformis moniliformis bremser, 1811 (Moniliformidae) larvae in shrews (Insectivora: Soricidae) of the fauna of Russia]. AB - The larvae of acanthocephalans Centrorhynchus aluconis (Muller, 1780) and Moniliformis moniliformis Bremser, 1811 are recorded for the first time from shrews in Russia (Samarskaya Luka National Park, Samara Region). Taxonomic descriptions and figures of the specimens examined are presented. PMID- 17460943 TI - Basal serum tryptase level correlates with severity of hymenoptera sting and age. AB - BACKGROUND: Increased serum tryptase has been linked to the severity of the reaction after Hymenoptera stings. The aim of the study was to measure basal tryptase levels in patients with Hymenoptera venom allergy and investigate the possible correlation between these levels and the severity of sting reaction. METHODS: One hundred nine patients were included in the study. Sixty-three were wasp venom-allergic and 46 were honey bee venom-allergic. Basal serum tryptase levels were measured by UniCAP. RESULTS: Basal serum tryptase levels were elevated in 12 (11%) of the 109 patients. Levels were 5.14 pg/L (3.62-5.84), 5.3 microg/L (2.94-6.54), 5.18 microg/L (3.71-6.25), and 6.98 microg/L (4.78-12.6), for patients with sting reactions of grade I, II, III and IV (as classified by Mueller), respectively. Basal serum tryptase levels correlated significantly with the sting reaction severity (r = 0.2752; P = .004) and with age (r = 0.2906; P = .002). Sting reaction severity also correlated with age (r = 0.3654; P = .001). CONCLUSIONS: Basal serum tryptase levels were found to be elevated in 11% of venom allergic patients and correlated significantly with both sting reaction severity and age. PMID- 17460944 TI - Effect of lysed Enterococcus faecalis FK-23 on allergen-induced immune responses and intestinal microflora in antibiotic-treated weaning mice. AB - BACKGROUND: Recent epidemiological studies have indicated that early life receipt of antibiotics may be associated with an increased risk of developing atopic disorder. Lysed Enterococcus faecalis FK-23 (LFK), a probiotic product of E faecalis, has been shown to have inhibitory effects on allergen-induced immune responses in mice. OBJECTIVE: The purpose of this study was to evaluate the effects of LFK on immune responses and intestinal microflora in antibiotic treated, and allergen-sensitized weaning mice. METHODS: Three-week-old BALB/c mice were sensitized with cedar pollen allergen to establish the experimental model. The allergen-induced peritoneal accumulation of eosinophils, serum levels of total and allergen-specific immunoglobulin (Ig) E and IgG2a, and the intestinal bacterial flora were determined in the control, antibiotic, LFK and antibiotic-LFK groups (n = 7 in all groups). Orally administered erythromycin, one kind of macrolide antibiotic, was used for the experiments. RESULTS: There was no significant difference in the allergen-induced peritoneal accumulation of eosinophils and serum specific IgE and IgG2a levels in erythromycin-treated mice compared to a control group. However, the ratio of serum total IgE to IgG2a levels was significantly increased in erythromycin-treated mice relative to that found either in LFK-treated mice or in erythromycin-treated mice with LFK supplementation. The total aerobes, total anaerobes and Enterococcus species of intestinal microflora were not significantly different among all groups. Lactobacillus species were distinctly eliminated in the mice exposed to erythromycin on day 7 and totally recovered in erythromycin-treated mice with LFK intervention on day 28, but could not be recovered in the erythromycin-treated mice without LFK intervention. CONCLUSIONS: Our results suggest that LFK may improve the intestinal ecosystem disturbed by antibiotic use, and thereby prevent subsequent development of atopy. However, whether different antibiotics have different effects on immune responses needs to be addressed further. PMID- 17460945 TI - Factors associated with allergic rhinitis in children from northern Mexico City. AB - BACKGROUND: The International Study of Asthma and Allergies in Childhood (ISAAC) questionnaire allows users to find factors associated with allergic diseases, but thus far most of the studies on risk factors for allergic diseases have been devoted to asthma and not to rhinitis. OBJECTIVE: To determine the main factors associated with symptoms of allergic rhinitis and rhinoconjunctivitis in school children and adolescents in northern Mexico City. PATIENTS AND METHODS: A cross sectional, multicenter survey was conducted in northern Mexico City, in children aged 6-7 and 13-14 years. The survey instrument was the Phase Three B ISAAC questionnaire, which was validated and standardized in Spanish. RESULTS: There were 4106 6-7-year-olds and 6576 13-14-year-olds. The total prevalence of diagnosis of allergic rhinitis was 4.6%. The prevalence of cumulative and current symptoms of rhinitis was considered high (>29%), but the prevalence of the diagnosis of allergic rhinitis was considered low (ranging from 3.4% to 5.6%). The prevalence of symptoms of rhinitis with conjunctivitis had intermediate values (ranging from 20.3% to 30.2%). Cumulative symptoms of allergic rhinitis, current symptoms of allergic rhinitis, and rhinoconjunctivitis were related to symptoms of current or cumulative asthma, symptoms of current or cumulative atopic eczema, and current use of paracetamol (odds ratio > 1, P < .05). CONCLUSION: The present results support the concept of rhinitis and asthma as common chronic respiratory diseases, and this study also found a relation between paracetamol use and rhinitis in children. PMID- 17460946 TI - Prevention of new sensitizations by specific immunotherapy in children with rhinitis and/or asthma monosensitized to house dust mite. AB - BACKGROUND: Previous studies have suggested that single-allergen-specific immunotherapy (SIT) may prevent sensitization to other airborne allergens in monosensitized children. We aimed to assess the prevention of new sensitizations in monosensitized children treated with single-allergen SIT injections in comparison with monosensitized patients given appropriate pharmacologic treatment for their disease. METHODS: A total of 147 children with rhinitis and/or asthma monosensitized to house dust mite were studied; 45 patients underwent SIT with adsorbed extracts and 40 patients underwent SIT with aqueous extracts for 5 years. The control group was comprised of 62 patients given only pharmacologic treatment for at least 5 years. Skin prick tests, medication scores for rhinitis and asthma, and atopy scores according to skin prick tests were evaluated at the beginning and after 5 years of treatment. RESULTS: All groups were comparable in terms of age, sex, and disease characteristics. At the end of 5 years, 64 out of 85 (75.3%) in the SIT group showed no new sensitization, compared to 29 out of 62 children (46.7%) in the control group (P = .002). There were no differences between the SIT subgroups with regard to onset of new sensitization (P = .605). The patients developing new sensitizations had higher atopy scores (P = .002) and medication scores for both rhinitis (P = .008) and asthma (P = .013) in comparison to patients not developing new sensitizations after 5 years of SIT. CONCLUSION: According to our data, SIT has the potential to prevent the onset of new sensitizations in children with rhinitis and/or asthma monosensitized to house dust mite. PMID- 17460948 TI - Tumor necrosis factor-alpha upregulates the expression of immunoglobulin secretory component. AB - BACKGROUND: The immunoglobulin (Ig) secretory component (SC) is the extracellular component of the polymeric Ig receptor (plgR) that is responsible for the transcytosis of newly synthesized IgA. In addition, the SC seems to play important roles in regulating eosinophil functions and in enhancing local immune responses. SC expression in HT-29 has been shown to increase in response to interferon-gamma, interleukin (IL) 4 and IL-1, but whether tumor necrosis factor (TNF) alpha affects SC expression is disputed. OBJECTIVE: Our aim was to study whether TNF-alpha can affect the expression of SC in Caco-2 cells. METHODS: We used immunocytochemistry, enzyme-linked immunosorbent assay, Western blot, and quantitative real-time polymerase chain reaction to test SC-positive cells, free SC in culture supernatants, plgR mRNA, and protein expression of SC. RESULTS: TNF alpha dose-dependently increased SC-positive cells, free SC in culture supernatants, plgR mRNA, and protein expression of SC. PMID- 17460947 TI - Fluctuation of fecal microbiota in individuals with Japanese cedar pollinosis during the pollen season and influence of probiotic intake. AB - BACKGROUND: We have previously reported the results of a randomized, double blind, placebo-controlled trial that found the intake of yogurt supplemented with a probiotic strain, Bifidobacterium longum BB536, alleviates symptoms and affects blood parameters in individuals with Japanese cedar pollinosis (JCPsis) during the pollen season. OBJECTIVE: In the present study, fecal microbiota were investigated to examine whether any changes occur during the pollen season and whether any influence is exerted by probiotic intake. METHODS: Yogurt either with BB536 (BB536 yogurt) or without BB536 (placebo yogurt) was administered for 14 weeks at 2 x 100 g per day to 40 subjects (17 men, 23 women) with a clinical history of JCPsis. Fecal samples were obtained from 23 subjects (placebo group, n=13; BB536 group, n=10) before and during the intervention (weeks 4, 9 and 13) and fecal microbiota were analyzed using terminal-restriction fragment length polymorphism and real-time polymerase chain reaction (PCR) methods. RESULTS: From the fluctuation patterns of terminal-restriction fragments, the Bacteroides fragilis group and bifidobacteria were among the species that changed most with pollen dispersion. Real-time PCR analyses indicated that the cell numbers of the B fragilis group increased significantly along with pollen dispersion in both BB536 and placebo groups. Cell numbers of bifidobacteria were significantly higher in the BB536 group compared with the placebo group (P < .05 at weeks 4 and 9). The ratio of cell numbers of the B fragilis group to bifidobacteria increased significantly during the pollen season in the placebo group (P < .01 at weeks 9 and 14), but not in the BB536 group. An in vitro study using peripheral blood mononuclear cells from JCPsis subjects indicated that strains of the B fragilis group induced significantly more helper T cell (T(H)) type2 cytokines (interleukin [IL]-6) but fewer T(H)1 cytokines (IL-12 and interferon) compared with those of bifidobacteria. CONCLUSIONS: These results suggest a relationship between fluctuation in intestinal microbiota and pollinosis allergy. Furthermore, intake of BB536 yogurt appears to exert positive ihfluences on the formation of anti-allergic microbiota. PMID- 17460949 TI - Apoptosis and asthma in the elderly. AB - BACKGROUND: Asthma is a chronic inflammatory disorder of the airways. The persistence of airway inflammation depends on a decrease in apoptosis of T lymphocytes and eosinophils and survival of these activated cells. T lymphocytes expressing gamma delta receptors can be identified in human lungs and play an important role in immune defence against pathogens and in the regulation of chronic inflammation. Aging is associated with evidence of some immune dysregulation. OBJECTIVE: The aim of this study was to analyze the apoptosis receptors of T lymphocytes in long-lasting asthma, to establish their correlation with activation markers such as CD25+ and human leukocyte antigen (HLA)-DR+, and to analyze the gama delta T cell expression in this disease. METHODS: A group of 64 individuals (group A) who had had asthma for more than 30 years (mean age [+/ SD] 72 +/- 5 years) and 61 healthy individuals acting as controls--group B with 41 individuals (mean age 79 +/- 7 years) and group C with 20 individuals (mean age 38 +/- 12 years) were included in the study. All subjects underwent clinical evaluation and spirometric testing. Peripheral blood cells were stained with monoclonal antibodies anti-CD3, anti-CD4, anti-CD8, anti-CD25, anti-TCR gamma delta, anti-HLA-DR and anti-CD95. Statistical comparisons were performed between the asthmatics and the elderly control group and between the elderly control group and the adult control group. RESULTS: The average percentage of predicted forced expiratory volume in the first second was 73.6 gamma delta 25.3. The mean values of T cell receptors for asthma group A vs elderly control group B vs adult control group C respectively, were the following: CD3, 74.9+/-7 vs. 74.8 +/- 8.8 (P=ns) vs. 76.7 +/- 4.2 (P=ns); CD4, 48.8 +/- 8.7 vs. 43.5 +/- 10.2 (P=ns) vs. 44.8 +/- 3.8 (P=ns); CD8, 23.3 +/- 7.9 vs. 25.7 +/- 10.2 (P=ns) vs. 25.6 +/- 4.5 (P=ns); CD25, 14.3 +/- 5.9 vs. 22.4 +/- 7.8 (P = .0001) vs. 5.5 +/- 2.4 (P = .0001); TCR gamma delta, 2.8 +/- 2.1 vs. 4.1 +/- 3.3 (P < .05) vs. 4.6 +/- 2.1 (P=ns); HLA-DR, 18.4 +/- 9.2 vs. 17.8 +/- 5.9 (P=ns) vs. 15.4 +/- 5.1 (P=ns) and CD95, 49.3 +/- 13.7 vs. 52.6 +/- 12.1 (P=ns) vs. 13.8 +/- 10.8 (P = .0001). CONCLUSIONS: The immunological and inflammatory changes related to ageing may cause an increase in CD95 and CD25 T cell expression. In asthma, blood cells may express increased activation and apoptosis markers but in elderly patients taking steroids, these receptors remain within normal ranges. The number of gamma delta T cells may be lower in long-lasting asthma, and have a limited modulatory effect on allergic inflammatory reactions. The evaluation of patients with long-lasting asthma should take into account the immunological and inflammatory changes present in the elderly in order to avoid results being misinterpreted. PMID- 17460950 TI - Prawns, barnacles, and nonsteroidal anti-inflammatory drugs: effect modifiers or diagnostic confounders [corrected]. AB - A 42-year-old woman with no history of atopy reported several episodes of generalized urticaria and shortness of breath after eating shellfish (prawns and barnacles) but with good tolerance of the same foods between episodes. Skin prick tests (SPTs), serum enzyme allergosorbent tests (EAST) for specific immunoglobulin (Ig) E, Western blot and inhibition assays, and oral challenge tests with prawns, barnacles, nonsteroidal anti-inflammatory drugs (NSAIDs), and alcohol as potential effect modifiers were performed. Specific IgE to both barnacle and prawn were detected by SPTs and EAST. Results from a Western blot of raw prawn revealed an IgE binding band of 37 kDa and IgE binding bands of 143, 83, 38, 32, and 20 kDa appeared in the raw barnacle assay. Oral challenge tests were positive with prawns and prawn extract only if preceded by NSAIDs. Oral challenges with NSAIDs alone, prawns alone, barnacles with or without NSAIDs and alcohol led to no reaction. A synergistic effect of NSAIDs in inducing anaphylaxis after prawn intake was confirmed. No similar effect was achieved with barnacles despite the presence of specific IgE. Additional factors needed to elicit a clinical reaction in food allergy may not be obvious and several oral challenge protocols are mandatory in such cases. PMID- 17460951 TI - Selective sensitization to clavulanic acid and penicillin V. AB - Allergic reactions to beta-lactam antibiotics have been reported frequently and may occur because of sensitization to unique haptens or to determinants shared with other drugs. A woman who received 1 tablet of amoxicillin-clavulanic acid developed wheals and flares although she had previously tolerated the same preparation well. Levels of specific immunoglobulin (Ig) E to penicillin V, penicillin G, amoxicillin, and ampicillin were undetectable. Skin tests to amoxicillin, penicillin major determinant and minor determinant mixture were negative. The patient tolerated oral challenge with 500 mg of amoxicillin but developed wheals and flares when challenged with amoxicillin-clavulanic acid 500/125 mg. A histamine release test was negative with amoxicillin but positive with the amoxicillin-clavulanic acid and clavulanic acid. A prick test to the combination was positive. Specific IgE to penicillin V later became positive while remaining negative to other beta-lactams. No inhibition was obtained using penicillin V against clavulanic acid and amoxicillin but was complete when penicillin V was used in the solid-phase and as the inhibitor. No cross reactivity was proven between these sensitizations. PMID- 17460952 TI - Detection of serum histamine-releasing factors in a patient with idiopathic anaphylaxis and multiple drug allergy syndrome. AB - We describe the case of a 30-year-old woman who reported several episodes of anaphylaxis with angioedema and relapsing urticaria. Some events were related to nonsteroidal anti-inflammatory drug intake and one episode followed alcohol ingestion, but in most cases no triggers could be identified. Specific immunoglobulin E determination was negative for food and drug allergens, C3 and C4 were in the normal range, C1 inhibitor was slightly reduced and serum tryptase was undetectable. In vivo autologous serum skin test and in vitro basophil histamine release assay were positive indicating the presence of circulating histamine-releasing factors. When oral tolerance tests were performed, only doxycycline was tolerated whereas levofloxacin, clarithromycin, nimesulide and tramadol caused mild urticaria. Premedication with cetirizine allowed the patient to tolerate levofloxacin, clarithromycin and nimesulide. The demonstration of circulating histamine-releasing factors in a patient with idiopathic anaphylaxis and multiple drug allergy syndrome provides a new mechanistic insight and might open the way to new therapeutic approaches. PMID- 17460953 TI - Hypersensitivity pneumonitis caused by domestic exposure to molds. PMID- 17460954 TI - Erytema multiform due to hyaluronic acid (go-on). PMID- 17460955 TI - Allergy to chicken in patients sensitized to Anisakis species. PMID- 17460956 TI - Codman Award 2006: the experience of hope in ALS patients. AB - In this study, the concept of hope in patients diagnosed with amyotrophic lateral sclerosis was explored. The researchers asked, "What does hope mean to you?" The final sample consisted of 13 patients at varying stages of illness and at differing times since diagnosis. Semi-structured interviews with open-ended questions were used to elicit views on hope, its sources and evolution. The findings were similar to a study done by Herth (1990) in which two themes emerged: hope fostering strategies and hope hindering barriers. The study process allowed the researchers to capture the participants' sources of hope. An unexpected finding was that participants described taking part in the interviews as "therapeutic." The findings support the notion that clinicians need to engage patients in discussions of hope, particularly the question of hope and its meaning, in order to adequately address their most important concerns and understand their point of view. Implications and suggestions for nursing practice include taking the time to explore hope-fostering approaches and discuss hope hindering obstacles and our role in creating a hopeful environment. PMID- 17460957 TI - Quality of life: Gamma Knife surgery and whole brain radiation therapy. AB - Cerebral metastasis of cancers originating outside the brain has traditionally been treated with whole brain radiation therapy (WBRT). Gamma Knife Radiosurgery (GKS) provides safe and effective alternative treatment that is less invasive and has fewer side effects. Both WBRT and GKS are reviewed and discussed in terms of quality of life and health outcomes. The case studies of two individuals who underwent Gamma Knife surgery are presented. PMID- 17460958 TI - Case study: closure of a patent foramen ovale without surgery following stroke. PMID- 17460959 TI - [Gluten-sensitive coeliac disease and prevention of autoimmune diseases and tumors]. PMID- 17460960 TI - [Epidemiology of gastroduodenal ulcer]. PMID- 17460961 TI - [Prevalence of Helicobacter pylori CaGa strains and peptic ulcer in the population of Eastern Siberia]. AB - AIM: To study prevalence of Helicobacter pylori (Hp) CagA strains and ulcer disease in native population of eastern Siberia and migrants. MATERIAL AND METHODS: Epidemiological screening of Hp CagA strains and ulcer disease was performed by cross-sectional method in population of Evenkia, Khakassia, Tuva. Fibrogastroduodenoscopy was made in 3602 examinees; IgG of CagA Hp was detected in the sera of 2472 patients with entyme immunoassay. RESULTS: Prevalence of ulcer was higher in Europeans than in Mongoloids. Detectability of CagA Hp was 36.5% among khakas population, 43.8% among evenks, 60.1% among tyvin population, 59.8% among European population. In khakas and tyva population with ulcer CagA Hp incidence was higher than in healthy subjects. CONCLUSION. CagA HP is associated with ulcer in mongoloid population. This was not confirmed for European populatioin of the Eastern Siberia. PMID- 17460962 TI - [Immunological and pathogenetic aspects of imunofan administration in aged patients with duodenal ulcer]. AB - AIM: To specify immunological and pathogenetic aspects of imunophan use in aged patients with duodenal ulcer (DU). MATERIAL AND METHODS: Imunophan was given to 24 DU patients (mean age 62.0 +/- 1.5 years), in whom the size of ulcer varied from 0.6 to 2.1 cm. RESULTS: The patients on anti-ulcer therapy plus imunophan had a pain relief median 6.2 +/- 0.2 days (p < 0.001), control ones--11.8 +/- 0.1 days. The median of scarring duration in the test group was 16.2 +/- 0.2 days (p < 0.001), in the controls--23.8 +/- 0.3 days. In 3 (30.0%) cases the scars were rough. The count of T-lymphocytes in the study group increased from 53.1 +/- 0.6 to 65.1 +/- 0.2% (p < 0.001), of T-helpers/inductors--from 27.8 +/- 0.2 to 38.5 +/- 0.3% (p < 0.001), of cytotoxic T-lymphocytes--from 18.5 +/- 0.5 to 27.3 +/- 0.3 (p < 0.001), of B-lymphocytes--from 12.3 +/- 0.2 to 19.1 +/- 0.1% (p < 0.001). The therapy including imunophan reduced concentration of malonic dialdehide by 23.5%, trienic conjugates by 61.6%; raised the level of superoxide dismutase 1.6-fold, catalase 1.4-fold, glutathione reductase by 41.9% (p < 0.001). Neither immune status nor LPO-AOD changed significantly in patients on the basic therapy alone. CONCLUSION: The results obtained evidence for a positive action of imunophan on inflammation, immune status and antioxidant defense. Therefore, imunophan can be recommended as an adjuvant of basic anti-ulcer therapy in elderly and senile patients. PMID- 17460963 TI - [Age and eradication treatment of duodenal ulcer]. AB - AIM: To study efficacy of Helicobacter pylori (Hp) eradication (different schemes) depending on the age of patients with Hp-associated duodenal ulcer (DU). MATERIAL AND METHODS: A total of 182 patients with Hp-associated exacerbated DU (101 males and 81 females) participated in the trial. The patients were divided into three groups according to their age: group 1 (18-39 years)--71 patients, group 2 (40-60 years)--71 patients, group 3 (61-74 years)--40 patients. Before and 4-6 weeks after treatment the patients were examined according to the standard scheme and with esophagogastroduodenoscopy (EGDS) including biopsy from the body and pyloric portion of the stomach. Seventy five patients received omeprasol+clarithromycin +moxicilline (scheme 1), 37 patients - eprasol+clarithromycin+tinidasol (scheme 2), 40 - eprasol+clarithromycin+furasolidon (scheme 3), 30 -- eprasol+clarithromycin+vicram (scheme 4). RESULTS: Hp dissemination in the pyloric stomach was independent of the age, nonatrophic and atrophic gastritis occurred with equal frequency. Contamination of the gastric body in the aged patients was moderate and mild. Atrophic gastritis incidence increased with age. After treatment according to scheme 1, Hp eradication was achieved in all the patients of group 3, 90%--group 1, 87%--group 2; according to scheme 2--100% patients of group 3, 90%--group 2 and 87%--group 1; according to scheme 3--all patients of groups 2 and 3, 90%--group 1; according to scheme 4--100% cases from groups 1 and 87% in group 2. CONCLUSION: Intensity of Hp dissemination in gastric mucosa does not depend on the age; it changes with deterioration of gastric mucosa atrophy and regress of acid production. Hp eradication efficacy in the same schemes of treatment is higher in presenile and senile patients who, therefore, can be treated with less doses of antibiotics. PMID- 17460964 TI - [Colon diverticula: origin, prevalence, clinical manifestations]. AB - AIM: To study prevalence of colon diverticula (CD) and phenotypical features of patients with this anomaly. MATERIAL AND METHODS: At the first stage of the study we analysed colon x-ray evidence for 3 years to assess prevalence of CD. At the second stage we examined 59 patients with CD. Sixty patients with idiopathic constipation and 48 patients with irritable colon syndrome served control. RESULTS: We discovered the following independent factors associated with a high CD risk: history of CD in the relatives of the patients (OR 24.6; 95% CI 5.23 116.23; p = 0.0001) and age of the patient (OR 2.5; 95% CI 1.03-1.13; p = 0.0010). Age-dependent CD occurrence, similar anomalies in the relatives, other structural gastrointestinal anomalies in them, hyperaminociduria suggest that CD represent a metabolic involutionary process with hereditary predisposition. CD manifested clinically with abdominal pain and problems with colon passage. Abdominal pain related to defecation occurred with low pain threshold to extension. Abdominal pain risk in CD patients is significantly associated with intestinal infections (OR 4.56; 95% CI 1.09-18.97; chi-square = 4.34; p = 0.0372) and connective tissue dysplasia syndrome (OR 2.21; 95% CI 1.23-3.95; chi-square = 7.08; p = 0.0078). Independent risk factors of constipation in CD are non-morning defecation (OR 8.68; 95% CI 1.55-48.51; chi-square = 6.06; p = 0.0138), connective tissue dysplasia syndrome (OR 1.77; 95% CI 1.0-3.20; chisquare = 3.65; p = 0.0501), sedentary life style (OR 7.42; 94% CI 1.24 - 44.57; chi-square = 4.81; p = 0.0284). CONCLUSION: Incomplete CD reflect colon involution. Abdominal pain associated with defecation has much in common with irritable colon syndrome. PMID- 17460965 TI - [Chronic heart failure: structural and microbiological changes in the colon]. AB - AIM: To study microbiocenosis of the parietal layer of the colon and feces, concentrations of endotoxin and proinflammatory cytokines in patients with chronic heart failure (CHF) of different functional classes vs. healthy subjects of the same age. MATERIAL AND METHODS: The trial includes 37 patients with ischemic CHF and 13 healthy volunteers. The examination comprised 6-min walking test, echocardiographic evaluation of the left ventricular ejection fraction, clinical state by a special scale, assay for C-reactive protein, endotoxin, fecal seeding, colonoscopy with biopsy and seeding. RESULTS: Gram-negative flora in the colon and parietal layer occurred in high concentrations correlating with severity of CHF. The examinees with CHF of functional class III-IV had elevated levels of circulating endotoxin and serum C-reactive protein. PMID- 17460966 TI - [Polyvirus hepatitides (prevalence, diagnosis, characteristics of virusological process)]. AB - AIM: To ascertain prevalence of chronic hepatitides (CH) of polyvirus etiology and to characterize a course of such polyvirus CH. MATERIAL AND METHODS: A total of 15000 patients with chronic diseases of the liver (CDL) of different etiology were examined for markers of hepatotropic viruses, 312 patients with CH aged 18 59 years were examined clinically, biochemically, virusologically and morphologically. RESULTS: CH of polyvirus etiology are rather prevalent. A replicative form of HBV has a significant effect on the activity of the pathological process in mixed hepatitis. Incidence of chronic HBV and HCV infection rises with age. Viral hepatitis A in drug addicts runs an atypical course. CONCLUSION: Adequate epidemiological analysis, diagnosis and antiviral treatment, follow-up of CDL of mixed viral etiology, monitoring of drug addiction, healthy life style are able to reduce prevalence of mixed hepatitis. PMID- 17460967 TI - [Functional condition of the hepatobiliary system in patients with biliary dyskinesia]. AB - AIM: To study function of the liver, sphincter system and the gall bladder (GB) in patients with biliary dyskinesia (BD). MATERIAL AND METHODS: Thirty-four patients with BD have undergone clinical, fibrogastroduodenoscopic examinations, fractional duodenal intubation with examination of the bile, ultrasonography, dynamic scintigraphy of the hepatobiliary system. RESULTS: The examination of the patients has detected GB hypotonia (65%), GB hypertension (35%), Oddi's sphincter hypertention (28%), Oddi's sphincter hypotonia (44%), Lutkens' sphincter hypertonicity (19%), Lutkens' sphincter hypotonia (13%), impairment of intrahepatic biliary tracts (65%), lithogenic bile (100%), duodenogastric reflux (41%). CONCLUSION: Consecutive use of duodenal tubing, ultrasonography and hepatobiliscintigraphy provide assessment of defects in the function of the liver, biliary tract and sphincter system in patients with BD. PMID- 17460968 TI - [Impairment of humoral regulation of exocrine pancreatic function in chronic pancreatitis]. AB - AIM: To elicit the role of cholecistokinin (CCK), biogenic amines, bile acids (BA) in development of functional pancreatic insufficiency (PI) in chronic pancreatitis (CP). MATERIAL AND METHODS: Blood concentrations of CCK, serotonin and acetylcholin, fecal concentration of elastase (E-1), BA spectrum in the blood and duodenal content were studied in 46 CP patients (20 patients with alcoholic pancreatitis--AP and 26 patients with biliary pancreatitis--BP) and 15 healthy controls. RESULTS: In AP patients E-1 fell to 78.4 +/- 6.3 mcg/g (severe exocrine PI), while in BP patients E-1 was 170.0 +/- 28.9 mcg/g. CCK in AP and BP decreased to 0.33 +/- 0.03 and 0.45 +/- 0.03 ng/ml, respectively (control--1.60 +/- 0.02 ng/ml, respectively, p < 0.05). AP and BP patients had a rise in the absolute concentration and percentage of the total fraction of the taurodioxicholanic acids to 10.2 +/- 1.6 and 15.0 +/- 2.3%, respectively, (control 9.5 +/- 1.2%) in duodenal bile. The concentration of glycocholic acid fell to 24.1 +/- 1.6 and 23.7 +/- 3.7%, respectively, (control--36.4 +/- 2.4%, p < 0.05). AP patients had more significant decrease of taurocholic acid--to 4.5 +/ 0.7% (control--9.2 +/- 0.7%, p < 0.05). In the peripheral blood of AP patients there was an elevated basal level of serotonin and acetylcholine in the presence of low cholinesterase activity. After meal, acetylcholine concentration lowered in high secretion of serotonin. CONCLUSION: Depending on severity of destructive changes in the pancreas, AP and BP patients had different degree of exocrine insufficiency which may be secondary to the absence of acetylcholine rise in the blood after meal. Alterations in the composition of the conjugates of cholic and taurodioxicholanic BA lead to alterations of CCK blood concentration and, therefore, to changes in exocrine pancreatic secretion. Imbalance between serotonin and acetylcholine levels after meal evidences for defects in conventional regulatory interrelations. Decreased threshold of nociceptors activation in simultaneous enhancement of afferent nociceptive flows may entail pain syndrome in CP. PMID- 17460969 TI - [Ultrasonographic assessment of chronic pancreatitis severity. Pancreatic index]. AB - AIM: To introduce the pancreatic index assessing severity of chronic pancreatitis (CP). MATERIAL AND METHODS: Ultrasonography was made in 28 patients with clinically documented diagnosis of CP. A total of 6 groups of ultrasonographic signs served the basis for calculation of scores for a complex formalized semiquantitative criterion--the pancreatic index (PI). Ultrasonographic signs- the size of the head, calcinates, structure, maximal size of the ductus pancreaticus, pseudocysts, etc. were assessed in scores from 1 to 3. Severity of the disease was classified according to total PI. RESULTS: The following correlation was found between severity of CP and the PI: in mild CP--57%, in moderate CP--71%, in severe CP--66%. CONCLUSION: Ultrasonographic PI objectively evaluates severity of CP course. PMID- 17460970 TI - [Comparative trial of efficacy of trimethasidine MB and 3-(2,2,2 trimethylhydrasine) propionate dihydrate in chronic heart failure]. AB - AIM: To study efficacy of the myocardial cytoprotector trimethasidine MB and metabolic drug 3-(2,2,2-trimethylhydrasine) propionate dihydrate (3-TMHP) in the treatment of chronic cardiac failure (CCF). MATERIAL AND METHODS: Sixty-five patients with CCF after myocardial infarction (> 6 months) with left ventricular ejection fraction (LV EF) <40% were randomized into 3 groups: group 1 (n=28) received basic therapy plus trimethasidine in a daily dose 70 mg; group 2 (n=25)- basic therapy plus 3-TMHP in a daily dose 1000 mg; control group (n=12) received basic therapy with ACE inhibitors, beta-blockers and diuretics. Before and after 6-month treatment all the patients have undergone stress echocardiography with dobutamine. Perfusion and myocardial metabolism were determined in 34 patients with single photon emission computed tomography of the myocardium (SPECT) with 99m-Tc-technetril and positron-emission tomography of the myocardium (PET) with F 18-fluorodesoxyglucose. RESULTS: Groups 1 and 2 significantly reduced functional class of CCF and prolonged the distance of a 6-min walk. Significant improvement of life quality was observed only in the treatment with trimethasidine. According to PET, treatment with trimethasidine MB and 3-TMHP has an anti-ischemic action manifesting with a significant attenuation of glucose hypermetabolism in the ischemic segment to normal values. However, significant improvement of systolic thickening in hybernated segments by SPECT as well as a significant rise of LV EF were recorded only in the treatment with trimethasidine MB. Stress echocardiography with dobutamine had high specificity (85.7%) but low sensitivity (50.4%) in detection of hybernated myocardium. CONCLUSION: Trimethasidine MB (preductal MB) has advantages over 3-TMHP, so it is preferable in ischemic CHF. PMID- 17460971 TI - [Pharmacological efficacy of bismuth tripotassium dicitrate in peptic ulcer]. PMID- 17460972 TI - [Replacement enzyme therapy in failure of exocrine pancreatic function]. PMID- 17460973 TI - [Multiorganic hemodynamic disorders in hepatic cirrhosis]. PMID- 17460974 TI - [Flat adenomas of the colon]. PMID- 17460975 TI - [Gastropathy caused by nonsteriod anti-inflammatory drugs: from understanding of development mechanisms to planning strategy of treatment and prophylaxis]. PMID- 17460976 TI - [The VI congress of Russian Gastroenterology Society]. PMID- 17460977 TI - [Falk Symposium in Moscow]. PMID- 17460978 TI - [Effect of volatile inhalational anesthetics on cerebral blood volume and oxygen status in children]. AB - The investigation evaluated the effect of various volatile anesthetics on cerebral blood volume and oxygen status in sick children at the stage of anesthesia induction. Ninety-two children were distributed into 3 groups: Groups 1 (n = 36) and 2 (n = 24) underwent stepwise induction with halothane and enflurane, respectively. Group 3 (n = 32) had vital capacity rapid inhalation induction with sevoflurane. Cerebral oximetry (NIRS method) was used to measure the content of hydroxyhemoglobin, deoxyhemoglobin, the total level of hemoglobin and to assess regional cerebral tissue saturation (rSO2). Halothane was ascertained to increase cerebral blood volume by 20.5% whereas enflurane and sevoflurane increased it only by 8.8 and 9.0%, respectively. In all cases, the value of rSO2 remained comparatively high, by exceeding the baseline level by 3 5%. PMID- 17460979 TI - [Modern inhalational anesthesia in pediatric maxillofacial surgery]. AB - The paper deals with the use of sevoflurane and isoflurane in elective pediatric maxillofacial surgery. Having outlined the major features of anesthesia in this category of patients, the authors emphasize the application of the conception of the minimum alveolar concentration (MAC) and its derivatives (MAC bar and MAC awake) reflecting the analgesic and hypnotic power of an inhalational anesthetic, respectively, and theoretically substantiate the use of certain anesthetic concentrations at particular stages of surgery. The technique described in the paper was used in 82 patients aged 1.5 months to 18 years, undergoing cleft lip and cleft palate repair, residual lip and nose deformation repair, and other elective maxillofacial procedures. The interventions lasted 40 minutes to 8 hours. Sevoflurane was commonly used as an induction agent while isoflurane was applied to maintain anesthesia. The anesthetics were administered at 1 to 1.3 and 0.8 MAC at traumatic and low traumatic stages of surgery, respectively. During the first traumatic period, the dose of fentanyl was not greater than 1.5 and 2.5 microg/kg in old and young children, respectively. Hemodynamics was characterized by moderate controlled hypotension with an average decrease in mean blood pressure by 30% of the age-related value. Emergence occurred 5-15 min after an anesthetic was discontinued. By and large, the technique demonstrated its efficiency and safety in this group of patients. PMID- 17460980 TI - [Use of the inhalational anesthetics sevorane and isoflurane in children]. PMID- 17460981 TI - [Assessment of a hypnotic component on the basis of BIS monitoring during balanced anesthesia in children]. AB - The hypnotic components of balanced anesthesia under regional anesthesia were estimated in 60 children during operations on the limbs and urogenital area. BIS monitoring was to assess the grade of sedation. Analysis of the findings showed that balanced anesthesia could ensure an adequate and stable anesthesia throughout the surgical intervention. The optimal BIS index for moderate sedation was 40-60 units. Among all the types of balanced anesthesia, sevoflurane-based anesthesia was most effective. PMID- 17460982 TI - [Balanced spinal-epidural anesthesia in pediatric orthopedics]. AB - The problem of spinal epidural anesthesia used in pediatric surgery has been inadequately studied. This investigation was undertaken to develop a balanced spinal epidural anesthesia schedule and to analyze its efficiency during orthopedic operations on the lower extremities in 119 children aged 2 to 16 years. Preference is given to sequential subarachnoidal and epidural space punctures via the same access. Comparative assessment of monitored indices during surgery and a need for postoperative analgesia in the groups of patients with conventional general anesthesia (n = 60) and those who had undergone spinal epidural anesthesia (n = 59) demonstrated a reliable intraoperative and early effective postoperative analgesia after balanced spinal epidural anesthesia. PMID- 17460983 TI - [Modern regional anesthesia during surgeries on the limbs in children]. PMID- 17460984 TI - [Anesthesia recovery period after anesthesia with ketamine and propofol during minor surgery in children]. AB - Forty-eight children aged 4 to 15 years, with an ASA I-II physical status, were examined. The patients underwent short mini-invasive surgical interventions under intravenous anesthesia using ketamine and propofol. The examinees were divided into 2 age groups: 1) 20 children aged 4 to 7 years; 2) 28 aged 8 to 15 years. The clinical picture of the postoperative period, central hemodynamics, acid-base balance, and blood gas composition were studied. The changes in the parameters were identical in both groups. The specific features of the course of emergence from propofol-ketamine anesthesia require that the patients should be postoperatively observed. PMID- 17460985 TI - [Use of the non-depolarizing myorelaxant vecuronium bromide (norkuron) in children]. PMID- 17460986 TI - [Central and cerebral hemodynamics during gynecological laparoscopic interventions in children]. AB - The authors examined central and cerebral hemodynamics in children during gynecological laparoscopic interventions. The course of 64 anesthesias was analyzed in girls aged 3 to 16 years, who had undergone laparoscopic surgery. Central hemodynamics was studied by tetrapolar rheography on a Diamant apparatus; cerebral oximetry was examined on a Critikon RedOx Monitor 2020 device. The findings demonstrate that pneumoperitoneum application causes immediate cardiovascular changes. Transfer of patients to Trendelenburg's position had a beneficial impact on central hemodynamics, by facilitating the normalization of venous return, thus maintaining cardiac output. Central hemodynamic disorders promptly affected cerebral blood volume and oxygen status. After pneumoperitoneum application, there was an average of 3% reduction in regional cerebral tissue saturation. PMID- 17460987 TI - [Causes of the critical status of neonatal infants with developmental malformations in intensive care units]. AB - There are many contradictory data available in the literature on the trends of diagnosed congenital malformations, including those requiring emergency treatment in neonatality. An examining specialist must make use of the algorithm to determine the condition of a neonate, which allows him to qualitatively define the time of surgical treatment and to predict the further course of disease, including its postoperative course. Problems can be solved in the context of evidence-based medicine. To compare the degree and causes of the critical condition of neonatal infants with developmental malformations, the authors have developed a severity rating scale for the newborn with developmental malformations at a gestational age of 35 weeks and a body weight of more than 2 kg, treated at an intensive care unit, by using the results of clinical, laboratory, and morphological studies and estimating the scores. Infants with esophageal atresia, diaphragmatic hernia, anterior abdominal wall and intestine were used as an example to determine changes and causes of the severity of neonates before and after surgery, by taking into account their scores. PMID- 17460988 TI - [Tracheal intubation in neonatal infants]. PMID- 17460989 TI - [Nutritive provision of children during intensive therapy in the early post aggressive period]. AB - The purpose of the study conducted at the intensive care units (ICI) of a Tushino children's city hospital (CCH), Moscow, in 2000-2005, was to enhance the efficiency of treatment in children with brain injuries, severe pneumonias, or appendicitis-induced peritonitis in the early postaggression period, by optimizing their feeding. Examination of 224 patients aged 1 month to 15 years, treated at the ICI of the Tushino CCH for brain injuries, severe pneumonias, or appendicitis-induced peritonitis in 2000-2005, indicated that mixed (parenteral and enteral) feeding was more effective in children in the early postaggression period than enteral feeding. Consideration of the size of protein losses and the amount of dietary protein and energy and estimation of nitrogen balance revealed that, by increasing the amount of dietary protein and energy, lowering protein losses, and thus producing positive changes in nitrogen balance, higher blood glucose decrease rates, and in a number of anthropometric and somatometric indices, mixed (parenteral and enteral) feeding is an effective method of nutritive provision in children at an intensive care unit. PMID- 17460990 TI - [Comparative evaluation of the efficiency of enteral and mixed (parenteral enteral) feeding in children with pneumonia in the intensive care unit]. PMID- 17460991 TI - [Free radical oxidation in children with severe craniocerebral injury]. AB - Changes in the products of lipid peroxidation (LPO) and the antioxidative system (AOS) were studied in 94 children aged 3 to 15 years with severe brain injury (BI) on days 1, 3-5, and 7-10. The contents of dienic conjugates and malonic dialdehyde and the level of induced peroxide hemolysis and peroxide resistance of red blood cells in the plasma were determined. Children with severe BI were found to have activated LPO processes and decreased AOS activity, the most significant changes occurring within 24 hours of injury. The increased activity of peroxide processes was attended by reverse AOS changes. In children, the time course of changes in the parameters of LPO and AOS differs from that in adults since there is no phase of excessive LPO activation due to the exhaustion of AOS reserves. PMID- 17460993 TI - [Pathomorphological pulmonary changes in patients with severe craniocerebral injuries]. PMID- 17460992 TI - [Endocrine homeostasis and its correction with stress protectors in children with severe burn injury]. AB - The use of stress protection with dalargin, clofelin, and pentamine in the treatment of children with severe burn injury was ascertained to result in a reduction in the magnitude of abnormal changes in the endocrine system and in the prevention of depletion of its compensatory capacities. This treatment will assist in reducing the severity of endotoxicosis, as well as the frequency and severity of toxic and pyoseptic complications. PMID- 17460994 TI - [Algorithm of therapy in neonates with developmental malformations depending on causes and severity of illness in the intensive care unit]. AB - The results of treatment for congenital malformations requiring emergency interventions depend on the timeliness and correctness of diagnosis of a defect, on the performance of resuscitative and surgical interventions, the correct evaluation of a neonate's condition at different stages of treatment. The score rating of the patient's condition is objective and may be an evidence-based medicine tool. Prior to the emergence of the universal scale, many authors used their own developments. On the basis of achievements made by pediatric surgeons, anesthesiologists, resuscitators in the context of statistical analysis, the specific features of neonatology, microbiology, and clinical pharmacology, the authors of the paper have developed a neonatal severity rating scale with inclusion criteria, such as a body weight of more than 2 kg; intensive care unit (ICU) stay, artificial ventilation of at least 2 days of life. Life-threatening periods have been defined in neonatal infants treated at ICU; the main causes of their critical condition have been indicated in certain developmental malformations before and after surgery, the causes of death have been revealed from the material available in their case histories and from morphological charts. The purpose of the present study was to develop an algorithm of therapy for the newborn with severe developmental malformations at ICU and to define a promising line for increasing the survival rates in neonatal infants with these defects. PMID- 17460995 TI - [Prevention of pain during manipulations in neonatal infants: analysis of physicians' questionnaire survey]. AB - The paper presents the results of an anonymous questionnaire survey among the physicians of 7 children's health care facilities, who should assess and prevent pain in neonatal infants from 5 Russia's cities (Moscow, Cheboksary, Kursk, Novosibirsk, and Kyzyl). The questionnaire survey has indicated that all responding physicians agree with the statement that a neonate senses pain, but the severity of pain being uncontrolled and the pain syndrome unevaluated, pain analgesia and prevention are inadequately applied by comfort measures, particularly the use of glucose through a nipple, a pacifier, diapering, when procedures are performed. All the physicians are unanimous that analgesics or comfort measures should be more frequently used during any manipulations and procedures irrespective of whether they are performed for a long or short period. PMID- 17460996 TI - [Midazolam in children. Alternative routes of administration]. PMID- 17460997 TI - [Use of myorelaxants in children during neonatal period]. PMID- 17460998 TI - [High-frequency oscillatory lung ventilation in the treatment of adult patients]. PMID- 17460999 TI - Nutrition labels and claims in New Zealand and Australia: a review of use and understanding. AB - OBJECTIVE: To determine how well consumers use and understand nutrition labels and claims in New Zealand and Australia. METHOD: A review was undertaken of the literature on nutrition labelling in New Zealand and Australia published up to the end of July 2005. Major electronic databases were searched for appropriate literature and research, as were the bibliographies of relevant publications and pertinent websites. Studies that focused on health claims were excluded. RESULTS: Sixteen papers were suitable for inclusion in the review. All but one study evaluated self-reported use and understanding of nutrition labels. The single study that evaluated actual (observed) use of labels while shopping found frequency to be much lower than would be expected based on self-reported data. While self-reported understanding of nutrition labels was common, actual (evaluated) understanding appeared moderate at best. CONCLUSIONS: Self-reported use of nutrition labels and claims is common in New Zealand and Australia, but actual use and understanding appears limited. Nutrition labels present an opportunity to improve consumer food choice at point of purchase, but their potential value is limited by apparent lack of consumer understanding. IMPLICATIONS: Nutrition labels are an important part of a supportive environment that empowers people to make healthy food choices. Improving their ease of use and understanding has the potential to promote healthier food choices. PMID- 17461000 TI - An outbreak of chickenpox at a child care centre in Western Australia. Costs to the community and implications for vaccination policy. AB - OBJECTIVE: Between May and June 2002 an outbreak of chickenpox (CP) occurred at a child care centre in Perth, Western Australia. An epidemiological study was undertaken in order to determine the characteristics of the outbreak, assess vaccine effectiveness, and to define the direct and indirect costs associated with CP infections in young children. METHODS: A cohort study of the outbreak utilising attendance records and a telephone survey of parents was conducted. RESULTS: Of the 211 children attending the child care centre at the time of the outbreak, 44 contracted CP (attack rate 25.7%). In addition, two staff members, five secondary household contacts (secondary attack rate 38.5%) and four secondary non-household associated contacts were infected. There were no severe complications or any hospitalisations recorded in infected persons. Two cases had been vaccinated previously. Vaccine effectiveness for CP of any severity was 78.0% (95% CI 15.4-94.3%) while vaccine effectiveness against severe CP was 100%. Direct costs during this outbreak were estimated to be $54 per case and the total costs, including cost of parental time off work or study, were estimated to be $525.73 per case. CONCLUSIONS AND IMPLICATIONS: Although morbidity associated with CP in young children is not great, infection in childhood is almost universal. This study found that the average costs associated with each CP case were considerable. Since varicella vaccine affords good protection against CP, the recent inclusion of this vaccine in the Australian childhood vaccination schedule should save the community a considerable amount in direct and indirect costs if high coverage rates can be achieved. PMID- 17461001 TI - Patterns of psychostimulant prescribing to children with ADHD in Western Australia: variations in age, gender, medication type and dose prescribed. AB - OBJECTIVE: To examine treatment with psychostimulant medicines for attention deficit hyperactivity disorder (ADHD) in children (3-17 years) within Western Australia (WA) during 2004. METHODS: Using whole-population-based administrative pharmacy data for 2004, age-specific prevalence of children treated with dexamphetamine and methylphenidate was determined. Treatment with immediate action and sustained-release stimulants was also examined. Average daily dose (absolute and relative to body weight) of dexamphetamine, methylphenidate and total stimulant medication was also calculated. RESULTS: 9,658 children from 3-17 years (23.5 per 1,000 child population) were treated with stimulant medication for ADHD during 2004. Age-specific prevalence of treatment increased rapidly from three to eight years to peak at 14 years. The male-to-female ratio of stimulant treatment was 4.1. Child prevalence of treatment with dexamphetamine, methylphenidate or combined therapy was 16.4, 8.9 and 1.7 per 1,000 children, respectively. Most children were treated with immediate-action stimulants (16.9 per 1,000 child population), with fewer using sustained-release medicines (4.0 per 1,000 child population). CONCLUSIONS: The prevalence of stimulant medication use for the treatment of ADHD in WA children is 2.4%, with males accounting for the majority of treated patients. Most children were treated with dexamphetamine. Prescribing of methylphenidate, sustained-release stimulants or combined medication therapy was less common. Age-related trends in prescribed daily dose differed for dexamphetamine and methylphenidate. PMID- 17461002 TI - Evaluation of a prison-based hepatitis B immunisation pilot project. AB - This paper describes the evaluation findings of a hepatitis B immunisation pilot project, which aimed to increase the uptake and compliance of hepatitis B vaccinations among female prisoners in Victoria. The evaluation used a mixed methods approach including in-depth interviews, focus group discussions and an analysis of quantitative data. Fifty-five per cent of potential participants (391/712) were offered hepatitis B immunisation. Of those offered immunisation, 204 were eligible for immunisation and 169 (83%) received the first dose. Ninety three per cent of eligible women received two doses and 84% completed the three dose series. Lessons learnt from the pilot led to the revision of key prison hepatitis B immunisation policies and practices to ensure uniformity across Victorian prisons. PMID- 17461003 TI - Comparisons of trends in antiretroviral use and HIV notification rates between three Australian states. AB - OBJECTIVE: Notification rates for HIV among men who have sex with men (MSM) have been increasing, but this increase has not been uniform across States and Territories. This paper aims to determine whether differences in the levels of antiretroviral therapy (ART) were associated with differences in HIV notification rates between Australian States. METHODS: An ecological analysis of HIV notification and S100 drug prescribing data between 1998 and 2005. MAIN RESULTS: HIV notification rates in 2005 per 100 individuals living with HIV were lowest in New South Wales (NSW) (4.5) and higher in Victoria (6.9) and Queensland (7.8). The proportion of individuals receiving ART in 2005 was lowest in NSW (40%) and higher in Victoria (60%) and Queensland (60%). There has been a significant decline in the proportion of individuals living with HIV receiving effective ART per year in NSW (p < 0.01) and Queensland (p < 0.01), but an increase in Victoria (p < 0.01). CONCLUSION: Those States with the highest HIV notifications per 100 individuals living with HIV did not have the lowest proportion of individuals receiving ART. IMPLICATIONS: These data indicate that the differences in HIV notifications between States do not appear to be due to differences in ART. However, changes in ART over time in some States may be contributing to the rise in HIV notifications. PMID- 17461004 TI - Termination of pregnancy: associations with partner violence and other factors in a national cohort of young Australian women. AB - OBJECTIVE: To examine the associations between termination and other reproductive events, socio-demographic characteristics and experience of violence among a community-based national sample of young Australian women. METHODS: Using multiple logistic regression, we analysed data from the Younger cohort of the Australian Longitudinal Study on Women's Health comprising 14,776 young women aged 18-23 in survey 1 (1996), of whom 9,683 aged 22-27 also responded to survey 2 (2000). We stratified respondents into those aged below 20 and those who were older at survey 1. We compared the characteristics associated with terminations among teenage women in 1996 (survey 1) with those of women aged over 20 in 1996 who had not then reported a termination and who responded to survey 2 in 2000. Finally, we compared the characteristics of women reporting terminations, births, preterm births and miscarriages. RESULTS: Women reporting teenage terminations were more likely to be in a de facto relationship (OR = 1.94, 95% CI 1.17-3.21), less well educated (OR = 2.32, 95% CI 1.44-3.74), have no private health insurance, and be a victim of partner violence (OR = 3.11, 95% CI 1.76-5.49). Women reporting later terminations were also more likely to be abused by a partner (OR = 3.52, 95% CI 2.14-5.81).The relationship with violence held for the other reproductive events. CONCLUSION AND IMPLICATIONS: Partner violence is a strong predictor of termination and other reproductive outcomes among young Australian women. Education has a protective effect. Prevention and reduction of partner violence may reduce the rate of unwanted pregnancy. PMID- 17461005 TI - Partner violence prevalence among women attending a Maori health provider clinic. AB - OBJECTIVE: To determine partner violence rates among women attending a general practice in Aotearoa, New Zealand. METHODS: This descriptive study was conducted in a hauora (Maori health provider general practice clinic) in one South Auckland community. Non-acute, English-speaking women who entered the huaora during 30 randomly selected clinic sessions in a five-week period in 2003 were eligible to participate. Research assistants (RAs) verbally administered a structured, brief questionnaire that included a partner violence screen (past 12 months), assessment of high danger risk, and lifetime prevalence. Of 148 women approached, 109 participated. Participants generally self-identified as Maori (74%) or New Zealand European (18%) and ranged in age from 17 to 82 years (mean 38.8). RESULTS: Twenty-three per cent (95% CI 15-31) of women screened positive for partner violence. Among the 25 women who screened positive, six (24%) had one or more high danger risk factors and 24 (96%) reported one or more children living in the household. Seventy-eight per cent (95% CI 70-86) of women reported a history of partner violence. CONCLUSIONS: In this sample of mostly Maori women, direct partner violence questioning in a general practice setting yielded a high disclosure rate. Three out of four women disclosed violence by a partner; nearly one out of four disclosed violence by a partner in the past year. IMPLICATIONS: Healthcare providers have the opportunity to identify and provide services to women and their children experiencing partner violence. Health care providers and the health care system also have a responsibility to join with the community in calling for non-tolerance of family violence. PMID- 17461006 TI - Benchmarking epidemiological characteristics of cervical cancer in advance of change in screening practice and commencement of vaccination. AB - OBJECTIVES: To investigate trends in cervical cancer incidence, mortality and survival by histology for benchmarking purposes ahead of practice change and the introduction of Human Papilloma Virus (HPV) vaccine. METHODS: Using data from the South Australian Cancer Registry, age-standardised rates are presented for four year periods from 1977 to 2004. Socio-demographic and secular predictors of glandular as opposed to squamous cancers are investigated, using multivariable logistic regression. Disease-specific survivals are analysed using Kaplan-Meier product-limit estimates and Cox proportional hazards regression. RESULTS: Incidence and mortality rates reduced by 55.1% and 59.3% respectively between 1977-80 and 2001-04, with larger reductions for squamous than glandular cancers. The ratio of squamous to glandular cancer incidence reduced from 5.4:1 in 1977-88 to 2.8:1 in 1993-2004, with a corresponding reduction from 5.2:1 to 3.0:1 for mortality. Compared with squamous cancers, glandular lesions were more common in patients from higher socio-economic areas, but less common in those over 70 years of age, Aboriginal patients, and those born in Southern Europe. CONCLUSION: The proportion of cancers comprising glandular lesions has increased, possibly reflecting prevention of squamous cancers through treatment of screen-detected preinvasive lesions. Additional mortality reductions from screening may be limited where the proportion of glandular lesions is high, with vaccination offering the best prospects for gains in the long term. Priority should be given to Aboriginal and Torres Strait Islander women in vaccination programs in view of their high death rate from cervical cancer. PMID- 17461007 TI - Do healthy and unhealthy behaviours cluster in New Zealand? AB - OBJECTIVE: To describe the co-occurrence and clustering of healthy and unhealthy behaviours in New Zealand. METHOD: Data were sourced from the 2002/03 New Zealand Health Survey. Behaviours selected for analysis were tobacco use, quantity and pattern of alcohol consumption, level of physical activity, and intake of fruit and vegetables. Clustering was defined as co-prevalence of behaviours greater than that expected based on the laws of probability. Co-occurrence was examined using multiple logistic regression modelling, while clustering was examined in a stratified analysis using age and (where appropriate) ethnic standardisation for confounding control. RESULTS: Approximately 29% of adults enjoyed a healthy lifestyle characterised by non-use of tobacco, non- or safe use of alcohol, sufficient physical activity and adequate fruit and vegetable intake. This is only slightly greater than the prevalence expected if all four behaviours were independently distributed through the population i.e. little clustering of healthy behaviours was found. By contrast, 1.5% of adults exhibited all four unhealthy behaviours and 13% exhibited any combination of three of the four unhealthy behaviours. Unhealthy behaviours were more clustered than healthy behaviours, yet Maori exhibited less clustering of unhealthy behaviours than other ethnic groups and no deprivation gradient was seen in clustering. DISCUSSION: The relative lack of clustering of healthy behaviours supports single issue universal health promotion strategies at the population level. Our results also support targeted interventions at the clinical level for the 15% with 'unhealthy lifestyles'. Our finding of only limited clustering of unhealthy behaviours among Maori and no deprivation gradient suggests that clustering does not contribute to the greater burden of disease experienced by these groups. PMID- 17461008 TI - Australian smokers support stronger regulatory controls on tobacco: findings from the ITC Four-Country Survey. AB - OBJECTIVE: To examine Australian smokers' attitudes towards regulation of the tobacco industry and to compare their attitudes with those of three similar countries--the United Kingdom (UK), the United States (US), and Canada. METHOD: A telephone survey of 2,056 adult Australian smokers and 6,166 Canadian, US, and UK smokers was conducted in 2004 as the third wave of the International Tobacco Control Policy Evaluation Four-Country Survey. RESULTS: Australian smokers display the strongest support for regulation. Only 16% believe that tobacco companies should be allowed to advertise/promote cigarettes as they please, 70% agree that tobacco products should be more tightly regulated, and 64% agree that governments should do more to tackle the harms of smoking. Smokers see government failure to do so in cynical terms -77% agree that governments do not really care about smoking because of money from tobacco taxes. Opposition comes largely from smokers who hold self-exempting beliefs about smoking's risks, have a positive attitude to smoking, do not accept that smoking is socially denormalised, and do not hold tobacco companies responsible for harms caused by smoking. CONCLUSIONS AND IMPLICATIONS: The majority of Australian smokers believe that the tobacco industry is partly responsible for the predicament they find themselves in and want governments to act more strongly in their real interests. The strong relationship between support for regulation and cynicism about government inaction should stimulate governments into action. PMID- 17461009 TI - Annual health assessments for older Australian women: uptake and equity. AB - OBJECTIVE: To measure utilisation of Enhanced Primary Care (EPC) health assessment items for women aged 75 years and over, and to describe health and socio-demographic characteristics of users and non-users. METHOD: Analysis of longitudinal survey and Medicare claims data from women in the Australian Longitudinal Study on Women's Health (ALSWH) aged 75 to 78 years when EPC items were introduced and who provided permission to access their Medicare records for the period 1999-2003 (n = 4,646). RESULTS: There was an increase in uptake of assessments over four years: from November 1999, 12% of eligible women had a health assessment during the following year; by October 2003, 49% had at least one health assessment ever. Few had repeat assessments. Women who visited a GP more often and who were satisfied with the number of GPs available were more likely to have an assessment in the first 12 months, and women who visited a GP more often, those taking more medications, and those caring for another were more likely to have at least one assessment in four years. Women in smaller rural and remote areas were less likely to have an assessment than women in urban areas. CONCLUSIONS: Most women are not having annual assessments and there is some geographic inequity. PMID- 17461010 TI - Stressful life events, resources, and access: key considerations in quitting smoking at an Aboriginal Medical Service. AB - OBJECTIVE: Aboriginal and Torres Strait Islander people experience adverse health outcomes and have high rates of smoking and related illnesses. This brief report describes stress as a barrier to quitting smoking derived from reflections within an Aboriginal Medical Service and makes recommendations for intervention development. METHODS: A high-intensity smoking cessation program was conducted within a suburban Aboriginal Medical Service in Western Sydney, Australia, over a 10-month period. The intervention included weekly cessation counselling sessions and dispensation of free nicotine replacement therapy (NRT). RESULTS: During the observation period, 32 clients made quit attempts. To date, three clients (9%) have quit smoking. Chronic and intercurrent life stressors were noted to be the main barriers to smoking cessation described by participants. CONCLUSIONS: Achieving smoking cessation among Indigenous people is made significantly more complex because of multiple life stressors experienced. IMPLICATIONS: Future interventions targeting Indigenous Australians should take greater account of stressful life events and their impact on quitting smoking. PMID- 17461011 TI - Quantifying the excess risk for proteinuria, hypertension and diabetes in Australian Aborigines: comparison of profiles in three remote communities in the Northern Territory with those in the AusDiab study. AB - OBJECTIVE: To estimate the magnitude of excess risk for proteinuria, high blood pressure and diabetes in Australian Aboriginal adults in three remote communities by comparing them with nationwide Australian data. METHODS: Adult volunteers from three remote communities in the Northern Territory were screened for proteinuria, high blood pressure, and diabetes between 2000 and mid 2003. Rates for people age 25 to 74 years were compared with those from the AusDiab study conducted in 1999 and 2000. RESULTS: Compared with AusDiab, rates of these conditions were elevated in all Aboriginal communities, but differed among them. With adjustment for age and sex, rates of proteinuria were elevated 2.5- to 5.3-fold, rates of high blood pressure were elevated 3.1- to 8.1-fold and rates of diabetes were elevated 5.4- to 10-fold (p < 0.001 for all). The risk of having any condition ranged from 3.0- to 8.7-fold and the risk of having two or more conditions ranged from 5.8- to 14.2-fold. DISCUSSION: The data are compatible with the excess morbidity and mortality from cardiovascular disease, diabetes and renal disease in these Aboriginal groups. They reflect the multitude of risk factors operating in these environments. They dictate urgent and systematic intervention to modify outcomes of established disease and to prevent their development. However, the resources required for effective secondary intervention will differ among communities according to the disease burden. PMID- 17461012 TI - Northern Territory indigenous life expectancy improvements, 1967-2004. AB - OBJECTIVE: To assess the extent of changes in life expectancy at birth for the Indigenous population of the Northern Territory (NT) over the period 1967-2004, and to determine which age-specific mortality rates were mostly responsible for such change. METHODS: Life expectancy at birth figures were obtained via life table calculations using a high-quality and internally consistent dataset of NT Indigenous deaths and populations covering the period 1967-2004. A life expectancy at birth age decomposition technique was then applied. RESULTS: Indigenous life expectancy at birth has risen considerably in the NT, increasing from about 52 years for males and 54 years females in the late 1960s to about 60 years for males and 68 years for females in recent years. Significantly, for NT Indigenous females the gap with total Australian life expectancy has narrowed. CONCLUSIONS: In contrast to popular perception, Indigenous life expectancy in the Northern Territory has improved substantially from the late 1960s to the present. IMPLICATIONS: The widespread pessimism that surrounds Indigenous health and mortality is largely unfounded, at least for the NT. Although much remains to be done to reduce Indigenous mortality, the results in this paper demonstrate that improvements are occurring and that sustained and increased effort is worthwhile and will succeed. PMID- 17461013 TI - Fat chance for Mr Vegie TV ads. PMID- 17461014 TI - Too much has changed for old data to help today's OHS practitioner. PMID- 17461016 TI - [Effect of otoliths upon function of the semicircular canals after long-term stay under conditions of microgravitation]. AB - Results of studies performed in 12 astronauts after flights revealed that in some of them re-adaptation to the Earth gravitation involved increase of spontaneous oculomotor activity in immobile position of the head, suppression of the otolith function in static 40 degrees bending of the head towards right or left shoulders, enhancement of vestibular responsiveness in rotation of the head around the body longitudinal axis at the rate of 0.125 Hz. PMID- 17461017 TI - [Study of the letter and word recognition by the brain-computer-interface with P300 wave of human visual evoked potential]. AB - In four adult healthy subjects in 18 experiments, we studied Brain-Computer Interface recognition of different intended words by P300 wave in the VEP. The set of optimal characteristics of visual stimulation which rise reliability of recognition up to 100 %, as well as effective registration locus (Pz) were determined. It was found that the best processing criteria for letter recognition were: P300 square and superposition of all three criteria (P300 amplitude, square and covariation coefficient). PMID- 17461018 TI - [Parallel analysis of c-Fos protein and interleukin-2 expression in hypothalamic cells under different influence]. AB - The objective of this work was to perform a parallel analysis of activation of the rat anterior hypothalamus cells as judged by c-Fos protein expression, and of the expression of interleukin-2 (IL-2) under different influences, i. e., mild stress (handling) and adaptation to it, and intranasal administration of saline and the peptides Vilon (Lys-Glu) and Epithalon (Ala-Glu-Asp-Gly). Changes in the counts of cells positive for c-Fos- and IL-2 proteins were studied in structures of the lateral (LHA) area, anterior (AHN), supraoptic (SO) and paraventricular (PVH) nuclei of Wistar rat hypothalamus. Quantity of the interleukin-2-positive and c-Fos-positive cells was calculated. The findings were: a negative correlation between the activation of cells and the amount of IL-2 in the cells in the hypothalamic structures under study, and the specific patterns of changes in the counts of cells positive for c-Fos and IL-2 under stress and adaptation to stress. PMID- 17461019 TI - [Cerebral hemodynamic reaction to physical exercise of moderate intensity]. AB - The aim of this research was to study the cerebral hemodynamic reaction to stepped increase of physical exercises during the bicycle ergometer test in 12 young healthy male patients. The starting value of exercise was 0.25 W/kg of the body weight, with the 0.25 W/kg increase at every subsequent step up to the value of 1.75 W/kg of the body weight. Hemodynamic parameters were registered with the Doppler ultrasonography of middle cerebral artery before the study, during the last 10 seconds of every step, and during the 3 minutes of restorative period with a 1-minute interval. The peak systolic blood flow increase in the middle cerebral artery was observed only as the result of low intensity exercises (0.25 W/kg of the body weight). The blood pressure (BP) restoration occurred by the end of the third minute of the rest, while the cerebral hemodynamic indices became normal during the first minute. The research revealed a correlation between increases of vascular resistance caused by physical exercises and the BP, and no correlation between increases of peak systolic blood flow and BP, which displays the phenomenon of cerebral circulation autoregulation. PMID- 17461020 TI - [Cardiovascular peculiarities in hypertensive rats (Hsiah strain) after antihypertensive drug intervention during juvenile period of life]. AB - Treatment with different types of antihypertensive drugs during second month age has not prevented development of arterial hypertension and myocardial hypertrophy in the adult rats with inherited stress-induced arterial hypertension. At 6 months of age, the 11% attenuation of basal blood pressure has beet achieved only in the rats treated with angiotensin-converting enzyme inhibitor enalapril. Nevertheless, they expressed the most pronounced left ventricular hypertrophy. The unfavorable tissue and ultrastructural abnormalities were revealed in the myocardium of the rats which received the alpha1-adrenoceptor blocker terazosin. The delayed effects of losartan (angionetsin II receptor antagonist) and corinfar (calcium channel antagonist) on the myocardial structure were inessential. PMID- 17461021 TI - [Dependence of the circadian organization of heart rate regulation by prolonged musical sensory influence]. AB - It's shown that prolonged acoustic sensory influence in the form of classic music may change neurovegetative control of heart rate, enhancing the parasympathetic influences on heart rate variability, especially in night. These alterations increase expression of diurnal rhythm of ergotropic and trophotropic systems activity and lead to normalization of circadian profile of heart rate. PMID- 17461022 TI - [Role of cholinergic mechanisms of the ventrolateral preoptico-anterior hypothalamic area in regulation of sleep and wakefulness states in pigeons]. AB - Maintenance of wakefulness is established to accomplish muscarinic (M-) cholinergic receptor activation in the ventrolateral preoptic area of the hypothalamus. The "muscarinic" wakefulness is characterized by enhancement of electroencephalogram (EEG) power spectra in the 0.75-12 Hz band and by increase in brain temperature. Activation of nicotinic (N-) cholinergic receptors of the area produces an increase in the duration of slow wave sleep, EEG power spectra reduction in the 0.75-7 Hz band, a decrease in brain temperature. And its hyperactivation leads to wakefulness, during its episodes the brain temperature decreases. During M- and N-cholinergic receptor blockade, the sleep-wakefulness and thermoregulation changes opposite to their activation were found. It is suggested that M- and N-cholinergic receptors of the ventrolateral preoptic area in pigeons participate in the sleep-wakefulness regulation and this effect is related to influence of this area on GABA-ergic system. PMID- 17461023 TI - [Antinociceptive properties of the 5-HT3 receptor antagonist in the model of inflammatory pain in rats of different age with prenatal deficit of serotonine and under stress]. AB - The role of peripheral 5-HT3 receptors in the nociceptive behavioral response and the effect of the 5-HT3 antagonist ondansetron on indices of acute and tonic pain were investigated in the formalin test in 25- and 90-day-old Wistar male rats. The experimental rats were prenatally exposed to 5-HT depletion (a single injection ofparachlorophenilalanine 400 mg/kg/2 ml, i. p.; ICN, USA to the dams on day 9 of pregnancy) and to stress (dams immobilization during the last week of pregnancy). Antinociceptive effects of ondansetron in the rats with both prenatal 5-HT deficiency and stress (experimental rats) and prenatal injection of saline solution and stress (control rats) were more obvious in the younger animals. Prenatal 5-HT deficiency attenuated the antinociceptive effect of ondansetron in licking patterns in the younder age group in acute pain, and in adults--in tonic pain. Thus, the data obtained in the rats with prenatal 5-HT deficiency and stress indicate involvement of 5-HT3 receptors in mediation of prolonged pain in the formalin test, and antinociceptive effect of ondansetron which is attenuated in animals with prenatal 5-HT deficiency and specifically depends on rat's age. PMID- 17461024 TI - [V. N. Chernigovsky and organisation of physiological investigations in the USSR]. PMID- 17461025 TI - [The essay on history of studying the renal functions and the organisation of these studies in the USSR and in Russia]. PMID- 17461026 TI - [Dietary fatty acid intake and concentrations in serum and erythrocyte membranes of students with allergic disease]. AB - PURPOSE: An attempt was made to utilize accumulated scientific evidence for nutritional guidance concerning dietary fatty acids (FAs) for students with allergic diseases METHODS: A questionnaire survey on dietary fatty acid intake was conducted with 128 women students aged 19-20. In addition to hematological and physical examinations, fatty acid analyses were performed with serum and erythrocyte membrane samples using gas chromatography. Based on the answers to questions about allergic diseases, the subjects were divided into three groups (59 healthy students, 45 with previous experience of allergies, and 24 with allergies). We then investigated the influences of dietary fatty acids and other nutritional components on fatty acid compositions of serum and erythrocyte membranes, and statistically analyzed the results by comparing the three groups. RESULTS: 1) Dietary n-3 (g) intake by all students was lower than the tentative dietary goal in the Dietary Reference Intakes. However, there were no effects of allergic diseases on physical measurements and blood test data. Dietary n-3 (g) was negatively correlated with erythrocyte membrane n-3 (%), and dietary n-3 (%) was positively correlated with the number of acidophils (%). In addition, a positive correlation was found between serum n-3 (%) and dietary S (Saturated fatty acid) (%). 2) For the allergic group, the ratio of erythrocyte membrane M (Monounsaturated fatty asid) (%) to dietary M (%) was high. For the allergic predisposition group (allergic subjects and subjects with past history of allergic disease), a negative correlation was found between erythrocyte membrane M (%) and dietary S (%). CONCLUSION: Dietary n-3 (g) was insufficient in all subjects enrolled in this study, but erythrocyte membrane n-3 (Y%) decreased with increase of dietary n-3 (g). There was a tendency for acidophil number (%) to increase with the dietary n-3 (%). Therefore, it was suggested that if nutritional guidance is to made with attention to increasing dietary S (%), it should be stressed that serum n-3 (%) might become elevated and erythrocyte membrane M (%) depressed, especially in those with a predisposition to allergies. PMID- 17461027 TI - [Economic evaluation of disability prevention programs for community-dwelling elderly secular trend analyses of medical and care expenses comparing participants and non-participants in the programs]. AB - OBJECTIVE: This study was conducted to evaluate disability prevention programs for community-dwelling elderly in terms of the cost/benefit balance. METHODS: Out of all residents aged 65 years and over who lived in Yoita Town, Niigata Prefecture, Japan (n = 1,673), 1,544 persons participated in the baseline survey conducted in November, 2000 (response rate, 92.2%). Based on the results, Yoita town then launched several kinds of disability prevention programs for community dwelling elderly, e.g., preventive programs for falls, dementia and homeboundness. The subjects for the present study included only those aged 70 years and over who responded to the baseline survey and were alive as of March 2004. During 2001 to 2003, 146 persons had participated at lease once in one of disability prevention programs (denoted as the participant group), whereas 846 persons had not participated in any of the programs (denoted as the non participant group). We compared medical and care expenses (sum of national and employment health insurance benefits, and long-term care insurance) between the two groups during 2000-2003, and determined whether participating in program affected subsequent medical and care expenses independent of key confounders [sex, age, and baseline medical and care expenses or health indicator (TMIG-Index of Competence or Generic Mobility Index)], using general linear models. RESULTS: The mean medical expenses per capita and per month slightly decreased over the period of the study in the participant group (51,606 yen for 2000 to 47,539 yen for 2003), while those in non-participant group steadily increased (41,888 yen, to 51,558 yen, respectively). During the same period, the mean care cost per capita and per month increased in both groups, but the increase was much more moderate in the participating group (507 yen to 5,186 yen vs. 8,127 yen to 27,072 yen for non-participant group). Summed cost reduction through the program participation accounted for 49 million yen during the three years (2001-2003). After adjustment for sex, age and baseline medical and care expenses or health indicator, it was estimated as 12 million yen per year. Given that the expenses for the disability prevention programs summed 2.3 million yen per year, the net benefit of disability prevention programs was estimated to be approx. 10 million yen per year. CONCLUSION: Disability preventive programs for the community dwelling elderly are economically efficient in terms of the cost/benefit balance. Future research is needed to examine how such programs lead to cost reduction. PMID- 17461028 TI - [The impact of changing critical values of the early epidemic detection system for infectious disease surveillance in Japan]. AB - INTRODUCTION: It is important to examine how critical values for initiation/termination affect the trend and frequency of epidemic/pre-epidemic warnings with the early epidemic detection system in Japan. Here we looked at the number of epidemic warning/pre-warning weeks and the influence of changing the criteria values for infectious diseases surveillance. METHODS: An epidemic warning is initiated if the number of cases per week per sentinel medical institution exceeds a critical value. A pre-warning for an epidemic is initiated if cases per week per sentinel medical institution exceed a critical value and there is a non-epidemic warning. To determine effects of the criteria values for epidemics/pre-epidemics for warning onset and termination, we set different values and compared the number of weeks of epidemic warning, the proportion of the total observed weeks. Also, pre-epidemic warning measurements were compared. Data from the infectious diseases surveillance system were analyzed from fiscal years 1999 to 2003. RESULTS: When the critical value for warning onset was lowered, the warning week started sooner and ended later. When the critical value was raised, the opposite occurred: the number of weeks with a warning status decreased. When the critical value was changed within a certain range, the number of weeks with a warning status became 0.5 to 2 times larger than those with the defined value. Similar trends were observed when the pre-warning was examined: the number of warning weeks was 0.4 to 2 times (for influenza and chickenpox) and 0.3 to 3 times (for measles and mumps) larger than those with the defined value. Except for pertussis and rubella, the proportion of warning weeks was approximately 5% for all diseases listed in the early epidemic detection system. In addition, there was no distinct issue with the critical values themselves. CONCLUSION: The present examination of linkage between trends and frequencies of epidemic warnings/ pre-warnings and the critical values in the early epidemic detection system of infectious disease surveillance in Japan confirmed the expected increase with lowering of the threshold. Except for pertussis and rubella, there was no distinct issue with the critical values themselves. PMID- 17461029 TI - [Epidemiological surveillance for viral pollution of rivers in Toyama Prefecture]. AB - OBJECTIVE: Virus pollution of three rivers in Toyama Prefecture was surveyed over a long period in order to predict and to prevent water-born infection. METHODS: Water samples were collected from three rivers (Itachi, Sembo, Oyabe), then concentrated and inoculated into cultured cells to isolate viruses. The survey at Itachi River was carried out 4 times: in 1979-1981, 1983-1985, 1993-1995, and 2002-2003. The surveys at Sembo and Oyabe Rivers were carried out twice (together with the 3rd and 4th surveys of Itachi River). RESULTS: 1) Various species of enteric viruses, i.e., poliovirus, human enteroviruses B (HEV-B), and reovirus were isolated from Itachi River. Since polioviruses were isolated at the same time as the oral vaccination of babies, these isolates appeared to be derived from vaccine strains. Consistently, isolates in the 3rd and the 4th surveys had only 0-2 base differences in the 480 or 474 nucleotide sequences of the VP3-VP1 region, compared with vaccine strains. The poliovirus detection rates, defined as the ratios of times viruses were detected to the total investigation times, were 33.3%, 41.7%, 2.1% and 0% for the Itachi River from the 1st to 4th surveys, respectively. The lowering between the 2nd and the 3rd surveys was significant (P< 0.001), this being associated with the widespread introduction of paper diapers for babies. The types of HEV-B were various and coincided well with those of prevalent clinical isolates. Reoviruses were frequently detected throughout the year in the 1st and the 2nd surveys, but fell in spring and summer in the 3rd and 4th surveys. 2) The types and the rates of viruses detected from Sembo and Oyabe Rivers were similar to those from Itachi River. CONCLUSIONS: The detection rates of poliovirus and reovirus in the river water during 2002 to 2003 were lower than during 1979 to 1981. This may be due to the improvement of sewerage system or sewerage and the increase in use of paper diapers for babies. The reason that the detection rate of HEV-B did not similarly decrease, and the fact that various types of HEV-B were isolated every year seemed to reflect the epidemic status of HEV-B among inhabitants. Thus, while virus pollution of river water has generally decreased, there is still a possibility of outbreak of water born infections, since rivers continue to be contaminated with various species of viruses. PMID- 17461030 TI - [p97ATPase-mediated biogenesis of the golgi, ER and nuclear envelope]. PMID- 17461031 TI - [Migration patterns of glia and glioma cells--their origins, pathways, and destinations]. PMID- 17461032 TI - Prognosis of patient with cardiopulmonary arrest transported to Kyushu University Hospital. AB - Three hundreds and sixty six patients with out-of-hospital cardiopulmonary arrest, transported to the Kyushu University Hospital from 2000 to 2006, were examined using the Utstein style in witnessed cardiogenic cardiopulmonary arrest patients. Also, we examined the influence on prognosis due to the difference in the treatment of airway control in out-of-hospital settings. Nineteen patients out of 78 witnessed cardiogenic out-of-hospital cardiopulmonary arrest patients were discharged alive and 11 were with a good prognosis. The number of cases where an initial electrocardiographic complex showed ventricular fibrillation or pulseless ventricular tachycardia was higher than formerly reported in Japan and was equal to the incidence reported in Europe and America. In addition, the survival discharge rate of patients with the ventricular fibrillation or pulseless ventricular tachycardia was higher than that previously reported in Japan and was similar to European and American results. Manual airway maintenance using a bag valve mask was more successful in terms of the survival discharge rate compared to the use of advanced airway devices. By the time course, collapse to cardiopulmonary resuscitation interval, collapse to initial defibrillation interval and collapse to the return of spontaneous circulation interval were shorter in the group discharged with a good prognosis, especially in the witnessed ventricular fibrillation or pulseless ventricular tachycardia patients corresponding to former reports. Most patients with a good prognosis resuscitated before arrival at the hospital. These results suggest the prehospital treatment is the critical point other than in-hospital treatment. PMID- 17461033 TI - Newly developed all in one EUS system: one cart system, forward-viewing optics type 360 degrees electronic radial array echoendoscope and oblique-viewing type convex array echoendoscope. AB - Most endosonographers use radial scanning instruments for diagnostic imaging, and use longitudinal scanning instruments primarily for endoscopic ultrasound-guided fine needle aspiration (EUS-FNA). The use of two separate instruments for radial and longitudinal scanning means 2 different echoendoscopes are required, each with its own dedicated US processing unit. Currently available electronic radial echoendoscopes and linear instruments made by the same company require the same other brand US unit. Furthermore, no forward-viewing optics type 360 degrees electronic radial echoendoscope currently exists. We have developed an all-in-one one cart EUS system that saves space and is available for both the forward viewing type 360 degrees radial electronic echoendoscope and the oblique-viewing type convex echoendoscope. These scopes have a transducer with variable frequency (5.0, 7.5, 10.0, 12.0 MHz) and color and power Doppler flow mapping capabilities. We performed a clinical development test for thirteen patients with sixteen lesions (Radial EUS on 8 lesions and EUS-FNA on 8 lesions) using this new EUS system. These new instruments provided satisfactory US and endoscopic images. The forward-viewing optics of the prototype enhanced intubation and instrument advancement. The radial scanning prototype provided an adequate diagnosis in 8 (100%) out of 8 lesions for EUS. The convex type achieved successful puncture in 8 (100%) out of 8 lesions and collection of adequate specimen for diagnosis of EUS-FNA in 4 (50%) out of 8 lesions. There were no complications in this series. This new system appears to be an attractive alternative for efficient EUS. PMID- 17461034 TI - [Evaluation of visual function in macular diseases]. PMID- 17461035 TI - [Study of the causes of poor visual prognosis in photodynamic therapy for age related macular degeneration]. AB - PURPOSE: To study the factors for poor visual outcome in photodynamic therapy(PDT) for age-related macular degeneration (AMD) patients with subfoveal choroidal neovascularization. MATERIALS AND METHODS: Medical charts of eighty five patients (88 eyes) who had AMD with subfoveal choroidal neovascularization were reviewed in the study. All patients were followed up by visual acuity assessment, angiography, and optical coherence tomography at least 6 months after PDT. RESULTS: Out of 11 cases of poor visual outcome, 7 cases showed cystoid macular edema (CME) before treatment and 4 cases showed massive subretinal hemorrhage and/or vitreous hemorrhage after treatment. Some cases with CME had a worse visual outcome than cases without CME. CONCLUSIONS: Some of the factors for poor visual outcome after PDT for AMD are the existence of CME before treatment and polypoidal choroidal vasculopathy (PCV). In some cases, PCV caused massive subretinal hemorrhage and/or vitreous hemorrhage after treatment. PMID- 17461036 TI - [Assessment of the quality of life in patients with age-related macular degeneration 1 year after photodynamic therapy]. AB - OBJECTIVE: To evaluate quality of life(QOL) in patients who underwent photodynamic therapy (PDT) for subfoveal choroidal neovascularization (CNV) of age-related macular degeneration (AMD), and to clarify factors influencing QOL. METHODS: 77 patients who had undergone PDT for subfoveal CNV of AMD were interviewed to evaluate QOL within 1 week before PDT, and 1 year after PDT. Subscale QOL scores were calculated and compared at these two time points. Factors related to QOL 1 year after PDT were evaluated. Factors included clinical findings in eyes which underwent PDT, condition of contralateral eyes, and family composition. QOL was also evaluated for patients whose visual acuity was improved (> or = logMAR 0.3)1 year after PDT. RESULTS: Subscale scores for activities associated with near vision (p = 0.03), mental health (p = 0.01), and role limitation (p = 0.04) improved 1 year after PDT. The only factors related to improvement in these subscale scores were low scores for each subscale before PDT. Mean logMAR visual acuity was 0.82 before PDT, and 0.72 at 1 year after PDT. Mean foveal thickness was 244 microm before PDT, and 189 microm at 1 year. Retinal detachment disappeared in 73% of the patients at 1 year. In 27 eyes with improved visual acuity at 1 year, three subscale scores, mental health (p = 0.01), role limitation (p = 0.03) and general vision (p = 0.04), showed improvement despite a mean foveal thickness of 165 microm and 93% disappearance of retinal detachment. CONCLUSION: QOL after PDT improved in association with improvements in clinical findings. However, QOL improvement was limited even when PDT was successful. PMID- 17461037 TI - [Sighting dominance in patients with macular disease]. AB - PURPOSE: To study sighting dominance by comparing macular disease patients undergoing surgical treatment with controls. METHODS: We studied visual acuity and sighting dominance in 92 macular disease patients, 27 of whom were assessed for both outcomes. We also studied visual acuity and sighting dominance in 412 controls. Sighting dominance was evaluated using the hole-in-card test. RESULTS: Among the controls, 70% showed right sighting dominance, and 30%, left sighting dominance. On the other hand, in patients with macular disease, right sighting dominance was demonstrated in 51%, and left in 49%; that is, 24% showed sighting dominance of the affected eye and 76%, of the fellow eye. During follow-up, sighting dominance of three of the 27 macular disease patients shifted from the affected eye to the fellow eye, which showed improvement in visual acuity. CONCLUSION: This study raises the possibility of sighting dominance shifting in patients with macular disease. There were differences among cases in the timing of the shift in sighting dominance, indicating that visual acuity may not be the only factor influencing sighting dominance. Further study is needed to confirm the factors contributing to sighting dominance. PMID- 17461038 TI - [Preoperatively administered flomoxef sodium concentration in aqueous humor]. AB - PURPOSE: We intravenously administered flomoxef sodium (FMOX) 0.5-3.5 hours before cataract surgery and measured the concentration of the agent in the aqueous humor to investigate its penetration into the aqueous humor and its efficacy in the prevention of postoperative endophthalmitis. METHODS: 56 patients who underwent cataract surgery were enrolled in this study. They received 1 g FMOX via a 20-minute intravenous drip beginning 0.5-3.5 hours before the operation. Aqueous humor was aspirated from the anterior chamber and assayed for FMOX concentration using high-performance liquid chromatography. RESULTS: The mean intraoperative FMOX concentrations in the patients' aqueous humor were 0.79 +/- 0.24 microg/ml (administered 3.5 hours before surgery)--1.47 0.79 microg/ml (administered 1.5 hours before surgery). These concentrations administered 0.5 3.0 hours before surgery sufficiently exceeded the minimum inhibitory concentration (MIC) 90 values against Staphylococcus epidermidis, Staphylococcus aureus and Propionibacterium acnes, but did not achieve the MIC90 values against Enterococcus faecalis and Pseudomonas aeruginosa. CONCLUSIONS: The FMOX concentrations in the aqueous humor sampling were adequate to kill bacteria in vitro. This drug may be efficacious in the prevention of postoperative endophthalmitis in patients undergoing cataract surgery. PMID- 17461039 TI - [Quantification of metamorphopsia in the course of branch retinal vein occlusion with M-CHARTS]. AB - PURPOSE: To evaluate visual function disorder in patients with branch retinal vein occlusion by quantifying the degree of metamorphopsia. DESIGN: prospective interventional case series. SUBJECTS AND METHODS: The subjects were 12 patients(12 eyes) whose fovea was involved with branch retinal vein occlusion within a month after noticing their symptoms. The best-corrected visual acuity, and vertical and horizontal metamorphopsia scores were obtained within one month, and at 3 months and 6 months after the onset of symptoms regardless of therapy. The metamorphopsia scores were measured with M-CHARTS. RESULTS: Although the average best-corrected visual acuity at 6 months was significantly better than that within a month of noticing symptoms, the average metamorphopsia scores did not improve after 6 months, and were not related to the average best-corrected visual acuity in either vertical or horizontal lines. CONCLUSIONS: The degree of metamorphopsia measured with M-CHARTS is an important parameter in addition to the best-corrected visual acuity for evaluating visual function in patients with branch retinal vein occlusion. PMID- 17461041 TI - [Lensectomy/lentectomy and vitritis/vitreitis]. PMID- 17461040 TI - [Peripherin/RDS gene mutation in a patient with choroidal neovascularization]. AB - PURPOSE: To diagnose an atypical retinal degenerative disease with choroidal neovascularization by means of gene diagnosis. CASE: A 47-year-old woman had good visual acuity at the first examination. She had scattered chorioretinal degeneration and pigmentation in the peripheral retina. There was a symmetrical visual field defect in the upper and temporal periphery in both eyes. Seven years later, choroidal neovascular membrane (CNV) developed in the fovea of her left eye and visual acuity deteriorated to 0.4 in this eye. Optical coherence tomography revealed type 2 CNV with minimal subretinal fluid. Fluorescein angiography showed very little leakage from the CNV. We used denaturing high performance liquid chromatography(DHPLC) to perform gene diagnosis and found a peripherin/RDS gene mutation of Gly167-Ser. CONCLUSION: Our case had moderate peripheral retinal degeneration with CNV. It is possible that cases like this tend to be misdiagnosed as AMD (age-related macular degeneration) or CNV with high myopia. Evaluation of the gene mutation was useful for diagnosis in this case. PMID- 17461042 TI - [Suggestion for the registry of retinoblastoma in Japan]. PMID- 17461043 TI - [A review 30. Clinical aspects of granulomatous uveitis in Japan]. PMID- 17461044 TI - Confidence limits and sample size for determining nonhost status of fruits and vegetables to tephritid fruit flies as a quarantine measure. AB - Quarantine measures including treatments are applied to exported fruit and vegetable commodities to control regulatory fruit fly pests and to reduce the likelihood of their introduction into new areas. Nonhost status can be an effective measure used to achieve quarantine security. As with quarantine treatments, nonhost status can stand alone as a measure if there is high efficacy and statistical confidence. The numbers of insects or fruit tested during investigation of nonhost status will determine the level of statistical confidence. If the level of confidence of nonhost status is not high, then additional measures may be required to achieve quarantine security as part of a systems approach. Certain countries require that either 99.99 or 99.9968% mortality, as a measure of efficacy, at the 95% confidence level, be achieved by a quarantine treatment to meet quarantine security. This article outlines how the level of confidence in nonhost status can be quantified so that its equivalency to traditional quarantine treatments may be demonstrated. Incorporating sample size and confidence levels into host status testing protocols along with efficacy will lead to greater consistency by regulatory decision-makers in interpreting results and, therefore, to more technically sound decisions on host status. PMID- 17461045 TI - Comparison of parasitic mites in Russian-hybrid and Italian honey bee (Hymenoptera: Apidae) colonies across three different locations in North Carolina. AB - The most economically important parasites of honey bee, Apis mellifera L. (Hymenoptera: Apidae), colonies are the parasitic mites Varroa destructor Anderson & Trueman and Acarapis woodi (Rennie). Research has shown that mite tolerant stocks are effective means to reduce mite infestations within colonies, but it is unclear whether the stocks available commercially are viable means of mite control because they are likely to be genetic hybrids. We compared colonies of a standard commercial stock ("Italian") with those of a commercially purchased mite-tolerant stock ("Russian") for their levels of varroa and "tracheal" mites (A. woodi) over the course of 2 yr in three different geographic locations. We were unable to detect significant infestations of tracheal mites; thus, we were unable to adequately compare the stocks for their tolerance. In contrast, we found significant differences in the levels of varroa mites within and among colonies located across the three different study sites for both years. By the end of the first year, we found statistically significant differences between the stocks in varroa mite intensity (mites per adult bee), such that Russian-hybrid colonies tended to have a significantly lower proportion ofparasitized adult bees than Italian colonies. In the second year, we found statistically significant differences between the stocks in varroa mite load (daily mite drop), such that Russian-hybrid colonies tended to have lower total numbers of mites than Italian colonies. These findings suggest that beekeepers may benefit by incorporating commercially purchased mite-tolerant stocks into their existing integrated pest management programs. PMID- 17461046 TI - Flight activity of USDA-ARS Russian honey bees (Hymenoptera: Apidae) during pollination of lowbush blueberries in Maine. AB - Flight activity was compared in colonies of Russian honey bees, Apis mellifera L. (Hymenoptera: Apidae), and Italian bees during commercial pollination of lowbush blueberries (principally Vaccinium angustifolium Aiton) in Washington Co., ME, in late May and early June in 2003 and 2004. Colonies of the two stocks were managed equally in Louisiana during autumn through early spring preceding observations in late spring each year. Resulting average populations of adult bees and of brood were similar in colonies of the two bee stocks during pollination. Flight during pollination was monitored hourly on 6 d each year by counting bees exiting each colony per minute; counts were made manually with flight cones on 17 colonies per stock in 2003 and electronically with ApiSCAN-Plus counters on 20 colonies per stock in 2004. Analysis of variance showed that temperature, colony size (population of adult bees or brood), and the interaction of these effects were the strongest regulators of flight activity in both years. Russian and Italian bees had similar flight activity at any given colony size, temperature, or time of day. Flight increased linearly with rising temperatures and larger colony sizes. Larger colonies, however, were more responsive than smaller colonies across the range of temperatures measured. In 2003, flight responses to varying temperatures were less in the afternoon and evening (1500-1959 hours) than they were earlier in the day. Russian colonies had flight activity that was suitable for late spring pollination of lowbush blueberries. PMID- 17461047 TI - Sterile insect technique and Mediterranean fruit fly (Diptera: Tephritidae): assessing the utility of aromatherapy in a Hawaiian coffee field. AB - The sterile insect technique (SIT) is widely used in integrated programs against tephritid fruit fly pests, particularly the Mediterranean fruit fly, Ceratitis capitata (Wiedemann) (Diptera: Tephritidae). Unfortunately, the mass-rearing procedures inherent to the SIT often lead to a reduction in the mating ability of the released males. One potential solution involves the prerelease exposure of males to particular attractants. In particular, exposure of male Mediterranean fruit flies to ginger, Zingiber officinale Roscoe, root oil (GRO) has been shown to increase mating success in laboratory and field cage trials. Here, we describe a field experiment that compares the level of egg sterility observed in two Hawaiian coffee, Coffea arabica L., plots, with GRO-exposed, sterile males released in one (treated) plot and nonexposed, sterile males released in the other (control) plot. Once per week in both plots over a 13-wk period, sterile males were released, trap captures were scored to estimate relative abundance of sterile and wild males, and coffee berries were collected and dissected in the laboratory to estimate the incidence of unhatched (sterile) eggs. Data on wild fly abundance and the natural rate of egg hatch also were collected in a remote area that received no sterile males. Despite that sterile:wild male ratios were significantly lower in the treated plot than in the control plot, the incidence of sterile eggs was significantly higher in the treated plot than in the control plot. Correspondingly, significantly higher values of Fried's competitiveness index (C) were found, on average, for treated than control sterile males. This study is the first to identify an association between the GRO "status" of sterile males and the incidence of egg sterility in the field and suggests that prerelease, GRO exposure may represent a simple and inexpensive means to increase the effectiveness of Mediterranean fruit fly SIT programs. PMID- 17461048 TI - Multiple orifice distribution system for placing green lacewing eggs into verticel larval rearing units. AB - Green lacewings are widely used biological control agents for various insect pests. To meet the needs of growers, green lacewings are being mass-reared commercially around the world. A common salt shaker has been used regularly to distribute eggs into Verticel lacewing larval rearing units. This technique is time consuming and inefficient because the number of eggs distributed in each cell is inconsistent. The multiple orifice distribution (MOD) system described here greatly improved egg distribution efficiency by increasing the percentage of Verticel cells containing the desired one to four eggs per cell (i.e., 40.8 and 52.1% by using salt shaker method versus 61.9% by using the MOD system). This mechanical system significantly reduced the labor and time involved in the process and would cost under $3500. In addition, this new system could be modified for distribution of other insect eggs. PMID- 17461049 TI - Horizontal transmission of Beauveria bassiana in Anastrepha ludens (Diptera: Tephritidae) under laboratory and field cage conditions. AB - The virulence of two products of the fungus Beauveria bassiana (LCPP and Bassianil) on adult Anastrepha ludens (Loew) (Diptera: Tephritidae) and their effect on the mating performance of infected males was evaluated in laboratory and field cage tests. The horizontal transmission capacity of the fungus during copulation or attempted copulation also was quantified using inoculated males as well as the impact of infection on female fecundity and longevity. Both fungal products were found to be highly virulent (LCPP, mortality = 98.7% at 1 x 10(8) conidia per ml, LT50 = 4.20 d, LC50 = 9.35 x 10(5) conidia per ml; Bassianil, mortality = 99.3% at 1 X 10(8) conidia per ml, LT50 = 4.04 d, LC50 = 2.69 x 10(7) conidia per ml). Mating success of inoculated males was not affected compared with the control group during the 3 d postinoculation. Horizontal transmission to females during the first day was 80.6 and 84.3% through mating and 15.4 and 21.6% through attempts to mate and contact during courtship for the LCPP and Bassianil products, respectively. The fertility of infected females was notably reduced, and longevity did not extend beyond 15 d. Our results suggest the possibility of using sterile flies as fungus vectors in sterile insect technique programs, but the potential benefits and shortcomings of this approach require further investigation. PMID- 17461050 TI - Parasitic hymenoptera fauna on agromyzidae (diptera) colonizing weeds in ecological compensation areas in northern Italian agroecosystems. AB - Parasitoids (Hymenoptera) associated with agromyzid leafminers (Diptera: Agromyzidae) were studied in three rural farms located in northern Italy. The parasitoids were reared from mined foliage of weeds growing in field margins. We reared 998 Hymenoptera specimens, representing five families, 23 genera, and 53 species, from leafminers infesting weeds. Eulophidae was the most abundant family (67.64%), followed by Braconidae (28.86%), Eucoilinae (1.40%), Tetracampidae (1.40%), and Pteromalidae (0.7%). Braconids was the most species rich family, accounting for 28 species; eulophids were represented by 19 species, pteromalids by four species, and eucoilins and tetracampids by one species each. The dominant parasitoid was the eulophid Pediobius metallicus (Nees), representing 18.17% of the total, followed by Diglyphus isaea (Walker) (12.73%), and Neochrysocharis formosa (Westwood) (10.82%). The most abundant braconid parasitoid was Dacnusa maculipes Thomson (9.62%). More than 80% of parasitoids were recovered from 10 plant species: Cirsium arvense (L.) Scopoli, Plantago lanceolata L., Sonchus asper (L.) Hill, Papaver rhoeas L., Picris echioides L., Lactuca serriola L., Myagrum perfoliatum L., Ranunculus velutinus Tenore, Arctium lappa L., and Medicago sativa L. The retention and the management of wild plants within field margins can be crucial tools to enhance the populations of biological control agents of agromyzids and to conserve rare parasitic wasp species. PMID- 17461051 TI - Chlorpyrifos immersion to eliminate third instars of Japanese beetle (Coleoptera: Scarabaeidae) in balled and burlapped trees and subsequent treatment effects on red maple. AB - This study examined chlorpyrifos immersion of balled and burlapped (B&B) nursery trees for elimination of third instars of Japanese beetle, Popillia japonica Newman (Coleoptera: Scarabaeidae), and for phytotoxicity on red maple, Acer rubrum L. Trees were harvested as 45- and 60-cm-diameter B&B and immersed in chlorpyrifos at U.S. Domestic Japanese Beetle Harmonization Plan rate (0.24 kg active ingredient [AI/100 liters) or lower rates of 0.015, 0.03, 0.06, and 0.12 kg (AI)/100 liters. The 0.03, 0.06, and 0.24 kg (AI) rates provided 100% control of Japanese beetle grubs in both 45- and 60-cm B&B. The 0.015 and 0.12 kg (AI) chlorpyrifos rates were 100% effective in three tests. However, in another test, 0.015 and 0.12 kg (AI) chlorpyrifos treatments had four (93% control) and one (98% control) grubs recovered, respectively. Root ball soils consisted of loam, silt loam, or clay loam texture classifications. Trunk diameter and internode growth of red maple harvested as 45-cm B&B decreased linearly with increasing chlorpyrifos dip rate during the first year, but effects were unapparent in the second year. Chlorpyrifos rates had no measurable impact on growth of red maples harvested as 60-cm B&B. No visual phytotoxicity symptoms were detected for chlorpyrifos rate or root ball size treatments. In conclusion, results support lowering the U.S. Domestic Japanese Beetle Harmonization Plan chlorpyrifos dip rate for category 2 states to at least 0.03 kg (AI) for B&B diameters < or =60 cm. Chlorpyrifos rates < 0.24 kg (AI) will lower cost, reduce worker exposure, and lessen potential environmental contamination. PMID- 17461052 TI - Assessment of risk of introduction of Cylas formicarius elegantulus (Coleoptera: Brentidae) into weevil-free areas in the southern United States. AB - We assessed the risk associated with introduction of sweet potato weevil, Cylas formicarius elegantulus (Summers) (Coleoptera: Brentidae), from infested areas to noninfested areas via shipment of sweet potato, Ipomoea batatas (L.), roots within the southern United States. Our study quantifies the effectiveness of risk mitigation procedures of sweet potatoes before shipment in relation to introduction of the weevil. The risk assessment relied on literature and expert information to determine appropriate parameters. Using a computational model, Monte Carlo simulations were conducted to estimate the likelihood of introduction of sweet potato weevil. Risk management options were incorporated and the risk analyses were performed to assess how the risk could be reduced. The study found the risk of introduction of the weevil for both domestic shipment and imports of sweet potatoes into new areas within the southern United States to be low. Sensitivity analysis was performed to assess model stability and the impact of parameter changes. Based on the sensitivity analysis, the most critical input was the postharvest mitigation, followed by the number of weevils per ton of sweet potatoes. We concluded that maintaining mitigations with monitoring in conjunction with public education to stop illegal transport of sweet potatoes and alternate hosts would significantly reduce the risk of introduction. PMID- 17461053 TI - Wheat germ oil and its effects on a liquid larval rearing diet for oriental fruit flies (Diptera: Tephritidae). AB - Wheat germ oil was added to a larval liquid diet for rearing Bactrocera dorsalis (Hendel) (Diptera: Tephritidae) to optimize fruit fly quality. Effects of various concentrations of wheat germ oil at 0.04, 0.07, 0.15, 0.30, and 0.66% and their possible mode of action were evaluated. Results suggest that addition of wheat germ oil does not affect pupal weight, larval developmental period, adult emergence, mating ability, or peak time for egg production. But there was a significant increase in pupal recovery, percentage of adult fliers, egg production, or egg hatch for larvae fed the diet with wheat germ oil compared with those reared on the liquid diet without wheat germ oil. The increase in egg hatch and fliers was dose dependent. Therefore, addition of wheat germ oil to fruit fly rearing diet is a novel way to improve fruit fly quality, especially in egg hatch, fliers, egg production, and pupal recovery. PMID- 17461054 TI - Effects of bacillus thuringiensis transgenic corn on corn earworm and fall armyworm (Lepidoptera: Noctuidae) densities. AB - We examined 17 pairs of near-isogenic hybrids of Bacillus thuringiensis (Bt) (176, Mon810, and Bt11) and non-Bt corn, Zea mays L., to examine the effects of Bt on larval densities of Helicoverpa zea (Boddie) and Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctuidae) during 2 yr. During ear formation, instar densities of H. zea and S. frugiperda were recorded for each hybrid. We found that H. zea first, second, and fifth instar densities were each affected by Mon810 and Bt11 Bt corn but not by 176 corn. Surprisingly, first and second instars were found in higher numbers on ears of Mon810 and Bt11 corn than on non Bt corn. Densities of third and fourth instars were equal on Bt and non-Bt hybrids, whereas densities of fifth instars were lower on Bt plants. S. frugiperda larval densities were only affected during 1 yr when second, and fourth to sixth instars were lower on ears of Mon810 and Bt11 hybrids compared with their non-Bt counterparts. Two likely explanations for early instar H. zea densities being higher on Bt corn than non-Bt corn are that (1) Bt toxins delay development, creating a greater abundance of early instars that eventually die, and (2) reduced survival of H. zea to later instars on Bt corn decreased the normal asymmetric cannibalism or H. zea-S. frugiperda intraguild predation of late instars on early instars. Either explanation could explain why differences between Bt and non-Bt plants were greater for H. zea than S. frugiperda, because H. zea is more strongly affected by Bt toxins and more cannibalistic. PMID- 17461055 TI - Monitoring populations of saddled prominent (Lepidoptera: Notodontidae) with pheromone-baited traps. AB - Field trials with three types of pheromone traps were performed in eight northern hardwood stands in northern New York state to develop a population-monitoring tool for the saddled prominent, Heterocampa guttivitta (Walker) (Lepidoptera: Notodontidae). Lure specificity and the relationship between pheromone trap catch and subsequent egg density were examined. A study of moth emergence in relation to temperature was designed to determine whether moth activity throughout the flight season can be predicted using a growing degree-day (DD) model. Pherocon 1C wing traps were significantly more effective than the green Unitrap bucket style. Catch was not affected by position when traps were > or =20 m from an opening (road), and lures were specific to saddled prominent. Lure specificity was examined using green Multipher bucket traps, which effectively attracted and held moths. In the first year of the study, number of viable eggs per 10 leaf clusters was significantly correlated (r2 = 0.59) with average moth catch/trap in pheromone-baited Pherocon traps. When differences in stand density (basal area) and relative abundance of sugar maple (percentage of total stems per hectare), the principle host, were accounted for, the multiple regression model also was significant and r2 = 0. 83. Neither model, however, was significant the second year. Using a base temperature of 5.5 degrees C and on-site temperature data, the peak of moth flight occurred at 316 +/- 8 DD and end of flight occurred at 533 +/ 9 DD. PMID- 17461056 TI - Impact of temperature on plum curculio (Coleoptera: Curculionidae) responses to odor-baited traps. AB - In 2005, captures of overwintered adult plum curculios, Conotrachelus nenuphar (Herbst) (Coleoptera: Curculionidae), in standard black masonite pyramid traps deployed in apple (Malus spp.) orchards from half-inch green until fruit reached 7 mm and baited with known attractants did not result in significant captures compared with unbaited traps as they had in 2003 and 2004. These baits included the synthetic aggregation pheromone, grandisoic acid (GA) alone, a six-component synthetic host plant volatile combination (6-Tree) identified from foliar and woody tissues of a Stanley plum tree in combination with GA (6-Tree+GA), and the synthetic fruit volatile benzaldehyde (BEN) in combination with GA (BEN+GA). In 2005, the average daily temperature was below 13 degrees C, much cooler than in 2003 and 2004. We hypothesized that plum curculio could not discriminate between baited and unbaited traps because of reduced release rates of odor-bait stimuli due to their temperature-driven release system. From data collected from 2003 to 2005, we found that plum curculio captures in traps baited with GA alone, 6 Tree+GA, and BEN+GA were significantly related to temperature. We created a predictive model to determine the level of activity, i.e., trap captures in baited traps compared with unbaited traps, we would expect to observe at a particular temperature for these same odor stimuli. Our models predicts that at temperatures between approximately 11 to 13 degrees C we would expect to see no difference between captures in baited and unbaited traps. For captures in odor baited traps to reach twice those in unbaited traps, our model predicts that temperatures must reach 19.2 degrees C for GA alone, 18.5 degrees C for 6 Tree+GA, and 15.8 degrees C for BEN+GA. PMID- 17461057 TI - Thermal requirement and development of Liriomyza sativae (Diptera: Agromyzidae) on cucumber. AB - The effect of temperature on developmental rate of vegetable leafminer, Liriomyza sativae Blanchard (Diptera: Agromyzidae), was determined at seven constant temperatures (10, 15, 20, 25, 30, 35, and 40 degrees C) on cucumber, Cucumis sativus L. 'Negin'. The total developmental period (oviposition to adult emergence) decreased with increasing temperature, although no development occurred at 10 and 40 degrees C. Using the linear model, the estimated lower temperature threshold for the egg, larval, pupal, and entire developmental period was 9.20, 9.75. 11.01, and 10.20 degrees C and the effective accumulative temperatures for these stages were 64.10, 81.97, 106.38, and 250 degree-days (DD), respectively. Data also were fitted to nonlinear temperature-dependent models. Evaluation of the models was based on fit to data, number and biological value of the fitted coefficient, number of measurable parameters, and accuracy of the estimation of the thresholds. Conclusively, linear and Briere models are recommended as the most efficient for the description of temperature-dependent development of L. sativae. Temperature-based developmental data can be used to predict occurrence, number of generations, and possibly population dynamics. PMID- 17461058 TI - Temperature-dependent variation in toxicity of insecticides against Earias vitella (Lepidoptera: Noctuidae). AB - The toxicity of synthetic pyrethroids was found to be negatively correlated with temperature, whereas contrasting correlation was observed with the toxicity of organophosphorous compounds chlorpyriphos and quinalphos, which was most toxic at higher temperature. A similar phenomenon was observed in endosulfan at higher temperature and humidity combination. The insecticide molecules indoxacarb and spinosad were effective among the insecticides tested. Indoxacarb was effective at lower temperature, and spinosad was effective at all the temperature and relative humidity combinations with minor difference in LD50 values. During both the years, however, the levels of resistance were higher in second year compared with previous year. PMID- 17461059 TI - Evaluation of substituted oxime ethers for growth regulatory activity against Spodoptera litura (Lepidoptera: Noctuidae). AB - Insect growth regulator (IGR) activity of 52 substituted oxime ethers was evaluated against an important polyphagous lepidopteran pest, Spodoptera litura (F.) (Lepidoptera: Noctuidae). Several compounds produced symptoms comparable to exogenously applied juvenile hormone. Maximum ICR activity was exhibited by 4' (2,6,6-trimethyl-2-cyclohexen-1-yl)-3'-buten-2'(E)-ketoxime-N-O-alkyl ether with an ED50 (morphological) of 40 microg g(-1) body weight, compared with 20 microg g(-1) of juvenile hormone (JH) III. Two more compounds, namely, 4'-(2,6,6 trimethyl-2-cyclohexen-1-yl)-3'-buten-2'(Z)-ketoxime-N-O-methyl propyl ether (ED50 of 192 microg g(-1)) and 4'-(2,6,6-trimethyl-1-cyclohexen-1-yl)-3'-buten 2'(E)-ketoxime-N-O-pentyl ether (ED50 of 380 microg g(-1)) showed considerable IGR activity, whereas 4'-(2,6,6-trimethyl-2-cyclohexen-1-yl)-3'-buten-2'(E) ketoxime-N-O-pentyl ether was found to be acutely toxic to larvae (ED50 of 268 microg g(-1)). Three compounds used in this study also were synergized by piperonyl butoxide. Synergistic ratios ranged from 1.330 to 4.605. No significant ovicidal activity was obtained. PMID- 17461060 TI - A kairomone based attract-and-kill system effective against alfalfa looper (Lepidoptera: Noctuidae). AB - A chemical lure derived from flowers that are visited by moths attracts male and female alfalfa loopers, Autographa californica (Speyer) (Lepidoptera: Noctuidae). This feeding attractant is dispensed from polypropylene bottles that provide controlled release for several weeks. A killing station was tested in the laboratory, in a screenhouse, and in the field in combination with this lure as an "attract-and-kill" system. Starved alfalfa looper adults (moths) were strongly attracted to the attract-and-kill station in a flight tunnel, and 90.9% of female moths and 87.6% of male moths that contacted the station died. In commercial fields of alfalfa hay, female moths captured in monitoring traps were reduced by 80-93% in plots receiving 125 attract-and-kill stations per hectare. In screenhouse trials using two attract-and-kill stations per screenhouse, oviposition on potted lettuce plants by starved female alfalfa looper moths was reduced by 98.5%. Moths were less likely to be attracted to lures when provided sugar before flight tunnel assays, and oviposition by fed moths was much less affected by attract-and-kill stations in screenhouse trials, compared with starved moths. This method has potential as a means to manage alfalfa looper populations in vegetable and other agricultural crops. However, consideration must be given to competing food and odor sources in the field. PMID- 17461061 TI - Distribution of imidacloprid residues in different parts of rice plants and its effect on larvae and adult females of Chilo suppressalis (Lepidoptera: Pyralidae). AB - The distribution of imidacloprid residue in different parts of rice, Oryza sativa L. (Graminales: Poaceae), plants was investigated for three rice varieties. Changes in larval hormones in Chilo suppressalis (Walker) (Lepidoptera: Pyralidae) that fed on imidacloprid-treated plant sources and the fecundity of resultant adult females (moths) also were determined. No significant differences in imidacloprid residue were detected in rice stem, leaves, and the unexpanded new leaf 7 d after treatment except in the rice variety Fengyouxiangzhan, in which residue content in leaves and the unexpanded new leaf was significantly lower than that in the stem. The percentage of reduction of imidacloprid residue after treatment varied with rice variety and plant parts. The greatest reduction in the three plant parts was shown in Wujing 15 compared with the other two varieties. The decomposition rate of imidacloprid residue in Wujing 15 was greater in leaves and unexpanded new leaf than in the stem, whereas in Wuyunjing 7 it was lower in leaves than in the unexpanded new leaf and stem. In Fengyouxiangzhan, the decomposition rate of imidacloprid in the stem increased 14 d after treatment. Juvenile hormone III and molting hormone titers in fourth instar larvae that had fed on rice plants treated with imidacloprid since the neonate stage were significantly higher than those in larvae fed on untreated plants (control). Juvenile hormone titer in larvae that had fed on Zizamia lalifolia Turcz galls treated with imidacloprid and the fecundity of the female moths that developed from such larvae were also significantly higher than that on the control counterparts. These results are discussed in relation to the impact of imidacloprid on nontarget insects and pest management. PMID- 17461062 TI - Development of a satellite-based hazard rating system for Dendroctonus frontallis (Coleoptera: Scolytidae) in the Ouachita Mountains of Arkansas. AB - The southern pine beetle, Dendroctonus frontalis Zimmermann (Coleoptera: Scolytidae), is the most damaging forest insect pest of pines (Pinus spp.) throughout the southeastern United States. Hazard rating schemes have been developed for D. frontalis, but for these schemes to be accurate and effective, they require extensive on-site measurements of stand attributes such as host density, age, and basal area. We developed a stand hazard-rating scheme for several watersheds in the Ouachita Highlands of Arkansas based upon remotely sensed data and a geographic information system. A hazard model was developed using stand attributes (tree species, stand age and density, pine basal area, and landform information) and was used to establish baseline hazard maps for the watersheds. Landsat 7 ETM+ data were used for developing new hazard maps. Two dates of Landsat imagery were used in the analyses (August 1999 and October 1999). The highest correlations between hazard rating scores and remotely sensed variables from either of the dates included individual Landsat 7 ETM+ bands in the near- and mid-infrared regions as well as variables derived from various bands (i.e., Tasseled cap parameters, principal component parameters, and vegetation indices such as the calculated simple ratio and normalized difference vegetation index). Best subset regression analyses produced models to predict stand hazard to southern pine beetle that consisted of similar variables that resembled but were more detailed than maps produced using inverse distance weighted techniques. Although the models are specific for the study area, with modifications, they should be transferable to geographically similar areas. PMID- 17461063 TI - Greenhouse whitefly (Homoptera: Aleyrodidae) dispersal under different UV-light environments. AB - The greenhouse whitefly, Trialeurodes vaporariorum (Westwood) (Homoptera: Aleyrodidae), is known to respond to UV light (UV). Field studies were conducted to improve our understanding of the behavioral effects and practical implications of using UV-blocking plastic films for the control of whitefly. Adult whiteflies were released in outdoor-located choice-chamber experiments with compartments clad with a range of films that transmitted incident UV to different extents. In release-recapture experiments, a very small proportion of the whiteflies recovered had dispersed into compartments where the entire UV spectrum was blocked, whereas the major proportion preferred compartments with UV. Compartments clad with films that blocked UV below 375 nm attracted significantly more whiteflies than films that blocked UV below 385 nm, whereas the absorption of UV wavelengths above 385 nm did not show any further effect on whitefly numbers. A reduction in the side cladding of the compartments by >20% significantly reduced the advantage of using UV-blocking films. Adult whitefly did not discriminate between direct- and diffused-light environments, as long as the UV-absorbing properties of the films were equivalent. Whitefly dispersal was influenced by the time of the day when adult whitefly were released, with a higher proportion of whitefly avoiding compartments clad with UV-blocking films, at times of the day when light intensities were higher. The future use of UV blocking films as a potentially highly effective component of integrated pest management systems for the control of whitefly is discussed. PMID- 17461064 TI - Biology and control of the raspberry crown borer (Lepidoptera: Sesiidae). AB - This study explored the biology of raspberry crown borer, Pennisetia marginata (Harris) (Lepidoptera: Sesiidae), in Arkansas and the optimum timing for insecticide and nematode applications. The duration of P. marginata's life cycle was observed to be 1 yr in Arkansas. Insecticide trials revealed that bifenthrin, chlorpyrifos, imidacloprid, metaflumizone, and metofluthrin efficacy were comparable with that of azinphosmethyl, the only labeled insecticide for P. marginata in brambles until 2005. Applications on 23 October 2003 for plots treated with bifenthrin, chlorpyrifos, and azinphosmethyl resulted in >88% reduction in larvae per crown. Applications on 3 November 2004 of metaflumizone, metofluthrin, and bifenthrin resulted in >89% reduction in larvae per crown. Applications on 7 April 2005 for metofluthrin, imidacloprid, bifenthrin, metaflumizone, and benzoylphenyl urea resulted in >64% reduction in the number of larvae per crown. Applications on 6 May 2004 did not reduce larval numbers. The optimum timing for treatments was found to be between October and early April, before the larvae tunneled into the crowns of plants. Applying bifenthrin with as little as 468 liters water/ha (50 gal/acre) was found to be as effective against larvae as higher volumes of spray. Nematode applications were less successful than insecticides. Nematode applications of Steinernemafeltiae, Steinernema carpocapsae, and Heterorhabditis bacteriophora reduced larvae counts per plant by 46, 53, and 33%, respectively. PMID- 17461065 TI - Comparison of sex pheromone traps for monitoring pink hibiscus mealybug (Hemiptera: Pseudococcidae). AB - The pink hibiscus mealybug, Maconellicoccus hirsutus (Green) (Hemiptera: Pseudococcidae), is a highly polyphagous pest that invaded Florida in 2002 and has recently been reported from several locations in Louisiana. Although identification of its sex pheromone in 2004 improved monitoring capabilities tremendously, the effectiveness and efficiency of different pheromone trap designs for capturing males has not been evaluated. We deployed green Delta, Pherocon IlB, Pherocon V, Jackson, and Storgard Thinline traps in Homestead, FL, and compared the number of male M. hirsutus captured per trap, the number captured per unit of trapping surface area, the amount of extraneous material captured, and the time taken to count trapped mealybugs. Pheromone-baited traps with larger trapping surfaces (green Delta, Pherocon IIB, and Pherocon V) captured more males per trap than those with smaller surfaces (Jackson and Storgard Thinline), and fewest males were captured by Storgard Thinline traps. However, Jackson traps captured as many or more males per square centimeter of trapping surface as those with larger surfaces, and the time required to count males in Jackson traps was significantly less than in green Delta, Pherocon IIB, and Pherocon V traps. Although all trap designs accumulated some debris and nontarget insects, it was rated as light to moderate for all designs. Based on our measures of effectiveness and efficiency, the Jackson trap is most suitable for monitoring M. hirsutus populations. Additionally, unlike the other traps evaluated, which must be replaced entirely or inspected in the field and then redeployed, only the sticky liners of Jackson traps require replacement, enhancing the efficiency of trap servicing. PMID- 17461066 TI - Effects of azadirachtin, abamectin, and spinosad on sweetpotato whitefly (Homoptera: Aleyrodidae) on tomato plants under laboratory and greenhouse conditions in the humid tropics. AB - Direct and residual toxicity of NeemAzal-T/S (azadirachtin), Success (spinosad), and abamectin was tested against different life stages of sweetpotato whitefly, Bemisia tabaci (Gennadius) (Homoptera: Aleyrodidae), under air-conditioned laboratory conditions and in a tropical net greenhouse. NeemAzal-T/S and abamectin deterred the settling of adults on tomato, Lycopersicon esculentum Mill (Solanaceae), plants and consequently reduced egg deposition. No such effect was detected for Success. All three pesticides influenced egg hatch. Effects of NeemAzal-T/ S were significantly altered if applied to different-aged eggs (1, 3, and 5 d old). In contrast, abamectin-treated eggs failed to hatch at any given age class. All three products caused heavy mortality of the three nymphal stages of B. tabaci, with the first instars being most susceptible, abamectin-treated nymphs died within 24 h postapplication. In contrast, 100% nymphal mortality with NeemAzal-T/S and Success was reached 6-9 d postapplication. abamectin caused 100% immature mortality at all residue ages (1, 5, 10, and 15 d) in the laboratory and greenhouse as well. Persistence of Success was comparably high in the laboratory, but in the greenhouse a faster decline of activity was evident by increased egg deposition, egg hatch, and reduced rates of immature mortality. Toxicity of NeemAzal-T/S however gradually declined under greenhouse conditions with time (5 d) postapplication. The findings are discussed within the context of integrated management of whitefly under protected cultivation in the humid tropics. PMID- 17461067 TI - Effects of orchard host plants (apple and peach) on development of oriental fruit moth (Lepidoptera: Tortricidae). AB - Studies were designed to examine the effects of host plants (apple, Malus domestica Borkh., and peach, Prunus persica L.) on the development of oriental fruit moth, Grapholita molesta (Busck) (Lepidoptera: Tortricidae). Oriental fruit moth larvae developed faster on peach than on apple, both on fruit as well as on growing terminal shoots. On fruit, these differences were shown to cause significant changes in both the rate (approximately 20-60 degree-days earlier emergence on peach than on apple) and patterns of adult emergence among several cultivars of peaches and apples. Slopes of female emergence plots varied by host in 2003, with emergence occurring over a longer period on peach cultivars than on apple cultivars (with one exception). Slopes of male emergence curves did not differ by cultivar in 2003. These host-driven effects could impact the efficacy of traditional pest management approaches and probably complicate efforts to predictively model G. molesta populations in mixed cultivar orchards. Such developmental effects may help to explain previously observed differences in patterns of pheromone trap captures in peach versus apple orchards. Host associated effects should be incorporated into future models to develop more realistic predictive tools and thus improve integrated pest management efforts. PMID- 17461068 TI - Residual effects of imidacloprid on Japanese beetle (Coleoptera: Scarabaeidae) oviposition, egg hatch, and larval viability in turfgrass. AB - Preventive control of turf-infesting scarabaeid grubs by neonicotinoid insecticides is presumed to mainly result from residues killing first instars in the soil. The extent to which sublethal behavioral effects or intoxication of other life stages contribute to such control is poorly known. We tested whether Japanese beetle, Popillia japonica Newman, females lay fewer eggs in turf treated with imidacloprid (Merit 75 WP) or an imidacloprid-bifenthrin combination (Allectus GC SC), and whether exposure to those residues in thatch and soil reduces their survival and subsequent ability to feed or take flight. Effects of imidacloprid residues on egg hatch and viability of successive larval instars also were studied. In two sets of choice tests, 68 and 82% fewer eggs were laid in Kentucky bluegrass with Allectus residues than in controls. When females were confined in treated turf, however, neither insecticide consistently reduced their fecundity or affected depth at which eggs were laid, although exposure to fresh Allectus residues reduced the beetles' subsequent viability. Imidacloprid residues up to 2 ppm in soil did not affect egg viability or days to hatch, but they killed neonates soon after eclosion. Imidacloprid curatively applied at label rate (0.34 kg active ingredient/ha) reduced weight gain, burrowing capability, frass production, and survival of second and third instars in turfgrass cores, with high mortality within 30 d. Intoxication and behavioral impairment of third instars also occurred in autumn field trials. Our data suggest that imidacloprid has greater activity against late instars than is generally appreciated. PMID- 17461069 TI - Development of bait stations for fruit fly population suppression. AB - The application of insecticides is an essential component for eradication or management of fruit fly pests. Impact on nontarget organisms and public rejection of areawide pesticide applications have been major concerns in managing these programs. Bait stations have been proposed as alternative treatments in areas where broadcast insecticides are not acceptable. In this study, we defined bait stations as discrete containers of attractants and toxins, which are targeted at specific pests. Tests were carried out using the Mexican fruit fly, Anastrepha ludens (Loew), as the experimental insect. Our first bait station design was a sheet of sponge material fastened to a plastic peaked cover. Liquid bait consisting of protein hydrolyzate, sugar, adjuvants, a photoactive dye toxicant, and other additives was applied to the sponge. This station, when tested in an orchard, reduced sterile released adult populations by 70-90% in 4 d compared with check plots. Other tests in field cages showed that the bait station was approximately 22% less effective in killing adults compared with spot sprays on trees. We formulated a gelled bait by using a more refined hydrolyzed protein, supplemental attractants, feeding stimulants, and additives to protect the bait from drying. A series of experiments were carried out in field cages by using a cylindrical bait station that provided improved protection of the bait. These tests showed that there is a gradual decline in bait effectiveness with age. PMID- 17461070 TI - Knockdown, residual, and antifeedant activity of pyrethroids and home landscape bioinsecticides against Japanese beetles (Coleoptera: Scarabaeidae) on Linden foliage. AB - Residual toxicity and leaf protection capability of five pyrethroids, professional and home garden azadirachtin formulations, and six other bioinsecticides for the home landscape were evaluated against the Japanese beetle, Popillia japonica Newman (Coleoptera: Scarabaeidae), on linden, Tilia cordata L. Capacity of intoxicated beetles to recover and subsequently feed and disperse also was evaluated to provide insight on activity characteristics of the different compounds. Intact shoots were sprayed and left in the field for varying intervals before being challenged with beetles in no-choice and choice tests. All pyrethroids except permethrin gave greater leaf protection, knockdown, and kill than did carbaryl, the standard, after 14 d of weathering. Deltamethrin, cyfluthrin, bifenthrin, and lamda-cyhalothrin gave a high level of protection for at least 19 d, and azadirachtin (Azatin XL) deterred feeding in choice tests for as long as 14 d. Home garden formulations containing pyrethrins in canola oil (Pyola) or azadiractin (Neem-Away) gave good short-term (< 3-d) protection. Formulations of capsaicin, rotenone + pyrethrins, kaolin particle film, D limonene, or garlic extract were ineffective, the latter two formulations being highly phytotoxic to linden. Results of this study should help support updating of guidelines for insecticidal control of Japanese beetles. PMID- 17461071 TI - Activity of an essential oil derived from Chenopodium ambrosioides on greenhouse insect pests. AB - This study involved both greenhouse and laboratory experiments evaluating the effect of an essential oil product (QRD 400) derived from Chenopodium ambrosioides variety nr. Ambrosioides L. (Chenopodiaceae) on greenhouse insect pests that feed on different plant parts: citrus mealybug, Planococcus citri (Risso); longtailed mealybug, Pseudococcus longispinus (Targioni Tozzetti); western flower thrips, Frankliniella occidentalis (Pergande), and fungus gnats (Bradysia spp.). Treatments were applied to coleus, Solenostemon scutellarioides plants; transvaal daisy, Gerbera jamesonii flowers; or growing medium, depending on the insect pest. The essential oil was most effective, based on adult emergence, on both the second and third instars of the fungus gnat Bradysia sp. nr. coprophila when applied as a drench to growing medium. In addition, there was a significant rate response for QRD 400 on fungus gnats. The QRD 400 treatment had the highest percentage of mortality on longtailed mealybug (55%) compared with the other treatments. However, the essential oil was less effective against citrus mealybug (3% mortality) and western flower thrips adults (18-34% mortality) compared with standard insecticides, such as acetamiprid (TriStar) and spinosad (Conserve), which are typically used by greenhouse producers. This lack of efficacy may be associated with volatility and short residual properties of the essential oil or with the essential oil taking longer to kill insect pests. Other insecticides and miticides evaluated, including sesame oil, garlic, paraffinic oil, and Bacillus thuringiensis subsp. israelensis, provided minimal control of the designated insect pests. In addition, adult rove beetle Atheta coriaria Kraatz adults were not effective in controlling the larval instars of fungus gnats when applied at a rate of five adults per container. PMID- 17461072 TI - Genetic relationship between Coptotermes gestroi and Coptotermes vastator (Isoptera: Rhinotermitidae). AB - The phylogenetic relationship of Coptotermes gestroi (Wasmann) and Coptotermes vastator Light (Isoptera: Rhinotermitidae) was determined using DNA sequence comparisons of mitochondrial genes. Partial sequences of the ribosomal RNA small subunit 12S, ribosomal RNA large subunit 16S, and mitochondrial COII were obtained from nine populations of C. gestroi from South East Asia (Malaysia, Singapore, Thailand, and Indonesia) and four populations of C. vastator from the Philippines and Hawaii. In addition, four populations of Coptotermes formosanus Shiraki and Globitermes sulphureus (Haviland) were used as the outgroups. Consensus sequences were obtained and aligned. C. vastator and C. gestroi are synonymous, based on high sequence homology across the 12S, 16S, and COII genes. The interspecific pairwise sequence divergence, based on Kimura 2-parameter model between C. gestroi and C. vastator, varied only up to 0.80%. Morphometric measurements of 16 characteristics revealed numerous overlaps between the examined individuals of both species. Based on the molecular phylogenetics and morphometric data, it is proposed that C. vastator is a junior synonym of C. gestroi. PMID- 17461073 TI - Hermetically sealed baits for subterranean termites (Isoptera: Rhinotermitidae). AB - Cellulose baits containing 0.5% hexaflumuron were hermetically sealed in a closed cell polyethylene sheet envelope and placed in soil to test their durability and efficacy against field colonies of the Formosan subterranean termite, Coptotermes formosanus Shiraki, or the eastern subterranean termite, Reticulitermes flavipes (Kollar). The closed cell polyethylene sheet was readily tunneled through by termites, yet it was impervious to water and protected the cellulose baits and hexaflumuron from the environment. Only in a few incidents did the polyethylene envelope become infiltrated by plant roots, resulting in water intrusion and apparent degradation of cellulose baits. After consuming one to three sealed baits, three colonies each of both termite species were eliminated. The sealed baits may be placed in soil for months or years without the need of monitoring, and they are readily penetrated and fed upon by termites when they are present. Application of baits hermetically sealed in a protective sheet may save labor costs by bypassing the monitoring phase, circumvent the station avoidance by some termite species, and enable the use of baiting technologies in large areas such as agricultural fields in which the manual monitoring is impractical. PMID- 17461074 TI - Flybrella: a device to attract and kill house flies. AB - Flybrella is a lightweight inexpensive trap that can be suspended like an upside down umbrella in prominent locations where house flies, Musca domestica L., rest. It consists of a solid, cylindrical backbone to which is attached a perforated transparent tube, or baffle, with a commercial sugar/toxicant strip affixed inside. Centered directly beneath the tube and also attached to the backbone is a 10-cm-diameter inverted opaque plastic cone. House flies readily enter the tube, feed on the fast-acting toxicant, and then fall directly down the tube where they are collected in and concealed by the cone. The cone may be emptied easily through a capped opening in the bottom. In paired indoor tests, the efficacy of the commercial sugar/toxicant strip was increased significantly by the individual addition of several other attractant materials. Variants of the original design were tested, including a more efficient design featuring two concentric tubes with offset perforations. The toxicant strip may be easily removed and/or replaced when desired. PMID- 17461075 TI - Historical review of termite activity at forest service termiticide test sites from 1971 to 2004. AB - The U.S. Forest Service has a long history of providing termiticide efficacy data used for product registration and labeling. Four primary test sites (Arizona and Florida, Mississippi, and South Carolina [hereafter southeast]) have been used for this purpose. Various parameters of termite attack at water-only control plots were examined in this study to assess the relative pressures of termites at each site. Termiticide studies installed between 1971 and 2001 by using ground board (GB) and concrete slab (CS) test methods were included. GB control plots were attacked 85% of the time in the southeast, about twice the rate observed in Arizona (43%). CS plots were attacked 59-70% of the time in the southeast, significantly higher than in Arizona (43%). Termites were slower to initiate attack at control plots in Arizona compared with the southeast, and they were up to twice as slow at GB controls. Once initial attack began, GB plots were reattacked at higher percentages in the southeast (89-90%) than in Arizona (67%). Reattack at CS plots ranged from 65% in Arizona and South Carolina to 76% in Mississippi. Termites caused less damage to wooden blocks in control plots in Arizona than the southeast. Attack rates at controls generally declined during the 1990s, but these rates have rebounded since 2000, except at CS plots in Arizona and South Carolina. Statistical analysis of attacks at plots treated with chlorpyrifos, cypermethrin, fenvalerate, and permethrin also was undertaken. Time to initial termite attack (failure) of the organophosphate chlorpyrifos was generally shorter in Arizona than in the southeast, whereas time to initial attack in plots treated with one of three pyrethroids (cypermethrin, fenvalerate, and permethrin) was generally longer in Arizona. PMID- 17461076 TI - Toxicity, uptake, and transfer efficiency of fipronil in western subterranean termite (Isoptera: Rhinotermitidae). AB - The potential horizontal transfer ofnonrepellent termiticides has become an important paradigm to control termites in recent years. In this study, 14C radiolabeled fipronil was used in a series of laboratory experiments to demonstrate the extent and ability of termites to transfer lethal amounts of fipronil to unexposed nestmates. Fipronil is an active and nonrepellent termiticide against western subterranean termites, Reticulitermes hesperus Banks, on sand at relevant doses. It exhibited delayed toxicity with the lowest LD50 approximately 0.2 ng/termite expressed between day 4 and 7. Both continuous and brief exposures to fipronil-treated sand seriously impaired the termite's ability to move and respond to a dodecatrienol trail, limiting potential horizontal transfer. In tunneling studies, fipronil prevented termite tunneling at concentrations as low as 0.5 ppm and was nonrepellent even at 500 ppm. Greater than 90% mortality was recorded by day 7 with concentrations ranging from 0.5 to 500 ppm. There was a linear relationship between the time of exposure and uptake of [14C]fipronil when termites were continuously exposed to 0.5, 1.0, and 5.0 ppm for 24 h. However, uptake discontinued when the termites were immobilized. Maximum transfer of fipronil from donors to recipients occurred within the first 24 h. Fipronil was transferred by body contact and trophallaxis did not play a major role in horizontal transfer. In successive transfer studies, there was not enough fipronil on recipients for them to serve as secondary donors and kill other termites. In a linear arena study, there was an inverse relationship between the amount offipronil on dead termites and their distance from the treated zone. Maximum mortality was observed within 1.5 m from the treated zone. Results in our laboratory studies suggest that horizontal transfer was not a major factor contributing to the efficacy of fipronil in the field. PMID- 17461077 TI - Relative attractiveness of baits to Paratrechina longicornis (Hymenoptera: Formicidae). AB - Exotic ant incursions are becoming more frequent around the globe, and management with toxic baits is a suitable strategy for most species. Crazy ants, (Latreille) (Hymenoptera: Formicidae), however, are notoriously difficult to attract to commercial baits, which are generally tailored to the preferences of fire ants. We tested P. longicornis preferences for various food types and commercial ant baits. Baits trialed were commercially available products Amdro, Maxforce, Xstinguish (nontoxic monitoring version), Presto, and tuna (in spring water), sugar water (25%), boric acid (1% in 25% sugar water), and deionized water. Tuna and Xstinguish, along with sugar water and sugar water + boric acid, were the most attractive baits to P. longicornis foragers. The granular baits (Maxforce, Amdro, and Presto) were not as attractive to P. longicornis foragers. A decrease in temperature from summer (30 degrees C) to autumn (23 degrees C) trials did not seem to affect the food preferences of P. longicornis. Although P. longicornis recruitment was substantially lower during trials where there was concurrent high native ant abundance and diversity, P. longicornis still recruited to preferred baits in numbers higher than any other species. Given that tuna is impractical for management programs, the effectiveness of boric acid, sweet liquid baits in eliminating P. longicornis colonies should be compared with that of the toxic version of Xstinguish. If both are effective at eliminating colonies, we recommend sweet liquid baits containing boric acid be used for small-scale incursions (one or two nests), but a more practicable solid bait, such as Xstinguish, be used for larger scale incursions (numerous nests). PMID- 17461078 TI - Landscape mulches and termite nutritional ecology: growth and survival of incipient colonies of Coptotermes formosanus (Isoptera: Rhinotermitidae). AB - Alate swarms are one of the major visible signs of the expansion of the Formosan subterranean termite, Coptotermes formosanus Shiraki (Isoptera: Rhinotermitidae), in an area. Successful establishment of an incipient colony is thought to mainly rely on available food resources and moisture. The large-scale use of tree-based mulches in landscapes may inadvertently contribute to local establishment and growth of C. formosanus colonies. This research investigated the nutritional ecology of incipient colonies of C. formosanus feeding on seven tree-based, weathered, and nonweathered landscape mulches: pine straw, pine bark, cedar wood, water oak, eucalyptus, cypress, and melaleuca. Incipient colonies of C. formosanus feeding on pine straw, either weathered or nonweathered, produced significantly more progeny over the course of 1-yr feeding than colonies feeding on the other mulches tested. Regardless of weathered or not, the incipient colonies feeding on pine straw, eucalyptus, bald cypress, and water oak mulches had significantly greater survival rates after 360 d (53-77%) than colonies feeding on the other mulches tested (0-13%), but colonies feeding on nonweathered water oak had significantly lower survival (8%) than those kept on weathered water oak (58%). Colony fitness values were significantly different between the weathering treatment groups and among the different types of mulches. With regard to colony growth characteristics, three distinct growth patterns were identified: a high number of progeny (>100) with high colony survival rate (>50%), a medium number of progeny (12-50) with high colony survival rate (>50%), and a small number of progeny (0-10) with low colony survival rate (<5%). These findings suggest that different types of mulch substrates could significantly impact the nutritional ecology of the founding pairs and the successful establishment of incipient colonies during the swarming season. PMID- 17461079 TI - Indirect selection for increased susceptibility to permethrin in diamondback moth (Lepidoptera: Plutellidae). AB - We have been exploring the behavioral response ot insect pests to heterogeneous distribution of toxins (low dose with refugia), and its genetic correlation with physiological tolerance to these toxins. A field-collected population of diamondback moth, Plutella xylostellu (L.) (Lepidoptera Plutellidae), from Celeryville, OH, was selected with permethrin to determine whether low heterogeneous doses could lead to increased susceptibility to permethrin by selecting indirectly on behavior. Two replicates of each of three selection regimes: uniform high concentration hypothesized to result in increased physiological tolerance, heterogeneous low concentration hypothesized to result in increased susceptibility through indirect selection on behavior, and a control with no exposure to permethrin, were maintained in 1-in3 cages in a greenhouse, for 33 generations. All life stages of the diamondback moth were exposed to the selection regimes, and new generations were started with a random selection of pupae from the previous generation. Lines selected with uniform high concentrations developed 76-fold levels of resistance to permethrin by the 17th generation, with little changes thereafter. For generations 1-20, lines selected with heterogeneous low concentrations remained slightly lower in LC50 but not significantly different from the unselected control lines. Based on confidence intervals from probit analyses, the LC50 of the lines selected with heterogeneous low concentration, however, were significantly lower than those of the control lines in generations 21-33. Our results demonstrate that selection on behavioral responses can result in greater susceptibility than no selection at all, despite exposure to the toxin and ample genetic variation and potential for increased physiological tolerance. The implications of our findings, which are based on selection scenarios that could take place in field situations, are that behavioral responses can prevent and even decrease the levels of resistance in insect populations, an important result with respect to resistance and resistance management. PMID- 17461080 TI - Toxicity, synergism, and neurological effects of novel volatile insecticides in insecticide-susceptible and -resistant Drosophila strains. AB - Naturally derived volatile insecticides from the heterobicyclic and formate ester classes were investigated using a combination of volatility and synergist bioassays. In these studies, Drosophila melanogaster (Meigen) was used as a model for other medically important dipterans. In addition to a susceptible strain (Canton-S), three mutant strains were tested that included a strain resistant by P450-based metabolism (Hikone-R) and two resistant neurological mutant strains; one voltage-gated sodium channel mutant (para(ts-1)) and one GABA-gated chloride channel mutant (Rdl). In general, the 11 tested insecticides displayed a diversity of toxicity, metabolism, and resistance characteristics that correlate with their structural diversity. Several important trends were revealed by these studies, including hydrolase- and cytochrome P450 (P450)-based activation, P450 based resistance, distinct patterns of neurological activity, and negative cross resistance with established insecticides. These findings provide important insight into the metabolism and modes of action for the volatile insecticides. These findings also suggest potential approaches for insecticide deployment in integrated vector management and resistance management programs. PMID- 17461081 TI - Insecticide susceptible/resistance status in Aedes (Stegomyia) aegypti and Aedes (Stegomyia) albopictus (Diptera: Culicidae) in Thailand during 2003-2005. AB - Susceptibility baselines and diagnostic doses of the technical grade insecticides deltamethrin, permethrin, fenitrothion, and propoxur were established based on Aedes aegypti (L.), Bora (French Polynesia), a reference susceptible strain. Field-collected Aedes mosquitoes from each part of Thailand were subjected to bioassay for their susceptibility to the diagnostic doses of each insecticide. Almost all Ae. aegypti collected were incipient resistant or resistant to deltamethrin and permethrin, except those from some areas of Songkhla (southern) and Phan district of Chiang Rai (northern) province. Susceptibility to fenitrothion was found in mosquitoes from Bangkok (central), Chonburi (eastern), Chiang Rai, Kanchanaburi (western), and Songkhla, whereas they were resistant in almost all areas of Nakhon Sawan (north central) and Nakhon Ratchasima (northeastern) provinces. Most of Ae. aegypti were susceptible to propoxur except those from Mae Wong, Nakhon Sawan province. Various levels of insecticide resistance and susceptibility in adjacent areas revealed a focal susceptible/resistance profile in the country. It could be noted that almost all of Ae. albopictus were susceptible to the insecticides tested at the same diagnostic doses. In conclusion, resistance to pyrethroids (permethrin and deltamethrin) has developed in Ae. aegypti in most of the collected areas, suggesting that an alternative choice of insecticide or other control measures should be applied. PMID- 17461082 TI - Evaluation of azinphos-methyl resistance and activity of detoxifying enzymes in codling moth (Lepidoptera: Tortricidae) from central Chile. AB - Regular applications of insecticides have been the main management practice against codling moth, Cydia pomonella (L.) (Lepidoptera: Tortricidae) in Chile. Organophosphates are the most widely used insecticides, and azinphos-methyl is an important element in spray programs. In particular, we evaluated diagnostic doses of azinphos-methyl on neonate and postdiapausing larvae from seven apple (Malus spp.) orchards. We also evaluated the activity of detoxifying enzymes, such as glutathione S-transferases (GSTs), cytochrome P450 polysubstrate monooxygenases (PSMOs), and esterases, which are likely to be involved in resistance to insecticides. Such responses were compared with an insecticide-susceptible strain that has been maintained in the laboratory for several years. Neonate larval mortality of field populations to azinphos-methyl was not significantly different from of the susceptible strain. In contrast, postdiapause larval mortality was significantly lower in the six analyzed populations than in the susceptible strain. The C. pomonella populations with reduced postdiapause mortality to azinphos-methyl also showed statistically higher GST activity. Finally, no significant differences were found in total esterase or PSMO activity between C. pomonella populations. Therefore, the observed reduction in postdiapause larval mortality to azinphos-methyl seems to be associated with an increase in GST activity. PMID- 17461083 TI - Interactions of alternate hosts, postemergence grass control, and rootworm resistant transgenic corn on western corn rootworm (Coleoptera: Chrysomelidae) damage and adult emergence. AB - Field studies were conducted in 2003 and 2004 to determine the effects of grassy weeds, postemergence grass control, transgenic rootworm-resistant corn, Zea mays L., expressing the Cry3Bb1 endotoxin and glyphosate herbicide tolerance (Bt corn), and the interactions of these factors on western corn rootworm, Diabrotica virgifera virgifera LeConte, damage and adult emergence. Three insect management tactics (Bt corn, its nontransgenic isoline, and isoline plus tefluthrin) were evaluated with two weed species (giant foxtail, Setaria faberi Herrm, and large crabgrass, Digitaria sanquinalis L. Scop), and four weed management regimes (weed free, no weed management, early [V3-4] weed management and late [V5-6] weed management) in a factorial arrangement of a randomized split split-plot design. In each case, the isoline was also tolerant to glyphosate. Root damage was significantly affected by insect management tactics in both years, but weed species and weed management did not significantly affect damage to Bt corn in either year. Adult emergence was significantly affected by insect management tactics in both years and by weed species in 2003, but weed management and the interaction of all three factors was not significant in either year. The sex ratio of female beetles produced on Bt corn without weeds was generally greater than when weeds were present and this difference was significant for several treatments each year. Average dry weight of male and female beetles emerging from Bt corn was greater than the weights of beetles emerging from isoline or isoline plus tefluthrin in 2003, but there was no difference for females in 2004 and males weighed significantly less than other treatments in 2004. The implications of these results in insect resistance management are discussed. PMID- 17461084 TI - Resistance of sweetpotato genotypes to adult Diabrotica beetles. AB - Production of sweetpotatoes, Ipomoea batatas (L.) Lam. (Convolvulaceae), is limited by several insect pests, including Diabrotica spp. (Coleoptera: Chrysomelidae), and new integrated pest management (IPM) techniques for this crop are needed. Host plant resistance is one attractive approach that fits well into IPM programs. A host plant resistance research program typically depends on reliable bioassay procedures to streamline evaluation of germplasm. Thus, a bioassay technique was developed for evaluating sweetpotato germplasm by using adults of the banded cucumber beetle, Diabrotica balteata LeConte, and spotted cucumber beetle, Diabrotica undecimpunctata howardi Barber. A single beetle was placed on a piece of sweetpotato peel (periderm and cortex with stele removed) that was embedded periderm-side up in plaster in a petri dish. Feeding and longevity of insects on 30 sweetpotato genotypes were evaluated in two experiments by using this procedure. Adult longevity ranged from 7 to 11 d for starved individuals to 211 d for beetles fed a dry artificial diet. Longevity of banded cucumber beetles that fed on sweetpotato peels ranged from 12 d for the most-resistant genotype to 123 d for SC1149-19, a susceptible control cultivar. Longevity of spotted cucumber beetles was slightly shorter than longevity of banded cucumber beetles. For the most resistant sweetpotato genotypes, both Diabrotica species exhibited a significant delay in initiation of feeding, and more beetles died on these genotypes before they had fed. Both antibiosis and nonpreference (antixenosis) are important mechanisms of resistance in sweetpotato genotypes. This bioassay was consistent with field results, indicating that this technique could be useful for evaluating resistance to Diabrotica spp. in sweetpotato genotypes. PMID- 17461086 TI - Quantification of ammonia release from fruit fly (Diptera: Tephritidae) attractants using infrared spectroscopy. AB - Ammonia is the primary attractant for tephritid fruit flies, and traps baited with synthetic attractants using ammonia formulations have been highly successful in capturing these pests. However, difficulties in quantifying release rates of ammonia have limited abilities to make comparisons among field tests of different species by using different formulations. Therefore, Fourier transform infrared (FTIR) spectroscopy was evaluated as a method to quantify ammonia from synthetic lures. Analysis of the headspace from commercial ammonium bicarbonate and ammonium acetate lures indicated that there is a large burst of ammonia liberated upon initial exposure of the lures, but after 5-7 d the release rates stabilize and remain steady for at least 60 d under laboratory conditions. During the period of steady release, FTIR st atic measurements showed a n average of 0.12 and 0.21 microg of ammonia per 50-ml sample from ammonium bicarbonate and ammonium acetate lures, respectively. FTIR dynamic measurements from ammonium acetate lures indicated a steady release rate of approximately 200 microg/h. Ammonia release rate from ammonium acetate lures could be reduced by decreasing the surface area of the release membrane, and the presence of crystal formations on the membrane seemed to decrease the longevity of the ammonium acetate lures. PMID- 17461085 TI - Evaluation of natural and engineered resistance mechanisms in potato against Colorado potato beetle in a no-choice field study. AB - The Colorado potato beetle, Leptinotarsa decemlineata Say, is the major insect pest of potato, Solanum tuberosum L., in eastern North America and is renowned for resistance development, currently resistant to >40 insecticides worldwide. Host plant resistance may assist in delaying in resistance development to insecticides. We evaluated natural host plant resistance mechanisms (glandular trichomes and Solanum chacoense Bitter-derived resistance) and engineered resistance mechanisms (Bacillus thuringiensis [Bt] Berliner cry3A and cry1Ia1) in a no-choice cage study. Six different potato lines representing four host plant resistance mechanisms were evaluated over 2 yr. Egg masses were placed in each cage (one egg mass per plant). Almost no feeding was observed in the Bt-cry3A lines, and only minor feeding was observed in the Bt-cry1Ia1 lines in either year. On the S. chacoense-derived line, there was significantly less defoliation than on either the susceptible line or the glandular trichome line in 2003. In 2004, there was significantly higher defoliation on the S. chacoense-derived line than on the susceptible line or glandular trichome line. The defoliation of the Solanum chacoense-derived line was largely due to larvae clipping the petioles, rather than consumption of the leaves. Defoliation on the glandular trichome line did not differ significantly from the defoliation of the susceptible line, suggesting glandular trichomes may not be effective in controlling larvae and preventing defoliation. This study suggested that Bt can provide high levels of resistance, but the natural resistance mechanisms tested here are variable for control of Colorado potato beetle larvae in no-choice situations. PMID- 17461087 TI - Suppressive potential of bean (Phaseolus vulgaris) flour against five species of stored-product mites (Acari: Acarididae). AB - Previous research has demonstrated that legume proteins have insecticidal activity against stored-product pests, but activity against stored-product mites has not been tested. A study was therefore conducted to explore the potential of bean, Phaseolus vulgaris L., flour as novel botanical acaricide against five species of storage and dust mites: Acarus siro L., Aleuroglyphus ovatus (Troupeau), Caloglyphus redickorzevi (Zachvatkin), Lepidoglyphus destructor (Schrank), and Tyrophagus putrescentiae (Schrank). The effect of wheat, Triticum aestivum L., grain enriched with bean flour to eight concentrations (0, 0.01, 0.1, 0.5, 1, 2.5, 5, and 10%) on population growth initiating from the density of 50 mites per 100 g of wheat was recorded for 21 d under laboratory conditions (grain moisture 14.6% moisture content and 25 degree C in darkness). The enrichment of grain with bean flour suppressed the population growth of all tested species: 0.01% concentration reduced population growth of all tested species to >50% in comparison with the control population. The most sensitive species were A. siro and L. destructor, followed by T. putrescentiae and C. redickorzevi. The least sensitive species was A. ovatus. The terminal (i.e., after 21 d) density of mites positively correlated with bean flour concentration. The suppressive effect of bean flour was not linear but rather asymptotic. The results of this study are discussed in the context of the application of bean flour in integrated control of stored-product mites and the elimination of stored product mite allergens. PMID- 17461088 TI - Host-foraging success of three species of Trichogramma (Hymenoptera: Trichogrammatidae) in a simulated retail environment. AB - Three species of trichogrammatid egg parasitoids (Trichogramma deion Pinto & Oatman, Trichogramma ostriniae Pang & Chen, and Trichogramma pretiosum Riley) (Hymenoptera: Trichogrammatidae) were evaluated under laboratory conditions as potential biological control agents for the Indianmeal moth, Plodia interpunctella (Hubner) (Lepidoptera: Pyralidae), on retail shelves. A single shelving unit was used in each trial and a grid of sentinel egg disks was used to evaluate foraging success. The shelving consisted of pallet units with five shelves that were either bare or stocked with empty cereal boxes. In each replicate, approximately 500 female Trichogramma were released at the center of the shelving unit and allowed to forage for 48 h. Percentage of egg parasitism and percentage of host egg mortality were recorded after 7 d. Foraging success as well as the spatial pattern of parasitism differed significantly among the three Trichogramma species. Percentage of egg parasitism was approximately 4 times greater for T. deion than for T. ostriniae or T. pretiosum. The vertical distribution of parasitism by T. deion was also more uniform than for the other two species. In addition, the presence of packaging affected the foraging efficiency of T. ostriniae and T. pretiosum but not T. deion. Based on these findings, Trichogramma deion may be the best-suited candidate for augmentative biological control of P. interpunctella in retail stores, and a central release point of T. deion will likely provide adequate coverage of products on pallet type shelving. PMID- 17461089 TI - Influence of temperature and humidity on insecticidal effect of three diatomaceous earth formulations against larger grain borer (Coleoptera: Bostrychidae). AB - Laboratory experiments were carried out to evaluated three diatomaceous earth (DE) formulations--Protect-It, PyriSec (at dose rates 500, 1000, and 1500 ppm), and DEA-P (at dose rates 75, 150, and 500 ppm)--against the larger grain borer, Prostephanus truncatus (Horn) (Coleoptera: Bostrychidae), adults in stored maize, Zea mays L., at three temperatures (20, 25, and 30 degrees C) and two relative humidity (RH) levels (55 and 75%). At these conditions, the capability of progeny production in the treated substrate also was assessed. Adult survival was high, at all doses of Protect-It and PyriSec. Progeny production was also high. In contrast with the other two DEs, DEA-P was highly effective and caused complete mortality to the exposed P. truncatus adults, even at the lowest dose rate (75 ppm). In addition, progeny production was completely suppressed. Generally, Protect-it and PyriSec were more effective at 20 degrees C than at 30 degrees C. In contrast, the efficacy of DEA-P was continuously high in all temperatures and relative humidities examined. PMID- 17461090 TI - Cost and risk analysis of heat and chemical treatments. AB - An economic evaluation of newly developed methods for disinfesting empty grain storage bins by heat treatment will be a useful tool for decision-making by grain storage managers. An economic empirical model of heat treatment and chemical applications was developed using minimization of costs at a target risk level associated with the grain-damaging insects Tribolium castaneum (Herbst), Sitophilus oryzae (L.), and Rhyzopertha dominica (F.). Risk was measured as a deviation below a target mortality goal (Target MOTAD). Insect mortality and air temperature during heat treatment were evaluated for empty storage bins with a full drying floor, along with a similar evaluation of insect mortality for two application rates of a contact pyrethroid insecticide, cyfluthrin 20% active ingredient (AI) wettable powder. A high-output propane heater (29 kW) had the lowest cost and risk level of all heating systems and produced 100% mortality in 2 h for the three insect species at all test locations. An electric duct-heater system (18 kW) also produced 100% mortality at all test locations after 40 h, but it had significantly higher costs. The other heating system configurations in the study had significantly higher risk levels of insect mortality, and the electric systems were not cost-effective. Both chemical rates had low costs and risk levels, with high mortality results. PMID- 17461091 TI - Performance of painted plywood and cloth Nzi traps relative to Manitoba and greenhead traps for tabanids and stable flies. AB - Experiments were conducted to adapt the cloth Nzi trap to a format suitable for fixed applications in biting fly sampling or control. Catches of tabanids [Tabanus L., Chrysops (Meigen), and Hybomitra Enderlein], and stable flies [Stomoxys calcitrans (L.)] in painted plywood traps were compared with those in standard phthalogen blue cloth traps, and in similarly painted cloth traps. The Manitoba horse fly trap and the Tabanus nigrovittatus Macquart "greenhead" box trap were used as additional standards during one tabanid season. Shiny features of traps reduced catches, e.g., paint on cloth instead of wood, or use of aluminum screening instead of netting. Nevertheless, appropriately painted plywood Nzi traps caught as many biting flies as did standard cloth Nzi traps, if paint finishes were matte, and with the use of phthalogen blue colorants. Nzi traps collected about the same tabanid fauna as the Manitoba and T. nigrovittatus traps, but with improved catches of Chrysops and Tabanus. Recommendations are provided on appropriate color matching, and selection of readily-available materials for trap construction. PMID- 17461092 TI - Life cycle details of Solenopotes capillatus (Anoplura: Linognathidae). AB - The duration of immature life stages and the preoviposition period of Solenopotes capillatus Enderlein (Anoplura: Linognathidae) were determined by daily observation in vivo of individual specimens. The duration of the egg and nymphal stages was obtained by daily observation of eggs deposited by females transferred to normal predilection sites of uninfested cattle. The preoviposition period was determined by transferring third instars to uninfested cattle and then recording the period from adult eclosion to first oviposition. The egg stage required 11-13 d. Both first and second instars required 3-4 d, and third instars, based on observations of females, required 5-6 d. The preoviposition period was 2 d. Between 27 and 29 d was required for completion of the cycle (egg to egg). The size (length) of unmounted specimens, in millimeters, was egg 0.70; first instar 0.69; second instar, 0.82; third instar, 1.06; adult female, 1.50; and adult male, 1.08. PMID- 17461093 TI - Biocidal activity of three wood essential oils against Ixodes scapularis (Acari: Ixodidae), Xenopsylla cheopis (Siphonaptera: Pulicidae), and Aedes aegypti (Diptera: Culicidae). AB - The biocidal activity of three steam distilled wood essential oils-incense cedar, Calocedrus decurrens (Torr.) Florin; Port-Orford-cedar, Chamaecyparis lawsoniana (A. Murr.) Parl.; and western juniper, Juniperus occidentalis (Hook)--were evaluated against adult Aedes aegypti (L.) (Diptera: Culicidae) and Xenopsylla cheopis (Rothchild) (Siphonaptera: Pulicidae) and nymphal Ixodes scapularis Say (Acari: Ixodidae). In vitro laboratory bioassays were conducted to establish baseline dose-mortality data through 24 h. Incense cedar heartwood was the most toxic to all three vector species followed in order of activity by western juniper and Port-Orford-cedar based on LC50 and LC90 values. Ae. aegypti were substantially more susceptible to the oils than either I. scapularis or X. cheopis. PMID- 17461094 TI - [Prebiotics and probiotics in infant nutrition]. AB - Many studies have recently analyzed the modulation of the intestinal microflora showing a benefic effects reducing the number of enteritis, improving the oligoelements absorption and stimulating the immunitary system. To do so three way are available: the use of prebiotics, the use of probiotics and the symbiotic way. Prebiotics are non-digestible oligosaccharides that can stimulate selectively the growth bifidogenus bacteria. Probiotics are dietary supplements made of live micro-organisms which improve the microbial environment of the gut. In this review literature is examined the possible efficacy of prebiotics and probiotics in the pediatric age; however, the studies available do not permit to obtain definitive conclusions. PMID- 17461095 TI - [Intensive care at extremely low gestational age: ethical issues and treatment choices]. AB - With the continuing progress of obstetrical and neonatal care, the limit of human viability has continued to shift towards younger gestational ages. The survival rate as well as the survival without disability increases with each additional week of gestation but, for infants born from 22 to 25 w GA, it is still really low, and the threshold of human viability appears to be limited to the physiological development of the lungs, which take place around the 23rd-24th w GA. At present, the care of such infants, born at the threshold of human viability, presents a variety of complex medical, social, and economical decisions assuming the characteristics of ethical decisions in that the limits between benefits and disadvantages is not so clearcut. What is the true infant's best interest is far to be understood and concern about the ethical basis of providing such intensive care is arising in the scientific community. In this paper the authors provide a review of the ethical basis of decisions related to the care of such infants: to treat, not to treat, intensive or compassionate care, withhold or withdraw treatment. PMID- 17461096 TI - [Association among postural and skull-cervico-mandibular disorders in childhood and adolescence. Analysis of 428 subjects]. AB - The aim of the study is to appraise the prevalence of the mild defects of the spine or paramorphysm in a normal paediatric population, without orthopaedic pathology, and to verify the possible association with the dysfunctions of the dental apparatus. The recovery of a real association among these pathologies could furnish a sprout for a correct therapeutic approach. In the study 428 subjects (211 females and 217 males), aged 9 to 14 years, have been analyzed. The data have been recorded on special schedules, orthopaedic and orthodontic, containing the number of progressive order and the initials of name and last name, in the respect of the privacy of the subjects. From the study a 2.8% scoliosis incidence has emerged, an incidence of scoliotic attitudes 9.5% and an incidence of 83% normality deviations. As deviations by the normality have been classified the isolated skeletal asymmetries, agreements as varying functional not yet pathological, for instance scapular or flank asymmetry, mild genu varum or valgum, femoral anteversion. A statistically significant relationship among that disorders of posture and malocclusion (P < 0.005) and ogival palate (P < 0.002) has been found. Despite these correlations, it is very difficult to explain this association from the point of view of etiopathogenesis. For this we hold to have to continue the study, to give a meaning to such correlations and to find an appropriate therapy. PMID- 17461097 TI - Early diagnosis and treatment of celiac disease in type 1 diabetes. A longitudinal, case-control study. AB - OBJECTIVES: Subjects with type 1 diabetes mellitus (DM) are at increased risk to develop celiac disease (CD). However, most of the published investigations on the association between type 1 DM and CD are cross-sectional ones. In this paper, longitudinal data are presented on the effects of gluten-free diet (GFD) on growth and metabolic control in children and adolescents with type 1 DM screened for CD. METHODS: Clinical records of 27 patients with type 1 DM+CD (7% of a population of 385 subjects with type 1 DM) were reviewed. The following variables were considered at the diagnosis of CD (T0) and after 24 (T24) and 48 (T48) months of GFD according to the length of available follow-up: weight, height, body mass index (BMI), Hb and HbA1c levels and per kg/day dose of insulin. Forty three patients with type 1 DM alone, matched for sex, age and duration of diabetes were chosen as controls. RESULTS: In patients with type 1 DM+CD, mean (SD) age at type 1 DM diagnosis was 8 (3.3) years. Median time interval between diagnosis of type 1 DM and of CD was 1.8 years (range 0.1-23.9 years). At T0 (n = 23), height and BMI z-score, HbA1c levels and daily insulin dose were comparable in type 1 DM+CD and in control subjects. Mean Hb concentration was significantly lower in subjects with type 1 DM+CD (12.6 (1.3) vs. 13.3 (0.7) g/dl, p < 0.05). At T24 (n = 22) and T48 (n = 16), no difference was detectable in height, BMI, HbAlc and insulin dose and also Hb concentration was comparable in both groups. CONCLUSIONS: This study shows that, at diagnosis of CD, screened subjects with type 1 DM had only minor signs of malnutrition and metabolic disturbances were uncommon. Dietary treatment of CD can allow a growth and diabetes control comparable with subjects with diabetes alone. PMID- 17461098 TI - [Two children with hip pain and fever]. AB - Myositis of the Obturator internus Muscle is an uncommon disease that usually strikes children and adolescents. It is characterized by fever, hip pain and limping. The more frequently etiologic agent involved is Stafylococcus aureus. In the diagnosis, the aid of imaging diagnostic techniques is critical. We report two adolescents with this condition. PMID- 17461099 TI - One trocar assisted pyeloplasty (OTAP): initial experience and codification of a technique. AB - BACKGROUND: The Anderson-Hynes pyeloplasty is still the gold standard for the surgical treatment of the ureteropelvic junction obstruction (UPJO) in children. To reduce the surgical invasivity, laparoscopic (RPSc) and retroperitoneoscopic (RPSc) pyeloplasty have been proposed but concerns have been expressed. In the RPSc approach, the small operative space and the technical difficulties have limited the application and diffusion of this techinique while in case of laparoscopy, a transperitoneal technique is used to treat a retroperitoneal disease. We report our initial experience with the "one trocar assisted pyeloplasty" (OTAP) in the treatment of UPJO in children. METHODS: From January 2005 to December 2006 sixteen children (13 males--3 females) from 2 to 18 months of age (mean age 5.6 months), with impaired monolateral renal function underwent surgical treatment for UPJO. In 11 cases the obstruction was on the left side and in one case a crossing vessel was found. The renal pelvis was anteriorly reached using a 10 mm operative telescope via a flank 12 mm incision. The UPJ was exteriorized and a dismembered pyeloplasty performed also for the crossing vessel. In two cases a 6 Fr uretero-pyelo-nephrostomy catheter was used. RESULTS: The patients were discharged after four days without complications. The cosmetic results are excellent. In one case, a small perianastomotic leakage was seen. The ultrasonographic follow-up demonstrated the spontaneous resolution of the leakage. The post operative US follow-up showed the reduction of the dilatation in all cases. DISCUSSION: This is an initial experience and a longer follow-up is needed. However, in our opinion the OTAP could be considered as a safe and feasible alternative to the laparoscopic or retroperitoneoscopic pyeloplasty, especially in very small children and also in presence of crossing vessels. PMID- 17461100 TI - [Ethical discernment of a "tragic" situation]. PMID- 17461101 TI - HPLC retention behavior on hydride-based stationary phases. AB - Two stationary phases attached to a silica hydride surface, cholesterol and bidentate C18, are investigated with a number of pharmaceutically related compounds in order to illustrate the various retention mechanisms that are possible for these bonded materials. The test solutes range from hydrophilic to hydrophobic based on log P (octanol/water partition coefficient) and pKa values. The mobile phases consist of acidified (formic and perchloric acid) water/methanol or water/ACN mixtures. Of particular interest are the high organic content mobile phase compositions where the retention would increase if the bonded material was operating in the aqueous normal phase (ANP) mode. Plots of retention factor (k) versus mobile phase composition are used to elucidate the retention mechanism. A number of examples are presented where solutes are retained based on RP, ANP, or dual retention mechanisms. The silica hydride-based stationary phases can also retain compounds in the organic normal phase. PMID- 17461102 TI - The effect of the pore structure and zeta potential of porous polymer monoliths on separation performance in ion-exchange mode. AB - Most often, in bioseparations involving charged macromolecules, the chromatographic systems have low Reynolds and high Peclet numbers. For such systems, an expression is developed and presented in this work for evaluating the throughput in polymeric monoliths where ion-exchange adsorption occurs, as a function of (i) the pressure drop along the length of the monolith, (ii) the functional form and width of the throughpore-size distribution of the monolith, and (iii) the magnitude of the zeta potential on the surface of the throughpores of the monolith. Gaussian and log-normal throughpore-size distributions whose mean throughpore-size and standard deviation values are based on experimentally measured throughpore-size distribution data by mercury porosimetry employed on polymeric monoliths are used in this work, and their effect on the throughput relative to that obtained from a polymeric monolith having a uniform throughpore size distribution is studied for different values of the ratio of the standard deviation to the mean throughpore-size. The results indicate that relatively modest increases in the throughput, when compared with the throughput that could be achieved in a polymeric monolith having a uniform throughpore-size distribution, could be obtained in polymeric monoliths having disperse throughpore-size distributions, and the magnitude of the increase becomes larger when the disperse distribution is skewed to larger throughpore sizes. Furthermore, the results of this work indicate that, under certain conditions, relatively modest increases in the throughput of a charged analyte could also be achieved by altering the value of the zeta potential on the surface of the throughpores of the monolith. Due to the difficulties inherent in controlling the functional form and width of the throughpore-size distribution during the synthesis of polymeric monoliths, it would appear to be more practical to increase the value of the throughput of a charged analyte by altering the value of the zeta potential through prudent selection of the ion-exchange surface functional groups and fine-tuned with the pH of the mobile phase. Thus, for ion exchange chromatography systems, the zeta potential could be considered an important parameter for column designers and operators to manipulate, since its alteration could increase the through-put of a charged analyte in polymeric monoliths or in columns packed with charged particles. PMID- 17461103 TI - Combination of COFRADIC and high temperature-extended column length conventional liquid chromatography: a very efficient way to tackle complex protein samples, such as serum. AB - The previously reported COmbined FRActional DIagonal Chromatography (COFRA-DIC) methodology, in which a subset of peptides representative for their parent proteins are sorted, is particularly powerful for whole proteome analysis. This peptide-centric technology is built around diagonal chromatography, where peptide separations are crucial. This paper presents high efficiency peptide separations, in which four 250 x 2.1 mm, 5 microm Zorbax 300SB-C18 columns (total length 1 m) were coupled at operating temperatures of 60'C using a dedicated LC oven and conventional LC equipment. The high efficiency separations were combined with the COFRADIC procedure. This extremely powerful combination resulted, for the analysis of serum, in an increase in the uniquely identified peptide sequences by a factor of 2.6, compared to the COFRADIC procedure on a 25 cm column. This is a reflection of the increased peak capacity obtained on the 1 m column, which was calculated to be a factor 2.7 higher than on the 25 cm column. Besides more efficient sorting, less ion suppression was noticed. PMID- 17461104 TI - Determination of trans-resveratrol in wine by micro-HPLC with fluorescence detection. AB - A new method for the quantitative determination of trans-resveratrol in wine has been optimized and validated. The method is based on the direct injection of 500 nL of wine in an HPLC system fitted with an RP microcolumn of 10 cm x 0.32 mm and spectrofluorometric detection. The linear dynamic range of the method covers the normal range of occurrence of the analyte in wines and extends for two orders of magnitude with r(2)= 0.9994. Twenty-three wines have been analyzed with the proposed method, finding concentrations in the range reported by other authors. PMID- 17461105 TI - Coupled-column liquid chromatography combined with postcolumn photochemical derivatization and fluorescence detection for the determination of herbicides in groundwater. AB - This study examines the application of coupled-column LC-photochemically induced fluorimetry-fluorescence detection (LC-LC-PIF-FD), demonstrating its potential for the quantitative and selective detection of six herbicides, including propanil and the phenylureas monuron, monolinuron, chlorotoluron, diuron and neburon in groundwater samples. An AQUASIL C18 50 x 4.6 mm(2) id column coupled to an AQUASIL C18 150 x 4.6 mm(2) id column for analyte clean-up and determination were used, respectively. A simple SPE with Cl8 cartridges was carried out, yielding average recoveries between 80 and 112% (n = 6) with RSDs between 0.5 and 9%. The LODs ranged from 0.0083 to 0.0833 microg/L in the groundwater samples. PMID- 17461106 TI - Development of a validated HPLC method for the determination of four 1,4 benzodiazepines in human biological fluids. AB - A simple and sensitive HPLC method was developed and validated for the determination of four frequently prescribed 1,4-benzodiazepines: alprazolam (ALP), bromazepam (BRZ), diazepam (DZP), and flunitrazepam (FNZ). Separation was achieved on an Inertsil C8 analytical (250 mm x 4 mm, 5 microm) column, after selective extraction of benzodiazepine drugs from biological matrices by means of SPE. Isocratic elution was performed with a mobile phase consisting of CH3COONH4, 0.05 M CH3OH, and CH3CN (33:57:10 by volume). Quantification was performed at 240 nm with mefenamic acid (6 ng/microL) as the internal standard. DSC-18 Supelco cartridges provided high absolute recoveries (81-115%). The developed method was fully validated in terms of selectivity, linearity, accuracy, precision, stability, and sensitivity. Repeatability (n = 8) and between-day precision (n = 8) revealed RSD <12%. Recoveries from biological samples ranged from 81.2 to 115%. The detection limit of the method was calculated as 3.3-10.2 ng in blood plasma and 2.6-12.6 ng in urine for 20 microL injection volume. The method was applied to spiked biological matrices. Moreover, the method was applied to real samples of urine after an oral administration. PMID- 17461107 TI - Kinetic modeling of proteins adsorption with a methodology of error analysis. AB - The estimation of adsorption parameters for chromatographic systems is a very important step for column characterization used in the design of continuous separation equipments. The turbulent hydrodynamics aspect of batch procedures makes the kinetic modeling an interesting tool for the process modeling used for the determination of the main parameters that will be considered in the equipment design. The implemented irreversible kinetic model, which depends on both solute and site concentrations, was found to be very effective in the experimental correlation of two different enzyme adsorption systems: adsorption of Inulinases and beta-Galactosidase using two different adsorbents, the CM-Sepharose CL-6B and Accell plus QMA, respectively. The implementation of an error analysis methodology associated with an inverse problem approach was successful in determining the kinetic parameters with high accuracy. The simulations indicated a decrease in the kinetic constants with an increase in the solute concentration, which can be related to the increase in the competition by sites of adsorption. PMID- 17461108 TI - Reduction of chloride matrix effect using silver oxide as a precipitating reagent for the determination of trace anions in chloride-rich samples via ion chromatography. AB - A new method was developed for the determination of trace anions in chloride-rich samples via ion chromatography by reducing the concentration of chloride ions using silver oxide as a precipitating reagent. In this method the sample pretreatment was started with the addition of silver oxide powder followed by 8 min of stirring and 2 min of centrifugation at a speed of 1500 rpm. The supernatant was diluted with a 1:10 ratio using deionized water and filtered with a 0.22 microm NC filter. All the samples were analyzed in a Dionex ICS-2500 system with an ED50 electrochemical detector in conductivity mode. On the basis of Dionex IonPac AG18 (50 mm x 2 mm) and AS18 (250 mm x 2 mm) columns, 25 AL sample filtrate was eluted isocratically with a mobile phase of 32 mmol/L NaOH at a flow rate of 0.25 mL/ min and each eluted analyte was detected after postcolumn suppression via a Dionex anion suppressor ULTRA II (2 mm). With this method most of the common anions at mg/kg level in chloride-rich samples can be accurately quantified. By controlling the equivalent weight ratio of silver oxide to chloride in sample solutions and the time of pretreatment, the free silver and chloride ions in the sample solution are optimized such that most coexisting common anions can be determined simultaneously. PMID- 17461109 TI - Comparison of the performance of different reversed-phase columns for liquid chromatography separation of 11 pollutant phenols. AB - A systematic optimization of the HPLC separation of a mixture containing 11 pollutant phenols (PPs) using a Hypersil ODS (250 mm x 4.6 mm, 5 microm) column and UV-DAD detection has been carried out. The binary mobile phases used were obtained by mixing 50 mM phosphate (pH = 3.0) and methanol, ACN, or THF as organic modifiers. After selecting ACN as an organic modifier, the effects of pH and temperature on PPs separation were studied. A mobile phase of 50 mM acetate (pH = 5.0)-ACN (60:40 v/v) at 50 degrees C allowed the separation of 11 phenols but not to baseline in 17 min. To improve the performance of this separation, the following RP columns were tested: Luna C18 (2), Purospher C18, Synergi C12, Synergi Fusion C18, Gemini C18, Luna Cyano, Lichrospher C8, and Envirosep-PP (polymeric). In all the cases, the performance (analysis time, retention, selectivity, resolution, asymmetry factors, and efficiency) was evaluated. A further reoptimization of the mobile phase was carried out for all the columns by studying the ACN content and pH, with the aim of improving the above-mentioned separations and selecting the most suitable one for PPs analysis. PMID- 17461110 TI - Comparison between HPLC and HPTLC densitometry for the determination of icariin from Epimedium koreanum extracts. AB - Dry extracts of the aerial parts of Epimedium koreanum were quantified by HPLC and high performance TLC (HPTLC). A gradient HPLC method was used for the quantification of the prenylflavone glycoside icariin at 270 nm. A direct HPTLC assay was developed for the determination of icariin at 270 nm. The UV detection of both analytical assays were used to examine the purity of icariin peaks and compared with the standards. The assays provide good accuracy, reproducibility, and selectivity for the quantitative analysis of icariin. The icariin contents of five different dry extracts were compared by HPLC and HPTLC densitometry. The quantitative results of both analytical methods did not show any statistically significant differences between them, although a trend to slightly lower mean values could be found for the HPLC method. PMID- 17461111 TI - Direct enantiomeric separations of tris(2-phenylpyridine) iridium (III) complexes on polysaccharide derivative-based chiral stationary phases. AB - A convenient method is presented for the first time for the direct separation of enantiomers of a tris(2-phenylpyridine) iridium (III) and an analog substituted with long alkoxy chains on polysaccharide derivative-based chiral stationary phases by HPLC. Tris(2-phenylpyridine) iridium (III) was separated on the immobilized amylose 3,5-dimethylphenylcarbamate (Chiralpak IA) using hexane/CHCl3/CH2Cl2 (75:20:5) as an eluent, and the analog could be separated on the coated cellulose 3,5-dimethyl-phenylcarbamate (Chiralcel OD) and cellulose 4 methylbenzoate (Chiralcel OJ) using hexane/2-propanol (96:4) as the eluent. CD spectra of the eluted HPLC fractions were also recorded, and the observed mirror image patterns confirm their enantioseparations. PMID- 17461112 TI - Simultaneous determination of vitexin-2"-O-glucoside, vitexin-2"-O-rhamnoside, rutin, and hyperoside in the extract of hawthorn (Crataegus pinnatifida Bge.) leaves by RP-HPLC with ultraviolet photodiode array detection. AB - RP-HPLC with UV photodiode array detection (UV-DAD) was developed and validated for the simultaneous determination of vitexin-2"-O-glucoside, vitexin-2"-O rhamnoside, rutin, and hyperoside in the extract of hawthorn (Crataegus pinnatifida Bge.) leaves. The analytes of interest were separated on a Diamonsil C18 column (250 x 4.6 mm id, 5 microm) with the mobile phase consisting of THF/ACN/methanol/ 0.05% phosphoric acid solution (pH 5.0) (18:1:1:80 v/vl/v). The flow rate was set at 1.0 mL/min and the eluent was detected at 340 nm for the four flavonoids. The method was linear over the studied range of 1.00-100 microg/mL for the four analytes of interest with the correlation coefficient for each analyte greater than 0.999. The LOD and LOQwere 0.03 and 0.10 microg/mL, 0.03 and 0.10 microg/mL, 0.05 and 0.15 pg/mL, 0.10 and 0.30 microg/mL for vitexin 2"-O-glucoside, vitexin-2"-0-rhamnoside, rutin, and hyperoside, respectively. The optimized method was successfully applied to the analysis of four important flavonoids in the extract of hawthorn leaves. The total amounts of the four flavonoids were 22.2, 62.3, 4.27, and 8.24 mg/g dry weight for vitexin-2"-O glucoside, vitexin-2"-O-rhamnoside, rutin, and hyperoside in the extract of hawthorn leaves, respectively. PMID- 17461113 TI - Assessment of the matrix effect on the headspace solid-phase microextraction (HS SPME) analysis of chlorophenols in wines. AB - In this paper, we propose a comparative study to check the matrix effect on the extraction of three chlorophenols, 2,4,6-trichlorophenol (TCP), 2,3,4,6 tetrachlorophenol and pentachlorophenol, direct precursors of 2,4,6 trichloroanisole, in synthetic and commercial wines (white and red wines). A rapid, simple and sensitive methodology based on solid-phase microextraction (SPME) and GC with electron capture detection (GC-ECD) and mass spectrometric detection (GC-MS) was developed and the variables affecting the extraction process (temperature, time and salt content) were examined employing a factorial design at two levels. Since GC-ECD does not allow the clear identification of target analytes in white wine, owing to overlapped interferences, GC-MS/MS was used for subsequent examinations. Calibration curves were constructed in synthetic, white and red wine. Significant differences between the slopes of synthetic and red wine, with the exception of TCP, were observed. Analytical parameters were evaluated and satisfactory results were obtained, showing the usefulness of the headspace SPME (HS-SPME) method for determining chlorophenolic compounds in wines. PMID- 17461114 TI - Characterization of substituted polyacetylene microstructure by pyrolysis gas chromatography. AB - A series of substituted acetylenes has been polymerized with WOC14/Ph4Sn metathesis catalyst and [Rh(cod)OMe]2 insertion catalyst, and the thermal degradation of the polyacetylenes prepared has been studied using pyrolysis capillary gas chromatography (Py-GC) with flame ionization and mass spectrometric detection to obtain information on the effect of the catalyst on the head-tail (H T) isomerism of polyacetylenes (poly(phenylacetylene), poly[(4 methylphenyl)acetylene], poly(benzylacetylene), poly((2-fluorophenyl)acetylene], poly[(3-fluorophenyl)acetylene], and poly[(4-fluoro-phenyl)acetylenel). Cyclotrimers have been found to be the main pyrolysis products in all cases. Direct Py-MS connection was used to determine the temperature profiles of the released pyrolysis products. 1,3,5-Trisubstituted benzenes were found to be the predominant pyrolysis products of the polymers prepared with the insertion catalyst, which proves the presence of long head-to-tail sequences of monomeric units in these polyacetylenes. On the other hand, both 1,2,4- and 1,3,5 trisubstituted benzenes are present in significant amounts in the pyrolysis products of polymers prepared with the metathesis catalyst, which proves the presence of a significant content of the head-to-head (HH) and tail-to-tail (TT) linkages in these isomers of polyacetylenes. Contents of the regular (HT) and inverse (HH-TT) monomer linkages (RML and IML, respectively) in polymer chains were determined from the relative amounts of di-, tri-, and tetrasubstituted benzenes found in the Py-GC products. PMID- 17461116 TI - Identification and determination of triterpenoids in Hieracium pilosella L. AB - Fractions of triterpenoids have been isolated from herb, inflorescences, and rhizomes with roots of Hieracium pilosella by typical extraction with petroleum ether. The fraction from inflorescences was investigated using GC-MS techniques. The occurrence of alpha- and beta-amyrin, taraxerol, taraxasterol, and fern-7en 3beta-ol has been observed. Quantitative analysis was also performed and taraxasterol is distinctly predominant in this triterpenoid fraction. Thin-layer chromatography on silica gel was performed on all the investigated fractions and additionally revealed the occurrence of lupeol and psi-taraxasterol in vestigial quantities. All of the triterpenoid components are reported for the first time in the investigated plant. PMID- 17461115 TI - Evaluation of the separation characteristics of application-specific (fatty acid methyl esters) open-tubular columns for gas chromatography. AB - The solvation parameter model is used to characterize the separation properties of the polar stationary phases EC-Wax and PAG with a poly(ethylene oxide) backbone (substituted with propylene oxide in the case of PAG) and the cyanopropyl-substituted polysilphenylene-siloxane stationary phase BPX90 at five equally spaced temperatures between 60 and 140 degrees C. The separation characteristics of these stationary phases are compared to four PEG and two poly(cyanopropylsiloxane) stationary phases (HP-20M, HP-Innowax, SolGel-Wax, DB WAXetr, HP-88, and SP-2340) characterized in the same way. The database of system constants for these polar stationary phases is used to provide insight into the separation mechanism for fatty acid methyl esters and to determine selectivity differences that can be expected for generically similar stationary phase types. The discussion is not structured to indicate which stationary phase should be used for a particular separation but to provide a general framework to demonstrate the relationship between the retention mechanism and stationary phase chemistry. PMID- 17461117 TI - Determination of aqueous inclusion complexation constants and stoichiometry of alkyl(methyl)-methylimidazolium-based ionic liquid cations and neutral cyclodextrins by affinity capillary electrophoresis. AB - Affinity CE (ACE) method was developed to characterize the complex formation between seven alkyl(methyl)methylimidazolium-based ionic liquid (IL) cations and eight neutral cyclodextrins (CD). The effective mobility data of the IL cations were processed according to classical nonlinear and linear treatments to obtain the complex stoichiometry and formation constant K. The majority of systems followed a 1:1 complexation stoichiometry model but in four cases a 1:2 stoichiometry was better satisfied. The K values obtained for each IL were compared to elucidate the main influences of IL and CD nature. The availability of these data should lend support to various application areas, including the screening and tailoring of new interactions in the solution for CE. PMID- 17461118 TI - Study of the complexation of different methacrylates with cyclodextrins employing a combination of electrophoretic, chromatographic, and NMR-spectroscopic methods. AB - The present study describes the application of capillary electromigration techniques; CEC and micellar EKC (MEKC), and the application of spectroscopic methods; 1H NMR and 1H NOESY spectroscopy to investigate interactions between CDs (alpha-CD, statistically methylated beta-CD, hydroxypropyl-beta-CD, and 2 hydroxypropyl-gamma-CD) and different methacrylates (adamantyl, isobornyl, cyclohexyl, and phenyl methacrylate). It is shown that these methods complement each other. While CD-mediated MEKC is a rapid screening technique for comparing complex stabilities in aqueous media, 1H NMR chemical shift analysis provides quantitative data for very strong methacrylate-CD complexes and CD-mediated CEC provides quantitative data for complexes with lower complex forming constants. CD mediated MEKC did not prove to be suitable for the calculation of complex forming constants. Reasons are discussed. 1H NOESY spectra were used to study spatial relationships between host and guest atoms. PMID- 17461119 TI - Quantitative determination of haloperidol in tablets by high performance thin layer chromatography. AB - A densitometric high performance thin-layer chromatography (HPTLC) method was developed and validated for the quantitative analysis of haloperidol in tablets. Chromatographic separation was achieved on precoated silica gel F 254 HPTLC plates using a mixture of acetone/chloroform/n-butanol/acetic acid glacial/water (5:10:10:2.5:2.5 v/v/v/v/v) as the mobile phase. Quantitative analysis was carried out at a wavelength of 254 nm. The method was linear in the 10-100 ng/microL range, with a determination coefficient of 0.999. The coefficients of variation for precision were not higher than 2.35%. The detection limit was 0.89 ng/microL, and the quantification limit was 2.71 ng/microL. The accuracy ranged from 97.76 to 100.33%, with a CV not higher than 4.50%. This method was successfully applied to quantify haloperidol in real pharmaceutical samples, including the comparison with HPLC measurements. The method was fast, specific, with a good precision and accuracy for the quantitative determination of haloperidol in tablets. PMID- 17461120 TI - [Historical profiles in stroke medicine]. PMID- 17461121 TI - [Neurosurgical treatment for stroke--past, present, future]. PMID- 17461122 TI - [Japanese Guidelines for Management of Stroke, 2004--present significance and future issues]. PMID- 17461123 TI - [Significance and development of Japanese Stroke Data Bank]. PMID- 17461124 TI - [Trends of stroke incidence and mortality in Japan and the world, and their prospects in near future]. PMID- 17461125 TI - [Trends in the type-specific incidence of stroke in a general Japanese population: the Hisayama Study]. PMID- 17461126 TI - [Current state and future perspective of management of acute ischemic stroke in Japan]. PMID- 17461127 TI - [Latest clinical trend of stroke epidemiology shown by Stroke Data Bank]. PMID- 17461128 TI - [Is the CVD-III classification of the NINDS still useful?]. PMID- 17461129 TI - [Characteristics and assessment of TOAST classification for diagnosing stroke subtypes]. PMID- 17461130 TI - [Cerebrovascular disorders and anatomy of the cerebrovascular system]. PMID- 17461131 TI - [Cerebral blood flow measurement in patients with stroke]. PMID- 17461132 TI - [Cerebrovascular disease and hemorheology]. PMID- 17461133 TI - [Neurochemistry and neuronal death of stroke]. PMID- 17461134 TI - [Pathophysiology of atherothrombotic infarction]. PMID- 17461135 TI - [Artery-to-artery embolic cerebral infarction]. PMID- 17461137 TI - [The mechanism and pathology of cerebral aneurysm formation]. PMID- 17461136 TI - [Mechanisms and pathology of hypertensive intracerebral hemorrhage]. PMID- 17461138 TI - [Experimental cerebral infarct model--history, current status and future tasks]. PMID- 17461139 TI - [Pathophysiology of brain edema formation following stroke: recent advancement]. PMID- 17461140 TI - [Ischemic brain and endoplasmic reticulum stress]. PMID- 17461141 TI - [Angiotensin II-induced facilitation of hypoxic neuronal damage through PKCdelta activation]. PMID- 17461142 TI - [Reactive oxygen species and free radicals are involved in pathophysiology of brain ischemia]. PMID- 17461143 TI - [Nitric oxide in cerebral ischemia]. PMID- 17461144 TI - [Ischemic brain damage and glial activation]. PMID- 17461145 TI - [Kinetics of inflammatory cytokines during hyperacute phase of ischemic stroke]. PMID- 17461146 TI - [Homocysteine]. PMID- 17461147 TI - [Adiponectin]. PMID- 17461148 TI - [Lipoprotein (a) * remnant lipoproteins]. PMID- 17461149 TI - [Hyperinsulinemia, insulin resistance]. PMID- 17461150 TI - [Antiphospholipid syndrome]. PMID- 17461151 TI - [Hemostatic abnormalities]. PMID- 17461152 TI - [High-sensitivity C-reactive protein]. PMID- 17461153 TI - [Chlamydia pneumoniae infection and stroke]. PMID- 17461154 TI - [Patent foramen ovale]. PMID- 17461155 TI - [Gene polymorphism in cerebrovascular disease]. PMID- 17461156 TI - [Clinical diagnosis of acute stroke]. PMID- 17461157 TI - [3-3-9 Coma Scale and Revised Coma Scale]. PMID- 17461159 TI - [Japanese guideline for the management of stroke 2004--general management of stroke]. PMID- 17461158 TI - [JSS-H (Japan Stroke Scale: Higher Cortical Function)]. PMID- 17461160 TI - [Intracerebral hemorrhage]. PMID- 17461161 TI - [Guidelines for aneurysmal subarachnoid hemorrhage]. PMID- 17461162 TI - [Guideline for intravenous rt-PA therapy of patients with ischemic stroke]. PMID- 17461163 TI - [Toward the new Japanese Guidelines for the Management of Stroke 2007]. PMID- 17461164 TI - [Recent progress of diagnostic imaging in stroke]. PMID- 17461165 TI - [Multi slice CT (MSCT)]. PMID- 17461166 TI - [CT perfusion (CTP)]. PMID- 17461167 TI - [3D CT angiography (CTA)]. PMID- 17461168 TI - [CBF measurement with xenon-enhanced CT]. PMID- 17461169 TI - [CT criteria for thrombolysis in acute ischemic stroke]. PMID- 17461170 TI - [Diffusion weighted--and perfusion weighted-magnetic resonance imaging]. PMID- 17461171 TI - [MR angiography (MRA)]. PMID- 17461172 TI - [Diffusion tensor tractography of stroke: three dimensional relationship, quantitative analysis using FA and probability mapping of the pyramidal tract]. PMID- 17461173 TI - [Multimodal MR]. PMID- 17461174 TI - [MRI plaque imaging]. PMID- 17461175 TI - [Image fusion of three-dimensional digital subtraction angiography and magnetic resonance imaging in the assessment of cerebrovascular diseases]. PMID- 17461176 TI - [Measurements of brain function by using functional MRI and near-infrared light spectroscopy]. PMID- 17461177 TI - [MRS (MR spectroscopy)]. PMID- 17461178 TI - [3D angiography with FPD system]. PMID- 17461179 TI - [Development of the PET-CT system]. PMID- 17461180 TI - [3D-SSP analysis with SPECT data for vascular disease]. PMID- 17461181 TI - [Ultrasonography of neck arteries]. PMID- 17461182 TI - [Transoral carotid ultrasonography]. PMID- 17461183 TI - [Guideline of magnetoencephalographic studies for ischemic cerebrovascular diseases]. PMID- 17461184 TI - [Symptoms and signs in cerebral herniation]. PMID- 17461185 TI - [Stroke and headache]. PMID- 17461186 TI - [Stroke and vertigo]. PMID- 17461187 TI - [Poststroke epilepsy]. PMID- 17461188 TI - [Cognitive impairment after stroke]. PMID- 17461189 TI - [Central post-stroke pain]. PMID- 17461190 TI - [Stroke presenting dementia as its cardinal symptom]. PMID- 17461191 TI - [Current and future strategies of medical therapy for acute stroke]. PMID- 17461192 TI - [Internal therapy of the chronic stage of stroke]. PMID- 17461193 TI - [Current status and new horizon in surgical treatment of cerebrovascular disease]. PMID- 17461194 TI - [Strokology in interventional neuroradiology era--development of endovascular surgery supported by new devices]. PMID- 17461195 TI - [JPPP]. PMID- 17461196 TI - [PROGRESS trial]. PMID- 17461197 TI - [J-STARS (Japan statin treatment against recurrent stroke)]. PMID- 17461198 TI - [MELT Japan (MCA-Embolism Local Fibrinolytic Intervention Trial Japan)]. PMID- 17461199 TI - [JET study (Japanese EC-IC Bypass Trial)]. PMID- 17461200 TI - [ISTICH (International Surgical Trial in Intracerebral Hemorrhage)]. PMID- 17461201 TI - [CREST (Carotid Revascularization Endarterectomy versus Stent Trial)]. PMID- 17461202 TI - [Unruptured cerebral aneurysm study in Japan: method and registration status]. PMID- 17461203 TI - [Anti-brain-edema agents]. PMID- 17461204 TI - [Edaravone]. PMID- 17461205 TI - [Sodium ozagrel]. PMID- 17461206 TI - [Aspirin]. PMID- 17461207 TI - [Ticlopidine]. PMID- 17461208 TI - [Basic principal of the dual antithrombotic therapy achieved by cilostazol]. PMID- 17461209 TI - [Argatroban]. PMID- 17461210 TI - [T-PA]. PMID- 17461211 TI - [Urokinase]. PMID- 17461212 TI - [Nicergoline, ibudilast, ifenprodil tartrate]. PMID- 17461213 TI - [Orengedokuto]. PMID- 17461214 TI - [Choto-san]. PMID- 17461215 TI - [Fasudil hydrochloride]. PMID- 17461216 TI - [Ximelagatran]. PMID- 17461217 TI - [Future aspect of regenerative medicine for cerebral infarction]. PMID- 17461218 TI - [Gene therapy with GDNF]. PMID- 17461219 TI - [A cell therapy for stroke]. PMID- 17461221 TI - [Stroke and hypertension]. PMID- 17461220 TI - [Hematopoietic cytokine therapy for cerebral infarction]. PMID- 17461222 TI - [Stroke and diabetes mellitus]. PMID- 17461223 TI - [Hyperlipidemia]. PMID- 17461224 TI - [Atrial fibrillation]. PMID- 17461225 TI - [Cigarette smoking and risk of stroke]. PMID- 17461226 TI - [Alcohol drinking]. PMID- 17461227 TI - [Obesity]. PMID- 17461228 TI - [Metabolic syndrome]. PMID- 17461229 TI - [Calcium channel blocker]. PMID- 17461231 TI - [Thiazolidinediones]. PMID- 17461230 TI - [Angiotensin converting enzyme inhibitors and angiotensin II type 1 receptor blockers]. PMID- 17461232 TI - [Role of HMG-CoA inhibitor (statin) in prevention of cerebral infarction]. PMID- 17461233 TI - [New development in stroke rehabilitation]. PMID- 17461234 TI - [Indication, evaluation and prognosis of acute stroke rehabilitation]. PMID- 17461235 TI - [Stroke rehabilitation--functional evaluation and rehabilitation approach]. PMID- 17461236 TI - [Clinical application of gait assistive robot training for hemiplegic patients]. PMID- 17461237 TI - [Rehabilitation for pain in poststroke patient]. PMID- 17461238 TI - [Sub-acute rehabilitation hospital--merits of in-patient care]. PMID- 17461239 TI - [Strategies for stroke rehabilitation based on neuroscience]. PMID- 17461240 TI - [Music therapy for cerebrovascular disease]. PMID- 17461241 TI - [Stroke center, its importance and expectation from it]. PMID- 17461242 TI - [Stroke unit and stroke care unit in a system of care in the community]. PMID- 17461243 TI - [Rehabilitation hospital: trends, missions, and issues]. PMID- 17461244 TI - [The present situation and a role of the convalescent rehabilitation ward]. PMID- 17461245 TI - [In-home rehabilitation]. PMID- 17461246 TI - [Referral system for stroke]. PMID- 17461247 TI - [Clinical pathway for stroke]. PMID- 17461248 TI - [Role of specialist in stroke managements in Japan]. PMID- 17461249 TI - [Public education campaign by the Japan Stroke Association and stroke awareness week]. PMID- 17461250 TI - [Brain Attack Campaign in United States]. PMID- 17461251 TI - Educational practices and assessment. PMID- 17461252 TI - Preparing future teachers and doctoral-level leaders in deaf education: meeting the challenge. AB - The deaf education profession faces a critical juncture. First, the 2006 leadership crisis that swept deaf education's flagship institution--Gallaudet University--will propel professionals to think deeply about promoting diversity, equity, and access in deaf education teacher and leadership preparation programs. Second, personnel shortages require attention: Teacher and leadership voids in university and K-12 programs loom if training efforts are not increased. Teaching and leadership needs center on three challenges: (a) understanding the changing demographic composition of the student, teacher, and leadership populations; (b) developing an evolving curriculum founded on research-based practices; (c) continuing to enlarge the knowledge base through applied research in the social sciences. Two case studies examine teacher training and leadership programs at universities that address these challenges. The importance of workplace deaf hearing bicultural teams is examined. Implications for the preparation of teacher and leadership personnel in deaf education are discussed. PMID- 17461253 TI - A comparative perspective on the experiences of deaf and hard of hearing individuals as students at mainstream and special schools. AB - Integration of individuals categorized as having special educational needs in mainstream schools has become a dominant policy in many countries. Changes in recent years in the field traditionally called "special education" have significantly influenced the education of deaf and hard of hearing individuals. The movements against segregation and toward integration and, more recently, inclusion, have created the conditions for educational changes, not only in mainstreaming but in special education. The article brings to light the views and experiences of deaf and hard of hearing people as students at special schools and mainstream schools, in order to compare the two systems from the viewpoints of those involved and to explore the possible implications of these views and experiences for the development of the educational system in Cyprus regarding inclusive education. Particular attention is given to improvement of the education of deaf and hard of hearing children. PMID- 17461254 TI - The Los Angeles County response to child abuse and deafness: a social movement theory analysis. AB - There is increasingly strong evidence that children with disabilities are at higher risk for maltreatment when compared to children without disabilities. There is also concern about the adequacy of child welfare services for children and parents with disabilities, particularly those disabilities that result in a communication impairment. This article describes a successful community practice effort in Los Angeles County that resulted in the establishment of a comprehensive array of linguistically and culturally competent child abuse prevention and treatment services for the maltreated deaf child and for the deaf parent at risk for child abuse perpetration. Social movement theory is used to analyze a change effort that was developed and implemented by a broad coalition of members of the Deaf and hearing communities. Elements of the problem, social movement theory, the coalition, the change strategy, and the results are described. PMID- 17461255 TI - Deafness and attention in deaf children. AB - The study shows the differences between hearing parents and deaf instructors interacting with deaf children and directing their attention. Data were collected at home and at a service for special needs in Bristol, England. The mother or instructor was asked to play naturally with the child with the toys provided. When the child's attention was focused during their play, the mother or instructor had to try to direct the child's attention to each of the toys. The results suggest that both groups (mothers and instructors) were effective in directing attention to objects not in the immediate area of play; however, hearing mothers were more successful than deaf instructors. PMID- 17461256 TI - Demographic characteristics and rates of progress of deaf and hard of hearing persons receiving substance abuse treatment. AB - A lack of demographic information and data related to the achievement of short term goals during substance abuse treatment among persons who are deaf or hard of hearing dictated the need for the study. New York State maintains a database on all individuals who participate in treatment. Within this database, 1.8% of persons in treatment for substance use disorder (SUD) were also deaf or hard of hearing. As hypothesized, members of the deaf and hard of hearing sample were older, likelier to be white, and likelier to be female, relative to the SUD-only group. For both groups, alcohol, heroin, and cocaine had the highest rates of reported use. Achievement of short-term goals in the areas of alcohol use, drug use, vocational/educational goals, and overall goals indicated no differences between the deaf and hard of hearing group and the SUD-only group. Implications of these findings are discussed. PMID- 17461257 TI - Googling "deaf": deafness in the world's English-language press. AB - An internet search tool, Google Alert, was used to survey the global English language press July-December 2005 for references to deaf people. The survey found that such references focus on people who are deaf rather than the disability itself, thus demonstrating how well deaf people fit into the mainstream. Derogatory terminology such as "deaf and dumb" was rare. However, when used metaphorically, the term deaf usually had negative connotations. Implications for the public view of deaf people are considered in this context. PMID- 17461258 TI - Where is it? How deaf adolescents complete fact-based internet search tasks. AB - An exploratory study was designed to describe Internet search behaviors of deaf adolescents who used Internet search engines to complete fact-based search tasks. The study examined search behaviors of deaf high school students such as query formation, query modification, Web site identification, and Web site selection. Consisting of two fact-based search tasks, the study was done in four regional day school programs for the deaf. As students conducted two search tasks, they completed task analyses of the selected Web sites and gave reasons for their selections. The research also identified the processes used by deaf students to compensate for limited English reading abilities while navigating search engines results that were typically written well above deaf students' average reading level. The results demonstrated that deaf adolescents were unable to initiate, conduct, analyze, or validate effective Internet searches in response to fact based search tasks. PMID- 17461259 TI - The proper conduct of research. AB - Scientific misconduct has garnered recent attention by the media over scandals concerning falsification and fabrication of data surrounding potentially promising breakthroughs in stem-cell research, allegations of plagiarism at a U.S. university, and financial conflicts of interest between researchers and drug companies. While this makes for interesting copy, discussion of scientific fraud provides an excellent opportunity to review ethical standards for research and examine the conflicts that confront researchers today. This review specifically focuses on five areas that involve scientific integrity-plagiarism, falsification, fabrication, authorship, and conflict of interest-as well as nuances in each area that even senior investigators may not be aware of (e.g., self-plagiarism). The standards for ethical conductance of research discussed in this review are those set by many scientific, peer-reviewed journals and by federal and private granting agencies, and therefore it highlights the expectations and guidelines surrounding manuscript and grant submissions and review, and the consequences associated with violations. This review is intended to stimulate discussion among readers and assess what is necessary to become a good, competitive, but ethical researcher, especially in an era of shrinking financial resources for research. PMID- 17461260 TI - Chlamydiae and atherosclerosis: can psittacine cases support the link? AB - Atherosclerosis is a common disease in pet birds, particularly in psittacines. Little is known about the role of risk factors predisposing birds to this disease. In our study, we tried to detect chlamydiae in formalin-fixed and paraffin-embedded atherosclerotic tissue from 103 pet birds to clarify their role in atherosclerosis. Methods used were polymerase chain reaction (PCR), sequencing, and immunohistochemistry. Histopathologic examination served to classify the extent of atherosclerotic lesions. In the PCR, 4 (3.9%) of 103 cases, all of them with advanced stages of atherosclerosis, were positive. Subsequent sequence analysis revealed high identities (94%-100%) with Chlamydophila psittaci in three cases. Interestingly, two of these birds came from C. psittaci-infected populations. Because of the low incidence (3.9%), the occurrence only in advanced stages, and the association with C psittaci-infected avian populations, a causal relationship between chlamydiae and atherosclerosis in pet birds is rather improbable. PMID- 17461261 TI - Bilateral effects of vaccination against infectious bursal disease and Newcastle disease in specific-pathogen-free layers and commercial broiler chickens. AB - Different infectious bursal disease virus (IBDV) live vaccines (intermediate, intermediate plus) were compared for their immunosuppressive abilities in specific-pathogen-free (SPF) layer-type chickens or commercial broilers. The Newcastle disease virus (NDV) vaccination model was applied to determine not only IBDV-induced immunosuppression but also bilateral effects between IBDV and NDV. None of the IBDV vaccines abrogated NDV vaccine-induced protection. All NDV vaccinated SPF layers and broilers were protected against NDV challenge independent of circulating NDV antibody levels. Sustained suppression of NDV antibody development was observed in SPF layers, which had received the intermediate plus IBDV vaccine. We observed a temporary suppression of NDV antibody development in broilers vaccinated with one of the intermediate, as well as the intermediate plus, IBDV vaccines. Different genetic backgrounds, ages, and residual maternal antibodies might have influenced the pathogenesis of IBDV in the different types of chickens. Temporary suppression of NDV antibody response in broilers was only seen if the NDV vaccine was administered before and not, as it was speculated previously, at the time the peak of IBDV-induced bursa lesions was detected. For the first time, we have demonstrated that the NDV vaccine had an interfering effect with the pathogenesis of the intermediate as well as the intermediate plus IBDV vaccine. NDV vaccination enhanced the incidence of IBDV bursa lesions and IBDV antibody development. This observation indicates that this bilateral effect of an IBDV and NDV vaccination should be considered in the field and could have consequences for the performance of broiler flocks. PMID- 17461262 TI - Anticoccidial effects of xanthohumol. AB - Xanthohumol (XN), a prenylated chalcone from the hops flower, was examined for its ability to reduce invasion of Madin-Darby bovine kidney (MDBK) cells by Eimeria tenella sporozoites (SZ), as well as to reduce invasion by E. tenella and E. acervulina SZ in the chick host. Additionally, XN was tested as an anticoccidial feed additive at 20 ppm against challenge infections with E. acervulina, E. maxima, and E. tenella. Cell invasion by E. tenella SZ was inhibited 66% by treatment of SZ with 22 ppm XN. This inhibition was associated with an apparent physical disruption of the apical ends of the SZ. Rectal challenges with E. tenella SZ treated with 5, 10, and 20 ppm XN resulted in significantly reduced gross-lesion scores and normal chick-host weight gains compared with challenge with untreated SZ. Oral challenges with similarly treated E. acervulina SZ, accomplished with prior antacid treatment, resulted in significantly reduced gross lesions and reduced oocyst shedding compared with challenge with untreated SZ and were associated with physical disruption of sporozoite morphology. In a pilot test, provision of feed supplemented with 20 ppm XN for 3 days before challenge to 6 days after challenge did not control challenge infections with E. acervulina, E. maxima, or E. tenella as judged by measurements of weight gain, feed conversion, and gross lesions. Although XN-fed chicks infected with E. acervulina and E. maxima shed fewer oocysts than those on control feed, the differences in numbers were not statistically significant (P > 0.05). PMID- 17461263 TI - Genetic characterization, pathogenicity, and protection studies with an avian adenovirus isolate associated with inclusion body hepatitis. AB - An avian adenovirus (AAV) was isolated from liver samples of two 2-wk-old broiler breeder flocks obtained from grandparents vaccinated at 10 and 17 wks of age with an autogenous inactivated vaccine containing the European AAV 8 (8565 strain) and 11 (1047 strain) serotypes (AAV8/11 vaccine). Affected broiler-breeders exhibited clinical signs and macroscopic and microscopic lesions associated with inclusion body hepatitis (IBH). The isolated adenovirus, identified as Stanford, was molecularly characterized as European serotype 9. The pathogenicity of the Stanford strain was confirmed after inoculation of specific-pathogen-free (SPF) chickens at 1-7 days of age, causing 100% and 20% mortality, respectively. The level of protection against IBH was evaluated in two broiler-breeder progenies from AAV 8/11-vaccinated grandparent flocks and a commercial broiler flock by challenge at 1 or 7 days of age with the AAV 8 and 11 serotypes and/or the Stanford strain. The broiler-breeder progenies and the commercial broiler flock exhibited protection against IBH after challenge. No significant differences in mean body weights were observed at 3 wk of age in any of the evaluated groups. We conclude that broiler-breeder progenies from 30- to 50-wk-old grandparents vaccinated with the AAV 8/11 vaccine were adequately protected against challenge with the AAV 8 and 11 serotypes and the Stanford strain. PMID- 17461264 TI - Pathogenicity and transmission of reticuloendotheliosis virus isolated from endangered prairie chickens. AB - The pathogenicity and transmission of a field isolate of reticuloendotheliosis virus (REV) was studied using an experimental model in Japanese quail. Oncogenicity was also evaluated after inoculations in chickens and turkeys. The original REV (designated APC-566) was isolated from Attwater's prairie chickens (Tympanuchus cupido attwateri), an endangered wild avian species of the southern United States. The transmissibility of the REV isolate was studied in young naive Japanese quail in contact with experimentally infected quail. Vertical transmission was not detected by virus isolation and indirect immunofluorescence. Seroconversion was detected in few contact quails, suggesting horizontal transmission. The APC-566 isolate induced tumors beginning at 6 wk of age in quails infected as embryos. Most of the tumors detected in Japanese quail were lymphosarcomas, and 81% of these neoplasias contained CD3+ cells by immunoperoxidase. REV APC-566 was also oncogenic in chickens and turkeys infected at 1 day of age, with tumors appearing as early as 58 days after infection in chickens and at 13 wk of age in turkeys. This study was conducted in part as an attempt to understand the potential for pathogenicity and transmission of REV isolated from endangered avian species. PMID- 17461265 TI - Colonization of specific regions of the reproductive tract and deposition at different locations inside eggs laid by hens infected with Salmonella enteritidis or Salmonella heidelberg. AB - Internal contamination of eggs by Salmonella Enteritidis has been a significant source of human illness for several decades and is the focus of a recently proposed U.S. Food and Drug Administration regulatory plan. Salmonella Heidelberg has also been identified as an egg-transmitted human pathogen. The deposition of Salmonella strains inside eggs is apparently a consequence of reproductive tissue colonization in infected laying hens, but the relationship between colonization of specific regions of the reproductive tract and deposition in different locations within eggs is not well documented. In the present study, groups of laying hens were experimentally infected with large oral doses of Salmonella Heidelberg, Salmonella Enteritidis phage type 13a, or Salmonella Enteritidis phage type 14b. For all of these isolates, the overall frequency of ovarian colonization (34.0%) was significantly higher than the frequency of recovery from either the upper (22.9%) or lower (18.1%) regions of the oviduct. No significant differences were observed between the frequencies of Salmonella isolation from egg yolk and albumen (4.0% and 3.3%, respectively). Some significant differences between Salmonella isolates were observed in the frequency of recovery from eggs, but not in the frequency or patterns of recovery from reproductive organs. Accordingly, although the ability of these Salmonella isolates to colonize different regions of the reproductive tract in laying hens was reflected in deposition in both yolk and albumen, there was no indication that any specific affinity of individual isolates for particular regions of this tract produced distinctive patterns of deposition in eggs. PMID- 17461266 TI - Heparan sulfate is a selective attachment factor for the avian coronavirus infectious bronchitis virus Beaudette. AB - The avian coronavirus infectious bronchitis virus (IBV) strain Beaudette is an embryo-adapted virus that has extended species tropism in cell culture. In order to understand the acquired tropism of the Beaudette strain, we compared the S protein sequences of several IBV strains. The Beaudette strain was found to contain a putative heparan sulfate (HS)-binding site, indicating that the Beaudette virus may use HS as a selective receptor. To ascertain the requirements of cell-surface HS for Beaudette infectivity, we assayed for infectivity in the presence of soluble heparin as a competitor and determined infectivity in mutant cell lines with no HS or glycosaminoglycan expression. Our results indicate that HS plays a role as an attachment factor for IBV, working in concert with other factors like sialic acid to mediate virus binding to cells, and may explain in part the extended tropism of IBV Beaudette. PMID- 17461267 TI - Severe nonpurulent encephalitis with mortality and feather lesions in call ducks (Anas platyrhyncha var. domestica) inoculated intravenously with H5N1 highly pathogenic avian influenza virus. AB - One-day-old, 2-wk-old, and 4-wk-old call ducks (Anas platyrhyncha var. domestica) inoculated intravenously with the H5N1 highly pathogenic avian influenza virus A/chicken/Yamaguchi/7/2004 isolate (Ck/Yama/7/04) were examined clinically, pathologically, and virologically. Clinically, the birds exhibited mild-to-severe neurologic signs and corneal opacity. All birds in the 1-day-old group and one bird in the 4-wk-old group died within 4 days after the virus inoculation. Histologic changes were characterized by severe nonpurulent encephalitis and necrotic lesions of feather epithelium on day 3 postinoculation (PI) or later. Focal necrosis of myocardial cells, pancreatic acinar cells, skeletal myocytes, and corneal epithelial cells was observed. Viral antigens were detected in association with necrotic changes. Viruses were isolated from all examined organs including the skin with many feathers. Serum antibody against the virus was detected in all surviving birds on day 10 PI by hemagglutination-inhibition tests. These results suggest that Ck/Yama/7/04 has a pathogenicity that causes neurologic sign, nonpurulent encephalitis with mortality, and feather lesions for call ducks. Feather lesions with viral antigens and the virus isolation from the skin suggest that Ck/Yama/ 7/04 has a predilection for feathers in call ducks. PMID- 17461268 TI - Experimental reproduction of transmissible viral proventriculitis by infection of chickens with a novel adenovirus-like virus (isolate R11/3). AB - Transmissible viral proventriculitis (TVP) was experimentally reproduced in 2-wk old specific-pathogen-free chickens and commercial broiler chickens by eyedrop inoculation of adenovirus-like virus (AdLV), isolate R1 1/3. No clinical signs and no weight gain depression were observed in chickens inoculated with AdLV (R11/3); however, gross and microscopic lesions characteristic of TVP were present in proventriculi of inoculated chickens. Proventriculi of AdLV (R11/3) inoculated chickens were markedly enlarged, compared with sham-inoculated controls, by day 7 postinoculation (PI). Microscopic lesions in proventriculi of inoculated chickens were detected beginning on day 3 PI and consisted of degeneration and necrosis of glandular epithelium, ductal epithelial hyperplasia, replacement of glandular epithelium with ductal epithelium, and diffuse interstitial lymphoid infiltration; no microscopic lesions were observed in other tissues. AdLV (R11/3) antigens were detected in proventriculi by immunohistochemistry on days 3-10 PI in inoculated SPF chickens and days 3-21 PI in inoculated commercial broiler chickens; no viral antigens were detected in other tissues. AdLV (R11/3) was reisolated from proventriculi of inoculated SPF and commercial broiler chickens on days 5 and 7 PI. No virus, viral antigens, or lesions were detected in proventriculi collected from sham-inoculated chickens. These findings indicate an etiologic role for AdLV (R11/3) in TVP. PMID- 17461269 TI - Biological characteristics of chicken anemia virus regenerated from clinical specimen by PCR. AB - Our previous genetic characterization of chicken anemia virus (CAV) in commercial broiler chickens in Alabama revealed a previously undetected polymorphism: a glutamine codon at VP1 position 22, in 7 of the 14 sequences. The novel glutamine codon was always found in association with a VP1 "hypervariable region" identical to CAV field isolates that replicate poorly in culture. The complete genome of CAV73, representative of the sequences with the novel polymorphism, was generated from cloned polymerase chain reaction (PCR) fragments amplified directly from naturally infected tissues. CAV73 had been detected in 31-day-old broilers submitted for examination for reasons unrelated to anemia. After electroporation of the cloned genomes into MDCC-CU147 lymphoblastoid cells, the regenerated CAV caused the culture to fail within 9 days, and the medium contained 5 X 10(6) TCID50 CAV/ml. Use of MDCC-CU147 cells was essential, as identical electroporation of MDCC-MSB1 cells failed to generate CAV able to destroy the culture within 8 wk. Regenerated CAV73 produced anemia and severe lymphocytic depletion of the thymus when inoculated into susceptible 3-day-old chickens and was reisolated from these chickens. Furthermore, it replicated in low- and high passage MDCC-MSB1 cells similarly to a low-passage CAV field isolate that contains a different VP 1 "hypervariable region." The regeneration of CAV from PCR products directly from naturally infected carcasses, as performed in this study, provides a tool for the evaluation of distinct genetic polymorphisms that may be detected in specimens where infective virions are no longer available. Our results also provide some insight into the differential susceptibility of cell lines for low-passage CAV field isolates. PMID- 17461270 TI - Protection of chickens against a lethal challenge of Escherichia coli by a vaccine containing CpG oligodeoxynucleotides as an adjuvant. AB - Synthetic oligodeoxynucleotides (ODN) containing cytosine-phosphodiester-guanine (CpG) motifs (CpG-ODN) have been shown to be effective immunoprotective agents and vaccine adjuvants in a variety of bacterial, viral, and protozoan diseases in different animal species. The objective of this study was to compare the immune response of chickens to a killed Escherichia coli vaccine combined with oil in water emulsion or with CpG-ODN. Birds were vaccinated with killed E. coli antigens with either 10 or 50 microg of CpG-ODN on days 10 and 20 of age. At day 30, a virulent isolate of homologous E. coli was applied on a scratch site on the caudal abdominal region. Birds were examined for 10 days post-E. coli challenge, and pathologic and bacteriologic assessments were conducted on all birds that were either found dead or euthanized. The E. coli vaccine group that received no CpG-ODN had a survival rate of 65%. In contrast, groups that received the vaccine with CpG-ODN adjuvant had significantly higher survival rate of 92% (P < 0.01) with isolation of low numbers of E. coli from internal organs. Total IgG against E. coli antigens was highest in groups that received CpG-ODN as an adjuvant. Birds that received vaccine containing CpG-ODN had minimal inflammatory reaction without tissue necrosis at the injection site. Severe tissue necrosis was present in birds that received vaccine containing oil in water emulsion adjuvant. This study demonstrated that CpG-ODN is an effective vaccine adjuvant in chickens and results in minimal tissue destruction. This study is the first study in which CpG ODN has been demonstrated to produce an adaptive immune response, at a significant level, against an extracellular bacterial infection in chickens. PMID- 17461271 TI - Identification and expression of a gene encoding an epitope that induces hemagglutination inhibition antibody to Avibacterium paragallinarum serovar A. AB - The aims of this study were the identification, cloning, and expression of a genetic region encoding an epitope that induces hemagglutination inhibition (HI) antibody against Avibacterium paragallinarum serovar A and an evaluation of the recombinant protein for immunogenicity in chickens. Although two monoclonal antibodies (MAbs) with HI activity, designated S24-951 and S7-1716-5C, were generated in this study, no reactive proteins with both MAbs were identified by Western blot analysis. A gene fragment of 5157 bp, designated hpa5. 1, was cloned from genomic DNA, and a recombinant protein expressed by hpa5.1, designated HPA5.1, reacted with both MAbs on dot-blot analysis. HPA5.1 showed no hemagglutinating activity, but significantly absorbed HI antibodies in the chicken immune serum. Analysis using a series of deletion mutants prepared from hpa5.1 indicated that a 4.8 kbp gene in hpa5.1 is essential for the expression epitope recognized by MAb S24-951. In addition, chickens immunized once with HPA5.1 showed a high protection rate with sufficient HI antibody titers against challenge exposure with a virulent strain of A. paragallinarum serovar A strain 221. These results show that hpa5. I1 is responsible for the expression of an epitope that induces HI antibody, and HPA5.1 might be a candidate for the development of a new vaccine against avian infectious coryza caused by A. paragallinarum serovar A. PMID- 17461272 TI - Genetic analysis of a duck circovirus detected in commercial Pekin ducks in New York. AB - The genetic organization of the duck circovirus (DuCV) 33753-52 detected in commercial Pekin duck flocks from Long Island, NY, is described. The nucleotide sequence of virus 33753-52 exhibited high similarity with DuCVs previously detected in Germany and Hungary. It is possible that this DuCV from New York shares the same ancestor with the European counterparts. The virus 33753-52 exhibited genetic features characteristic of other circoviruses, such as the presence of two major open reading frames (rep and cap), two intergenic regions, one stem-loop structure, four intergenic direct repeats, and the conserved motifs for the rolling circle replication and for the dNTP binding domain in the Rep protein. This report is the first report of the presence of DuCV in commercial Pekin duck farms in the United States. The clinical and pathologic significance of DuCV in the duck farms located on Long Island needs to be clarified. DuCv was detected in culled birds, due to low body development, leg deformities, or arthritis. Staphylococcus aureus and Riemerella anatipestifer serotype 4 were isolated from some of the DuCV-positive birds. The apparent low prevalence of the virus suggests that at this time, this infection is not a significant problem for the duck industry in New York. However, the immunosuppressive properties of this virus need to be clarified as well as its role as a predisposing agent for other diseases. PMID- 17461273 TI - Performance evaluation of five detection tests for avian influenza antigen with various avian samples. AB - In this paper, we report on the evaluation of five influenza antigen detection tests by avian influenza H5N 1 virus-positive swab samples to estimate their diagnostic sensitivity. The tests included two chromatographic immunoassays, an H5 avian influenza-specific antigen detection enzyme-linked immunosorbent assay (ELISA), an influenza A antigen detection ELISA, and an H5 rapid immunoblot assay. The results showed that the overall sensitivities of these tests ranged from 36.3% to 51.4% (95% confidence interval ranging from 31.0% to 57.0%), which were comparable to Directigen Flu A antigen detection tests but substantially lower than genome detection methods. Diagnostic sensitivity performance is a function of the concentration of antigens in samples and the analytical sensitivity of the individual test. The test sensitivities were significantly higher for sick and dead birds by cloacal, tracheal, or tissue swabs than for fecal swabs from apparently healthy birds, and these tests would not be suitable for surveillance testing of clinically healthy birds. Furthermore, the sensitivity for testing tracheal and cloacal swabs from waterfowl and wild birds was not as good as for chickens. This was most likely to be associated with variation in virus titers between specimens from different bird species. However, the tests showed good sensitivities for testing brain swabs from clinically affected waterfowl species. The results indicate that these antigen detection tests could be used for preliminary investigations of H5N 1 outbreaks as a low cost, simple flock test in sick and dead birds for the rapid detection of H5N1 infection. However, the relatively low sensitivity of the tests as individual bird tests means that they should be used on optimal clinical specimens from diseased birds, testing birds on a flock basis, or testing samples as close to the onset of disease as possible before viral titers diminish. They should be followed up by confirmatory tests, such as reverse transcription polymerase chain reaction or viral culture, wherever possible but could assist in facilitating rapid investigations and control interventions. PMID- 17461274 TI - Experimental infection of chickens and turkeys with Mycoplasma gallisepticum reference strain S6 and North Carolina field isolate RAPD type B. AB - During an epidemic of mycoplasmosis in chicken and turkey flocks in North Carolina between 1999 and 2001, isolates of Mycoplasma gallisepticum (MG) from affected flocks were characterized by random amplification of polymorphic DNA (RAPD), and eight distinct RAPD types were identified. MG RAPD type B accounted for more than 90% of the isolates and was associated with moderate-to-severe clinical signs and mortality. The virulence of MG RAPD type B for chickens and turkeys was compared with sham-inoculated negative controls and MG S6 (a virulent strain)-inoculated positive controls. Clinical signs occurred in chickens and turkeys inoculated with either MG RAPD type B or MG S6. However, they were not as frequent or severe as those seen in naturally affected flocks, and there was no mortality in the experimental groups. Based on gross and microscopic findings, MG RAPD type B was equal to or more virulent than MG S6. All MG-inoculated birds were culture and PCR positive at 7 and 14 days postinoculation (PI). Among serological tests, the serum plate agglutination test was positive for the majority of chickens and turkeys (58%-100%) infected with either strain of MG at both 7 and 14 days PI. The hemagglutination inhibition test was negative for all birds at 7 days PI and positive for a few chickens (8%-17%) and several turkey sera (40%-60%) at 14 days PI. Only a single serum was positive by enzyme-linked immunosorbent assay (an MG S6-infected turkey) at 14 days PI. PMID- 17461275 TI - Comparative investigation of prevalence and antimicrobial resistance of Salmonella between pasture and conventionally reared poultry. AB - The objective of this study was to compare Salmonella prevalence and antimicrobial resistance between pasture and conventional poultry farms. We collected the first Salmonella prevalence and antimicrobial resistance data on pasture poultry farms. Fecal droppings were collected from 31 farms from Wisconsin (nine farms from each production type) and the Southeast (North Carolina, Virginia, and South Carolina; five conventional and 10 pasture poultry farms) in a 1-yr period. The specimens were cultured for Salmonella and tested for resistance to 12 antimicrobials. A univariate analysis was conducted to determine the significant differences in prevalence and resistance. At the farm level, no significant difference in Salmonella prevalence was found on 33% pasture and 47% conventional poultry farms (P= 0.4928). On an individual specimen level, flocks reared conventionally had higher prevalence than in pasture (P < 0.0001). Of all the isolates found to have resistance, 80% were from the Southeast. Of all the pasture isolates tested, 5% (8 of 162) were resistant to ceftriaxone, which is higher than previously reported from any production setting. None of the isolates from conventional flocks showed resistance to ceftriaxone. Multidrug resistance (resistance to three or more classes of antimicrobials) was found in 69% of the isolates from conventional farms and 11% on pasture farms in the Southeast (P < 0.0001), with the predominant resistance type of AmCSSuTeAx (ampicillin, chloramphenicol, streptomycin, sulfasoxazole, tetracycline, amoxicillin/clavulanic acid; 62%). About 5% of the pasture isolates from the Southeast showed the AmAxCFCRO (ampicillin, amoxicillin/clavulanic acid, cephalothin, ceftriaxone) multidrug resistance pattern. None of the isolates from Wisconsin were found to be multidrug resistant. PMID- 17461276 TI - FTA liver impressions as DNA template for detecting and genotyping fowl adenovirus. AB - The feasibility of using liver impressions on Flinders Technology Associates (FTA filter paper for the collection, inactivation, and molecular analysis of fowl adenovirus (FAV) was evaluated. FAV I European Union (EU) serotype 1 spotted on FTA was shown to be inactivated using specific-pathogen-free (SPF) primary chicken embryo liver cell culture as indicated by absence of cytopathic effect. Sensitivity of the polymerase chain reaction (PCR) test using tenfold dilutions of allantoic fluid from 100 to 10-4 for the detection of adenovirus serotype 1 on FTA cards was determined to be 0.0005 mean tissue culture infectious dose per FTA spot. The stability of the DNA from liver impressions on the FTA was found to be 198 days when stored at -20 degrees C. In a trial, inclusion body hepatitis (IBH) was experimentally reproduced in SPF chickens inoculated with FAV I EU serogroup 1, 4, 8, or 11, which presented weakness, pallor, depression, dehydration, and mortality within 6 days after inoculation. PCR performed on FTA liver impressions from the inoculated birds was able to detect all four viruses, and the nucleotide sequence analysis of the amplified PCR products (1219 bp of the hexone gene) revealed the expected serotypes. In addition to the trial, 55 clinical samples were analyzed from liver impressions on FTA cards, and FAV was detected in 11 of 55 (20%). Sequencing analysis showed that the viruses were EU serotypes 4, 5, 9, and 10. The results demonstrate that FTA filter paper inactivates the FAV I and maintains the DNA template for molecular analysis. PMID- 17461277 TI - Selection for early (precocious) development of Eimeria meleagridis in the turkey. AB - A precocious line of Eimeria meleagridis was developed by repeated propagation of the parasite in young turkeys and collection of the very first oocysts produced following infection. After 20 generations of selection, the prepatent period was reduced by 4-8 hr. Weight gain of poults given this line, from days 0-3 after infection, was significantly greater than that of poults given the parental strain, indicating that selection resulted in a loss of pathogenicity. Poults immunized with the precocious line produced no oocysts following challenge with the parental strain, indicating that immunogenicity was retained following selection. This is the first published report of the selection of a precocious line of Eimeria in the turkey. PMID- 17461278 TI - West Nile virus antibody prevalence in American crows (Corvus brachyrhynchos) and fish crows (Corvus ossifragus) in Georgia, USA. AB - Crows have been the centerpiece of avian West Nile virus (WNV) surveillance and research in North America. This work has demonstrated variation in susceptibility to WNV infection between American (Cor vus brachyrhynchos) andFish Crows (Corvus ossifragus). The higher WNV-associated mortality rate in American Crows compared with Fish Crows suggests that WNV antibody prevalence would be greater in the Fish Crow population. The objectives of this study were to 1) determine whether Fish Crows had higher WNV antibody prevalencethan American Crows, 2 ) determine th e persistence o f antibodies to WNV in naturally infected Fish Crows, and 3) develop a technique to distinguish Fish Crows from American Crows on the basis of sequence analysis and restriction enzyme digestion of a mitochondrial DNA fragment. West Nile virus antibody prevalence was 16.5% (n = 97) in Fish Crows and 5.7% in American Crows (n = 53) collected from Georgia between 2004 and 2006. Antibodies persisted at high titers for 12 mo in Fish Crows. This is the first report of WNV antibody persistence in a crow species. A polymerase chain reaction technique paired with restriction enzyme digestion easily distinguished American Crows from Fish Crows on the basis of a mitochondrial DNA fragment. PMID- 17461279 TI - Impact of different husbandry conditions on contact and airborne transmission of H5N1 highly pathogenic avian influenza virus to chickens. AB - Typically highly pathogenic avian influenza (HPAI) viruses spread very rapidly among chickens within sheds. However, the spread was slower than expected for the initial 10 days of the index farm in Japan during 2004. This slow spread, as well as the lack of gross lesions, clinical signs, or high mortality, hindered the field veterinarian from reporting a suspected HPAI outbreak to the veterinary office. To understand the field conditions for the slow virus spread, we examined contact and airborne transmission of the H5N1 virus to chickens in a negative pressure isolator using various numbers of infected chickens and separate compartments. We found that the contact transmission did occur inefficiently when one or two chickens were infected, whereas the transmission was efficient when four chickens were infected. Airborne transmission of the HPAI virus was also dependent on the number of infected chickens and was less efficient than contact transmission. These data together with field observations suggested that number of infected chickens, chicken house types, and amount of environmental contamination might affect the virus transmission efficiency to chickens. PMID- 17461280 TI - Retroviral inclusions in the enteric smooth muscle of a tumor-bearing young chicken. AB - A 15-cm segment of small intestine from a 7-wk-old broiler chicken presented for slaughter was encased by a firm, white mass. Other tissues were grossly unremarkable. Microscopically, the enteric serosa and peripheral muscularis of this segment of small intestine were replaced by a fibrosarcoma. Numerous linear, intracytoplasmic, eosinophilic inclusion bodies were present in smooth muscle cells of the muscularis of the small intestine, and a few similar inclusions were present in the muscularis of the proventriculus. In the heart, there were rare intracytoplasmic inclusions typical of viral matrix inclusions. Ultrastructurally, inclusion bodies in enteric smooth muscle were viral matrix inclusions, and virions resembling avian retroviruses were present in adjacent intercellular spaces. Polymerase chain reaction (PCR) of the deoxyribonucleic acid (DNA) extracted from tumor tissues indicated the presence of proviral DNA of subgroup J avian leukosis virus. This is the first description of the light microscopic appearance of these viral matrix inclusions in enteric smooth muscle. PMID- 17461281 TI - Serological survey on prevalence of antibodies to avian paramyxovirus serotype 2 in China. AB - We report the prevalent status of avian paramyxovirus serotype 2 (APMV-2) in China. Between 2003 and 2005, 9156 sera in total were collected and screened for APMV-2 antibodies by using the hemagglutination inhibition assay. The averaged seropositivity ofAPMV-2 for chickens, ducks, peacocks, ostriches, and partridges was 42.9%, 25.1%, 45.8%, 47.6%, and 80.0%, respectively. The results of this survey indicate that the distribution of APMV-2 is very widespread in China and that more attention should be paid to the influence of APMV-2 on poultry production. PMID- 17461282 TI - Clinical findings, lesions, and viral antigen distribution in great gray owls (Strix nebulosa) and barred owls (Strix varia) with spontaneous West Nile virus infection. AB - West Nile Virus (WNV) infection manifests itself clinically a nd pathologically differently in various species of birds. The clinicopathologic findings and WNV antigen tissue distribution of six great gray owls (Strix nebulosa) and two barred owls (Strix varia) with WNV infection are described in this report. Great gray owls usually live in northern Canada, whereas the phylogenetically related barred owls are native to the midwestern and eastern United States and southern Canada. Naturally acquired WNV infection caused death essentially without previous signs of disease in the six great gray owls during a mortality event. Lesions of WNV infection we re dominated by hepatic and splenic necrosis, with evidence o f disseminatedintravascular coagulation in the great gray owls. WNV antigen was widely distributed in th e organs of the great gray owls and appeared totarget endothelial cells, macrophages, and hepatocytes. The barred owls represented two sporadic cases. They had neurologic disease with mental dullness that led to euthanasia. These birds had mild to moderate lymphoplasmacytic encephalitis with glial nodules and lymphoplasmacytic pectenitis. WNV antigen was sparse in barred owls and only present in a few brain neurons and renaltubular epithelial cells. The cause of the different manifestations of WNV disease in these fairly closely related owl species is uncertain. PMID- 17461283 TI - Giant cell tumor of the bone in a scarlet macaw (Ara macao). AB - A 6-mo-old female scarlet macaw (Ara macao) was presented after a 2-mo period of anorexia and weakness. The bird was reluctant to fly 1 wk before referral due to a painful left wing. Physical examination revealed a firm swelling around the left shoulder. On radiographs, the diaphysis and proximal metaphysis of the left scapula were radiolucent. Computer tomography revealed an osteolytic process, suggestive of a bone tumor, affecting the left scapula. Cytology of a fine needle aspiration biopsy of the mass showed erythrocytes, a proliferation of spindle shaped mesenchymal cells, and multinucleated giant cells (osteoclasts) suggestive of a giant cell tumor. The left wing, including the scapula, was amputated. The bird showed a fast recovery but died 1 hr later. Findings during the pathological examination were compatible with shock due to blood loss. The shoulder process was characterized as a giant cell tumor. To our knowledge, this is the first complete report of a giant cell tumor of the bone in a bird. PMID- 17461284 TI - Chronic pulmonary interstitial fibrosis in a blue-fronted Amazon parrot (Amazona aestiva aestiva). AB - A 30-yr-old blue-fronted Amazon parrot (Amazon aestiva aestiva) was presented to the clinic with a history of sneezing more often during the last 2 mo. Physical examination revealed only a mild nasal discharge. Complete hematologic and plasma biochemical examination showed no abnormalities. Computerized tomography (CT) of the complete bird showed generalized lung alterations consistent with lung fibrosis. Two lung biopsies were taken. The results of the histologic examination of the biopsies confirmed the tentative CT diagnosis of pulmonary interstitial fibrosis. To our knowledge this is the first reported case of chronic pulmonary interstitial fibrosis diagnosed by means of a lung biopsy in an avian species. The histologic characteristics are discussed and compared with those of human idiopathic pulmonary fibrosis. PMID- 17461285 TI - Examining the usability of a virtual reality driving simulator. AB - The current study examined specific aspects of usability of a newly developed VR driver rehabilitation (VR-DR) system. Measures of user feedback and user comfort were examined among 54 participants: 33 individuals with acquired brain injury (20 with traumatic brain injury and 13 with cerebral vascular accident) and 21 healthy controls. All participants were administered the VR-DR and completed the VR-DR User Feedback Questionnaire. To examine group differences, a one-way analysis of variance was performed, comparing the user feedback total score between the three groups. The results indicated that the two clinical populations (traumatic brain injury and cerebral vascular accident) varied from the nonclinical population (healthy controls). A standard multiple regression analysis revealed that age was the only significant participant factor that contributed to the differences in user feedback ratings. Finally, consistent across the three groups, a distinct relationship was found between the self reported user rating and the onset of simulation sickness. The current findings indicate that individuals with traumatic brain injury and cerebral vascular accident provided less favorable user feedback ratings than healthy controls in the use of a new VR-DR system. This difference was not accounted for by differences in gender, education, or cognitive status and was only slightly accounted for by age. Delineating these various aspects of user feedback can assist in identifying potential confounds in VR-DR performance and help refine the application of the VR-DR for clinical decision making. PMID- 17461286 TI - Extending the evaluation of novel microswitch technology for small responses in children with profound multiple disabilities. AB - This study extended the evaluation of microswitch technology (i.e., a position sensor and an optic device), recently introduced for chin and eyelid responses, with two new participants with profound multiple disabilities. The participants were girls of 12.5 and 4.0 years of age who did not possess any specific response that they could profitably use in their environment. The study was conducted according to an ABAB design with a 3-week postintervention check. The results showed that both participants increased their level of responding during the intervention phases and postintervention check compared to the baseline phases. This positive outcome was discussed in terms of (a) the apparent suitability of the technology and responses used in the study, (b) the beneficial implications for the participants, and (c) new developments in the area. PMID- 17461287 TI - Intelligent systems in pediatric rehabilitation. PMID- 17461288 TI - Perspectives on mobile robots as tools for child development and pediatric rehabilitation. AB - Mobile robots (i.e., robots capable of translational movements) can be designed to become interesting tools for child development studies and pediatric rehabilitation. In this article, the authors present two of their projects that involve mobile robots interacting with children: One is a spherical robot deployed in a variety of contexts, and the other is mobile robots used as pedagogical tools for children with pervasive developmental disorders. Locomotion capability appears to be key in creating meaningful and sustained interactions with children: Intentional and purposeful motion is an implicit appealing factor in obtaining children's attention and engaging them in interaction and learning. Both of these projects started with robotic objectives but are revealed to be rich sources of interdisciplinary collaborations in the field of assistive technology. This article presents perspectives on how mobile robots can be designed to address the requirements of child-robot interactions and studies. The authors also argue that mobile robot technology can be a useful tool in rehabilitation engineering, reaching its full potential through strong collaborations between roboticists and pediatric specialists. PMID- 17461289 TI - Building Robota, a mini-humanoid robot for the rehabilitation of children with autism. AB - The Robota project constructs a series of multiple-degrees-of-freedom, doll shaped humanoid robots, whose physical features resemble those of a human baby. The Robota robots have been applied as assistive technologies in behavioral studies with low-functioning children with autism. These studies investigate the potential of using an imitator robot to assess children's imitation ability and to teach children simple coordinated behaviors. In this article, the authors review the recent technological developments that have made the Robota robots suitable for use with children with autism. They critically appraise the main outcomes of two sets of behavioral studies conducted with Robota and discuss how these results inform future development of the Robota robots and robots in general for the rehabilitation of children with complex developmental disabilities. PMID- 17461290 TI - [Image of the month. Systolic anterior motion after repair of the mitral valve]. PMID- 17461291 TI - [Image of the month. Double eggshell calcification]. PMID- 17461292 TI - [How I treat....A stoutness-associated intertrigo]. AB - Intertrigo is a common inflammatory disorder of various origins. It can represent a single skin manifestation or be part of a disorder possibly exhibiting specific manifestations on other parts of the body. Hence, intertrigo may represent a sign of a peculiar disease. It may also result from physicochemical aggressions of the skin following occlusion of the large skinfolds particularly in case of stoutness. In this intance, a treatment similar to that of diaper dermatitis can be offered. A paste enriched or not with miconazole nitrate is indicated to prevent or treat most presentations of intertrigo. PMID- 17461293 TI - [Clinical case of the month. Left ventricular apical ballooning (Tako-Tsubo cardiomyopathy)]. AB - A 80-year-old European woman was admitted for a cardiac syndrome which mimics acute coronary insufficiency, with apical akinesis and basal hyperkinesis. The first description of this syndrome (Tako-Tsubo cardiomyopathy) was reported in Japanese patients in the 90's. Recently this reversible left ventricular cardiomyopathy was also described in USA and Europe in white patients. This cardiomyopathy often develops in older women, after emotional or physical stress. The etiology remains unknown. PMID- 17461294 TI - [Clinical case of the month. Nutcracker syndrome in association with a painful nephrologic disease]. AB - Compression of the left renal vein between the aorta and the superior mesenteric artery causes a physiological condition, the so-called nutcracker phenomenon, but it can sometimes lead to left venous hypertension, or "nutcracker syndrome". Classical manifestations of which are an association of left flank pain, unilateral proteinuria and unilateral hematuria, without renal impairment. We report an atypical association of nutcracker syndrome with IgM nephropathy. PMID- 17461295 TI - [Clinical case of the month. Peritoneal tuberculosis mimicking ovarian carcinoma]. AB - Peritoneal tuberculosis can mimic advanced stage ovarian cancer and can lead to the performance of an unnecessary extended surgery. Clinical discrimination between peritoneal tuberculosis and ovarian carcinoma may sometimes be extremely difficult. We report 2 cases of peritoneal tuberculosis mimicking ovarian carcinoma. Diagnosis was made on perioneal biopsy. PMID- 17461296 TI - [Cardiometabolic effects of rimonabant in obese/overweight subjects with dyslipidaemia or type 2 diabetes]. AB - Rimonabant (Acomplia) is the first selective CB1 receptor blocker of the endocannabinoid system. It has been evaluated in the RIO ("Rimonabant In Obesity and related disorders") programme including above 6.600 overweight/obese patients with or without comorbidities followed for 1 to 2 years. Compared to placebo, rimonabant 20 mg/day consistently increases weight loss, reduces waist circumference, increases HDL cholesterol, lowers triglyceride levels, diminishes insulin resistance, and reduces the prevalence of metabolic syndrome. In patients with type 2 diabetes, rimonabant also diminishes HbA1c levels, an effect confirmed in the recent SERENADE trial. Almost half of the metabolic effects occurs beyond weight loss, suggesting direct peripheral effects of rimonabant. Rimonabant is indicated in Europe as an adjunct to diet and exercise for the treatment of obese patients, or overweight patients with associated risk factor(s), such as type 2 diabetes or dyslipidaemia. PMID- 17461297 TI - [Hypotension in the very preterm infant]. AB - One out of four very preterm infants will present with circulatory maladaptation during the first two days of life, with an increased risk of early complications and long term sequelae. Appreciation of those transitional difficulties cannot be limited to blood pressure. Assesment of blood pressure itself must be done in relation with gestational age and birth weight adapted norms. The effects of therapies for low systemic blood flow on blood pressure, organs and cerebral circulations are better understood, but none of them has assessed for mortality or neurodevelopmental outcomes. PMID- 17461298 TI - [Arsenical poisoning: how and why to diagnose it]. AB - Arsenic is an ubiquitary element which has been widely used for centuries in different fields such as medicine, agriculture or industry. Acute or chronic exposure to As can lead to various dermatological and systemic disorders with a possibe latency over decades. The dermatological signs of As intoxication are important to detect since one of the potential complications is carcinoma. PMID- 17461299 TI - [The question of secondary medical transfer in Belgium: the experience conducted at the University Hospital Center of Liege]. AB - Evaluation of the aid of an emergency mobile unit to transfer monitorized patients to a University hospital, in the political context of regional care network offering highly qualified but restricted tertiary area centres, and an open prospective study conducted over the 5 first months in 2006. The call regulation was assessed by the emergency physician of the transfer team and all missions were concluded with an evaluating report. An amount of 197 requests were taken into account from which 80 % were addressed between 8 am and 8 pm. The mean average time interval for missions was 59 minutes (base to base) and the distance covered was 20.7 km as a mean. In essence, indications for medical secondary transfer regarded patients in need for acute coronary care (42.6%), specific intensive care (26.4%) and neurosurgical interventions (19.3%). We noted that endotracheal intubation occurred in only one case. Mortality during such a transfer activity was absent. Medical transfer unit allows the development of specific high qualified network resources owing to the secondary addressee of patients. However, the weak incidence of complication questions the practice of systematic medical accompanying during such transfers. PMID- 17461300 TI - [When depression does not end. Resistant depression: recent clinical and therapeutic aspects]. AB - All over the world, depression represents a major public health issue. According to WHO, it is the fifth disease in the world (in years of illness), the second for people aged 15 to 44 and by 2020, it could become the second in the whole world population after cardiovascular diseases. Although increasingly safer and more efficient antidepressants are available, physicians are frequently confronted with the problem of resistant depression. Indeed, an estimated 30 to 45% of patients treated for major depressive disorder have either a partial response or no response at all to the antidepressive treatment. Residual symptoms are very common. Besides usual strategies such as lithium or thyroid hormones adding, some new approaches are now being used to improve the treatment of resistant depression. These are in particular pharmacological strategies such as switching or augmentation, psychotherapy and cerebral stimulation technology. This article reviews the latest clinical and therapeutic aspects of resistant depression. PMID- 17461301 TI - [Congestive heart failure and diabetes mellitus: an intricated relationship]. AB - The present article analyses the intimate relationship between diabetes mellitus and congestive heart failure. This relationship is indeed "bidirectional". On the one hand, it is well known that diabetes mellitus, especially type 2 diabetes, predisposes to congestive heart failure due to intricated mechanisms. In most cases, there is a combination of various well-known risk factors, such as obesity, arterial hypertension and coronary heart disease, with a specific diabetic cardiomyopathy, whose pathophysiology is complex. On the other hand, several studies showed that congestive heart failure increases the risk of new type 2 diabetes, probably due to decreased muscular perfusion and excessive neurohumoral response. Remarkably, such a risk could be reduced by a drug capable of inhibiting the renin-angiotensin system, as previously reported in patients with arterial hypertension. The recent launch and the increasing use of thiazolidinediones (glitazones) raise the interest for congestive heart failure in diabetic patients. Indeed, because of their positive effect on insulin resistance and various pleiotropic effects, glitazones may exert some cardiovascular protection; however, both rosiglitazone and pioglitazone are associated with fluid retention, which could reveal or aggravate episodes of congestive heart failure. PMID- 17461302 TI - [Anatomo-clinical comparison: caecal duplication cyst]. AB - Enteric duplication cysts, and particularly caecal duplication cysts, are rare and generally benign congenital anomalies for which a diagnosis is difficult to make because they mimic other surgical diseases. A surgical management is the treatment of choice. The diagnosis can then be made or confirmed by histopathologic analysis. PMID- 17461303 TI - [Drug of the month. Varenicline (Champix)]. AB - Smoking is responsible for 20,000 premature deaths per year in Belgium. Paradoxically, few drugs are efficient to induce tobacco cessation and to decrease relapse. Until a few weeks ago nicotin replacement therapy and bupropion only were validated in this indication. Varenicline (Champix), a partial agonist of nicotinic receptors, is a novel type of drugs currently also accepted for sale for smoking cessation. This article dwells on recent publications about efficacy of varenicline. Position, side-effects and advantages of the drug are discussed. PMID- 17461304 TI - Surgery in scandinavia. PMID- 17461305 TI - Emergency thoracotomy in trauma: rationale, risks, and realities. AB - Emergency department thoracotomy (EDT) may serve as a life-saving tool when performed for the right indications, in selected patients, and in the hands of a trained surgeon. Critically injured patients 'in extremis' arrive at an increasing rate in the trauma bay, as an effect of improved pre-hospital trauma systems and rapid transport. Any patient in near, or full cardiovascular shock prompts the trauma surgeon to rapidly perform a thoracotomy. The EDT procedure is managed best by surgeons familiar with, and experienced in, penetrating cardiothoracic injuries. However, the geographical differences in trauma epidemiology lends no, or only scarce, experience with this procedure in most European trauma centres. Consequently, mandatory training is imperative for success. The rationale for performing an EDT is to: (I) resuscitate the agonal patient with penetrating cardiothoracic injuries; (II) release cardiac tamponade by evacuation of pericardial blood; (III) immediately control hemorrhage and repair cardiac or pulmonary injury; (IV) perform open cardiac massage; and (V) place a thoracic aortic cross-clamp to redistribute the remaining blood volume, and perfuse the carotids and coronary arteries. The prevalence rates of blood borne viruses reported in critically injured patients in the USA (10-20%) exceed the prevalence in the Nordic countries (HIV prevalence < 1% in general population). However, risk is not negligible and mandated universal precautions are needed. The literature is rich in series describing the use of EDT, however, the best evidence is derived from a few prospective trials. EDT saves about one in every five patients with isolated penetrating cardiac injury, while > 98% die after blunt injury. Based on an updated review of the current available literature, this paper presents the current evidence regarding the rationale, risk, and outcomes for employing EDT in the field of trauma surgery. PMID- 17461306 TI - Role of infrainguinal angioplasty in the treatment of critical limb ischaemia. AB - OBJECTIVE: To review the published papers reporting on the use of infrainguinal angioplasty in the treatment of critical limb ischaemia (CLI). METHODS: A MEDLINE (1966-2005) and Cochrane library search for articles relating to the use of infrainguinal angioplasty in the treatment of CLI. RESULTS: Recent papers reporting on the results of infrainguinal angioplasty as treatment for CLI patients show excellent limb salvage rates regardless of the patency rates. The Cochrane Database of systematic reviews has accepted two prospective randomised trials comparing bypass operations and angioplasty among CLI patients. Pooling both trials showed no overall significant difference in amputation rates between the surgery and PTA groups. A multicentre, randomised controlled trial, the BASIL (Bypass versus Angioplasty in Severe Ischaemia of the Leg) trial showed that in the medium term (after six months), the outcomes after angioplasty or surgery among CLI patients did not differ significantly with respect to amputation-free survival, all-cause mortality and quality of life. CONCLUSIONS: Infrainguinal PTA is feasible in CLI patients. Data from the BASIL trial show the similar ability of bypass surgery and balloon angioplasty in preserving both life and limb in short term. These results are, however, not applicable for the majority of CLI patients as only 15% (70/456) of the patients with severe limb ischaemia were considered candidates for the trial. PMID- 17461307 TI - Surgical critical care: from old boundaries to new frontiers. AB - The surgical specialty of critical care has evolved into a field where the surgeon manages complex medical and surgical problems in critically ill patients. As a specialty, surgical critical care began when acutely ill surgical patients were placed in a designated area within a hospital to facilitate the delivery of medical care. As technology evolved to allow for development of increasingly intricate and sophisticated adjuncts to care, there has been recognition of the importance of physician availability and continuity of care as key factors in improving patient outcomes. Guidelines and protocols have been established to ensure quality improvement and are essential to licensing by state and national agencies. The modern ICU team provides continuous daily care to the patient in close communication with the primary operating physician. While the ultimate responsibility befalls the primary physician who performed the preoperative evaluation and operative procedure, the intensivist is expected to establish and enforce protocols, guidelines and patient care pathways for the critical care unit. It is difficult to imagine modern surgical ICU care without the surgical critical care specialist at the helm. PMID- 17461308 TI - Operation for primary hyperparathyroidism: the new versus the old order. A randomised controlled trial of preoperative localisation. AB - BACKGROUND AND AIMS: In patients with primary hyperparathyroidism (PHPT), parathyroid imaging is nowadays routinely used for the purpose to perform a focused unilateral minimally invasive operation. The outcome of this new strategy has, however, not been established in randomised trials. MATERIAL AND METHODS: Patients were randomised to either preoperative localisation with sestamibi scintigraphy and ultrasonography (group I) or no preoperative localisation (group II). In group I, a minimally invasive parathyroidectomy was performed in patients in whom both localisation studies were consistent with a single pathological gland, whereas a conventional bilateral neck exploration was performed in cases with negative localisation findings. In group II all patients underwent conventional bilateral neck exploration. Primary outcome measure was normocalcaemia at 6 months postoperatively. RESULTS: In the preoperative localisation group (group I) 23/50 (46%) of the patients could be operated on with the focused operation whereas 26/50 (52%) were operated on by bilateral neck exploration. All patients in the no localisation group (group II; n = 50) were operated on with the intended bilateral neck operation. Normocalcaemia was obtained in 96% and 94% in group I and II, respectively. Total (localisation and operative) costs were 21% higher in group I. CONCLUSIONS: Routine preoperative localisation, with the intention to perform minimally invasive parathyroidectomy, is not cost effective if concordant results of scintigraphy and ultrasonography are a prerequisite for the focused operation. Less than half of the patients were successfully managed with this strategy, at a higher cost and without obtaining a more favourable clinical outcome. PMID- 17461309 TI - Ligasure vessel sealing system versus conventional vessel ligation in thyroidectomy. AB - BACKGROUND AND AIMS: The Ligasure Vessel Sealing System (LVSS) is a new surgical technology which has been used to secure haemostasis in various fields of surgery. There is little information in the literature about the use of the LVSS in thyroid surgery and the results of these studies are controversial. The aim of this study was to report our experience with the LVSS in thyroid surgery and to compare it with our experience with conventional vessel ligation. PATIENTS AND METHODS: The non-randomized retrospective review included 403 consecutive patients who underwent primary thyroid surgery. A conventional suture-ligation technique was used in 199 patients (conventional group) and the LVSS was used in 204 patients (ligasure group). The following data were collected: patients' demographics, thyroid pathology, type of thyroidectomy, operating time and complications. RESULTS: The mean operating time for lobectomy, subtotal thyroidectomy and total thyroidectomy was significantly shorter in the Ligasure group compared with the conventional group. Average saving in operating time in the Ligasure group was 25.8 minutes (95% CI 19.5-32.2; p < 0.0001) with relative reduction being 26.6%. Among all types of complications only postoperative transient hypoparathyroidism demonstrated a significantly lower incidence in the Ligasure group (2.5% vs 7.0%; p = 0.022). CONCLUSIONS: We found that the LVSS is a safe method with minimal complications in thyroid surgery. Use of the LVSS for all types of thyroidectomy ensures a significantly shorter operating time. PMID- 17461310 TI - Clinical outcome after D1 vs D2-3 gastrectomy for treatment of gastric cancer. AB - BACKGROUND: Clinical benefit from extended lymphadenectomy for gastric cancer remains controversial as a considerable variation exists between results of different studies. METHODS: 562 patients were treated at HUCH between 1987-2003, whereof 223 underwent gastrectomy with curative intent. Of these, 114 patients underwent subtotal/total gastrectomy with D1 (standard) lymphadenectomy and 109 patients had D2-3 (extended) lymph node dissection. The clinical outcome of these patients was analysed retrospectively. RESULTS: The incidence of surgical complications was 33.0% in D2-3 and 16.8% in D1 lymphadenectomy groups (p = 0.008). Abscess was the most common complication (11.0%) among D2-3 operated patients and haemorrhage (4.4%) in D1 group. Hospital mortality was 3.7% in D2-3 and 1.8% in D1 group (p = 0.438). The only statistically significant factor influencing the rate of complications was D2-3 lymphadenectomy (OR 2.620, 95% C.I. 1.375 to 4.991). D2-3 was associated with a longer postoperative hospital stay and operation time, greater blood loss and increased need for blood transfusions compared to D1. The 5-year survival was not statistically different between lymphadenectomy groups. CONCLUSION: It is justified to perform a D2-3 gastrectomy in Europe with a acceptable postoperative mortality but with a significant morbidity. Further studies are needed to assess the value of extended lymphadenectomy in gastric cancer. PMID- 17461311 TI - Organisation and early outcomes of major upper gastrointestinal cancer surgery in Denmark 1996-2004. AB - BACKGROUND: To assess the relationship between hospital volume and early postoperative outcome the incidence and early outcome of all esophagectomies, pancreaticoduodenectomies and gastric resections in Denmark from 1996 to 2004 was described. METHODS: The National Patient Registry and discharge information from all hospital departments were analysed for all the operations due to a malignant diagnosis. All information was examined for postoperative length of stay and hospital mortality. RESULTS: During the study period 26 departments performed at least one esophageal resection, 13 departments performed at least one Whipple procedure and 37 departments performed at least one gastric resection. Four departments performed more than 20 esophageal resections per year, whereas one department performed more than 20 Whipple procedures and one more than 20 gastric resections per year. The overall mean length of stay was 21.6 days, 24 days and 18 days for esophageal, pancreatic and gastric resections, respectively, with no difference between high and low volume departments. The hospital mortality was 8.6%, 8.9% and 8.2%, respectively. CONCLUSION: The overall high mortality and long postoperative stay in patients undergoing upper gastrointestinal cancer surgery in Denmark calls for improvement by regionalisation into 3-4 departments and monitoring of results. PMID- 17461312 TI - Immunonutrition in elective gastrointestinal surgery patients. AB - BACKGROUND: Previous trials have shown that perioperative immunonutrition could protect patients from infectious complications after gastrointestinal cancer operations. The purpose of this study was to determine whether perioperative immunonutrition decreases postoperative morbidity, especially infection complications, mortality and length of hospital stay in patients undergoing major gastrointestinal tract surgery. METHODS: One hundred patients with a planned elective operation for benign or malignant gastrointestinal illness were randomized into two groups: group 1) oral supplementation for five days before and five days after surgery with 900 mL/day of a formula enriched with arginine, gamma-3-fatty acid and RNA + liquid diet ad libitum on one and two postoperative day and then solid food (immunonutrition group; n = 50) or group 2) no artificial nutrition before and after surgery, on one and two postoperative day intravenous solution of 5% glucose and electrolytes and then normal diet (conventional group; n = 50). RESULTS: The groups were comparable for all key baseline and surgical characteristics. There were nine (18%) infectious complications in both groups. Overall complication rates were 28% (n = 14) in the immunonutrition group and 24% (n = 12) in the conventional group. No significant difference between the groups was found in complication rates, mortality or length of hospital stay. CONCLUSION: Routine perioperative immunonutrition to the patients undergoing major gastrointestinal surgery is not beneficial. PMID- 17461313 TI - Intraoperative ultrasonography in detecting and assessment of colorectal liver metastases. AB - OBJECTIVE: to assess the clinical value of intraoperative ultrasonography (IOUS) in detecting and assessment of liver metastatic tumours in colorectal cancer patients. METHODS: a study is a retrospective analysis of 388 patients operated on for colorectal carcinoma between 1997 and 2004. In all the patients intraoperative ultrasound was performed. The authors analyzed of sensitivity, specificity, PPV, NPV and accuracy of pre- and intraoperative ultrasonography in detecting and staging of colorectal metastatic lesions. RESULTS: Intraoperative ultrasonography showed the highest sensitivity, specificity and accuracy in both, tumor detection (99.1, 98.5 and 98.9%, respectively) and assessment (95.4, 99.5 and 99.1%, respectively). Overall sensitivity of IOUS was significantly better in detection and staging compared with preoperative ultrasonography 91.1 and 72.2%, respectively). CONCLUSIONS: IOUS should be used as routine diagnostic modality in colorectal cancer patients with hepatic metastases or suspected metastases. Transabdominal ultrasonography cannot be used as the only diagnostic tool in the evaluation of liver lesions, but may be helpful in preoperative screening. PMID- 17461314 TI - Resection rectopexy for external rectal prolapse reduces constipation and anal incontinence. AB - BACKGROUND AND AIMS: The main aim was to examine constipation and anal incontinence in patients before and after resection for external rectal prolapse. MATERIAL AND METHODS: Twenty patients had ligament preserving suture rectopexy and sigmoid resection (resection rectopexy) for external rectal prolapse by laparoscopic (n = 15) or open (n = 5) technique during 2001-2005. They were prospectively evaluated for constipation and anal incontinence using validated incontinence and KESS-constipation scores. RESULTS AND CONCLUSIONS: Constipation score was significantly reduced from mean 7.7 (5.4-9.9) to 4.5 (2.5-6.4) after median 4 months (1-19) and to 4.3 (2.2-6.3) after median 17 months (4-51). Six and four patients were constipated preoperatively and 17 months postoperatively, respectively. The four symptoms feeling incomplete evacuation of stool, minutes in lavatory per attempt, use of enemas/digitation and painful evacuation effort were significantly reduced, whilst stool consistency increased. Fourteen patients (70%) had anal incontinence. Corresponding and significant reduction in their scores were from mean 12.5 (9.4-15.5) to 5.1 (2.1-8.1) and to 3.6 (1.3-5.9). Incontinence was improved in 13 and unaltered in one patient(s). Two patients with worse outcome had increased stool consistency and constipation scores. Resection rectopexy for rectal prolapse reduced anal incontinence and constipation. PMID- 17461315 TI - A 19-year review of paediatric patients with acute scrotum. AB - BACKGROUND AND AIMS: The aim of the study was to compare incidence, symptoms and signs of spermatic cord torsion to those of other conditions causing acute scrotum. MATERIAL AND METHODS: Records of 388 consecutive boys under 17 years of age treated for acute scrotum at The Hospital for Children and Adolescents in Helsinki in 1977-1995 were reviewed. During the period studied all patients with acute scrotum underwent urgent surgery to ensure accurate diagnosis and treatment. The duration and characteristics of the symptoms, clinical findings prior to operation and the age of the patients were registered. RESULTS: Scrotal explorations revealed 100 cases (26%) of spermatic cord torsion (SCT), 174 cases (45%) of torsion of the testicular appendage (AT), 38 cases (10%) of epididymitis (ED), 32 cases (8%) of incarcerated inquinal hernias and 44 (11%) other conditions. During the first year of life SCT was the most common cause of acute scrotum, another peak incidence being in adolescence. Almost half of the boys with AT were nine to 12 years of age (median 11). Except for infants, the patients' acute symptoms were pain (SCT 88%, AT 94%, ED 76%). Swelling in the hemiscrotum was found in 44% of SCT, in 39% of AT and in 88% of ED cases. Epididymitis was also accompanied by erythema (37%), but infrequently with fever (in 16%). Erythema was found also in AT (32%), but the "blue dot sign" was found positive in only 17 (10%) of the boys with AT. Three quarters of the boys who were operated on within six hours from onset of symptoms had testicle torsion. All testicles were saved when detorsion was performed within six hours, but salvage was possible in only half of the cases when symptoms had lasted more than six but less than 12 hours. CONCLUSIONS: The high probability of SCT among those admitted to an emergency department within six hours from the onset of the symptoms justifies immediate surgical exploration. PMID- 17461316 TI - Emergency room arteriography: an updated digital technology. AB - BACKGROUND: Emergency room arteriography (ERA) is a safe, accurate, simple and cost-effective method of defining arterial injuries. Limitations include the difficulty of evaluating limb vasculature distal to the suspected site of injury. Statscan is a novel, low-dose digital X-ray machine that can rapidly obtain a whole body image in a single scan. Our goal was to evaluate the role of Statscan technology in ERA. METHODS: A 24 month retrospective review of all patients who underwent a Statscan assisted ERA at the Groote Schuur Hospital Trauma Unit was completed. Indications for ERA included a hemodynamically stable patient with hard signs of a vascular injury in conjunction with the clinical assessment of a threatened limb. Contraindications encompassed instability, massive bleeding or a rapidly expanding hematoma. RESULTS: Ten patients underwent Statscan assisted ERA of their lower limbs. Eight had cold, pulseless limbs with impaired neurological examinations. Common femoral, superficial femoral and popliteal artery lacerations were displayed. Three patients had no identifiable injury and were observed. Seven patients underwent operative management for threatened limbs. Two had Statscan evidence of arterial emboli distal to the site of injury leading to further exploration and distal embolectomy. CONCLUSIONS: Statscan ERA is safe, rapid, simple and accurate. It has the advantage of providing arteriography distal to the site of injury. This directly altered patient care in 20% of cases, primarily by detecting distal arterial emboli. Thirty percent of patients with normal ERA also avoided an unnecessary operation. This study demonstrates a new role for Statscan technology. PMID- 17461317 TI - A comparison of the haemodynamic effects of 4% succinylated gelatin, 6% hydroxyethyl starch (200/0.5) and 4% human albumin after cardiac surgery. AB - BACKGROUND AND AIMS: The goal for volume replacement therapy is to maintain stable haemodynamics after cardiac surgery. We hypothesized that a short term infusion of hydroxyethyl starch results in better haemodynamic response than an infusion of lower molecular weight gelatin. MATERIAL AND METHODS: 45 patients received a predetermined fixed dose of 15 ml kg(-1) of either 4% succinylated gelatin (GEL) or 6% hydroxyethyl starch (HES) or 4% human albumin (HA) after cardiac surgery. RESULTS AND CONCLUSIONS: Pulmonary capillary wedge pressure was more increased in GEL and HES groups [mean (SD) 153% (54) and 168% (57) of pre infusion value] than in HA group [122% (23)] (P = 0.031) after completion of infusion, but no differences in cardiac index (CI) and stroke volume index (SVI) were observed. At 2 and 18 hours after end of study infusions SVI was more increased in HES [143% (38) and 148% (41) of pre-infusion values] and HA [143% (35) and 163% (42) of pre-infusion values] groups than in GEL [116% (23) and 125% (30)] group (P = 0.047 at 2 hours and P = 0.033 at 18 hours). In early postoperative phase after cardiac surgery, HES and HA infusions improve haemodynamics more and longer period than GEL infusion. PMID- 17461318 TI - Traumatic injuries associated with horseback riding. AB - BACKGROUND: According to the Center for Disease Control and Prevention (CDC), an estimated 30 million people ride horses each year in the United States. Horseback riding related injuries are common, with an estimated 50,000 emergency room visits annually. The popularity of recreational horseback riding has increased in South Florida and the incidence of associated traumatic injuries is a reflection of this. MATERIAL AND METHODS: Retrospective review of patients admitted to a state designated Level I trauma center that sustained horseback riding associated injuries between January 2000 and December 2003. Information extracted from the Trauma Center's data base included demographics, mechanism of injury and toxicology screening. RESULTS: During the review period, twenty-seven patients were identified. There were 12 men and 15 women. The average age was 36 years. The injuries occurred during pleasure riding in 23 patients and thoroughbred related activities in 4 patients. Multiple severe injuries were common and documented in 24 patients. All patients required hospitalization with an average stay of 5 days. Five patients had a positive toxicology screen on admission. No deaths were documented in this review. CONCLUSION: Horseback riding related injuries tends to be serious. Alcohol and recreational drugs may contribute to exacerbate the extent of these injuries. The use of proper protective equipment, instructions for safe riding, and discouraging drug and alcohol use during riding activities should be emphasized. PMID- 17461319 TI - Arthroscopic anterior cruciate ligament reconstruction using a hamstring graft with interference screw fixation. AB - BACKGROUND AND AIMS: This study assessed the clinical and radiographic results 3 to 8 years after an arthroscopic ACL reconstruction using a hamstring graft with interference screw fixation on femoral and tibial sites. MATERIAL AND METHODS: Seventy-six of 100 patients, who underwent an arthroscopic ACL reconstruction, could be re-examined with a mean follow-up of 5 years. RESULTS: The mean Tegner activity score was 4 (scale: 0 to 10) before surgery, and 5 at the follow-up. Fifty-four patients (78%) achieved a satisfactory result in the IKDC rating. The mean Lysholm score was 85 (scale: 0 to 100), the score being better in patients without meniscal resection than those with (P = 0.04). The mean side-to-side difference in anterior-posterior knee laxity was 1.8 mm, and 62 patients (90%) had good stability. Sixty-four patients (93%) had normal range of motion of the knee. Twelve patients (46%) with meniscal resection had osteoarthritic changes, compared with only 8 of those (20%) without meniscal resection (P = 0.03). CONCLUSIONS: This medium-term follow-up study showed that good stability and function and normal range of motion of the knee can be achieved in majority of the patients who have undergone an ACL reconstruction with a hamstring graft and interference screw fixation. PMID- 17461320 TI - Seroprevalence of bloodborne viruses in Scandinavian trauma victims. PMID- 17461321 TI - Long acting beta2 agonists: are they safe? PMID- 17461322 TI - Biliary tract injury during cholecystectomy: a retrospective descriptive review of clinical features, treatment and outcome. PMID- 17461323 TI - An outbreak of Aspergillus meningitis following spinal anaesthesia for caesarean section in Sri Lanka: a post-tsunami effect? AB - An outbreak of Aspergillus fumigatus meningitis occurred in 5 women following spinal anaesthesia, performed between 21 June and 17 July 2005 for caesarean section, in Colombo, Sri Lanka. The patients' median age was 27 years. Different teams in 2 maternity hospitals gave spinal anaesthesia. Mean incubation period was 11.2 days. Fever, headache and nuchal rigidity were common presentations. Remittent fever continued despite broad-spectrum intravenous antibiotics. Papilloedema, lateral rectus palsy, cerebral infarction and haemorrhage developed later. Three patients died. Cerebrospinal fluid pleocytosis with low glucose yielded negative PCR for fungi. Fungal cultures subsequently grew Aspergillus fumigatus. A post-mortem of the first patient confirmed Aspergillus meningitis, followed by treatment with amphotericin B and voriconazole, that saved the lives of others. Visual and hearing impairment in one and complete recovery in the other were observed a year after treatment. Examination of unused plastic syringes, needles, cannulae, and ampoules of anaesthetic agents confirmed that 43 syringes from three different manufactures were contaminated with Aspergillus fumigatus. The stores for drugs and devices of the Ministry of Health were examined and found to be full of tsunami donations, while regular procurements of the Ministry were kept in a poorly maintained humid warehouse. Inadequate space for tsunami donations was identified as the most plausible explanation for sub optimal storage. Withdrawal and incineration of all unused syringes controlled the outbreak. The survival of those aggressively treated for Aspergillus meningitis suggests in hindsight that the availability of diagnostic tests and specific treatment, and early recognition of the outbreak could have saved the lives of victims who died. Early life-threatening side-effects and permanent long term sequelae of antifungal medication stress the need to be cautious with empirical treatment in immuno-competent low-risk individuals. PMID- 17461324 TI - Western model of community mental health care: its applicability to Sri Lanka. PMID- 17461325 TI - Subcutaneous local recurrence of parathyroid carcinoma. PMID- 17461326 TI - A rare cervical spine abnormality associated with neurofibromatosis. AB - Spinal abnormalities such as kyphoscoliosis and vertebral scalloping are frequent occurrences in type 1 neurofibromatosis (NF1). We report this rare case of posterior displacement of C3 vertebral body into the spinal canal causing severe cord compression associated with neurofibromatosis in a 13-year old girl. She underwent anterior excision of C3 vertebral body, bone graft and fixation of C2- C4, with improvement of her symptoms. PMID- 17461327 TI - A case of hyper-IgE syndrome. AB - Hyper-IgE syndrome, a multi-system disorder affecting dentition, skeletal and immune systems and connective tissues, presents with recurrent infections and dermatitis. We report here the first case in Sri Lanka. PMID- 17461328 TI - Trap-gun injuries--a menace in rural agricultural areas. PMID- 17461329 TI - Surgical complications after renal transplantation. PMID- 17461330 TI - Objective assessment of outcome of transinguinal varicocele ligation in oligozoospermic subfertile men. PMID- 17461331 TI - Histopathology reporting in colorectal cancer. PMID- 17461332 TI - Filarial antigens detected in urine using the immunochromatographic card test. PMID- 17461333 TI - Condoms and the Vatican. PMID- 17461334 TI - Social networks and HIV/AIDs risk perceptions. AB - Understanding the determinants of individuals' perceptions of their risk of becoming infected with HIV and their perceptions of acceptable strategies of prevention is an essential step toward curtailing the spread of this disease. We focus in this article on learning and decision-making about AIDS in the context of high uncertainty about the disease and appropriate behavioral responses. We argue that social interactions are important for both. Using longitudinal survey data from rural Kenya and Malawi, we test this hypothesis. We investigate whether social interactions--and especially the extent to which social network partners perceive themselves to be at risk--exert causal influences on respondents' risk perceptions and on one approach to prevention, spousal communication about the threat of AIDS to the couple and their children. The study explicitly allows for the possibility that important characteristics, such as unobserved preferences or community characteristics, determine not only the outcomes of interest but also the size and composition of networks. The most important empirical result is that social networks have significant and substantial effects on risk perceptions and the adoption of new behaviors even after we control for unobserved factors. PMID- 17461335 TI - Orphans and schooling in Africa: a longitudinal analysis. AB - AIDS deaths could have a major impact on economic development by affecting the human capital accumulation of the next generation. We estimate the impact of parent death on primary school participation using an unusual five-year panel data set of over 20,000 Kenyan children. There is a substantial decrease in school participation following a parent death and a smaller drop before the death (presumably due to pre-death morbidity). Estimated impacts are smaller in specifications without individual fixed effects, suggesting that estimates based on cross-sectional data are biased toward zero. Effects are largest for children whose mothers died and, in a novel finding, for those with low baseline academic performance. PMID- 17461336 TI - Educational reversals and first-birth timing in sub-Saharan Africa: a dynamic multilevel approach. AB - In many areas throughout sub-Saharan Africa, young adult cohorts are less educated than their predecessors because of declines in school enrollments during the 1980s and 1990s. Because a woman with little education typically becomes a mother earlier and has more children than one with better education, and because of a similar well-established relationship between current education and current fertility at the societal level, one might expect such education reversals to raise fertility. However, if there is an additional negative effect of low educational level among currently young women compared with that in the past, which would accord with ideas about the impact of relative deprivation, the total effect of an education reversal may run in either direction. This possibility has not been explored in earlier studies, which have taken a more static approach. We focus on the initiation of childbearing. Using Demographic and Health Survey data from 16 sub-Saharan African countries with multiple surveys, we estimate a fixed effects multilevel model for first births that includes the woman's own education, community education, and community education relative to the past. There are negative effects of individual and community education, but no effect of relative education. Thus we conclude that education reversals do seem to speed up entry into parenthood. PMID- 17461337 TI - Child gender and father involvement in fragile families. AB - In this article, we use data from the first two waves of the Fragile Families and Child Wellbeing Study to examine the effects of child gender on father involvement and to determine if gender effects differ by parents' marital status. We examine several indicators of father involvement, including whether the father acknowledges "ownership" of the child, whether the parents live together when the child is one year old, and whether the father provides financial support when the child is one year old. We find some evidence that child gender is associated with unmarried father involvement around the time of the child's birth: sons born to unmarried parents are more likely than daughters to receive the father's surname, especially if the mother has no other children. However, one year after birth, we find very little evidence that child gender is related to parents' living arrangements or the amount of time or money fathers invest in their children. In contrast, and consistent with previous research, fathers who are married when their child is born are more likely to live with a son than with a daughter one year after birth. This pattern supports an interpretation of child gender effects based on parental beliefs about the importance of fathers for the long-term development of sons. PMID- 17461338 TI - Child support and father-child contact: testing reciprocal pathways. AB - I use three waves of panel data to examine the relationship between child support payments and fathers' contact with their nonmarital children. I disaggregate support into fathers' formal and informal payments and incorporate cross-lagged effects models to identify the direction of causality between payments and contact. After including the behavior from the prior wave (lagged term) and a rich set of family characteristics, I find a marginally significant effect of paying formally at Time 1 on the likelihood of contact at Time 2 but no effect of contact at Time 1 on formal payments at Time 2. In the first examination of the relationship between informal support and father-child contact, I find a strong, positive reciprocal relationship between the likelihood and frequency of father child contact and the likelihood and amount of informal support, with slightly stronger and more consistent effects of contact on payments than of payments on contact. PMID- 17461339 TI - The long-term effects of poor childhood health: an assessment and application of retrospective reports. AB - This study assesses retrospective childhood health reports and examines childhood health as a predictor of adult health. The results suggest that such reports are of reasonable reliability as to warrant their judicious use in population research. They also demonstrate a large positive relationship between childhood and adult health. Compared with excellent, very good, or good childhood health, poor childhood health is associated with more than three times greater odds of having poor adult self-rated health and twice the risk of a work-limiting disability or a chronic health condition. These associations are independent of childhood and current socioeconomic position and health-related risk behaviors. PMID- 17461340 TI - Gender differences in the link between childhood socioeconomic conditions and heart attack risk in adulthood. AB - A growing body of evidence shows that childhood socioeconomic status (SES) is predictive of disease risk in later life, with those from the most disadvantaged backgrounds more likely to experience poor adult-health outcomes. Most of these studies, however are based on middle-aged male populations and pay insufficient attention to the pathways between childhood risks and specific adult disorders. This article examines gender differences in the link between childhood SES and heart attack risk trajectories and the mechanisms by which early environments affect future disease risk. By using methods that model both latent and path specific influences, we identify heterogeneity in early life conditions and human, social, and health capital in adulthood that contribute to diverse heart attack risk trajectories between and among men and women as they age into their 60s and 70s. We find that key risk factors for heart attack operate differently for men and women. For men, childhood SES does not differentiate those at low, increasing, and high risk for heart attack. In contrast, women who grew up without a father and/or under adverse economic conditions are the most likely to experience elevated risk for heart attack, even after we adjust for the unequal distribution of working and living conditions, social relationships, access to health care, and adult lifestyle behaviors that influence health outcomes. PMID- 17461341 TI - Income dynamics in couples and the dissolution of marriage and cohabitation. AB - Several studies have shown that a wife's strong (socio)economic position is associated with an increase in the risk of divorce. Less is known about such effects for cohabiting relationships. Using a unique and large-scale sample of administrative records from The Netherlands, we analyze the link between couples' income dynamics and union dissolution for married and cohabiting unions over a 10 year period. We find negative effects of household income on separation and positive effects of the woman's relative income, in line with earlier studies. The shape of the effect of the woman's relative income, however, depends on the type of union. Movements away from income equality toward a male-dominant pattern tend to increase the dissolution risk for cohabiting couples, whereas they reduce the dissolution risk for married couples. Movements away from income equality toward a female-dominant pattern (reverse specialization) increase the dissolution risks for both marriage and cohabitation. The findings suggest that equality is more protective for cohabitation, whereas specialization is more protective for marriage, although only when it fits a traditional pattern. Finally, we find that the stabilizing effects of income equality are more pronounced early in the marriage and that income equality also reduces the dissolution risk for same-sex couples. PMID- 17461342 TI - Should I stay or should I go? The impact of age norms on leaving home. AB - This article studies the association between social norms and the timing of leaving home. Although largely overlooked by most recent studies on leaving home, life-course theory suggests that age norms and age grading influence life-course decisions in general and leaving home in particular. We use Fishbein and Ajzen's model of "reasoned behavior" to integrate this strand of research with the more individualistic view that dominates current thinking. Using data from a Dutch panel survey, we use a Cox regression model with a control for sample selection to estimate the association between perceived age norms and the timing of leaving home. We show that perceived opinions of parents are associated with the actual timing of leaving the parental home but that societal norms and friends' norms concerning the timing of leaving home are not. In addition, the timing of leaving home is also associated with the perceived costs and benefits of leaving home and with the perceived housing market situation. PMID- 17461343 TI - [Mandibular movements in speech: interference of temporomandibular disfunction according to pain indexes]. AB - BACKGROUND: temporomandibular disorders can cause general alterations of the mandibular movements due to modification in the condition of muscles and articulations. Electrognathography, a computerized exam used to complement the diagnosis of these disorders, allows the objective delineation and record of the mandibular movements, determining their amplitude and speed. AIM: to verify the characteristics of mandibular movements of individuals with temporomandibular dysfunction and in asymptomatic individuals during speech, through computerized electrognathography, analyzing possible interferences of this dysfunction and severity implications regarding pain indexes. METHOD: 135 adults were divided in four groups based on their pain indexes, using a numeric scale: zero for pain absence, one for mild pain, two for moderate pain and three for severe pain. Mandibular movements were observed during the sequential naming of balanced pictures taking in consideration the occurrence of phonemes in the Brazilian Portuguese language. Records were obtained using computerized electrognathography (BioEGN - BioPak system). RESULTS: the analysis of the results point that differences indicated as significant for mandibular opening amplitude and mandibular closing speed occurred between index zero and all of the other pain indexes. Regarding mandibular opening speed during speech, statistically significant differences were obtained between index zero and index three. It was observed that mandibular movements in speech are discreet, with an anteroposterior component and deviations in laterality. CONCLUSION: the presence of temporomandibular dysfunctions causes reduction in the values of maximum mandibular opening and a reduction in both mandibular opening speed and mandibular closing speed during speech The different pain indexes: mild, moderate and severe do not seem to determine larger reduction of these values. PMID- 17461344 TI - [Vocal perception and life quality]. AB - BACKGROUND: the perception of dysphonia and the impact of vocal alteration on life quality. AIM: to verify if the interference of dysphonia on life quality is related to the vocal self-perception of dysphonic individuals and to the perception members of the community have about the vocal pleasantness of these individuals. METHODS: 31 dysphonic adults, prior to treatment, filled out the Voice Related Quality of Life (V-RQOL) protocol that analyzes the impact of dysphonia on life quality. These individuals were also asked to comment about their self-perception of vocal quality. Recordings of the sustained "a" vowel and number counting from one to ten were made, using a digital system in an acoustic treated environment. The recorded material was presented to 25 judges, with no training in vocal perception, so that they could classify the voices according to the same scale used by the dysphonic individuals when commenting about their own voices. RESULTS: according to Spearman's test, a statistically significant correlation was observed between the results of the V-RQOL and the vocal self perception related to the social/emotional (p = 0.047), physical (0 = 0.010) and total (p = 0.008) domains. However there was no correlation between the results of the V-RQOL and the perception of the judges, both for the sustained "a" vowel and for the number counting (p = 0.475 e p = 0.152 respectively). CONCLUSION: it was observed that the worst the opinion of the dysphonic individual about the impact of dysphonia on life quality, the worst was his vocal self-perception. A relationship between the life quality of the dysphonic individual and the vocal perception of listeners was not observed. PMID- 17461345 TI - [Sensory oral motor and global motor development of preterm infants]. AB - BACKGROUND: development assessment of preterm infants. AIM: to evaluate the association between the gestational ages (GA) of premature infants with the global motor development as well as with early signs of sensory oral motor development delay, and to verify a possible association between them. METHOD: an exploratory study that assessed the development of 55 infants with corrected chronological ages between four to five months, born preterm at the Instituto Materno Infantil Professor Fernando Figueira (IMIP) and who were followed at the Kangaroo Mother Program Clinic between March and August of 2004. The assessment of the sensory oral motor development was performed through pre-selected indicators and of the global motor development through the Alberta Infant Motor Scale (AIMS). RESULTS: infants with lower GA (29 to 34 weeks) presented a higher median of risk signs in the sensory oral motor development assessment when compared to those with higher GA (35 to 36 weeks). Regarding the global motor development, infants born with lower GA presented a higher number of scores in the AIMS below percentile 10 (26%) when compared to those with a higher GA (4%) (p=0.009). The median index of the risk signs for the sensory oral motor development were significantly higher among infants with total AIMS scores below percentile 25 when compared to those with scores equal to or above percentile 25. CONCLUSION: the gestational age of infants at birth influenced the sensory oral motor and global motor development - infants with lower gestational ages presented worse performances. These findings suggest a possible association between both aspects of infant development. PMID- 17461346 TI - [Study of benefit and of acclimatization in recent users of hearing aids]. AB - BACKGROUND: benefit and acclimatization. AIM: to characterize the short-term benefits presented by adults, recent users of hearing aids, through objective (functional gain) and subjective (self-evaluation questionnaires) procedures. The possible occurrence of the phenomenon of acclimatization was also investigated based on the analysis of the speech recognition percent index (SRPI) before the fitting of the hearing aids and after four and 16/18 weeks of hearing aids use. METHOD: sixteen subjects, with ages between 17 and 89 years, with symmetric moderate or severe sensorineural hearing loss, recent hearing aid users, were evaluated. The study was divided in three phases: before the hearing aids fitting, and after four and 16/18 weeks following the fitting process. In all of the phases the following procedures were used: pure tone audiometry, speech recognition percent index, speech recognition threshold and the Hearing Handicap Inventory for Elderly Screening Version or the Hearing Handicap Inventory for the Adults Screening Version, and the Abbreviated Profile of Hearing Aid Benefit. After the hearing aid fitting process, subjects were submitted to free field audiometry. RESULTS: results showed statistically significant differences between objective and subjective measures after the use of hearing aids, indicating short term benefit. However, as time went by, the benefit obtained with the use of hearing aids did not improve significantly, suggesting that benefit does not increase with time. SRPI as well as subjective measures improved with long-term hearing aid use, but such differences were not statistically significant. CONCLUSIONS: short-term benefits were obtained with the use of hearing aids, however the phenomenon of acclimatization was not observed through the SRPI. PMID- 17461347 TI - [Analysis of the pragmatic abilities profile in normal preschool children]. AB - BACKGROUND: pragmatic abilities in children. AIM: to analyze the pragmatic abilities profile in normal preschool children and to verify if significant differences exist regarding the children's different socioeconomic levels. METHOD: participants of this study were 30 children, with ages between 36 and 47 months, who attended public and private elementary schools - low and medium/high socioeconomic levels respectively. A thirty minute semi-structured conversation between each child and the evaluator was recorded in VHS. For the analysis of the pragmatic abilities profile 20 minutes of each conversation was transcribed. RESULTS: there is a prevalence of verbal over non-verbal and unintelligible utterances; simple over expansive utterances; coherent over incoherent utterances. There was a low occurrence of utterances used to start a conversation. Regarding the use of communicative functions, the informative function prevailed, even though all the others (instrumental, heuristic, naming, narrative, negative, interactive) were used by all of the children. Comparing the performance of children who attended public schools with that of children who attended private schools, statistically significant differences were observed in the number of simple and expansive verbal utterances, and in the use of the narrative function, indicating a better performance of children from private institutions. CONCLUSION: the analysis of the children's conversational abilities profile revealed that they respond/maintain the conversation instead of initiating it; their utterances are verbal, mostly coherent and simple. Regarding the communicative functions, the most prevalent was the informative function. Sociolinguistic aspects can interfere in the pragmatic abilities of children of different socioeconomic level. PMID- 17461348 TI - [Feeding disorders in children with gastro-esophageal reflux disease]. AB - BACKGROUND: feeding/eating disorders are frequent in pediatric patients and, in some cases, can be associated to an upper digestive motility disorder. Gastro esophageal reflux is, nowadays, considered a risk factor for the development of feeding/eating disorders. AIM: to verify the occurrence of feeding/eating disorders in patients with Gastro-Esophageal Reflux Disease (GERD) determined by the 24-H esophageal pH monitoring evaluation. METHOD: an analytical observational cross-sectional study of the stomathognatic system and feeding/eating behavior in healthy children and in children with GERD. RESULTS: 25 children (ages 45,68 +/- 34,22 months; mean +/- SD) with the diagnosis of GERD determined by the 24-H esophageal pH monitoring evaluation and 40 children (ages 60,65 + 36,07 months; mean + SD) randomized from their school group. The criteria for the pH monitoring were: vomiting, regurgitation, wheezing and recurrent pneumonia. There was no significant difference in age mean between groups. Children with GERD presented an significantly higher frequency (p<0.05) of feeding/eating problems (F/EP) and of oral motor-sensory disturbances (sucking, chewing and swallowing). Behavioral F/EP was present in 44% of the cases and oral motor-sensory F/EP in 80%. About 64% of the children had a history of feeding/eating complaints, 36% presented an extended feeding/eating time, 68% presented problems in the development of the oral feeding patterns and 60% presented alterations in the nasal breathing pattern. CONCLUSION: children with GERD presented a higher prevalence of behavioral and stomathognatic feeding/eating problems when compared to healthy children. PMID- 17461349 TI - [Comparative analyses of the effectiveness of three different phonological therapy models]. AB - BACKGROUND: phonological therapy in children with phonological disorders. AIM: to verify changes in the phonological system of children with phonological disorders, based on the comparison of number of acquired phonemes, number of sounds in the phonetic inventory and altered distinctive features, before and after therapy, and to verify differences between therapy models: Modified Cycles, Maximal Oppositions and ABAB--Withdrawal and Multiple Probes. METHOD: the research group was composed of 21 subjects, 15 male and 6 female, with phonological disorders, who had already completed speech-language treatment. The initial and final phonological assessments were compared taking into consideration the number of phonemes in the phonological systems, the number of sounds in the phonetic inventory and the altered distinctive features. Phonological changes that resulted from the application of the three therapeutic models were also compared. RESULTS: statistically significant differences were observed between the initial and final assessments in all of the three therapeutic models considering the number of acquired phonemes and the altered distinctive features, whereas in the phonetic inventory a statistically significant difference was observed only between the ABAB--Withdrawal and Multiple Probes and the Maximal Oppositions model. No statistical difference was observed between the therapy models. CONCLUSION: the three therapy models were effective in the treatment of children with phonological disorders, once all of the children presented improvement of their phonological system, phonetic inventory and altered distinctive features. No significant statistical difference was found between the models PMID- 17461350 TI - [Effect of sound amplification on parent's communicative modalities]. AB - BACKGROUND: auditory rehabilitation in deaf children users of sign language. AIM: to verify the effects of sound amplification on parent's communicative modalities when interacting with their deaf children. METHOD: participants were twelve deaf children, aged 50 to 80 months and their hearing parents. Children had severe or profound hearing loss in their better ear and were fitted with hearing aids in both ears. Children communicated preferably through sign language. The cause effect relation between the children's auditory skills profile (insertion gain, functional gain and The Meaningful Auditory Integration Scale--MAIS) and the communicative modalities (auditive-oral, visuo-spacial, bimodal) used by parents was analyzed. Communicative modalities were compared in two different experimental situations during a structured interaction between parents and children, i.e. when children were not fitted with their hearing aids (Situation 1) and when children were fitted with them (Situation 2). Data was analyzed using descriptive statistics. RESULTS: the profile of the deaf children's auditory skills demonstrated to be lower than 53% (unsatisfactory). Parents used predominately the bimodal modality to gain children's attention, to transmit and to end tasks. A slight positive effect of sound amplification on the communicative modalities was observed, once parents presented more turn-takings during communication when using the auditory-oral modality in Situation 2. CONCLUSION: hearing parents tend to use more turn-takings during communication in the auditory-oral modality to gain children's attention, to transmit and to end tasks, since they observe an improvement in the auditory skills of their children. PMID- 17461351 TI - [Temporal resolution: analysis in term and preterm preschoolers]. AB - BACKGROUND: auditory processing. AIM: to verify the hearing behavior of temporal resolution in children with ages from five to six years, who were born preterm, with no evidence of neurological alterations and to compare this behavior to that observed in children of the same age, who were born at term, with low risk for developmental disorders, taking into consideration the variables of: threshold detection gaps through pre-established frequency, binaural and monaural presentation, order of stimuli presentation and gender. METHOD: 70 children divided in two groups: Group 1 with 44 children who were born at term (20 female and 24 male) and Group 2 with 26 preterm children (12 female and 14 male). Children were submitted to audiologic evaluation composed of audiometry, speech response threshold, acoustic impedance test and the Random Gap Detection Test (RGDT). RESULTS: children who were born at term presented lower threshold detection gaps in the RGDT, for both monaural and binaural stimuli presentation, in all of the pre-established frequencies when compared to preterm children. This difference between the groups was statistically significant. The average threshold detection gaps of Group 1 rose according to the increase of frequency. For Group 2 statistically significant differences were not found regarding the average of threshold detection gaps, for both monaural and binaural stimuli presentation. CONCLUSION: preterm children differ from those born at term regarding the hearing behavior of temporal resolution. The RGDT can be used as a tool to evaluate the hearing process, once the early detection of alterations in temporal processes indicates the need for intervention in order to minimize or avoid future language impairments. PMID- 17461352 TI - [Brazilian version of the dizziness handicap inventory]. AB - BACKGROUND: impact of dizziness on life quality (LQ). AIM: to adapt the Dizziness Handicap Inventory (DHI) for application in the Brazilian population; to assess its reproducibility; and to describe the results obtained in patients with chronic dizziness. METHOD: The DHI was initially applied in 45 patients with chronic dizziness and with a clinical diagnosis of vestibular syndrome. The application followed the stages of translation--from English to Portuguese--and linguistic adaptation, grammatical and idiomatic equivalence review and evaluation of its intra and inter-researchers reproducibility. Reproducibility was assessed by using the Wilcoxon Test for two dependent samples, P < 0.05. The questionnaire was applied on 250 patients with chronic vestibular syndrome in order to evaluate the impact of dizziness on LQ. RESULTS: The Brazilian version of the DHI (Brazilian DHI) was well comprehended by the studied population and no statistically significant difference was found in the inter-researcher (P = 0.418) and intra-researcher (P = 0.244) reproducibility. All of the studied patients presented a loss in LQ due to dizziness. Aspects which were most affected were the physical ones, followed by, in a decreasing order, the functional and the emotional ones. Functional aspects were more compromised in older individuals. No association was verified between gender and the overall mean scores of the DHI and between gender and the mean scores of each aspect assessed by the DHI. CONCLUSION: the DHI was culturally adapted for application in the Brazilian population (Brazilian DHI). This instrument was considered to be a reliable tool to evaluate the impact of dizziness on LQ. Patients with chronic dizziness and with clinical diagnosis of vestibular syndrome presented a loss in LQ, due to this symptom. This loss was verified by the application of the Brazilian DHI. Physical aspects were the most compromised. PMID- 17461353 TI - [Perceptual and instrumental assessment of velopharyngeal function in asymptomatic submucous cleft palate]. AB - BACKGROUND: submucous cleft palate (SMCP) may or may not be associated to symptoms of velopharyngeal dysfunction (VPD). AIM: to verify whether patients with SMCP, who were considered as asymptomatic on a perceptual speech evaluation, present absence of hypernasality and adequate velopharyngeal closure on an instrumental assessment. METHOD: 22 patients with SMCP and with no VPD symptoms, of both genders, aged 6 to 46 years, underwent speech acoustic assessment (nasometry)--to determine nasalance scores--and speech aerodynamic assessment (pressure-flow technique)--to verify velopharyngeal closure. The complete agreement between the findings on the perceptual and instrumental assessments was the tested null hypothesis. RESULTS: the pressure-flow technique confirmed the perceptual findings, that is, all patients were diagnosed as having adequate velopharyngeal function on both types of assessment. Nasometry results confirmed the perceptual findings in 15 out of the 20 analyzed patients (75% of the cases). The remaining 5 patients (25%) presented nasalance scores that suggest hypernasality (nasometry). This symptom was not identified by the perceptual evaluation, yielding to the rejection of the null hypothesis. CONCLUSION: results indicate the importance of the combined use of perceptual and instrumental assessments for the diagnosis of VPD in SMCP cases. The follow-up of patients diagnosed as asymptomatic by a perceptual speech evaluation and presenting evidences of VPD on instrumental assessment (e.g. nasometry) is recommended, particularly when dealing with children who are more prone to develop symptoms with aging. PMID- 17461354 TI - [P300 in subjects with hearing loss]. AB - BACKGROUND: behavioral and electrophysiological evaluations contribute to the understanding of the hearing system and to the intervention process. AIM: to investigate the occurrence of P300 in subjects with congenital severe to profound hearing loss, according to the variables of gender, age and hearing loss level. METHOD: the design of this research is a descriptive transversal study. Twenty nine subjects, 15 male and 14 female, ranging in age from 11 to 42 years, were evaluated. Inclusion criteria were: to have at least 11 years of age and no more than 45 years; to have the diagnosis of congenital severe to profound sensorineural hearing loss; to have no other disorder; and absence of central hearing loss or any other auditory conductive disorder. The first stage consisted of an auditory behavioral and physiological evaluation, including: pure tone audiometry (air and bone conduction measures), speech audiometry, SDT (Speech Detection Threshold) and functional gain measures for the subjects using hearing aids, and immittance measures--tympanometry and acoustic reflexes thresholds; transient evoked otoacoustic emissions (TEOAE); distortion product otoacoustic emissions (DPOAE). The electrophysiological evaluation was the fourth stage of the study and included: auditory brainstem response (ABR) and late latency response (P300). RESULTS: P300 was obtained for 17 out of the 29 subjects. Mean latency and amplitude were 326.97 ms and 3.76 V, respectively. A statistical significant difference was observed for latency when considering the variable age (electrode CzA2 p < 0.003 and CzAI p < 0.02) and for amplitude when considering the variable hearing loss level (p < 0.0015). CONCLUSION: P300 can be recorded in subjects with hearing loss. PMID- 17461355 TI - [Efficacy of rehabilitation in oropharyngeal dysphagia]. AB - BACKGROUND: efficacy of rehabilitation in oropharyngeal dysphagia. In our country the practice of speech-language pathology in oropharyngeal dysphagia has increased significantly and, at this moment, deserves attention since practice needs to be based on scientific evidence. Therapeutic techniques and the outcome of rehabilitation in oropharyngeal dysphagia have been studied since the 70s, reaching its high point during the 80s and 90s. Few studies have investigated the efficacy of therapy in the rehabilitation of oropharyngeal dysphagia, the vast majority have tried to prove the effects of therapy on the dynamics of swallowing. In Brazil, the studies about oropharyngeal dysphagia have, in great part, investigated assessment procedures, and only a few have worried about rehabilitation. AIM: to present a critical analysis about the efficacy of rehabilitation in oropharyngeal dysphagia. CONCLUSION: this review of the literature indicates that non-randomized studies have compromised the results, once the casuistic of the researches are very heterogeneous--they include neurogenic and mechanical oropharyngeal dyshagia caused by different etiologies. Besides that, therapeutic programs which are used are not sufficiently described, compromising the reproduction of the methodology by other researchers. These results suggest the need for more randomized studies, which can be initially developed as case studies in order to exclude the control variables of therapy efficacy. Another suggestion is, as proposed by present researches, to use scales that can measure the impact of swallowing training in the nutritional and pulmonary condition of dysphagic patients. An important research area, related to the control of therapeutic efficacy and efficiency, are the studies that aim to establish the decrease in hospital and home care costs as a consequence of speech language intervention with patients with oropharyngeal dyspahgia. PMID- 17461356 TI - [Lung sonography for the cardiologist]. AB - In the recent years lung sonography has overcome his pioneer phase with the contribution of significant data from authors of different disciplines. Today there is therefore a strong evidence that this diagnostic method could have an important impact for the diagnosis of cardiothoracic symptoms, mainly dyspnea, in emergency or in ambulatory medicine. Since the cardiologist already uses ultrasound for the study of the heart, he or she could consider the integrated cardiopulmonary sonography extremely attractive in this specific context. In this review the authors describe the anatomical, biophysical and clinical basis of lung ultrasonography in a cardiological setting. PMID- 17461357 TI - [Risk factors for cardiovascular diseases: what is the role for homocysteine?]. AB - Cardiovascular diseases are commonly related to classical risk factors, but other risk markers have been identified, including homocysteine. Homocysteine is a sulphurated amino acid which derives from methionine. The causes of hyperhomocysteinemia are multifactorial, such as genetic defects, pathophysiological conditions, lifestyle and drugs-related. Hyperhomocysteinemia favors atherothrombosis through endothelial dysfunction, enhancement of inflammation and thrombophilic profile. A number of clinical and laboratory trials exist regarding the association between homocysteine levels and an increased risk of cardiovascular disease. However, the lack of homogeneity in the data, together with the high number of factors capable of influencing homocysteine plasma levels, and the disappointing results of therapeutic trials do not permit us at present to consider homocysteine as an independent and major risk factor for cardiovascular disease. PMID- 17461358 TI - [Electrocardiographic diagnosis of left ventricular hypertrophy in the presence of intraventricular conduction disturbances]. AB - It is commonly agreed that the electrocardiographic recognition of left ventricular hypertrophy (LVH) is difficult, or even impossible, in patients with bundle branch or fascicular block; the opposite, however, has been demonstrated by several studies. In the presence of intraventricular conduction disturbances, many criteria can reveal LVH, with sensitivity and specificity not inferior than that of electrocardiographic signs used in subjects with normal intraventricular conduction. The following criteria can be helpful in left bundle branch block: QRS voltage increase, left atrial enlargement, QRS duration > 155 ms. LVH is suggested by one or more of the following: Sokolow index > or = 35 mm, R wave in lead aVL > or = 11 mm, left axis deviation at -40 degrees or more, SV2 > 30 mm + SV3 >25 mm. In left anterior hemiblock, LVH is diagnosed whenever the sum of S wave in lead III plus the maximal R+S in a precordial lead is > or = 30 mm. Further criteria are SV1 + (R+S) in V5 or V6 > or = 25 mm, and the presence of secondary ST-T changes. In right bundle branch block, LVH is suggested by a left atrial enlargement pattern, secondary repolarization changes, and a sum of S wave in lead III plus the maximal R+S in a precordial lead > or = 35 mm. PMID- 17461359 TI - [Teleconsulting in echocardiography using a standard cable on HDSL]. AB - BACKGROUND: Teleconsultation in echocardiography is complex owing to the heavy images and video files to be exchanged; moreover standards for data acquisition still need to be defined. The aim of this experience was to verify the feasibility to connect two laboratories for teleconsulting using a standard HDSL telephone cable. METHODS: Hardware included one echocardiograph, one PC, video aquisition board and network board, USB webcam, Network router and an HDSL line for data communication. Software includes MS Windows XP and Netmeeting. The live interactivity between the labs was granted by a video streaming in MPEG4 format, directly delivered by the PC communication software. Standard DICOM was used for formal aspects. The platform has been verified during a demo at "National Congress of Cardiovascular Echography" in 2005. RESULTS: The experiment survey collected at the Teleconference gave a consistently good result. Twenty-one subjects filled in the questionnaire: 12 of them (57%) reported a difference between the video streaming and DICOM; whereas 7 of them (33%) did not point out it. Overall findings have been positive, even though 15 members (71%) reported feasibility with some difficulties; 18 members (86%) reported the possibility of really using this technology in the future. The total cost was 145000 euro. CONCLUSIONS: In our opinion, overall cost is more than acceptable especially considering how often to send data is more convenient than moving sick people or consultants. PMID- 17461360 TI - [Car driving after cardiac surgery: clinical aspects, regulations, and legal implications]. AB - BACKGROUND: Car driving is one of the most perceived problems by patients after cardiac surgery. The aim of this study was to evaluate the influence of cardiac surgery and median sternotomy on driving performance after a cardiac rehabilitation program. METHODS: Seventy-four consecutive patients, usual car drivers, admitted to our Cardiac Rehabilitation Center after cardiac surgery, were evaluated 60 days from discharge using a five-item questionnaire. Questions were related to resume of car driving, problems related to car driving, and seatbelt wearing. RESULTS: The analysis of the answers indicated that 36% of patients (62% females, 26% males; p = 0.01) stopped car driving after cardiac surgery, owing to the lack of security or fear of either the patients or their relatives. Sixty-four patients continued to drive, however 39% of them reported problems related to driving (irritation, poor concentration, fear, parking maneuvers). Because of persistent post-surgical chest pain, 15% of patients avoided wearing seatbelts, and 7% asked for exclusion following unconventional procedures. CONCLUSIONS: After cardiac surgery, 1 out of 3 patients stops car driving, particularly among female gender; 1 out of 7 patients is seatbelt no wearers and about half of them claim for exemption following procedures not codified. Thus, common rules and recommendations from Scientific Societies are highly warranted. PMID- 17461361 TI - [Italian clinical trials on cardiac cell therapy: where we are and where are we going?]. AB - BACKGROUND: In the last years translation from bench to bedside of findings regarding cardiac cell therapy is swinging between delays and accelerations. Based on experimental studies, clinical trials were started in 2001. To date in Europe more than 900 patients have been treated with cell transplantation or mobilization and new clinical trials are ongoing in many countries. Published data provide a limited idea of current Italian clinical research in this field. This investigation intends to report Italian clinical trials and projects using cell therapy in cardiology. METHODS: Italian Centers involved in cell therapy clinical studies have been detected exploring scientific bibliography and online search engine. Selected Centers have been classified by the origin of collected information: i) peer-reviewed journals; ii) abstract, poster or communications; iii) press or media. A questionnaire was used and face-to-face or telephonic interviews were performed to obtain further details where needed. RESULTS: Among the 16 selected Centers, 5 published their experiences on peer-reviewed journals, 5 produced abstracts or communications for scientific meetings and 6 bring out their protocols through press or media. Usually Italian clinical trials are monocenter (11 monocenter studies, 2 multicenter) and enrol few patients (an average of about 10 patients treated in each study); 2 Centers are waiting for protocol approval by the Italian Institute of Health; in one case approval was not obtained by the Ethics Committee. Few centers found projects on their own preclinical studies. CONCLUSIONS: The investigation on Italian clinical experiences with cardiac cell therapy demonstrates the limits of what has been produced so far. An Italian working group on cardiovascular regenerative therapies could represent a useful tool to improve national clinical research in this field. PMID- 17461362 TI - [A case of right ventricular ischemia mimicking acute myocardial infarction during coronary angioplasty on the right coronary artery]. AB - A 55-year-old man was admitted to our department with a diagnosis of acute inferior myocardial infarction; 1 week later he underwent percutaneous transluminal coronary angioplasty on the proximal right coronary artery; during this procedure, the patient experienced chest pain and his electrocardiogram showed ST-segment elevation in the anterior leads; ventricular fibrillation also occurred. The ST-segment abnormality disappeared within 45 min, and no stenosis or spasm on the left coronary artery were detected during angiographic control; the blood flow in the right coronary artery was normal, but the acute marginal branch was occluded. This is a rare case of right ventricular ischemia caused by occlusion of the acute marginal branch during coronary angioplasty on the right coronary artery. PMID- 17461363 TI - [Primary cardiac sarcoma. Description of a case]. AB - Primary cardiac tumors are rare events. We describe here a case of undifferentiated pleomorphic sarcoma (so-called pleomorphic malignant fibrous histiocytoma) obliterating mostly the left side and the anterior wall of pericardium in a 84-year-old man admitted for mild dyspnea at rest. The diagnosis was suspected after excluding the lung origin of the mass (observed by plain chest radiography) by thorax computed tomography but it was confirmed only by cardiac-gated magnetic resonance imaging and transthoracic biopsy. Considering both patient's age and comorbidity, and local extension of the lesion, after counseling with cardiac surgeons and oncologists, the patient was treated only by conservative medical therapy. The patient died 6 months after the diagnosis due to a superior vena cava syndrome as an effect of infiltration and obstruction of superior vena cava by the tumor at the site of vein entry in the right atrium. This case is an example of a primary cardiac tumor that causes relative myocardial sufferance both by infiltration and by limitation of normal heart diastolic function. PMID- 17461364 TI - [Large saddle pulmonary artery thrombus visualized by transthoracic echocardiography]. PMID- 17461365 TI - [Coronary surgery on the beating heart]. PMID- 17461366 TI - Can psychiatric intervention improve major depression in very near end-of-life cancer patients? AB - OBJECTIVE: Although depression is a prevalent and burdensome psychiatric problem in end-of-life cancer patients, little is known about its susceptibility to treatment, especially when patients reach very close to the end of life. This study was conducted to evaluate response rate of that end-of-life depression to psychiatric intervention and to assess the feasibility of conventional evidence based pharmacological therapy for depression. METHODS: The medical records of 20 patients who were referred to the psychiatry division for major depressive disorder and died within 3 months after the referral were reviewed. The Clinical Global Impression-Improvement (CGI-I) Scale was used for each case, and responders were defined as patients whose scores were much or very much improved. All pharmacological treatments were extracted, and the doses of the antidepressant prescribed were compared to their evidence-based-defined therapeutic doses. RESULTS: Of the 20 patients, seven were responders, but no response was achieved when the survival time was less than 3 weeks. Most patients were treated with antidepressants, but the doses prescribed were far less than the defined doses, especially the doses of the tricyclic antidepreSsants (TCAs). SIGNIFICANCE OF RESULTS: These results suggested that patients' survival time largely determines susceptibility to psychiatric treatment, and it is hard to achieve response in patients whose survival time was less than about 1 month. Implementation of conventional evidence-based pharmacological treatment is difficult, especially with TCAs, and various antidepressants, which can be administrated by other routes, are needed when oral intake is impossible. PMID- 17461367 TI - Caregiving at the end of life: perspectives from spousal caregivers and care recipients. AB - OBJECTIVE: The purpose of this study is to examine the perspectives of both the spousal caregiver and care recipient on the caregiving experience in home-based palliative care. METHODS: A qualitative research strategy involving home-based face-to-face interviews with older palliative care patients and their spousal caregivers was used to examine the caregiving experience. RESULTS: Ten spousal caregivers and care recipient dyads participated in the study. Most informal caregivers viewed caregiving as an extension of the family relationship where caregiving responsibilities evolved over time. Spousal caregivers identified many negative reactions to caregiving, such as fatigue or weariness, depression, anger and sadness, financial stresses, and lack of time. Care recipients acknowledged the emotional and financial strain and expressed concern for their spouses. Both caregivers and care recipients were appreciative of home care services although they identified the need for additional services. They also identified difficulties in communication with formal providers and poor coordination of care among the various services. Both caregivers and care recipients disclosed some challenges with informal supports, but on the whole felt that their presence was positive. Additional positive aspects of caregiving reported by spouses included strengthened relationship with their spouse and discovering emotional strength and physical abilities in managing care. SIGNIFICANCE OF RESULTS: Health care and social service professionals need to recognize and understand both caregiver and care recipient perspectives if they are to successfully meet the needs of both members of the dyad. PMID- 17461368 TI - Factors relating to terminally ill cancer patients' willingness to continue living at home during the early phase of home care after discharge from clinical cancer centers in Japan. AB - OBJECTIVE: To assess the willingness of Japanese terminally ill cancer patients to continue living at home during the early phase of home care after discharge from a Clinical Cancer Center (CCC) in Japan, and to identify factors relating to their willingness to continue living at home. METHODS: A cross-sectional questionnaire survey of a convenient sample of both Japanese terminally ill cancer patients and their caregivers (PFCs) was conducted (n = 294, effective response rate 25.0%). Questionnaires were mailed and medical records were accessed for 73 pairs of respondents, comprising one terminally ill cancer patient and one PFC. RESULTS: At about 10 days after discharge, 64 patients (88%) wished to continue living at home. A hierarchical logistic regression analysis was performed on the data. It was found that the fewer the medical treatments undergone (OR = 0.20, 95% CI: 0.05-0.72), the higher the patients' perception that their condition was consistent with care at home (OR = 2.77, 95% CI: 1.08 8.62) and with their functional well-being (OR = 1.45, 95% CI: 1.08-2.17). In addition, the higher the caregivers' satisfaction with life (OR = 2.37, 95% CI: 1.15-5.77), the more willing patients tended to be to continue living at home. SIGNIFICANT OF RESULTS: The willingness of Japanese terminally ill cancer patients to continue living at home appears to be affected by caregiver status. This indicates a need for discharging facilities to monitor the state of home assistance and to investigate the nature of assistance required for continuing home care. PMID- 17461369 TI - Struggling in change at the end of life: a nursing inquiry. AB - OBJECTIVE: The purpose of this human science nursing inquiry is to explore the meaning of struggling in change for persons at the end of life. METHODS: Guided by Parse's theory of human becoming, a descriptive exploratory method was used to answer the research question: What is the meaning of the experience of struggling in change for persons at the end of life? Eight persons who were living with dying described experiences of struggling in change during face-to-face audiotaped interviews. RESULTS: A process of analysis-synthesis revealed three themes that are discussed in relation to extant related literature and interpreted in light of the human becoming perspective. SIGNIFICANCE OF RESULTS: Findings from the study contribute new knowledge about human experience at the end of life from a human science perspective and offer new insights on struggling in change as a rhythmical pattern of living and dying. Implications for palliative practice, research, and education are discussed. PMID- 17461370 TI - From "invincibility" to "normalcy": coping strategies of young adults during the cancer journey. AB - OBJECTIVE: Little research has been undertaken regarding the psychological impact of cancer on those stricken during the young adult years. Specifically, research on the coping strategies of young adults with cancer is limited. METHOD: In this qualitative, Grounded Theory study, we did not set out to examine coping; rather, it emerged as a major phenomenon in the analysis of interview data from 15 young adults with cancer. RESULTS: These young adults used various coping strategies to come to terms with the cancer diagnosis, management of the illness, its treatment, and treatment sequelae. The coping strategies varied considerably from person to person, depended on the stage(s) of the illness, and were rooted in their precancer lives. We were able to discern a pattern of coping strategies used by most participants. The prevailing goal for all participants was to achieve what they called "normalcy." For some, this meant major changes in their lives; for others it meant to "pick up" where they had left off before the cancer diagnosis. SIGNIFICANCE OF RESULTS: To aid the understanding of the issues that influence coping, we have developed a model to illustrate the bidirectional nature and the complexities of the coping strategies as they relate to the phases of the disease and the disease treatment. The model also affirms Folkman and Lazarus' coping theory. PMID- 17461371 TI - Crying: a force to balance emotions among cancer patients in palliative home care. AB - OBJECTIVE: Crying is a common but seldom studied phenomenon in palliative care. The aim of this study was to explore the significance of patients crying in a palliative care context. METHODS: Tape-recorded interviews with 14 cancer patients in palliative home care were carried out. To gain deeper understanding, a hermeneutic analysis and interpretation was used. RESULTS: Crying was described in different dimensions: (1) intense and despondent crying as a way of ventilating urgent needs, (2) gentle, sorrowful crying as a conscious release of emotions, and (3) quiet, tearless crying as a protection strategy. Crying seems to be an expression for an inner emotional force, provoked by different factors, which cause changes in the present balance. To cry openly but also to cry on the inside meant being able to achieve or maintain balance. Crying may be something useful, which could create release and help reduce tension, but it may also have a negative impact as it consumes energy and creates feelings of shame. SIGNIFICANCE OF RESULTS: Professionals need to understand the different levels of crying. In such situations sometimes comforting the patient may not be the best solution, as some may need privacy. PMID- 17461372 TI - Does social support from family and friends work as a buffer against reactions to stressful life events such as terminal cancer? AB - OBJECTIVE: To examine the relationship between social support and emotional functioning and stress reactions. Our hypothesis is that patients who reported a high degree of social support will experience better emotional functioning and less serious stress reactions than patients with a low degree of social support. METHOD: The sample was comprised of 434 patients at the Palliative Medicine Unit (PMU), University Hospital of Trondheim in Norway. The patients completed a questionnaire monthly including questions about social support from the MacAdam's Scale, subjective stress measured by the Impact of Event Scale (IES), and emotional functioning measured by the subscale in the EORTC QLQ-30. RESULTS: Although our hypothesis was not supported at the baseline assessment, it was supported at the second assessment, 2 months later. Patients with high social support reported better emotional functioning and less serious stress reactions, in terms of lower scores on the IES avoidance subscale, than patients with a low degree of social support. SIGNIFICANCE OF THE RESULTS: The mixed findings may indicate that social support has only small effects on emotional functioning and stress reactions. Our results on the second assessment indicate, however, that social support might work as a buffer against reactions toward external stressful events such as terminal cancer. PMID- 17461373 TI - The physician's disease: the impact of medical knowledge on personal illness. AB - OBJECTIVE: When physicians confront a personal illness of a serious nature, they may discover that the transition to the sick role is challenging, and the inability to relinquish their stethoscope may cause undo anxiety. The physician patient relationship is intrinsically asymmetrical, and the role of the physician is to regulate the amount of information patients need in order to become educated about their illness and to make informed decisions about their treatment plan. This article explores the challenges in the physician-patient relationship when the patient is also a physician. METHODS: This article is a literature review of publications involving the unique challenges physician-patients experience when suffering from serious personal illness. RESULTS: The medical knowledge physician-patients harbor has the potential to complicate their ability to cope with difficult or terminal diagnoses. Paradoxically, knowledge about a condition may fuel anxiety instead of alleviating the fear associated with the unknown. Medical knowledge therefore may entail a certain loss of innocence, and physicians are often unable to revert to being "mere" patients. Furthermore, managing this anxiety in physician-patients may prove to be challenging to the treating physician. SIGNIFICANCE OF RESULTS: From a medical perspective, physician-patients need to be addressed like any other patient. Psychologically, however, these patients are unique, and the specific challenges their education and experience bring into the consultation room needs to be explicitly addressed. PMID- 17461374 TI - Crib death. Dark side of the world's greatest job. PMID- 17461375 TI - Toxic or hypoxic? Is the patient one, the other or both? PMID- 17461376 TI - Medicare appeals process less than appealing. PMID- 17461377 TI - Where does EMS fit in the federal landscape? PMID- 17461378 TI - Fracture management. Effective prehospital splinting techniques. AB - Fractures are commonly encountered in the prehospital setting. Using some very basic methods and equipment, as well as some advanced equipment options, prehospital providers have the ability to recognize and treat many of these fractures. PMID- 17461379 TI - A milestone change in practice: a call for widespread application of intranasal medication delivery in the prehospital environment. PMID- 17461380 TI - Assessment and treatment of ankle injuries. PMID- 17461381 TI - Achieving legislative success: a summit approach to collaboration. PMID- 17461382 TI - Beyond the basics: trends and changes in pediatric care. PMID- 17461383 TI - You take my breath away. PMID- 17461384 TI - Transporting children. PMID- 17461385 TI - Pediatric care and EMS. An interview with Tommy Loyacono. Interview by Raphael M Barishansky. PMID- 17461386 TI - Coping with violent people: a multi-part series. PMID- 17461387 TI - A guide to hazmat response. AB - The Emergency Response Guidebook provides responders with important first response information. It is intended to assist in managing the first 15-30 minutes of an emergency response and should not be used as the definitive response resource. It does, however, provide information that can be used to identify protective clothing and response procedures that can save lives and initiate successful control of the incident. Using the known product information, responders can use the ERG to determine response procedures for emergencies involving chemicals in a fixed facility. Responders should review and practice using the guidebook on a regular basis. PMID- 17461388 TI - Scene safety for bike medics. PMID- 17461389 TI - Leadership Tips. Cards: thanking people's families. PMID- 17461390 TI - [Prevalence of hereditary diseases in three-year-old horses of the Freiberger breed]. AB - The objective of this study was to investigate clinical signs indicating diseases with known or suspected hereditary components like equine sarcoid, insect bite hypersensitivity, osteochondrosis, allergic airway diseases, podotrochleosis, prognatism and wind-sucking in the franches-montagnes horse. We performed a clinical examination on 702 three-year-old, privately owned franches-montagnes horses, which were shown at the Swiss-Field-Tests in 2004. With the help of the owners a questionnaire on health, environment and feeding habits of the animals was completed. In 11.9% of the horses, sarcoids were detected, 6.2% carried one tumor and 5.7% had several. The prevalence was higher in chestnuts (16.6%) than in bays (10.1%). The prevalence of sarcoids in offspring from sires with known sarcoids (data from Station-Tests 1994-2005) was not significantly higher than in descendants from stallions without a known history of sarcoids. Clinical signs of insect bite hypersensitivity were only found in six horses (0.9%). In 12.0% hoof abnormalities like brittle horn, ring hoofs or hoof cracks of different degrees were recorded. The angle between hoof base and hoof wall was 56.7 +/- 0.1 degrees, the average hoof width was 13.7 +/- 0.3 cm in the front feet. For both of these measures we found no significant difference between left and right feet. With the exception of a high sarcoid prevalence, our results indicate that the franches-montagnes horse is a healthy breed overall. PMID- 17461391 TI - [Prevalence of hereditary diseases in three-year-old Swiss Warmblood horses]. AB - The objective of this study was to investigate clinical signs indicating hereditary diseases like equine sarcoid, osteochondrosis (OC) and the idiopathic laryngeal hemiplegia (ILH), and to demonstrate relationships between environment, feeding habits and conformation ("exterieur" evaluation) of the horses. For this purpose, we analyzed veterinary examinations of 403 stallions at the approvals since 1994 examined 493 three-year-old Swiss Warmblood horses, which were shown at the Swiss-Field-Tests in 2005. With the help of the owners a questionnaire on health, environment and feeding habits of the animals was completed. At the same time, the horses were assessed and graded for their "exterieur" (type, conformation, gaits) by judges of the Swiss Sporthorse breeding association. In 11.5% of horses sarcoids were found, 8.7% showed one and 2.8% several tumors. The prevalence of sarcoids in offspring of sires with known sarcoids was not significantly higher than in descendants from stallions without a known history of sarcoids. We found distended joints as a possible symptom of OC in 11.4% of the horses, 3.9% (n = 19) in both tarsal joints. We did not find a relationship between enlarged joints in the offspring and the presence of OC in the sires. Abnormal respiratory noise at work, as a possible sign for ILH, was heard only in 1.2% (n = 6). It is important to note that while we found a high number of sarcoid affected horses compared to other studies, presence of enlarged joints was not very frequent and very few horses showed abnormal respiratory noise. Additionally, we found no correlation between "exterieur" marks and the horse's general health. PMID- 17461393 TI - Modified skew-flap below-knee amputation. AB - Between 1999 and 2001, 35 consecutive patients with diabetes (mean age, 59.4 years) were treated prospectively with a modified skew-flap below-knee amputation. The technique, results, and follow-up are described. By a mean follow up of 3.5 years, 3 patients required below-knee amputation of the opposite extremity, 4 expired, and 28 were ambulating with a below-knee prosthesis. The modification has several advantages: A tibialis anterior muscle cushion on the distal end of the tibia prevents bone protrusion; anterior skin flaps made by the initial linear anterior incision prevent tension at the suture line; and oblique myocutaneous flaps avoid muscle trimming and prevent shearing of fascial plexuses at closure, thus improving wound healing. PMID- 17461394 TI - Evaluating short-term pain after steroid injection. AB - Steroids are injected into joints for various indications. All steroid preparations relieve pain similarly over the long term. Therefore, decisions about which preparation to use are often arbitrary. We evaluated methylprednisolone acetate and a combination of betamethasone diproprionate and betamethasone sodium phosphate for short-term pain and the predictive value of short-term pain. Eighty-five patients were injected in prospective double-blind randomized fashion. Pain was evaluated by visual analog scale (1 = no pain, 10 = severe pain) at baseline, 3 days, and 3 weeks. No patient had joint pain immediately after injection. Three days after injection, mean (SD) pain levels were 5.1 (2.9) for methylprednisolone and 5.2 (2.6) for betamethasone (P = .97); 3 weeks after injection, they were 4.0 (2.8) and 3.7 (2.5), respectively (P = .57). Short-term pain increased from baseline for both preparations and decreased from 3 days to 3 weeks. Pain at 3 days and 3 weeks was positively correlated. This study does not support a difference in short-term pain between preparations. The significant correlation between short- and long-term pain may justify early decisions regarding treatment, especially in patients with high levels of initial pain. PMID- 17461396 TI - Anatomic variations in the lateral femoral cutaneous nerve with respect to pediatric hip surgery. AB - Variations were documented in the course of the lateral femoral cutaneous nerve (LFCN) in the upper thigh relative to anatomic landmarks in 22 adult cadavers using the Smith-Petersen incision for the anterior approach to the hip. Distances from the anterior superior iliac spine (ASIS) to the point of nerve entry into the thigh were normalized as percentages of the distance from the ASIS to the pubic tubercle (PT) to relate the data to small children. In all cases, the LFCN passed deep to the inguinal ligament, entering the thigh a mean of 2.6 cm (SD, 1.9 cm) medial from the ASIS (19%+/-14% of the ASIS-PT distance), with distances ranging from 0.3 to 6.5 cm (2.6%-46.4%). With the data extrapolated to children, the LFCN may commonly be found medial to the ASIS about one fifth the distance from the ASIS to the PT. In 32% of cases, the LFCN ran directly inferiorly, but in 68% it coursed inferolaterally and then turned to run inferiorly close to the distal part of the incision. Expressed proportionally rather than only as mean measurements, these percentages provide a better estimate of the location of the LFCN in relation to patient size and thus are useful when operating in this region. PMID- 17461395 TI - Thromboprophylaxis after hip fracture: evaluation of 3 pharmacologic agents. AB - We compared the clinical efficacy and side-effect profiles of aspirin, dextran 40, and low-molecular-weight heparin (enoxaparin) in preventing thromboembolic phenomena after hip fracture surgery. All patients admitted with a diagnosis of hip fracture to our institution between July 1, 1987, and December 31, 1999, were evaluated. Study inclusion criteria were age 65 years or older, previously ambulatory, cognitively intact, home-dwelling, and having a nonpathologic intertrochanteric or femoral neck fracture. Each patient received mechanical thromboprophylaxis (above-knee elastic stockings) and 1 pharmacologic agent (aspirin, dextran 40, or enoxaparin); patients who received aspirin were also given a calf sequential compression device. Meeting the selection criteria and included in the study were 917 patients. Findings included low incidence of thromboembolic phenomena (deep vein thrombosis, 0.5%-1.7%; pulmonary embolism, 0% 2.0%; fatal pulmonary embolism, 0%-0.5%) and no difference among the 3 pharmacologic agents in thromboembolic prophylaxis efficacy. Use of enoxaparin was associated with a significant increase (3.8%) in wound hematoma compared with dextran 40 (1.6%) and aspirin (2.4%) (P<.01). The 3 agents were found not to differ with respect to mortality, thromboembolic phenomena, hemorrhagic complications, or wound complications. PMID- 17461397 TI - Subtalar dislocation in an 8-year-old boy: a rare clinical presentation. PMID- 17461398 TI - Incidence of os acromiale in patients with shoulder pain. AB - A prospective case series was undertaken to determine the incidence of os acromiale in patients presenting to an orthopedic clinic with shoulder pain during a 6-month period. Ninety-three shoulders in 88 consecutive patients were evaluated by history, physical examination, and radiographs. Radiographs included anteroposterior, scapular Y, and axillary views in all patients. Based on history and physical examination, no patient was given a primary diagnosis of os acromiale. However, when the plain films were specifically reviewed for this entity, it was present in 6 (6.82%) of the 88 patients. It was seen on the axillary view in each case. Five (83.3%) of the 6 patients had been given a diagnosis of either impingement or rotator cuff tear based on physical examination. Os acromiale is an infrequent but not rare entity that must be kept in mind when evaluating patients with shoulder pain, as failure to recognize an os acromiale may negatively influence treatment results. An axillary view should be obtained when evaluating patients with shoulder pain. PMID- 17461399 TI - Outsourcing your billing. PMID- 17461400 TI - Combined technique for draining septic arthritis of the pediatric hip. AB - Recent literature suggests that, in older children, acute cases of septic arthritis may be treated with aspiration and intravenous antibiotics. However, when surgical decompression is required, the technique described in this report has several advantages. It can be performed with an incision of approximately 1 to 2 inches and a posterior stab wound of <1 cm; it preserves anatomic planes and poses less risk to the circumflex femoral vessels (as with the anterior approach); and it allows fluid to drain when the patient is supine (as with the posterior approach). PMID- 17461401 TI - Raise your workplace's activity level. PMID- 17461402 TI - Time to be clutter-free. PMID- 17461403 TI - Man overboard! PMID- 17461404 TI - Mobilizing safety leadership and performance. PMID- 17461405 TI - Walk the flight line. PMID- 17461406 TI - A rising tide. PMID- 17461407 TI - Training the industrial athlete: fitness training at UPS. PMID- 17461408 TI - Working hand in hand. PMID- 17461410 TI - The first cut is the deepest. PMID- 17461411 TI - Frequently asked questions: ergonomics and hand protection. PMID- 17461413 TI - Key factors in fitting. PMID- 17461412 TI - Why safety training may not be the answer. PMID- 17461414 TI - MRSR Staph infections: coming to a workplace near you. PMID- 17461415 TI - Complying with OSHA's fire safety standards. PMID- 17461416 TI - Regulation of Toll-like receptor 4 expression and its signaling by hypoxia in cultured microglia. AB - Hypoxia is an important biological signal that regulates a wide variety of physiological responses. At the same time, hypoxia is involved in multiple pathological situations. In particular, hypoxia is closely associated with neural injury in the brain. Hypoxia has been recently proposed as a neuroinflammatogen, as it can induce the inflammatory activation of microglia, a major cellular source of inflammatory mediators in the brain. In this article, we present evidence that hypoxia enhances Toll-like receptor 4 (TLR4) expression in cultured microglia and differentially regulates the downstream signaling pathways of TLR4. Hypoxia up-regulated TLR4 expression at the mRNA and protein levels in a microglia cell line, as well as in primary microglia cultures. Hypoxia, however, differentially regulated MyD88-dependent and -independent pathways of TLR4 signaling: Hypoxia enhanced lipopolysaccharide (LPS)-induced interferon regulatory factor-3 (IRF-3) activation and the subsequent expression of IFNbeta (MyD88-independent pathway), whereas it suppressed LPS-induced NF-kappaB activation (MyD88-dependent pathway). Hypoxia did not affect IFNgamma signaling, which was represented by signal transducer and activator of transcription-1 (STAT1) activation and interferon-regulatory factor-1 (IRF-1) induction. Taken together, although hypoxia up-regulates TLR4 expression, its downstream signaling pathways appear to be differentially modulated by hypoxia. PMID- 17461417 TI - Acutely increased cyclophilin a expression after brain injury: a role in blood brain barrier function and tissue preservation. AB - Blood-brain barrier (BBB) compromise is a significant pathologic event that manifests early following traumatic brain injury (TBI). Because many signaling cascades are initiated immediately after the traumatic event, we were interested in examining acute differential protein expression that may be involved in BBB function. At acute time points postinjury, altered protein expression may result from altered translation efficiency or turnover rate rather than from a genomic response. The application of tandem 2-D gel electrophoresis and mass spectrometry analysis is a powerful approach for directly screening differential protein expression following TBI. Using comparative 2-D gel analysis, we selected candidate protein spots with apparent altered expression and identified them by mass spectrometry. Cyclophilin A was selected for further analysis because it has been implicated in endothelial cell activation and inflammation, and studies have suggested cyclosporine A, an inhibitor of all cyclophilin isoforms, might be beneficial after TBI. We examined if altered expression of cyclophilin A in the brain vasculature might play a role in BBB function. We found significantly increased cyclophilin A levels in isolated brain microvessels 30 min following injury. Postinjury administration of cyclosporine A significantly attenuated BBB permeability measured 24 hr postinjury, suggesting cyclophilin activity after TBI may be detrimental. However, direct injection of purified recombinant cyclophilin A attenuated both BBB permeability and tissue damage in a stab wound model of injury. These findings suggest that increased expression of cyclophilin A may play a protective role after TBI, whereas other cyclophilin isoforms may be detrimental. PMID- 17461418 TI - A 7 gene signature identifies the risk of developing cirrhosis in patients with chronic hepatitis C. AB - Clinical factors such as age, gender, alcohol use, and age-at-infection influence the progression to cirrhosis but cannot accurately predict the risk of developing cirrhosis in patients with chronic hepatitis C (CHC). The aim of this study was to develop a predictive signature for cirrhosis in Caucasian patients. All patients had well-characterized liver histology and clinical factors; DNA was extracted from whole blood for genotyping. We validated all significant markers from a genome scan in the training cohort, and selected 361 markers for the signature building. Using a "machine learning" approach, a signature consisting of markers most predictive for cirrhosis risk in Caucasian patients was developed in the training set (N = 420). The Cirrhosis Risk Score (CRS) was calculated to estimate the risk of developing cirrhosis for each patient. The CRS performance was then tested in an independently enrolled validation cohort of 154 Caucasian patients. A CRS signature consisting of 7 markers was developed for Caucasian patients. The area-under-the-ROC curves (AUC) of the CRS was 0.75 in the training cohort. In the validation cohort, AUC was only 0.53 for clinical factors, increased to 0.73 for CRS, and 0.76 when CRS and clinical factors were combined. A low CRS cutoff of <0.50 to identify low-risk patients would misclassify only 10.3% of high-risk patients, while a high cutoff of >0.70 to identify high-risk patients would misclassify 22.3% of low-risk patients. CONCLUSION: CRS is a better predictor than clinical factors in differentiating high-risk versus low risk for cirrhosis in Caucasian CHC patients. Prospective studies should be conducted to further validate these findings. PMID- 17461419 TI - InXy and SeXy, compact heterologous reporter proteins for mammalian cells. AB - Mammalian reporter proteins are essential for gene-function analysis, drugscreening initiatives and as model product proteins for biopharmaceutical manufacturing. Bacillus subtilis can maintain its metabolism by secreting Xylanase A (XynA), which converts xylan into shorter xylose oligosaccharides. XynA is a family 11 xylanase monospecific for D-xylose containing substrates. Mammalian cells transgenic for constitutive expression of wild-type xynA showed substantial secretion of this prokaryotic enzyme. Deletion analysis confirmed that a prokaryotic signal sequence encoded within the first 81 nucleotides was compatible with the secretory pathway of mammalian cells. Codon optimization combined with elimination of the prokaryotic signal sequence resulted in an exclusively intracellular mammalian Xylanase A variant (InXy) while replacement by an immunoglobulin-derived secretion signal created an optimal secreted Xylanase A derivative (SeXy). A variety of chromogenic and fluorescence-based assays adapted for use with mammalian cells detected InXy and SeXy with high sensitivity and showed that both reporter proteins resisted repeated freeze/thaw cycles, remained active over wide temperature and pH ranges, were extremely stable in human serum stored at room temperature and could independently be quantified in samples also containing other prominent reporter proteins such as the human placental alkaline phosphatase (SEAP) and the Bacillus stearothermophilus-derived secreted alpha-amylase (SAMY). Glycoprofiling revealed that SeXy produced in mammalian cells was N- glycosylated at four different sites, mutation of which resulted in impaired secretion. SeXy was successfully expressed in a variety of mammalian cell lines and primary cells following transient transfection and transduction with adeno-associated virus particles (AAV) engineered for constitutive SeXy expression. Intramuscular injection of transgenic AAVs into mice showed significant SeXy levels in the bloodstream. InXy and SeXy are highly sensitive, compact and robust reporter proteins, fully compatible with pre-existing marker genes and can be assayed in high-throughput formats using very small sample volumes. PMID- 17461420 TI - A genetic time-delay circuitry in mammalian cells. AB - Gene expression circuitries with time-delayed expression profiles regulate key events, such as oscillating systems, noise elimination, and coordinated multi step processes, in all organisms from bacteria to mammalian cells. We present the rational synthesis of a genetic circuit displaying time-delayed expression in silico and in mammalian cells. The network is based on a time-delay circuit, where the tetracycline-responsive transactivator (tTA) induces expression of the pristinamycin-responsive repressor PIP-KRAB, which silences expression of the terminal human placental secreted alkaline phosphatase (SEAP). While the addition of pristinamycin I inactivates PIP-KRAB and results in the immediate resumption of SEAP expression, addition of tetracycline abolishes PIP-KRAB synthesis. Consequently, SEAP production remains repressed until the PIP-KRAB buffer in the cell is eliminated. We characterized in silico and in vivo the time-delayed expression properties and analyzed the impact of the size and stability of the PIP-KRAB buffer on fine-tuning of the response kinetics. This tunable time-delay circuitry represents a biologic building block for emulating a fundamental circuit topology in integrated artificial synthetic gene networks for the design of tailor-made cell types and organisms. PMID- 17461421 TI - Influence of microbial growth kinetics on steady state multiplicity and stability of a two-step nitrification (SHARON) model. AB - In this paper, the influence of microbial growth kinetics on the number and the stability of steady states for a nitrogen removal process is addressed. A two step nitrification model is studied, in which the maximum growth rate of ammonium oxidizers is larger than the one of nitrite oxidizers. This model describes the behavior of a SHARON reactor for the treatment of wastewater streams with high ammonium concentrations. Steady states are identified through direct calculation using a canonical state space model representation, for several types of microbial kinetics. The stability of the steady states is assessed and the corresponding phase portraits are analyzed. Practical operation of a SHARON reactor aims at reaching ammonium conversion to nitrite while suppressing further conversion to nitrate. Regions in the input space are identified that result in this desired behavior, with only nitrite formation. It is demonstrated that not only the dilution rate plays a role, as is commonly known, but also the influent ammonium concentration. Besides, the type of microbial (inhibition) kinetics has a nonnegligible influence. While the results indicate that product inhibition does not affect the number of steady states of a (bio)reactor model, it is shown that substrate inhibition clearly yields additional steady states. Particular attention is devoted to the physical interpretation of these phenomena. PMID- 17461422 TI - Effect of moderate electric field on the metabolic activity and growth kinetics of Lactobacillus acidophilus. AB - Moderate electric fields (MEF), applied across microbial growth media may potentially affect the permeability of cell membranes. We investigated the effects of MEF on bacteriocin (lacidin A) production during fermentation and on microbial growth kinetics of Lactobacillus acidophilus OSU 133. We comparatively investigated the following treatments: conventional, MEF (1 V/cm, 60 Hz, for 40 h), combinations of MEF (1 V/cm, 60 Hz, for the first 5 h) and conventional fermentation (for 35 h), and discrete MEF (1 V/cm, 2 min on and off, for 40 h). In all treatments, except as noted below, temperature was set at 30 degrees C. The two exceptions were control (conventional) and discrete MEF treatment, which were conducted both at 30 and 37 degrees C. MEF treatments at the early stage of fermentation at 30 degrees C showed the maximum bacteriocin activity. Minimum bacteriocin production was observed under conventional fermentation at 37 degrees C. A mathematical model based on Monod growth kinetics was used to predict bacteriocin production and showed results consistent with conventional treatment data. MEF did not have a significant effect on the lag time, maximum specific growth rate, biomass production and pH change under the different experimental conditions at each specific temperature. Based on the observations, bacteriocin activity under the presence of MEF at the early stage of fermentation increased without significant change in the final biomass. PMID- 17461423 TI - Production of lentiviral vectors by large-scale transient transfection of suspension cultures and affinity chromatography purification. AB - The use of lentiviral vectors as gene delivery vehicles has become increasingly popular in recent years. The growing interest in these vectors has created a strong demand for large volumes of vector stocks, which entails the need for scaleable vector manufacturing procedures. In this work, we present a simple and robust process for the production of lentiviral vectors using scaleable production and purification methodologies. Lentivirus particles were produced by transient transfection of serum-free suspension-growing 293 EBNA-1 cells with four plasmids encoding the vector components using linear polyethylenimine (PEI) as transfection reagent. This process was successfully scaled-up from shake flasks to a 3-L bioreactor from which 10(10) IVP were recovered. In addition, an affinity chromatography protocol designed for purification of bioactive oncoretroviral vectors has been adapted in this work for the purification of VSV G pseudotyped lentiviral vectors. Using heparin affinity chromatography, lentiviral particles were concentrated and purified directly from the clarified supernatants. During this step, a recovery of 53% of infective lentiviral particles was achieved while removing 94% of the impurities contained in the supernatant. PMID- 17461424 TI - Size comparison between proteins PEGylated with branched and linear poly(ethylene glycol) molecules. AB - Therapeutic proteins conjugated with branched poly(ethylene glycol) (PEG) have extended in vivo circulation half-lives compared to linear PEG-proteins, thought to be due partly to a greater hydrodynamic volume of branched PEG-proteins, which reduces the glomerular sieving coefficient. In this paper, viscosity radii of PEGylated alpha-lactalbumin (M(r) = 14.2 kDa) and bovine serum albumin (M(r) = 67 kDa) prepared with linear and branched PEGs (with nominal molecular weights 5, 10, 20 and 40 kDa) were compared experimentally using size exclusion chromatography (SEC). PEG adduct:protein molecular weight ratios of the PEGylated proteins covered the range 1:12 to 6:1. Direct comparisons of experimentally measured viscosity radii were found to be misleading due to differences between actual and nominal molecular weights of the PEG reagents used. Comparison with predicted viscosity radii shows that there is no significant difference between the viscosity radii of branched and linear PEG-proteins having the same total molecular weight of PEG adducts. Therefore, longer in vivo circulation half-lives of branched PEG-proteins compared to linear PEG-proteins are not explained by size difference. It is also calculated that the molecular size cut-off for glomerular filtration, 60 A for a 30 kDa PEG, matches the 30-50 A size range for the pores of the glomerular basement membrane. Finally, it is confirmed that prediction of PEG-protein viscosity radii should be based upon conservation of the total PEG adduct surface area to volume ratio for both linear and branched PEG-proteins regardless of PEGylation extent. PMID- 17461425 TI - A novel solid-liquid two-phase partitioning bioreactor for the enhanced bioproduction of 3-methylcatechol. AB - The bioproduction of 3-methylcatechol from toluene via Pseudomonas putida MC2 was performed in a solid-liquid two-phase partitioning bioreactor with the intent of increasing yield and productivity over a single-phase system. The solid phase consisted of HYTREL, a thermoplastic polymer that was shown to possess superior affinity for the inhibitory 3-methylcatechol compared to other candidate polymers as well as a number of immiscible organic solvents. Operation of a solid-liquid biotransformation utilizing a 10% (w/w) solid (polymer beads) to liquid phase ratio resulted in the bioproduction of 3-methylcatechol at a rate of 350 mg/L-h, which compares favorably to the single phase productivity of 128 mg/L-h. . HYTREL polymer beads were also reconstituted into polymer sheets, which were placed around the interior circumference of the bioreactor and successfully removed 3 methylcatechol from solution resulting in a rate of 3-methylcatechol production of 343 mg/L-h. Finally, a continuous biotransformation was performed in which culture medium was circulated upwards through an external extraction column containing HYTREL beads. The design maintained sub lethal concentrations of 3 methylcatechol within the bioreactor by absorbing produced 3-methylcatechol into the polymer beads. As 3-methylcatechol concentrations in the aqueous phase approached 500 mg/L the extraction column was replaced (twice) with a fresh column and the process was continued representing a simple and effective approach for the continuous bioproduction of 3-methylcatechol. Recovery of 3 methylcatechol from HYTREL was also achieved by bead desorption into methanol. PMID- 17461426 TI - Elucidation of the mechanism and end products of glutaraldehyde crosslinking reaction by X-ray structure analysis. AB - Glutaraldehyde has been used for several decades as an effective crosslinking agent for many applications including sample fixation for microscopy, enzyme and cell immobilization, and stabilization of protein crystals. Despite of its common use as a crosslinking agent, the mechanism and chemistry involved in glutaraldehyde crosslinking reaction is not yet fully understood. Here we describe feasibility study and results obtained from a new approach to investigate the process of protein crystals stabilization by glutaraldehyde crosslinking. It involves exposure of a model protein crystal (Lysozyme) to glutaraldehyde in alkaline or acidic pH for different incubation periods and reaction arrest by medium exchange with crystallization medium to remove unbound glutaraldehyde. The crystals were subsequently incubated in diluted buffer affecting dissolution of un-crosslinked crystals. Samples from the resulting solution were subjected to protein composition analysis by gel electrophoresis and mass spectroscopy while crosslinked, dissolution resistant crystals were subjected to high resolution X-ray structural analysis. Data from gel electrophoresis indicated that the crosslinking process starts at specific preferable crosslinking site by lysozyme dimer formation, for both acidic and alkaline pH values. These dimer formations were followed by trimer and tetramer formations leading eventually to dissolution resistant crystals. The crosslinking initiation site and the end products obtained from glutaraldehyde crosslinking in both pH ranges resulted from reactions between lysine residues of neighboring protein molecules and the polymeric form of glutaraldehyde. Reaction rate was much faster at alkaline pH. Different reaction end products, indicating different reaction mechanisms, were identified for crosslinking taking place under alkaline or acidic conditions. PMID- 17461427 TI - Proteome analysis of antibody-producing CHO cell lines with different metabolic profiles. AB - Two-dimensional gel electrophoresis and tandem mass spectrometry were used to identify proteins associated with a metabolic shift during fed-batch cultures of two recombinant antibody-producing CHO cell lines. The first cell line underwent a marked change in lactate metabolism during culture, initially producing lactate and then consuming it, while the second cell line produced lactate for a similar duration but did not later consume it. The first cell line displayed a declining specific antibody productivity during culture, correlating to the 2-D gel results and the intracellular antibody concentration determined by HPLC. Several statistical analysis methods were compared during this work, including a fixed fold-change criterion and t-tests using standard deviations determined in several ways from the raw data and mathematically transformed data. Application of a variance-stabilizing transformation enabled the use of a global empirical standard deviation in the t-tests. Most of the protein spots changing in each cell line did not change significantly in the other cell line. A substantial fraction of the changing proteins were glycolytic enzymes; others included proteins related to antibody production, protein processing, and cell structure. Enolase, pyruvate kinase, BiP/GRP78, and protein disulfide isomerase were found in spots that changed over time in both cell lines, and some protein changes differed from previous reports. These data provide a foundation for future investigation of metabolism in industrially relevant mammalian cell culture processes, and suggest that along with differences between cell types, the proteins expressed in cultures with low lactate concentrations may depend on how those conditions were generated. PMID- 17461428 TI - Activity of human P450 2D6 in biphasic solvent systems. AB - Several limitations have restricted the use of P450 enzymes in synthesis, including the narrow substrate specificity of some P450 isoforms, the need for a redox partner and an expensive cofactor, incompatibility with organic solvents, and poor stability. We previously demonstrated that the natural redox partner and cofactor of the promiscuous P450s 3A4 and 2D6 can be efficiently substituted with some cheap hydrogen peroxide donors or organic peroxides. We report here that P450 2D6 maintains as much as 76% of its activity when used in buffer/organic emulsions. Product formation in biphasic solvent systems is comparable whether the natural redox partner and cofactor are used, or a surrogate. As reported for other enzymes, a correlation is observed between the logP and the suitability of a solvent for enzymatic activity. Moreover, the utility of our system was established by demonstrating the transformation of a novel hydrophobic substrate, not modified by P450 2D6 in the absence of organic solvent. PMID- 17461429 TI - Colorimetric detection of mercuric ion (Hg2+) in aqueous media using DNA functionalized gold nanoparticles. PMID- 17461430 TI - Prognostic value of serum liver enzymes levels in type 2 diabetic patients. AB - BACKGROUND: Elevated liver enzymes are associated with cardiovascular disease, while their relationship with cancer-related mortality has not been assessed so far in diabetic patients. METHODS: An observational cohort study was performed on a consecutive series of 1952 type 2 diabetic patients. The association of liver enzymes with all-cause and cause-specific mortality was assessed. Information on all-cause mortality was obtained by the City of Florence Registry Office. RESULTS: The average duration of follow-up was 6.4 +/- 2.7 years. Over that period, 362 deaths were recorded (26.9%), with a yearly mortality rate of 4.2%. Age- and sex-adjusted HR of all-cause mortality for gamma glutamyl-transpeptidase (gamma-GT) gamma-GT > 40 U/l was 1.610 [1.245-2.082]. An increased cardiovascular mortality rate was observed in patients with elevation of gamma-GT, and gamma-GT and/or alanine aminotransferase (ALT), when compared with the rest of the sample (15.3 vs 10.8%, p < 0.05; and 15.2 vs 10.7%, p < 0.05, respectively). Similar results were observed when considering cancer-related mortality. The association of higher gamma-GT levels with all-cause, cardiovascular, and cancer-related mortality was confirmed at a multivariate analysis after adjustment for potential confounders. CONCLUSIONS: The present study shows that, in type 2 diabetic patients, higher gamma-GT, but not ALT, is associated with increased mortality for cardiovascular disease and malignancies. PMID- 17461431 TI - Insulin VNTR I/III genotype is associated with autoantibodies against glutamic acid decarboxylase in newly diagnosed type 1 diabetes. AB - BACKGROUND: In type 1 diabetes (T1D), the influence of age at diagnosis and of the IDDM1 and IDDM2 genetic susceptibility loci on the profile of beta-cell autoantibodies has been demonstrated. We studied these associations in a group of 92 patients (children, adolescents and adults, aged 2-62 years) with newly diagnosed T1D. METHODS: The prevalence of the HLA-DQB1*02 and *0302 alleles and of the classes of variable number of tandem repeats (VNTR) of the insulin gene (INS), and of beta-cell autoantibodies (GADA, IA-2A, ICA and IAA) was determined. Statistical analysis was performed using linear and logistic regression models. RESULTS: The presence of IAA, IA-2A and ICA, but not of GADA, was negatively associated with age at diagnosis. Younger patients were more likely to have multiple autoantibodies. There was a tendency of a higher prevalence of IAA in patients with the HLA-DQB1*02/0302 genotype or with the DQB1*0302 allele compared to patients lacking these markers. As a novel observation, the INS VNTR I/III genotype was significantly associated with the presence of GADA (OR = 4.79; p = 0.018). CONCLUSION: The association between the INS VNTR I/III genotype and GADA may suggest that in patients with T1D lacking the INS VNTR I/I genotype, the effect of other susceptibility factors prevails, which promotes the development of autoimmunity to beta-cell antigens other than insulin. PMID- 17461432 TI - The protective effect of N-acetylcysteine against cyclosporine A-induced hepatotoxicity in rats. AB - The immunosuppressive agent cyclosporine A (CsA) has been reported to exert measurable hepatotoxic effects. One of the causes leading to hepatotoxicity is thought to be reactive oxygen radical formation. The aim of this study was to investigate the effects of N-acetylcysteine (NAC) treatment on CsA-induced hepatic damage by both analysing superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), aspartate aminotransferase (AST) and alanine transaminase (ALT) activities with malondialdehyde (MDA) and nitric oxide (NO) levels, and using an histological approach. CsA administration produced a decrease in hepatic SOD activity, and co-administration of NAC with CsA resulted in an increase in SOD activity. MDA and NO levels increased in the CsA group and NAC treatment prevented those increases. A significant elevation in serum AST and ALT activities was observed in the CsA group, and when NAC and CsA were co administered, the activities of AST and ALT were close to the control levels. CsA treatment caused evident morphological alterations. Control rats showed no abnormality in the cytoarchitecture of the hepatic parenchyma. The co administration of NAC with CsA showed no signs of alteration and the morphological pattern was almost similar to the control group. In conclusion, CsA induced liver injury and NAC treatment prevented the toxic side effects induced by CsA administration through the antioxidant and radical scavenging effects of NAC. PMID- 17461433 TI - Isolation and endotoxin activities of lipopolysaccharides from cyanobacterial cultures and complex water blooms and comparison with the effects of heterotrophic bacteria and green alga. AB - Massive cyanobacterial water blooms are serious environmental and health problems worldwide. While some cyanobacterial toxins such as peptide microcystins have been investigated extensively, other toxic components of cyanobacteria (e.g. lipopolysaccharides, LPS) are poorly understood. The present study characterized endotoxin activities of LPS isolated from (i) laboratory cyanobacterial cultures, (ii) cyanobacterial water bloom samples dominated by Microcystis sp., Planktothrix sp., Aphanizomenon sp. and Anabaena sp., (iii) heterotrophic Gram negative bacteria Escherichia coli, Kluyvera intermedia, Pseudomonas putida and Pseudomonas fluorescens and (iv) green alga Pseudokirchneriella subcapitata. Toxicity results derived with Limulus amebocyte lysate assay (LAL-test) showed that endotoxin activities of LPS from both cyanobacteria and heterotrophic bacteria were comparable and the values were within a similar range (1 x 10(3)-1 x 10(6) Endotoxin Units, EU, per mg of isolated LPS). The highest activities among the cyanobacterial samples were observed in the Aphanizomenon sp. dominated water bloom. The results also suggest generally higher endotoxin activities in complex natural samples than in laboratory cyanobacterial cultures. Further, experiments with the eukaryotic green alga P. subcapitata demonstrated a need for careful purification of the LPS extracts prior to testing with the LAL assay as several contaminants may overestimate endotoxin activities. This study shows relatively high pyrogenicity of LPS from various cyanobacteria. Further research should focus on detailed toxicological and ecotoxicological characterization of LPS in massive cyanobacterial water blooms. PMID- 17461434 TI - Mitochondria a key role in microcystin-LR kidney intoxication. AB - Microcystins (MCs) are a group of closely related cyclic heptapeptides produced by a variety of common cyanobacteria. These toxins have been implicated in both human and livestock mortality. Microcystin-LR could affect renal physiology by altering vascular, glomerular and urinary parameters, indicating that MC-LR could act directly on the kidney. The aim of the current work was to examine the effect of MC-LR on mitochondrial oxidative phosphorylation of rat kidney isolated mitochondria.Furthermore, microcystin-LR decreased both state 3 and carbonylcyanide p-trifluoromethoxyphenylhydrazone (FCCP)-uncoupled respiration. The transmembrane potential was strongly depressed by MC-LR in a concentration dependent manner, pointing to an uncoupling effect; however, microcystin-LR did not increase the permeability of the inner mitochondria membrane to protons. Therefore, the transmembrane decrease was a consequence of a strong inhibitory effect on redox complexes. The addition of uncoupling concentrations of MC-LR to Ca(2+)-loaded mitochondria treated with ruthenium red resulted in mitochondrial permeability transition pore (MPTP) opening, as evidenced by mitochondrial swelling in isosmotic sucrose medium. Mitochondrial swelling in the presence of Ca(2+) was prevented by cyclosporin A and was drastically inhibited by catalase and dithiothreitol, indicating the participation of mitochondrial generated reactive oxygen species in this process. From this study it can be concluded that the bioenergetic lesion promoted by microcystin-LR seems to be sufficient to explain renal injury. PMID- 17461435 TI - Clinical experience of the use of a pharmacological treatment algorithm for major depressive disorder in patients with advanced cancer. AB - The objective of this study was to describe the applicability and the dropout of the pharmacological treatment algorithm for major depressive disorder in patients with advanced cancer. Psychiatrists treated major depressive disorder in advanced cancer patients on the basis of the algorithm. For discussing the problems related to the algorithm, we reviewed the reasons for the non-application of the algorithm and the reasons for dropout of patients within a week of initiation of treatment. The algorithm was applied in 54 of 59 cases (applicability rate, 92%). The reasons for the non-application of the algorithm were as follows: the need to add a benzodiazepine to an antidepressant in 4 cases and the need to choose alprazolam despite the depression being moderate in severity, in order to obtain a rapid onset action and reduce anxiety in a patient with short prognosis. Nineteen of the 55 patients dropped out within a week of initiation of treatment based on the algorithm. Delirium was the most frequent reason for dropout. The applicability rate was high, but several problems were identified, including those related to the combination of antidepressants and benzodiazepines, pharmacological treatment of depression in patients with short prognosis, and delirium due to antidepressants. PMID- 17461436 TI - Intracellular water-specific MR of microbead-adherent cells: the HeLa cell intracellular water exchange lifetime. AB - Quantitative characterization of the intracellular water (1)H MR signal from cultured cells will provide critical biophysical insight into the MR signal from tissues in vivo. Microbeads provide a robust immobilization substrate for the many mammalian cell lines that adhere to surfaces and also provide sufficient cell density for observation of the intracellular water MR signal. However, selective observation of the intracellular water MR signal from perfused, microbead-adherent mammalian cells requires highly effective suppression of the extracellular water MR signal. We describe how high-velocity perfusion of microbead-adherent cells results in short apparent (1)H MR longitudinal and transverse relaxation times for the extracellular water in a thin slice selected orthogonal to the direction of flow. When combined with a spin-echo pulse sequence, this phenomenon provides highly effective suppression of the extracellular water MR signal. This new method is exploited here to quantify the kinetics of water exchange from the intracellular to extracellular spaces of HeLa cells. The time constant describing water exchange from intracellular to extracellular spaces, also known as the exchange lifetime for intracellular water, is 119 +/- 14 ms. PMID- 17461438 TI - Microarray-based analysis for hepatocellular carcinoma: from gene expression profiling to new challenges. AB - Accumulation of mutations and alterations in the expression of various genes result in carcinogenesis, and the development of microarray technology has enabled us to identify the comprehensive gene expression alterations in oncogenesis. Many studies have applied this technology for hepatocellular carcinoma (HCC), and identified a number of candidate genes useful as biomarkers in cancer staging, prediction of recurrence and prognosis, and treatment selection. Some of these target molecules have been used to develop new serum diagnostic markers and therapeutic targets against HCC to benefit patients. Previously, we compared gene expression profiling data with classification based on clinicopathological features, such as hepatitis viral infection or liver cancer progression. The next era of gene expression analysis will require systematic integration of expression profiles with other types of biological information, such as genomic locus, gene function, and sequence information. We have reported integration between expression profiles and locus information, which is effective in detecting structural genomic abnormalities, such as chromosomal gains and losses, in which we showed that gene expression profiles are subject to chromosomal bias. Furthermore, array-based comparative genomic hybridization analysis and allelic dosage analysis using genotyping arrays for HCC were also reviewed, with comparison of conventional methods. PMID- 17461439 TI - Spleen-preserving distal pancreatectomy with conservation of the splenic artery and vein. AB - Preservation of the spleen at distal pancreatectomy has recently attracted considerable attention. Since our first successful trial, spleen-preserving distal pancreatectomy with conservation of the splenic artery and vein for tumors of the pancreas and chronic pancreatitis has been performed more frequently. The technique for spleen-preserving distal pancreatectomy with conservation of the splenic artery and vein are outlined. The splenic vein is identified behind the pancreas and within the thin connective tissue membrane. The connective tissue membrane is cut longitudinally above the splenic vein. An important issue is to remove the splenic vein from the body of the pancreas toward the spleen, since a different approach may be very difficult. The pancreas is preferably removed from the splenic artery toward the head of the pancreas itself. This procedure is much easier than removing the pancreas from the vein side. One patient had undergone distal gastrectomy for duodenal ulcer, with reconstruction by Billroth II technique. If distal pancreatectomy with splenectomy had been performed for the lesion of the distal pancreas at the time, the residual stomach would also have to be resected. The potential damage done to the patient by reconstruction of the gastrointestinal tract in combination with distal pancreatectomy and splenectomy would have been much greater than with distal pancreatectomy only with preservation of the spleen and residual stomach. Benign lesions as well as low grade malignancy of the body and tail of the pancreas may be a possible indication for this procedure. PMID- 17461437 TI - Intestinal M cells: the fallible sentinels? AB - The gastrointestinal tract represents the largest mucosal membrane surface in the human body. The immune system in the gut is the first line of host defense against mucosal microbial pathogens and it plays a crucial role in maintaining mucosal homeostasis. Membranous or microfold cells, commonly referred to as microfold cells, are specialized epithelial cells of the gut-associated lymphoid tissues (GALT) and they play a sentinel role for the intestinal immune system by delivering luminal antigens through the follicle-associated epithelium to the underlying immune cells. M cells sample and uptake antigens at their apical membrane, encase them in vesicles to transport them to the basolateral membrane of M cells, and from there deliver antigens to the nearby lymphocytes. On the flip side, some intestinal pathogens exploit M cells as their portal of entry to invade the host and cause infections. In this article, we briefly review our current knowledge on the morphology, development, and function of M cells, with an emphasis on their dual role in the pathogenesis of gut infection and in the development of host mucosal immunity. PMID- 17461440 TI - Multi-disciplinary treatment for cholangiocellular carcinoma. AB - Cholangiocarcinoma (CC) is rare malignant tumors composed of cells that resemble those of the biliary tract. It is notoriously difficult to diagnose, and is associated with a high mortality. Traditionally, CC is divided into intrahepatic and extrahepatic disease according to its location within the biliary tree. Intrahepatic cholangiocellular carcinoma (IH-CCC) or peripheral cholangiocellular carcinoma (CCC) appears within the second bifurcation of hepatic bile duct, and is the second most common primary liver cancer following hepatocellular carcinoma (HCC), IH-CCC or peripheral CCC often presents with advanced clinical features, and the cause for this cancer rise is still unclear. MRI, CT and PET provide useful diagnostic information in those patients. Surgical resection is the only chance for cure, with results depending on selected patients and careful surgical technique. Liver transplantation could offer long-term survival in selected patients when combined with chemotherapy. Chemotherapy, radiation therapy or combination therapies remain as the only treatment for inoperable patients. However, these are uniformly ineffective in patients' survival. PMID- 17461441 TI - Current surgical treatment for bile duct cancer. AB - Since extrahepatic bile duct cancer is difficult to diagnose and to cure, a safe and radical surgical strategy is needed. In this review, the modes of infiltration and spread of extrahepatic bile duct cancer and surgical strategy are discussed. Extended hemihepatectomy, with or without pancreatoduodenectomy (PD), plus extrahepatic bile duct resection and regional lymphadenectomy has recently been recognized as the standard curative treatment for hilar bile duct cancer. On the other hand, PD is the choice of treatment for middle and distal bile duct cancer. Major hepatectomy concomitant with PD (hepatopancreatoduodenectomy) has been applied to selected patients with widespread tumors. Preoperative biliary drainage (BD) followed by portal vein embolization (PVE) enables major hepatectomy in patients with hilar bile duct cancer without mortality. BD should be performed considering the surgical procedure, especially, in patients with separated intrahepatic bile ducts caused by hilar bile duct cancer. Right or left trisectoriectomy are indicated according to the tumor spread and biliary anatomy. As a result, extended radical resection offers a chance for cure of hilar bile duct cancer with improved resectability, curability, and a 5-year survival rate of 40%. A 5-year survival rate has ranged from 24% to 39% after PD for middle and distal bile duct cancer. PMID- 17461442 TI - Artificial and bioartificial liver support: a review of perfusion treatment for hepatic failure patients. AB - Liver transplantation and blood purification therapy, including plasmapheresis, hemodiafiltration, and bioartificial liver support, are the available treatments for patients with severe hepatic failure. Bioartificial liver support, in which living liver tissue is used to support hepatic function, has been anticipated as an effective treatment for hepatic failure. The two mainstream systems developed for bioartificial liver support are extracorporeal whole liver perfusion (ECLP) and bioreactor systems. Comparing various types of bioartificial liver in view of function, safety, and operability, we concluded that the best efficacy can be provided by the ECLP system. Moreover, in our subsequent experiments comparing ECLP and apheresis therapy, ECLP offers more ammonia metabolism than HD and HF. In addition, ECLP can compensate amino acid imbalance and can secret bile. A controversial point with ECLP is the procedure is labor intensive, resulting in high costs. However, ECLP has the potential to reduce elevated serum ammonia levels of hepatic coma patients in a short duration. When these problems are solved, bioartificial liver support, especially ECLP, can be adopted as an option in ordinary clinical therapy to treat patients with hepatic failure. PMID- 17461443 TI - Rising costs and hospital admissions for hepatocellular carcinoma in Portugal (1993-2005). AB - AIM: To determine, for hepatocellular carcinoma (HCC), the patient demographic profile and costs of their admissions to the hospitals of the Portuguese National Health System from 1993 to 2005. METHODS: The National Registry (ICD-9CM, Inter national Classification of Diseases, 155.0) provided data from the 97 Hospitals in Portugal. RESULTS: We studied 7932 admissions that progressively rose from 292 in 1993 to 834 in 2005, having a male predominance of 78% (6130/7932). The global rate of hospital admissions for HCC rose from 3.1/10(5) in 1993 to 8.3/10(5) in 2005. The average length of stay decreased from 17.5 +/- 17.9 d in 1993 to 9.3 +/ 10.4 d in 2005, P < 0.001. The average hospital mortality for HCC remained high over these years, 22.3% in 1993 and 26.7% in 2005. Nationally, hospital costs (in Euros - EUR) rose in all variables studied: overall costs from 533,000 Euros in 1993, to 462,9000 Euros in 2005, cost per day of stay from 105 Euros in 1993, to 597 Euros in 2005, average cost of each admission from 1828 Euros in 1993, to 5550 Euros in 2005. In 2005, 1.8% (15/834) of hospital admissions for HCC were related to liver transplant, and responsible for a cost of about 1.5 million Euros, corresponding to one third of the overall costs for HCC admissions in that same year. CONCLUSION: From 1993 to 2005 hospital admissions in Portugal for HCC tripled. Overall costs for these admissions increased 9 times, with all variables related to cost analysis rising accordingly. Liver transplant, indicated in a small group of patients, showed a disproportionate increase in costs. PMID- 17461444 TI - Stool-based DNA testing, a new noninvasive method for colorectal cancer screening, the first report from Iran. AB - AIM: To detect tumor-associated DNA changes in stool samples among Iranian patients with colorectal cancer (CRC) compared to healthy individuals using BAT 26, p16 hypermethylation and long DNA markers. METHODS: Stool DNA was isolated from 45 subjects including 25 CRC patients and 20 healthy individuals using a new, fast and easy extraction method. Long DNA associated with tumor was detected using polymerase chain reaction method. Microsatellite studies were performed utilizing denaturating polyacrylamide gel to determine the instability of BAT-26. Methylation status of p16 promoter was analyzed using methylation-specific PCR (MSP). RESULTS: The results showed a significant difference in existence of long DNA (16 in patients vs 1 in controls, P < 0.001) and p16 (5 in patients vs none in controls, P = 0.043) in the stool samples of two groups. Long DNA was detected in 64% of CRC patients; whereas just one of the healthy individuals was positive for Long DNA. p16 methylation was found in 20% of patients and in none of healthy individuals. Instability of BAT-26 was not detected in any of stool samples. CONCLUSION: We could detect colorectal cancer related genetic alterations by analyzing stool DNA with a sensitivity of 64% and 20% and a specificity of 95% and 100% for Long DNA and p16 respectively. A non-invasive molecular stool-based DNA testing can provide a screening strategy in high-risk individuals. However, additional testing on more samples is necessary from Iranian subjects to determine the exact specificity and sensitivity of these markers. PMID- 17461445 TI - Rosiglitazone enhances fluorouracil-induced apoptosis of HT-29 cells by activating peroxisome proliferator-activated receptor gamma. AB - AIM: To examine whether and how rosiglitazone enhances apoptosis induced by fluorouracil in human colon cancer (HT-29) cells. METHODS: Human colon cancer HT 29 cells were cultured in vitro and treated with fluorouracil and/or rosiglitazone. Proliferation and growth of HT-29 cells were evaluated by MTT assay and trypan blue exclusion methods, respectively. The apoptosis of HT-29 cells was determined by acridine orange/ethidium bromide staining and flow cytometry using PI fluorescence staining. The expressions of peroxisome proliferator-activated receptor gamma (PPARgamma), Bcl-2 and Bax in HT-29 cells were analyzed by Western blot. RESULTS: Although rosiglitazone at the concentration below 30 micromol/L for 72 h exerted almost no inhibitory effect on proliferation and growth of HT-29 cells, it could significantly enhance fluorouracil-induced HT-29 cell proliferation and growth inhibition. Furthermore, 10 micromol/L rosilitazone did not induce apoptosis of HT-29 cells but dramatically enhanced fluorouracil-induced apoptosis of HT-29 cells. However, rosiglitazone did not improve apoptosis induced by fluorouracil in HT-29 cells pretreated with GW9662, a PPARgamma antagonist. Meanwhile, the expression of Bax and PPARgamma was up-regulated, while the expression of Bcl-2 was down regulated in HT-29 cells treated with rosiglitazone in a time-dependent manner. However, the effect of rosiglitazone on Bcl-2 and Bax was blocked or diminished in the presence of GW9662. CONCLUSION: Rosiglitazone enhances fluorouracil-induced apoptosis of HT-29 cells by activating PPARgamma. PMID- 17461446 TI - Expression of mutant type-p53 products in H pylori-associated chronic gastritis. AB - AIM: To investigate the mutation of p53 immunohistochemically in non-tumorous gastric mucosa with H pylori infection before and after H pylori eradication therapy. METHODS: 53 subjects (36 male, 17 female, mean age +/- SEM, 57.1 +/- 12.1) undergoing endoscopic examination were included in this study. 42 of 53 patients were H pylori-positive, and 11 were H pylori-negative. All H pylori positive patients had successful eradication therapy. Biopsy specimens were taken from five points of the stomach, as recommended by the updated Sydney system. Immunohistochemical studies were performed by using primary antibodies against p53 (DO-7 and PAb240). RESULTS: p53 (DO-7 and PAb240) immunoreactivity was shown in the neck region of the gastric pits, however, quite a few cells were found to be immunopositive for p53 (PAb240) in the H pylori-infected gastric mucosa. The proportion of patients immunopositive for p53 (PAb240) was significantly reduced 6 mo after eradication [28/42 (66.7%) to 6/42 (14.3%)] (P < 0.05), while the biopsies taken from H pylori-negative patients showed no immunoreactivity for p53 (PAb240). p53 (PAb240)-positive patients were divided into two groups by the number of positive cells detected: one with more than six positive cells per 10 gastric pits (group A, n = 12), and the other with less than five positive cells per 10 gastric pits (group B, n = 30). Atrophy scores in group A were significant higher than those in group B at the greater curvature of the antrum (group A: 2.00 +/- 0.14 vs group B: 1.40 +/- 0.15, P = 0.012), the lesser curvature of the corpus (group A: 2.00 +/- 0.21 vs group B: 1.07 +/- 0.23, P = 0.017), and the greater curvature of the corpus (group A: 1.20 +/- 0.30 vs group B: 0.47 +/- 0.21, P = 0.031). Group A showed significant higher intestinal metaplasia scores than group B only at the lesser curvature of the antrum (group A: 2.10 +/- 0.41 vs group B: 1.12 +/- 0.29, P = 0.035). CONCLUSION: H pylori-associated chronic gastritis expressed the mutant-type p53, which was significantly associated with more severe atrophic and metaplastic changes. H pylori eradication led to a significant reduction in the expression of the mutant-type p53. It is considered that H pylori-infected chronic gastritis is associated with a genetic instability that leads to gastric carcinogenesis, and H pylori eradication may prevent gastric cancer. PMID- 17461447 TI - Th immune response induced by H pylori vaccine with chitosan as adjuvant and its relation to immune protection. AB - AIM: To study the immunological protective effect of H pylori vaccine with chitosan as an adjuvant and its mechanism. METHODS: Female BALB/c mice were randomly divided into seven groups and orally immunized respectively with PBS, chitosan solution, chitosan particles, H pylori antigen, H pylori antigen plus cholera toxin (CT), H pylori antigen plus chitosan solution, H pylori antigen plus chitosan particles once a week for four weeks. Four weeks after the last immunization, the mice were challenged twice by alive H pylori (1 x 10(9) CFU/mL) and sacrificed. Part of the gastric mucosa was embedded in paraffin, cut into sections and assayed with Giemsa staining. Part of the gastric mucosa was used to quantitatively culture H pylori. ELISA was used to detect cytokine level in gastric mucosa and anti- H pylori IgG1, IgG2a levels in serum. RESULTS: In the groups with chitosan as an adjuvant, immunological protection was achieved in 60% mice, which was significantly higher than in groups with H pylori antigen alone and without H pylori antigen (P < 0.05 or 0.001). Before challenge, the level of IFN and IL-12 in gastric mucosa was significantly higher in the groups with chitosan as an adjuvant than in the control group and the group without adjuvant (P < 0.05 or 0.005). After challenge, the level of IFN and IL-12 was significantly higher in the groups with adjuvant than in the groups without adjuvant and antigen (P < 0.05 or 0.001). Before challenge, the level of IL-2 in gastric mucosa was not different among different groups. After challenge the level of IL-2 was significantly higher in the groups with adjuvant than in the control group (P < 0.05 or 0.001). Before challenge, the level of IL-10 in gastric mucosa was significantly higher in the groups with chitosan as an adjuvant than in other groups without adjuvant (P < 0.05 or 0.01). After challenge, the level of IL-10 was not different among different groups. Before challenge, the level of IL-4 in gastric mucosa was significantly higher in the groups with chitosan as an adjuvant than in other groups without adjuvant (P < 0.05). After challenge, the level of IL-4 was significantly higher in the groups with chitosan particles as an adjuvant than in the group with CT as an adjuvant (P < 0.05), and in the group with chitosan solution as an adjuvant, the level of IL-4 was significantly higher than that in control group, non-adjuvant group and the groups with CT (P < 0.05 or 0.001). The ratio of anti- H pylori IgG2a/IgG1 in serum was significantly lower in the groups with chitosan as an adjuvant than in the groups with CT as an adjuvant or without adjuvant (P < 0.01). CONCLUSION: H pylori vaccine with chitosan as an adjuvant can protect against H pylori infection and induce both Th1 and Th2 type immune response. PMID- 17461448 TI - Epithelial cells with hepatobiliary phenotype: is it another stem cell candidate for healthy adult human liver? AB - AIM: To investigate the presence and role of liver epithelial cells in the healthy human adult liver. METHODS: Fifteen days after human hepatocyte primary culture, epithelial like cells emerged and started proliferating. Cell colonies were isolated and subcultured for more than 160 d under specific culture conditions. Cells were analyzed for each passage using immunofluorescence, flow cytometry and reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: Flow cytometry analysis demonstrated that liver epithelial cells expressed common markers for hepatic and stem cells such as CD90, CD44 and CD29 but were negative for CD34 and CD117. Using immunofluorescence we demonstrated that liver epithelial cells expressed not only immature (alpha-fetoprotein) but also differentiated hepatocyte (albumin and CK-18) and biliary markers (CK-7 and 19), whereas they were negative for OV-6. RT-PCR analysis confirmed immunofluorescence data and revealed that liver epithelial cells did not express mature hepatocyte markers such as CYP2B6, CYP3A4 and tyrosine amino-transferase. Purified liver epithelial cells were transplanted into SCID mice. One month after transplantation, albumin positive cell foci were detected in the recipient mouse parenchyma. CONCLUSION: According to their immature and bipotential phenotype, liver epithelial cells might represent a pool of precursors in the healthy human adult liver other than oval cells. PMID- 17461449 TI - Roles of the MEK1/2 and AKT pathways in CXCL12/CXCR4 induced cholangiocarcinoma cell invasion. AB - AIM: To evaluate the expression of C-X-C motif chemokine receptor 4 (CXCR4) and its signaling cascades, which were previously identified as a key factor for cancer cell progression and metastasis, in cholangiocarcinoma cell lines. METHODS: The expression of CXCR4 and its signaling cascades were determined in the cholangiocarcinoma cell lines (RMCCA1 and KKU100) by Western blotting. The invasion assays and the detection of actin polymerization were tested in these cholangiocarcinoma cells treated with CXC chemokine ligand -12 (CXCL12). RESULTS: Expression of CXCR4 was detected in both cholangiocarcinoma cell lines and activation of CXCR4 with CXCL12 triggered the signaling via the extracellular signal-regulated kinase-1/2 (ERK1/2) and phosphoinositide 3-kinase (PI3K) and induction of cholangiocarcinoma cell invasion, and displayed high levels of actin polymerization. Addition of CXCR4 inhibitor (AMD3100) abrogated CXCL12-induced phosphorylation of MEK1/2 and Akt in these cells. Moreover, treatment with MEK1/2 inhibitor (U0126) or PI3K inhibitor (LY294002) also attenuated the effect of CXCL12-induced cholangiocarcinoma cell invasion. CONCLUSION: These results indicated that the activation of CXCR4 and its signaling pathways (MEK1/2 and Akt) are essential for CXCL12-induced cholangiocarcinoma cell invasion. This rises Implications on a potential role for the inhibition of CXCR4 or its signal cascades in the treatment of cholangiocarcinoma. PMID- 17461450 TI - Usefulness of fecal lactoferrin and hemoglobin in diagnosis of colorectal diseases. AB - AIM: To evaluate prospectively usefulness of fecal lactoferrin (Lf) and fecal hemoglobin (Hb) in the diagnosis of colorectal diseases. METHODS: Fecal Lf and Hb were measured using ELISA in 872 patients before they underwent colorectal endoscopy. RESULTS: Lf was positive in 18 (50%) of 36 patients with colorectal cancer, 25 (15.9%) of 157 with colorectal polyps, 29 (46.8%) of 62 with ulcerative colitis, and 25 (62.5%) of 40 (62.5%) with Crohn's disease. The Hb positive rates were 50%, 12.1%, 41.9% and 32.5%, respectively. Of the 318 patients free of abnormalities by colorectal endoscopy, Lf was positive in 29 (9.1%) and Hb was positive in 15 (4.7%). Among patients with Crohn's disease, the Lf-positive rate was significantly higher than the Hb-positive rate. If either high Lf or Hb levels were considered positive, the positive rates rose to 61.1%, 51.6%, and 67.5% in the colorectal cancer group, ulcerative colitis group, and Crohn's disease group, respectively. If both high Lf and Hb levels were rated positive, the positive predictive values (PPV) were 21% for colorectal cancer, 33% for ulcerative colitis, and 17% for Crohn's disease, and PPV of high Hb level alone was 18%, 25% and 13%, respectively. CONCLUSION: Fecal Lf and Hb were found useful in the detection of colorectal diseases, and the combination of the two measurements appears to increase the sensitivity and efficacy of diagnosis. PMID- 17461451 TI - A new iron free treatment with oral fish cartilage polysaccharide for iron deficiency chronic anemia in inflammatory bowel diseases: a pilot study. AB - AIM: To investigate the effect of a new oral preparation, highly concentrated in fish cartilage, in a group of inflammatory bowel diseases (IBD) patients with chronic iron deficient anemia. METHODS: In an open label pilot study, we supple mented a group of 25 patients (11 with Crohn's disease and 14 with ulcerative colitis) in stable clinical conditions and chronic anemia with a food supplement which does not contain iron but contains a standardized fraction of fish cartilage glycosaminoglycans and a mixture of antioxidants (Captafer Medestea, Turin, Italy). Patients received 500 mg, twice a day during meals, for at least 4 mo. Patients were suggested to maintain their alimentary habit. At time 0 and after 2 and 4 mo, emocrome, sideremia and ferritin were examined. Paired data were analyzed with Student's t test. RESULTS: Three patients relapsed during the study (2 in the 3rd mo, 1 in the 4th mo), two patients were lost to follow up and two patients dropped out (1 for orticaria, 1 for gastric burning). Of the remaining 18 patients, levels of serum iron started to rapidly increase within the 2nd mo of treatment, P < 0.05), whereas serum ferritin and hemoglobin needed a longer period to significantly improve their serum levels (mo 4) P < 0.05. The product was safe, easy to administer and well tolerated by patients. CONCLUSION: These data suggest a potential new treatment for IBD patients with iron deficiency chronic anemia and warrant further larger controlled studies. PMID- 17461452 TI - Nonalcoholic fatty liver disease is a novel predictor of cardiovascular disease. AB - AIM: To clarify whether nonalcoholic fatty liver disease (NAFLD) increases the risk of cardiovascular disease. METHODS: We carried out a prospective observational study with a total of 1637 apparently healthy Japanese men and women who were recruited from a health check-up program. NAFLD was diagnosed by abdominal ultrasonography. The metabolic syndrome (MS) was defined according to the modified National Cholesterol Education Program (NCEP) ATP III criteria. Five years after the baseline evaluations, the incidence of cardiovascular disease was assessed by a self-administered questionnaire. RESULTS: Among 1221 participants available for outcome analyses, the incidence of cardiovascular disease was higher in 231 subjects with NAFLD at baseline (5 coronary heart disease, 6 ischemic stroke, and 1 cerebral hemorrhage) than 990 subjects without NAFLD (3 coronary heart disease, 6 ischemic stroke, and 1 cerebral hemorrhage). Multivariate analyses indicated that NAFLD was a predictor of cardiovascular disease independent of conventional risk factors (odds ratio 4.12, 95% CI, 1.58 to 10.75, P = 0.004). MS was also independently associated with cardiovascular events. But simultaneous inclusion of NAFLD and MS in a multivariate model revealed that NAFLD but not MS retained a statistically significant correlation with cardiovascular disease. CONCLUSION: Although both of them were predictors of cardiovascular disease, NAFLD but not MS retained a statistically significant correlation with cardiovascular disease in a multivariate model. NAFLD is a strong predictor of cardiovascular disease and may play a central role in the cardiovascular risk of MS. PMID- 17461453 TI - Risk factors for Barrett's oesophagus and oesophageal adenocarcinoma: results from the FINBAR study. AB - AIM: To investigate risk factors associated with Barrett's oesophagus and oesophageal adenocarcinoma. METHODS: This all-Ireland population-based case control study recruited 224 Barrett's oesophagus patients, 227 oesophageal adenocarcinoma patients and 260 controls. All participants underwent a structured interview with information obtained about potential lifestyle and environmental risk factors. RESULTS: Gastro-oesophageal reflux was associated with Barrett's [OR 12.0 (95% CI 7.64-18.7)] and oesophageal adenocarcinoma [OR 3.48 (95% CI 2.25 5.41)]. Oesophageal adenocarcinoma patients were more likely than controls to be ex- or current smokers [OR 1.72 (95% CI 1.06-2.81) and OR 4.84 (95% CI 2.72-8.61) respectively] and to have a high body mass index [OR 2.69 (95% CI 1.62-4.46)]. No significant associations were observed between these risk factors and Barrett's oesophagus. Fruit but not vegetables were negatively associated with oesophageal adenocarcinoma [OR 0.50 (95% CI 0.30-0.86)]. CONCLUSION: A high body mass index, a diet low in fruit and cigarette smoking may be involved in the progression from Barrett's oesophagus to oesophageal adenocarcinoma. PMID- 17461454 TI - Influence of H pylori on plasma ghrelin in patients without atrophic gastritis. AB - AIM: To determine the association between H pylori infection and serum ghrelin levels in patients without atrophic gastritis. METHODS: Fifty consecutive patients (24 males and 26 females) with either H pylori-positive gastritis (n = 34) or H pylori-negative gastritis (n = 16) with normal gastric acid secretion determined by 24-h pHmetry and without atrophic gastritis in histopathology were enrolled in this study. Thirty-four H pylori-infected patients were treated with triple therapy consisting of a daily regimen of 30 mg lansoprazole bid, 1 g amoxicillin bid and 500 mg clarithromycin bid for 14 d, followed by an additional 4 wk of 30 mg lansoprazol treatment. H pylori infection was eradicated in 23 of 34 (67.6%) patients. H pylori-positive patients were given eradication therapy. Gastric acidity was determined via intragastric pH catheters. Serum ghrelin was measured by radioimmunoassay (RIA). RESULTS: There was no significant difference in plasma ghrelin levels between H pylori-positive and H pylori-negative groups (81.10 +/- 162.66 ng/L vs 76.51 +/- 122.94 ng/L). In addition, there was no significant difference in plasma ghrelin levels and gastric acidity levels measured before and 3 mo after the eradication therapy. CONCLUSION: H pylori infection does not influence ghrelin secretion in patients with chronic gastritis without atrophic gastritis. PMID- 17461455 TI - Prevalence of coeliac disease in patients with autoimmune thyroiditis in a Turkish population. AB - AIM: To investigate the prevalence of coeliac disease in a series of Turkish patients with autoimmune thyroiditis. METHODS: Sera from 136 consecutive patients with newly diagnosed autoimmune thyroiditis and 119 healthy blood donors were tested for IgA tissue transglutaminase antibody with enzyme-linked immunosorbent assay. Endoscopic mucosal biopsy from the second part of duodenum was performed in patients with positive antibody test. RESULTS: Eight patients (5.9%) and one control subject (0.8%) were positive for IgA tissue transglutaminase antibody (OR: 7.38, 95% CI: 0.91-59.85, P = 0.04). Six patients and one control agreed to take biopsies. Histopathological examination revealed changes classified as Marsh IIIa in one, Marsh II in one, Marsh I in two, and Marsh 0 in two patients with autoimmune throiditis, and Marsh I in one blood donor. CONCLUSION: Turkish patients with autoimmune thyroiditis have an increased risk of coeliac disease and serological screening may be useful for early detection of coeliac disease in these patients. Our findings need to be confirmed in a larger series of patients. PMID- 17461456 TI - Screening and cloning for proteins transactivated by the PS1TP5 protein of hepatitis B virus: a suppression subtractive hybridization study. AB - AIM: To clone and identify human genes transactivated by PS1TP5 by constructing a cDNA subtractive library with suppression subtractive hybridization (SSH) technique. METHODS: SSH and bioinformatics techniques were used for screening and cloning of the target genes transactivated by PS1TP5 protein. The mRNA was isolated from HepG2 cells transfected with pcDNA3.1(-)-myc-his(A)-PS1TP5 and pcDNA3.1(-)-myc-his(A) empty vector, respectively, and SSH technique was employed to analyze the differentially expressed DNA sequence between the two groups. After digestion with restriction enzyme Rsa I, small size cDNAs were obtained. Then tester cDNA was divided into two groups and ligated to the specific adaptor 1 and adaptor 2, respectively. The tester cDNA was hybridized with driver cDNA twice and subjected to nested PCR for two times, and then subcloned into T/A plasmid vectors to set up the subtractive library. Amplification of the library was carried out with E. coli strain DH5alpha. The cDNA was sequenced and analyzed in GenBank with Vector NTI 9.1 and NCBI BLAST software after PCR amplification. RESULTS: The subtractive library of genes transactivated by PS1TP5 was constructed successfully. The amplified library contained 90 positive clones. Colony PCR showed that 70 clones contained 200-1000-bp inserts. Sequence analysis was performed in 30 clones randomly, and the full-length sequences were obtained by bioinformatics technique. Altogether 24 coding sequences were obtained, which consisted of 23 known and 1 unknown. One novel gene with unknown functions was found and named as PS1TP5TP1 after being electronically spliced, and deposited in GenBank (accession number: DQ487761). CONCLUSION: PS1TP5 is closely correlated with immunoregulation, carbohydrate metabolism, signal transduction, formation mechanism of hepatic fibrosis, and occurrence and development of tumor. Understanding PS1TP5 transactive proteins may help to bring some new clues for further studying the biological functions of pre-S1 protein. PMID- 17461457 TI - H pylori exist in the gallbladder mucosa of patients with chronic cholecystitis. AB - AIM: To study whether H pylori locate in the gallbladder mucosa of patients with chronic cholecystitis. METHODS: Using Warthy-Starry (W-S) silver stain and immunohistochemistry stain with anti-H pylori antibodies, we screened paraffin specimens in 524 cases of cholecystitis. H pylori urease gene A (HPUA) and H pylori urease gene B (HPUB) were analyzed by polymerase chain reaction (PCR) in the fresh tissue specimens from 81 cases of cholecystitis. RESULTS: H pylori-like bacteria were found in 13.55% of the gallbladders of the cholecystitis patients using W-S stain. Meanwhile, bacteria positive for H pylori antibodies were also found in 7.1% of the gallbladders of patients with cholecystitis by immunohistochemistry. Of 81 gallbladders, 11 were positive for both HPUA and HPUB, 4 were positive for HPUA only and 7 were positive for HPUB only. CONCLUSION: H pylori exist in the gallbladders of patients with chronic cholecystitis. PMID- 17461458 TI - Clinical and molecular analysis of hereditary non-polyposis colorectal cancer in Chinese colorectal cancer patients. AB - AIM: To analyze the frequency of hereditary non-polyposis colorectal cancer (HNPCC) in Chinese colorectal cancer (CRC) patients, and to discuss the value of microsatellite instability (MSI) and/or immunohistochemistry (IHC) for MSH2/MLH1 protein analysis as pre-screening tests in China. METHODS: The Amsterdam criteria I and II (clinical diagnosis) and/or germline hMLH1/hMSH2 mutations (genetic diagnosis) were used to classify HNPCC families. Genetic tests, including microsatellite instability, immunohistochemistry for MSH2/MLH1 proteins and hMSH2/hMLH1 genes, were performed in each proband. RESULTS: From July 2000 to June 2004, 1988 patients with colorectal cancer were analysed and 114 CRC patients (5.7%) from 48 families were categorized as having HNPCC, including 76 from 26 families diagnosed clinically and 38 from the other 22 families diagnosed genetically. The sensitivity and specificity of high MSI and IHC for predicting mutations were 100% and 54%, and 79% and 77%, respectively. CONCLUSION: The frequency of HNPCC is approximately 10% among all Chinese CRC cases. The MSI and IHC detections for hMSH2/hMLH1 proteins are reliable pre-screening tests for hMLH1/hMSH2 germline mutations in families suspected of having HNPCC. PMID- 17461459 TI - Recurrence of autoimmune hepatitis after liver transplantation without elevation of alanine aminotransferase. AB - It is controversial whether steroid therapy should be continued to prevent the recurrence of autoimmune hepatitis (AIH) in patients who have undergone liver transplantation (LTx) due to AIH. We report a case of recurrent autoimmune hepatitis after LTx despite a persistently normal range of alanine aminotransferase (ALT). A 50-year-old woman was admitted to our hospital because of jaundice and severe liver dysfunction, where she was diagnosed with liver failure due to AIH. Steroid therapy was not effective enough and the patient received living-donor LTx in 1999. Following the operation, the level of ALT was maintained within a normal range and anti-nuclear antibody (ANA) became negative, however, the serum level of IgG gradually elevated and ANA became positive, while platelets decreased. A liver biopsy performed 6 years after LTx showed histological findings of AIH and she was diagnosed with recurrent AIH. A recurrence of AIH may occur after LTx even if the level of ALT remains within a normal range. We consider that a protocol liver biopsy should be performed in patients who undergo LTx due to AIH to decide the indication for steroid therapy. PMID- 17461460 TI - Intraductal papillary mucinous carcinoma with atypical manifestations: report of two cases. AB - Intraductal papillary mucinous neoplasms (IPMNs) are a well-characterized group of mucin-producing cystic neoplasms of the clear malignant potential type. We report here two cases of intraductal papillary mucinous carcinoma (IPMC) with atypical manifestations. In one case, we discussed a pseudomyxoma peritonei caused by a ruptured IPMC. In the other case we discussed the fistulization of IPMC into the stomach and duodenum. These two cases suggest that IPMN can either spontaneously rupture causing mucinous materials to spill into the free abdominal cavity or directly invade adjacent organs resulting in fistula development. PMID- 17461461 TI - Splenic abscess in a patient with fecal peritonitis. AB - Splenic abscess is a rare entity normally associated with underlying diseases. We report a case of splenic abscess with large gas formation in a non-diabetic and non-immunosuppressed patient after surgery for colon perforation. The most frequent cause of splenic abscess is septic embolism arising from bacterial endocarditis. Splenic abscess has a high rate of mortality when it is diagnosed late. Computed tomography resolved any diagnostic doubt, and subsequent surgery confirmed the diagnosis. PMID- 17461462 TI - Gynura root induces hepatic veno-occlusive disease: a case report and review of the literature. AB - Gynura root has been used extensively in Chinese folk medicine and plays a role in promoting microcirculation and relieving pain. However, its hepatic toxicity should not be neglected. Recently, we admitted a 62-year old female who developed hepatic veno-occlusive disease (HVOD) after ingestion of Gynura root. Only a few articles on HVOD induced by Gynura root have been reported in the literature. It is suspected that pyrrolizidine alkaloids in Gynura root might be responsible for HVOD. In this paper, we report a case of HVOD and review the literature. PMID- 17461463 TI - A rare case of pregnancy complicated by mesenteric mass: what does chylous ascites tell us? AB - Mesenteric fibromatosis is a rare benign nonmetastatic neoplasm that appears as a sporadic lesion or occurs in patients with familial polyposis, while chylous ascites associated with aggressive mesenteric fibromatosis during pregnancy has never been reported thus far. Here we present the case of a 28-year old pregnant woman, in whom an aggressive mesenteric fibromatosis with chylous ascites was detected, involving the jejunum, superior mesenteric artery (SMA) and superior mesenteric vein (SMV) and pancreas. One year after a successful surgical excision, the patient had no signs of recurrence. The authors report the case for its rarity and emphasize on combining clinicopathological, radiological and immunohistochemistry analysis for management of the disease. PMID- 17461464 TI - New methods for the management of esophageal varices. AB - Bleeding from esophageal varices (EVs) is a catastrophic complication of chronic liver disease. Many years ago, surgical procedures such as esophageal transection or distal splenorenal shunting were the only treatments for EVs. In the 1970s, interventional radiology procedures such as transportal obliteration, left gastric artery embolization, and partial splenic artery embolization were introduced, improving the survival of patients with bleeding EVs. In the 1980s, endoscopic treatment, endoscopic injection sclerotherapy (EIS), and endoscopic variceal ligation (EVL), further contributed to improved survival. We combined IVR with endoscopic treatment or EIS with EVL. Most patients with EVs treated endoscopically required follow-up treatment for recurrent varices. Proper management of recurrent EVs can significantly improve patients' quality of life. Recently, we have performed EVL at 2-mo (bi-monthly) intervals for the management of EVs. Longer intervals between treatment sessions resulted in a higher rate of total eradication and lower rates of recurrence and additional treatment. PMID- 17461465 TI - Role of computed tomographic colonoscopy of postoperative surveillance in patient with gastric cancer. AB - AIM: To examine the diagnostic yield of colorectal neoplasia at computed tomographic colonoscopy (CTC) as well as the feasibility of contrast enhanced CTC in patients with gastric cancer. METHODS: To examine the incidence of colon polyp we selected postoperative 188 gastric cancer patients, which we refer to as the 'colon polyp survey group'. To examine the feasibility of CTC for early detection of colon cancer or advanced colon adenoma, we selected 47 gastric cancer patients (M:F 29:18, mean age 53.8 years), which we call the 'CT colonoscopy group'. All the 47 patients underwent successive CTC and colonoscopy on the same day. RESULTS: Totally 109 colon polyps were observed from 59 out of 188 gastric cancer patients, the incidence rate of colon polyps in gastric cancer patients being 31.4%. The sensitivity of CTC in detecting individuals with at least 1 lesion of any size was 57.1%, the specificity was 72.7%, the positive predictive value was 47.1%, and the negative predictive value was 71.9%. When the cutoff size was decreased to 6 mm, the sensitivity and specificity were 80.0% and 92.9%, respectively, with positive and negative predictive values of 57.1% and 97.5%, respectively. Only one patient was classified as false negative by virtual colonoscopy. CONCLUSION: The diagnostic yield of colorectal polyp was 31.4% in patients with gastric cancer, and contrast enhanced CTC is an acceptable tool for the detection of synchronous colorectal advanced adenoma and postoperative surveillance of gastric cancer patients. PMID- 17461466 TI - Mechanism of apoptotic effects induced selectively by ursodeoxycholic acid on human hepatoma cell lines. AB - AIM: To investigate the effects of ursodeoxycholic acid (UDCA) on apoptosis and proliferation of hepatoma cell lines. METHODS: Human hepatoma cell lines HepG2 and BEL 7402 were cultured in medium supplemented with different concentrations of UDCA, normal human hepatic line L-02 was used as control. Cell proliferation, apoptosis and gene expression were detected using methyl thiazolyl tetrazolium (MTT) assay, flow cytometry, Western blot, DNA ladder assay, electron microscopy, and immunocytochemistry. RESULTS: Ursodeoxycholic acid inhibited the proliferation of HepG2 and BEL7402 cell lines in a dose-dependent manner. Ursodeoxycholic acid can change cell cycle distribution of HepG2 and BEL7402, the proportion of cells in G0-G1 phase increased whereas the proportion of S phase cells and G2-M phase cells decreased. Ursodeoxycholic acid arrested the cell cycle in G0-G1 phase by down-regulating the cell cycle related proteins cyclin D1, D3 and retinoblastoma protein (p(Rb)). The apoptotic rates of HepG2 and BEL7402 treated with UDCA (1.0 mmol/L) were significantly higher than those of control. In the HepG2 and BEL7402 treated with UDCA, expression of bcl-2 decreased whereas expression of Bax increased, the nuclear fragmentation and chromosomal condensed, cells shrank and lost attachment, apoptotic bodies and DNA ladders appeared. UDCA had no effect in inducing apoptosis on L-02 cell lines. CONCLUSION: UDCA can selectively inhibit proliferation and induce apoptosis of HepG2 and BEL7402 cell lines by blocking cell cycle and regulating the expression of Bax/bcl-2 genes. PMID- 17461467 TI - Sonic hedgehog-Gli1 pathway in colorectal adenocarcinomas. AB - AIM: To determine the role of Sonic hedgehog (Shh) pathway in colorectal adenocarcinomas through analysis of the expression of Shh pathway-related molecules, Shh, Ptch1, hedgehog-interacting protein (Hip), Gli1, Gli3 and PDGFRalpha. METHODS: Expression of Shh in 25 colorectal adeno-carcinomas was detected by RT-PCR, in situ hybridization and immunohistochemistry. Expression of Ptch1 was observed by in situ hybridization and immunohistochemistry. Expression of Hip, Gli1, Gli3 and PDGFRalpha was analyzed by in situ hybridization. RESULTS: Expression of cytokeratin AE1/AE3 was observed in the cytoplasm of colorectal crypts. Members of the Hh signaling pathway were expressed in colorectal epithelium. Shh was expressed in cytoplasm of dysplastic epithelial cells, while expression of Ptch1, Hip and Gli1 were mainly detected in the malignant crypts of adenocarcinomas. In contrast, PDGFRalpha was expressed highly in aberrant crypts and moderately in the stroma. Expression of Gli3 could not be detected in colorectal adenocarcinomas. CONCLUSION: These data suggest that Shh-Ptch1-Gli1 signaling pathway may play a role in the progression of colorectal tumor. PMID- 17461468 TI - Evaluation of the effect of pyrrolidine dithiocarbamate in suppressing inflammation in mice with dextran sodium sulfate-induced colitis. AB - AIM: To evaluate the effect of pyrrolidine dithio-carbamate (PDTC; an NF-kappaB inhibitor) administered at low (50 mg/kg) and high (100 mg/kg) doses in suppressing colitis in mice with dextran sodium sulfate (DSS)-induced colitis. METHODS: Mice were divided into a DSS-untreated group (normal group), DSS-treated control group, DSS+PDTC-treated group I (low-dose group), and DSS+PDTC-treated group II (high-dose group). In each group, the disease activity index score (DAI score), intestinal length, histological score, and the levels of activated NF kappaB and inflammatory cytokines (IL-1beta and TNF-alpha) in tissue were measured. RESULTS: The DSS+PDTC-treated group II exhibited suppression of shortening of intestinal length and reduction of DAI score. Activated NF-kappaB level and IL-1beta and TNF-alpha levels were significantly lower in DSS+PDTC treated group II. CONCLUSION: These findings suggest that PDTC is useful for the treatment of ulcerative colitis. PMID- 17461469 TI - Enhancement of insulin-producing cell differentiation from embryonic stem cells using pax4-nucleofection method. AB - AIM: To enhance the differentiation of insulin producing cell (IPC) ability from embryonic stem (ES) cells in vitro. METHODS: Four-day embryoid body (EB) formatted ES cells were dissociated as single cells for the followed plasmid DNA delivery. The use of Nucleofector electroporator (Amaxa biosystems, Germany) in combination with medium-contained G418 provided a high efficiency of gene delivery for advanced selection. Neucleofected cells were plated on the top of fibronectin-coated Petri dishes. Addition of Ly294002 and raised the glucose in medium at 24 h before examination. The differentiation status of these cells was monitored by semi-quantitative PCR (SQ-PCR) detection of the expression of relative genes, such as oct-4, sox-17, foxa2, mixl1, pdx-1, insulin 1, glucagons and somatostatin. The percentage of IPC population on d 18 of the experiment was investigated by immunohistochemistry (IHC), and the content/secretion of insulin was estimated by ELISA assay. The mice with severe combined immunodeficiency disease (SCID) pretreated with streptozotocin (STZ) were used to eliminate plasma glucose restoration after pax4+ ES implantation. RESULTS: A high efficiency of gene delivery was demonstrated when neucleofection was used in the present study; approximately 70% cells showed DsRed expression 2 d after neucleofection. By selection of medium-contained G418, the percentage of DsRed expressing cells kept high till the end of study. The pancreatic differentiation seemed to be accelerated by pax4 nucleofection. When compared to the group of cells with mock control, foxa2, mixl1, pdx1, higher insulin and somatostatin levels were detected by SQ-PCR 4 d after nucleofection in the group of pax4 expressing plasmid delivery. Approximately 55% of neucleofected cells showed insulin expression 18 d after neucleofection, and only 18% of cells showed insulin expression in mock control. The disturbance was shown by nucleofected pax4 RNAi vector; only 8% of cells expressed insulin 18 d after nucleofection. A higher IPC population was also detected in the insulin content by ELISA assay, and the glucose dependency was demonstrated in insulin secretion level. In the animal model, improvement of average plasma glucose concentration was observed in the group of pax-4 expressed ES of SCID mice pretreated with STZ, but no significant difference was observed in the group of STZ-pretreated SCID mice who were transplanted ES with mock plasmid. CONCLUSION: Enhancement of IPC differentiation from EB-dissociated ES cells can be revealed by simply using pax4 expressing plasmid delivery. Not only more IPCs but also pancreatic differentiation-related genes can be detected by SQ PCR. Expression of relative genes, such as foxa 2, mixl 1, pdx-1, insulin 1 and somatostatin after nucleofection, suggests that pax4 accelerates the whole differentiation progress. The higher insulin production with glucose dependent modulation suggests that pax4 expression can drive more mature IPCs. Although further determination of the entire mechanism is required, the potential of pax-4 nucleofected cells in medical treatment is promising. PMID- 17461470 TI - Upregulation of hypoxia inducible factor 1alpha mRNA is associated with elevated vascular endothelial growth factor expression and excessive angiogenesis and predicts a poor prognosis in gastric carcinoma. AB - AIM: To investigate the implication of the hypoxia inducible factor HIF-1alpha mRNA in gastric carcinoma and its relation to the expression of vascular endothelial growth factor (VEGF) protein, tumor angiogenesis invasion/metastasis and the patient's survival. METHODS: In situ hybridization was used to examine expression of HIF-1alpha mRNA, and immunohistochemical staining was used to examine expression of VEGF protein and CD34 in 118 specimens from patients with gastric carcinoma. RESULTS: The positive rates of HIF-1alpha mRNA and VEGF protein were 49.15% and 55.92%, respectively. Positive expressions of HIF-1alpha and VEGF in stage T3-T4 tumors and those with vessel invasion, lymph node metastasis and distant metastasis were dramatically stronger than stage T1-T2 cases and those without vessel invasion, lymph node metastasis and distant metastasis. The mean microvascular density (MVD) in stage T3-T4 tumors and those with vessel invasion, lymph node metastasis and distant metastasis was significantly higher than stage T1-T2 tumors and those without vessel invasion, lymph node metastasis and distant metastasis. The mean MVD in tumors with positive HIF-1alpha and VEGF expression was significantly higher than that in tumors with negative HIF-1alpha and VEGF expression. The expression of HIF-1alpha was positively correlated with VEGF protein. There were positive correlations between MVD and expression of HIF-1alpha and VEGF. The mean survival time and the 5-year survival rate in cases with positive expression HIF-1alpha and VEGF and MVD value >or= 41.5/0.72 mm2 were significantly lower than those with negative expression of HIF-1alpha and VEGF and MVD value < 41.5/0.72 mm2. CONCLUSION: Overexpression of HIF-1alpha is found in gastric carcinoma. HIF-1alpha may induce the angiogenesis in gastric carcinoma by upregulating the transcription of VEGF gene, and take part in tumor invasion and metastasis. They can be used as prognostic markers of gastric cancer in clinical practice. PMID- 17461471 TI - Aquaporin-8 expression is reduced in ileum and induced in colon of patients with ulcerative colitis. AB - AIM: To study susceptibility genes which may play a potential role in the pathogenesis and etiology of inflammatory bowel disease (IBD). METHODS: To identify potential susceptibility genes we performed global gene expression profiling in patients with IBD and control specimens. For determination of an intrinsic gene expression profile in ulcerative colitis (UC) and Crohn's disease (CD) compared to normal subjects, mucosal biopsies of non-inflamed regions of the colon and the terminal ileum were subjected to DNA microarray analysis. Real-time RT-PCR and immunohistochemistry were used for verification of selected regulated candidate genes and a genetic analysis was performed. RESULTS: We could show that aquaporin-8 (AQP8) mRNA and protein levels were significantly increased in the colon of UC patients compared to controls. Genetic analysis of the six exons and the promoter region of AQP8, however, revealed no mutations or polymorphisms in IBD patients. CONCLUSION: Our results suggest that upregulation of AQP8 in the colon of UC patients represents a secondary phenomenon which may, due to altered water exchange of the distal intestinal mucosa, disturb the physiologic colonic mucus barrier and thus lead to chronic inflammation and ulceration. PMID- 17461472 TI - Abdominal computed tomography in refractory coeliac disease and enteropathy associated T-cell lymphoma. AB - AIM: To evaluate computed tomography (CT) findings, useful to suggest the presence of refractory celiac disease (RCD) and enteropathy associated T cell lymphoma (EATL). METHODS: Coeliac disease (CD) patients were divided into two groups. Group I: uncomplicated CD (n = 14) and RCD type I (n = 10). Group II: RCD type II (n = 15) and EATL (n = 7). RESULTS: Both groups showed classic signs of CD on CT. Intussusception was seen in 1 patient in group I vs 5 in group II (P = 0.06). Lymphadenopathy was seen in 5 patients in group II vs no patients in group I (P = 0.01). Increased number of small mesenteric vessels was noted in 20 patients in group I vs 11 in group II (P = 0.02). Eleven patients (50%) in group II had a splenic volume < 122 cm3 vs 4 in group I (14%), 10 patients in group I had a splenic volume > 196 cm3 (66.7%) vs 5 in group II (33.3%) P = 0.028. CONCLUSION: CT scan is a useful tool in discriminating between CD and (Pre) EATL. RCD II and EATL showed more bowel wall thickening, lymphadenopathy and intussusception, less increase in number of small mesenteric vessels and a smaller splenic volume compared with CD and RCD I. PMID- 17461473 TI - Clinical significance of main pancreatic duct dilation on computed tomography: single and double duct dilation. AB - AIM: To study the patients with main pancreatic duct dilation on computed tomography (CT) and thereby to provide the predictive criteria to identify patients at high risk of significant diseases, such as pancreatic cancer, and to avoid unnecessary work up for patients at low risk of such diseases. METHODS: Patients with dilation of the main pancreatic duct on CT at Emory University Hospital in 2002 were identified by computer search. Clinical course and ultimate diagnosis were obtained in all the identified patients by abstraction of their computer database records. RESULTS: Seventy-seven patients were identified in this study. Chronic pancreatitis and pancreatic cancer were the most common causes of the main pancreatic duct dilation on CT. Although the majority of patients with isolated dilation of the main pancreatic duct (single duct dilation) had chronic pancreatitis, one-third of patients with single duct dilation but without chronic pancreatitis had pancreatic malignancies, whereas most of patients with concomitant biliary duct dilation (double duct dilation) had pancreatic cancer. CONCLUSION: Patients with pancreatic double duct dilation need extensive work up and careful follow-up since a majority of these patients are ultimately diagnosed with pancreatic cancer. Patients with single duct dilation, especially such patients without any evidence of chronic pancreatitis, also need careful follow-up since the possibility of pancreatic malignancy, including adenocarcinoma and intraductal papillary mucinous tumors, is still high. PMID- 17461474 TI - Effect of omeprazole and domperidone on adult asthmatics with gastroesophageal reflux. AB - AIM: To study the effect of combined omeprazole (Ome) and domperidone (Dom) therapy on asthma symptoms and pulmonary function in asthmatics with gastroesophageal reflux. METHODS: We selected 198 asthmatics with gastroesophageal reflux diagnosed by 24-h esophageal pH monitoring to receive Ome 20 mg twice daily and Dom 10 mg three times daily or placebo for 16 wk (1:1 double-blind randomization). Spirometry was done at baseline and after 16 wk of treatment. The primary outcome measures were: mean daily daytime and nighttime asthma symptom scores. Mean daily reflux symptom scores, albuterol use as rescue medication (number of puffs), daytime and nighttime peak expiratory flow rate (PEFR), postbronchodilator forced expiratory volume in 1 second (FEV1) and postbronchodilator forced vital capacity (FVC) were secondary outcome measures. RESULTS: Comparison of mean change from baseline between antireflux therapy and placebo groups revealed significant reduction in daytime asthma symptom score (17.4% vs 8.9%), nighttime asthma symptom score (19.6% vs 5.4%), reflux symptom score (8.7% vs 1.6%) and rescue medication use (23.2% vs 3.1%) after antireflux therapy compared to mean change in placebo group (P < 0.001). There was significant improvement in morning PEFR (7.9% vs 0.2%), evening PEFR (9.8% vs 0.5%), FEV1 (11.1% vs 3.78%) and FVC (9.3% vs 1.52%) in the antireflux therapy group compared to placebo on comparing the mean change from baseline after 16 wk (P < 0.01). CONCLUSION: Combined therapy with Ome and Dom in adult asthmatics with gastroesophageal reflux may be beneficial by reducing asthma symptoms, rescuing medication use, and improving pulmonary function. PMID- 17461475 TI - Diagnostic criteria for acute liver failure due to Wilson disease. AB - AIM: To describe the diagnostic criteria for acute liver failure due to Wilson disease (WD), which is an uncommon cause of acute liver failure (ALF). METHODS: We compared findings of patients presenting with ALF due to WD to those with ALF of other etiologies. RESULTS: Previously described criteria, such as low alkaline phosphatase activity, ratio of low alkaline phosphatase to total bilirubin or ratio of high aspartate aminotransferase (AST) to alanine aminotransferase (ALT), failed to identify patients with ALF due to WD. There were significant differences in low ALT and AST activities (53 +/- 43 vs 1982 +/- 938, P < 0.0001 and 87 +/- 44 vs 2756 +/- 2941, P = 0.037, respectively), low choline esterase activity (1.79 +/- 1.2 vs 4.30 +/- 1.2, P = 0.009), high urine copper concentrations (93.4 +/- 144.0 vs 3.5 +/- 1.8, P = 0.001) and low hemoglobin (7.0 +/- 2.2 vs 12.6 +/- 1.8, P < 0.0001) in patients with ALF caused by WD as compared with other etiologies. Interestingly, 4 of 7 patients with ALF due to WD survived without liver transplantation. CONCLUSION: In ALF, these criteria can help establish a diagnosis of WD. Where applicable, slit-lamp examination for presence of Kayser-Fleischer rings and liver biopsy for determination of hepatic copper concentration still remain important for the diagnosis of ALF due to WD. The need for liver transplantation should be evaluated carefully as the prognosis is not necessarily fatal. PMID- 17461476 TI - Coeliac disease in Dutch patients with Hashimoto's thyroiditis and vice versa. AB - AIM: To define the association between Hashimoto's thyroiditis and coeliac disease in Dutch patients. METHODS: A total of 104 consecutive patients with Hashimoto's thyroiditis underwent coeliac serological tests (antigliadins, transglutaminase and endomysium antibodies) and HLA-DQ typing. Small intestinal biopsy was performed when any of coeliac serological tests was positive. On the other hand, 184 patients with coeliac disease were subjected to thyroid biochemical (thyroid stimulating hormone and free thyroxine) and thyroid serological tests (thyroglobulin and thyroid peroxidase antibodies). RESULTS: Of 104 patients with Hashimoto's thyroiditis, sixteen (15%) were positive for coeliac serology and five patients with documented villous atrophy were diagnosed with coeliac disease (4.8%; 95% CI 0.7-8.9). HLA-DQ2 (and/or -DQ8) was present in all the five and 53 patients with Hashimoto's thyroiditis (50%; 95% CI 43-62). Of 184 patients with coeliac disease, 39 (21%) were positive for thyroid serology. Based on thyroid biochemistry, the 39 patients were subclassified into euthyroidism in ten (5%; 95% CI 2-9), subclinical hypothyroidism in seven (3.8%; 95% CI 1.8-7.6), and overt hypothyroidism (Hashimoto's thyroiditis) in 22 (12%; 95% CI 8-16). Moreover, four patients with coeliac disease had Graves' disease (2%; 95% CI 0.8-5) and one patient had post-partum thyroiditis. CONCLUSION: The data from a Dutch population confirm the association between Hashimoto's thyroiditis and coeliac disease. Screening patients with Hashimoto's thyroiditis for coeliac disease and vice versa is recommended. PMID- 17461477 TI - Ileoscopy in patients with ileocolonic tuberculosis. AB - AIM: To study the role of retrograde terminal ileoscopy in patients suspected to have ileocolonic tuberculosis. METHODS: A retrospective analysis was performed for patients undergoing colonoscopy for suspected ileocolonic tuberculosis between January 2000 and June 2004, in whom retrograde ileoscopy had been performed. Only patients diagnosed with tuberculosis on the basis of histological findings of either a caseating granuloma or those having non-caseating granuloma or a collection of epithelioid cells at a minimum of one endoscopic lesion (either in the colon or the terminal ileum) on histology with good responses to conventional anti-tuberculous drugs were enrolled for the study. RESULTS: Fifty three patients were included. The terminal ileum was involved in only 11 patients. Eight of these patients had involvement of the cecum too. Two patients had ileal lesions without cecal involvement; however, lesions were noted in the ascending colon. In one patient the whole colon was normal and only the terminal ileum showed nodularity and ulceration. Histological examination of the ileal biopsies obtained from the ileal lesions showed noncaseating granulomas in five, collection of epithelioid cells in four and nonspecific histology in two patients. Mucosal biopsies obtained from the lesion in the terminal ileum, in the patient in whom mucosal abnormality was observed to be confined to the ileum, showed noncaseating granulomas. In two other patients the ileal biopsies alone showed histological evidence of tuberculosis with biopsies from the colonic lesions showing non-specific inflammatory changes only. CONCLUSION: Retrograde ileoscopy should be performed in all patients undergoing colonoscopy for suspected ileocolonic tuberculosis. In some patients only the terminal ileum may be involved and histological examination may reveal evidence of tuberculosis only from the lesions in the terminal ileum. This approach would lead to additional lesions being picked up and increasing the chances of well-timed diagnosis of tuberculosis. PMID- 17461478 TI - pRB expression in esophageal mucosa of individuals at high risk for squamous cell carcinoma of the esophagus. AB - AIM: To investigate the pRb expression in a large group of patients with history of chronic exposure to the main risk factors for development of squamous cell carcinoma of the esophagus. METHODS: One hundred and seventy asymptomatic individuals at high risk for esophageal squamous cell carcinoma (consumption of more than 80 g of ethanol and 10 cigarettes/d for at least 10 years) underwent upper gastrointestinal endoscopy with biopsies of the esophageal mucosa. As a control group, specimens of esophageal mucosa obtained from 20 healthy subjects were also studied. Immunohistochemical assessment of the tissues was performed using a monoclonal antibody anti-pRB protein. RESULTS: Absence of the pRB staining, indicating loss of RB function, was observed in 33 (19.4%) of the individuals at risk for esophageal cancer, but in none of the healthy controls (P < 0.02). Loss of pRb expression increased in a stepwise fashion according to the severity of the histological findings (P < 0.005): normal mucosa (11/97 or 11.3%), chronic esophagitis (17/60 or 28.3%), low-grade dysplasia (3/10 or 30%), high-grade dysplasia 1/2 or 50%) and squamous cell carcinoma (1/1 or 100%). CONCLUSION: Our findings suggest that abnormal expression of the pRB protein may be implicated in the process of esophageal carcinogenesis. Additional studies are warranted to define the role of the pRB protein as a biomarker for development of esophageal squamous cell carcinoma in individuals at high risk for this malignancy. PMID- 17461479 TI - Increased intestinal permeability in pathogenesis and progress of nonalcoholic steatohepatitis in rats. AB - AIM: To investigate whether increased intestinal permeability contributes to the pathogenesis and progress of nonalcoholic steatohepatitis by observing its dynamic change in rat models. METHODS: Rat models of nonalcoholic steatohepatitis were established by giving a fat-rich diet. The rats were sacrificed at wk 8, 12 and 16 during the study. Rats fed with normal diet were taken as control. Plasma D-lactate, plasma diamine oxidase, serum lipids and liver transaminases were measured in blood of the femoral artery. Hepatic steatosis and inflammation were assessed by haematoxylin-eosin staining. RESULTS: A rat model of nonalcoholic steatohepatitis was established successfully. Plasma D-lactate level in model group at wk 8, 12 and 16 and diamine oxidase level in model group at wk 12, 16 increased significantly compared with those in control group. There were notable differences of D-lactate and diamine oxidase level in model group between wk 8 and 12 as well as between wk 12 and 16. Serum lipids, liver transaminases and liver injury also increased with disease development. CONCLUSION: Increased intestinal permeability caused by intestinal bacterial overgrowth and endotoxin induced intestinal destruction exists in rats with nonalcoholic steatohepatitis, which may partially explain the pathogenesis and progress of this disease. PMID- 17461480 TI - Construction and analysis of tree models for chromosomal classification of diffuse large B-cell lymphomas. AB - AIM: To construct tree models for classification of diffuse large B-cell lymphomas (DLBCL) by chromosome copy numbers, to compare them with cDNA microarray classification, and to explore models of multi-gene, multi-step and multi-pathway processes of DLBCL tumorigenesis. METHODS: Maximum-weight branching and distance-based models were constructed based on the comparative genomic hybridization (CGH) data of 123 DLBCL samples using the established methods and software of Desper et al. A maximum likelihood tree model was also used to analyze the data. By comparing with the results reported in literature, values of tree models in the classification of DLBCL were elucidated. RESULTS: Both the branching and the distance-based trees classified DLBCL into three groups. We combined the classification methods of the two models and classified DLBCL into three categories according to their characteristics. The first group was marked by +Xq, +Xp, -17p and +13q; the second group by +3q, +18q and +18p; and the third group was marked by -6q and +6p. This chromosomal classification was consistent with cDNA classification. It indicated that -6q and +3q were two main events in the tumorigenesis of lymphoma. CONCLUSION: Tree models of lymphoma established from CGH data can be used in the classification of DLBCL. These models can suggest multi-gene, multi-step and multi-pathway processes of tumorigenesis. Two pathways, -6q preceding +6q and +3q preceding +18q, may be important in understanding tumorigenesis of DLBCL. The pathway, -6q preceding +6q, may have a close relationship with the tumorigenesis of non-GCB DLBCL. PMID- 17461481 TI - Comparison of biological characteristics of marrow mesenchymal stem cells in hepatitis B patients and normal adults. AB - AIM: To establish a culture system of marrow mesenchymal stem cells (MSCs) from hepatitis B patients and normal adults and to compare their biological characteristics. METHODS: MSCs were isolated from bone marrow in 34 male hepatitis B patients and 15 male normal adults and cultivated in vitro. Their biological characteristics including surface markers, shapes and appearances, growth curves, first passage time and passage gene-rations were compared. RESULTS: Cultivation achievement ratio of hepatitis B patients was lower than that of normal adults, no statistical significance (82.35% vs 100%, P > 0.05). Compared with MSCs of normal adults, MSCs of hepatitis B patients presented a statistical lower growth curve, longer first passage time (13.0 +/- 1.6 d vs 11.4 +/- 1.5 d, P < 0.05), fewer passaging generation numbers (10.5 +/- 1.4 generations vs 12.3 +/- 1.7 generations, P < 0.05), though both shared same appearances, shapes and surface markers. MSCs in hepatitis B patients would expand, spread out and age more easily and there were more refractive particles in the cytoplasm. CONCLUSION: MSCs from hepatitis B patients can be cultured in vitro. Although their appearance, shape and surface marker are similar to those of MSCs from normal adults, there are differences in their biological characteristics. PMID- 17461482 TI - Expression and significance of intratumoral interleukin-12 and interleukin-18 in human gastric carcinoma. AB - AIM: To explore the effect of intratumoral expressions of interleukin-12 (IL-12) and interleukin-18 (IL-18) on clinical features, angiogenesis and prognosis of gastric carcinoma. METHODS: The expressions of IL-12 and IL-18 from 50 samples of gastric cancer tissue were analyzed by immunohistochemistry, and microvessel density (MVD) was determined with microscopic imaging analysis system. RESULTS: The positive expression rates of IL-12 and IL-18 were 44% (22/50) and 26% (13/50), respectively. IL-12 was significantly associated with pathologic differentiation, depth of invasion, lymph node metastasis, distant metastasis, and TNM stage, and IL-18 was closely related to distant metastasis. Intratumoral IL-12 and IL-18 expressions were not statistically related to MVD scoring. IL-12 positive patients survived significantly longer than those with IL-12-negative tumors, but there was no significant difference between IL-18-positive patients and IL-18-negative ones. The multivariate analysis with Cox proportional hazard model revealed IL-12, MVD and T stage were independent prognostic factors. CONCLUSION: The positive expressions of IL-12 and IL-18 can play an important role in progression and metastasis of gastric cancer, and IL-12 might be an independent factor of poor prognosis in gastric carcinoma. PMID- 17461483 TI - Enlarged accessory spleen presenting stomach submucosal tumor. AB - A 62-year-old man presented with upper abdominal discomfort underwent upper gastrointestinal endoscopy. Gastroscopy and endoscopic ultrasonography revealed a submucosal tumor (SMT) with homogenous echogenicity originated from extragastric organs. An abdominal contrast-enhanced computed tomography (CT) showed that the well-marginated ovoid mass, approximately 6 cm in diameter, enhanced homogenously to a similar degree as splenic parenchyma. (99m)Technetium sulfur colloid scintigraphy revealed the splenic nature of the mass. A diagnosis of accessory spleen mimicking gastric SMT was made. Subsequent follow-up was uneventful without performing splenectomy. PMID- 17461484 TI - Common carotid arterial thrombosis associated with ulcerative colitis. AB - A 26-year-old woman with ulcerative colitis was transferred to our hospital with left hemiparesis due to cerebral infarction. Cervical ultrasonography and magnetic resonance imaging angiography revealed thrombosis at the right common carotid artery and the right internal carotid artery. Antithrombotic and anticoagulant therapies were commenced. After about 2 wk of the treatment, the frequency of her diarrhea increased. She underwent emergency subtotal colectomy, but 10 d later an abundant hemorrhage from the remnant rectum occurred, so the remnant rectum was resected and an ileal pouch anal anastomosis was performed. Antithrombotic and anticoagulant therapies were continued, but neither her neurological status nor magnetic resonance imaging angiography findings showed subsequent changes. She was discharged 3 mon after operation. This is a rare case of common carotid arterial thrombosis occurring as a complication of ulcerative colitis, in which antithrombotic and anticoagulant therapies are considered to provoke a deterioration of the patient's bowel disease. PMID- 17461485 TI - Eosinophilic gastroenteritis presenting as small bowel obstruction: a case report and review of the literature. AB - Eosinophilic gastroenteritis is a rare disease of unknown etiology. It is characterized by eosinophilic infiltration of the bowel wall to a variable depth and symptoms associated with gastrointestinal tract. Recently, the authors experienced a case of eosinophilic gastroenteritis presenting as small bowel obstruction. A 51-year old woman was admitted to our hospital complaining of abdominal pain and vomiting. Physical examination revealed a distended abdomen with diffuse tenderness. Complete blood count showed mild leukocytosis without eosinophilia. Computed tomography confirmed a dilatation of the small intestine with ascites. An emergency laparotomy was performed for a diagnosis of peritonitis due to intestinal obstruction. Segmental resection of the ileum and end to end anastomosis were performed. Histologically, there was a dense infiltration of eosinophils throughout the entire thickness of ileal wall and eosinophilic enteritis was diagnosed. The patient recovered well, and was free from gastrointestinal symptoms at the time when we reported her disease. PMID- 17461486 TI - Bitter gourd (Momordica charantia): a potential mechanism in anti-carcinogenesis of colon. PMID- 17461487 TI - Innovative therapeutics for inflammatory bowel disease. AB - Inflammatory bowel diseases (IBD) are chronic inflammatory conditions of the gastrointestinal tract, which clinically present as one of two disorders, Crohn's disease or ulcerative colitis. Mainstays of drug treatments for IBD include aminosalicylates, corticosteroids and immunosuppressants such as azathioprine, methotrexate and cyclosporin. Advances in basic research of the pathophysiological process in IBD have been applied to generate a variety of new therapeutics targeting at different levels of the inflammatory processes. New therapies are classified as: (1) Anti-TNFalpha antibodies; (2) Recombinant cytokines; (3) Selective adhesion blockade; (4) Growth factors; (5) Innate immunostimulation; (6) Nucleic acid based therapies; (7) Gene therapy; (8) Autologous bone-marrow transplantation; (9) Helminths and (10) Extracorporeal immunomodulation. All treatments have the potential to provide more effective and safe treatment for IBD. PMID- 17461489 TI - Clinical characteristics of idiopathic portal hypertension. AB - Idiopathic portal hypertension is one of the interesting causes of portal hypertension. Even in very developed medical centers, this disorder is still one of the most important misdiagnoses of clinical practice. To inexperienced physicians, presenting esophageal varices and upper gastrointestinal bleeding usually prompt an unfortunate diagnosis of cirrhosis. A heterogenous clinical presentation and progression of this disorder should be recognized by physicians, and management should be directed towards some specific problems confined to this disorder. Although a genetic basis and other factors are implicated in its pathogenesis, exact underlying mechanism(s) is (are) unknown. In this review, we discuss the heterogeneity of idiopathic portal hypertension, its etiopathogenesis, clinical presentation and management issues. With the expectation of an excellent prognosis, a practicing gastroenterologist should be aware that "not all varices mean cirrhosis". PMID- 17461488 TI - Treatment of hepatitis C virus infection. AB - Acute and chronic hepatitis C virus (HCV) infection remains a serious health problem worldwide, however, there has been advancement in the treatment of HCV infection due to standard treatment using pegylated interferon and ribavirin. The literature indicates that therapy for HCV is becoming more individualized. In addition to considering genotype and viral RNA levels before treatment, achievement of an early virologic response (EVR) and a rapid virologic response (RVR) is now possible during therapy. Moreover, problem patients, such as non responders, relapsers, HIV or HBV co-infected patients, patients with liver cirrhosis, and pre- or post-liver transplantation patients are an increasing fraction of the patients requiring treatment. This article reviews the literature regarding standard treatments and problem patients with acute and chronic HCV infection. It also includes discussion on contraindications and side effects of treatment with interferon and ribavirin, as well as new drug development. PMID- 17461491 TI - Is portal vein cavernous transformation a component of congenital hepatic fibrosis? AB - Congenital hepatic fibrosis (CHF) is an autosomal recessive disorder that belongs to the family of fibropolycystic liver diseases. This family includes a spectrum of disorders which are usually found in combination with each other and are usually inherited. Clinically fibropolycystic diseases have three effects being present in different proportions, those of a space occupying lesion, of portal hypertension and of cholangitis. In most patients, the first manifestations of CHF are signs and symptoms related to portal hypertension such as splenomegaly and varices. Portal hypertension in these patients has been attributed to the hypoplasia or compression of the portal vein radicles in the fibrous bands. Cavernous transformation of the portal vein (CTPV) is a relatively rare condition resulting from extrahepatic portal vein obstruction with recanalization or collateral vein formation to bypass the obstruction. It has been found that patients with CHF having an accompanying CTPV have relatively large splenomegaly and suffers more frequent episodes of bleeding from esophageal varices. We believe that CTPV is a congenital component of CHF and also one of the important causative factors of portal hypertension in these patients. PMID- 17461492 TI - Clinical characteristics of Caroli's disease. AB - Caroli's disease is a rare congenital condition chara-cterized by non-obstructive saccular or fusiform dilatation of larger intrahepatic bile ducts. Cholangitis, liver cirrhosis, and cholangiocarcinoma are its potential complications. The diagnosis of Caroli's disease depends on demonstrating that the cystic lesions are in continuity with the biliary tree which can be showed by ultrasonography, computerized tomography, endoscopic retrograde cholangiopancreatography, percutaneous transhepatic cholangiography or magnetic resonance cholangiopancreatography. Treatment of Caroli's disease relies on the location of the biliary abnormalities. While localized forms confined to one lobe can be treated with surgery, liver transplantation is the only effective modality for diffuse forms. Although a rare disorder; Caroli's disease should always be considered in the differential diagnosis of chronic cholestasis of unknown cause. PMID- 17461490 TI - Hepatic venous outflow obstruction: three similar syndromes. AB - Our goal is to provide a detailed review of veno-occlusive disease (VOD), Budd Chiari syndrome (BCS), and congestive hepatopathy (CH), all of which results in hepatic venous outflow obstruction. This is the first article in which all three syndromes have been reviewed, enabling the reader to compare the characteristics of these disorders. The histological findings in VOD, BCS, and CH are almost identical: sinusoidal congestion and cell necrosis mostly in perivenular areas of hepatic acini which eventually leads to bridging fibrosis between adjacent central veins. Tender hepatomegaly with jaundice and ascites is common to all three conditions. However, the clinical presentation depends mostly on the extent and rapidity of the outflow obstruction. Although the etiology and treatment are completely different in VOD, BCS, and CH; the similarities in clinical manifestations and liver histology may suggest a common mechanism of hepatic injury and adaptation in response to increased sinusoidal pressure. PMID- 17461493 TI - Clinical characteristics of Caroli's syndrome. AB - Caroli's syndrome is characterized by multiple segmental cystic or saccular dilatations of intrahepatic bile ducts associated with congenital hepatic fibrosis. The clinical features of this syndrome reflect both the characteristics of congenital hepatic fibrosis such as portal hypertension and that of Caroli's disease named as recurrent cholangitis and cholelithiasis. The diagnosis depends on both histology and imaging methods which can show the communication between the sacculi and the bile ducts. Treatment consists of symptomatic treatment of cholangitis attacks by antibiotics, some endoscopic, radiological and surgical drainage procedures and surgery. Liver transplantation seems the ultimate treatment for this disease. Prognosis is fairly good unless recurrent cholangitis and renal failure develops. PMID- 17461494 TI - Suppression of human colon tumor growth by adenoviral vector-mediated NK4 expression in an athymic mouse model. AB - AIM: To investigate the suppressive effects of adenoviral vector-mediated expression of NK4, an antagonist of hepatocyte growth factor (HGF), on human colon cancer in an athymic mouse model to explore the possibility of applying NK4 to cancer gene therapy. METHODS: A human colon tumor model was developed by subcutaneous implantation of tumor tissue formed by LS174T cells grown in athymic mice. Fifteen tumor-bearing mice were randomized into three groups (n = 5 in each group) at d 3 after tumor implantation and mice were injected intratumorally with phosphate-buffered saline (PBS) or with recombinant adenovirus expressing beta galactosidase (Ad-LacZ) or NK4 (rvAdCMV/NK4) at a 6-d interval for total 5 injections in each mouse. Tumor sizes were measured during treatment to draw a tumor growth curve. At d 26 after the first treatment, all animals were sacrificed and the tumors were removed to immunohistochemically examine proliferating cell nuclear antigen (PCNA), microvessel density (represented by CD31), and apoptotic cells. In a separate experiment, 15 additional athymic mice were employed to develop a tumor metastasis model by intraperitoneal injection (ip) of LS174T cells. These mice were randomized into 3 groups (n = 5 in each group) at d 1 after injection and were treated by ip injection of PBS, or Ad LacZ, or rvAdCMV/NK4 at a 6-d interval for total two injections in each mouse. All animals were sacrificed at d 14 and the numbers and weights of disseminated tumors within the abdominal cavity were measured. RESULTS: Growth of human colon tumors were significantly suppressed in the athymic mice treated with rvAdCMV/NK4 (2537.4 +/- 892.3 mm(3)) compared to those treated by either PBS (5175.2 +/- 1228.6 mm(3)) or Ad-LacZ (5578.8 +/- 1955.7 mm(3)) (P < 0.05). The tumor growth inhibition rate was as high as 51%. Immunohistochemical staining revealed a similar PCNA labeling index (75.1% +/- 11.2% in PBS group vs 72.8% +/- 7.6% in Ad LacZ group vs 69.3% +/- 9.4% in rvAdCMV/NK4 group) in all groups, but significantly lower microvessel density (10.7 +/- 2.4 in rvAdCMV/NK4 group vs 25.6 +/- 3.8 in PBS group or 21.3 +/- 3.5 in Ad-LacZ group, P < 0.05), and a markedly higher apoptotic index (7.3% +/- 2.4% in rvAdCMV/NK4 group vs 2.6 +/- 1.1% in PBS group or 2.1% +/- 1.5% in Ad-LacZ group, P < 0.05) in the rvAdCMV/NK4 group compared to the two control groups. In the tumor metastasis model, the number and weight of disseminated tumors of mice treated with rvAdCMV/NK4 were much lower than those of the control groups (tumor number: 6.2 +/- 3.3 in rvAdCMV/NK4 group vs 22.9 +/- 7.6 in PBS group or 19.8 +/- 8.5 in Ad-LacZ group, P < 0.05; tumor weight: 324 +/- 176 mg in rvAdCMV/NK4 group vs 962 +/- 382 mg in PBS group or 1116 +/- 484 mg in Ad-LacZ group, P < 0.05). CONCLUSION: The recombinant adenovirus, rvAdCMV/NK4, can attenuate the growth of colon cancer in vivo, probably by suppressing angiogenesis and inducing tumor cell apoptosis, but not by direct suppression of tumor cell proliferation. Moreover, rvAdCMV/NK4 may inhibit peritoneal dissemination of colon cancer cells in a murine tumor metastasis model. These findings indicate that NK4 gene transfer may be an effective tool for the treatment of colon cancer. PMID- 17461495 TI - Celecoxib attenuates 5-fluorouracil-induced apoptosis in HCT-15 and HT-29 human colon cancer cells. AB - AIM: To investigate the combined chemotherapeutic effects of celecoxib when used with 5-FU in vitro. METHODS: Two human colon cancer cell lines (HCT-15 and HT-29) were treated with 5-FU and celecoxib, alone and in combination. The effects of each drug were evaluated using the MTT [3- (4, 5-dimethylthiazol-2-yl)-2, 5 diphenyltetrazolium bromide] assay, flow cytometry, and western blotting. RESULTS: 5-FU and celecoxib showed a dose-dependent cytotoxic effect. When treated with 10(-3) mol/L 5-FU (IC(50)) and celecoxib with its concentration ranging from 10(-8) mol/L to 10(-4) mol/L of celecoxib, cells showed reduced cytotoxic effect than 5-FU (10(-3) mol/L) alone. Flow cytometry showed that celecoxib attenuated 5-FU induced accumulation of cells at subG1 phase. Western blot analyses for caspase-3 and poly (ADP-ribose) polymerase (PARP) cleavage showed that celecoxib attenuated 5-FU induced apoptosis. Western blot analyses for cell cycle molecules showed that G2/M arrest might be possible cause of 5-FU induced apoptosis and celecoxib attenuated 5-FU induced apoptosis via blocking of cell cycle progression to the G2/M phase, causing an accumulation of cells at the G1/S phase. CONCLUSION: We found that celecoxib attenuated cytotoxic effect of 5 FU. Celecoxib might act via inhibition of cell cycle progression, thus preventing apoptosis induced by 5-FU. PMID- 17461496 TI - Protective effect of curcumin against liver warm ischemia/reperfusion injury in rat model is associated with regulation of heat shock protein and antioxidant enzymes. AB - AIM: To investigate the hypothesis that the protective effects of curcumin in hepatic warm ischemia/reperfusion (I/R) injury are associated with increasing heat shock protein 70 (Hsp70) expression and antioxidant enzyme activity. METHODS: Sixty Sprague-Dawley male rats were randomly divided into sham, I/R, C + I/R groups. The model of reduced-size liver warm ischemia and reperfusion was used. Curcumin (50 mg/kg) was administered by injection through a branch of superior mesenteric vein at 30 min before ischemia in C + I/R group. Five rats were used to investigate the survival during 1 wk after operation in each group. Blood samples and liver tissues were obtained in the remaining animals after 3, 12, and 24 h of reperfusion to assess serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), liver tissue NO(2)(-) + NO(3)(-), malondialdehyde (MDA) content, superoxide dismutase (SOD), catalase (CAT), nitricoxide synthase (NOS) and myeloperoxidase (MPO) activity, Hsp70 expression and apoptosis ratio. RESULTS: Compared with I/R group, curcumin pretreatment group showed less ischemia/reperfusion-induced injury. CAT and SOD activity and Hsp70 expression increased significantly. A higher rate of apoptosis was observed in I/R group than in C + I/R group, and a significant increase of MDA, NO(2)(-) + NO(3)(-) and MPO level in liver tissues and serum transaminase concentration was also observed in I/R group compared to C + I/R group. Curcumin also decreased the activity of inducible NO synthase (iNOS) in liver after reperfusion, but had no effect on the level of endothelial NO synthase (eNOS) after reperfusion in liver. The 7 d survival rate was significantly higher in C + I/R group than in I/R group. CONCLUSION: Curcumin has protective effects against hepatic I/R injury. Its mechanism might be related to the overexpression of Hsp70 and antioxidant enzymes. PMID- 17461497 TI - Lactobacillus plantarum inhibits epithelial barrier dysfunction and interleukin-8 secretion induced by tumor necrosis factor-alpha. AB - AIM: To determine whether Lactobacillus plantarum can modify the deleterious effects of tumor necrosis factor-alpha (TNF-alpha) on intestinal epithelial cells. METHODS: Caco-2 cells were incubated with TNF-alpha alone or in the presence of L. plantarum. Transepithelial electrical resistance was used to measure epithelial barrier function. Interleukin 8 (IL-8) secretion by intestinal epithelial cells was measured using an ELISA. Cellular lysate proteins were immunoblotted using the anti-extracellular regulated kinase (ERK), anti-phospho ERK and anti-IkappaB-alpha. RESULTS: A TNF-alpha-induced decrease in transepithelial electrical resistance was inhibited by L. plantarum. TNF-alpha induced IL-8 secretion was reduced by L. plantarum. L. plantarum inhibited the activation of ERK and the degradation of IkappaB-alpha in TNF-alpha-treated Caco 2 cells. CONCLUSION: Induction of epithelial barrier dysfunction and IL-8 secretion by TNF-alpha is inhibited by L. plantarum. Probiotics may preserve epithelial barrier function and inhibit the inflammatory response by altering the signal transduction pathway. PMID- 17461498 TI - Timing of mortality in severe acute pancreatitis: experience from 643 patients. AB - AIM: To determine the timing of mortality after onset of severe acute pancreatitis (SAP) and the course of the disease in a large series of patients. METHODS: From July 1996 to June 2005, all patients diagnosed with acute pancreatitis at Chang Gung Memorial Hospital, Taipei, Taiwan were retrospectively studied. Three thousand two hundred and fifty episodes of acute pancreatitis were recorded in 2248 patients (1431 males and 817 females; median age, 55.6 years; range, 18-97 years). Mortality was divided into two groups: early death (<= 14 d after admission), and late death (> 14 d after admission). The clinical features of patients in these two groups were compared. RESULTS: Although the overall mortality rate of acute pancreatitis was 3.8% (123/3250), mortality rate of SAP was as high as 16.3% (105/643). Of those 105 SAP mortalities, 44 (41.9%) deaths occurred within the first 14 d after admission and 61 (58.1%) occurred after14 d. Incidence of early death did not significantly differ from that of late death. The co-morbidities did not contribute to the timing of death. Early deaths mainly resulted from multiple organ failure. Late deaths were mainly caused by secondary complication of infected necrosis. Intra-abdominal bleeding significantly caused higher mortality in late death. CONCLUSION: Approximately half (42%) of SAP deaths occur within 14 d and most were due to multiple organ failure. The late deaths of SAP were mostly due to infected necrosis. PMID- 17461499 TI - Perioperative management of primary liver cancer. AB - AIM: To minimize the complications and mortality and improve the survival in primary liver cancer (PLC) patients undergoing hepatic resection. METHODS: We conducted a retrospective analysis of 2143 PLC patients treated from January 1990 to January 2004. The patients were divided into two groups using January 1997 as a cut-off. Small tumor size (< 5 cm), preoperative redox tolerance index (RTI), vascular control method, and postoperative arterial ketone body ratio (AKBR) were used as indicators of surgical outcome. RESULTS: Small tumors had less complications and lower mortality and higher overall survival rate. Use of RTI for selecting patients and types of hepatectomy, reduced complications (21.1% vs 11.0%) and mortality (1.6% vs 0.3%). The half liver vascular occlusion protocol (n = 523) versus the Pringle method (n = 476) showed that the former significantly reduced the postoperative complications (25.8% vs 11.9%) and mortality (2.3% vs 0.6%) respectively, and cut mean hospital stay was 3.5 d. Postoperative AKBR was a reliable indicator of the energy status in survivors. CONCLUSION: RTI is of value in predicting hepatic functional reserve, half liver occlusion could protect the residual liver function, and AKBR measurement is a simple and accurate means of assessing the state of postoperative metabolism. Optimal perioperative management is an important factor for minimizing complications and mortality in patients undergoing hepatic resection. PMID- 17461500 TI - Detection of hepatitis B virus genotypes using oligonucleotide chip among hepatitis B virus carriers in Eastern China. AB - AIM: To determine the genotype distribution of hepatitis B virus (HBV) with a newly oligonucleotide chip assay among the HBV carriers in Eastern China. METHODS: An assay using oligonucleotide chip was developed for detection of HBV genotypes in serum samples from HBV DNA-positive patients in Eastern China. This method is based on the principle of reverse hybridization with Cy5-labeled amplicons hybridizing to type-specific oligonucleotide probes that are immobilized on slides. The results of 80 randomly chosen sera were confirmed by direct sequencing. RESULTS: HBV genotype B, C and mixed genotype were detected in 400 serum samples, accounting for 8.3% (n = 33), 83.2% (n = 333), and 8.5% (n = 34), respectively. The evaluation of the oligonucleotide assay showed 100% concordance with the amplicon phylogenetic analysis except 9 mixed genotype infections undetected by sequencing. CONCLUSION: The study indicates that HBV genotype C and B prevail in the Eastern China. It is suggested that the oligonucleotide chip is a reliable and convenient tool for the detection of HBV genotyping. PMID- 17461501 TI - Recto-vestibular disruption defect resulted from the malpractice in the treatment of the acquired recto-vestibular fistula in infants. AB - AIM: To explore the pathogenesis of the rectovestibular disruption (RVD) defect and to recommend a successful repair, and prevention of it. METHODS: Clinical records of 15 girls, age ranged from 3 to 15 (median, 7.5) years, with acquired rectovestibular fistula (RVF) mistreated before were retrospectively reviewed. All of them presented an abnormal appearance of perineum and were suffering from some degree of fecal incontinence, and those were graded III to IV by Li Zheng's Score. Repair of anal sphincters and reconstruction of perineum body and skin by anterior perineal rectoanoplasty were performed in all cases. RESULTS: Operation in all cases was successful. The perineum looked practically normal and fecal continence score rose up to VI by Li Zheng's Score. CONCLUSION: The conventional treatment for anal fistula, lay-open or string-treatment, should be considered as malpractice of RVF, and certainly leads to the RVD defect, and the anterior perineal rectoanoplasty could cure it satisfactorily. PMID- 17461502 TI - Pioglitazone attenuates the severity of sodium taurocholate-induced severe acute pancreatitis. AB - AIM: To determine the effect of pioglitazone, a specific peroxisome proliferator activated receptor-gamma (PPARgamma) ligand, on development of severe acute pancreatitis (SAP) and expression of nuclear factor-kappa B (NF-kappaB) and intercellular adhesion molecule-1 (ICAM-1) in the pancreas. METHODS: Male Sprague Dawley (SD) rats (160-200 g) were randomly allocated into three groups (n = 18 in each group): severe acute pancreatitis group, pioglitazone group, sham group. SAP was induced by retrograde infusion of 1 mL/kg body weight 5% sodium taurocholate (STC) into the biliopancreatic duct of male SD rats. Pioglitazone was injected intraperitoneally two hours piror to STC infusion. Blood and ascites were obtained for detecting amylase and ascitic capacity. Pancreatic wet/dry weight ratio, expression of NF-kappaB and ICAM-1 in pancreatic tissues were detected by immunohistochemical staining. Pancreatic tissue samples were stained with hematoxylin and eosin (HE) for routine optic microscopy. RESULTS: Sham group displayed normal pancreatic structure. SAP group showed diffuse hemorrhage, necrosis and severe edema in focal areas of pancreas. There was obvious adipo saponification in abdominal cavity. Characteristics such as pancreatic hemorrhage, necrosis, severe edema and adipo-saponification were found in pioglitazone group, but the levels of those injuries were lower in pioglitazone group than those in SAP group. The wet/dry pancreatic weight ratio, ascetic capacity, serum and ascitic activities of anylase in the SAP group were significantly higher than those in the sham group and pioglitazone group respectively (6969.50 +/- 1368.99 vs 2104.67 +/- 377.16, 3.99 +/- 1.22 vs 2.48 +/ 0.74, P < 0.01 or P < 0.05). According to Kusske criteria, the pancreatic histologic score showed that interstitial edema, inflammatory infiltration, parenchyma necrosis and parenchyma hommorrhage in SAP group significantly differed from those in the sham group and pioglitazone group (7.17 +/- 1.83 vs 0.50 +/- 0.55, 7.67 +/- 0.82 vs 6.83 +/- 0.75, P < 0.01, P < 0.05. The expression of NF-kappaB and ICAM-1 in sham group was lower than that in SAP group and pioglitazone group (0.50 +/- 0.55 vs 33 +/- 1.21, P < 0.01). There was a significant difference in the expression of NF-kappaB and ICAM-1 between SAP group and pioglitazone group (7.50 +/- 1.05 vs 11.33 +/- 1.75, 0.80 +/- 0.53 vs 1.36 +/- 0.54, P < 0.01 or P < 0.05) at 12 h after the induction of pancreatitis. CONCLUSION: Pioglitazone attenuates the severity of SAP. The beneficial effect of pioglitazone is multifactorial due to its anti-inflammatory activities, most likely through the inhibition of ICAM-1 expression and NF-kappaB activation. Specific ligands of PPARgamma may represent the novel and effective means of clinical therapy for SAP. PMID- 17461503 TI - Enhanced therapeutic effects of combined chemotherapeutic drugs and midkine antisense oligonucleotides for hepatocellular carcinoma. AB - AIM: To evaluate the effect of combined antisense oligonucleotides targeting midkine (MK-AS) and chemotherapeutic drugs [cisplatin(DDP), 5-fluorouracil (5-FU) and adriamycin (ADM)] on inhibition of HepG2 cell proliferation, and to analyze the efficacy of MK-AS used in combined ADM in in situ human hepatocellular carcinoma (HCC) model. METHODS: HepG2 cells were treated with MK-AS and/or chemotherapeutic drugs mediated by Lipofectin, and cell growth activity was determined by MTS assay. An in situ HCC model was used in this experiment. MK-AS, ADM and MK-AS + ADM were given intravenously for 20 d, respectively. The animal body weight and their tumor weight were measured to assess the effect of the combined therapy in vivo. RESULTS: Combined treatment with MK-AS reduced the IC50 of DDP, 5-FU and ADM in HepG2 cells. MK-AS significantly increased the inhibition rate of DDP, 5-FU and ADM. Additionally, synergism (Q 1.15) occurred at a lower concentration of ADM, 5-FU and DDP with combined MK-AS. Combined treatment with MK-AS and ADM resulted in the more growth inhibition on in situ human HCC model compared with treatment with chemotherapeutic drugs alone. CONCLUSION: MK-AS increases the chemosensitivity in HepG2 cells and in situ human HCC model, and the combination of MK-AS and ADM has a much better in vitro and in vivo synergism. PMID- 17461504 TI - A case of eosinophilic cholangitis: imaging findings of contrast-enhanced ultrasonography, cholangioscopy, and intraductal ultrasonography. AB - A 38-year-old woman was referred to our institution due to epigastralgia. She presented with obstructive jaundice and eosinophilia. Endoscopic retrograde cholangiopancreatography showed diffuse narrowing from the distal common bile duct to the bifurcation of the hepatic ducts. An endoscopic plastic biliary stent was inserted; the specimen obtained from the common bile duct wall revealed dense infiltration by eosinophils. Treatment was started with prednisolone 60 mg daily. The patient's biliary stenosis and eosinophilia gradually improved. Eosinophilic infiltration in the lungs or stomach is relatively common, but it is rare in the common bile duct. Most of the reported cases of eosinophilic cholangitis presented with eosinophilia; our patient's eosinophil count was over 1000/mm(3). Since our patient had allergies to pollen and house dust, a relationship between the allergies and the eosinophilic cholangitis was suspected, but no cause was identified. PMID- 17461505 TI - An unhappy triad: hemochromatosis, porphyria cutanea tarda and hepatocellular carcinoma-a case report. AB - Liver fibrosis and cirrhosis are predisposing factors for the development of hepatocellular carcinoma (HCC). Hemosiderosis has also been described to trigger carcinogenesis. A significant iron overload, as found in hereditary hemochromatosis (HHC), is a risk factor for HCC and may also promote the symptoms of porphyria cutanea tarda (PCT). A 68-year old male patient presented to our clinic with a suspected HCC, elevated alpha-fetoprotein but normal liver function tests. He reported a 25 year-old history of vitiligo upon exposure to sunlight. The patient underwent an extended left hemihepatectomy, and the recovery was uneventful, with the exception of a persistent hyperbilirubinemia. Perfusion problems and extrahepatic cholestasis were ruled out by CT-scan with angiography and MR-cholangiopancreatography. However, MRI showed an iron overload. Histology confirmed the HCC (pT3, pN0, G3, R0) and revealed a portal fibrosis and hemosiderosis. Based on the skin lesions we suspected a PCT that was confirmed by laboratory tests showing elevated porphyrin, uroporphyrin, coproporphyrin and porphobilinogen. Concurrently, molecular diagnostics revealed homozygosity for the C282Y mutation within the hemochromatosis HFE gene. After phlebotomy and normalization of liver function tests the patient was discharged. This is the first case ever showing the unusual combination of HCC in a fibrotic liver with HHC and PCT. This diagnosis not only warrants oncological follow-up but also symptomatic therapy to normalize iron metabolism and thereby improve liver function and alleviate the symptoms of HHC and PCT. Thus progression of fibrosis may be prevented and liver regeneration supported. PMID- 17461506 TI - Treatment of pediatric Ogilvie's syndrome with low-dose erythromycin: a case report. AB - Acute colonic pseudo-obstruction is a poorly understood syndrome, characterized by the signs, symptoms and radiological pattern of a large bowel obstruction without evidence for a mechanical obstruction. We report a case of a 2-year old boy who presented with progressive abdominal distention, vomiting and abdominal pain on postoperative d 3. Plain abdominal z-ray showed markedly dilated large bowel. Mechanical colonic obstruction was ruled out with hypaque enema. Ogilvie's syndrome was suspected. The patient received treatment with oral erythromycin which had an immediate beneficial effect. During the 6 mo follow-up, no recurrences of symptoms were observed. We provide a safe and effective therapy for Ogilvie's syndrome in pediatric individuals. PMID- 17461507 TI - An incidental case of triple gallbladder. AB - Triplication of the gallbladder is a very rare congenital anomaly of the biliary tract; there are only eleven reported cases to date. Gallbladder multiplications are not likely to be discovered unless associated with cholelithiasis, sludge, cholecystitis and carcinoma. Here we report an incidentally diagnosed triplicate gallbladder in a patient with sigmoid diverticulitis; two of the triplicate gallbladder were demonstrated with ultrasound and computed tomography, and an additional galballder was found at surgery. PMID- 17461508 TI - Pure red cell aplasia due to parvovirus B19 infection after liver transplantation: a case report and review of the literature. AB - Pure red cell aplasia (PRCA) due to parvovirus B19 (PVB19) infection after solid organ transplantation has been rarely reported and most of the cases were renal transplant recipients. Few have been described after liver transplantation. Moreover, little information on the management of this easily recurring disease is available at present. We describe the first case of a Chinese liver transplant recipient with PVB19-induced PRCA during immunosuppressive therapy. The patient suffered from progressive anemia with the lowest hemoglobin level of 21 g/L. Bone marrow biopsy showed selectively inhibited erythropoiesis with giant pronormoblasts. Detection of PVB19-DNA in serum with quantitative polymerase chain reaction (PCR) revealed a high level of viral load. After 2 courses of intravenous immunoglobulin (IVIG) therapy, bone marrow erythropoiesis recovered with his hemoglobin level increased to 123 g/L. He had a low-level PVB19 load for a 5-mo follow-up period without recurrence of PRCA, and finally the virus was cleared. Our case indicates that clearance of PVB19 by IVIG in transplant recipients might be delayed after recovery of anemia. PMID- 17461509 TI - Development of multiple myeloma in a patient with gastrointestinal stromal tumor treated with imatinib mesylate: a case report. AB - Gastrointestinal stromal tumors (GISTs) are rare tumors, which represent approximately 1% of the neoplasms of the gastrointestinal tract. These tumors rarely give extra-abdominal metastases. However, their clinical outcome is potentially adverse. In some rare cases, co-existance of GISTs with other malignancies has been reported. Here we present a case of a 74-year old male with GIST, which was managed by surgical resection. Fourteen months later, the patient presented with liver metastases and imatinib mesylated was administered. During treatment, the patient reported skeletal pain and plane X-rays revealed osteolytic bone lesions. Further investigation revealed the presence of multiple myeloma. To the best of our knowledge, this is the first report of the co existence of multiple myeloma (MM) with GIST. PMID- 17461511 TI - Meta-analysis: some clinical and statistical contributions in several medical disciplines. AB - Meta-analysis in its present form of statistically integrating information from several studies all with a common underlying theme has been around for over 25 years. The medical field has seen many attempts by many investigators to pull summary data together from various sources within a discipline with the goal of making some definitive statement about the state of the science in that discipline. Likewise authors of manuscripts in the background and rationale section of their paper always summarize what they believe to be the state of affairs up to the time of the presentation of their own results in that particular paper. The new data and results they present in their current publication is an attempt to update the progress in that field. Thus in a sense they have performed a partial meta- analysis of summarizing information from the past, presenting their added contribution and thus updating the knowledge base. They have not quite integrated past data in a rigorous statistical way with their new data, but have merely used the data history to justify their current research which pretty much stands on its own. Thus meta-analysis is an after the fact attempt to pull together the current knowledge base whether it be publications or raw data and present a statistical synthesis of all the information and reach a conclusion as to the best treatment or intervention strategy based on all these past contributions. Now it's time to look back at some of these meta-analyses and determine what contributions, if any, they have made to the knowledge base within certain medical disciplines. Many disciplines including psychiatry have been visited by meta- analysis. One now examines some of these studies in the areas of oncology, orthopedics, psychiatry, pediatrics and cardiology. The purpose is to determine, given the information presented, what contributions, statistical challenges and peripheral issues in these disciplines have been brought to light in these meta- analyses. PMID- 17461510 TI - Use of grafts in pelvic reconstructive surgery. AB - There has been growing interest in the use of grafts in pelvic reconstructive surgery. This article will address available graft materials and assess their clinical efficacy and safety. We conducted a Pubmed MEDLINE literature search for full-length English text studies with follow-up periods of at least one year. There are many reports on synthetic and biological graft materials; the majority are not well-designed, have short-term follow-up, small sample sizes, and poor outcome assessment. The use of non-absorbable synthetic grafts may offer excellent anatomical cure rates. However, it is associated with a high incidence of graft-related complications, including healing abnormalities and adverse bladder, bowel, and sexual function effects. These complications can be decreased with absorbable synthetic meshes, but efficacy is lower compared to non absorbable ones. There is insufficient evidence in favor of biological grafts. In conclusion, based on current knowledge, routine application of grafts in pelvic reconstruction is not recommended. It is preferred that graft utilization be individualized, with close monitoring for complications. PMID- 17461512 TI - Risk factors influencing the occurrence of injuries in koreans requiring hospitalization. AB - PURPOSE: This study focused on local communities and the factors influencing injuries requiring hospital admission that affect the quality of life for Koreans. MATERIALS AND METHODS: This study used data collected from 4,400 households during the 2001 Korea National Health and Nutrition Examination Survey (KNHANES). A stratified multistage probability sampling method was applied and the final sample included 7,924 subjects over 20 years old who had completed the questionnaire regarding factors influencing injuries requiring hospital admission. Multiple logistic regression analysis was used to estimate the risk factors influencing injuries. RESULTS: The incidence of injuries requiring hospital admission in Koreans was higher in men than in women at 2.3% (95% CI: 1.8-2.8) and 1.7% (95% CI: 1.3-2.1), respectively. Statistically significant factors affecting the occurrence of injuries requiring hospital admission were marital status (OR: 2.60, 95% CI: 2.22- 5.56), state of health (OR: 0.63, 95% CI: 0.43-0.92), frequency of heavy drinking (OR: 1.96, 95% CI: 1.01-3.79), normal physical activities (OR: 1.68, 95% CI: 1.07-2.65), and sleep (OR: 1.54, 95% CI: 1.01-2.35). CONCLUSION: The results of this study can be used to develop measures to prevent fatal injuries and be used as basic data for community health programs. PMID- 17461513 TI - Clinical role of bone marrow angiogenesis in childhood acute lymphocytic leukemia. AB - PURPOSE: Vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) are associated with increased angiogenesis, growth, and metastasis in solid tumors. But, until today, the importance of theses factors on leukemia, especially childhood acute lymphocytic leukemia (ALL) has received limited attention. Therefore, this study examined the bone marrow plasma VEGF and bFGF levels in ALL patients and normal controls. PATIENTS AND METHODS: Bone marrow plasmas at diagnosis from 33 ALL patients (median age 5.9 years; range 1.8-13.9 years) were used for analysis. The bone marrow levels of bFGF and VEGF were determined by enzyme-linked immunosorbent assay (R & D Systems) and compared with the bone marrow levels of 7 healthy control subjects (median age 11.98 years; 6 months -13.6 years). RESULTS: Average VEGF was higher in relapse ALL (N=7, 216.6 +/- 79.9pg/mL) compared to standard (N=9, 36.8 +/- 12.1pg/mL) (p=0.013) or high risk ALL (N=17, 80.0 +/- 12.2pg/mL) (p=0.023). bFGF levels were also significantly higher in relapse than standard-, or high-risk ALL patients (relapse ALL; 48.6 +/- 15.4pg/mL, standard risk ALL; 18.9 +/- 5.5pg/mL, high risk ALL; 19.0 +/- 3.5pg/mL, normal control; 18.6 +/- 4.0pg/mL) (p=0.003). Three patients with refractory relapse and death had much higher VEGF and bFGF values (VEGF; 420.0 +/- 81.6pg/ mL, bFGF; 85.6 +/- 3.2pg/mL). CONCLUSION: Our data suggest that the increased levels of VEGF and bFGF in bone marrow may play an important role in prognosis of childhood ALL. PMID- 17461514 TI - Accessory gene regulator group polymorphisms in methicillin-resistant Staphylococcus aureus: an association with clinical significance. AB - PURPOSE: Virulent gene expression in Staphylococcus aureus is controlled by regulators such as the accessory gene regulator (agr). Strains can be divided into four major agr groups (agr I-IV) on the basis of agrD and agrC polymorphisms. The purpose of this study was to define the proportion of agr I, II, and III polymorphisms and to compare the clinical characteristics between group I and non-group I polymorphisms of methicillin-resistant Staphylococcus aureus (MRSA) strains in a Korean tertiary care teaching hospital. MATERIALS AND METHODS: A total of 158 clinical isolates were evaluated by RFLPs (restriction fragment length polymorphisms). RESULTS: The mean age of the patients was 50.2 +/ 21.9 years old. There were 74 (49.3%), 66 (44.0%), 10 (6.7%), 7 (4.4%), and 1 (0.6%) strains in agr group I, II, III, I + II, and I + III polymorphisms, respectively. Only ear infections were a statistically significant clinical parameter according to univariate (p=0.001) and multivariate analysis (OR, 4.721 (1.273-17.508), p=0.020). CONCLUSION: This study suggests that agr group I is the most prevalent in Korea, and ear infections are correlated with the group I polymorphism, which is a different clinical trend from western countries. It can also be inferred that community-acquired MRSA correlates with agr group I. PMID- 17461515 TI - Hospital staff and patient recognition toward opening of medical services market, and factors in selecting a foreign hospital. AB - PURPOSE: The objectives of this study are to compare the hospital employees' and patients' recognition and attitudes toward the opening of the medical services market, to analyze the differences between hospital employees and patients on the factors in selecting a foreign hospital. MATERIALS AND METHODS: This study collected and analyzed data using systematic questionnaires that were self administered by employees and outpatients to compare their recognition of the opening of the medical services market and the factors involved in selecting a foreign hospital. RESULTS: Employees exhibited a higher level of recognition than the patients. Reasons for agreement were to ensure the right of selection for both the employees and patients. Reasons for disagreement, however, were split between the two groups: degraded public characteristics of medical services, for employees; and expensive medical fees, for patients. The most urgent task to prepare for the opening of the medical services market is the improvement of the diagnosis and treatment technologies for employees, and improvement of the services provided by hospital staff for patients. CONCLUSION: Korean hospitals shall need to improve their diagnosis and treatment technologies and provide medical services of high quality in order to compete with foreign hospitals. PMID- 17461516 TI - Associations between smoking, screening, and death caused by cervical cancer in Korean women. AB - PURPOSE: Cervical cancer is a major women's health problem in the world today. The purpose of this study was to estimate the incidence and mortality rates and to investigate risk factors for cervical cancer in Korean women. MATERIALS AND METHODS: Reproductive factors, cigarette smoking, as well as the risk of incidence and death from cervical cancer were examined in a 12-year prospective cohort study of 475,398 Korean women aged 30 to 95 years who received health insurance from the National Health Insurance Corporation and who had a medical evaluation in 1992. Relative risks (RR) and 95% confidence intervals (CI) were calculated using the Cox proportional hazards model after adjusting for age, body mass index, cigarette smoking, alcohol use, menarche, parity, and Papanicolaou test status. RESULTS: This study showed that the RR of death due to cervical cancer among current smokers was two times higher compared with non- smokers (RR=2.00; 95% CI, 1.23-2.91). In addition, the RR of death due to cervical cancer among all women who smoked > or = 10 cigarettes/day was 2.4 times higher than the RR among women that had never smoked. More interestingly, those who had never been screened by Papanicolaou smears had twice the risk of death due to cervical cancer (RR =2.00; 95% CI, 1.37-1.81). CONCLUSION: Our prospective study concluded that current smokers had an increased risk of death due to cervical cancer. We suggest that the target age group for cervical cancer screening tests be reconsidered and should begin as early as possible. PMID- 17461517 TI - Influence of combined methionine synthase (MTR 2756A > G) and methylenetetrahydrofolate reductase (MTHFR 677C > T) polymorphisms to plasma homocysteine levels in Korean patients with ischemic stroke. AB - PURPOSE: Methionine synthase (MTR) and 5,10-methylenetetrahydrofolate reductase (MTHFR) are the main regulatory enzymes for homocysteine metabolism. The present case- control study was conducted to determine whether there is an association between the MTR 2756A > G or MTHFR 677C > T polymorphism and plasma homocysteine concentration in Korean subjects with ischemic stroke. MATERIALS AND METHODS: DNA samples of 237 patients who had an ischemic stroke and 223 age and sex-matched controls were studied. MTR 2756A > G and MTHFR 677C > T genotypes were determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). RESULTS: Frequencies of mutant alleles for MTR and MTHFR polymorphisms were not significantly different between the controls and cases. The patient group, however, had significantly higher homocysteine concentrations of the MTR 2756AA and MTHFR 677TT genotypes than the control group (p=0.04 for MTR, p=0.01 for MTHFR). The combined MTR 2756AA and MTHFR 677TT genotype (p= 0.04) and the homocysteine concentrations of the patient group were also higher than those of the controls. In addition, the genotype distribution was significant in the MTHFR 677TT genotype (p=0.008) and combined MTR 2756AA and MTHFR 677TT genotype (p=0.03), which divided the groups into the top 20% and bottom 20% based on their homocysteine levels. CONCLUSION: The results of the present study demonstrate that the MTR 2756A > G and MTHFR 677C > T polymorphisms interact with elevated total homocysteine (tHcy) levels, leading to an increased risk of ischemic stroke. PMID- 17461518 TI - What are the causative factors for a slow, progressive enlargement of a chronic subdural hematoma? AB - PURPOSE: To test the hypothesis that chronic subdural hematoma (CSDH) enlarges by the causative factors, this study has performed. MATERIALS AND METHODS: In 10 patients with CSDH, coagulation factors in venous blood taken at the time of surgery and hematomic contents aspirated from the CSDH were studied, using both laboratory assays and microscopy. RESULTS: When compared to the range of normal plasma, the hematoma fluids demonstrated a marked reduction in factor II, V, VII, VIII, and X, moderate reduction of factors IX and XI, and slight reduction of factor XII. Activated protein C and antithrombin III levels were decreased. The FDP (Fibrinogen Degradation Product) levels in chronic subdural hematoma were extremely high. The endothelial cells of the macrocapillaries (also called ''sinusoid'') showed numerous gap junctions between adjacent endothelial cells and a thinness or absence of the basement membrane, suggesting that the macrocapillaries are very fragile and susceptible to bleeding. CONCLUSION: Excessive coagulation in the hematoma, predominantly via the extrinsic clotting pathway, local hyperfibrinolysis, transmitted pulsations, and characteristics of the macrocapillaries play an important role in the leakage of blood and the enlargement of CSDH. PMID- 17461519 TI - Serum amyloid a as a useful indicator of disease activity in patients with ankylosing spondylitis. AB - PURPOSE: To investigate whether serum amyloid A (SAA) levels are increased in patients with ankylosing spondylitis (AS) and whether its levels correlate well with AS disease activity. MATERIALS AND METHODS: Thirty-eight patients with AS and 38 age- and sex-matched control subjects were enrolled in this cross sectional study. Their SAA levels were quantitatively measured by immunonephelometry. An established, self-administered instrument for evaluating disease activity (Bath Ankylosing Spondylitis Disease Activity Index, BASDAI) was used to measure and acute phase reactants, including erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP), in patients with AS. RESULTS: Patients with AS had a significantly higher mean SAA level than controls (9.52 +/- 7.49mg/L versus 2.73 +/- 1.57mg/L, p < 0.05), and the mean BASDAI score of patients with elevated SAA levels was significantly higher than that of patients with normal SAA levels (5.6 +/- 1.3 versus 4.4 +/- 1.5, p < 0.05). SAA levels showed significant correlations with BASDAI scores (r=0.431, p=0.007), ESR (r=0.521, p=0.001) and CRP levels (r=0.648, p < 0.001). Additionally, the correlation between ESR and CRP levels also appeared significant (r=0.703, p < 0.001). In those with normal ESR or CRP levels, SAA levels and BASDAI scores were elevated (p < 0.05) and showed a trend of positive correlation with one another. CONCLUSION: Our data showed that SAA levels were increased in patients with AS and correlated well with disease activity. These findings suggest that SAA can be used as a valuable indicator of disease activity in AS. PMID- 17461520 TI - Polypoidal choroidal vasculopathy in Korean patients with large submacular hemorrhage. AB - PURPOSE: To determine and compare the clinical characteristics, visual prognosis and treatment of hemorrhagic polypoidal choroidal vasculopathy (HPCV) with those of hemorrhagic choroidal neovascularization (HCNV) due to age-related macular degeneration (ARMD). MATERIALS AND METHODS: Retrospective analysis of 44 consecutive eyes with a submacular hemorrhage comprising more than 50% of the neovascular lesion. Patients were diagnosed as having HPCV or HCNV on the basis of indocyanine green angiography. RESULTS: Of the 44 eyes with submacular hemorrhage, 26 were classified as HPCV and 18 as HCNV. The baseline patient characteristics were similar for both groups. At the final follow-up the HPCV group had 17 eyes showing visual improvement, four showing maintained vision, and five showing visual deterioration. In contrast, the HCNV group had four eyes showing visual improvement, one showing maintained vision, and 13 showing visual deterioration. Visual acuity of < 0.1 at follow-up was found in 7 (27%) of HPCV eyes and 10 (56%) of HCNV eyes. PDT was performed in 15 HPCV eyes, of which 13 (87%) showed improvement or no change in visual acuity, while only 2 (22%) of the 9 HCNV eyes responded similarly after PDT. Eyes treated with PDT did not have better outcomes compared to eyes that underwent other types of treatment (Fisher's exact test, p > 0.05). CONCLUSION: PCV accounts for the largest proportion of submacular hemorrhage in Koreans. PCV showed a better visual prognosis than CNV. PMID- 17461521 TI - Polymorphisms of CYP1A1 and GSTM1 genes and susceptibility to oral cancer. AB - PURPOSE: Oral cancer is the fifth most common form of cancer in the world and comprises 6.5% of all cancer deaths. Since one of the major risk factors for oral cancer is tobacco use, we hypothesized that polymorphic genes coding for tobacco carcinogen-metabolizing enzymes may play a role in oral cancer susceptibility. MATERIALS AND METHODS: To investigate the association between polymorphisms of the CYP1A1 and GSTM1 genes and risks for oral squamous cell carcinoma (OSCC) in the Korean population, the prevalence of the CYP1A1 Mspl and GSTM1 null polymorphisms were examined in 72 patients with histologically confirmed primary OSCC, as well as in 221 healthy control subjects. RESULTS: A significant risk increase for oral cancer was observed among subjects with the homozygous CYP1A1 (m2/m2) genotype (OR=3.8, 95% CI=1.9-7.7), but not the GSTM1 null genotype (OR=0.7, 95% CI=0.4-1.3). Risk for oral cancer was significantly increased in subjects with the homozygous CYP1A1 (m2/m2)genotype, regardless of smoking history (smokers; OR=4.4; 95% CI=1.2-16.3; non- smokers OR=4.9; 95% CI=1.9-12.5). Using the potentially most protective genotype GSTM1 (+)/CYP1A1 [(m1/m1)+ (m1/m2)] as the reference group, an increased risk for oral cancer was observed among subjects with the GSTM1 (+)/ CYP1A1 (m2/m2) (OR= 2.0, 95% CI=0.8-5.2), and GSTM1 (-)/ CYP1A1 (m2/m2) (OR=4.9, 95% CI=1.5-15.5) genotypes (p < 0.009, (chi2 trend test). CONCLUSION: Our results suggest that individuals with a genotype of CYP1A1 (m2/m2) and GSTM1 (-) are highly susceptible for OSCC and that the CYP1A1 (m2/m2) genotype is closely associated with increased risk of OSCC in Koreans. PMID- 17461522 TI - Nodular gastritis and pathologic findings in children and young adults with Helicobacter pylori infection. AB - PURPOSE: The aim of this study was to investigate the pathologic characteristics of nodular gastritis in children and young adults infected with Helicobacter pylori (H. pylori). MATERIALS AND METHODS: A total of 328 patients were enrolled in this study, and the diagnosis of H. pylori infection was done with gastroduodenal endoscopy concomitant with a CLO(TM) test and pathologic analysis of the biopsy specimens. Diagnoses of normal, superficial gastritis, nodular gastritis, and peptic ulcer disease were made from the gastroduodenal endoscopic findings. The density of H. pylori organisms in the gastric mucosa was rated as normal, mild, moderate, or marked. The pathologic findings of nodular gastritis were based on the histopathologic findings of inflammation, immune activity, glandular atrophy and intestinal metaplasia. Each of these findings was scored as either normal (0), mild (1), moderate (2), or marked (3) according to the updated Sydney system and using visual analog scales. The gastritis score was the sum of the four histopathologic scores. RESULTS: In this study, nodular gastritis (50.6%) was most common, and mild density (51.5%) H. pylori infection was also common upon microscopic examination. Intestinal metaplasia occurred in 9 patients (2.7%). CONCLUSION: Logistic regression revealed a significant increase in the incidence of nodular gastritis with gastritis score (p=0.008), but not an association with sex, age, or H. pylori density. Gastritis score was the only significant factor influencing the occurrence of nodular gastritis. Intestinal metaplasia, which was originally thought to be a pre-malignant lesion, occurred in 2.7% of the patients with H. pylori infection. PMID- 17461523 TI - Abduction motion analysis of hemiplegic shoulders with a fluoroscopic guide. AB - PURPOSE: We investigated the usefulness of video based, fluoroscopically guided abduction motion analysis of hemiplegic shoulders. PATIENTS AND METHODS: Twenty two stroke patients with Brunnstrom stages 3-4 (Group 1) or 5-6 (Group 2) were enrolled in this study. Patients with shoulder pain and significant spasticity (MAS 2) were excluded. We recorded motion pictures of the abductions of affected and unaffected shoulder joints under an AP fluoroscopic guide. Lateral scapular slide distances (D1: T2- superior angle, D2: T3- scapular spine, D3: T7-inferior angle) were measured at 30 degrees , 60 degrees , 90 degrees during glenohumeral abduction in a captured photographic image. The angles of scapular rotation and trajectory (stromotion) of the humeral head center, relative to the 3rd thoracic spine in the abduction motion were analyzed. RESULTS: In Group 1, a significant difference was found in the lateral scapular slide distance between the affected and sound sides. However, no significant side to side difference was found in Group 2. Scapular angles in abduction were also increased in Group 1. Patients with a more synergistic movement pattern showed less scapular stabilizing muscle activity and, instead, exhibited a compensatory ''shrugging'' like motion accomplished by spinal tilting. CONCLUSION: The present findings support the notion that the above parameters of fluoroscopically guided shoulder abduction motion analysis correlate well with clinical findings. These parameters should be useful for evaluations of hemiplegic shoulder biomechanics. PMID- 17461524 TI - Refractures of the upper extremity in children. AB - PURPOSE: To investigate the etiologic factors related to refractures of the upper extremity in children. PATIENTS AND METHODS: Eighteen refractures were divided into three groups according to the location of the initial fractures. They were analyzed in terms of the type of refractures, fracture patterns, and the existence of an underlying deformity. RESULTS: Of nine supracondylar fractures of the humerus, two involved refractures at the supracondylar region, and the other seven involved the lateral condyle. Underlying cubitus varus was present in six cases. Of three lateral condylar fractures of the humerus, one had a refracture at the supracondylar region, and two cases involved the lateral condyle. One had an underlying cubitus varus. All but one case in the humeral fractures group were late refractures, and were treated with surgery. Of six repeat forearm fractures, five were early type and occurred at the original site within nine weeks, four at the diaphysis of both bones of the forearm, and one at the diaphysis of the ulna. All cases in the forearm fractures group, save one, had volar angulation before the refracture, and were treated conservatively. CONCLUSION: In the humerus, the underlying cubitus varus was the most important predisposing factor for refractures and lateral condyle fractures were common. In the forearm, volar angulation of the diaphysis was related to refractures, and complete and circular consolidation of the primary fracture of the forearm was thought to be important in prevention. PMID- 17461525 TI - Thrombosuction utilizing an export aspiration catheter during primary percutaneous coronary intervention in acute myocardial infarction. AB - PURPOSE: Effective myocardial reperfusion after primary PCI for an AMI in lesions with a thrombus is limited by distal embolization and the slow/no reflow phenomenon. We evaluated the efficacy of a thrombus reduction technique using an export aspiration catheter for thrombosuction during primary PCI. MATERIALS AND METHODS: We analyzed 62 patients with AMIs who underwent primary PCI and had a thrombi burden during thrombosuction using an EAC (EAC group; n=31) or without thrombosuction (control group; n=31). RESULTS: Thrombosuction with an EAC was performed safely in all the patients in EAC group without any complications. After the PCI, restoration to a TIMI flow grade 3 was significantly more frequent in the EAC group (26/31 vs. 20/31, p < 0.05). However, the TIMI perfusion grade did not differ between the two groups. Further, the corrected TIMI frame counts were lower in the EAC group (23.9 +/- 15.1 vs. 34.8 +/- 22.5, p < 0.05). Although there was no statistical significance, a greater incidence of distal embolization was observed in the control group (16.1%, 5/31) as compared to the EAC group (0/31) (p= 0.056). However, the incidence of major adverse cardiac events at 1 and 6 months did not differ between the two groups. CONCLUSION: For AMIs, thrombosuction with an EAC before or during PCI is a safe and potentially effective method for restoration of the coronary flow. PMID- 17461526 TI - Percutaneous treatment of renal cysts with OK-432 sclerosis. AB - PURPOSE: The aim of this study was to demonstrate OK- 432 sclerotherapy efficacy for treatment of simple renal cysts. MATERIALS AND METHODS: Twenty patients with 25 symptomatic or large simple cysts were treated by ultrasonography (US)-guided percutaneous aspiration and injection of OK-432 (8 men and 12 women, mean age 63.6 years, SD 9.5). Six patients presented with flank pain, 14 presented with renal mass; renal cyst location was right, left, or bilateral sided in 9, 8, and 8 kidneys, respectively. Patients were evaluated by clinical assessment, US, or CT scan 3 months following the procedure. Complete and partial success was defined as symptom resolution with either total cyst ablation or greater than 70% reduction, respectively. Failure was defined as 30% of cyst size recurrence and/or persistent symptoms. RESULTS: Average reduction was 93.0%. Complete and partial resolution occurred in 11 (44.0%) and 13 (52.0%) cysts, respectively. One case was defined as failure, with a 64.2% size reduction from 10.9cm to 3.9cm (volume reduction rate 95.4%). Renal pain improved in all patients, regardless of complete or partial resolution. Minor complications occurred in 3 patients, 2 developed leukocytosis and 1 had mild fever (< 38.5 degrees C) following aspiration and sclerotherapy. Successful treatment was achieved with conservative measures and NSAID therapy. CONCLUSION: Percutaneous treatment of simple renal cysts with OK-432 sclerotherapy was found to be a safe, effective and minimally invasive procedure. PMID- 17461528 TI - Patterns in the diagnosis and management of benign prostatic hyperplasia in a country that does not have country-specific clinical practice guidelines. AB - PURPOSE: We have evaluated the patterns of diagnostic and treatment practices for benign prostatic hyperplasia (BPH) in a country that does not have country specific clinical practice guidelines. MATERIALS AND METHODS: Probability samples were taken from the Korean Urological Association Registry of Physicians, and randomly sampled Korean urologists were asked to complete a questionnaire. The survey explored practice characteristics and attitudes, as well as diagnostic and treatment strategies, for the management of BPH. RESULTS: Of the 850 questionnaires sent, 302 were returned, and 277 of those were included in the final analysis (response rate 32.6%). For the initial evaluation, most urologists routinely used digital rectal examinations (DRE) and urinalysis. Uroflowmetry was used 34.7% of the time. Pressure-flow studies were rarely done. Symptom assessment was used in only 46.9% of cases. In addition, a significant number (58.8%) reported that treatment decisions were not based on the symptom questionnaire. Before surgery, almost all urologists routinely used DRE, urinalysis, and prostate-specific antigen tests. Of the respondents, 55.6% and 41.9% had prescribed alpha- blockers and alpha-blockers with 5-alpha reductase inhibitors, respectively. 81.2% of urologists perceived that selective alpha blockers are different in terms of efficacy, and 82.7% felt that they differed in safety. Most respondents prescribed 5-alpha reductase inhibitors based on the prostate size. CONCLUSION: These data provide a picture of current practices regarding the management of BPH in Korea. The diagnostic and treatment practices for BPH do not follow published guidelines. Our findings ask the question "How influential are international guidelines, and do they really affect patient management in countries that do not have country-specific guidelines?" PMID- 17461527 TI - The effect of hepatitis C virus infection on insulin resistance in chronic haemodialysis patients. AB - PURPOSE: To investigate the contribution of HCV infection to insulin resistance in chronic haemodialysis patients. MATERIALS AND METHODS: The study was performed with 55 patients who were on regular haemodialysis therapy three times per week. Of the 55 patients, 34 (20 females and 14 males with an average age of 40.9 years) were anti-HCV (+) and were defined as the HCV (+) group. The remaining 21 patients (8 females and 11 males with an average age of 50 years) were negative for HCV and other viral markers and were defined as the HCV (-) group. BMI of all patients were below 27. Insulin resistance (IR) was calculated according to the HOMA formula and patients were called HOMA-IR (+) if their HOMA scores were higher than 2.5. All of the HOMA-IR (+) patients in both groups were called the HOMA-IR (+) subgroup. None of the patients had a history of drug use or any diseases that were related to insulin resistance except uremia. In both groups and the healthy control group, insulin and glucose levels were studied at three different venous serum samples taken at 5- minute intervals after 12 hours of fasting. Other individual variables were studied at venous serum samples taken after 12 hours of fasting. RESULTS: HOMA scores were (3)2.5 in 22 of 34 HCV (+) patients (64.7%) and 7 of 21HCV (-) patients (33.33%) (p=0.024). Insulin levels of HCV (+) group (13.32 +/- 9.44mIU/mL) were significantly higher than HCV (-) (9.07 +/- 7.39mIU/mL) and the control groups (6.40 +/- 4.94mIU/ mL) (p=0.039 and p=0.021 respectively). HCV (+) patients were younger (40.94 +/- 17.06 and 52.62 +/- 20.64 years, respectively) and had longer dialysis duration (7.18 +/- 3.61 and 2.91 +/- 2.69 years, respectively). Significant positive correlations of HOMA score with insulin (r=0.934, p=0.000) and fasting glucose levels (r=0.379, p=0.043) were found in the HOMA- IR (+) subgroup. Also, a significant positive correlation was found between ALT and insulin levels in the HOMA IR (+) subgroup. C-peptide levels of both HCV (+) and (-) groups were significantly higher than the control group (p < 0.001). There were not any significant correlations between HOMA score and some of the other individual variables including levels of triglyceride, ferritin, ALT, iPTH and Mg in any of the groups. CONCLUSION: In chronic haemodialysis patients; HCV infection is related to a high prevalence of insulin resistance, higher insulin and glucose levels. PMID- 17461529 TI - Black cohosh and St. John's wort (GYNO-Plus) for climacteric symptoms. AB - PURPOSE: This study was conducted to investigate the efficacy of black cohosh (Cimicifuga racemosa) and St. John's wort (Hypericum perforatum) in women with climacteric symptoms, and to assess their effects on vaginal atrophy, hormone levels, and lipid profiles. MATERIALS AND METHODS: In this double-blind randomized, placebo-controlled, multicenter study, 89 peri- or postmenopausal women experiencing climacteric symptoms were treated with St. John's wort and black cohosh extract (Gynoplus), Jin-Yang Pharm., Seoul, Korea) or a matched placebo for 12 weeks. Climacteric complaints were evaluated by the Kupperman Index (KI) initially and at 4 and 12 weeks following treatment. Vaginal maturation indices, serum estradiol, FSH, LH, total cholesterol, HDL- cholesterol, LDL-cholesterol, and triglyceride levels were measured before and after treatment. From the initial 89 participants, 77 completed the trial (42 in the Gynoplus group, 35 in the placebo group). RESULTS: Baseline characteristics were not significantly different between the two groups. Mean KI scores and hot flushes after 4 and 12 weeks were significantly lower in the Gynoplus group. Differences in superficial cell proportion were not statistically significant. HDL levels decreased in the control group from 60.20 +/- 16.37 to 56.63 +/- 12.67, and increased in the Gynoplus group from 58.32 +/- 11.64 to 59.74 +/- 10.54; this was statistically significant (p=0.04). CONCLUSION: Black cohosh and St. John's wort combination was found to be effective in alleviating climacteric symptoms and might provide benefits to lipid metabolism. PMID- 17461530 TI - Intra-synovial ropivacaine and morphine for pain relief after total knee arthroplasty: a prospective, randomized, double blind study. AB - PURPOSE: Several analgesic techniques are available for pain management after a major operation. MATERIALS AND METHODS: From December 2005 to February 2006, a prospective, double-blind study was performed involving 90 patients who had undergone a total knee arthroplasty. Patients were randomly divided into three equal groups (n=30). Demographic data, including age, height, weight, knee score, visual analogue scale (VAS), and range of flexion were evaluated preoperatively. Before wound closure, patients were given intra-synovial injections of the following solutions: patients in group I received 40mL of 300mg ropivacaine with 1:200,000 epinephrine and 5mg morphine; patients in Group II received 40mL of 300mg ropivacaine with epinephrine; and patients in Group III received 50mL normal saline as a control. All patients received an epidural patient-controlled analgesia (PCA) for 24 postoperative hours. Analgesic efficacy was evaluated using the VAS at intervals of 2, 4, 6, 12, 24, 32, 40, and 48 hours postoperatively. During this period, the side effects, the dosage of rescue analgesia required, and the range of knee flexion were recorded for each group. RESULTS: There were no significant differences among the three groups with regards to the VAS and the required dose of rescue analgesia (p > 0.05). None of the groups demonstrated significant differences in the range of knee flexion and the incidence of postoperative nausea and emesis (p > 0.05). CONCLUSION: Therefore, we found that ropivacaine, alone or with morphine, injected into the synovial tissue, along with an epidural PCA has no additional benefits in pain control after a total knee arthroplasty. PMID- 17461531 TI - Beneficial effects of thiazolidinediones on diabetic nephropathy in OLETF rats. AB - PURPOSE: Diabetic nephropathy is the most serious of complications in diabetes mellitus. Thiazolidinedione (TZD) is thought to ameliorate diabetic nephropathy; however, the mechanism underlying this effect has not been elucidated. We hypothesized that the vascular endothelial growth factor (VEGF) participates in the pathogenesis of diabetic nephropathy and that TZD may be beneficial for the treatment of diabetic nephropathy because of the effect it has on VEGF. MATERIALS AND METHODS: 23 Otsuka- Long-Evans-Tokushima-Fatty (OLETF) rats and eight control Long-Evans-Tokushima-Otsuka (LETO) rats were divided into the following four groups: LETO group, control OLETF group, pioglitazone treated group (10mg/ kg/day), and rosiglitazone treated group (3mg/kg/day). RESULTS: A progressive increase in urinary protein excretion was observed in the diabetic rats. Glomerular VEGF expression in the control OLETF rats was significantly higher than in the control LETO rats. However, there was a significant reduction in both the glomerular VEGF expression and the VEGF mRNA levels after treatment with pioglitazone and rosiglitazone. The twenty-four hour urine protein levels were significantly decreased in both groups of the treated OLETF rats. CONCLUSION: These results suggest that TZD may have beneficial effects on diabetic nephropathy by reducing the VEGF expression. PMID- 17461532 TI - Induction of PPAR gamma mRNA and protein expression by rosiglitazone in chronic cyclosporine nephropathy in the rat. AB - PURPOSE: We recently reported that rosiglitazone (RGTZ), a peroxisome proliferator-activated receptor gamma (PPARgamma) agonist, has a protective effect against cyclosporine (CsA)- induced renal injury. Here we report the effect of RGTZ on peroxisome proliferator-activated receptor gamma (PPARgamma) expression in an experimental model of chronic cyclosporine (CsA) nephropathy. MATERIALS AND METHODS: Chronic CsA nephropathy was induced in Sprague-Dawley rats by administering CsA (15mg/kg per day) for 28 days, and control rats were treated with vehicle (VH group, olive oil 1mL/kg per day) for 28 days. RGTZ (3mg/kg) was concurrently administered via gavage to the CsA and VH groups. Expression of PPARgamma mRNA and protein was evaluated with RT-PCR, immunohistochemistry, and immunoblotting. RESULTS: PPARgamma mRNA expression was similar to the level of PPARgamma protein constitutively expressed in the kidneys of the VH treated rats, with expression in the glomerular epithelial, distal tubular cells, and collecting tubular cells. PPARgamma protein expression in CsA-treated rat kidneys was significantly less than in the VH group. However, concomitant administration of RGTZ restored PPARgamma protein expression in the kidneys of the CsA- reated rats. CONCLUSION: Exogenous administration of RGTZ treatment upregulates PPARgamma expression and that this mechanism may play a role in protecting against CsA-induced renal injury. PMID- 17461533 TI - Anticonvulsant hypersensitivity syndrome associated with Epstein-Barr virus reactivation. AB - We describe a 59-year-old female with severe anticonvulsant hypersensitivity syndrome (AHS) associated with Epstein- Barr virus (EBV) infection. The causative drug was speculated to be carbamazepine. Recurrent EBV infection was demonstrated by the presence of anti-EBV early antigen IgM antibodies and anti-EBV nuclear antigen IgG antibodies. To our knowledge, only one case of drug hypersensitivity syndrome (DHS) associated with EBV has been reported in the English- language literature. Our case is the second report of EBV-associated DHS, which suggests that EBV infection may contribute to the pathogenesis of AHS in a few patients. PMID- 17461534 TI - Acute Marchiafava-Bignami disease: diffusion-weighted MRI in cortical and callosal involvement. AB - Marchiafava-Bignami disease (MBD) is a fatal disorder characterized by demyelination of the corpus callosum. MRI, suggestive of corpus callosum demyelination with associated white matter involvement in both cerebral hemispheres, indicates a diagnosis of MBD. In this case, MR diffusion-weighted findings taken at an acute stage of MBD revealed lesions not only in the corpus callosum but also in the cerebral cortex. Lower apparent diffusion coefficient values of the corpus callosum and cortical lesions were associated with poor clinical outcome. PMID- 17461535 TI - Photodynamic therapy for subretinal new vessels. AB - Photodynamic therapy (PDT) involves the induction of endothelial cell death or occlusion of blood vessels. On the basis of this mechanism of action, PDT is used in the treatment of predominant classic choroidal neovascularization (CNV), if the classic component is over 50%, and in myopic CNV. This study describes 2 cases of distinctive, abnormal, large, subretinal new vessels that are thought to have originated from the choroids. Diminishment of the new vessels was observed following treatment with PDT. PMID- 17461536 TI - Penetrating retrobulbar orbital foreign body: a transcranial approach. AB - We report the successful removal of a retrobulbar foreign body using a transcranial approach in a 63-year-old patient with a penetrating injury to the left eye. Initial ocular examination revealed a corneoscleral laceration, hyphema, a traumatic cataract, and vitreous hemorrhage. Visual acuity consisted only of the perception of hand motion. Computed tomography demonstrated an orbital foreign body in the retrobulbar area. Emergency corneoscleral suturing, phacoemulsification of the cataract, and vitrectomy with posterior vitreous detachment were performed. Fifteen days after the emergency operation, we successfully removed the orbital foreign body using a transcranial approach, although the foreign body was very close to the optic nerve. On fundus examination 6 months later, a white, fibrous lesion was seen inferior to the optic disc, and the corrected visual acuity was 20/30. These positive results may be due to the complete vitrectomy at the correct time performed by a retina specialist and the minimal pressure on the eyeball while removing the foreign body, which resulted from the use of a transcranial approach. PMID- 17461537 TI - Bupivacaine induced cardiac toxicity mimicking an acute non-ST segment elevation myocardial infarction. AB - Bupivacaine is widely used as a local anesthetic. Central nervous system (CNS) and cardiovascular toxicity are well known side effects. However, there has been no report of bupivacaine-induced myocardial injury. We present a case of bupivacaine cardiac toxicity mimicking an acute non-ST segment elevation myocardial infarction, which was eventually diagnosed as bupivacaine-induced cardiac toxicity without CNS toxicity. As soon as a healthy young woman at a private clinic was given a spinal anesthesia of 6mg bupivacaine for hemorrhoidectomy, she developed arrhythmia and hypotension. She was transferred to our emergency room. There was an accelerated idioventricular rhythm with ST segment depression on electrocardiogram, coarse breathing sounds with rales on whole lung field and a butterfly sign on the chest radiograph. 2D transthoracic echocardiography (TTE) revealed reduced left ventricle systolic ejection fraction (approximately 27%). There was regional wall motion abnormality of the left ventricle on 2D TTE and the cardiac marker was increased. We diagnosed the patient as having acute non-ST segment elevation myocardial infarction but her impaired cardiac function improved gradually. On the seventh day from admission, there was a complete spontaneous recovery of cardiac function, and coronary angiography revealed a normal coronary artery. Therefore, we firmly believe that bupivacaine directly injures the cardiac cell. PMID- 17461538 TI - Chronic expanding hematoma of the thorax. AB - We report the first case in Korea of a chronic expanding hematoma, which presented as a huge mass in the pleural cavity. A 67-year-old woman exhibiting a slowly-expanding intrathoracic mass, as revealed by a chest radiograph, was admitted to our hospital. The patient had undergone a pneumonectomy 37 years earlier during treatment for pulmonary tuberculosis. Computed tomography revealed a huge mass in her right hemithorax. The differential diagnosis of this mass included chronic empyema combined with a malignancy, such as lymphoma or a soft tissue sarcoma. The tumor, which was classified as an encapsulated chronic hematoma, was removed surgically. Samples sent for histopathological and microbiological analysis revealed no evidence of neoplasia or infection. The patient was finally diagnosed with a chronic expanding hematoma of the thorax. This case is particularly rare due to the patient's development of a very large mass after undergoing treatment for tuberculosis more than 30 years earlier. PMID- 17461539 TI - Robot-assisted laparoscopic radical prostatectomy: four cases. AB - The role of the da Vinci robot is being defined in minimally invasive urologic surgery. Robot-assisted laparoscopic radical prostatectomy (rLRP) has emerged as a feasible treatment option for patients with organ-confined prostate cancer. We performed the first four rLRPs on four prostate cancer patients in the Republic of Korea. This is a report of its techniques and outcomes. In all four cases, the surgery was successfully completed with a mean operative time of 392.5 minutes. The mean estimated blood loss was 312.5mL, and catheterization lasted 14 to 21 days. There were no major intraoperative or postoperative complications. The mean hospital stay was 11 days. The rLRP is a safe and feasible approach. It will become one of the standard options for the management of localized prostate cancer. PMID- 17461540 TI - Harnessing the sol-gel process for the assembly of non-silicate mesostructured oxide materials. AB - Mesostructured non-silicate oxides, with well-defined organization on the 2-50 nm size scale, may play a pivotal role in advancing vital disciplines such as catalysis, energy conversion, and biotechnology. Herein, we present selected methodologies for utilizing the sol-gel process, in conjunction with organic directed assembly, to synthesize a variety of mesostructured oxides. The nature of the inorganic precursor is critical for this process. We discuss the development of general routes for yielding stable, nanoscopic, hydrophilic, inorganic precursors compatible with organic co-assembly. In particular, we highlight the use and characterization of organic-acid-modified transition metal oxide sol-gel precursors that allow for the synthesis and processing of designer mesostructured oxides such as titania hybrids for optical applications and porous multicomponent metal oxides useful for catalysis. PMID- 17461541 TI - Monooxygenase activity of type 3 copper proteins. AB - The molecular mechanism of the monooxygenase (phenolase) activity of type 3 copper proteins has been examined in detail both in the model systems and in the enzymatic systems. The reaction of a side-on peroxo dicopper(II) model compound ( A) and neutral phenols proceeds via a proton-coupled electron-transfer (PCET) mechanism to generate phenoxyl radical species, which collapse each other to give the corresponding C-C coupling dimer products. In this reaction, a bis(mu oxo)dicopper(III) complex ( B) generated by O-O bond homolysis of A is suggested to be a real active species. On the other hand, the reaction of lithium phenolates (deprotonated form of phenols) with the same side-on peroxo dicopper(II) complex proceeds via an electrophilic aromatic substitution mechanism to give the oxygenated products (catechols). The mechanistic difference between these two systems has been discussed on the basis of the Marcus theory of electron transfer and Hammett analysis. Mechanistic details of the monooxygenase activity of tyrosinase have also been examined using a simplified enzymatic reaction system to demonstrate that the enzymatic reaction mechanism is virtually the same as that of the model reaction, that is, an electrophilic aromatic substitution mechanism. In addition, the monooxygenase activity of the oxygen carrier protein hemocyanin has been explored for the first time by employing urea as an additive in the reaction system. In this case as well, the ortho hydroxylation of phenols to catechols has been demonstrated to involve the same ionic mechanism. PMID- 17461542 TI - Metal unsaturation and ligand hemilability in Suzuki coupling. AB - The combinative and complementary use of a hemilabile difunctional ligand on a metal, notably palladium, that is coordinatively and electronically unsaturated has led to the isolation of a string of unexpected low-valent complexes that are structurally intriguing. The ligands of interest are primarily ferrocenes functionalized by [P,N] and [P,O] donors. The characterization of these active Suzuki catalysts, which support sp(2)-sp(2) couplings, give valuable insights into the key Suzuki intermediates such as those arising from the reductive elimination, transmetallation, and oxidative addition steps. In this Account, we shall review and discuss our recent results in relation to selected developments in other laboratories. PMID- 17461543 TI - Advanced solid state NMR techniques for the characterization of sol-gel-derived materials. AB - A large array of advanced solid state NMR (nuclear magnetic resonance) techniques is presented in the frame of the structural characterization of sol-gel-derived materials. These techniques include the pertinent detection of (17)O chemical shifts, MAS (magic angle spinning) J spectroscopy in the solid state, high resolution (1)H spectroscopy, heteronuclear and homonuclear D (dipolar)-derived multidimensional correlation experiments, and first-principles calculations of NMR parameters. This spectroscopic approach is suitable for the in-depth description of multicomponent sol-gel derivatives, crystalline and amorphous biocompatible silicophosphates, Al-O-P clusters, and templated porous materials. It offers unique perspectives for the description of the hybrid interfaces in terms of chemical and spatial connectivities. PMID- 17461544 TI - Nonaqueous sol-gel routes to metal oxide nanoparticles. AB - Sol-gel routes to metal oxide nanoparticles in organic solvents under exclusion of water have become a versatile alternative to aqueous methods. In comparison to the complex aqueous chemistry, nonaqueous processes offer the possibility of better understanding and controlling the reaction pathways on a molecular level, enabling the synthesis of nanomaterials with high crystallinity and well-defined and uniform particle morphologies. The organic components strongly influence the composition, size, shape, and surface properties of the inorganic product, underlining the demand to understand the role of the organic species at all stages of these processes for the development of a rational synthesis strategy for inorganic nanomaterials. PMID- 17461545 TI - Peptide beacons: a new design for polypeptide-based optical biosensors. AB - Both epitope mapping and other in vitro selection techniques produce short polypeptides that tightly and specifically bind to any of a wide range of macromolecular targets. Here, we demonstrate a potentially general means of converting such polypeptides into optical biosensors. The sensing architecture we have developed, termed peptide beacons, is based on the observation that, whereas short peptides are almost invariably unfolded and highly dynamic, they become rigid when complexed to a macromolecular target. Using this effect to segregate a long-lived fluorophore from an electron transfer based, contact quencher (both covalently attached to the peptide), we have produced a robust optical sensor for anti-HIV antibodies. The binding-induced segregation of the fluorophore-quencher pair produces a 6-fold increase in sensor emission, thus allowing us to readily detect as low as approximately 250 pM of the target antibody. Because the sensor is based on binding-induced folding and a visible-light fluorophore, it is sufficiently selective to work directly in complex, contaminant-ridden samples such as saliva and blood. PMID- 17461546 TI - Acid-degradable protein delivery vehicles based on metathesis chemistry. AB - In this communication we demonstrate that acyclic diene metathesis (ADMET) polymerization is a powerful methodology for the synthesis of acid-degradable polymers based on polyketals and polyacetals. Ten new polyketals and polyacetals were synthesized, using ADMET, and a polyacetal based on anthracene aldehyde was identified, which had the physical properties needed for microparticle formulation. The antioxidant protein catalase was encapsulated into microparticles, formulated from this polyacetal, using a double emulsion procedure, and cell culture studies demonstrated that these microparticles dramatically improved the ability of catalase to scavenge hydrogen peroxide produced by macrophages. We anticipate numerous applications of ADMET for the synthesis of acid-degradable polymers based on its excellent tolerance toward functional groups and ease of synthesis. PMID- 17461547 TI - Enhancing the production of Fc fusion protein in fed-batch fermentation of Pichia pastoris by design of experiments. AB - This study focuses on the feasibility of producing a therapeutic Fc fusion protein in Pichia pastoris (P. pastoris) and presents an optimization design of experiment (DOE) strategy in a well-defined experimental space. The parameters examined in this study include pH, temperature, salt supplementation, and batch glycerol concentration. The effects of these process conditions were captured by statistical analysis focusing on growth rate and titer responses. Batch medium and fermentation conditions were also investigated prior to the DOE study in order to provide a favorable condition to enable the production of this Fc fusion protein. The results showed that approximately 373 mg/L of the Fc fusion protein could be produced. The pH was found to be particularly critical for the production of this Fc fusion protein. It was significantly higher than the conventional, recommended pH for P. pastoris fermentation. The development of this process shows that protein production in P. pastoris is protein specific, and there is not a set of pre-defined conditions that can work well for all types of proteins. Thorough process development would need to be performed for every type of protein in order for large-scale production in P. pastoris to be feasible. PMID- 17461548 TI - Comparison of spiculogenesis in in vitro ADCP-primmorph and explants culture of marine sponge Hymeniacidon perleve with 3-TMOSPU supplementation. AB - This study aims to test the feasibility of introducing functional chemical groups into biogenic silica spicules by examining the effect of supplementing a silican coupler [3-(trimethoxysilyl)propyl]urea (3-TMOSPU) as silica source in the cultures of archaeocytes-dominant-cell-population (ADCP) primmorphs and explants of the marine sponge Hymeniacidon perleve. Analysis by Fourier Transform Infrared Spectroscopy (FT-IR) confirmed that the organic group in 3-TMOSPU was introduced into silica spicules. By comparing ADCP-primmorph cultures when supplemented with Na2SiO3, 3-TMOSPU supplementation showed no notable effect on the primmorphs development and cell locomotion behaviors. A decline in silicatein expression quantified by real-time RT-PCR was, however, observed during spiculogenesis. The decline was slower for the 3-TMOSPU group whereas significantly fewer spicules were formed. When sponge papillae explants were cultured, 3-TMOSPU supplementation had no negative effect on sponge growth but inhibited the growth biofouling of the diatom Nitzschia closterium. By monitoring the detectable Si concentration, it seemed that 3-TMOSPU was converted by the sponge and its conversion was related to spiculogenesis. Analysis of spicule dimensional changes indicated that the inhibition of spiculogenesis by 3-TMOSPU supplementation was less in ADCP-primmorphs culture due to lower 3-TMOSPU/detectable Si ratio in the media. PMID- 17461549 TI - Bioreactor production of recombinant herpes simplex virus vectors. AB - Serotypical application of herpes simplex virus (HSV) vectors to gene therapy (type 1) and prophylactic vaccines (types 1 and 2) has garnered substantial clinical interest recently. HSV vectors and amplicons have also been employed as helper virus constructs for manufacture of the dependovirus adeno-associated virus (AAV). Large quantities of infectious HSV stocks are requisite for these therapeutic applications, requiring a scalable vector manufacturing and processing platform comprised of unit operations which accommodate the fragility of HSV. In this study, production of a replication deficient rHSV-1 vector bearing the rep and cap genes of AAV-2 (denoted rHSV-rep2/cap2) was investigated. Adaptation of rHSV production from T225 flasks to a packed bed, fed-batch bioreactor permitted an 1100-fold increment in total vector production without a decrease in specific vector yield (pfu/cell). The fed-batch bioreactor system afforded a rHSV-rep2/cap2 vector recovery of 2.8 x 10(12) pfu. The recovered vector was concentrated by tangential flow filtration (TFF), permitting vector stocks to be formulated at greater than 1.5 x 10(9) pfu/mL. PMID- 17461550 TI - Calcium alginate bead immobilization of cells containing tyrosine ammonia lyase activity for use in the production of p-hydroxycinnamic acid. AB - An Escherichia coli catalyst with tyrosine ammonia lyase activity (TAL) has been stabilized for repeated use in batch conversions of high tyrosine solids to p hydroxycinnamic acid (pHCA). The TAL biocatalyst was stabilized by controlling the reaction pH to 9.8 +/- 0.1 and immobilizing the cells within a calcium alginate matrix that was cross-linked with glutaraldehyde and polyethyleneimine (GA/PEI). We found a GA range where the bead-encapsulated TAL was not inactivated, and the resulting cross-linking provided the beads with the mechanical stability necessary for repeated use in consecutive batch reactions with catalyst recycle. The GA/PEI calcium alginate TAL catalyst was used in 41 1 L batch reactions where 50 g L(-1) tyrosine was converted to 39 +/- 4 g L(-1) pHCA in each batch. The practical usefulness and ease of this process was demonstrated by scaling up the TAL bead immobilization and using the immobilized TAL catalyst in four 125-L bioconversion reactions to produce over 12 kg of purified pHCA. PMID- 17461551 TI - Improved production of biosurfactant with newly isolated Pseudomonas aeruginosa S2. AB - An indigenous strain Pseudomonas aeruginosa S2 (P. aeruginosa S2), isolated from diesel-contaminated soil, produced extracellular surface-active material identified as rhamnolipid. Due to its excellent surface activity, rhamnolipid is known to be well-suited for stimulating the bioremediation efficiency of oil contaminated sites. To improve production yield of rhamnolipid with P. aeruginosa S2, various carbon and nitrogen sources were screened to select favorable ones leading to better biosurfactant production yield. It was found that using 4% glucose could attain better rhamnolipid yield, while 50 mM NH4NO3 appeared to be the most preferable nitrogen source. Meanwhile, the effect of carbon to nitrogen ratio (C/N ratio) on rhamnolipid yield was also investigated, and the optimal C/N ratio was identified as approximately 11.4. Moreover, response surface methodology (RSM) was applied to optimize the trace element concentration for rhamnolipid production. Results from two-level design indicate that concentrations of MgSO4 and FeSO4 were the most significant factors affecting rhamnolipid production. Using steepest ascent method and RSM analysis, an optimal medium composition was determined, giving a rhamnolipid production yield of 2.37 g/L in 100 h at 37 degrees C and 200 rpm agitation. Scale-up production of rhamnolipid in a well-controlled 5 L jar fermentor using the optimal medium and operating condition (at 37 degrees C and pH 6.8) further elevated the biosurfactant production yield to 5.31 g/L (in 97 h), which is over 2-fold higher than the best results obtained from shake-flask tests. PMID- 17461552 TI - Mass spectrometric analysis for high molecular weight synthetic polymers using ultrasonic degradation and the mechanism of degradation. AB - We have investigated ultrasonic degradations of poly(ethylene oxide) (PEG) and poly(methyl methacrylate) (PMMA) in aqueous media by means of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS). The ultrasonic degradation of polymers was monitored as a function of ultrasonication duration to examine the structural details of ultrasonic degradation polymers. PEG solution ultrasonication produced five types of oligomers (M approximately 1000 Da) with different end groups, irrespective of the initial average molecular masses (M=2, 6, 20, and 2000 kDa). Several degradation pathways with free radical reactions have been suggested to explain these degradation products: the ultrasonic degradation of PEG is initiated by breaking of the C-O bond in the PEG chain, generating polymeric radicals with two terminal groups, i.e., X*( approximately CH2CH2*) and Y*( approximately CH2CH2O*), followed by termination with extraction or release of a hydrogen atom. However, PMMA (M=1630 Da) ultrasonication generated only one type of degradation oligomer, which has a hydrogen group at both ends, the same as that of the original oligomer. It has been suggested that the presence of the radical terminal groups X*( approximately CH2*) and Y*( approximately (CH3)CCOO(CH3)C*) is due to selective C-C bond breaking in the chain during the ultrasonic degradation of PMMA. The MALDI-TOFMS combined with the ultrasonic degradation technique (UD/MALDI-TOFMS) developed in this study could be extended to the analysis of synthetic polymer structures with high molecular weights. PMID- 17461553 TI - Structure of the catalytic domain of human polo-like kinase 1. AB - Polo-like kinase 1 (Plk1) is an attractive target for the development of anticancer agents due to its importance in regulating cell-cycle progression. Overexpression of Plk1 has been detected in a variety of cancers, and expression levels often correlate with poor prognosis. Despite high interest in Plk1 targeted therapeutics, there is currently no structure publicly available to guide structure-based drug design of specific inhibitors. We determined the crystal structures of the T210V mutant of the kinase domain of human Plk1 complexed with the nonhydrolyzable ATP analogue adenylylimidodiphosphate (AMPPNP) or the pyrrolo-pyrazole inhibitor PHA-680626 at 2.4 and 2.1 A resolution, respectively. Plk1 adopts the typical kinase domain fold and crystallized in a conformation resembling the active state of other kinases. Comparison of the kinetic parameters determined for the (unphosphorylated) wild-type enzyme, as well as the T210V and T210D mutants, shows that the mutations primarily affect the kcat of the reaction, with little change in the apparent Km for the protein or nucleotide substrates (kcat = 0.0094, 0.0376, and 0.0049 s-1 and Km(ATP) = 3.2, 4.0, and 3.0 microM for WT, T210D, and T210V, respectively). The structure highlights features of the active site that can be exploited to obtain Plk1 specific inhibitors with selectivity over other kinases and Plk isoforms. These include the presence of a phenylalanine at the bottom of the ATP pocket, combined with a cysteine (as opposed to the more commonly found leucine) in the roof of the binding site, a pocket created by Leu132 in the hinge region, and a cluster of positively charged residues in the solvent-exposed area outside of the adenine pocket adjacent to the hinge region. PMID- 17461554 TI - Bromine enrichment in the near-surface region of Br-doped NaCl single crystals diagnosed by Rutherford backscattering spectrometry. AB - Bromine released from sea-salt aerosols and seawater ice is known for its high chemical reactivity. Previous studies have suggested that its availability to the gas-phase could be enhanced by segregation processes increasing Br concentration on the aerosol surface as compared to the bulk. However, little is known about the composition within the near-surface region, that is, the outermost approximately 100 monolayers. We used Rutherford backscattering spectrometry (RBS) to measure Br concentration profiles to a depth of about 750 nm of Br-doped NaCl single crystals to characterize the thermodynamics and kinetics of Br segregation to the near-surface region in moist air. These experiments were carried out on cleavage planes of melt-grown and of annealed solution-grown crystals at room temperature and relative humidities (RH) too low for formation of a stable liquid phase. Segregation of Br was below the detection limit on melt grown crystals with Br/Cl = 0.01. In the case of annealed solution-grown crystals with Br/Cl = 0.002, average segregations of (0.24 +/- 0.11) x 10(15) and (0.42 +/ 0.12) x 10(15) Br atoms cm-2 were observed at 50% and 65% RH, respectively. No segregation was found at 20% RH. The observed Br segregation can be explained by the formation of an adsorbed liquid layer (depending on crystal surface properties and relative humidity) and preferential, diffusion-limited dissolution of Br into this layer according to the partition coefficient of Br between aqueous and solid NaCl. The thickness of the adsorbed liquid layer, which depends on crystal surface geometry and on relative humidity, can be estimated to range from 4 to at most 59 nm on the basis of measured Br concentrations and partition coefficients. Applying this concept of partitioning to natural sea salt suggests a Br/Cl molar ratio of up to 0.2 in adsorbed surface water of crystallized natural aerosol particles compared to about 0.0015 in seawater. This would have a major impact on heterogeneous reactions on sea-salt particles under dry conditions such as in the freeze-dried Arctic boundary layer. PMID- 17461555 TI - Electronic excitations of glycine, alanine, and cysteine conformers from first principles calculations. AB - The electronic and optical properties are studied for three conformers of amino acid molecules using gradient-corrected (spin-) density functional theory within a projector-augmented wave scheme and the supercell method. We investigate single particle excitations such as ionization energies and electron affinities as well as pair excitations. By comparing eigenvalues resulting from several local and nonlocal energy functionals, the influence of treatment of exchange and correlation is demonstrated. The excitations are described within the Delta-self consistent field method with an occupation number constraint to obtain excitation energies and Stokes shifts. The results are used to also discuss the optical absorption properties. In contrast to the lowest single- and two-particle excitation energies, remarkable changes are found in absorption spectra in dependence on the conformation of the molecule geometry. PMID- 17461556 TI - Fast approaches for molecular polarizability calculations. AB - Molecular polarizability of a molecule characterizes the capability of its electronic system to be distorted by the external field, and it plays an important role in modeling many molecular properties and biological activities. In this paper, a set of fast empirical models have been developed to predict molecular polarizability using two types of approaches. The first type of approaches is based on Slater's rules of calculating the effective atomic nuclear shielding constants. The best model (model 1A) of this category has achieved an average unsigned error (AUE), root-mean square error (RMSE), and average percent error (APE) of 2.23 au, 3.29 au, and 2.77%, respectively. The second type of model is based on an additive hypothesis of molecular polarizability. Five models have been constructed using different schemes of atom types. The best model that applies 14 atom types, model 2e, achieves AUE, RMSE, and APE of 0.99 au, 1.48 au, and 1.24%, respectively. This performance is much better than those of the models purely based upon chemical composition (model 2A and the Bosque and Sales model), for which errors are about 2-fold higher. It is expected that both model 1A and model 2E will have broad applications in QSAR and QSPR studies. PMID- 17461557 TI - Oxygen discharge and post-discharge kinetics experiments and modeling for the electric oxygen-iodine laser system. AB - Laser oscillation at 1315 nm on the I(2P1/2)-->I(2P3/2) transition of atomic iodine has been obtained by a near resonant energy transfer from O2(a1Delta) produced using a low-pressure oxygen/helium/nitric oxide discharge. In the electric discharge oxygen-iodine laser (ElectricOIL) the discharge production of atomic oxygen, ozone, and other excited species adds levels of complexity to the singlet oxygen generator (SOG) kinetics which are not encountered in a classic purely chemical O2(a1Delta) generation system. The advanced model BLAZE-IV has been introduced to study the energy-transfer laser system dynamics and kinetics. Levels of singlet oxygen, oxygen atoms, and ozone are measured experimentally and compared with calculations. The new BLAZE-IV model is in reasonable agreement with O3, O atom, and gas temperature measurements but is under-predicting the increase in O2(a1Delta) concentration resulting from the presence of NO in the discharge and under-predicting the O2(b1Sigma) concentrations. A key conclusion is that the removal of oxygen atoms by NOX species leads to a significant increase in O2(a1Delta) concentrations downstream of the discharge in part via a recycling process; however, there are still some important processes related to the NOX discharge kinetics that are missing from the present modeling. Further, the removal of oxygen atoms dramatically inhibits the production of ozone in the downstream kinetics. PMID- 17461558 TI - Intensity of d-d symmetry-forbidden electronic transition in Cr(CO)6. AB - Absolute absorption intensities (oscillator strengths) are calculated for the d-d symmetry-forbidden transition in hexacarbonyl chromium. The vibronic coupling mechanism is taken into account in a way that represents an alternative to the traditional perturbative approach of Herzberg and Teller. In the so-called direct method, the electronic transition moment is directly expanded in a power series of the vibrational normal coordinates of suitable symmetry. In the present case, i.e., d-d ligand field transitions, or more specifically (1)A(1g) --> (1)T(1g) and (1)A(1g) --> (1)T(2g) transitions, the dipole selection rule is broken by vibronic interaction induced by normal modes that transform like T(1u) and T(2u) representations of the O(h) group. An analysis of the relative importance of normal modes in promoting electronic transitions is carried out. PMID- 17461559 TI - Ab initio study of the electronic structure and bonding of aluminum nitride. AB - For the diatomic aluminum nitride (AlN), we have constructed potential energy curves for 45 states employing multi-reference variational methods and quantitative basis sets. Thirty-six states are relatively strongly bound, five present local minima, and four are of repulsive nature. Almost all states are of intense multi-reference character rendering their calculation and interpretation quite problematic. Our tentative assignment of the ground state is 3Pi, while a 3Sigma- state is above by less than 1 kcal/mol. Our best estimate for the binding energy of the X3Pi state is D0 = 56.0 +/- 0.5 kcal/mol at re = 1.783 A, in good agreement with the experimental values of D = 66 +/- 9 kcal/mol and re = 1.7864 A. The binding energy of the A3Sigma- state is very similar to the X state because they both correlate to the ground-state atoms, but the bond distance of the former is 0.13 A longer. The first seven states can be tagged as follows: X3Pi, A3Sigma-, a1Sigma+, b1Pi, c1Delta, B3Sigma+, and d1Sigma+, a rather definitive order with the exception of X and A states. PMID- 17461560 TI - Structures of water octamers (H2O)8: exploration on ab initio potential energy surfaces by the scaled hypersphere search method. AB - The potential energy surface (PES) of water octamers has been explored by the scaled hypersphere search method. Among 164 minima on the PES (based on MP2/6 311++G(3df,2p)//B3LYP/6-311+G(d,p) calculations), the cubic structure with D2d symmetry has been confirmed to be the global minimum. In a thermodynamic simulation using these 164 structures, the cubic structure with S4 symmetry has the highest population at low temperature, though double rings can become dominant as temperature going up, in good accord with a recent Monte Carlo simulation using an empirical potential. A transition temperature from cubic to noncubic has significantly been underestimated when potential energy data of B3LYP/6-311+G(d,p) calculations are employed in the simulation. This serious discrepancy between the MP2 and the B3LYP results suggests an importance of dispersion interactions for discussions on thermodynamics of water octamers. PMID- 17461561 TI - A theoretical study of the photodetachment and intramolecular hydrogen-bonding energies of hydrogen maleate anions. AB - Three low-lying conformers of the hydrogen maleate anions (HMAs) regarding cis HMA(HB) having the O-...HO intramolecular hydrogen bond (HB), cis-HMA(nHB) without the HB, and trans-HMA are studied by density functional theory (B3LYP) combined with natural bond orbital (NBO) and atoms-in-molecules (AIM) analyses. The photoelectron spectra of cis- and trans-HMA conformers recorded by Woo et al. (J. Phys. Chem. A 2005, 109, 10633) are reassigned on the basis of the present electron propagator theory calculations, indicating the significant energy differences between the Dyson orbitals and canonical molecular orbitals due to the electron-correlation and orbital relaxation effects considered in the electron propagator theory. The NBO associated with the natural resonance theory analyses and AIM electron topological study show that the strong O-...HO in cis HMA(HB) has the remarkable characteristics of three-center four-electron hyperbond, and the bonding strength of ca. 30 kcal/mol is recommended with the reference calculations of the HO-...HOH complex. The further calculations for the microhydrated cis-HMA(HB) clusters indicate that the O-...HO bonding strength decreases in water solution. PMID- 17461562 TI - Zinc oxide prepared by homogeneous hydrolysis with thioacetamide, its destruction of warfare agents, and photocatalytic activity. AB - Zinc sulfide (ZnS) nanoparticles were prepared by homogeneous hydrolysis of zinc sulfate and thioacetamide (TAA) at 80 degrees C. After annealing at a temperature above 400 degrees C in oxygen atmosphere, zinc oxide (ZnO) nanoparticles were obtained. The ZnS and ZnO nanoparticles were characterized by X-ray diffraction (XRD), scanning electron microscopy (SEM), high-resolution transmission electron microscopy (HRTEM), selected area electron diffraction (SAED), and Brunauer Emmett-Teller (BET)/Barrett-Joyner-Halenda (BJH) methods were used for surface area and porosity determination. The photocatalytic activity of as-prepared zinc oxide samples was determined by decomposition of Orange II dye in aqueous solution under UV irradiation of 365 nm wavelength. Synthesized ZnO were evaluated for their non-photochemical degradation ability of chemical warfare agents to nontoxic products. PMID- 17461563 TI - Wave packet dynamics in triplet states of Na2 attached to helium nanodroplets. AB - The dynamics of vibrational wave packets excited in Na2 dimers in the triplet ground and excited states is investigated by means of helium nanodroplet isolation (HENDI) combined with femtosecond pump-probe spectroscopy. Different pathways in the employed resonant multiphoton ionization scheme are identified. Within the precision of the method, the wave packet dynamics appears to be unperturbed by the helium droplet environment. PMID- 17461564 TI - Electronic structure and bonding in actinyl ions and their analogs. AB - This Feature Article seeks to present the current state of knowledge, both experimental and theoretical, of the electronic structure and bonding in actinyl ions and related species, such as the isoelectronic imido compounds as well as in linear triatomic actinide molecules of the type X-An-Y. PMID- 17461565 TI - Identification of natural-product-derived inhibitors of 5-lipoxygenase activity by ligand-based virtual screening. AB - A natural product collection and natural-product-derived combinatorial libraries were virtually screened for potential inhibitors of human 5-lipoxygenase (5-LO) activity. We followed a sequential ligand-based approach in two steps. First, similarity searching with a topological pharmacophore descriptor (CATS 2D method) was performed to enable scaffold-hopping. Eighteen compounds were selected from a virtual hit list of 430 substances, which had mutual pharmacophore features with at least one of 43 known 5-LO inhibitors that served as query structures. Two new chemotypes exhibited significant activity in a cell-based 5-LO activity assay. The two most potent molecules served as seed structures for a second virtual screening round. This time, a focused natural-product-derived combinatorial library was analyzed by different ligand-based virtual screening methods. The best molecules from the final set of screening candidates potently suppressed 5 LO activity in intact cells and may represent a novel class of 5-LO inhibitors. The results demonstrate the potential of natural-product-derived screening libraries for hit and lead structure identification. PMID- 17461566 TI - Discovery of 3-piperidinyl-1-cyclopentanecarboxamide as a novel scaffold for highly potent CC chemokine receptor 2 antagonists. AB - Introduction of ring restrictions to a linear aminobutyramide CC chemokine receptor 2 (CCR2) antagonist lead (2) led to the discovery of a 1,3-disubstituted cyclopentane scaffold with enhanced hCCR2 receptor binding and antagonist activity. (1S,3R)-N-[3,5-Bis(trifluoromethyl)benzyl]-1-methyl-3-[(1R,3'R)-methyl 1'H-spiro[indene-1,4'-piperidin]-1'-yl]cyclopentanecarboxamide (16) had IC50 of 1.3 nM (binding) and 0.45 nM (functional chemotaxis) against hCCR2. It also showed activity against the mouse CCR2 receptor with an IC50 of 130 nM. Compound 16 is selective against other chemokine receptors, including CCR5 ( approximately 500-fold). PMID- 17461567 TI - The influence of secondary interactions during the formation of polyelectrolyte multilayers: layer thickness, bound water and layer interpenetration. AB - With X-ray and neutron reflectivity, the structure and composition of polyelectrolyte multilayers from poly(allyl amine) (PAH) and poly(styrene sulfonate) (PSS) are studied as function of preparation conditions (salt concentration and solution temperature, T). The onset of a temperature effect occurs at 0.05 M NaCl (Debye length approximately 1 nm). At 1 M salt, the film thickness increases by a factor of 3 on heating the deposition solution from 5 to 60 degrees C. The PAH/PSS bilayer thickness is independent of the kind of salt (NaCl or KCl), yet its composition is different (more bound water for NaCl). At low T, the internal roughness is 33% of the bilayer thickness; it increases to 60% at high T. The roughening is accompanied by a total loss of bound water. At which temperature the roughening starts is a function of the kind of salt (50 degrees C for NaCl and 35 degrees C for KCl). The strong temperature dependence and the eventual loss of bound water molecules may be attributed to the hydrophobic force; however, there is an isotope effect, since the loss of bound water is less pronounced in the deuterated layers. PMID- 17461568 TI - Linking phase behavior and reversible colloidal aggregation at low concentrations: simulations and stochastic mean field theory. AB - We have studied the link between the kinetics of clustering and the phase behavior of dilute colloids with short range attractions of moderate strength. This was done by means of computer simulations and a theoretical kinetic model originally developed to deal with reversible colloidal aggregation. Three different regions of the phase diagram were accessed. For weak attractions, a gas phase of small clusters in equilibrium forms in the system. For intermediate attractions, the system undergoes liquid-gas separation, which is signatured by the formation of a few large droplike aggregates, a gas phase of small clusters, and an overall kinetics where a few seeds succeed in explosively growing at long times, after a lag time. Finally, for very strong attractions, fractal unbreakable clusters form and grow following DLCA-like (diffusion limited cluster aggregation) kinetics; liquid-gas separation is prevented by the strength of the bonds, which do not allow restructuration. Good qualitative and quantitative agreement is found between the dynamic simulations and the kinetic model in all the three regions. PMID- 17461569 TI - Lateral mobility of polyelectrolyte chains in multilayers. AB - In this work, the lateral mobility of polyelectrolyte multilayers was investigated by means of the fluorescence recovery after photobleaching (FRAP) technique, with special attention to the effect of relevant parameters during and after preparation. Different polyelectrolytes with respect to charge density, stiffness, and hydrophilicity were compared. From the experimental results emerged that the density of charged sites along the polymer is the most important parameter controlling the formation of polymer complexes. At higher charge density, more complexes are formed, and the diffusion coefficient decreases. It was observed that the intrinsic backbone stiffness reduces the interpenetration of polyelectrolyte layers and the formation of complexes promoting the lateral mobility. In addition, the lateral mobility increases with increasing ionic strength and with decreasing hydration shell of the added anion in the polyelectrolyte solution. The effect of heating or annealing in electrolyte solution after preparation was also investigated along with the embedding of the probing layer at controlled distances to the multilayer surface. PMID- 17461570 TI - Highly luminescent zinc(II)-bis(8-hydroxyquinoline) complex nanorods: sonochemical synthesis, characterizations, and protein sensing. AB - Highly luminescent zinc(II)-bis(8-hydroxyquinoline) (Znq(2)) complex nanorods have been synthesized via a sonochemical route from the microemulsion containing zinc acetate and 8-hydroxyquinoline. The scanning electron microscopy (SEM) and transmission electron microscopy (TEM) images showed that the products were rod like morphology with a diameter of about 200-450 nm and a length of about 1-3 microm. A possible mechanism for the formation of Znq(2) nanorods is proposed that the ultrasound wave might urge the collision and fusion of initial Znq(2) nuclei to form nanorods. The photoluminescence (PL) and resonance light scattering (RLS) of the products were also investigated. The Znq(2) nanorods were found to be sensitive to several proteins, such as human serum albumin (HSA), bovine serum albumin (BSA), bovine hemoglobin (Hb), and egg albumin (EA), displaying an increase in intensities of both PL and RLS. The protein concentration dependence of the PL and RLS intensities can be well described as a Langmuir-type binding isotherm. This is the first report on the enhancement of PL and RLS intensities of Znq(2) nanorods by proteins. On the basis of enhanced PL and RLS intensities, the protein could be detected at the nanogram per milliliter level. The experimental results clearly showed that the Znq(2) nanorods were good protein probes for easy and highly sensitive detection. PMID- 17461571 TI - Electrochemistry, spectroscopy, and electrogenerated chemiluminescence of some star-shaped truxene-oligofluorene compounds. AB - We report electrochemical studies, spectroscopy, and electrogenerated chemiluminescence (ECL) of four monodisperse star-shaped truxene core oligofluorene compounds (T1-T4). All oligomers produced stable radical anions and radical cations and showed blue ECL by ion annihilation with an intensity that could be seen with the naked eye. ECL spectra showed that all ECL emissions were at the same position as the fluorescence emission, except for T1, the compound with the shortest fluorene arms that produced some longer wavelength emission in addition to that seen in the fluorescence spectrum. When tetra-n-butylammonium oxalate was used as a coreactant for T1, the emission was much weaker than that in ion annihilation with the same long-wavelength emission observed, making it unlikely that this emission can be ascribed to excimer formation. The ECL intensity of T4 was about 80% of the common blue ECL emitter, 9,10 diphenylanthracene (DPA), under similar conditions. PMID- 17461572 TI - Theoretical study of alpha-84 phycocyanobilin chromophore from the thermophilic cyanobacterium Synechococcus elongatus. AB - Time-dependent density functional theory (TD-DFT) calculations were performed to obtain vertical excitation energies from the ground state to different low-lying singlet excited states of the protonated alpha-84 phycocyanobilin chromophore (alpha-84 PCBH(+)). It clearly emerges that three gradient-corrected approximation functionals (B3LYP, PBE0, and PBEPBE) show a similar description, confirming the proposed valence assignment of the strongest UV-vis absorption band at 618 nm. Moreover, our results show that there are not appreciable differences, in terms of excitation wavelength of the main peak, between the alpha-84 PCBH(+) chromophore and a model system in which the two propionic chains have not been taken into account. Finally, with the precise aim of investigating the effects of alpha-84 PCBH(+) conformational fluctuations on its electronic properties, vertical excitation energies obtained for the potential energy local minimum structure were also refined using a recently proposed TD-DFT/principal component analysis/Car-Parrinello molecular dynamics computational approach. Interestingly, and in line with previous results on another photosensitive complex, this study essentially suggests that interaction with the surrounding environment (protein matrix plus solvent molecules) coupled with the large amplitude fluctuation of the whole C-Phycocyanin (C-PC) pigment protein can affect the electronic properties of the alpha-84 PCB chromophore and therefore its biological activity. PMID- 17461573 TI - Equilibrium states of self-assembly systems: Monte Carlo simulations. AB - We investigated the equilibrium states of the self-assembly of amphiphilic molecules in water. The amphiphiles are represented by chains of the type H1T4, where H is the hydrophilic part of the molecule and T is its hydrophobic portion formed by four monomers. We have performed Monte Carlo simulations on a two dimensional lattice, in which each water molecule occupies a single site, and the amphiphiles occupy five sites of the lattice. We have determined the aggregate distribution curves for the system at low concentration and fixed temperature. We have shown that the criterion to determine the equilibrium states of the system, based on the stabilization of energy curves as a function of the simulation time, is not reliable. The best way to ensure that the equilibrium state was reached was to follow the route to equilibrium of all aggregate sizes of the system. PMID- 17461574 TI - 1D and 2D end-to-end azide-bridged cobalt(II) complexes: syntheses, crystal structures, and magnetic properties. AB - Two new polynuclear azido-bridged Co(II) compounds with formulas catena-[Co(mu1,3 N3)(N3)(py)2(H2O)]n (1) and [Co(mu1,3-N3)2(4-acpy)2]n (2) (py=pyridine, 4-acpy=4 acetylpyridine) have been structurally and magnetically characterized. Compound 1 crystallizes in the orthorhombic system Fddd space group and consists of a single end-to-end azido-bridged chain with the Co(II) atoms in a CoN5O slightly distorted octahedron. Compound 2 crystallizes in the monoclinic system P21/a space group and shows 2D sheets built up through end-to-end azido bridges with the Co(II) atoms in a CoN6 environment. The magnetic properties of 1 and 2 are reported. In the high-temperature region, the plots of chiM or chiMT vs T for 1 and 2 compounds can be fitted by using the Curie-Weiss law, and the best-fit values are -69.1 and -22.6 K, respectively. For 2 magnetic ordering and spontaneous magnetization is achieved below Tc=25 K. PMID- 17461575 TI - Comprehensive molecular mechanics model for oxidized type I copper proteins: active site structures, strain energies, and entatic bulging. AB - The ligand field molecular mechanics (LFMM) model has been applied to the oxidized Type 1 copper center. In conjunction with the AMBER94 force field implemented in DommiMOE, the ligand field extension of the molecular operating environment (MOE), LFMM parameters for Cu-N(imidazole), Cu-S(thiolate), Cu S(thioether), and Cu-O(carbonyl) interactions were developed on the basis of experimental and theoretical data for homoleptic model systems. Subsequent LFMM optimizations of the active site model complex [Cu(imidazole)2(SMe)(SMe2]+ agree with high level quantum results both structurally and energetically. Stable trigonal and tetragonal structures are located with the latter about 1.5 kcal mol 1 lower in energy. Fully optimized unconstrained structures were computed for 24 complete proteins containing T1 centers spanning four-coordinate, plastocyanin like CuN2SS' and stellacyanin-like CuN2SO sites, plus the five-coordinate CuN2SS'O sites of the azurins. The initial structures were based on PDB coordinates augmented by a 10 A layer of water molecules. Agreement between theory and experiment is well within the experimental uncertainties. Moreover, the LFMM results for plastocyanin (Pc), cucumber basic protein (CBP) and azurin (Az) are at least as good as previously reported QM/MM structures and are achieved several orders of magnitude faster. The LFMM calculations suggest the protein provides an entatic strain of about 10 kcal mol-1. However, when combined with the intrinsic 'plasticity' of d9 Cu(II), different starting protein/solvent configurations can have a significant effect on the final optimized structure. This 'entatic bulging' results in relatively large fluctuations in the calculated metal-ligand bond lengths. For example, simply on the basis of 25 different starting configurations of the solvent molecules, the optimized Cu-S(thiolate) bond lengths in Pc vary by 0.04 A while the Cu-S(thioether) distance spans over 0.3 A. These variations are the same order of magnitude as the differences often quoted to correlate the spectroscopic properties from a set of proteins. Isolated optimizations starting from PDB coordinates (or indeed, the PDB structures themselves) may only accidentally correlate with spectroscopic measurements. The present calculations support the work of Warshel who contends that adequate configurational averaging is necessary to make proper contact with experimental properties measured in solution. The LFMM is both sufficiently accurate and fast to make this practical. PMID- 17461576 TI - Solid-state structures and solution studies of novel cyclopentadienyl mercury compounds. AB - New mercury cyclopentadienyl complexes Hg(eta1-Cp')Cl have been prepared by the reaction of HgCl2 and the appropriate KCp' salts or by the transmetalation of HgCl2 with ZnCp'2 (Cp'=C5Me4H, 1; C5Me4But, 2; C5Me4SiMe3, 3; C5H4SiMe3, 4). By contrast, only the SiMe3-substituted bis(cyclopentadienyl) derivatives, Hg(C5Me4SiMe3)2 (5) and Hg(C5H4SiMe3)2 (6), can be isolated by the above synthetic procedures and the appropriate ratio of reagents or from HgCp'Cl and KCp'. Solution NMR studies reveal nonfluxional behavior of the SiMe3-substituted complexes 3, 5, and 6. X-ray studies of the solid-state structures show that the six compounds contain eta1-Cp' ligands, with linear or almost linear C-Hg-Cl or C Hg-C coordination environments. The two HgCp'2 compounds, 5 and 6, have the expected insular structures, but the HgCp'Cl derivatives show supramolecular associations by means of weak secondary Hg...Cl interactions. Thus, the HgCp'Cl compounds 1, 3, and 4 form three different polymeric chain structures with typically two Hg...Cl interactions of 3.04-3.46 A per mercury. By contrast, 2 forms a tetramer, [Hg(C5Me4SiMe3)Cl]4, with a cubelike arrangement of four Hg and four Cl atoms. Density functional theory has been used to investigate the electronic structure of the compounds. PMID- 17461577 TI - Kit for unsymmetric dinucleating double-Schiff-base ligands: facile access to a versatile new ligand system and its first heterobimetallic copper-zinc complex. AB - The synthetic route toward new unsymmetric compartmental "end-off" Schiff-base ligands in a straightforward two-step reaction of 2,6-diformyl-4-methylphenol and two different amine components is presented. To demonstrate the versatility of this method, we have synthesized two different single-Schiff-base proligands, Hbpahmb and Hphmb, utilizing (2-aminoethyl)bis(2-pyridylmethyl)amine and (2 aminomethyl)pyridine, respectively. Subsequent reaction with thiosemicarbazide as the second amine component leads to the novel unsymmetric double-Schiff-base ligands {1-[3-[2-[bis(pyridin-2-ylmethyl)amino]ethyliminomethyl]-2-hydroxy-5 methylphenyl]methylidene}hydrazine carbothioamide (H2bpamptsc) and {1-[3-(pyridin 2-ylmethyliminomethyl)-2-hydroxy-5-methylphenyl]methylidene}hydrazine carbothioamide (H2pmptsc). Both ligands provide two distinctly different coordination pockets: a rigid tridentate N,O,S donor set of the hydrazide compartment versus a rather flexible pentadentate (H2bpamptsc) or tridentate (H2pmptsc) nitrogen-rich chelating side arm. The reaction of the ligand H2bpamptsc with zinc(II) acetate and copper(II) perchlorate yields the heterobinuclear Cu-Zn complex [CuZn(bpamptsc)(mu2,eta1-OAc)(MeCN)](ClO4) (1). PMID- 17461578 TI - Has monopotassium phthalocyanine, KPc, been synthesized? AB - A recent publication described the synthesis and characterization of monopotassium phthalocyanine, KPc. A reevaluation of the original EPR, magnetic susceptibility, and X-ray diffraction data, along with a comparison to the powder diffraction pattern of an authentic sample of Na2Pc, indicates that the material was actually Na2Pc. PMID- 17461579 TI - Influence of chelate substituents on the structure and spin state of unsaturated [N(SiMe2CH2PtBu2)2]Ru-X. AB - Density functional theory calculations on the conformational preferences in the two fused five-membered rings of anionic N(SiR2CH2PR'2)2 chelated to RuX+ are compared to several experimental structures (X=halide). The calculations consider the structures of both singlet and triplet states and reveal that both the four tBu groups and the crowded juncture (N(SiMe2)2) of the two rings must be included computationally to understand the observed structures. Computational experiments with different substituents R and R' show the reality of N-->Ru pi donation. The cases where X=H and CH3 are also studied. PMID- 17461580 TI - Hydrogen-bonded metal-complex sulfonate (MCS) inclusion compounds: effect of the guest molecule on the host framework. AB - Soft molecular host frameworks made of the hydrogen-bonded metal complex (MC) Co(NH3)(6)3+ and 4,4'-biphenyldisulfonate (BPDS) include different guest molecules to form inclusion compounds of the type (MC)2(BPDS)3.n(guest). Structurally characterized were six compounds with guest molecules of DMSO, DMF, piperidine, acetone, acetonitrile, and THF. The metal-complex sulfonate frameworks in all of them are of the pillared layer type where the layers are constructed of extensively hydrogen-bonded metal-complex cations and sulfonate (S) anions (and some hydrogen-bonded water) while the organic residues of the 4,4'-biphenyldisulfonate serve as pillars. The hydrogen-bonded MCS layers and the orientations of the pillars adjust and rearrange in order to generate cavities that would accommodate different guest molecules. The steric, electronic, and hydrogen-bonding needs of the guest molecules mold the soft framework into different structures. These MCS host-guest frameworks are very close structural analogues of the well-studied guanidinium sulfonate (GS) networks and mimic their flexibility and overall durability. PMID- 17461583 TI - Hydroxyquinone O-methylation in mitomycin biosynthesis. AB - Mitomycins are bioreductively activated DNA-alkylating agents. One member of this family, mitomycin C, is in clinical use as part of combination therapy for certain solid tumors. The cytotoxicity displayed by mitomycins is dependent on their electrochemical potential which, in turn, is governed in part by the substituents of the quinone moiety. In this paper we describe studies on the biogenesis of the quinone methoxy group present in mitomycins A and B. An engineered Streptomyces lavendulae strain in which the mmcR methyltransferase gene had been deleted failed to produce the three mitomycins (A, B, and C) that are typically isolated from the wild type organism. Analysis of the culture extracts from the mmcR-deletion mutant strain revealed that two new metabolites, 7-demethylmitomycin A and 7-demethylmitomycin B, had accumulated instead. Production of mitomycins A and C or mitomycin B was selectively restored upon supplementing the culture medium of a S. lavendulae strain unable to produce the key precursor 3-amino-5-hydroxybenzoate with either 7-demethylmitomycin A or 7 demethylmitomycin B, respectively. MmcR methyltransferase obtained by cloning and overexpression of the corresponding mmcR gene was shown to catalyze the 7-O methylation of both C9beta- and C9alpha-configured 7-hydroxymitomycins in vitro. This study provides direct evidence for the catalytic role of MmcR in formation of the 7-OMe group that is characteristic of mitomycins A and B and demonstrates the prerequisite of 7-O-methylation for the production of the clinical agent mitomycin C. PMID- 17461584 TI - Acetylcholinesterase: mechanisms of covalent inhibition of wild-type and H447I mutant determined by computational analyses. AB - The reaction mechanisms of two inhibitors TFK+ and TFK0 binding to both the wild type and H447I mutant mouse acetylcholinesterase (mAChE) have been investigated by using a combined ab initio quantum mechanical/molecular mechanical (QM/MM) approach and classical molecular dynamics (MD) simulations. In the wild-type mAChE, the binding reactions of TFK+ and TFK0 are both spontaneous processes, which proceed through the nucleophilic addition of the Ser203-Ogamma to the carbonyl-C of TFK+ or TFK0, accompanied with a simultaneous proton transfer from Ser203 to His447. No barrier is found along the reaction paths, consistent with the experimental reaction rates approaching the diffusion-controlled limit. By contrast, TFK+ binding to the H447I mutant may proceed with a different reaction mechanism. A water molecule takes over the role of His447 and participates in the bond breaking and forming as a "charge relayer". Unlike in the wild-type mAChE case, Glu334, a conserved residue from the catalytic triad, acts as a catalytic base in the reaction. The calculated energy barrier for this reaction is about 8 kcal/mol. These predictions await experimental verification. In the case of the neutral ligand TFK0, however, multiple MD simulations on the TFK0/H447I complex reveal that none of the water molecules can be retained in the active site as a "catalytic" water. Furthermore, our alchemical free energy calculation also suggests that the binding of TFK0 to H447I is much weaker than that of TFK+. Taken together, our computational studies confirm that TFK0 is almost inactive in the H447I mutant and also provide detailed mechanistic insights into the experimental observations. PMID- 17461585 TI - Proton-abstraction mechanism in the palladium-catalyzed intramolecular arylation: substituent effects. AB - The regioselectivity observed in the intramolecular palladium-catalyzed arylation of substituted bromobenzyldiarylmethanes as well as theoretical results demonstrate that the Pd-catalyzed arylation proceeds by a mechanism involving a proton abstraction by the carbonate, or a related basic ligand. The reaction is facilitated by electron-withdrawing substituents on the aromatic ring, which is inconsistent with an electrophilic aromatic-substitution mechanism. The more important directing effect is exerted by electron-withdrawing substituents ortho to the reacting site. PMID- 17461586 TI - Glutathione- and cysteine-induced transverse overgrowth on gold nanorods. PMID- 17461587 TI - Resonance Raman detection of the hydroperoxo intermediate in the cytochrome P450 enzymatic cycle. PMID- 17461588 TI - Reactivity of platinum metal with organic radical anions from metal to negative oxidation states. AB - The reaction of platinum metal with an organic molecular radical anion leads to the formation of iono-metallic phases where Pt exists under negative oxidation states. This puzzling transformation of a "noncorrodible metal" was examined using localized electrochemical techniques in dimethylformamide containing different tetra-alkylammonium salts chosen as test systems. Our experiments demonstrate that the platinum metal is locally reduced as soon as the Pt faces relatively moderate reducing conditions, for example, when the Pt is used as a negative electrode or when the metal is in the presence of a reducing agent such as an organic radical anion. Scanning electrochemical microscopy (SECM) analysis, current-distance curves, and transient mode responses provide detailed descriptions of the reactivity of Pt to form negative oxidation states (the key step is the reaction of the metal with a molecular reducing agent), of the insulating nature of the "reduced" solid phases of the thermodynamics and kinetics conditions of the Pt conversion. The passage from the conductor to insulator states controlled the spatial development of the reaction that always remains in competition with the other "natural" roles of a metallic electrode. Formally, the phenomena can be treated by analogy with the C. Amatore's model previously developed for the mediated reduction of the poly(tetrafluoroethylene). Consequences of this general reactivity of Pt are discussed in view of a wide utilization of this metal in reductive conditions and the possible applications of such processes in the micropatterning of metallic surfaces. PMID- 17461589 TI - Cinchona alkaloid-catalyzed enantioselective monofluoromethylation reaction based on fluorobis(phenylsulfonyl)methane chemistry combined with a Mannich-type reaction. PMID- 17461590 TI - Thermal and photochemical switching of conformation of poly(ethylene glycol) substituted cyclodextrin with an azobenzene group at the chain end. PMID- 17461592 TI - In situ measurements of pH changes in beta-lactoglobulin solutions under high hydrostatic pressure. AB - A novel in situ method, in which the spectral changes of aqueous solutions under pressure are measured using optical pH indicators in a high-pressure spectrophotometer, has been developed in order to provide a quantitative description of the pressure dependence of acid/base equilibria of proteins. The self-consistent method, insensitive to compressibility, was developed for measurement of changes in pH with pressure based on alpha-naphthyl red and neutral red as these indicators were found to have pressure insensitive pKa values. The method was validated for up to 500 MPa by measurement of the pressure dependence of the weak acid buffers acetic acid/acetate and imidazolium/imidazole from which volumes of dissociation of DeltaV degrees = -11.2 and 3.7 mL/mol, respectively, were established. Succinic acid/hydrogensuccinate was surprisingly insensitive to pressure with DeltaV degrees = -0.9 mL/mol. For beta-lactoglobulin B in an unbuffered aqueous solution with ionic strength of 0.05 M and pH 4, pressure up to 300 MPa increased pH up to 1.5 units depending on concentration (up to 5 mg/mL investigated), followed by a decrease to the initial pH 4 for pressure up to 500 MPa. The surprising increase in pH at pressure up to 300 MPa is suggested to be caused by an increase in the effective pKa values of aspartic acid and glutamic acid side chain in hydrophobic compartments of the protein created by pressure denaturation, leading to a binding of water protons and an increase in free hydroxide ions. For higher pressure the carboxylic side chains in the fully denatured protein again becomes exposed to the solvent, and pH decreases to the initial pH of the aqueous system. PMID- 17461593 TI - Evaluation of occasional nonresponse of a washed cod mince model to hemoglobin (Hb)-mediated oxidation. AB - An emerging model to test antioxidants for application in seafoods is washed cod mince fortified with hemoglobin (Hb) as a catalyst. This system has been used to test the antioxidative activity of certain muscle extracts and some pure compounds such as BHA, BHT, TBHQ, and propyl gallate during ice storage. However, the washed cod mince model has occasionally been resistant to Hb-mediated oxidation. This has been in cases when the moisture of the model has been minimized by washes at the protein isoelectric point (pH approximately 5.5) to allow for large additions of potentially antioxidative solutions. In this paper, noncontrollable and controllable factors for this intriguing occasional oxidation resistance were studied. Compositional analyses (lipid content, alpha-tocopherol, and lipid hydroperoxides) and structural analysis of a "normal" oxidizing model and a stable model were done to identify any differences among them. Some controllable factors related to the model preparation that were studied included different washing pH values (5.5-6.6), Hb concentrations (7.2 and 13.5 microM), final model moisture contents (75, 81, and 90%), and light exposure during ice storage (0 h, 3-4 h, or 24 h of light/day). Results revealed a 2-fold higher alpha-tocopherol content in the stable model than in the oxidizing model. Electron microscopy images showed a more and less disrupted myofibrillar structure in the stable and the oxidizing cod model, respectively. This indicated that "cold setting" (i.e., pre-gelation) of the stable model may have occurred and prevented Hb from diffusing freely in the model. Controllable factors that reduced lipid oxidation in the models were less Hb and lower moisture. PMID- 17461594 TI - Grape-seed procyanidins act as antiinflammatory agents in endotoxin-stimulated RAW 264.7 macrophages by inhibiting NFkB signaling pathway. AB - Procyanindin extract (PE) is a mixture of polyphenols, mainly procyanidins, obtained from grape seed with putative antiinflammatory activity. We evaluated the PE effect on RAW 264.7 macrophages stimulated with lipopolysaccharide plus interferon-gamma that show a rapid enhanced production of prostaglandin E2 (PGE2) and nitric oxide (NO). Our results demonstrated that PE significantly inhibited the overproduction of NO, dose and time dependently. PE caused a marked inhibition of PGE2 synthesis when administered during activation. Moreover, PE pretreatment diminished iNOS mRNA and protein amount dose dependently (10-65 microg/mL). PE (65 microg/mL) pretreatment inhibited NFkappaB (p65) translocation to nucleus by nearly 40%. Trimeric and longer oligomeric-rich procyanidin fractions from PE (5-30 microg/mL) inhibited iNOS expression but not the monomeric forms catechin and epicatechin. Thus, we show that the degree of polymerization is important in determining procyanidin effects. PE was considerably a more effective inhibitor of NO biosynthesis (IC50 = 50 microg/mL) in comparison to other antiinflammatories, such as aspirin (3 mM), indomethacin (20 microM), and dexamethasone (9 nM). In conclusion, PE modulates inflammatory response in activated macrophages by the inhibition of NO and PGE2 production, suppression of iNOS expression, and NFkB translocation. These results demonstrate an immunomodulatory role of grape seed procyanidins and thus a potential health benefit in inflammatory conditions that exert an overproduction of NO and PGE2. PMID- 17461595 TI - Evaluation of the phytoestrogenic activity of Cyclopia genistoides (honeybush) methanol extracts and relevant polyphenols. AB - Unfermented C. genistoides methanol extracts of different harvestings and selected polyphenols were evaluated for phytoestrogenic activity by comparing binding to both ER subtypes, transactivation of an ERE-containing promoter reporter, proliferation of MCF-7-BUS and MDA-MB-231 breast cancer cells, and binding to SHBG. The extracts from one harvesting of C. genistoides (P104) bound to both ER subtypes. All extracts transactivated ERE-containing promoter reporters via ERbeta but not via ERalpha. All extracts, except P122, caused proliferation of the estrogen-sensitive MCF-7-BUS cells. Proliferation of MCF-7 BUS cells was ER-dependent as ICI 182,780 reversed proliferation. Physiologically more relevant, extracts antagonized E2-induced MCF-7-BUS cell proliferation. Furthermore, all extracts, except P122, induced proliferation of the estrogen insensitive MDA-MB-231 cells, suggesting that the extracts are able to induce ER dependent and ER-independent cell proliferation. Binding to SHBG by extracts was also demonstrated. These results clearly show that C. genistoides methanol extracts display phytoestrogenic activity and act predominantly via ERbeta. HPLC and LC-MS analysis, however, suggests that the observed phytoestrogenic activity cannot be ascribed to polyphenols known to be present in other Cyclopia species. PMID- 17461596 TI - Residue depletion of ractopamine and its metabolites in swine tissues, urine, and serum. AB - Ractopamine hydrochloride is a beta-adrenergic leanness-enhancing agent approved for use in swine in the United States. Depletion of ractopamine and its metabolites from animal tissues, urine, and serum is of interest for the detection of illegal use. The objectives of this study were to measure the residues of ractopamine in swine incurred samples after treatment with dietary ractopamine for 28 consecutive days. An efficient and sensitive analytical method was developed for the detection of parent ractopamine and its metabolites in swine tissues, urine, and serum by HPLC-FLD. After extraction, enzymatic digestion, and solid-phase cleanup of the samples, ractopamine residues were determined by liquid chromatography (LC) with fluorescence detector. The limits of detection (LOD) for tissues, urine, and serum were 1 ng g(-1), 0.5 ng mL(-1), and 0.5 ng mL(-1), respectively. Recoveries ranged from 70.5 to 94.5% for samples fortified at 1-50 ng g(-1) or ng mL(-1). Sixty pigs were fed twice daily for 28 consecutive days with feeds containing 18 mg kg(-1) ractopamine HCl. The residue concentrations in urine, liver, and kidney were 650.06 ng mL(-1), 46.09 ng g(-1), and 169.27 ng g(-1), respectively, compared with those in muscle, fat, and serum (4.94 ng g(-1), 3.28 ng g(-1), and 7.48 ng mL(-1), respectively) at the feeding period of 7 days. The residue concentrations at withdrawal period of 0 days in all edible tissues were lower than tolerance values established by the FDA and MRL values listed by the JECFA. These data support the withdrawal time of 0 days established by the FDA for ractopamine used as feed additive in swine. PMID- 17461597 TI - The salt-cocrystal continuum: the influence of crystal structure on ionization state. AB - Salts and cocrystals are multicomponent crystals that can be distinguished by the location of the proton between an acid and a base. At the salt end of the spectrum proton transfer is complete, and on the opposite end proton transfer is absent in cocrystals. However, for acid-base complexes with similar pK(a) values, the extent of proton transfer in the solid state is not predictable and a continuum exists between the two extremes. For these systems, both the DeltapK(a) value (pK(a) of base - pK(a) of acid) and the crystalline environment determine the extent of proton transfer. A total of 20 complexes containing theophylline and guest molecules with DeltapK(a) values less than 3 have been prepared, resulting in 13 cocrystals, five salts, and two complexes with mixed ionization states based on IR spectroscopy and single-crystal diffraction data. We propose modifications to the DeltapK(a) rule for selecting salt screen counterions that focus on the discovery of solid forms with useful physical properties rather than an arbitrary cutoff value for DeltapK(a). PMID- 17461598 TI - Dipyrromethane + dipyrromethanedicarbinol routes to an electron deficient meso substituted phlorin with enhanced stability. AB - Two dipyrromethane + dipyrromethanedicarbinol routes to a meso-substituted phlorin bearing electron-withdrawing pentafluorophenyl substituents (TpFPPhl) were investigated in an attempt to obtain a phlorin with enhanced stability toward light and air and to explore the application of dipyrromethanecarbinol chemistry to the preparation of phlorins. For each route, a systematic survey of reaction parameters for the two-step, one-flask reaction leading to TpFPPhl was performed. The analytical-scale reactions were monitored for yield of TpFPPhl by HPLC. Sharp differences were observed in the yield of TpFPPhl afforded by the two synthetic routes. The most promising reaction condition (TFA catalysis, 100 mM) was performed on a preparative scale providing TpFPPhl in a yield of 45% (189 mg). The stability of the electron-deficient phlorin in dilute solution upon exposure to light and air was probed in a number of solvents, and decomposition was monitored by UV-vis spectroscopy and HPLC. Many of the solutions of TpFPPhl were found to be quite stable for periods of approximately 8 h, with decomposition requiring exposure periods of several days. Taken together, this work contributes an efficient synthesis of a meso-substituted phlorin of practical stability and provides further insights toward the adaptation of dipyrromethanecarbinol chemistry to the preparation of diverse porphyrinoids. PMID- 17461599 TI - Glycosides from the stem bark of Fraxinus sieboldiana. AB - A norditerpene glucopyranoside with a novel carbon skeleton (1), eight new aromatic glycosides (2-9), and 25 known glycosides have been isolated from a H2O soluble portion of an ethanolic extract of the stem bark of Fraxinus sieboldiana. Their structures were determined by spectroscopic and chemical methods. Based on analysis of the NMR data of threo- and erythro-arylglycerols in different solvents, an application of Delta delta C8-C7 values to distinguish threo arylglycerol and erythro-arylglycerol isomers was proposed. In the in vitro assays, compound 5 displayed TNF-alpha secretion inhibitory activity with an IC50 value of 1.6 microM, compound 6 showed antioxidative activity inhibiting Fe+2 cystine-induced rat liver microsomal lipid peroxidation with an IC50 value of 0.9 microM, and plantasioside (10) showed selective activity against the human colon cancer cell line (HCT-8) with an IC50 value of 3.4 microM. PMID- 17461600 TI - In situ auger electron spectroscopy study of atomic layer deposition: growth initiation and interface formation reactions during ruthenium ALD on Si-H, SiO2, and HfO2 surfaces. AB - Growth initiation and film nucleation in atomic layer deposition (ALD) is important for controlling interface composition and achieving atomic-scale films with well-defined composition. Ruthenium ALD is studied here using ruthenocene and oxygen as reactants, and growth initiation and nucleation are characterized on several different growth surfaces, including SiO2, HfO2, and hydrogen terminated silicon, using on-line Auger electron spectroscopy and ex-situ X-ray photoelectron spectroscopy. The time needed to reach the full growth rate (typically approximately 1 A per deposition cycle) is found to increase as the surface energy of the starting surface (determined from contact angle measurements) decreased. Growth starts more readily on HfO2 than on SiO2 or Si-H surfaces, and Auger analysis indicates distinct differences in surface reactions on the various surfaces during film nucleation. Specifically, surface oxygen is consumed during ruthenocene exposure, so the nucleation rate will depend on the availability of oxygen and the energetics of surface oxygen bonding on the starting substrate surface. PMID- 17461601 TI - Actin network formation by unidirectional polycation diffusion. AB - We show that F-actins form three-dimensional giant network under uni-directional diffusion of polycations, at a dilute actin concentration (0.01 mg/mL) that only bundles are formed by homogeneous mixing with polycations. The mesh size of the actin network depends on polycation concentration and ionic strength, while bundle thickness of network depends only on ionic strength, which indicates that actin network is formed through nucleation-growth mechanism. The mesh size and the bundle thickness are determined by nucleus concentration and nucleus size, respectively. The atomic force microscopy measurement correlates the elasticity of the actin network, E, with the mesh size, xi, as E approximately xi-1, while the bundle thickness, D dependence of E cannot be described by a simple scaling relation. E approximately D6.5 when D is small and E approximately D0.1 when D is large. Our study on the self-assembly of actin network under asymmetric polycation condition would provide the crucial insight into the organization of biopolymers in polarized condition of cell. PMID- 17461602 TI - Core-shell magnetite nanoparticles surface encapsulated with smart stimuli responsive polymer: synthesis, characterization, and LCST of viable drug targeting delivery system. AB - We describe here the synthesis of a novel magnetic drug-targeting carrier characterized by a core-shell structure. The core-shell carrier combines the advantages of a magnetic core and the stimuli-responsive property of the thermosensitive biodegradable polymer shell (e.g., an on-off mechanism responsive to external temperature change). The composite nanoparticles are approximately 8 nm in diameter with approximately 3 nm shell. The lower critical solution temperature (LCST) is approximately 38 degrees C as determined by UV-vis absorption spectroscopy. The carrier is composed of cross-linked dextran grafted with a poly(N-isopropylacrylamide-co-N,N-dimethylacrylamide) [dextran-g poly(NIPAAm-co-DMAAm)] shell and superparamagnetic Fe3O4 core. Fourier transform infrared spectroscopy (FTIR) confirmed the composition of the carrier. The synthesized magnetic carrier system has potential applications in magnetic drug targeting delivery and magnetic resonance imaging. PMID- 17461603 TI - Cytochrome c self-assembly on alkanethiol monolayer electrodes as characterized by AFM, IR, QCM, and direct electrochemistry. AB - With the advantage of carbodiimide coupling chemistry, horse heart cytochrome c (cyt c) has been covalently immobilized onto self-assembled monolayers (SAMs) from 11-mercaptoundecanoic acid (MUDA) developed on single-crystal or polycrystalline gold substrate surfaces. The cyt c immobilized substrates thus prepared have been characterized by atomic force microscopy (AFM); we have succeeded in obtaining surface topographical images down to single-protein resolution. AFM imaging has also shown densely packed, uniform protein monolayer formation that is highly suggestive of self-assembly of cyt c molecules on MUDA SAMs. Covalent attachment of cyt c has been further evidenced by reflection absorption FT-IR as well as microgravimetric analysis using a quartz crystal microbalance (QCM). In the latter, the specific MUDA and cyt c surface concentrations were determined to be 0.86 +/- 0.11 nmol cm-2 (n = 5) and 28 +/- 12 pmol cm-2 (n = 5), both of which agree fairly well with their theoretical counterparts. The obtained QCM chips having the cyt c/MUDA/Au interfacial structure were found to be capable of the direct electrochemistry of the surface attached cyt c molecules. Cyclic voltammetric measurements on the chips gave particular redox waves showing characteristics of surface process. The electroactive protein surface concentration was determined to be 7.2 +/- 4.8 pmol cm-2 (n = 6); it was almost consistent with values found in literature, while it was limited to 26% in magnitude for the QCM data. This was deemed to have arisen from the orientation variation of the surface-confined cyt c molecules and is discussed briefly. PMID- 17461604 TI - Entropic stabilization and equilibrium size of lipid vesicles. AB - We have studied the phase behavior of zwitterionic phospholipid dioleoylphosphatidylcholine (DOPC) vesicles (membranes) and interpreted our results using scaling arguments in combination with molecular realistic self consistent field (SCF) calculations. DOPC membranes acquire a partial negative charge per lipid molecule at intermediate NaBr concentrations. As a result of this, dilute DOPC solutions form stable unilamellar vesicles. Both at low and high salt concentrations phase separation into a lamellar and a vesicular phase is observed. The vesicle radius decreases as a power law with decreasing lipid concentration. This power-law concentration dependence indicates that the vesicle phase is entropically stabilized; the size of the DOPC vesicles result from a competition between the bending energy and translation and undulation entropy. This scaling behavior breaks down for very small vesicles. This appears to be consistent with SCF predictions that point to the fact that in this regime the mean bending modulus kc increases with curvature. The SCF theory predicts that, at low ionic strength, the membrane stability improves when there is more charge on the lipids. Upon a decrease of the ionic strength, lipids with a full negative charge form vesicles that grow exponentially in size because the mean bending modulus increases with decreasing ionic strength. At the same time the Gaussian bending modulus becomes increasingly negative such that the overall bending energy tends to zero. This indicates that small micelles become the dominant species. The SCF theory thus predicts a catastrophic break down of giant vesicles in favor of small micelles at sufficiently low ionic strength and high charge density on the lipids. PMID- 17461605 TI - Intrinsic current-voltage characteristics of graphene nanoribbon transistors and effect of edge doping. AB - We demonstrate that the electronic devices built on patterned graphene nanoribbons (GNRs) can be made with atomic-perfect-interface junctions and controlled doping via manipulation of edge terminations. Using first-principles transport calculations, we show that the GNR field effect transistors can achieve high performance levels similar to those made from single-walled carbon nanotubes, with ON/OFF ratios on the order of 10(3)-10(4), subthreshold swing of 60 meV per decade, and transconductance of 9.5 x 10(3) Sm-1. PMID- 17461606 TI - Optical properties of a wrinkled nanomembrane with embedded quantum well. AB - A wrinkled nanomembrane with embedded quantum well (QW), fabricated by the partial release and bond back of epitaxial layers upon underetching, is investigated by spatially resolved micro-photoluminescence spectroscopy. From the observed QW transition energies and calculations based on the linear deformation potential theory, we find that the bonded back regions are fully relaxed and act on the strain state of the wrinkled QW. Light emission enhancement observed in the wrinkled QW is explained by interference contrast theory. PMID- 17461607 TI - Interaction of an infrared surface plasmon with an excited molecular vibration. AB - The interaction of an infrared surface plasmon and an excited molecular vibration was investigated by using a square array of subwavelength holes in a Ni film which supports propagating, surface-plasmon-mediated, transmission resonances. The largest transmission resonance [the (1,0)(-)] was tuned through the rocking vibration of the hexadecane molecule (at 721 cm(-1)) in a hexadecane film on the mesh by varying the thickness of the film. The interaction of the rocking vibration and surface plasmon is characterized spectroscopically by an increase in the intensity of the vibrational band by more than a factor of 2, variation of the vibrational line shape relative to the spectrum on a nonmetallic surface, and shifts in vibrational peak position by as much as 3.0 cm(-1). Relationships are developed between the transmission resonance position and the thickness and dielectric properties of the coating. PMID- 17461608 TI - Frequency-dependent nonlinear optical properties with explicitly correlated coupled-cluster response theory using the CCSD(R12) model. AB - Response theory up to infinite order is combined with the explicitly correlated coupled-cluster singles and doubles model including linear-r(12) corrections, CCSD(R12). The additional terms introduced by the linear-r(12) contributions, not present in the conventional CCSD calculation, are derived and discussed with respect to the extra costs required for their evaluation. An implementation is presented up to the cubic response function for one-electron perturbations, i.e., up to frequency-dependent second hyperpolarizabilities. As first applications the authors computed the electronic polarizabilities and second hyperpolarizabilities of BH, N(2), and formaldehyde and show that the improvement in the one-electron basis set convergence known from the R12 method for ground state energies is retained for higher-order optical properties. Frequency-dependent results are presented for the second hyperpolarizability of N(2). PMID- 17461609 TI - Chemical origin of blue- and redshifted hydrogen bonds: intramolecular hyperconjugation and its coupling with intermolecular hyperconjugation. AB - Upon formation of a H bond Y...H-XZ, intramolecular hyperconjugation n(Z)- >sigma*(X-H) of the proton donor plays a key role in red- and blueshift characters of H bonds and must be introduced in the concepts of hyperconjugation and rehybridization. Intermolecular hyperconjugation transfers electron density from Y to sigma*(X-H) and causes elongation and stretch frequency redshift of the X-H bond; intramolecular hyperconjugation couples with intermolecular hyperconjugation and can adjust electron density in sigma*(X-H); rehybridization causes contraction and stretch frequency blueshift of the X-H bond on complexation. The three factors--intra- and intermolecular hyperconjugations and rehybridization--determine commonly red- or blueshift of the formed H bond. A proton donor that has strong intramolecular hyperconjugation often forms blueshifted H bonds. PMID- 17461610 TI - Calculation of the electric hypershielding at the nuclei of molecules in a strong magnetic field. AB - The third-rank electric hypershielding at the nuclei of 14 small molecules has been evaluated at the Hartree-Fock level of accuracy, by a pointwise procedure for the geometrical derivatives of magnetic susceptibilities and by a straightforward use of its definition within the Rayleigh-Schrodinger perturbation theory. The connection between these two quantities is provided by the Hellmann-Feynman theorem. The magnetically induced hypershielding at the nuclei accounts for distortion of molecular geometry caused by strong magnetic fields and for related changes of magnetic susceptibility. In homonuclear diatomics H(2), N(2), and F(2), a field along the bond direction squeezes the electron cloud toward the center, determining shorter but stronger bond. It is shown that constraints for rotational and translational invariances and hypervirial theorems provide a natural criterion for Hartree-Fock quality of computed nuclear electric hypershielding. PMID- 17461611 TI - Computation of correlation functions and wave function projections in the context of quantum trajectory dynamics. AB - The de Broglie-Bohm formulation of the Schrodinger equation implies conservation of the wave function probability density associated with each quantum trajectory in closed systems. This conservation property greatly simplifies numerical implementations of the quantum trajectory dynamics and increases its accuracy. The reconstruction of a wave function, however, becomes expensive or inaccurate as it requires fitting or interpolation procedures. In this paper we present a method of computing wave packet correlation functions and wave function projections, which typically contain all the desired information about dynamics, without the full knowledge of the wave function by making quadratic expansions of the wave function phase and amplitude near each trajectory similar to expansions used in semiclassical methods. Computation of the quantities of interest in this procedure is linear with respect to the number of trajectories. The introduced approximations are consistent with approximate quantum potential dynamics method. The projection technique is applied to model chemical systems and to the H+H(2) exchange reaction in three dimensions. PMID- 17461612 TI - Long-range corrected density functional calculations of chemical reactions: redetermination of parameter. AB - Chemical reaction calculations were carried out using the long-range correction (LC) scheme, which improves long-range exchange effects in density functional theory (DFT) [J. Chem. Phys. 115, 3540 (2001); 120, 8425 (2004)]. A new determination of the LC scheme parameter mu was made by a root mean square fit of the percent error in calculated atomization energies. As a result, the parameter mu was optimized as 0.47, which is higher than the previous one (mu=0.33). Using this new parameter mu, LC-DFT was firstly applied to geometry optimizations of the G2 benchmark set molecules. Consequently, this new LC-DFT gave more accurate bond lengths and bond angles than previous LC-DFT and hybrid B3LYP results. Following this result, the authors calculated reaction barrier height energies of benchmark reaction sets, which have been underestimated in conventional DFT calculations. Calculated results showed that LC-DFT provided much more accurate barrier height energies with errors less than half those of previous LC-DFT and B3LYP studies. To test the general validity of the new LC-DFT, the authors finally calculated reaction enthalpies. As a result, they found that the LC scheme using the new mu clearly improved the accuracy of calculated enthalpies. The authors therefore conclude that the insufficient inclusion of long-range exchange effects is responsible for the underestimation of reaction barriers in DFT calculations and that LC-DFT using the new parameter is a powerful tool for theoretically investigating chemical reactions. PMID- 17461613 TI - Relativistic two-component infinite order method for atomic core ionization potentials. AB - In this paper the authors have applied the infinite-order two-component method (IOTC) to compute the valence and inner shell ionization potentials for the Ne, Ar, Kr, and Xe elements. The obtained results show the very good performance of the recently defined relativistic IOTC method. They also confirm the importance of the relativistic effects in the determination of the inner shell ionization potentials. PMID- 17461614 TI - Decay rates of inner-valence excitations in noble gas atoms. AB - A Fano - algebraic diagrammatic construction - Stieltjes method has been recently developed for ab initio calculations of nonradiative decay rates [V. Averbukh and L. S. Cederbaum, J. Chem. Phys. 123, 204107 (2005)] of singly ionized states. In the present work this method is generalized for the case of electronic decay of excited states. The decay widths of autoionizing inner-valence-excited states of Ne, Ar, and Kr are calculated. Apart from the lowest excitation of Kr, they are found to be in good to excellent agreement with the experimental values. Comparison with the other theoretical studies shows that in many cases the new method performs better than the previously available techniques. PMID- 17461615 TI - Linear-scaling implementation of molecular response theory in self-consistent field electronic-structure theory. AB - A linear-scaling implementation of Hartree-Fock and Kohn-Sham self-consistent field theories for the calculation of frequency-dependent molecular response properties and excitation energies is presented, based on a nonredundant exponential parametrization of the one-electron density matrix in the atomic orbital basis, avoiding the use of canonical orbitals. The response equations are solved iteratively, by an atomic-orbital subspace method equivalent to that of molecular-orbital theory. Important features of the subspace method are the use of paired trial vectors (to preserve the algebraic structure of the response equations), a nondiagonal preconditioner (for rapid convergence), and the generation of good initial guesses (for robust solution). As a result, the performance of the iterative method is the same as in canonical molecular-orbital theory, with five to ten iterations needed for convergence. As in traditional direct Hartree-Fock and Kohn-Sham theories, the calculations are dominated by the construction of the effective Fock/Kohn-Sham matrix, once in each iteration. Linear complexity is achieved by using sparse-matrix algebra, as illustrated in calculations of excitation energies and frequency-dependent polarizabilities of polyalanine peptides containing up to 1400 atoms. PMID- 17461616 TI - Tests of functionals for systems with fractional electron number. AB - In the exact theory, the ground state energy of an open system varies linearly when the electron number is changed between two adjacent integers. This linear dependence is not reproduced by common approximate density functionals. Deviation from linearity in this dependence has been suggested as a basis for the concept of many-electron self-interaction error (SIE). In this paper, we quantify many electron SIE of a number of approximations by performing calculations on fractionally charged atoms. We demonstrate the direct relevance of these studies to such problems of common approximate functionals as instabilities of anions, spurious fractional charges on dissociated atoms, and poor description of charge transfer. Semilocal approximations have the largest many-electron SIE, which is only slightly reduced in typical global hybrids. In these approximations the energy versus fractional electron number curves upward, while in Hartree-Fock theory the energy curves downward. Perdew-Zunger self-interaction correction [Phys. Rev. B 23, 5048 (1981)] significantly reduces the many-electron SIE of semilocal functionals but impairs their accuracy for equilibrium properties. In contrast, a long-range corrected hybrid functional can be nearly many-electron SIE-free in many cases (for reasons we discuss) and at the same time performs remarkably well for many molecular properties. PMID- 17461617 TI - Fully automated implementation of the incremental scheme: application to CCSD energies for hydrocarbons and transition metal compounds. AB - A general fully automated implementation of the incremental scheme for molecules and embedded clusters in the framework of the coupled cluster singles and doubles theory is presented. The code can be applied to arbitrary order of the incremental expansion and is parallelized in a master/slave structure. The authors found that the error in the total correlation energy is lower than 1 kcal/mol with respect to the canonical CCSD calculation if the incremental series is truncated in a proper way. PMID- 17461618 TI - Gauge invariant calculations of nuclear magnetic shielding constants using the continuous transformation of the origin of the current density approach. II. Density functional and coupled cluster theory. AB - The quantum mechanical current density induced in a molecule by an external magnetic field is invariant to translations of the coordinate system. This fundamental symmetry is exploited to formally annihilate the diamagnetic contribution to the current density via the approach of "continuous transformation of the origin of the current density-diamagnetic zero" (CTOCD-DZ). The relationships obtained by this method for the magnetic shielding at the nuclei are intrinsically independent of the origin of the coordinate system for any approximate computational scheme relying on the algebraic approximation. The authors report for the first time an extended series of origin-independent estimates of nuclear magnetic shielding constants using the CTOCD-DZ approach at the level of density functional theory (DFT) with four different types of functionals and unrelaxed coupled cluster singles and doubles linear response (CCSD-LR) theory. The results obtained indicate that in the case of DFT the procedure employed is competitive with currently adopted computational methods allowing for basis sets of gauge-including atomic orbitals, whereas larger differences between CTOCD-DZ and common origin CCSD-LR results are observed due to the incomplete fulfillment of hypervirial relations in standard CCSD-LR theory. It was found furthermore that the unrelaxed CCSD-LR calculations predict larger correlation corrections for the shielding constants of almost all nonhydrogen atoms in their set of molecules than the usual relaxed energy derivative CCSD calculations. Finally the results confirm the excellent performance of Keal and Tozer's third functional, in particular, for the multiply bonded systems with a lot of electron correlation, but find also that the simple local density functional gives even better results for the few singly bonded molecules in their study where correlation effects are small. PMID- 17461619 TI - Density functional self-consistent quantum mechanics/molecular mechanics theory for linear and nonlinear molecular properties: Applications to solvated water and formaldehyde. AB - A combined quantum mechanics/molecular mechanics (QM/MM) method is described, where the polarization between the solvent and solute is accounted for using a self-consistent scheme linear in the solvent polarization. The QM/MM method is implemented for calculation of energies and molecular response properties including the calculation of linear and quadratic response functions using the density-functional theory (DFT) and the Hartree-Fock (HF) theory. Sample calculations presented for ground-state energies, first-order ground-state properties, excitation energies, first-order excited state properties, polarizabilities, first-hyperpolarizabilities, and two-photon absorptions strengths of formaldehyde suggests that DFT may in some cases be a sufficiently reliable alternative to high-level theory, such as coupled-cluster (CC) theory, in modeling solvent shifts, whereas results obtained with the HF wave function deviate significantly from the CC results. Calculations carried out on water gives results that also are comparable with CC calculations in accuracy for ground-state and first-order properties. However, to obtain such accuracy an exchange-correlation functional capable of describing the diffuse Rydberg states must be chosen. PMID- 17461620 TI - Photodissociation of CO2 - in water clusters via Renner-Teller and conical interactions. AB - The photochemistry of mass selected CO(2) (-)(H2O)(m), m=2-40 cluster anions is investigated using 266 nm photofragment spectroscopy and theoretical calculations. Similar to the previous 355 nm experiment [Habteyes et al., Chem. Phys. Lett. 424, 268 (2006)], the fragmentation at 266 nm yields two types of anionic products: O(-)(H2O)(m-k) (core-dissociation products) and CO(2) ( )(H2O)(m-k) (solvent-evaporation products). Despite the same product types, different electronic transitions and dissociation mechanisms are implicated at 355 and 266 nm. The 355 nm dissociation is initiated by excitation to the first excited electronic state of the CO(2) (-) cluster core, the 1 (2)B(1)(2A") state, and proceeds via a glancing Renner-Teller intersection with the ground electronic state at a linear geometry. The 266 nm dissociation involves the second excited electronic state of CO(2) (-), the 2 (2)A(1)(2A') state, which exhibits a conical intersection with the 3 (2)B(2)(A') state at a bent geometry. The asymptotic O(-) based products are believed to be formed via this 3 (2)B(2)(A') state. By analyzing the fragmentation results, the bond dissociation energy of CO(2) (-) to O(-)+CO in hydrated clusters (m> or =20) is estimated as 2.49 eV, compared to 3.46 eV for bare CO(2) (-). The enthalpy of evaporation of one water molecule from asymptotically large CO(2) (-)(H(2)O)(m) clusters is determined to be 0.466+/-0.001 eV (45.0+/-0.1 kJ/mol). This result compares very favorably with the heat of evaporation of bulk water, 0.456 eV (43.98 kJ/mol). PMID- 17461621 TI - Yield of excited CO molecules from dissociative recombination of HCO+ and HOC+ ions with electrons. AB - The authors have investigated CO band emissions arising from the dissociative recombination of HCO(+) and HOC(+) ions with thermal electrons in a flowing afterglow plasma. The quantitative analysis of the band intensities showed that HCO(+) recombination forms the long-lived CO(a (3)Pi) state with a yield of 0.23+/-0.12, while HOC(+) recombination favors formation of CO(a' (3)Sigma(+)) and CO(d (3)Delta) with a combined yield of greater than 0.4. The observed vibrational distribution for the CO(a) state reproduces theoretical predictions quite well. The vibrational distributions for CO(a') and CO(d) are, in part, inverted, presumably as a consequence of a change in CO equilibrium bond length during recombination. The observations are compatible with current knowledge of the potential surfaces of states of HCO and HCO(+). PMID- 17461622 TI - Yield of electronically excited CN molecules from the dissociative recombination of HNC+ with electrons. AB - The authors have studied CN(B-X) and CN(A-X) emissions produced by the dissociative recombination of HNC+ ions with thermal electrons in a flowing afterglow experiment. A separate drift tube study showed that the reaction Ar(+)+HCN, the precursor reaction used in the flow-tube experiment, produces predominantly HNC+ rather than the more energetic HCN+ isomer. Models simulating the ion-chemical processes, diffusion, and gas mixing in the afterglow plasma were fitted to observed position dependent CN(A-X) and CN(B-X) band intensities. Absolute yields of CN(B) and CN(A) were then obtained by comparing the CN band intensities to those of CO bands produced by recombination of CO(2) (+) ions. It was concluded that the 300 K recombination coefficient of HNC+ is close to 2 x 10(-7) cm(3) s(-1), that CN(B) is formed with a yield of 0.22+/-0.08 and CN(A) with a yield of 0.14+/-0.05. By comparison to synthetic spectra, the rotational temperature of CN(B) was estimated to be approximately 2500 K. It was also found that recombination produces CN(B) and CN(A) with far greater vibrational excitation than would be expected from the "impulse model" of Bates [Mon. Not. R. Astron. Soc. 263, 369 (1993)]. PMID- 17461623 TI - Jahn-Teller effect in CH3D+ and CD3H+: conformational isomerism, tunneling rotation structure, and the location of conical intersections. AB - High-resolution pulsed-field-ionization zero-kinetic-energy photoelectron spectra of CH(3)D and CD(3)H have been recorded at rotational resolution from the adiabatic ionization energy up to 600 cm(-1) of internal energy of the respective cations. The spectra are characterized by the effects of a large-amplitude pseudorotational motion exchanging the equivalent nuclei in each molecule. With increasing internal energy, a transition from the tunneling regime with splittings of the order of 1-10 cm(-1) to the free pseudorotation regime is observed. A theoretical model that treats the simultaneous rotational and pseudorotational motions and incorporates the effects of the geometric phase has been developed. The model provides the appropriate rovibronic symmetries in the C(3v)(M) molecular symmetry group and reaches a near-quantitative agreement with the experimental data. The complete group-theoretical analysis of the rovibronic problem is also given. The analysis of the spectra has revealed the existence of two different isomers for both CH(3)D(+) and CD(3)H(+), which differ in the bond length between the carbon atom and the unique ligand atom. All isomers are subject to a fast pseudorotational motion between three equivalent minima with a period of 3-5 ps in CH(3)D(+) and 18-28 ps in CD(3)H(+). The analysis has also provided the ordering of the tunneling sublevels for each isomer, which enables the location of the twofold conical intersections on the potential energy surface that could not be determined from experiments on CH(4) (+). PMID- 17461624 TI - Ab initio study of the torsional potential energy surfaces of N2O3 and N2O4: origin of the torsional barriers. AB - Intrinsic reaction coordinate (IRC) torsional potentials were calculated for N(2)O(4) and N(2)O(3) based on optimized B3LYP/aug-cc-pVDZ geometries of the respective 90 degrees -twisted saddle points. These potentials were refined by obtaining CCSD(T)aug-cc-pVXZ energies [in the complete basis set (CBS) limit] of points along the IRC. A comparison is made between these ab initio potentials and an analytical form based on a two-term cosine expansion in terms of the N-N dihedral angle. The shapes of these two potential curves are in close agreement. The torsional barriers in N(2)O(4) and N(2)O(3) obtained from the CCSD(T)/CBS//B3LYP/aug-cc-pVDZ calculations are 2333 and 1704 cm(-1), respectively. For N(2)O(4) the torsion fundamental frequency from the IRC potential is 87.06 cm(-1), which is in good agreement with the experimentally reported value of 81.73 cm(-1). However, in the case of N(2)O(3) the torsional frequency found from the IRC potential, 144 cm(-1), is considerably larger than the reported experimental values 63-76 cm(-1). Consistent with this discrepancy, the torsional barrier obtained from several different calculations, 1417-1718 cm( 1), is higher than the value of 350 cm(-1) deduced from experimental studies. It is suggested that the assignment of the torsional mode in N(2)O(3) should be reexamined. N(2)O(4) and N(2)O(3) exhibit strong hyperconjugative interactions of in-plane O lone pairs with the central N-N sigma* antibond. Hyperconjugative stabilization is somewhat stronger at the planar geometries because 1,4 interactions of lone pairs on cis O atoms promote delocalization of electrons into the N-N antibond. Calculations therefore suggest that the torsional barriers in these molecules arise principally from a combination of 1,4 interactions and hyperconjugation. PMID- 17461625 TI - Close-coupling study of rotational energy transfer and differential scattering in H2O collisions with He atoms. AB - Quantum close-coupling scattering calculations of rotational energy transfer (RET) of rotationally excited H(2)O due to collisions with He are presented for collision energies between 10(-6) and 1000 cm(-1) with para-H(2)O initially in levels 1(1,1), 2(0,2), 2(1,1), and 2(2,0) and ortho-H(2)O in levels 1(1,0), 2(1,2), and 2(2,1). Quenching cross sections and rate coefficients for state-to state RET were computed. Both elastic and inelastic differential cross sections are also calculated and compared with relative experimental results giving generally good agreement in all cases, but less so for inelastic results. Significant differences in the computed collisional parameters, obtained on three different potential energy surfaces (PESs), were found particularly in the ultracold regime. In the thermal regime, the rate coefficients calculated on each of the surfaces are generally in better agreement and comparable, but typically larger, than those obtained in a previous calculation. Unfortunately, a lack of absolute differential or integral inelastic experimental data prevents firm determination of a preferred PES. PMID- 17461626 TI - Bright, guided molecular beam with hydrodynamic enhancement. AB - The authors realize a novel high flux source of cold atoms and molecules employing hydrodynamic enhancement of an effusive aperture at cryogenic temperatures. Molecular oxygen from the source is coupled to a magnetic guide, delivering a cold, continuous, guided flux of 3 x 10(12) O(2) s(-1). The dynamics of the source are studied by creating and spectroscopically analyzing high flux beams of atomic ytterbium. PMID- 17461627 TI - Asymmetrically solvated anion with both kinetic and thermodynamic stabilities: Theoretical studies on the cluster anions (HF)n - (n=3-6). AB - At the level of MP2 with the aug-cc-pVDZ and aug-cc-pVTZ basis sets supplemented with diffuse bond functions, the authors searched the potential energy surfaces of (HF)(n) (-) (n=3-6). In accordance with the literature, they found that the symmetrically solvated-electron anion (3(FH){e}) possesses the largest vertical detachment energy (VDE), while the dipole-bound anion ((FH)(3){e}) is the lowest isomer in energy for (HF)(3) (-). Their calculations demonstrated that, with the increase of the cluster size, the asymmetric (FH)(a){e}(HF)(b) cluster is stabilized with a simultaneously increased VDE. Thus they predicted that, for (HF)(6) (-), the (FH)(4){e}(HF)(2) cluster is both kinetically and thermodynamically most stable, possessing the largest VDE and being the global minimum at the same time. PMID- 17461628 TI - Renner-Teller/Jahn-Teller intersections along the collinear axes of polyatomic molecules: C2H2 + as a case study. AB - Recently we discussed the Renner-Teller effect in triatomic molecules [J. Chem. Phys. 125, 094102 (2006)]. In that article the main message is that the Renner Teller phenomenon, just like the Jahn-Teller phenomenon, is a topological effect. Now we extend this study to a tetra-atomic system, namely, the C(2)H(2) (+) ion, for which topological effects are revealed when one atom surrounds the triatom axis or when two atoms surround (at a time) the two-atom axis. The present study not only supports the findings of the previous study, in particular, the crucial role played by the topological D matrix for diabatization, but it also reveals new features which are expected to be more and more pronounced the larger the original collinear molecule. As already implied, shifting away two atoms from the collinear molecular axis does not necessarily abolish the ability of the remaining two atoms to form topological effects. Moreover, the study indicates that when the two hydrogens are shifted away, the CC axis produces two kinds of topological effects: (1) a Renner-Teller effect (characterized by a topological phase of 2pi) which is revealed when the two hydrogens surround, rigidly, this axis (as mentioned above), and (2) a Jahn-Teller effect (characterized by a topological phase of pi) which is revealed when one of the hydrogens surrounds this axis while the other hydrogen is clamped to its position. PMID- 17461629 TI - Measurement of the partial photoionization cross sections and asymmetry parameters of S atoms in the photon energy range 10.0-30.0 eV using constant ionic-state spectroscopy. AB - The partial photoionization cross sections and asymmetry parameters of S atoms have been measured using constant-ionic-state (CIS) spectroscopy in the photon energy range 10.0-30.0 eV. The ionizations investigated in these CIS experiments are the (3p)(-1) ionizations S(+)((4)S)<--S((3)P), S(+)((2)D)<--S((3)P), and S(+)((2)P)<--S((3)P). For the first time Rydberg series which converge to the fourth ionization limit have been observed and assignments of these series have been proposed. These correspond to excitations to Rydberg states that are parts of series which converge to the fourth ionization limit, S(+)((4)P)<--S((3)P) (3s)(-1), and autoionize to the lower S(+)((4)S), S(+)((2)D), or S(+)((2)P) states. For each series observed in the CIS spectra photoelectron angular distribution studies, combined with other evidence, has allowed the angular momentum character of the free electron on autoionization to be determined. PMID- 17461630 TI - Ab initio investigation of the NH(X)-N2 van der Waals complex. AB - The NH-N(2) van der Waals complex has been examined at the CCSD(T) level of theory using aug-cc-pVDZ and aug-cc-pVTZ basis sets. The full basis set superposition error correction was applied. Two minimum energy structures were located for the electronic ground state. The global minimum corresponds to a linear geometry of the complex (NH-N-N), with D(e)=236 cm(-1) and R(c.m.)=4.22 A. The secondary minimum corresponds to a T-shaped geometry of C(2v) symmetry, where the nitrogen atom of the H-N moiety points toward the center of mass of the N(2) unit, aligned with the a-inertial axis of the complex. The binding energy and R(c.m.) value for the secondary minimum were 144 cm(-1) and 3.63 A, respectively. This potential energy surface is consistent with the properties of matrix isolated NH-N(2), and it is predicted that linear NH-N(2) will be a stable complex in the gas phase at low temperatures. PMID- 17461631 TI - Laser spectroscopy and dynamics of the jet-cooled AsH2 free radical. AB - The A (2)A(1)-X (2)B(1) electronic transition of the jet-cooled AsH(2) free radical has been studied by laser-induced fluorescence (LIF), wavelength-resolved emission, and fluorescence lifetime measurements. The radical was produced by a pulsed electric discharge through a mixture of arsine (AsH(3)) and high pressure argon at the exit of a pulsed valve. Nine vibronic bands were identified by LIF spectroscopy in the 505-400 nm region, including a long progression in the bending mode and two bands (1(0) (1) and 1(0) (1)2(0) (1)) involving the excited state As-H symmetric stretch. Single vibronic level emission spectra showed similar activity in the bending and symmetric stretching frequencies of the ground state. High-resolution spectra of the 0(0) (0) band exhibited large spin splittings and small, resolved arsenic hyperfine splittings, due to a substantial Fermi contact interaction in the excited state. The rotational constants obtained in the analysis gave effective molecular structures of r"(0)=1.5183(1) A, theta"(0)=90.75(1) degrees and r'(0)=1.4830(1) A, theta'(0)=123.10(2) degrees . The excited state fluorescence lifetimes vary dramatically with rovibronic state, from a single value of 1.4 micros to many with lifetimes less than 10 ns, behavior which the authors interpret as signaling the onset of a predissociative process near the zero-point level of the ground state. PMID- 17461632 TI - Computational study on the negative electron affinities of NO2 -.(H2O)n clusters (n=0-30). AB - Here we report negative electron affinities of NO(2)(-).(H2O)n clusters (n=0-30) obtained from density functional theory calculations and a simple correction to Koopmans' theorem. The method relies on the calculation of the detachment energy of the monoanion and its highest occupied molecular orbital and lowest unoccupied molecular orbital energies, and explicit calculations on the dianion itself are avoided. A good agreement with resonances in the cross section for neutral production in electron scattering experiments is found for n=0, 1, and 2. We find several isomeric structures of NO(2)(-).(H2O)2 of similar energy that elucidate the interplay between water-water and ion-water interactions. The topology is predicted to influence the electron affinity by 0.5 and 0.4 eV for NO(2)(-).(H2O) and NO(2)(-).(H2O)2, respectively. The electron affinity of larger clusters is shown to follow a (n+delta)-1/3 dependence, where delta=3 represents the number of water molecules that in volume, could replace NO(2) (-). PMID- 17461633 TI - Photoelectron imaging of copper and silver mono- and diamine anions. AB - The results of photoelectron imaging experiments of Cu and Ag mono- and diamine anions are reported. The photoelectron images were recorded at two photon energies, 800 and 527 nm. The vertical detachment energies of CuNH(2) (-) and AgNH(2) (-) are lower than those of the respective atomic metal ion and are measured to be 1.11+/-0.05 and 1.23+/-0.05 eV, respectively. By contrast, the electron detachment energies for Cu(NH(2))(2) (-) and Ag(NH(2))(2) (-) are higher than those of the corresponding metal ion and are determined to be 1.48+/-0.05 and 1.85+/-0.05 eV, respectively. Energy-dependent photoelectron anisotropy parameters are also reported. The photodetachment of the Cu and Ag mono- and diamine anions exhibit a cos(2) theta angular dependence relative to the direction of the laser polarization. The nature of the chemical bonding and the symmetry of the highest occupied molecular orbitals are discussed in relevance to the measured anisotropy parameters. PMID- 17461634 TI - Hydrogen peroxide and ammonia on protonated ice clusters. AB - A temperature controlled source for protonated water clusters has been combined with high-resolution mass spectroscopy to study the stability pattern of ice clusters and compounds with ammonia and hydrogen peroxide depending on temperature. The stability pattern of pure protonated ice shows the two well known peaks at 21 and 28 molecules and also less pronounced structure up to n=55. Ammonia and hydrogen peroxide do not destroy this pattern but shift it by a number of water molecules. The additives are therefore integrated in the persisting crystalline structure of the pure protonated ice. Based on this structural information, density functional theory calculations reveal that hydrogen peroxide and ammonia occupy surface positions on a dodecahedral 21 molecule cluster and are not caged in the center. PMID- 17461635 TI - Hyperpolarizability of GaAs dimer is not negative. AB - We present a systematic study of the static electric hyperpolarizability of Ga(2)As(2). The authors rely on finite-field high-level ab initio calculations with carefully optimized basis sets. Their best values for the mean and the anisotropy of the dipole polarizability are alpha=158.57 and Deltaalpha=130.33e(2)a(0) (2)E(h) (-1). For the hyperpolarizability we propose an estimate gamma=(155+/-15)x10(3)e(4)a(0) (4)E(h) (-3), which does not agree with the negative value predicted by Lan et al. [J. Chem. Phys. 124, 094302 (2006)]. Density functional theory based methods yield values close to those predicted by conventional ab initio methods. The (hyper)polarizability components are particularly enhanced along the direction defined by the Ga-Ga axis. PMID- 17461636 TI - Photodissociation yield spectroscopy of vinyl bromide cation generated by mass analyzed threshold ionization: vibrational spectroscopy and decay dynamics in the (~)B state. AB - A new technique [mass-analyzed threshold ionization (MATI)-photodissociation yield spectroscopy] to probe bound excited states of a cation was developed, which measures photodissociation yield of the cation generated by mass-analyzed threshold ionization. A vibrational spectrum of vinyl bromide cation in the (~)B state was obtained using this technique. Optical resolution in the low vibrational energy range of the spectrum was far better than in conventional MATI spectra. The origin of the (~)B state was found at 2.2578+/-0.0003 eV above the first ionization onset. Almost complete vibrational assignment was possible for peaks appearing in the spectrum. Analysis of time-of-flight profiles of C(2)H(3) (+) product ion obtained with different laser polarization angles suggested that photoexcited vinyl bromide cation remained in the (~)B state for several hundred picoseconds prior to internal conversion to the ground state and dissociation therein. PMID- 17461637 TI - Theoretical study of the electronic structure of MCH(2)(+)(M=Fe,Co,Ni). AB - State of the art coupled cluster (CC) methods are applied to accurately characterize the ground state electronic structure and photoelectron spectra of transition metal carbene ions MCH(2) (+) (M=Fe, Co, and Ni). The geometries and energies of the lowest energy quartet, triplet, and doublet electronic states as well as several low-lying vertical excitation energies of FeCH(2) (+), CoCH(2) (+), and NiCH(2) (+) are reported. The excitation energies are computed using the equation-of-motion CC and for states of different symmetries, by the energy differences of single reference ground and excited states (Delta-CC). The latter employ several reference states; the unrestricted Hartree-Fock, restricted open shell Hartree-Fock, and unrestricted Kohn-Sham. We conclude that the (2)A(1) electronic ground state of NiCH(2) (+) is separated by about 30.0 kJ/mol from the next highest state, and the lowest (4)B(1) and (4)B(2) states of FeCH(2) (+) as well as the (3)A(2) and (3)A(1) states of CoCH(2) (+) are nearly degenerate. The presence of metal-pi*(MCH(2)) charge transfer states with significant oscillator strengths in the visible/near-UV energy domain of the theoretical spectra of FeCH(2) (+) and CoCH(2) (+) are at the origin of the photofragmentation of these compounds observed after irradiation between 310 and 360 nm. PMID- 17461638 TI - The weak hydrogen bond in the fluorobenzene-ammonia van der Waals complex: Insights into the effects of electron withdrawing substituents on pi versus in plane bonding. AB - The fluorobenzene-ammonia van der Waals complex has been studied using a combination of two-color resonance enhanced multiphoton ionization (REMPI) spectroscopy, counterpoise corrected RICC2 ab initio molecular orbital calculations, and multidimensional Franck-Condon analysis. The experimental REMPI spectrum is characterized by a dominant, blueshifted band origin, and weak activity in intermolecular vibrational modes. RICC2 geometry optimizations and numerical vibrational frequency calculations of the neutral ground and first excited states have been performed on a number of different structural isomers of the complex using basis sets ranging from augmented double-zeta to quadruple-zeta level. Ground state basis set superposition error corrected zero-point binding energies show the in-plane sigma complex, forming a pseudo-six-membered ring connecting the fluorine atom and ortho-hydrogen, to be consistently the most stable of all six conformations considered, at all levels of theory. Comparison of computed zero-point excitation energies for the most stable pi and sigma conformers with fluorobenzene show that the sigma complex is the only conformer predicted to exhibit a spectral blueshift upon electronic excitation. The computed neutral ground and first excited state geometries and frequencies were used to perform multidimensional Franck-Condon simulations of the S(1)-S(0) vibronic spectrum for each of the most stable conformers. These simulations yielded null spectra for transitions involving the most stable of the pi complexes, pi(bridge); a spectrum rich in strong intermolecular vibrational structure for the second of the pi complexes, in complete contrast to the experimental spectrum; and for the sigma complex, a spectrum exhibiting weak intermolecular activity in line with that observed experimentally. This last simulation allowed an almost complete vibrational assignment of the intermolecular structure in the REMPI spectrum. The agreement between computational results and experiment overwhelmingly favors assignment of the spectrum to the in-plane sigma complex. PMID- 17461639 TI - In search of CS2(H2O)(n=1-4) clusters. AB - Gaussian-3 and MP2/aug-cc-pVnZ methods have been used to calculate geometries and thermochemistry of CS(2)(H2O)n, where n=1-4. An extensive molecular dynamics search followed by optimization using these two methods located two dimers, six trimers, six tetramers, and two pentamers. The MP2/aug-cc-pVDZ structure matched best with the experimental result for the CS(2)(H2O) dimer, showing that diffuse functions are necessary to model the interactions found in this complex. For larger CS(2)(H2O)n clusters, the MP2/aug-cc-pVDZ minima are significantly different from the MP2(full)6-31G* structures, revealing that the G3 model chemistry is not suitable for investigation of sulfur containing van der Waals complexes. Based on the MP2/aug-cc-pVTZ free energies, the concentration of saturated water in the atmosphere and the average amount of CS(2) in the atmosphere, the concentrations of these clusters are predicted to be on the order of 10(5) CS(2)(H2O) clusters.cm(-3) and 10(2) CS(2)(H2O)(2) clusters.cm(-3) at 298.15 K. The MP2/aug-cc-pVDZ scaled harmonic and anharmonic frequencies of the most abundant dimer cluster at 298 K are presented, along with the MP2/aug-cc pVDZ scaled harmonic frequencies for the CS(2)(H(2)O)(n) structures predicted to be present in a low-temperature molecular beam experiment. PMID- 17461640 TI - Quantum study on the branching ratio of the reaction NO2+OH. AB - A reduced dimensionality (RD) approximation is developed for the title reaction which treats the angle of approach of the hydroxyl radical to the nitrogen dioxide molecule and the radial distance between the two species explicitly. All other degrees of freedom are treated adiabatically. Electronic structure calculations at the complete active space self-consistent field level are used to fit a potential energy surface (PES) in these two coordinates. Within this RD model the adiabatic capture centrifugal sudden approximation is used to calculate the high pressure limit rate constant. A correction for reflection from the PES due to rotationally nonadiabatic transitions is applied using the wave packet capture approximation. The branching ratio for the title reaction is calculated for the atmospherically significant temperature range of 200-400 K at 20 Torr without distinguishing between the conformers of HOONO. The result is k(HOONO)k(HNO(3) )=0.051 at 20 Torr and 300 K, which is in good agreement with the measured branching ratio between cis-cis-HOONO and nitric acid. This suggests that most of the different conformers of HOONO were converted to the most stable cis-cis conformer on the time scale of the measurements made. PMID- 17461641 TI - Rapid measurement of three-dimensional diffusion tensor. AB - In this article, the authors demonstrate a rapid NMR method to measure a full three-dimensional diffusion tensor. This method is based on a multiple modulation multiple echo sequence and utilizes static and pulsed magnetic field gradients to measure diffusion along multiple directions simultaneously. The pulse sequence was optimized using a well-known linear inversion metric (condition number) and successfully tested on both isotropic (water) and anisotropic (asparagus) diffusion systems. PMID- 17461642 TI - Ab initio molecular dynamics simulation of ionic liquids. AB - Ab initio Car-Parinnello molecular dynamics is used to simulate the structure and the dynamics of 1-butyl-3-methylimidazolium iodide ([bmim]I) ionic liquid at 300 K. Site-site pair correlation functions reveal that the anion has a strong interaction with any three C-H's of the imidazolium ring. The ring bends over and wraps around the anion such that the two nitrogen atoms take a distance to the anion. Electron donating butyl group contributes the electronic polarization in addition to geometrical (out-of-plane) polarization of the ring due to the liquid environment. This facilitates bending of the ring along the axis passing through nitrogen atoms. The average bending angle depends largely on the alkyl chain length and slightly on the anion type. Redistribution of electron density over the ring caused by the electron donating alkyl group provides additional independent evidence to the instability of lattice structure, hence the low melting point of the ionic liquid. Simulated viscosity and diffusion coefficients of [bmim]I are in quite agreement with the experiments. PMID- 17461643 TI - On the study of collective dynamics in supercooled liquids through the statistics of the isoconfigurational ensemble. AB - The use of the isoconfigurational ensemble to explore structure-dynamic correlations in supercooled liquids is examined. The statistical error of the dynamic propensity and its spatial distribution are determined. The authors present the spatial distribution of the particle non-Gaussian parameter as a measure of the intermittency with which particles exhibit their propensity for motion. The ensemble average of the direction of particle motion is introduced to establish the anisotropy of the dynamic propensity. PMID- 17461644 TI - Thermal conductivity of ordered molecular water. AB - The authors use molecular dynamics simulation to investigate the thermal transport characteristics of water with various degree of orientational and translational orders induced by the application of an electric field. The authors observe that the orientational ordering of the water dipole moments has a minor effect on the thermal conductivity. However, electric-field-induced crystallization and associated translational order result in approximately a three fold increase of thermal conductivity with respect to the base water, i.e., to values comparable with those characterizing ice crystal structures. PMID- 17461645 TI - Photonic superdiffusive motion in resonance line radiation trapping Partial frequency redistribution effects. AB - The relation between the jump length probability distribution function and the spectral line profile in resonance atomic radiation trapping is considered for partial frequency redistribution (PFR) between absorbed and reemitted radiation. The single line opacity distribution function [M. N. Berberan-Santos et al., J. Chem. Phys. 125, 174308 (2006)] is generalized for PFR and used to discuss several possible redistribution mechanisms (pure Doppler broadening; combined natural and Doppler broadening; and combined Doppler, natural, and collisional broadening). It is shown that there are two coexisting scales with a different behavior: the small scale is controlled by the intricate PFR details while the large scale is essentially given by the atom rest frame redistribution asymptotic. The pure Doppler and combined natural, Doppler, and collisional broadening are characterized by both small- and large-scale superdiffusive Levy flight behaviors while the combined natural and Doppler case has an anomalous small-scale behavior but a diffusive large-scale asymptotic. The common practice of assuming complete redistribution in core radiation and frequency coherence in the wings of the spectral distribution is incompatible with the breakdown of superdiffusion in combined natural and Doppler broadening conditions. PMID- 17461646 TI - Quantum state tomography for quadrupolar nuclei using global rotations of the spin system. AB - In this paper, we describe a quantum state tomography method based on global rotations of the spin system which, together with a coherence selection scheme, enables the complete density matrix reconstruction. The main advantage of this technique, in respect to previous proposals, is the use of much shorter rf pulses, which decreases significantly the time necessary for algorithm quantum state tomography. In this case, under adequate experimental conditions, the rf pulses correspond to simple spatial rotations of the spin states, and its analytical description is conveniently given in the irreducible tensor formalism. Simulated results show the feasibility of the method for a single spin 72 nucleus. As an experimental result, we exemplify the application of this method by tomographing the steps during the implementation of the Deutsch algorithm. The algorithm was implemented in a (23)Na quadrupole nucleus using the strongly modulated pulses technique. We also extended the tomography method for a 3 coupled homonuclear spin 12 system, where an additional evolution under the internal Hamiltonian is necessary for zero order coherences evaluation. PMID- 17461647 TI - Water as a molecular hinge in amidelike structures. AB - The authors have studied the reorientational dynamics of isolated water molecules in a solution of N,N-dimethylacetamide (DMA). From linear spectra, the authors find that the water in this solution forms double hydrogen bond connections to the DMA molecules, resulting in the formation of DMA-water-DMA complexes. The authors use polarization-resolved mid-infrared pump-probe spectroscopy on the water in these complexes to measure the depolarization of three distinct transition dipole moments, each with a different directionality relative to the molecular frame (OH stretch in HDO, symmetric and asymmetric stretch normal modes in H(2)O). By combining these measurements, the authors find that the system exhibits bimodal rotational dynamics with two distinct time scales: a slow (7+/-1 ps) reorientation of the entire DMA-water complex and a fast (0.5+/-0.2 ps) "hinging" motion of the water molecule around the axis parallel to the connecting hydrogen bonds. Additionally, the authors observe an exchange of energy between the two normal modes of H(2)O at a time scale of 0.8+/-0.1 ps and find that the vibrational excitation decays through the symmetric stretch normal mode with a time constant of 0.8+/-0.2 ps. PMID- 17461648 TI - Isotope quantum effects in the electron momentum density of water. AB - The isotope quantum effects in the ground-state electron momentum density of water are studied at temperatures ranging from 5 to 90 degrees C by combining Compton scattering experiments utilizing synchrotron radiation and computational analysis within density functional theory. We observe clear differences in the momentum density between normal and heavy water at room temperature, which are interpreted as predominantly reflecting intramolecular structural differences. The changes in the momentum density upon increasing the temperature are found to be larger for heavy than for normal water, which is attributed primarily to temperature-induced intramolecular structural effects. Both model computations and an ab initio approach qualitatively reproduce the changes in the momentum density as a function of temperature. PMID- 17461649 TI - Computer simulation of phase separation under a double temperature quench. AB - The authors numerically study a two-step quench process in an asymmetric binary mixture. The mixture is first quenched to an unstable state in the two-phase region. After a large phase-separated structure is formed, the authors again quench the system deeper. The second quench induces the formation of small secondary droplets inside the large domains created by the first quench. The authors characterize this secondary droplet growth in terms of the temperature of the first quench as well as the depth of the second one. PMID- 17461650 TI - First-principles molecular dynamics study on aqueous sulfuric acid solutions. AB - The properties of aqueous sulfuric acid have been studied employing density functional theory-based molecular dynamics simulations in conjunction with norm conserving pseudopotentials. The simulations were carried out for two different concentrations whose molar concentrations were fixed at 0.84 and 10.2 mol/l. The structural features of aqueous sulfuric acid solutions show a strong dependency on the concentration. The Grotthuss-type proton transfer mechanism is not effectively operative at the higher concentration because of the broken hydrogen bond network of water induced by ions generated by the dissociation of sulfuric acid. In addition, to evaluate electrical properties, we carried out a simulation that takes an electric field into account. Results are compared with those of the simulation undertaken with no external electric field. PMID- 17461651 TI - Infrared spectroscopy of acetone-hexane liquid mixtures. AB - Acetone and hexane mixtures covering the whole solubility range were studied by Fourier transform infrared attenuated total reflectance spectroscopy. Factor analysis separates the spectra into four principal factor spectra and multiplying factors. Those containing negative factors are abstract, but the spectra are real. A statistical distribution model of the molecules in the solutions rendered the factors real. From these we define the intermediate species that occur in a 1:2 molar ratio of acetone in hexane, present principally in the low acetone concentration regions, and in a 2:1 molar ratio of acetone in hexane, present principally in the higher acetone concentration region. However, except at the concentration range limits where only pure acetone and pure hexane are present, the four species are present over the whole solubility range. The IR spectra of the species indicated very little displacement of the CH stretch bands, HCH deformation bands, and CC stretch bands, although there are some small intensity variations. Most of the modifications are observed on the acetone C=O stretch band. From the gas phase position, a strong bathochromic shift of 19 cm(-1) of the pure liquid is assigned to dipole-dipole interactions. In the 2:1 groupings, the shift that decreases to 15 cm(-1) is due to the diminished dipole-dipole interactions. In the 1:2 groupings, no dipole-dipole interaction can exist, and the bathochromic displacement of 9 cm(-1) is attributed to van der Waals interactions. In the one acetone to two hexanes grouping, no dipole-dipole interaction can exist, and the bathochromic displacement of 9 cm(-1) is attributed to van der Waals interactions. From the statistical distribution of the molecules, we determine that mixtures of hexane and acetone form a random organization with no preferred association or complex. PMID- 17461652 TI - Low temperature phases of the Andelman-de Gennes model of chiral discrimination: rigorous results. AB - We determine the ordered low temperature phases of the Andelman-de Gennes model of chiral discrimination, using rigorous statistical mechanical methods. The system is considered in the close-packed regime, equivalent to placing the molecules at every site of a honeycomb lattice. If the system contains an equimolar mixture of each of a pair of enantiomers, we prove in general that a heterochiral phase (disfavoring enantiomeric segregation) as well as a homochiral phase (favoring the segregation) is possible, depending on the types of intermolecular interactions. We apply our general results to the specific examples of the interactions considered by Andelman and de Gennes and provide a comparison with their conjectures that were based on two-molecule partition functions. PMID- 17461653 TI - Modeling the early stages of self-assembly in nanophase materials. AB - The early stages of self-assembly of the elementary building blocks of nanophase materials are studied. The relative roles of entropic and energetic factors in determining the relative abundance of the final products present is analyzed using both a kinetic mean field model and a mesoscopic approach in which self assembly is viewed as an encounter-controlled process on a discrete lattice. The relevance of the results in zeolite synthesis in connection with the ordered liquid phases recently discovered in these materials is discussed. PMID- 17461654 TI - Orientation of pentacene molecules on SiO2: from a monolayer to the bulk. AB - Near edge x-ray absorption fine structure (NEXAFS) spectroscopy is used to study the orientation of pentacene molecules within thin films on SiO2 for thicknesses ranging from monolayers to the bulk (150 nm). The spectra exhibit a strong polarization dependence of the pi* orbitals for all films, which indicates that the pentacene molecules are highly oriented. At all film thicknesses the orientation varies with the rate at which pentacene molecules are deposited, with faster rates favoring a thin film phase with different tilt angles and slower rates leading to a more bulklike orientation. Our NEXAFS results extend previous structural observations to the monolayer regime and to lower deposition rates. The NEXAFS results match crystallographic data if a finite distribution of the molecular orientations is included. Damage to the molecules by hot electrons from soft x-ray irradiation eliminates the splitting between nonequivalent pi* orbitals, indicating a breakup of the pentacene molecule. PMID- 17461655 TI - Density functional theory analysis of the structural and electronic properties of TiO2 rutile and anatase polytypes: performances of different exchange-correlation functionals. AB - The two polymorphs of TiO2, rutile and anatase, have been investigated at the ab initio level using different Hamiltonians with all-electron Gaussian and projector augmented plane wave basis sets. Their equilibrium lattice parameters, relative stabilities, binding energies, and band structures have been evaluated. The calculations have been performed at the Hartree-Fock, density functional theory (DFT), and hybrid (B3LYP and PBE0) levels. As regards DFT, the local density and generalized gradient (PBE) approximations have been used. Our results show an excellent agreement with the experimental band structures and binding energies for the B3LYP and PBE0 functionals, while the best structural descriptions are obtained at the PBE0 level. However, no matter which Hamiltonian and method are used, anatase is found more stable than rutile, in contrast with recent experimental reports, although the relative stabilities of the two phases are very close to each other. Nevertheless, based on the overall results, the hybrid PBE0 functional appears as a good compromise to obtain an accurate description of both structural and electronic properties of solids. PMID- 17461656 TI - Dynamics of low-coordinated surface atoms on gold nanocrystallites. AB - The authors highlight the importance of transient configurations of atoms on the surface of nanocrystallites, and present methodologies for their investigation. A Monte Carlo method has been developed and is used to simulate the thermodynamic equilibrium of nanometer sized Au nanocrystallites, both free and supported on a MgO(100) surface. The authors find that appreciable numbers of atoms transiently occupy adatom positions on Au(111) facets, even at room temperature. This type of dynamically appearing site is usually neglected in relation to catalysis but may have a significant activity (for CO oxidation, for example). They also observe a complex solid-solid roughening transition which involves a variety of transient local atom configurations on the surface of nanocrystallites. PMID- 17461657 TI - Spectroscopic characterization of carbon chains in nanostructured tetrahedral carbon films synthesized by femtosecond pulsed laser deposition. AB - A comparative study of carbon bonding states and Raman spectra is reported for amorphous diamondlike carbon films deposited using 120 fs and 30 ns pulsed laser ablation of graphite. The presence of sp(1) chains in femtosecond carbon films is confirmed by the appearance of a broad excitation band at 2000-2200 cm(-1) in UV Raman spectra. Analysis of Raman spectra indicates that the concentrations of sp(1)-, sp(2)-, and sp(3)-bonded carbon are approximately 6%, approximately 43%, and approximately 51%, respectively, in carbon films prepared by femtosecond laser ablation. Using surface enhanced Raman spectroscopy, specific vibrational frequencies associated with polycumulene, polyyne, and trans-polyacetylene chains have been identified. The present study provides further insight into the composition and structure of tetrahedral carbon films containing both sp(2) clusters and sp(1) chains. PMID- 17461658 TI - Molecular dynamics study on ultrathin liquid water film sheared between platinum solid walls: liquid structure and energy and momentum transfer. AB - Molecular dynamics simulation has been performed on a liquid film that is sheared in between solid surfaces. As a shear is given to the liquid film, a Couette-like flow is generated in the liquid and energy conversion occurs from the macroscopic flow to the thermal energy, which is discharged back to the solid walls. In such a way, momentum and thermal energy fluxes are present simultaneously. And all these thermal and fluid phenomena take place in highly nonequilibrium state where thermal energy is not distributed equally to each degree of freedom of molecular motion in the vicinities of the solid-liquid interface. In the present paper, platinum and water are employed as solid and liquid, respectively. First, the structure and orientation of water molecules in the vicinities of the solid surfaces are analyzed and how these structure and orientation are influenced by the shear is considered. Based on this result, momentum and thermal energy transfer in the vicinities of and at the solid-liquid interfaces are investigated in detail. Results are compared with those of our previous study, in which monatomic and diatomic molecules are employed as liquid. PMID- 17461659 TI - Surface tension of the most popular models of water by using the test-area simulation method. AB - We consider the calculation of the surface tension from simulations of several models of water, such as the traditional TIP3P, SPC, SPC/E, and TIP4P models, and the new generation of TIP4P-like models including the TIP4P/Ew, TIP4P/Ice, and TIP4P/2005. We employ a thermodynamic route proposed by Gloor et al. [J. Chem. Phys. 123, 134703 (2005)] to determine the surface tension that involves the estimate of the change in free energy associated with a small change in the interfacial area at constant volume. The values of the surface tension computed from this test-area method are found to be fully consistent with those obtained from the standard mechanical route, which is based on the evaluation of the components of the pressure tensor. We find that most models do not reproduce quantitatively the experimental values of the surface tension of water. The best description of the surface tension is given by those models that provide a better description of the vapor-liquid coexistence curve. The values of the surface tension for the SPC/E and TIP4P/Ew models are found to be in reasonably good agreement with the experimental values. From the present investigation, we conclude that the TIP4P/2005 model is able to accurately describe the surface tension of water over the whole range of temperatures from the triple point to the critical temperature. We also conclude that the test area is an appropriate methodological choice for the calculation of the surface tension not only for simple fluids, but also for complex molecular polar fluids, as is the case of water. PMID- 17461660 TI - First-principles study of structural, electronic, and multiferroic properties in BiCoO3. AB - Electronic and magnetic properties of BiCoO(3) have been investigated using the ab initio density-functional calculations with local spin density approximation (LSDA) and LSDA+U methods. The structural stability and the origin of the multiferroism for ferroelectronic and ferromagnetic existence were addressed. It was shown that the stability of the C-type antiferromagnetic (C-AFM) structure is better than that of other possible configurations. The hybridization between Bi-O and Co-O with interplay and a local magnetic moment on the Co(3+) play important roles for the nature of the ferroelectricity and ferromagnetism. Theoretical calculations predict the insulating ground state with a band gap of 2.11 eV in the C-AFM ordering for BiCoO(3) originated from the antiferromagnetic interaction in the ab plane, which is in well agreement with experiments. PMID- 17461661 TI - Monte Carlo calculation of second and third virial coefficients of small-scale comb polymers on lattice. AB - This paper reports the first computational estimation of the comb polymers' third virial coefficients. The number of the chains in the comb polymers range from 5 to 11. An algorithm that counts the contributing terms of the third virial coefficients in an accelerated manner is presented along with its efficiency dependence on the polymers' size. In addition, the second virial coefficients are estimated for the comb polymers and compared to previously reported results. PMID- 17461662 TI - Screening effects on structure and diffusion in confined charged colloids. AB - Using molecular dynamics computer simulations we investigate structural and dynamic (diffusion) properties of charged colloidal suspension confined to narrow slit pores with structureless, uncharged walls. The system is modeled on an effective level involving only the macroions, which interact via a combination of a soft-sphere and a screened Coulomb potential. The aim of our study is to identify the role of the range of the macroion-macroion interaction controlled by the inverse Debye screening length, kappa. We also compare to bulk properties at the same chemical potential as determined in parallel grand canonical Monte Carlo simulations. Our results reveal a significant influence of the interaction range which competes, however, with the influence of density. At liquidlike densities a decrease of range yields a decreasing mobility (and a corresponding enhancement of local structure) in the bulk system, whereas the reverse effect occurs in narrow slits with thickness of a few particle diameter. These differences can be traced back to the confinement-induced, and kappa-dependent, reduction of overall density compared to the bulk reservoir. We also show that an increase of kappa softens the oscillations in the normal pressure as function of the wall separation, which is consistent with experimental observations concerning the influence of addition of salt. PMID- 17461663 TI - Hybrid method coupling fluctuating hydrodynamics and molecular dynamics for the simulation of macromolecules. AB - We present a hybrid computational method for simulating the dynamics of macromolecules in solution which couples a mesoscale solver for the fluctuating hydrodynamics (FH) equations with molecular dynamics to describe the macromolecule. The two models interact through a dissipative Stokesian term first introduced by Ahlrichs and Dunweg [J. Chem. Phys. 111, 8225 (1999)]. We show that our method correctly captures the static and dynamical properties of polymer chains as predicted by the Zimm model. In particular, we show that the static conformations are best described when the ratio sigma/b=0.6, where sigma is the Lennard-Jones length parameter and b is the monomer bond length. We also find that the decay of the Rouse modes' autocorrelation function is better described with an analytical correction suggested by Ahlrichs and Dunweg. Our FH solver permits us to treat the fluid equation of state and transport parameters as direct simulation parameters. The expected independence of the chain dynamics on various choices of fluid equation of state and bulk viscosity is recovered, while excellent agreement is found for the temperature and shear viscosity dependence of center of mass diffusion between simulation results and predictions of the Zimm model. We find that Zimm model approximations start to fail when the Schmidt number Sc < or approximately 30. Finally, we investigate the importance of fluid fluctuations and show that using the preaveraged approximation for the hydrodynamic tensor leads to around 3% error in the diffusion coefficient for a polymer chain when the fluid discretization size is greater than 50 A. PMID- 17461664 TI - Structure of flexible and semiflexible polyelectrolyte chains in confined spaces of slit micro/nanochannels. AB - Understanding the behavior of a polyelectrolyte in confined spaces has direct relevance in design and manipulation of microfluidic devices, as well as transport in living organisms. In this paper, a coarse-grained model of anionic semiflexible polyelectrolyte is applied, and its structure and dynamics are fully examined with Brownian dynamics (BD) simulations both in bulk solution and under confinement between two negatively charged parallel plates. The modeling is based on the nonlinear bead-spring discretization of a continuous chain with additional long-range electrostatic, Lennard-Jones, and hydrodynamic interactions between pairs of beads. The authors also consider the steric and electrostatic interactions between the bead and the confining wall. Relevant model parameters are determined from experimental rheology data on the anionic polysaccharide xanthan reported previously. For comparison, both flexible and semiflexible models are developed accompanying zero and finite intrinsic persistence lengths, respectively. The conformational changes of the polyelectrolyte chain induced by confinements and their dependence on the screening effect of the electrolyte solution are faithfully characterized with BD simulations. Depending on the intrinsic rigidity and the medium ionic strength, the polyelectrolyte can be classified as flexible, semiflexible, or rigid. Confined flexible and semiflexible chains exhibit a nonmonotonic variation in size, as measured by the radius of gyration and end-to-end distance, with changing slit width. For the semiflexible chain, this is coupled to the variations in long-range bond vector correlation. The rigid chain, realized at low ionic strength, does not have minima in size but exhibits a sigmoidal transition. The size of confined semiflexible and rigid polyelectrolytes can be well described by the wormlike chain model once the electrostatic effects are taken into account by the persistence length measured at long length scale. PMID- 17461665 TI - Automatic discovery of metastable states for the construction of Markov models of macromolecular conformational dynamics. AB - To meet the challenge of modeling the conformational dynamics of biological macromolecules over long time scales, much recent effort has been devoted to constructing stochastic kinetic models, often in the form of discrete-state Markov models, from short molecular dynamics simulations. To construct useful models that faithfully represent dynamics at the time scales of interest, it is necessary to decompose configuration space into a set of kinetically metastable states. Previous attempts to define these states have relied upon either prior knowledge of the slow degrees of freedom or on the application of conformational clustering techniques which assume that conformationally distinct clusters are also kinetically distinct. Here, we present a first version of an automatic algorithm for the discovery of kinetically metastable states that is generally applicable to solvated macromolecules. Given molecular dynamics trajectories initiated from a well-defined starting distribution, the algorithm discovers long lived, kinetically metastable states through successive iterations of partitioning and aggregating conformation space into kinetically related regions. The authors apply this method to three peptides in explicit solvent-terminally blocked alanine, the 21-residue helical F(s) peptide, and the engineered 12 residue beta-hairpin trpzip2-to assess its ability to generate physically meaningful states and faithful kinetic models. PMID- 17461666 TI - Hierarchical analysis of conformational dynamics in biomolecules: transition networks of metastable states. AB - Molecular dynamics simulation generates large quantities of data that must be interpreted using physically meaningful analysis. A common approach is to describe the system dynamics in terms of transitions between coarse partitions of conformational space. In contrast to previous work that partitions the space according to geometric proximity, the authors examine here clustering based on kinetics, merging configurational microstates together so as to identify long lived, i.e., dynamically metastable, states. As test systems microsecond molecular dynamics simulations of the polyalanines Ala(8) and Ala(12) are analyzed. Both systems clearly exhibit metastability, with some kinetically distinct metastable states being geometrically very similar. Using the backbone torsion rotamer pattern to define the microstates, a definition is obtained of metastable states whose lifetimes considerably exceed the memory associated with interstate dynamics, thus allowing the kinetics to be described by a Markov model. This model is shown to be valid by comparison of its predictions with the kinetics obtained directly from the molecular dynamics simulations. In contrast, clustering based on the hydrogen-bonding pattern fails to identify long-lived metastable states or a reliable Markov model. Finally, an approach is proposed to generate a hierarchical model of networks, each having a different number of metastable states. The model hierarchy yields a qualitative understanding of the multiple time and length scales in the dynamics of biomolecules. PMID- 17461667 TI - Variable-free exploration of stochastic models: a gene regulatory network example. AB - Finding coarse-grained, low-dimensional descriptions is an important task in the analysis of complex, stochastic models of gene regulatory networks. This task involves (a) identifying observables that best describe the state of these complex systems and (b) characterizing the dynamics of the observables. In a previous paper [R. Erban et al., J. Chem. Phys. 124, 084106 (2006)] the authors assumed that good observables were known a priori, and presented an equation-free approach to approximate coarse-grained quantities (i.e., effective drift and diffusion coefficients) that characterize the long-time behavior of the observables. Here we use diffusion maps [R. Coifman et al., Proc. Natl. Acad. Sci. U.S.A. 102, 7426 (2005)] to extract appropriate observables ("reduction coordinates") in an automated fashion; these involve the leading eigenvectors of a weighted Laplacian on a graph constructed from network simulation data. We present lifting and restriction procedures for translating between physical variables and these data-based observables. These procedures allow us to perform equation-free, coarse-grained computations characterizing the long-term dynamics through the design and processing of short bursts of stochastic simulation initialized at appropriate values of the data-based observables. PMID- 17461669 TI - The effect of viewpoint on perceived visual roughness. AB - In previous work, we examined how the apparent roughness of a textured surface changed with direction of illumination. We found that observers exhibited systematic failures of roughness constancy across illumination conditions for triangular-faceted surfaces where physical roughness was defined as the variance of facet heights. These failures could be due, in part, to cues in the scene that confound changes in surface roughness with changes in illumination. These cues include the following: (1) the proportion of the surface in shadow, (2) mean luminance of the nonshadowed portion, (3) the standard deviation of the luminance of the nonshadowed portion, and (4) texture contrast. If the visual system relied on such "pseudocues" to roughness, then it would systematically misestimate surface roughness with changes in illumination much as our observers did despite the availability of depth cues such as binocular disparity. Here, we investigate observers' judgments of roughness when illumination direction and surface orientation are fixed and the observers' viewpoint with respect to the surface changes. We find a similar pattern of results. Observers exhibited patterned failures of roughness constancy with change in viewpoint, and an appreciable part of their failures could be accounted for by the same pseudocues. While the human visual system exhibits some degree of roughness constancy, our results lead to the conclusion that it does not always select the correct cues for a given visual task. PMID- 17461670 TI - Tuning for temporal interval in human apparent motion detection. AB - Detection of apparent motion in random dot patterns requires correlation across time and space. It has been difficult to study the temporal requirements for the correlation step because motion detection also depends on temporal filtering preceding correlation and on integration at the next levels. To specifically study tuning for temporal interval in the correlation step, we performed an experiment in which prefiltering and postintegration were held constant and in which we used a motion stimulus containing coherent motion for a single interval value only. The stimulus consisted of a sparse random dot pattern in which each dot was presented in two frames only, separated by a specified interval. On each frame, half of the dots were refreshed and the other half was a displaced reincarnation of the pattern generated one or several frames earlier. Motion energy statistics in such a stimulus do not vary from frame to frame, and the directional bias in spatiotemporal correlations is similar for different interval settings. We measured coherence thresholds for left-right direction discrimination by varying motion coherence levels in a Quest staircase procedure, as a function of both step size and interval. Results show that highest sensitivity was found for an interval of 17-42 ms, irrespective of viewing distance. The falloff at longer intervals was much sharper than previously described. Tuning for temporal interval was largely, but not completely, independent of step size. The optimal temporal interval slightly decreased with increasing step size. Similarly, the optimal step size decreased with increasing temporal interval. PMID- 17461671 TI - Some observations on the pedestal effect. AB - The pedestal or dipper effect is the large improvement in the detectability of a sinusoidal grating observed when it is added to a masking or pedestal grating of the same spatial frequency, orientation, and phase. We measured the pedestal effect in both broadband and notched noise-noise from which a 1.5-octave band centered on the signal frequency had been removed. Although the pedestal effect persists in broadband noise, it almost disappears in the notched noise. Furthermore, the pedestal effect is substantial when either high- or low-pass masking noise is used. We conclude that the pedestal effect in the absence of notched noise results principally from the use of information derived from channels with peak sensitivities at spatial frequencies different from that of the signal and the pedestal. We speculate that the spatial-frequency components of the notched noise above and below the spatial frequency of the signal and the pedestal prevent "off-frequency looking," that is, prevent the use of information about changes in contrast carried in channels tuned to spatial frequencies that are very much different from that of the signal and the pedestal. Thus, the pedestal or dipper effect measured without notched noise appears not to be a characteristic of individual spatial-frequency-tuned channels. PMID- 17461672 TI - Parameter learning but not structure learning: a Bayesian network model of constraints on early perceptual learning. AB - Visual scientists have shown that people are capable of perceptual learning in a large variety of circumstances. Are there constraints on such learning? We propose a new constraint on early perceptual learning, namely, that people are capable of parameter learning-they can modify their knowledge of the prior probabilities of scene variables or of the statistical relationships among scene and perceptual variables that are already considered to be potentially dependent but they are not capable of structure learning-they cannot learn new relationships among variables that are not considered to be potentially dependent, even when placed in novel environments in which these variables are strongly related. These ideas are formalized using the notation of Bayesian networks. We report the results of five experiments that evaluate whether subjects can demonstrate cue acquisition, which means that they can learn that a sensory signal is a cue to a perceptual judgment. In Experiment 1, subjects were placed in a novel environment that resembled natural environments in the sense that it contained systematic relationships among scene and perceptual variables that which are normally dependent. In this case, cue acquisition requires parameter learning and, as predicted, subjects succeeded in learning a new cue. In Experiments 2-5, subjects were placed in novel environments that did not resemble natural environments-they contained systematic relationships among scene and perceptual variables that are not normally dependent. Cue acquisition requires structure learning in these cases. Consistent with our hypothesis, subjects failed to learn new cues in Experiments 2-5. Overall, the results suggest that the mechanisms of early perceptual learning are biased such that people can only learn new contingencies between scene and sensory variables that are considered to be potentially dependent. PMID- 17461673 TI - Sequence learning in two-dimensional smooth pursuit eye movements in humans. AB - Sequence learning is common to all motor systems and is an essential aspect of human behavior necessary for the acquisition of motor skill. Many previous studies have demonstrated the ability to observe, store, and repeat sequences in a variety of modalities resulting in reduced reaction time. Recently, it has been found that subjects can make predictive smooth eye movements to a sequence of discrete horizontal target motions (C. J. Collins & Barnes, 2005). The present study extends that paradigm into two dimensions of motion in order to investigate qualitative and quantitative differences in sequences of vertical (V) and horizontal (H) eye movements. The subjects performed sequences of four discrete velocity ramps repeated either four or eight times in succession. Baseline measurements were obtained to discrete individual smooth pursuit velocity ramps to H and V predictable (PRD) and randomized (RND) targets. We found that subjects could rapidly learn and anticipate individual components of a four-ramp sequence in two dimensions. The results showed clear asymmetries in the eye movements made to horizontal and vertical targets. We found that the latencies to H targets were shorter than latencies to V targets in both the PRD and RND conditions. We also found higher initial eye velocity (50 ms after target onset) to H targets than vertical targets during the PRD condition. Because these differences in H and V eye movements are present in both RND and PRD trials, this suggests that the observed differences are not due to retention of information but are inherent asymmetries within the system. PMID- 17461674 TI - Electrophysiological correlates of perceptual reversals for three different types of multistable images. AB - Electrophysiological recordings were made in 21 observers to investigate whether differences in signature components (P1, N1, selection negativity [SN]) would be revealed during perceptual reversals of three different multistable figures. Using a lattice of Necker cubes as a stimulus, J. Kornmeier and M. Bach (2004, 2005) reported differences in P1 amplitudes as well a broad reversal-related negativity occurring 200-400 ms poststimulus. The current study investigated whether these event-related potentials of Necker cube reversals represent general "perceptual switching" mechanisms and would, therefore, be common to other types of multistable figures. Three different types of multistable stimuli were utilized: a modified Rubin's face/vase, a modified Schroder's staircase, and a novel natural stimulus, Lemmo's cheetahs. Results revealed the broad reversal related negativity for the face/vase and the reversible staircase but not for the cheetahs. This component is comparable to the SN in polarity, latency, and scalp topography. An effect of early visual spatial attention on figure reversals was suggested by an analysis of the occipital P1 and N1 components. The P1, N1, or both were enhanced for trials in which the observer reported perceptual reversals compared with trials in which no reversals were reported for the face/vase and reversible staircase stimuli. These results support a model of multistable perception in which changes in early spatial attention (indicated by P1 and N1 enhancement) modulate perceptual reversals (indicated by the reversal negativity or SN). PMID- 17461675 TI - What is the strength of a mask in visual metacontrast masking? AB - After more than a century of research, the mechanisms underlying visual masking are still hotly debated. One key characteristic of masking is that variations in the stimulus onset asynchrony (SOA) between the target and the mask can lead to either monotonic reductions in the effect of the mask on the target (A-type masking) or an increase in masking for intermediate SOAs and then a decrease in masking for longer SOAs (B-type masking). Past experimental and theoretical work suggested that the type of the masking function depends on the strength of the mask relative to the target. Usually, mask strength is related to energy (stimulus intensity x duration). Here, we show that the overall spatial layout of the mask is a much stronger factor than classical energy to explain the type of masking function. PMID- 17461676 TI - Modulation depth threshold in the Compensation Comparison approach. AB - Recently, the "Compensation Comparison" method was introduced for measuring retinal straylight. In this article, basic aspects are described, in particular a generalization of the approach using the concept of "precompensation," and including flicker threshold as parameter in the psychophysical model. The model was experimentally verified in lab measurements with and without artificially increased straylight and was tested on the data from the multi-center GLARE study. The resulting flicker threshold estimates were analyzed to better understand their origin. An effect of flicker adaptation over distance was found. The new approach proved suitable to describe Compensation Comparison measurements including precompensation, and also for subjects with poor psychometric behavior. PMID- 17461677 TI - Temporal aspects of orientation pooling using visual noise stimuli. AB - Previous psychophysical studies have shown that oriented signals are pooled over space to support better perceptual performance. We have investigated whether oriented signals may also be pooled over time. Using an orientation-in-noise paradigm, the threshold to discriminate large orientation differences was measured as the minimum amount of signal required to discriminate an oriented stimulus from unoriented noise. Discrimination thresholds were better with dynamic stimuli, containing multiple independent samples over time, than with static stimuli presented for an equal duration. Thresholds for dynamic stimuli showed gradual improvement from very brief (12 ms) to remarkably long presentation times (>4 s). Spatial integration of orientation signals is very efficient and can be understood in terms of a model based on signal detection theory, with performance limited by early and late stages of intrinsic noise. The nature of temporal integration is different, however, and is more consistent with probability summation of the outputs from low-level orientation detectors operating at a very brief timescale but whose outputs can be combined over a very long duration to yield better perceptual performance. PMID- 17461678 TI - What do we perceive in a glance of a real-world scene? AB - What do we see when we glance at a natural scene and how does it change as the glance becomes longer? We asked naive subjects to report in a free-form format what they saw when looking at briefly presented real-life photographs. Our subjects received no specific information as to the content of each stimulus. Thus, our paradigm differs from previous studies where subjects were cued before a picture was presented and/or were probed with multiple-choice questions. In the first stage, 90 novel grayscale photographs were foveally shown to a group of 22 native-English-speaking subjects. The presentation time was chosen at random from a set of seven possible times (from 27 to 500 ms). A perceptual mask followed each photograph immediately. After each presentation, subjects reported what they had just seen as completely and truthfully as possible. In the second stage, another group of naive individuals was instructed to score each of the descriptions produced by the subjects in the first stage. Individual scores were assigned to more than a hundred different attributes. We show that within a single glance, much object- and scene-level information is perceived by human subjects. The richness of our perception, though, seems asymmetrical. Subjects tend to have a propensity toward perceiving natural scenes as being outdoor rather than indoor. The reporting of sensory- or feature-level information of a scene (such as shading and shape) consistently precedes the reporting of the semantic-level information. But once subjects recognize more semantic-level components of a scene, there is little evidence suggesting any bias toward either scene-level or object-level recognition. PMID- 17461679 TI - Contrast thresholds for component motion with full and poor attention. AB - We compare luminance-contrast-masking thresholds for fully and poorly attended stimuli, controlling attention with a demanding concurrent task. We use dynamic displays composed of discrete spatiotemporal wavelets, comparing three conditions ("single," "parallel," and "random"). In contrast to static displays, we do not find that attention modulates the "dipper" regime for masks of low luminance contrast. Nor does attention alter direction-selective masking by multiple wavelets moving in random directions, a condition designed to isolate effects on component motion. However, direction-selective masking by multiple wavelets moving in parallel is significantly reduced by attention. As the latter condition is expected to excite both component and pattern motion mechanisms, this implies that attention may alter the visual representation of pattern motion. In addition, attention exhibits its well-known effect of reducing lateral masking between nearby spatiotemporal wavelets. PMID- 17461680 TI - Living up to optimal expectations. AB - Natural visual scenes contain several independent sources of information (cues) about a single property such as slant. It is widely assumed that the visual system processes such cues separately and then combines them with an averaging operation that takes the reliabilities of the individual cues into account. Does that mean that people lose access to information about inconsistencies between the cues, or are all inconsistencies revealed in a distorted surface appearance? To find out, we let observers match the slant and appearance of a simulated test surface to those of an identical, simultaneously visible, simulated reference surface and analyzed the variability in the settings. We also let observers match surfaces under conditions that were manipulated in ways that were expected to favor certain cues (monocular or binocular) or to selectively disrupt certain comparisons between the surfaces (slant or structure). The patterns in the variability between the settings were consistent with predictions based on the use of all available information. We argue that information about discrepancies is only "lost" during cue combination if there is no benefit in retaining the information. PMID- 17461681 TI - A model of spatiotemporal signal processing by primate cones and horizontal cells. AB - A model of spatiotemporal signal processing by the cone-horizontal cell circuit in the primate outer retina is developed and validated using measurements on the H1 horizontal cell from the literature. The model extends an earlier temporal model that mainly addressed the regulation of sensitivity by the cones. Three elements are added to the earlier model to describe the full spatiotemporal processing by horizontal cells. First, the feedback gain from horizontal cells to cones is made adaptive, depending on field size. Second, the spatial filtering by the horizontal dendritic tree is modeled as a two-component spatial filter. Third, an adaptive temporal low-pass filter is added, also depending on field size. The resulting model adequately describes all available measurements on spatiotemporal processing in macaque H1 cells. The adaptive feedback gain is argued to contribute to negative afterimages and chromatic adaptation in human vision. PMID- 17461682 TI - Human short-wavelength-sensitive cone light adaptation. AB - The cone-driven visual system is able to regulate its sensitivity effectively from twilight to bright sunlight. On the basis of a novel combination of short wavelength-sensitive (S-) cone measurements of temporal sensitivity and temporal delay, we show that S-cone light adaptation is achieved not only by trading unwanted sensitivity for speed but also by an additional process that counterintuitively increases the overall sensitivity as the light level rises. Our results are consistent with comparable middle-wavelength-sensitive (M-) cone measurements made in protanopic observers and can be accounted for by the same two-parameter model developed to account for the M-cone data (A. Stockman, M. Langendorfer, H. E. Smithson, & L. T. Sharpe, 2006). Each stage of the model can be linked to molecular mechanisms occurring within the photoreceptor: the speeding up to increases in the rates of decay of active and messenger molecules, the unexpected sensitivity increases to increased rates of molecular resynthesis and changes in channel sensitivity, and the sensitivity decreases to bleaching. Together, these mechanisms act to maintain vision in an optimal operating range and to protect it from overload. PMID- 17461684 TI - Where to look next? Eye movements reduce local uncertainty. AB - How do we decide where to look next? During natural, active vision, we move our eyes to gather task-relevant information from the visual scene. Information theory provides an elegant framework for investigating how visual stimulus information combines with prior knowledge and task goals to plan an eye movement. We measured eye movements as observers performed a shape-learning and -matching task, for which the task-relevant information was tightly controlled. Using computational models, we probe the underlying strategies used by observers when planning their next eye movement. One strategy is to move the eyes to locations that maximize the total information gained about the shape, which is equivalent to reducing global uncertainty. Observers' behavior may appear highly similar to this strategy, but a rigorous analysis of sequential fixation placement reveals that observers may instead be using a local rule: fixate only the most informative locations, that is, reduce local uncertainty. PMID- 17461683 TI - Glass pattern responses in macaque V2 neurons. AB - Area V2 of macaque visual cortex is known to respond well to conventional oriented bar and grating stimuli, but some recent physiological data have shown that it may play an important role in coding more complicated patterns. Most of these data come from testing done with stimuli presented within the classical receptive field (CRF), whereas relatively little attention has been paid to the role played by the extra-classical surround. We have previously shown that neurons in primary visual cortex (V1) respond to translational Glass patterns in a manner that is predictable from their responses to grating stimuli. In this article, we first extend our experiments and modeling of Glass pattern responses in V1 to include V2. We explored the sensitivity of V2 cells to global form cues in Glass patterns confined to the CRF. Our results indicate that V2 neurons respond to the local signals in Glass patterns in a manner similar to V1 and that those responses are not influenced by global form present in the surround. It appears that the coding of the more complicated global structure in Glass patterns takes place further downstream in the visual system. PMID- 17461685 TI - Simulating human cones from mid-mesopic up to high-photopic luminances. AB - A computational model of human cones for intensities ranging from 1 td up to full bleaching levels is presented. The model conforms well with measurements made in primate horizontal cells, follows Weber's law at high intensities, and performs range compression consistent with what is known of cones in other vertebrates. The model consists entirely of processes with a clear physiological interpretation: pigment bleaching, saturation of cGMP hydrolysis, calcium feedback on cGMP synthesis, and a nonlinear membrane. The model is implemented according to a very fast computational scheme useful for simulations, and sample programs in Matlab and Fortran are provided as supplementary material. PMID- 17461686 TI - Effects of spatial and temporal context on color categories and color constancy. AB - Color constancy is the ability to assign a constant color to an object independent of changes in illumination. Color constancy is achieved by taking context information into account. Previous approaches that have used matching paradigms to quantify color constancy found degrees of constancy between 20% and 80%. Here, we studied color constancy in a color-naming task under different conditions of surround illumination and patch size. Observers categorized more than 400 patches for each illumination condition. This allows one to overcome inherent limitations in color naming and to study the changes in color categories under illumination changes. When small central test patches with a full context illumination were categorized, observers followed the illumination shift almost completely, showing a high degree of constancy (99%). Reducing the available context information or increasing the patch size decreased the degree of constancy to about 50%. Moderate degrees of constancy (66%) occurred even when the test patches were never viewed simultaneously but only in temporal alternation with the illumination. Boundaries between color categories were largely stable within and across observers under neutral illumination. Under changing illumination, there were small but systematic variations in the color category boundaries. Color category boundaries tended to rotate away from the illumination color. This variation was largest under full context conditions where highest degrees of color constancy were obtained. PMID- 17461687 TI - Motion signals bias localization judgments: a unified explanation for the flash lag, flash-drag, flash-jump, and Frohlich illusions. AB - In the flash-lag illusion, a moving object aligned with a flash is perceived to be offset in the direction of motion following the flash. In the "flash-drag" illusion, a flash is mislocalized in the direction of nearby motion. In the "flash-jump" illusion, a transient change in the appearance of a moving object (e.g., color) is mislocalized in the direction of subsequent motion. Finally, in the Frohlich illusion, the starting position of a suddenly appearing moving object is mislocalized in the direction of the subsequent motion. We demonstrate, in a series of experiments, a unified explanation for all these illusions: Perceptual localization is influenced by motion signals collected over approximately 80 ms after a query is triggered. These demonstrations rule out "latency difference" and asynchronous feature binding models, in which objects appear in their real positions but misaligned in time. Instead, the illusions explored here are best understood as biases in localization caused by motion signals. We suggest that motion biasing exists because it allows the visual system to account for neural processing delays by retrospectively "pushing" an object closer to its true physical location, and we propose directions for exploring the neural mechanisms underlying the dynamic updating of location by the activity of motion-sensitive neurons. PMID- 17461688 TI - Added noise affects the neural correlates of upright and inverted faces differently. AB - In five experiments, we examine the neural correlates of the interaction between upright faces, inverted faces, and visual noise. In Experiment 1, we examine a component termed the N170 for upright and inverted faces presented with and without noise. Results show a smaller amplitude for inverted faces than upright faces when presented in noise, whereas the reverse is true without noise. In Experiment 2, we show that the amplitude reversal is robust for full faces but not eyes alone across all noise levels. In Experiment 3, we vary contrast to see if this reversal is a result of degrading a face. We observe no reversal effects. Thus, across conditions, adding noise to full faces is a sufficient condition for the N170 reversal. In Experiment 4, we delay the onsets of the faces presented in noise. We replicate the smaller N170 for inverted faces at no delay but observe partial recovery of the N170 for inverted faces at longer delays in static noise. Experiment 5 demonstrates the interaction in low contrast at a behavioral level. We propose a model in which noise interacts with the processing properties of inverted faces more so than upright faces. PMID- 17461689 TI - Monocular symmetry in binocular vision. AB - Human vision is highly sensitive to bilateral symmetry in 2-D images. It is, however, not clear yet whether this visual sensitivity relates to symmetry of 3-D objects or whether it relates to symmetry of the 2-D image itself. We used a stereoscopically presented stimulus and a 3-D bisection task that enable us to dissociate object symmetry from image symmetry. The bisection stimulus consisted of three parallel lines, of which two lines were located in one depth plane and the third line in another. Bisection judgments were different for horizontal and vertical lines, which can be explained by taking into account the distinct viewpoints of the left and right eyes for either of the visible sides of the 3-D object. Image symmetry from a monocular vantage point predicts 3-D bisection better than object symmetry. We conclude that observers use either of the two monocular 2-D images separately but not a single cyclopean view-nicely dovetailing with what they do when they assess both 3-D visual direction and 3-D shape-to assess 3-D symmetry. PMID- 17461690 TI - Silhouetted face profiles: a new methodology for face perception research. AB - We present a new methodology for constructing face stimuli for psychological experiments based on silhouetted face profiles. Face silhouettes carry a number of theoretical and methodological advantages compared to more complex face stimuli and lend themselves to a simple yet powerful parameterization. In five behavioral studies, we show that face silhouettes are processed like regular face stimuli: They provide enough information for accurate gender judgments (Study 1), age estimations (Study 2), and reliable and cross-valid attractiveness ratings (Study 3). Furthermore, face silhouettes elicit an inversion effect (Study 4) and allow for remarkably accurate cross-identification with front-view photographs (Study 5). We then describe a shape-based parameterization that relies on a small set of landmark points and show that face silhouettes can be effectively represented in a 20-dimensional "silhouette face space" (Study 6). We show that in this physical space, distance from the center of the space corresponds to perceived distinctiveness (Study 7), confirming a key axiom in the formulation of the face space model. Finally, we discuss straightforward applications of the face silhouette methodology and address some limitations. PMID- 17461691 TI - Contextual modulation involves suppression and facilitation from the center and the surround. AB - In psychophysics, cross-orientation suppression (XOS) and cross-orientation facilitation (XOF) have been measured by investigating mask configuration on the detection threshold of a centrally placed patch of sine-wave grating. Much of the evidence for XOS and XOF comes from studies using low and high spatial frequencies, respectively, where the interactions are thought to arise from within (XOS) and outside (XOF) the footprint of the classical receptive field. We address the relation between these processes here by measuring the effects of various sizes of superimposed and annular cross-oriented masks on detection thresholds at two spatial scales (1 and 7 c/deg) and on contrast increment thresholds at 7 c/deg. A functional model of our results indicates the following (1) XOS and XOF both occur for superimposed and annular masks. (2) XOS declines with spatial frequency but XOF does not. (3) The spatial extent of the interactions does not scale with spatial frequency, meaning that surround-effects are seen primarily at high spatial frequencies. (4) There are two distinct processes involved in XOS: direct divisive suppression and modulation of self suppression. (5) Whether XOS or XOF wins out depends upon their relative weights and mask contrast. These results prompt enquiry into the effect of spatial frequency at the single-cell level and place new constraints on image-processing models of early visual processing. PMID- 17461692 TI - Residual cone vision without alpha-transducin. AB - Behavioral experiments in humans with a rare genetic mutation that compromises the function of alpha-transducin (Galpha the alpha-subunit of the G-protein in the primary cone phototransduction cascade) reveal a residual cone response only viable at high light levels and at low temporal frequencies. It has three characteristic properties. First, it limits temporal frequency sensitivity to the equivalent of a simple first order reaction with a time constant of approximately 140 ms. Second, it delays the visual response by an amount that is also consistent with such a reaction. Third, it causes temporal acuity to be linearly related to the logarithm of the amount of bleached pigment. We suggest that these properties are consistent with the residual function depending on a sluggishly generated cone photobleaching product, which we tentatively identify as a cone metarhodopsin. By activating the transduction cascade, this bleaching product mimics the effects of real light and is therefore one of the molecular origins of "background equivalence," the long-established observation that the aftereffects of photopigment bleaches and the effects of real background lights are equivalent. Alternative explanations for the residual cone response include the possibilities that there is a secondary phototransduction mechanism that bypasses alpha-transduction, or that the truncated alpha-transduction that results from the mutation retains some minimal functionality. PMID- 17461693 TI - Orientation discrimination in 5-year-olds and adults tested with luminance modulated and contrast-modulated gratings. AB - We compared thresholds for discriminating orientation by 5-year-olds and adults for first-order (luminance-modulated) and second-order (contrast-modulated) gratings. To achieve equal visibility, we set the contrast for each age and condition at a fixed multiple of the contrast threshold for discriminating horizontal from vertical gratings. The minimum tilt that could be discriminated from vertical was four to five times larger in 5-year-olds than in adults, even when the noise was removed from the first-order stimuli and amplitude modulation increased to 0.90. Thresholds at both ages were significantly worse (1.2-1.5 times worse) for second-order modulation than for equally visible first-order modulation, and 5-year-olds were equally immature for both types of pattern. Together, the findings suggest that orientation discrimination is slow to develop and worse for second-order than first-order patterns in both children and adults. PMID- 17461694 TI - Fear perception: can objective and subjective awareness measures be dissociated? AB - Whereas previous studies of fearful-face perception have probed visual awareness according to either objective or subjective criteria, in the present study, we probed the perception of briefly presented and masked fearful faces by assessing both types of perception within the same task. Both objective and subjective sensitivity measures were assessed within a common signal detection theory framework. To evaluate single-participant awareness, we employed a nonparametric receiver operating characteristic (ROC) analysis of the behavioral data, which involved collecting a large number of trials over multiple sessions. Our findings revealed that nearly all subjects could reliably detect 17-ms fearful-face targets, thus exhibiting above-chance objective perception at this target duration. Reliable subjective sensitivity was also observed for 33-ms fearful face targets and, for some subjects, even for 17-ms targets. The analysis of single-session data suggests that previous experiments may have lacked sufficient statistical power to establish above-chance performance. Taken together, our findings are consistent with a dissociation of fear perception according to objective and subjective criteria, which could be assessed for each individual participant. The determination of such a dissociation zone may help in understanding the conditions linked to aware and unaware fear perception. PMID- 17461695 TI - Controversy and conflict in the treatment of acute decompensated heart failure: nesiritide as evidence-based treatment. AB - Acute decompensated heart failure poses a complex clinical challenge for the health care community. Evolving concepts of the pathophysiology and lack of consensus on appropriate outcome measures for drug approval underlie some of the current controversies about nesiritide. We outline the major controversies from the viewpoint that nesiritide should continue to be used judiciously by following its package insert recommendations and the Heart Failure Society of America's 2006 Comprehensive Heart Failure Practice Guidelines. PMID- 17461696 TI - Controversy and conflict in the treatment of acute decompensated heart failure: limited role for nesiritide. AB - The use of nesiritide for acute decompensated heart failure (ADHF) has been clouded with controversy since its approval in 2001. Extensive marketing and many review articles have established this drug as a safe and superior product to current standards. However, its safety has been called into question by the results of a meta-analysis, and its superiority of important outcomes (length of stay, mortality, decreased readmission rate) has never been proved by a randomized trial against agents with similar vasodilator properties (e.g., nitroglycerin). A review of the available literature on nesiritide in the areas of mortality, renal effects, retrospective studies, use in off-label indications, length of stay, and mortality is presented and illustrates why its use should be limited or even eliminated. After review of this article, the reader should be able to answer the question--if nesiritide had never been approved for use in patients with ADHF, would we have missed it?--with a negative reply. PMID- 17461697 TI - Effect of omega-3 fatty acid supplementation on the arachidonic acid:eicosapentaenoic acid ratio. AB - STUDY OBJECTIVES: To determine the baseline arachidonic acid:eicosapentaenoic acid (AA:EPA) ratio in patients with coronary artery disease and healthy subjects, and whether supplementation of omega-3 fatty acids, administered as fish oil capsules, affects this ratio. DESIGN: Prospective, open-label trial. SETTING: University-affiliated cardiology clinic. SUBJECTS: Thirty patients with stable coronary artery disease (CAD) and 30 healthy subjects. INTERVENTION: All participants received omega-3 fatty acids 1.5 g/day for 4 weeks, followed by 3 g/day for an additional 4 weeks. MEASUREMENTS AND MAIN RESULTS: For each participant, a lipid profile was determined at baseline and after 4 weeks of treatment with each dose. Other laboratory results analyzed were serum AA:EPA ratios, high-sensitivity C-reactive protein (hs-CRP) levels, and blood glucose levels. Mean +/- SD baseline AA:EPA ratios were 39.6 +/- 19.0 in healthy subjects and 23.7 +/- 12.5 in patients with CAD. These ratios decreased significantly in both groups after treatment with 1.5 g/day of omega-3 fatty acids: 9.0 +/- 4.2 in healthy subjects and 10.3 +/- 8.8 in patients with CAD. After treatment with 3 g/day, the ratios were further reduced: 5.1 +/- 3.2 in healthy subjects and 4.9 +/- 2.6 in patients with CAD. Supplementation with omega-3 fatty acids did not significantly affect hs-CRP, high-density lipoprotein cholesterol, low-density lipoprotein cholesterol, or blood glucose levels. Triglyceride levels were not reduced in patients with CAD but were significantly decreased in healthy subjects (by 20% decrease with omega-3 fatty acids 1.5 g/day and by 32% decrease with 3 g/day). CONCLUSION: Treatment with omega-3 fatty acids significantly reduced AA:EPA ratios in both healthy subjects and in patients with stable CAD. The treatment had no effect on hs-CRP levels in either group, and it reduced triglyceride levels in healthy subjects but not in patients with CAD. PMID- 17461698 TI - Development and reliability testing of the clinical pharmacist recommendation taxonomy. AB - STUDY OBJECTIVE: To evaluate the reliability of a newly developed taxonomy--the Clinical Pharmacist Recommendation (CPR) taxonomy--to classify clinical pharmacy interventions. DESIGN: The CPR taxonomy was developed and refined in three phases. In each phase, reviewers independently reviewed recommendations made by a clinical pharmacist-physician team and categorized them into mutually exclusive categories: priority, problem, and response. Interrater reliability was assessed for all categories during each development phase. SETTING: Primary care clinics of a Veterans Affairs Medical Center. PATIENTS: Fifty-three patients enrolled in the Veterans Affairs Enhanced Pharmacy Outpatient Clinic (EPOC) trial. MEASUREMENTS AND MAIN RESULTS: Interrater reliability was assessed using the kappa statistic. A total of 423 recommendations were evaluated during the three testing phases. In the final testing phase, agreement was moderate for pharmacotherapy problem subcategories (kappa = 0.57), substantial for pharmacotherapy problem primary categories (kappa = 0.64), and almost perfect for response categories (kappa = 0.85). Taxonomy completion time/patient averaged 4.6 minutes (range 1-11 min). CONCLUSION: The CPR taxonomy provides a reliable method to systematically evaluate clinical pharmacy recommendations based on the therapeutic problem identified and specific action recommended to resolve the problem. PMID- 17461699 TI - Comparison of patient outcomes with bivalirudin versus unfractionated heparin in percutaneous coronary intervention. AB - OBJECTIVE: To compare clinical outcomes and glycoprotein IIb-IIIa inhibitor use in patients undergoing percutaneous coronary intervention (PCI) who received bivalirudin or unfractionated heparin (UFH) in a real-world setting. DESIGN: Retrospective cohort analysis. SETTING: University-affiliated medical center. PATIENTS: One thousand seventy-five adult patients who underwent PCI and received either bivalirudin (539 patients) or UFH (536 patients) from April 1, 2003-April 1, 2004. MEASUREMENT AND MAIN RESULTS: Patient data on demographics, comorbidities, laboratory values, and reports of radiologic examinations, cardiac catheterizations, and discharge summaries were obtained. Outcomes evaluated included rates of in-hospital mortality, myocardial infarction, revascularization, and length of stay (LOS), as well as Randomized Evaluation of PCI Linking Angiomax to Reduced Clinical Events (REPLACE-2) and Thrombosis in Myocardial Infarction (TIMI) bleeding categorization. Bivalirudin use was associated with a significant reduction in TIMI major (5.0% vs 9.7%, p=0.003), REPLACE-2 major (5.4% vs 12.9%, p<0.001), and TIMI minor (1.7% vs 6%, p<0.001) bleeding complications compared with UFH use. Significantly fewer patients in the bivalirudin group received glycoprotein IIb-IIIa inhibitors (27.3% vs 62.7%, p<0.001). Patients receiving bivalirudin had significantly fewer myocardial infarctions after catheterization (10.7% [40/375] vs 18.0% [51/284], p=0.007). No differences were noted in mortality and revascularization rates between groups. A shortened LOS was observed in the bivalirudin group. CONCLUSIONS: This real-world analysis that included high-risk patients provides further evidence that bivalirudin is an attractive alternative to UFH because of a decrease in bleeding events without compromising efficacy. PMID- 17461700 TI - Low-dose botulinum toxin type A for the treatment of refractory piriformis syndrome. AB - STUDY OBJECTIVES: To evaluate the efficacy of a single, low-dose injection of botulinum toxin type A in relieving pain in Korean patients with piriformis syndrome resistant to conventional therapy, and to assess the drug's influence on these patients' quality of life. DESIGN: Prospective, single-site, open-label trial. SETTING: Rehabilitation medicine clinic in Seoul, Korea. PATIENTS: Twenty nine patients with a confirmed diagnosis of chronic piriformis syndrome and 82 age- and sex-matched healthy subjects were enrolled from April 1, 2003-February 28, 2004. Intervention. In 20 of the patients, botulinum toxin type A 150 U was injected using computed tomographic guidance into the affected unilateral piriformis muscle. The other nine patients served as active controls and received an injection of dexamethasone 5 mg and 1% lidocaine. The healthy subjects did not receive any injection. MEASUREMENTS AND MAIN RESULTS: The patients' pain at baseline and at 4, 8, and 12 weeks after treatment was rated by using a numeric rating scale. Health-related quality of life was assessed by using the validated Korean version of the Medical Outcomes Study 36-Item Short Form Health Survey (SF 36) at baseline and at 4 weeks of treatment. Healthy subjects also completed the SF-36 at baseline. Pain intensity scores were significantly lower at 4, 8, and 12 weeks after treatment than at baseline (p<0.0001). Baseline scores from the SF-36 subscales, including those for physical functioning (p<0.0001), role physical (p<0.0001), bodily pain (p<0.0001), general health (p<0.0001), vitality (p<0.0001), and social functioning (p<0.002), were significantly lower in the patients than in the healthy subjects. Four weeks after treatment, physical functioning (p=0.003), role physical (p=0.021), bodily pain (p=0.016), general health (p=0.013), vitality (p=0.031) and social functioning (p=0.035) improved significantly from baseline in the patients. However, at 4 weeks, patients in the active control group were withdrawn from the study because their pain did not improve, and continuation without further medical care was considered unethical. CONCLUSION: A low dose of botulinum toxin type A relieved pain and improved quality of life in patients with refractory piriformis syndrome. PMID- 17461701 TI - Prevalence of illicit use and abuse of prescription stimulants, alcohol, and other drugs among college students: relationship with age at initiation of prescription stimulants. AB - STUDY OBJECTIVE: To examine associations between age at initiation of prescription stimulants and illicit use and abuse of prescription stimulants, alcohol, and other drugs among college students in the United States. DESIGN: Web based survey of college students. SETTING: A large (full-time undergraduate population > 20,000) university. INTERVENTION: A Web-based survey was sent to a random sample of 5389 undergraduate college students plus an additional 1530 undergraduate college students of various ethnic backgrounds over a 2-month period. MEASUREMENTS AND MAIN RESULTS: Alcohol abuse was assessed by including a modified version of the Cut Down, Annoyance, Guilt, Eye-opener (CAGE) instrument. Drug use-related problems were assessed with a slightly modified version of the Drug Abuse Screening Test, short form (DAST-10). The final sample consisted of 4580 undergraduate students (66% response rate). For the analyses, five subgroups were created based on age at initiation of prescription stimulant use: no prescription stimulant use, grades kindergarten (K)-4, grades 5-8, grades 9-12, and college. Undergraduate students to whom stimulants were prescribed in grades K-4 reported similar rates of alcohol and other drug use compared with that of the group that had no prescription stimulant use. For example, students who started prescription stimulants in grades K-4 were no more likely to report coingestion of alcohol and illicit prescription stimulants (odds ratio [OR] 1.4, 95% confidence interval [CI] 0.2-11.5, NS] than the group that had no prescription stimulant use. However, undergraduate students whose prescription stimulant use began in college had significantly higher rates of alcohol and other drug use. For example, students who started a prescription stimulant in college were almost 4 times as likely (OR 3.7, 95% CI 1.9-7.1, p<0.001) to report at least three positive indicators of drug abuse on the DAST-10 compared with the group that had no prescription stimulant use. CONCLUSIONS: In concordance with results of previous research, these results indicate that initiation of prescription stimulants during childhood is not associated with increased future use of alcohol and other drugs. PMID- 17461702 TI - The effect of acetaminophen on the international normalized ratio in patients stabilized on warfarin therapy. AB - STUDY OBJECTIVE: To determine whether an interaction exists between acetaminophen and warfarin that alters the international normalized ratio (INR). DESIGN: Prospective, randomized, double-blind, placebo-controlled trial. SETTING: Anticoagulation clinic at a Veterans Affairs Medical Center. PATIENTS: Thirty-six adult patients receiving warfarin with stable INRs, defined as two consecutive INRs at least 3 weeks apart that were within the therapeutic range. INTERVENTION: Patients were randomly assigned to receive acetaminophen 1 g twice/day along with matching placebo twice/day (12 patients), acetaminophen 1 g 4 times/day (12 patients), or matching placebo 4 times/day (12 patients) for 4 weeks. MEASUREMENTS AND MAIN RESULTS: The primary end point was the difference in mean INR between groups at weekly intervals. Secondary end points were the percentages of patients in each group with supratherapeutic (INR > or = 0.3 above the upper limit of their therapeutic range) or subtherapeutic (INR > or = 0.2 or 0.3 below the lower limit of their respective therapeutic range of 2.0-3.0 or 2.5-3.5) INRs, and the difference in mean serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels between groups at biweekly intervals. Slow enrollment and a preliminary observation that 15 patients experienced an elevated INR prompted early termination of the study. At week 2, the group receiving acetaminophen 2 g/day had a significantly higher mean INR versus the placebo group (p=0.01). At weeks 1, 2, and 3, the acetaminophen 4-g/day group had significantly higher mean INRs compared with those in the placebo group (p=0.04, p=0.01, p=0.01, respectively). In addition, 13 (54%) of 24 patients in the acetaminophen groups combined exceeded the upper limit of their therapeutic range by 0.3 or greater compared with only 2 (17%) of 12 patients in the placebo group. No statistically significant differences in serum ALT or AST levels between either acetaminophen group versus the placebo group were found at week 4; however, there was a statistically significant increase in mean ALT level at week 2 in the acetaminophen 4-g/day group versus the placebo group. CONCLUSION: These findings support the existence of a clinically significant interaction between warfarin and daily use of acetaminophen 2-4 g, necessitating close monitoring of patients who receive this drug combination. Whether this interaction occurs when acetaminophen is taken in lower doses or is used sporadically requires further study. PMID- 17461703 TI - Rates of hospitalizations and emergency department visits in patients with asthma and chronic obstructive pulmonary disease taking beta-blockers. AB - STUDY OBJECTIVE: To determine the rates of hospitalizations and emergency department (ED) visits during cardioselective and nonselective beta-blocker therapy in patients with asthma and/or chronic obstructive pulmonary disease (COPD). DESIGN: Retrospective, observational cohort study. DATA SOURCE: Electronic medical records database. PATIENTS: A total of 11,592 adult patients with asthma and/or COPD, identified from August 1, 1997-December 31, 2005, who were taking beta-blockers for at least 30 days or had never received a beta blocker (controls). MEASUREMENTS AND MAIN RESULTS: Of these patients, 3062 were taking cardioselective and 690 nonselective beta-blockers; 7840 were controls. The primary end point for the beta-blocker groups was the rate of hospitalizations and ED visits/patient-year of beta-blocker therapy relative to the control group. In patients with asthma with or without concomitant COPD, cardioselective beta-blockers were associated with a relative risk of 0.89 (95% confidence interval [CI] 0.53-1.50) for hospitalizations and 1.40 (95% CI 1.20 1.62) for ED visits compared with controls. Nonselective beta-blockers were associated with a relative risk of 2.47 (95% CI 1.37-4.48) for hospitalizations and 1.21 (95% CI 0.91-1.62) for ED visits. In patients with COPD only, cardioselective beta-blockers were associated with a relative risk of 0.64 (95% CI 0.43-0.96) for hospitalizations and 1.19 (95% CI 1.02-1.39) for ED visits. Nonselective beta-blockers were associated with a relative risk of 1.02 (95% CI 0.52-2.02) for hospitalizations and 0.51 (95% CI 0.33-0.80) for ED visits. CONCLUSION: In patients with asthma with or without COPD, both cardioselective and nonselective beta-blocker use increased hospitalizations and ED visits compared with controls. Thus, these patients should receive beta-blocker therapy only if their cardiac risk exceeds their pulmonary risk and if they have concomitant cardiac disease for which beta-blockers decrease mortality, such as previous acute myocardial infarction or chronic heart failure. In patients with COPD only, cardioselective beta-blockers slightly increased the risk of ED visits but reduced the risk of hospitalizations. Nonselective beta-blocker therapy in these patients reduced the rate of ED visits and total visits. These findings suggest a larger safety margin with beta-blocker therapy in patients with COPD only than in those with asthma with or without COPD. PMID- 17461704 TI - Risk of major bleeding with concomitant dual antiplatelet therapy after percutaneous coronary intervention in patients receiving long-term warfarin therapy. AB - STUDY OBJECTIVES: To characterize the safety of concomitant aspirin, clopidogrel, and warfarin therapy after percutaneous coronary intervention (PCI), and to identify patient characteristics that increase the risk of hemorrhage. DESIGN: Retrospective, matched cohort study. SETTING: Academic medical center and affiliated outpatient offices. PATIENTS: The active group consisted of 97 patients who underwent PCI from January 1, 2000-September 30, 2005, and received warfarin, aspirin, and clopidogrel; the control group consisted of 97 patients who were individually matched to patients in the active group by procedure type, procedure year, age, and sex. Control patients received aspirin and clopidogrel. MEASUREMENTS AND MAIN RESULTS: Clinical data were collected from inpatient records, outpatient physician office records, and telephone surveys administered to patients or caregivers. The primary end point was major bleeding. The median duration of follow-up after index procedure was 182 days (range 0-191 days) in the active group and 182 days (range 0-213 days) in the control group. Fifty seven (59%) of the 97 patients in the active group received warfarin for atrial fibrillation. There were 14 major bleeds in the active group (including 1 death) and 3 major bleeds in the control group during the study period. Mean international normalized ratio at the time of bleeding was 3.4. Hazard ratio for major bleeding was 5.0 in patients receiving warfarin therapy (95% confidence interval 1.4-17.8, p=0.012). Aspirin dose, age, sex, body mass index, history of hypertension, diabetes mellitus, intraprocedural glycoprotein IIb-IIIa or anticoagulant type, and postprocedural anticoagulant use did not have a significant effect on the risk of major bleeding. CONCLUSION: Warfarin was an independent predictor of major bleeding after PCI in patients receiving dual antiplatelet therapy. Prospective data to further characterize the safety of concomitant warfarin and dual antiplatelet therapy after PCI are needed. PMID- 17461705 TI - Tolerability of beta-blockers in outpatients with refractory heart failure who were receiving continuous milrinone. AB - STUDY OBJECTIVE: To investigate the dosing, tolerability, and outcomes associated with the use of concomitant beta-blockers and inotropic therapy in patients with refractory heart failure during the first 6 months of their therapy. DESIGN: Retrospective review. SETTING: University-based, tertiary care heart failure and transplant center. PATIENTS: Sixteen inotrope-dependent outpatients with end stage refractory heart failure who were receiving continuous intravenous milrinone. Of these patients, 12 also received an oral beta-blocker; the remaining four patients who did not receive beta-blockers served as the comparator group. MEASUREMENTS AND MAIN RESULTS: For each patient, the initial and final study drug doses of continuous intravenous milrinone and oral beta blocker treatment, when applicable, were recorded over the 6-month period. Mean heart rate, blood pressure, ejection fraction, and oxygen consumption were measured, and 95% confidence intervals were calculated. Serum sodium and creatinine concentrations, as well as the creatinine clearance, were measured. In the 12 patients who received concomitant milrinone and beta-blockers, the mean baseline ejection fraction was approximately 18%, and they received milrinone for 18.6 weeks. Seven patients received carvedilol for 16.1 weeks, and five received metoprolol tartrate for 17.6 weeks. Dosages of the beta-blockers were titrated. Final daily doses were carvedilol 42.8 mg (95% confidence interval 20.3-65.4) and metoprolol 42.5 mg (95% confidence interval 28.0-57.2). Patients continued to receive other standard oral drug therapy for heart failure. One patient discontinued metoprolol and one discontinued carvedilol because of hypotension and/or worsening heart failure. Cardiac adverse events in the concomitant milrinone plus beta-blocker group were heart failure requiring hospitalization in 10 patients and ventricular arrhythmias in one. CONCLUSION: Inotrope-dependent patients with refractory end-stage heart failure tolerated continuous intravenous milrinone plus beta-blockers in addition to diuretics and vasodilators for the 6 month observation period. Beta-blocker dosages were titrated, and three patients achieved the target beta-blocker dosage established for stage A-C heart failure. Additional studies are needed to determine the optimal selection and dosing of drug combinations in this population. PMID- 17461706 TI - Prevalence of anemia in clinic patients with heart failure and cost analysis of epoetin treatment. AB - STUDY OBJECTIVES: To determine the prevalence of anemia in an outpatient heart failure clinic, describe the type of anemia in patients treated there, and evaluate the potential costs associated with epoetin therapy in this cohort. DESIGN: Single-center, retrospective cohort analysis (part 1) and a literature based economic decision analysis (part 2). DATA SOURCE: Medical records from a multidisciplinary, outpatient, heart failure clinic, and published hospitalization and drug-use data. PATIENTS: We evaluated 170 adults with chronic heart failure who were enrolled in the clinic and for whom at least one complete blood count was recorded between January 1, 2003, and April 15, 2006. MEASUREMENTS AND MAIN RESULTS: In part 1, demographic and clinical data were extracted from electronic medical records. The overall prevalence of anemia was 47.6% or 47.1%, as based on World Health Organization or National Kidney Foundation definitions, respectively. Normocytic anemia was characterized in 75.0% of patients. In part 2, heart failure hospitalization rates and costs, drug acquisition, and drug administration were estimated by using the published literature. In a hypothetical cohort of 100 patients with heart failure and comorbid anemia, the costs associated with outpatient epoetin and intravenous iron therapy exceeded savings in hospitalization costs by $83,070. Results of 1 way sensitivity analyses generally confirmed robustness of the model. CONCLUSION: Anemia is a common comorbidity in patients with chronic heart failure treated in the outpatient clinic. Although the current evidence is insufficient to support the use of epoetin in this population, initial findings indicate that epoetin and intravenous iron therapy may be associated with positive clinical outcomes. From a pharmacoeconomic standpoint, however, a reduction in the cost of heart failure related hospitalization does not offset the cost of epoetin and intravenous iron therapy. PMID- 17461707 TI - Role of prescription omega-3 fatty acids in the treatment of hypertriglyceridemia. AB - A prescription form of omega-3 fatty acids has been approved by the United States Food and Drug Administration as an adjunct to diet for the treatment of very high triglyceride levels. The active ingredients of omega-3 fatty acids are eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), which are responsible for the triglyceride lowering. The prescription product contains a total of 0.84 g of these two active ingredients in every 1-g capsule of omega-3 fatty acids. The total EPA and DHA dose recommended for triglyceride lowering is approximately 2-4 g/day. Fish oil products containing EPA and DHA are available without a prescription, but the American Heart Association advises that therapy with EPA and DHA to lower very high triglyceride levels should be used only under a physician's care. In patients with triglyceride levels above 500 mg/dl, approximately 4 g/day of EPA and DHA reduces triglyceride levels 45% and very low density lipoprotein cholesterol levels by more than 50%. Low-density lipoprotein cholesterol levels may increase depending on the baseline triglyceride level, but the net effect of EPA and DHA therapy is a reduction in non-high-density lipoprotein cholesterol level. Alternatively, patients may receive one of the fibrates (gemfibrozil or fenofibrate) or niacin for triglyceride lowering if their triglyceride levels are higher than 500 mg/dl. In controlled trials, prescription omega-3 fatty acids were well tolerated, with a low rate of both adverse events and treatment-associated discontinuations. The availability of prescription omega-3 fatty acids, which ensures consistent quality and purity, should prove to be valuable for the medical management of hypertriglyceridemia. PMID- 17461708 TI - Pharmacist-managed vaccination program increased influenza vaccination rates in cardiovascular patients enrolled in a secondary prevention lipid clinic. AB - STUDY OBJECTIVES: To determine whether a vaccination program in a pharmacist managed secondary prevention lipid clinic increased influenza immunization rates in a high-risk population, and whether age or gender disparity existed among those vaccinated. DESIGN: Retrospective chart review. SETTING: Large, multispecialty, group practice. PATIENTS: A total of 476 and 266 patients seen at clinic visits during the 2003-2004 and 2004-2005 influenza seasons, respectively. MEASUREMENTS AND MAIN RESULTS: Immunization rates were compared before (2003-2004 influenza season) and after (2004-2005 influenza season) the implementation of the influenza vaccination program; chi2 analysis was used for all statistical inferences. Vaccination rates increased significantly from 39% to 76% (p<0.0001) after program implementation. No before-after difference in rates was noted based on gender. Before implementation, patients younger than 65 years were less likely versus those aged 65 years or older to receive the influenza vaccine (29% vs 58%, p<0.0001). Age disparity in vaccination rates was eliminated after initiation of the program. CONCLUSION: The pharmacist-managed program increased influenza vaccination rates in high-risk patients with cardiovascular disease in advance of the newly published secondary prevention guidelines. Age-related differences in the vaccination rates were eliminated after program implementation. PMID- 17461709 TI - Reengineering clinical operations in a medical practice to optimize the management of anemia of chronic kidney disease. AB - OBJECTIVES: To describe the clinic design, clinical evaluations, and treatment approaches used in a multidisciplinary clinic for management of anemia of chronic kidney disease (CKD), and to evaluate several selected clinical outcomes associated with this approach to anemia management. SETTING: University affiliated, division of nephrology, outpatient multidisciplinary model CKD clinic headed by a clinical pharmacist. PATIENTS: One hundred sixty-six patients with anemia of CKD who were referred by nephrologists and primary care providers to the multidisciplinary clinic from March 1, 2002-July 31, 2004. MEASUREMENTS AND MAIN RESULTS: Patients received darbepoetin alfa dosed on an every-other-week basis. If patients were already receiving once-weekly recombinant human erythropoietin (r-HuEPO), the darbepoetin alfa dose was calculated by using the darbepoetin alfa package insert conversion table. If patients were naive to previous erythropoietic therapy, the darbepoetin alfa dose was either 60 microg or 0.7 microg/kg. The dose and frequency of darbepoetin alfa and oral iron supplements were adjusted to achieve the National Kidney Foundation Kidney Disease Outcomes Quality Initiative (K/DOQI) targets for hemoglobin levels and iron measures. The primary outcome analyzed was the proportion of patients with at least 30 days of treatment who achieved a target hemoglobin level of 11.0 g/dl or greater. Of 128 patients who received at least 30 days of treatment, 100 (78%) attained the hemoglobin level (mean +/- SD 11.7 +/- 7 g/dl). Ninety-nine of 128 patients were originally naive to erythropoietic therapy; 77 (78%) of these 99 patients achieved the hemoglobin target in a mean +/- SD of 7.9 +/- 7.5 weeks. These data contrast with the data of 29 patients seen in the year previous to the reengineered clinic process, whereby only 12 (41%) of these comparable patients reached hemoglobin target with r-HuEPO therapy. Of the 77 previously erythropoietic-naive patients, 82% were receiving darbepoetin alfa every other week, 14% every 3 weeks, and 4% every 4 weeks at the time the hemoglobin target was achieved. Oral iron administration significantly increased the chance of achieving the K/DOQI targets for hemoglobin and iron. CONCLUSION: Redefining roles and practices of members of a clinical practice and reengineering processes for anemia management were effective in achieving and maintaining target hemoglobin and iron levels. PMID- 17461710 TI - Iontophoretic drug delivery system: focus on fentanyl. AB - Fentanyl iontophoretic transdermal system (ITS) is a novel, patient-activated drug delivery device used for the management of acute postoperative pain in the hospital setting. This credit-card-sized device uses an imperceptible current of 170 milliampere to actively deliver a fentanyl hydrochloride 40-microg dose into the vasculature over a 10-minute interval. The unit is programmed to lock out further doses after either 80 doses or 24 hours, whichever is reached first. When comparing fentanyl ITS with intravenously administered fentanyl, serum concentrations differ significantly at 10 minutes after the initial dose is administered: 0.1 ng/ml for fentanyl ITS versus 0.7 ng/ml for intravenous fentanyl. Fentanyl ITS absorption increases in a time-dependent fashion over the first 10 hours of dosing. Other pharmacokinetic parameters of fentanyl ITS are comparable to those of intravenous fentanyl after 24 hours (maximum concentration 1.37 and 1.82 microg/ml, time to maximum concentration 0.65 and 0.58 hr, and area under the concentration-time curve at 23-24 hrs 1.23 and 1.34 microg x hr/ml for fentanyl ITS and intravenous fentanyl, respectively,). This new technology exhibited superior analgesia compared with placebo in two placebo-controlled studies that used time to exit as a primary end point. In addition, fentanyl ITS proved equivalent to patient-controlled analgesia with intravenous morphine. Although adverse effects were congruent with those expected from pure-agonist opioids, subjects assigned to the ITS group did experience a higher rate of mild, clinically nonsignificant erythema at the system placement site. Judicious monitoring for opioid-induced respiratory depression is recommended for fentanyl ITS, although this adverse effect has not been observed in clinical trials. Fentanyl ITS may provide another useful alternative in the management of acute postoperative pain. PMID- 17461711 TI - Recombinant activated factor VII treatment of retroperitoneal hematoma in a patient with renal failure receiving enoxaparin and clopidogrel. AB - Enoxaparin is a low-molecular-weight heparin that has pharmacokinetic and therapeutic advantages over unfractionated heparin in certain clinical conditions. However, its administration is not without risk. We describe the case of a 70-year-old woman with numerous medical problems who developed severe retroperitoneal bleeding after receiving several therapeutic doses of subcutaneous enoxaparin that inadvertently were not adjusted for her renal function until day 14 of therapy. She had severe bleeding with hemodynamic instability and required massive transfusions of blood products. Her bleeding could be controlled only by administration of four doses of recombinant activated factor VII (factor VIIa) in addition to embolization of the bleeding sites through angiographic microcoiling. The patient's hemodynamic status improved, and her hemoglobin level stabilized. This case report provides evidence of the clinical effectiveness of factor VIIa use as part of the management of refractory enoxaparin-induced retroperitoneal bleeding. However, further studies are needed to validate the dose-response relationship and further support the clinical utility of factor VIIa in this life-threatening situation. PMID- 17461712 TI - Severe anaphylactic reaction after repeated intermittent exposure to lepirudin. AB - Lepirudin, a recombinant DNA derivative of hirudin, is used to prevent thromboembolic complications caused by heparin-induced thrombocytopenia type II. Anaphylactic and anaphylactoid reactions have been reported with its use in patients both with and without known previous exposure to lepirudin. We describe the case of a 57-year-old woman who received five uneventful courses of lepirudin therapy before having a severe anaphylactic reaction during administration of the intravenous bolus dose that began her sixth course. The patient experienced cardiorespiratory arrest but recovered from the reaction. The decision to administer lepirudin to a patient who has previously received it should be reached with due consideration of the risk:benefit ratio and strategies to manage risk resulting from readministration. Risk factors for an anaphylactic reaction to lepirudin may include use of an initial bolus dose, intravenous rather than subcutaneous administration, length of any single course of therapy beyond 3 days, and repeat administration of lepirudin within 100 days. PMID- 17461713 TI - Leuconostoc species-associated endocarditis. AB - Leuconostoc species are gram-positive cocci belonging to the Streptococcaceae family. The species were generally regarded as nonpathogenic and of little importance in clinical microbiology until several occurrences of Leuconostoc infections were reported in the literature. Unlike many gram-positive bacteria, Leuconostoc species commonly demonstrate high-level resistance to vancomycin, with preserved sensitivity to most other antibacterial agents. We describe a 55 year-old man who developed endocarditis caused by Leuconostoc species sensitive to vancomycin. The patient received an aortic valve replacement and was treated with penicillin G and gentamicin; he experienced no further complications. PMID- 17461714 TI - Severe lactic acidosis associated with linezolid use in a patient with the mitochondrial DNA A2706G polymorphism. AB - Linezolid, an oxazolidinone antimicrobial, exerts its effect by binding to bacterial 23S ribosomal RNA, preventing the formation of the initiation complex. Its use is associated with reversible hyperlactatemia and lactic acidosis, and inhibition of mitochondrial protein synthesis may be the mechanism underlying this adverse effect. We describe a 35-year-old woman who developed severe lactic acidosis after she received linezolid for 35 days to treat a disseminated infection with Mycobacterium avium-intracellulare complex. This patient was found to have the mitochondrial DNA polymorphism A2706G, a variation previously suggested to predispose individuals to linezolid-associated lactic acidosis. In the future, increased understanding of the mitochondrial genome and its associated polymorphisms may allow us to identify patients at risk for adverse effects that were previously classified as idiosyncratic. PMID- 17461715 TI - Emphysematous cholecystitis in a patient with gastrointestinal stromal tumor treated with sunitinib. AB - A 50-year-old man had a metastatic gastrointestinal stromal tumor that was refractory to imatinib. He was prescribed a 6-week course of treatment with oral sunitinib 50 mg/day. During the fourth week of his first cycle of treatment with the drug, the patient developed acute-onset, right upper quadrant pain associated with nausea, vomiting, and fever; laboratory tests revealed leukocytosis and mild hyperbilirubinemia. He was diagnosed with acute emphysematous cholecystitis, which was treated with broad-spectrum antibiotics and percutaneous cholecystostomy. His symptoms resolved, and he successfully completed his course of therapy with sunitinib. Using the Naranjo adverse drug reaction probability scale, a score of 5 was derived, which indicates that the likelihood was probable that this adverse event was caused by sunitinib. PMID- 17461716 TI - Destructive osteomyelitis associated with early secondary syphilis in an HIV positive patient diagnosed by Treponema pallidum DNA polymerase chain reaction. AB - A 20-year old man who had sex with men (MSM) presented with destructive osteomyelitis of the sternal bone and diffuse maculopapular rash. During laboratory evaluation he was found to have secondary syphilis and HIV with viral load of 28,000 copies per milliliter and CD4 count of 251 cells per microliter. Surgical debridement and biopsy of the sternal bone was performed. The biopsy examination demonstrated bone necrosis with perivascular infiltration of plasma cells and lymphocytes and rare hystiocytes. No granulomatous lesions were identified and acid-fast, fungal, silver, and Gram's stains did not show any organism. All cultures were negative. Real-time polymerase chain reaction (PCR) using probes targeting a pathogen-specific and highly conserved TpN47 gene of Treponema pallidum was performed on the DNA, extracted from the biopsy specimen and T. pallidum amplicons were detected. Patient was initially treated empirically with vancomycin, piperacillin/tazobactam and intravenous aqueous penicillin G. After confirming the diagnosis he completed 2 weeks of intravenous aqueous penicillin G treatment with resolution of osteomyelitis confirmed at follow-up visit after 6 weeks. Osteomyelitis is a rarely described manifestation of secondary syphilis. To the best of our knowledge, this is the first case of using T. pallidum DNA PCR to confirm the diagnosis of syphilitic osteitis. We suggest that osteomyelitis may be an underrecognized problem in patients with secondary syphilis, especially in HIV-coinfected individuals and PCR seems to be a valuable method in confirming the diagnosis. PMID- 17461717 TI - Immune reconstitution syndrome in a patient with AIDS with paradoxically deteriorating brain tuberculoma. AB - A 54-year-old man with an underlying AIDS experienced fever and lethargy. Magnetic resonance imaging (MRI) showed multiple small ring-enhancement lesions over pons, basal ganglion, thalami, and bilateral cerebral hemisphere. Because of the concurrent pulmonary tuberculosis (TB), presumptive diagnosis of tuberculous meningitis and brain tuberculoma was made. The patient's condition clinically improved after a 3-month anti-TB treatment coupled with highly active antiretroviral therapy (HAART), and his CD4-T lymphocyte count was increased from 17 cells/mm(3) (HIV viral load, 294,000 copies per milliliter) to 153 cells/mm(3) (HIV viral load, 5930 copies per milliliter). However, the follow-up MRI disclosed disappearance of some old brain lesions and development of some new ones; some previously identified tuberculoma became smaller in size, while some other enlarger. Of note, ring-enhanced brain lesions were found over the left frontal lobe and left posterior fossa with perifocal edema and hyperintensity in diffusion weighted MRI indicating abscess formation. Steroid was added based on the presumed paradoxical reaction of brain tuberculoma. Complete resolution of brain lesions was found on MRI 9 months later. Tuberculoma should be considered in a patient with AIDS with numerous intracranial lesions if TB involving other site(s) is definitively diagnosed, especially when the patient is receiving prophylactic trimethoprim-sulfamethoxazole and/or serologically negative for toxoplasmosis. Our report demonstrated the peculiar phenomenon of paradoxical reaction of brain tuberculoma during immune reconstitution and strengthens the belief that additional use of steroids for paradoxical reaction of brain tuberculoma is indicated after exclusion of other causes for the progressively enlarging brain lesions. PMID- 17461718 TI - Tenofovir and abacavir combination therapy: lessons learned from an urban clinic population. AB - Regimens containing abacavir (ABC), tenofovir (TDF), and lamivudine (3TC) have recently been demonstrated to have high failure rates. This poses a clinical dilemma of how to manage patients currently being treated with other regimens containing tenofovir/abacavir. We evaluated the outcomes of tenofovir/abacavir regimens in our clinical practice through a retrospective review of 2655 charts. Two hundred patients (7%) were on a tenofovir/abacavir-containing regimen. Fifty nine patients met the criteria for analysis and were grouped into three groups: (1) antiretroviral naive, (2) virally suppressed patients switched to TDF/ABC, and (3) patients with failure of their first antiretroviral regimen. Rates of viral suppression in the naive, switch, and first-failure groups were 95%, 86%, and 46%, respectively. In the first-failure group, viral suppression was 66% without and 18% with a preexisting M184V. A composite analysis of the groups revealed a success rate of 86% when the regimen contained zidovudine (ZDV) and 62% when it did not. No K65R mutations were noted. These findings support continued caution in the use of TDF/ABC in combination. However, these data suggest that this combination may be successfully used in selected situations such as in combination with ZDV. In patients already virally suppressed on a TDF/ABC-containing regimen, considerations include continuing the regimen or adding zidovudine, in the attempt to protect against the development of a K65R mutation and/or virologic failure, versus changing a stable regimen. PMID- 17461719 TI - Lack of effect of gastric acid-reducing agents on the pharmacokinetics of lopinavir/ritonavir in HIV-infected patients. AB - Recent studies have shown that coadministration of certain protease inhibitors (PIs) with gastric acid-reducing agents results in decreased plasma concentrations of the PI. To assess the effect of acid-reducing agents on lopinavir/ritonavir, data from two clinical trials (n = 38 and 190) were pooled. Both trials randomized antiretroviral-naive, HIV-infected patients to receive lopinavir/ritonavir 400/100 mg twice-daily or 800/200 mg once-daily in combination with stavudine and lamivudine, or tenofovir and emtricitabine. Concurrent administration of gastric acid-reducing agents including antacids of various brand names, proton pump inhibitors (omeprazole, esomeprazole, lansoprazole, pantoprazole, and rabeprazole), and H(2)-receptor antagonists (ranitidine, famotidine, cimetidine, and nizatidine) was reported in both trials. Lopinavir and ritonavir pharmacokinetic parameters were evaluated. Thirty subjects were considered users of acid-reducing agents at the times of pharmacokinetic evaluation. HIV-infected patients who received gastric acid reducing agents during administration of lopinavir/ritonavir-based treatment regimens did not appear to have a reduction in lopinavir or ritonavir exposures. PMID- 17461720 TI - An evaluation of awareness: attitudes and beliefs of pregnant Nigerian women toward voluntary counseling and testing for HIV. AB - The rate of HIV seroprevalence in Nigeria is troublesome because it is one of the highest prevalence rates worldwide. As in most developing countries, vertical transmission from mother to child accounts for most of the HIV infections in Nigerian children. The purpose of this study was to determine the awareness, attitudes, and beliefs of pregnant Nigerian women toward voluntary counseling and testing (VCT) for HIV. Two hundred forty pregnant women in Awka, Nigeria, completed questionnaires aimed at determining their willingness to accept or reject VCT. Furthermore, participants where questioned about their knowledge of HIV infection, routes of transmission, and treatment options. The majority of the women (87%) approved of VCT; of those who approved, 93% were aware that VCT could reduce the risk of transmission of HIV to their babies. All respondents who accepted VCT were willing to be tested if results remained confidential and 89% would accept if they were tested simultaneously with their partners. 69% of the women who refused VCT attribute their refusal to the social and cultural stigmatization associated with HIV. Overall, the acceptance of VCT appears to depend on the understanding that VCT has proven benefits for the unborn child. Sociocultural factors such as stigmatization of HIV-infected individuals appears to be the major barrier toward widespread acceptance of VCT in Nigeria, thus the development of innovative health education strategies is essential for providing women with information regarding the benefits of VCT and other means of prevention of mother-to-child transmission of HIV (PMTCT). PMID- 17461721 TI - Desire for a child among women living with HIV/AIDS in northeast Brazil. AB - In Brazil, an increasing proportion of new HIV infections and AIDS cases involve women of reproductive age. To describe the reproductive desire of women with HIV/AIDS and to identify factors associated with the desire for motherhood, a cross-sectional study was carried out in the referral hospital for infectious diseases in Ceara State, northeast Brazil. In total, 229 women were included in data analysis. Median age was 32 years (interquartile range, 26-37), and 63% had a monthly family income of less than 210 USD. Forty-nine percent were using a contraceptive method, and 37% wished to undergo tubal ligation. Sixty-four percent of the latter women were motivated by the fear of having an HIV-positive child. Forty percent of the participants wanted to have a child. In the multivariate regression analysis, variables independently associated with women's desire to have a child were: younger age (in years, odds ration [OR] = 0.94; 95% confidence interval [CI]: 0.90-0.98), number of children (OR = 0.73; 95% CI: 0.57 0.96), and partner's desire for a child (OR = 3.35; 95%CI: 1.75-6.39). Having a partner who did not know about the woman's positive serostatus was negatively associated with the woman's desire for a child (OR = 0.17; 95% CI: 0.04-0.69). No variable related to clinical status was significantly associated with the outcome variable. Our data showed that many unsterilized HIV-positive women in northeast Brazil, at whatever stage of illness, have a desire for children. We recommend that nondirective counseling, consisting of helping women evaluate their own feelings, goals and needs with respect to reproductive options be provided. PMID- 17461722 TI - Desire to have children: gender and reproductive rights of men and women living with HIV: a challenge to health care in Brazil. AB - Links between HIV/AIDS care and reproductive health, including fertility options for people living with HIV (PLWH), have not been sufficiently addressed by health care providers. Moreover, few studies have addressed men in this regard. To describe attitudes toward parenthood and identify factors associated with desire to have children among men and women living with HIV a cross-sectional study involving a sample of 533 women and 206 men (bisexual and heterosexual) attending two reference sexually transmitted disease (STD)/AIDS centers in Sao Paulo, Brazil. Participants answered a standardized questionnaire. Desire to have children as the study outcome was compared between men and women and associated factors searched for in multivariable regression analysis. In contrast to previous studies conducted in developed countries, desire to have children in this sample was more frequent among men than among women and it was reported by 27.9% of participants (50.1% of men versus 19.2% of women). Women were more likely to anticipate doctors' strong opposition to PLWH getting pregnant and men reported lower information level about HIV/mother-to-child transmission (MTCT). Bisexual men were more likely to desire to have biologic children. Male gender, younger age, having no children, living with 1-2 children, and being in a heterosexual partnership were independently associated with desire to have children. Regardless of gender, the childless as well as the youngest should be regarded as groups to be particularly targeted by counseling, to be provided with objective information about reproductive rights and options. Further research is warranted to address the desire for children among strictly homosexual men. PMID- 17461723 TI - Prevalence and determinants of fertility intentions of HIV-infected women and men receiving antiretroviral therapy in South Africa. AB - Despite the increased emphasis on antiretroviral therapy (ART) and other health care services for HIV-infected individuals in sub-Saharan Africa, issues of fertility and childbearing have received relatively little attention. In particular, little is known about the prevalence and determinants of fertility intentions among HIV-infected women and men who are receiving ART. We conducted a cross-sectional study from August to November 2005 investigating these issues among patients attending a public sector ART service who had been receiving ART for at least one month. Overall, 311 individuals were interviewed (median age, 33 years) and 29% (n = 89) stated that they wanted to have children in the future. This proportion was slightly higher among males than females (36% versus 26%, p = 0.09). In a multivariate model predicting fertility desire among all participants, fertility desire was associated with male gender (odds ratio (OR):2.58; 95% confidence interval [CI]:1.29-5.08), younger age (OR: 0.92; 95% CI: 0.87-0.97), decreased number of children (OR: 0.32; 95% CI: 0.15-0.69), and being in a relationship of less than 5 years (OR: 3.93; 95% CI: 1.91-8.08). In addition, fertility desire was associated with increasing duration of ART among female participants, but not among males. These results suggest that a substantial proportion of HIV-infected women and men receiving ART in this setting would like to have children in the future. This highlights the importance of incorporating fertility-related counseling, as well as contraception and advice regarding safe conception and childbirth, as appropriate, into HIV treatment services. These findings also suggest that fertility desires may change through time and thus require ongoing attention as part of long-term care. PMID- 17461724 TI - Generic DDI oral solution approved. PMID- 17461725 TI - Hepatitis declining. PMID- 17461726 TI - Preventing and treating lymphatic minute metastasis with (32)p-chromic phosphate during an operation. AB - AIMS: The aim of this work was to study the effectiveness of (32)P-chromic phosphate colloids ((32)P-CP), by stromal injection during cardiac-esophageal carcinoma resection, in the prevention and treatment of minute lymphatic metastasis. METHODS: A multipointed infiltrative injection was done on 91 patients with clinically diagnosed cardiac-esophageal carcinoma to the adipose connective tissues of the corresponding bed of the resected lesion or retroperitonial wall, or tissues neighboring the left gastric artery, abdominal aorta, fenestra of aortic arch, pulmonary hilum, and superior mediastinum following the resection of a tumor. For patients with nonresectable lesions, a multipointed superficial injection of (32)P-CP-diluted solution was administered to the tumor and its surrounding invaded tissue, gastric artery, and the mediastinal adipose connective tissue. The dosage of (32)P-CP was 296 ~ 370 MBq/10 mL (8 ~ 10 mCi/10 mL). A simple operative group of 99 cases with clinically diagnosed cardiac-esophageal carcinoma, but without being given an injection of (32) P-CP, served as the control. Survival curves between the 2 groups were obtained using the Kaplan-Meier method and were compared by a log rank test. The differences in complications following the operation, the positive rate of lymph nodes (LN), the rate of LN metastasis, and the survival rate following the operation between the 2 groups were determined by conducting a monofactorial analysis by a chi(2) test. RESULTS: No operative deaths occurred in either group. The incidences of perioperative complications were not different statistically between the 2 groups (p > 0.05). The rate of lymph node metastasis, as well as the incidences of mediastinum, supraclavicle, and abdominal lymph node metastasis, showed that there were prominent statistical differences between the 2 groups (p < 0.05). A Kaplan-Meier analysis showed that there was a significant difference between the operative plus irradiation group, compared with the simple operative group (p = 0.0085). CONCLUSIONS: Stromal medication with (32)P-CP during a cardiac-esophageal carcinoma resection is a simple and safe procedure for controlling postoperative lymphatic metastasis and revealed a very fair clinical effectiveness in improving both medium- and long-term survival rates in patients. PMID- 17461727 TI - A novel pretargeting method for measuring antibody internalization in tumor cells. AB - A novel pretargeting method has been developed to quantitate antibody cellular internalization. In this study, the antibody was conjugated with a phosphorodiamidate morpholino oligomer (MORF) specific for the complementary MORF (cMORF) as an effector. Half the tumor cells were incubated with the MORF antibody (pretargeting group) and the other half with the same MORF-antibody at the same concentration but radiolabeled (direct targeting group). After incubation, the same dosage of radiolabeled cMORF was added to the wells of the pretargeting group. The radioactivity of the direct targeting cells represented the sum of both internalized and cell-surface-bound antibodies, whereas the radioactivity of the pretargeting cells resulted only from the surface-bound antibodies, as the radiolabeled cMORF does not penetrate the cell surface. Therefore, the difference in radioactivity accumulation between pretargeting and direct targeting provides the internalized fraction. In this example, the internalization of a MORF conjugated anti-prostate-specific membrane antigen antibody, 3C6, in LNCaP cells was examined, and the average cell-surface residence time was determined as 2 hours. This method of measuring antibody internalization is directly applicable to pretargeting applications but can be a universal alternative to the conventional acid-wash method, with the advantage of leaving the cell membrane undamaged. PMID- 17461728 TI - Implementing dosimetry in GATE: dose-point kernel validation with GEANT4 4.8.1. AB - GATE is a recent Monte Carlo code, based on GEANT4, and used in nuclear medicine mainly for imaging and detector design. Our goal was to implement dosimetry within GATE (i.e., combining the excellent potential of Gate for image modeling with GEANT4 dosimetric capabilities. The latest release of GEANT4 (4.8.1) completely revised the electron multiple scattering propagation algorithm. In this work, we calculated dose point kernels (DPK) for 0.01, 0.05, 0.1, 1, and 3 MeV monoenergetic electrons. We then compared our results with data obtained with another Monte Carlo code (MCNPX) or from the reference publication from Berger and Seltzer. To facilitate comparison, all calculated dose distributions were scaled to the corresponding R(CSDA), as given by the ESTAR NIST web database. Some GEANT4 parameters (i.e., Stepmax), or the shell thickness, had to be adjusted in order to achieve good agreement for energies below 1 MeV. For all energies except 10 keV, calculated DPKs do not differ significantly from the reference, as assessed by a Kolmogorov-Smirnov test. This preliminary step allowed us to consider the integration of GEANT4 dosimetric capabilities within the Gate framework. PMID- 17461729 TI - Current state of imaging protein-protein interactions in vivo with genetically encoded reporters. AB - Signaling pathways regulating proliferation, differentiation, and inflammation are commonly mediated through protein-protein interactions as well as reversible modification (e.g., phosphorylation) of proteins. To facilitate the study of regulated protein-protein interactions in cells and living animals, new imaging tools, many based on optical signals and capable of quantifying protein interactions in vivo, have advanced the study of induced protein interactions and their modification, as well as accelerated the rate of acquisition of these data. In particular, use of protein fragment complementation as a reporter strategy can accurately and rapidly dissect protein interactions with a variety of readouts, including absorbance, fluorescence, and bioluminescence. This review focuses on the development and validation of bioluminescent protein fragment complementation reporters that use either Renilla luciferase or firefly luciferase in vivo. Enhanced luciferase complementation provides a platform for near real-time detection and characterization of regulated and small-molecule-induced protein protein interactions in intact cells and living animals and enables a wide range of novel applications in drug discovery, chemical genetics, and proteomics research. PMID- 17461730 TI - Cell mechanics: integrating cell responses to mechanical stimuli. AB - Forces are increasingly recognized as major regulators of cell structure and function, and the mechanical properties of cells are essential to the mechanisms by which cells sense forces, transmit them to the cell interior or to other cells, and transduce them into chemical signals that impact a spectrum of cellular responses. Comparison of the mechanical properties of intact cells with those of the purified cytoskeletal biopolymers that are thought to dominate their elasticity reveal the extent to which the studies of purified systems can account for the mechanical properties of the much more heterogeneous and complex cell. This review summarizes selected aspects of current work on cell mechanics with an emphasis on the structures that are activated in cell-cell contacts, that regulate ion flow across the plasma membrane, and that may sense fluid flow that produces low levels of shear stress. PMID- 17461731 TI - Therapeutical potential of direct thrombin inhibitors for atherosclerotic vascular disease. AB - Adverse cardiovascular events are the consequence of a molecular chain reaction at the site of vulnerable plaques. Key players are platelets and coagulation factors that are activated following plaque rupture and often cause arterial obstruction. Thrombin, a plasma serine protease, plays a role in hemostasis of coagulation as well as in thrombosis and cell growth, leading to restenosis and atherosclerosis. Interesting and promising new molecules, the direct thrombin inhibitors, have been shown to be as effective as the combination of glycoprotein IIb-IIIa inhibitors and heparin for the prevention of arterial thrombosis. Until recently, direct thrombin inhibitors could be applied only parenterally; therefore, therapy was limited to hospitalized patients. As a result of recent drug development, orally active direct thrombin inhibitors are now available and have been evaluated for the long-term treatment of venous thrombosis and arterial fibrillation. Due to their specific pharmacodynamic characteristics by binding directly to thrombin--and thus inhibiting platelet aggregation and fibrin generation--these novel drugs may also have therapeutic potential for the treatment of atherothrombotic disease and its complications such as myocardial infarction, stroke or limb ischemia. PMID- 17461732 TI - Focus on acetylation: the role of histone deacetylase inhibitors in cancer therapy and beyond. AB - Reversal of tumorigenic epigenetic alterations is an exciting strategy for anticancer drug development. Pharmacologic inhibition of histone deacetylases (HDACs) induces differentiation, proliferation arrest and apoptosis of cancer cells. In addition to their effects on histones, HDAC inhibitors increase the acetylation level of several non-histone proteins, such as transcription factors, cytoskeletal proteins and molecular chaperones, which are crucial in tumorigenesis. Most importantly, the therapeutic potential of HDAC inhibitors goes well beyond carcinogenesis and may include neurodegenerative and inflammatory disorders. This editorial discusses the implication of HDACs in carcinogenesis, the molecular basis of the selectivity of HDAC inhibitors and their possible therapeutic role in non-malignant pathologic conditions. PMID- 17461733 TI - Aminoacyl-tRNA synthetases: essential and still promising targets for new anti infective agents. AB - The emergence of resistance to existing antibiotics demands the development of novel antimicrobial agents directed against novel targets. Historically, bacterial cell wall synthesis, protein, and DNA and RNA synthesis have been major targets of very successful classes of antibiotics such as beta-lactams, glycopeptides, macrolides, aminoglycosides, tetracyclines, rifampicins and quinolones. Recently, efforts have been made to develop novel agents against validated targets in these pathways but also against new, previously unexploited targets. The era of genomics has provided insights into novel targets in microbial pathogens. Among the less exploited--but still promising--targets is the family of 20 aminoacyl-tRNA synthetases (aaRSs), which are essential for protein synthesis. These targets have been validated in nature as aaRS inhibition has been shown as the specific mode of action for many natural antimicrobial agents synthesized by bacteria and fungi. Therefore, aaRSs have the potential to be targeted by novel agents either from synthetic or natural sources to yield specific and selective anti-infectives. Numerous high-throughput screening programs aimed at identifying aaRS inhibitors have been performed over the last 20 years. A large number of promising lead compounds have been identified but only a few agents have moved forward into clinical development. This review provides an update on the present strategies to develop novel aaRS inhibitors as anti-infective drugs. PMID- 17461734 TI - Potential therapeutic applications of the cyclotides and related cystine knot mini-proteins. AB - Cyclotides are naturally occurring mini-proteins that have a cyclic peptide backbone and a knotted arrangement of three disulfide bonds. They are remarkably stable and have a diverse range of therapeutically useful biological activities, including antimicrobial and anti-HIV activity, although their natural function appears to be as plant defence agents. Cyclotides are amenable to chemical synthesis and the potential exists to graft new bioactivities onto their cyclic cystine knot framework as a way of stabilising peptide drugs. Over the last few years, proof-of-concept that bioactive peptide epitopes can be grafted onto cyclotides and related cystine knot mini-proteins has been obtained. The cystine knot framework is tolerant to a wide range of residue substitutions and is showing great promise as a scaffold in drug design and protein engineering. PMID- 17461735 TI - Potential of non-antiarrhythmic drugs to provide an innovative upstream approach to the pharmacological prevention of sudden cardiac death. AB - Sudden cardiac death (SCD) is the most common cause of death and often occurs in low-risk patients. Present prevention strategies, mainly confined to high-risk subjects (proposed implantable cardioverter defibrillators recipients), have a limited effect on SCD burden in the general population. A relatively unexplored strategy for extending SCD prevention could imply targeting the early (upstream) processes of the complex cascade leading to SCD by non-antiarrhythmic drugs (i.e., beta-blockers, aldosterone antagonists, angiotensin-converting enzyme inhibitors, angiotensin receptor-blocker agents, statins and omega-3 fatty acids). In this innovative pharmacological perspective, agents with upstream effects may also be used in high-risk patients in association with a strictly downstream intervention, such as the implantable cardioverter defibrillator, in an attempt to obtain an additive/synergetic effect. PMID- 17461736 TI - New treatments for acute humoral rejection of kidney allografts. AB - Acute antibody-mediated rejection (acute humoral rejection; AHR) of organ allografts usually presents as severe dysfunction with a high risk of allograft loss. Peritubular capillary complement C4d deposition with renal dysfunction, associated with circulating donor-specific anti-human leukocyte antigen alloantibodies, is diagnostic of AHR in kidney allografts. Removal of alloantibodies with suppression of antibody production and rejection reversal is now possible. Therapeutic strategies that include combinations of plasmapheresis (or immunoadsorption), tacrolimus, mycophenolate mofetil and/or intravenous immunoglobulins, as well as rituximab or splenectomy, have been recently used to successfully treat AHR. However, the optimal protocol to treat AHR still remains to be defined. Anti-CD20+ monoclonal antibody therapy (rituximab) aiming at depleting B cells and suppressing antibody production has been used as rescue therapy in some episodes of steroid- and antilymphocyte-resistant humoral rejection. Plasmapheresis and/or intravenous polyclonal immunoglobulin, as well as rituximab, have also been used to successfully desensitize selected high immunological risk patients in anticipation of a previously cross-match positive (or ABO incompatible) kidney transplantation. In the near future, the possible role of new specific anti-B-cell approaches or, possibly, of new anti-T-cell activation approaches using selective agents such as belatacept should be assessed to further refine the present treatment of humoral rejection. PMID- 17461737 TI - Voltage-gated Na+ channels in neuropathic pain. AB - Neuropathic pain occurs as a result of some form of injury to the nervous system. Although the basis of the disease remains to be fully elucidated, numerous studies have suggested a major role for ion channels in the pathogenesis of neuropathic pain. As Na+ channels play a fundamental role in not only the generation but also in the conduction of an action potential, they have received considerable attention in the aetiology of pain sensation and have become important pharmacological targets. In this review, the authors discuss the importance of specific Na+ channel isoforms in the pathophysiology of neuropathic pain and the present use of Na+ channel antagonists in the treatment of neuropathic pain. PMID- 17461738 TI - Investigational treatment for vascular cognitive impairment. AB - Cognitive impairment of any severity associated with cerebrovascular damage is defined as vascular cognitive impairment as proposed by O'Brien. This is a heterogeneous syndrome with many subtypes, the most prevalent being vascular cognitive impairment without dementia. Neuropathological studies confirm that cerebrovascular disease and Alzheimer's disease frequently coexist. Diagnosis depends on criteria for dementia and vascular pathologies. Brain imaging is an important diagnostic tool. Although there is no approved intervention specifically for vascular cognitive impairment, general treatments (such as antiplatelet and antihypertensives) aimed at the prevention and management of stroke are used. Evidence from randomised, placebo-controlled studies of cholinesterase inhibitors for vascular dementia suggests that there may be beneficial effects for cognitive function, and clinical global impression is more favourable for the cholinesterase inhibitors compared with placebo. The effect of memantine also seems to be modest and similar to the effect that is demonstrated in patients with Alzheimer's disease. The accumulated evidence is not as comprehensive as that which exists for Alzheimer's disease. The cholinesterase inhibitors and memantine have not been extensively studied in vascular cognitive impairment without dementia. For the purposes of this review, the authors focus on interventions that have been evaluated by randomised controlled trials. PMID- 17461739 TI - Histone deacetylase inhibitors in cancer therapy. AB - Histones are a family of nuclear proteins that interact with DNA, resulting in DNA being wrapped around a core of histone octamer within the nucleosome. Acetylation/deacetylation of histones is an important mechanism that regulates gene expression and chromatin remodeling. Histone deacetylase (HDAC) inhibitors are a new class of chemotherapeutic drugs that regulate gene expression by enhancing the acetylation of histones, and thus inducing chromatin relaxation and altering gene expression. HDAC inhibitors have been shown in preclinical studies to have potent anticancer activities. A range of structurally diverse HDAC inhibitors have been purified as natural products or synthetically produced. Due to the promising preclinical activity of these agents, numerous clinical trials have been initiated. In this review, the results of published data of single agent and combination trials of these drugs are reviewed, with a focus on dosing, scheduling and toxicity. Although still early in drug development, there is a picture that is starting to develop as to the common toxicities and which tumors seem to be the most susceptible to this class of drugs. PMID- 17461740 TI - Dasatinib for the treatment of Philadelphia chromosome-positive leukaemias. AB - BCR-ABL, a constitutively active tyrosine kinase, causes chronic myeloid leukaemia (CML). Rational development of drugs targeting BCR-ABL has significantly improved the treatment of CML. Imatinib (a BCR-ABL tyrosine kinase inhibitor) produces haematological and cytogenetic remissions across all phases of CML and is the present standard of care. Imatinib resistance occurs in a significant proportion of patients and mechanisms of resistance include BCR-ABL mutations and activation of alternate oncogenic pathways. Dasatinib is a novel, potent, multi-targeted oral kinase inhibitor. Preclinical and clinical investigations demonstrate that dasatinib effectively overcomes imatinib resistance and has further improved the treatment of CML. Dasatinib was recently approved by the FDA for use in Philadelphia-positive leukaemias in patients who are resistant or intolerant to imatinib. PMID- 17461741 TI - ANCA-associated vasculitis: new options beyond steroids and cytotoxic drugs. AB - Small vessel vasculitic syndromes--Wegener's granulomatosis, microscopic polyangiitis and renal limited vasculitis (which are associated with circulating antineutrophil cytoplasmic autoantibodies)--are an important cause of renal failure. Present immunosuppressive regimens that are based on cyclophosphamide have significantly increased survival rates. However, these treatments are toxic, increase the risk of infection and do not cure disease. Therefore, newer approaches are required. Understanding disease pathogenesis has allowed rational use for newer therapies such as rituximab, which depletes B cells. Unfortunately, blockade of promising targets such as TNF-alpha, which was thought to be a pivotal cytokine in inflammation, has not shown benefit in a randomised controlled trial. Better understanding of the pathogenesis of the disease is the key to the development of novel targeted therapies, which are urgently required to improve patient prognosis. Gene therapy with targeted delivery of specific proteins is an exciting future prospect. PMID- 17461742 TI - Tafenoquine: a promising new antimalarial agent. AB - Malaria remains an important cause of global morbidity and mortality. As antimalarial drug resistance escalates, new safe and effective medications are necessary to prevent and treat malarial infection. Tafenoquine is an 8 aminoquinoline antimalarial that is presently under development. It has a long half-life of approximately 14 days and is generally safe and well tolerated, although it cannot be used in pregnant women and individuals who are deficient in the enzyme glucose-6-phosphate dehydrogenase. In well-designed studies, tafenoquine was highly effective in both the radical cure of relapsing malaria and causal prophylaxis of Plasmodium vivax and P. falciparum infections with protective efficacies of > or = 90%. Given its causal activity and safety profile, tafenoquine represents a potentially exciting alternative to standard agents for the prevention and radical cure of malaria. PMID- 17461743 TI - Review of dalbavancin, a novel semisynthetic lipoglycopeptide. AB - Dalbavancin is a semisynthetic lipoglycopeptide that is derived from teicoplanin with an extended half-life that enables once-weekly dosing. It has potent in vitro activity against most gram-positive organisms, with lower minimum inhibitory concentration values than vancomycin and other investigational lipoglycopeptides. Dalbavancin is active against multi-drug-resistant pathogens such as methicillin-resistant Staphylococcus aureus and vancomycin-resistant enterococci, except for strains exhibiting vanA resistance. Several Phase II and III trials have established similar efficacy and safety of dalbavancin with comparator agents in the treatment of complicated skin and skin structure infections and in catheter-related bloodstream infections. Dalbavancin may serve as an appealing alternative agent in the treatment of gram-positive infections, especially with its convenient once-weekly regimen. PMID- 17461744 TI - Monotherapy with lopinavir/ritonavir. AB - Despite the unprecedented pace of development of drugs for the treatment of a viral disease and the unquestionable efficacy of antiretroviral therapy, there is a need for less toxic and cheaper regimens that could simplify the treatment of HIV infection without sacrificing efficacy. The favorable pharmacokinetic profile and the high genetic barrier of boosted protease inhibitors make them ideal candidates for use as monotherapy. Given the encouraging results of available studies on lopinavir/ritonavir monotherapy in patients with no prior failure with protease inhibitors, it may be warranted to conduct trials to investigate the cost-effectiveness of lopinavir/ritonavir monotherapy as second-line therapy in resource-constrained settings where virologic monitoring is not feasible. In addition, larger trials with longer follow up, with particular attention to the potential consequences of viral replication in sites where the penetration of protease inhibitors may be poor, are needed before this strategy can be considered for routine use. PMID- 17461745 TI - Tinidazole for the treatment of vaginal infections. AB - Tinidazole has been used for vaginal infection worldwide but not in the US for > 40 years. Recently, tinidazole has been re-introduced and approved by the FDA for trichomoniasis and restudied as an alternative to metronidazole for bacterial vaginosis. In vitro antimicrobial activity and pharmacokinetics studies indicate that tinidazole has minor but possibly relevant antimicrobial as well as pharmacokinetic advantages when compared directly with metronidazole. Clinical comparison has been infrequent although the limited head-to-head studies indicate minimal therapeutic advantage with tinidazole. Perhaps the more relevant differences relate to the enhanced tolerance and reduced toxicity of tinidazole. Ongoing, as yet incomplete, studies directly comparing the clinical efficacy of metronidazole and tinidazole for bacterial vaginosis should clarify the status of tinidazole; however, cure rates are unlikely to be significantly different. Although uncommon, high-level trichomonal metronidazole resistance can be reliably cured by using tinidazole, which is an invaluable advantage. PMID- 17461746 TI - Infliximab for the therapy of chronic sarcoidosis, Baughman RP, Drent M, Kavuru M et al.: Infliximab therapy in patients with chronic sarcoidosis and pulmonary involvement. Am. J. Respir. Crit. Care Med. (2006) 174(7):795-802. AB - Sarcoidosis is an inflammatory multiorgan disease in which the lungs are the most commonly affected. It can also involve the skin, lupus pernio being a common form of chronic cutaneous sarcoidosis. The histopathologically specific lesion is represented by non-caseating granulomas occurring in the involved organs, with TNF-alpha playing a role in granuloma generation. Several therapies are available, with corticosteroids representing the conventional therapy given as topic or systemic formulations. Anti-TNF-alpha therapies (such as etanercept or infliximab) have been assessed so far, the latter most commonly in refractory sarcoidosis. The discussed study evaluates the safety and efficacy of infliximab in chronic sarcoidosis with pulmonary manifestations. PMID- 17461747 TI - Health information processing from television: the role of health orientation. AB - The quintessential presence of television in modern American life has led to decades of research on the unhealthy effects of television. However, recent years have witnessed a surge in scholarship seeking to interrogate the positive health effects of television, particularly in the realm of incorporating health content into entertainment-based television programs. One of the important critical questions in the realm of the positive health effects of television focuses on the amount of health information learning contributed by health information content on television. This article takes a motivation-based approach to health information learning from television, arguing that health orientation influences the amount of health information learned by individuals from television. On the basis of 2 separate studies, the article demonstrates that individuals who learn health information from a variety of television programs are more health oriented than individuals who do not learn health information from these television programs. PMID- 17461748 TI - After 9/11: goal disruption, emotional support, and psychological health in a lower exposure sample. AB - The terrorist attacks of 9/11 were traumatic even for Americans who were not directly victimized or in the geographic vicinity. This study examined whether emotional support received through interaction with others buffered the association between terrorism-related stress and psychological health (depression, anxiety, and physical symptoms) for individuals with lower exposure to the attacks. Five hundred eleven college students from an eastern university completed measures of goal disruption (stress) from terrorism, received emotional support, and psychological health in the first 2 weeks of December 2001. The results indicate that received emotional support moderated the relationship between goal disruption and depression and somatic symptoms. These findings suggest that the provision of emotional support should be understood as a fundamental communication skill relevant to recovery from disaster events. PMID- 17461749 TI - A statewide evaluation of the effectiveness of media literacy training to prevent tobacco use among adolescents. AB - Researchers used a quasi-experiment (N = 723) conducted in the field and using both pretests and posttests to carry out a theory-based evaluation of the effectiveness of a media literacy curriculum implemented in Washington state. Results showed that reflective thinking concerning media message about tobacco increased for all media literacy participants, whether or not they had used tobacco previously. Changes in reflective thinking affected a range of decision making indicators. Lesson participants who had not used tobacco demonstrated greater change at earlier stages of decision making in ways that suggested a greater understanding of the persuasive techniques used by tobacco manufacturers, on indicators such as perceived realism, desirability, and similarity. Lesson participants who had tried tobacco demonstrated greater change at later stages of decision making on indicators such as perceived peer norms for tobacco use, identification with tobacco-related portrayals, and expectancies for tobacco use. All participants also showed increases in their ability and motivations to resist smoking-related influences. Overall, the results suggest that media literacy has important and somewhat different effects on those who have and those who have not experimented with tobacco use. The results also show the importance of measuring cognitive and affective indicators of decision making that may change gradually as participants gain experience putting lessons learned into action. PMID- 17461750 TI - SARS wars: an examination of the quantity and construction of health information in the news media. AB - The media have the power to sway public perception of health issues by choosing what to publish and the context in which to present information. The media may influence an individual's tendency to overestimate the risk of some health issues while underestimating the risk of others, ultimately influencing health choices. Although some research has been conducted to examine the number of articles on selected health topics, little research has examined how the messages are constructed. The purpose of this article is to describe an examination of the construction of news reports on health topics using aspects of the social amplification of risk model and the elaboration likelihood model of persuasion for theoretical direction. One hundred news media reports (print, radio, television, and Internet) were analyzed in terms of message repetition, context, source, and grammar. Results showed that health topics were more often discussed in terms of risk, by credible sources using strong language. This content analysis provides an empirical starting point for future research into how such health news may influence consumer's perceptions of health topics. PMID- 17461751 TI - Reservations of the spirit: the development of a culturally sensitive spiritual beliefs scale about organ donation. AB - This study investigated whether spiritual beliefs offered any explanation for why participants from Korea (N = 146), Japan (N = 134), and the United States (N = 146) were willing or reluctant to register as organ donors. A culturally appropriate measure of spiritual beliefs about organ donation, the Spiritual Beliefs Scale, was developed consisting of 2 factors: (a) Spiritual Connection and (b) Spiritual Concern. Spiritual Connection was a significant predictor of behavioral intention to become an organ donor for Korean respondents, whereas Spiritual Concern was a significant predictor of reluctance to become an organ donor for American respondents. Spiritual beliefs correlated as predicted with attitude toward organ donation and fear of bodily mutilation, showing that the Spiritual Beliefs Scale exhibited internal, external, and predictive validity. Across the 3-country sample, Spiritual Connection was associated with greater willingness to become an organ donor for women, whereas Spiritual Concern inhibited participation for men. Implications of these findings are discussed for developing culturally effective education and procurement campaigns. PMID- 17461752 TI - Social sides of health risks: stigma and collective efficacy. AB - Health threats may not occur in a vacuum; one may need others' support to address a given health condition. For example, in Namibia, parents dying from AIDS related illnesses leave their orphaned children in need of adoption. If people do not feel threatened by HIV personally, social threats might motivate them to action. We extend the extended parallel process model (Witte, 1992) to include 2 social perceptions: (a) stigma and (b) collective efficacy. We found that Namibian respondents (n = 400) who did not feel threatened by HIV personally showed a relationship between these social perceptions and their willingness to support those living with HIV and their willingness to adopt AIDS orphans. These effects appeared for those who did not assess HIV as a health threat, suggesting that social threats, combined with efficacy, may motivate intentions to adopt recommended actions. Practical applications and intervention designs are discussed. PMID- 17461753 TI - Individual differences and context: factors mediating recall of anti-drug public service announcements. AB - A sample of 597 participants was surveyed to examine factors that influence recall of antidrug public service announcements (PSAs). High sensation-seekers and polydrug users recalled somewhat more antidrug PSAs than low sensation seekers and nonusers. Regression analyses indicated that total hours of television viewing did not predict recall of televised antidrug PSAs; instead, recall was predicted by preference for specific program genres, such as sports, news shows, action programs, sitcoms, and stand-up/comedy channel programs, and individual-difference variables such as sensation-seeking, drug use, and gender. The findings provide more evidence for the sensation-seeking targeting (SENTAR) approach and demonstrate the relevance of selective exposure and program contexts in reaching potential drug users. PMID- 17461754 TI - Telemedicine: its effects on health communication. AB - This article analyzes telemedicine, the use of distant communication technologies within the context of clinical health care, and the effects it has on health communication. The main effect is that telemedicine has the capacity to substantially transform health care in both positive and negative ways and to radically modify personal face-to-face communication (Turner, 2003). This has tremendous implications for health communication scholars in that they can extend the telemedicine debate by integrating fresh insights into more acceptable approaches that will refine and humanize mediated channels of health communication. There are several key areas of telemedicine that need to be discussed (i.e., e-health services, clinical encounters, etc.), all of which are identified in this article. In addition to describing the past and current applications of telemedicine, this article provides a better understanding of unique needs, resources, problems, and opportunities germane to telemedicine services. PMID- 17461755 TI - Cultural sensitivity and health promotion: assessing breast cancer education pamphlets designed for African American women. AB - This study discusses the implications for cultural sensitivity of the rhetorical choices in breast cancer education materials developed specifically for African American audiences by national organizations. Using the PEN-3 model of cultural sensitivity as an analytic framework for a generative rhetorical criticism, this study revealed that adaptations have been made in some pamphlets to acknowledge African American cultural values related to community, self-reliance, spirituality, and distrust of the Western medical establishment, but many messages could be revised to achieve a more comprehensive, balanced, accurate, and audience-specific discussion of the breast cancer issue. Achieving cultural sensitivity in health promotion materials necessitates attention to nuanced meanings in messages, revision of questionable arguments and evidence, and avoidance of ambiguity. PMID- 17461758 TI - Simultaneous generation of aptamers to multiple gamma-carboxyglutamic acid proteins from a focused aptamer library using DeSELEX and convergent selection. AB - By using the in vitro selection method SELEX against the complex mixture of GLA proteins and utilizing methods to deconvolute the resulting ligands, we were able to successfully generate 2'-ribo purine, 2'-fluoro pyrimidine aptamers to various individual targets in the GLA protein proteome that ranged in concentration from 10 nM to 1.4 microM in plasma. Perhaps not unexpectedly, the majority of the aptamers isolated following SELEX bind the most abundant protein in the mixture, prothrombin (FII), with high affinity. We show that by deselecting the dominant prothrombin aptamer the selection can be redirected. By using this DeSELEX approach, we were able to shift the selection toward other sequences and to less abundant protein targets and obtained an aptamer to Factor IX (FIX). We also demonstrate that by using an RNA library that is focused around a proteome, purified protein targets can then be used to rapidly generate aptamers to the protein targets that are rare in the initial mixture such as Factor VII (FVII) and Factor X (FX). Moreover, for all four proteins targeted (FII, FVII, FIX, and FX), aptamers were identified that could inhibit the individual protein's activitity in coagulation assays. Thus, by applying the concepts of DeSELEX and focused library selection, aptamers specific for any protein in a particular proteome can theoretically be generated, even when the proteins in the mixture are present at very different concentrations. PMID- 17461759 TI - RNA aptamers targeting the carboxyl terminus of KRAS oncoprotein generated by an improved SELEX with isothermal RNA amplification. AB - Mutations in the KRAS gene occur frequently in various human tumors and are known to lead to malignant transformation. We isolated RNA aptamers targeting activated mutant KRAS proteins using an improved SELEX method by isothermal RNA amplification. RNA aptamers were selected against mutant KRAS (G12V) proteins, as well as a biotinylated 15-amino-acid peptide from the carboxyl terminal of KRAS that contains a farnesylation site. All the selected RNA aptamers bound to the basic carboxy-terminal region of KRAS protein and the highest K(D) value was 2.3 microM. By an in vitro scintillation proximity assay, we demonstrated that KRAS aptamers inhibited farnesylation moderately. From these aptamers, we determined a consensus sequence (U)CCAAGCAC(AC) that, when concatamerized, exhibited higher binding affinity to the carboxy-terminal region of KRAS protein. Further improvement of binding affinity between aptamers and KRAS protein might provide a new therapeutic approach for activated mutant KRAS proteins. PMID- 17461760 TI - Comparison between molecularly defined and conventional therapeutics in a conditional BCR-ABL cell culture model. AB - Accumulating knowledge about the molecular mechanisms causing human diseases can support the development of targeted therapies such as imatinib, a BCR-ABL specific tyrosine kinase inhibitor to treat chronic myeloid leukemia (CML). Here, we use lentivirus-mediated RNA interference (RNAi) targeting BCR-ABL and the downstream signaling molecules SHP2, STAT5, and Gab2 to compare the efficacy and specificity of molecularly defined therapeutics with that of conventional cytotoxic drugs (cytarabine, doxorubicin, etoposide) in a conditional BCR-ABL cell culture model. IC(50) values were determined for each drug in TonB cells cultured either with interleukin-3 (IL-3) or BCR-ABL, and molecularly defined therapies were studied using lentivirally expressed shRNAs. We demonstrate that conventional anti-leukemic drugs have small or no differential effects under different cell culture conditions, whereas both imatinib and specific RNAi significantly inhibit proliferation of TonB cells in the presence of BCR-ABL but not IL-3. To study molecularly defined combination therapy, we evaluated either imatinib in TonB cells with target-specific RNAi or we used lentiviral vectors to induce combinatorial RNAi through simultaneous expression of two shRNAs. These combination therapies result in increased efficacy without loss in specificity. Interestingly, combinatorial RNAi can specifically deplete TonB cell cultures in the presence of BCR-ABL, even without targeting the oncogene itself. This model provides a tool to evaluate potential therapeutic targets and to quantify efficacy and specificity preclinically of new combination therapies in BCR-ABL positive cells. PMID- 17461761 TI - Tolerance of RNA interference toward modifications of the 5' antisense phosphate of small interfering RNA. AB - Bringing RNA interference (RNAi) under the control of light will allow the spacing, timing, and degree of gene expression to be controlled. We have previously shown that RNAi by small interfering (si) RNA can be modulated through randomly incorporated photolabile groups. Our and others interest is to find key locations on siRNA that can completely block RNAi until irradiation releases completely active siRNA. Some literature suggests that the 5' phosphate of the antisense strand of siRNA cannot be modified without completely blocking RNAi. We have examined this site as a potential switch for light control of RNAi and present evidence that siRNA modified at the 5' antisense phosphate can still cause RNAi, although not at the level effected by fully native siRNA. This contrasts with results from the literature, which suggest that modification of the 5' antisense phosphate will completely abrogate RNAi in siRNA. We have used mass spectrometry to identify and quantitate possible impurities that may be responsible for residual RNAi and show that they are present at 1% or less. Our results suggest that there is an inherent tolerance of the RNAi machinery toward modification of the 5' antisense phosphate. PMID- 17461762 TI - Anti-coxsackieviral efficacy of RNA interference is highly dependent on genomic target selection and emergence of escape mutants. AB - Enteroviral diseases are widespread and impose significant importance in medicine. Although the outcome of diseases that are associated with enteroviruses such as myocarditis, pancreatitis, hepatitis, or encephalomyelitis might be fatal, no specific antiviral therapy is yet available. We and others have shown that RNA interference (RNAi) effectively limits picornaviral replication and cytopathogenicity and improves survival in susceptible mice. However, little is known about the dependence of short interfering RNA (siRNA) efficacy on target region selection and emergence of viral escape mutants that may limit the effect of RNAi. The results of our study indicate that antiviral siRNA should be targeted preferentially to nonstructural protein coding regions because siRNA efficacy was consistently found to be superior compared to noncoding or structural protein coding regions. Further more, emergence of viral escape mutants that harbor single point mutations in the central part of the siRNA binding motif are the major factor that limits early therapeutic siRNA efficacy. The appearance of viral escape mutants can be sufficiently suppressed by combined administration of at least three distinct siRNA molecules. Therefore, genomic target selection and viral escape mutants are the most critical factors that limit early RNAi directed against enteroviral genomes. Both obstacles can be circumvented by appropriate target selection and combined siRNA administration. PMID- 17461763 TI - Tricyclo-DNA containing oligonucleotides as steric block inhibitors of human immunodeficiency virus type 1 tat-dependent trans-activation and HIV-1 infectivity. AB - Replication of human immunodeficiency virus type 1 (HIV-1) is controlled by a variety of viral and host proteins. The viral protein Tat acts in concert with host cellular factors to stimulate transcriptional elongation from the viral long terminal repeat (LTR) through a specific interaction with a 59-residue stem-loop RNA known as the trans-activation responsive element (TAR). Inhibitors of Tat-TAR recognition are expected to block transcription and suppress HIV-1 replication. In previous studies, we showed that 2'-O-methyl (OMe) oligonucleotide mixmers containing locked nucleic acid (LNA) residues are powerful steric block inhibitors of Tat-dependent trans-activation in a HeLa cell reporter system. Here we compare OMe/LNA mixmer oligonucleotides with oligonucleotides containing tricyclo-DNAs and their mixmers with OMe residues in four different assays: (1) binding to the target TAR RNA, (2) Tat-dependent in vitro transcription from an HIV-1 DNA template directed by HeLa cell nuclear extract, (3) trans-activation inhibition in HeLa cells containing a stably integrated firefly luciferase reporter gene under HIV-1 LTR control, and (4) an anti-HIV beta-galactosidase reporter assay of viral infection. Although tricyclo-DNA oligonucleotides bound TAR RNA more weakly, they were as good as OMe/LNA oligonucleotides in suppressing in vitro transcription and trans-activation in HeLa cells when delivered by cationic lipid. No inhibition of in vitro transcription and trans-activation in HeLa cells was observed for tricyclo-DNA/OMe mixmers, even though their affinities to TAR RNA were strong and their cell distributions did not differ from oligonucleotides containing all or predominantly tricyclo-DNA residues. Tricyclo-DNA 16-mer showed sequence-specific inhibition of beta-galactosidase expression in an anti-HIV HeLa cell reporter assay. PMID- 17461764 TI - Specific apoptosis induction in human papillomavirus-positive cervical carcinoma cells by photodynamic antisense regulation. AB - Human papillomavirus type 18 (HPV18) is frequently detected in cervical cancer cells. The viral proteins E6 and E7 are expressed consistently and have oncogenic activities. The E7 protein binds to a tumor suppressor, the retinoblastoma gene product (pRB), however, leading to the stabilization of tumor suppressor, p53 protein. On the other hand, another viral product, E6, forms complexes with p53 and abrogates its function, resulting in tumor progression. These facts imply that the E6 oncogene is one of the ideal targets for directed gene therapy in HPV positive cervical cancer. In this study, we tried photodynamic antisense regulation of the antiapoptotic E6 expression using a photocross-linking reagent, 4,5',8-trimethylpsoralen, conjugated oligo(nucleoside phosphorothioate) (Ps-S Oligo). This photodynamic antisense strategy effectively elicited the apoptotic death of HPV18-positive cervical cancer cells through the selective repression of E6 mRNA and consequent stabilization of p53 protein. E7-mediated signals potentially activated the p53 function and mobilized the p53 pathway to deliver pro-apoptotic signals to the cancer cells, leading to the suppression of in vivo tumorigenesis. An extremely low concentration of cisplatin in addition to Ps-S Oligos further up-regulated p53 activity, provoking massive apoptotic induction. These results suggest that the photodynamic antisense strategy has the great therapeutic potential in HPV-positive cervical cancers. PMID- 17461765 TI - Self-assembling supramolecular complexes by single-stranded extension from plasmid DNA. AB - Self-assembling supramolecular complexes are of great interest for bottom-up research like nanotechnology. DNA is an inexpensive building block with sequence specific self-assembling capabilities through Watson-Crick and/or Hoogsteen base pairing and could be used for applications in surface chemistry, material science, nanomechanics, nanoelectronics, nanorobotics, and of course in biology. The starting point is usually single-stranded DNA, which is rather easily accessible for base pairing and duplex formation. When long stretches of double stranded DNA are desirable, serving either as genetic codes or electrical wires, bacterial expansion of plasmids is an inexpensive approach with scale-up properties. Here, we present a method for using double-stranded DNA of any sequence for generating simple structures, such as junctions and DNA lattices. It is known that supercoiled plasmids are strand-invaded by certain DNA analogs. Here we add to the complexity by using "Self-assembling UNiversal (SUN) anchors" formed by DNA analog oligonucleotides, synthesized with an extension, a "sticky end" that can be used for further base pairing with single-stranded DNA. We show here how the same set of SUN anchors can be utilized for gene therapy, plasmid purification, junction for lattices, and plasmid dimerization through Watson Crick base pairing. Using atomic force microscopy, it has been possible to characterize and quantify individual components of such supra-molecular complexes. PMID- 17461766 TI - Selective regression of cells expressing mouse cytoskeleton-associated protein 2 transcript by trans-splicing ribozyme. AB - Cytoskeleton-associated protein 2 (CKAP2) is known to be highly expressed in primary human cancers as well as most cancer cell lines. CKAP2 functions as microtubule stabilizer and probably as cell proliferation inducer, indicating that CKAP2 might be a potential anticancer target. In this study, we developed a specific ribozyme that can replace mouse CKAP2 (mCKAP2) RNA with new transcripts through trans-splicing reaction. This specific RNA replacement resulted in triggering of transgene activity selectively in mammalian cells that express the mCKAP2 RNA. Simultaneously, the ribozyme reduced the expression level of the target RNA in the cells. Noticeably, the ribozyme selectively induced activity of the suicide gene herpes simplex virus thymidine kinase in cells expressing the mCKAP2 RNA and thereby specifically retarded the survival of these cells with ganciclovir treatment. This mCKAP2-specific ribozyme will be useful for validation of the RNA replacement as cancer gene therapy approach in mouse model with syngeneic tumors. PMID- 17461767 TI - Effective anti-hepatitis B virus hammerhead ribozymes derived from multimeric precursors. AB - Endonucleolytic hammerhead ribozymes have advantages of inhibiting gene expression by acting specifically, independently of cellular pathways, and within all cell compartments. However, there are concerns about inefficient silencing because of reduced intracellular cleavage of target RNA by ribozymes. To enable production of defined single-unit ribozymes and thereby increase effectiveness, we developed self-cleaving multimeric cassettes that generate several trans acting ribozyme units from a single transcript. cis and trans ribozyme cleavage, as assessed in vitro against three different sites within the X sequence of hepatitis B virus (HBV), occurred efficiently and precisely according to predictions deduced from the ribozyme designs. Significant knockdown of markers of viral replication in transfected cultured liver-derived cells was achieved by multiribozyme Pol II expression cassettes. To assess silencing efficacy of RNA prepared in vitro, transcription and cis cleavage reactions were carried out to prepare defined single-unit ribozymes. Transfection of ribozyme RNA was capable of inhibiting HBV surface antigen secretion from liver-derived cells without associated elevation of interferon-alpha or interferon-beta secretion into the culture upernatants. The approach described here is potentially useful for several applications, such as generation of RNA interference (RNAi) effectors, which require rapid and inexpensive generation of defined RNA sequences. PMID- 17461768 TI - DNA-enzyme-mediated cleavage of human immunodeficiency virus type 1 Gag RNA is significantly augmented by antisense-DNA molecules targeted to hybridize close to the cleavage site. AB - DNA-enzymes (Dzs) usually cleave short synthetic target RNAs very efficiently, but this activity diminishes significantly when tested on full-length RNAs, primarily because of the rigid secondary structures near the target sequence. We identified two Dzs, one each for 81-17 and 10-23 Dz, which cleaved the human immunodeficiency virus type 1 (HIV-1) Gag RNA poorly. We sought to use short oligodeoxynucleotides (ODNs) with the hope that it will facilitate Dz-mediated cleavage. The efficiencies of several ODNs were analyzed for their ability to augment the 8-17 Dz-mediated cleavage. We observed that ODNs that hybridized close to 5' and 3' ends of the target sequence were able to enhance significantly 8-17 Dz-mediated cleavage activity in a dose-dependent manner. The same was true for 10-23 Dz with ODNs that hybridized close to the target site. Thus, it was possible to enhance significantly the cleavage activity of poorly cleaving HIV-1 Gag-specific Dzs by using sequence-specific ODNs. This combination of antisense and catalytic Dz will, in principle, result in more effective gene suppression that could be exploited for therapeutic purposes. PMID- 17461769 TI - In situ entry of oligonucleotides into brain cells can occur through a nucleic acid channel. AB - Brain tissue has become a challenging therapeutic target, in part because of failure of conventional treatments of brain tumors and a gradually increasing number of neurodegenerative diseases. Because antisense oligonucleotides are readily internalized by neuronal cells in culture, these compounds could possibly serve as novel therapeutic agents to meet such a challenge. In previous in vitro work using cell culture systems, we have demonstrated that intracellular delivery requires a vector such as cationic liposomes since free oligonucleotides remain largely trapped in the endocytic pathway following cellular uptake. Here we studied the cellular uptake properties of oligonucleotides by explants of rat brain (brain slices), and by in vivo brain tissue after administration of oligonucleotides by bolus injection. In contrast to in vitro uptake, we show that in brain slices oligonucleotides were taken up by neuronal and nonneuronal cells, irrespective of their assembly with cationic liposomes. In either case, a diffuse distribution of oligonucleotides was seen in the cytosol and/or nucleus. Uptake of oligonucleotides by brain slices as a result of membrane damage, potentially arising from the isolation procedure, could be excluded. Interestingly, internalization was inhibited following treatment of the tissue with antibody GN 2640, directed against a nucleic acid channel, present in rat kidney cells. Our data support the view that an analogous channel is present in brain tissue, allowing entry of free oligonucleotides but not plasmids. Indeed, for delivery of the latter and accomplishment of effective transfection, cationic lipids were needed for gene translocation into both brain slices and brain tissue in vivo. These data imply that for antisense therapy to become effective in brain, cationic lipid-mediated delivery will only be needed for specific cell targeting but not necessarily for delivery per se to accomplish nuclear deposition of oligonucleotides into brain cells and subsequent down-regulation of disease related targets. PMID- 17461770 TI - Nucleotide pyrophosphatase/phosphodiesterase 1 is responsible for degradation of antisense phosphorothioate oligonucleotides. AB - The rapid degradation of unmodified phosphodiester oligodeoxynucleotides (PO oligos) by exo -and endonucleases limits their application as antisense constructs and requires the synthesis and use of modified oligonucleotides. Phosphorothioate analogs of oligonucleotides (PS-oligos) are much more stable against nucleolytic degradation than their unmodified counterparts, and this is one of the reasons for which they are a promising class of antisense oligonucleotides. However, PS-oligos also undergo slow hydrolysis by enzymes present in plasma. The oligonucleotide degradation proceeds mainly from the 3' end, resulting in the formation of a typical ladder of shorter products and the release of the mononucleoside 5' -phosphorothioates. So far, little has been known concerning the molecular identity of the enzymes involved in the degradation of PS-oligos. We now identify the human plasma 3' -exonuclease responsible for their degradation as a soluble form of nucleotide pyrophosphatase/phosphodiesterase 1 (NPP1) (EC 3.1.4.1/EC 3.6.1.9), also known as the plasma cell differentiation antigen PC-1. We also show that adenosine or deoxyadenosine (alpha-thio)triphosphates can act as potent inhibitors of NPPs. PMID- 17461771 TI - The peptide nucleic acid targeted to a regulatory sequence of the translocated c myc oncogene in Burkitt's lymphoma lacks immunogenicity: follow-up characterization of PNAEmu-NLS. AB - The present study aims to evaluate the antigenicity of a PNA complementary to the Emu sequence (PNAEmu) with cancer therapeutic potential properties in Burkitt's lymphoma (BL). In BL cells, the c-myc oncogene is repositioned next to the Emu enhancer of the immunoglobulin (Ig) locus, due to chromosomal translocation, and up-regulated. PNAEmu linked to a nuclear localization signal peptide was shown specifically to block c-myc hyperexpression by inhibiting cell growth in vitro and in vivo. Recently, we reported that the administration of PNAEmu to mice, following inoculation with BL cells, hinders tumor growth without toxic effects. To investigate the potential use of PNAEmu in clinical applications further, we tested its antigenicity. Mice were inoculated with an emulsion of free PNA or PNA crosslinked to the immunogenic carrier keyhole limpet hemocyanin (KLH) with Freund's adjuvant. Antibodies to free PNA were undetected, whereas both IgG and IgM antibodies to PNA-KLH were detected in mouse serum 28 and 38 days after inoculation. PMID- 17461776 TI - Does the vessel wall partition oxygen away from the tissue? AB - The microvascular wall is metabolically active and plays a key role in maintaining homeostasis. Additionally, it regulates the delivery of nutrients to the tissue and removal of its byproducts. Large oxygen gradients have been found to occur across the vessel wall. By using pharmacologic challenges, studies have demonstrated that the vascular wall regulates oxygen release from the blood to the tissue. Thus, these findings lead to the hypothesis that vasoactive substances used clinically may inadvertently partition proportionately more oxygen to the vascular wall and reduce the amount received by the tissue, leaving it potentially at risk. PMID- 17461777 TI - Cdc34 C-terminal tail phosphorylation regulates Skp1/cullin/F-box (SCF)-mediated ubiquitination and cell cycle progression. AB - The ubiquitin-conjugating enzyme Cdc34 (cell division cycle 34) plays an essential role in promoting the G1-S-phase transition of the eukaryotic cell cycle and is phosphorylated in vivo. In the present study, we investigated if phosphorylation regulates Cdc34 function. We mapped the in vivo phosphorylation sites on budding yeast Cdc34 (yCdc34; Ser207 and Ser216) and human Cdc34 (hCdc34 Ser203, Ser222 and Ser231) to serine residues in the acidic tail domain, a region that is critical for Cdc34's cell cycle function. CK2 (protein kinase CK2) phosphorylates both yCdc34 and hCdc34 on these sites in vitro. CK2-mediated phosphorylation increased yCdc34 ubiquitination activity towards the yeast Saccharomyces cerevisiae Sic1 in vitro, when assayed in the presence of its cognate SCFCdc4 E3 ligase [where SCF is Skp1 (S-phase kinase-associated protein 1)/cullin/F-box]. Similarly, mutation of the yCdc34 phosphorylation sites to alanine, aspartate or glutamate residues altered Cdc34-SCFCdc4-mediated Sic1 ubiquitination activity. Similar results were obtained when yCdc34's ubiquitination activity was assayed in the absence of SCFCdc4, indicating that phosphorylation regulates the intrinsic catalytic activity of Cdc34. To evaluate the in vivo consequences of altered Cdc34 activity, wild-type yCdc34 and the phosphosite mutants were introduced into an S. cerevisiae cdc34 deletion strain and, following synchronization in G1-phase, progression through the cell cycle was monitored. Consistent with the increased ubiquitination activity in vitro, cells expressing the phosphosite mutants with higher catalytic activity exhibited accelerated cell cycle progression and Sic1 degradation. These studies demonstrate that CK2-mediated phosphorylation of Cdc34 on the acidic tail domain stimulates Cdc34-SCFCdc4 ubiquitination activity and cell cycle progression. PMID- 17461778 TI - Age-associated perturbations in glutathione synthesis in mouse liver. AB - The nature of the mechanisms underlying the age-related decline in glutathione (GSH) synthetic capacity is at present unclear. Steady-state kinetic parameters of mouse liver GCL (glutamate-cysteine ligase), the rate-limiting enzyme in GSH synthesis, and levels of hepatic GSH synthesis precursors from the trans sulfuration pathway, such as homocysteine, cystathionine and cysteine, were compared between young and old C57BL/6 mice (6- and 24-month-old respectively). There were no agerelated differences in GCL V(max), but the apparent K(m) for its substrates, cysteine and glutamate, was higher in the old mice compared with the young mice (approximately 800 compared with approximately 300 microM, and approximately 710 compared with 450 microM, P<0.05 for cysteine and glutamate in young and old mice respectively). Amounts of cysteine, cystathionine and Cys-Gly increased with age by 91, 24 and 28% respectively. Glutathione (GSH) levels remained unchanged with age, whereas GSSG content showed an 84% increase, suggesting a significant pro-oxidizing shift in the 2GSH/GSSG ratio. The amount of the toxic trans-sulfuration/glutathione biosynthetic pathway intermediate, homocysteine, was 154% higher (P<0.005) in the liver of old mice compared with young mice. The conversion of homocysteine into cystathionine, a rate-limiting step in trans-sulfuration catalysed by cystathionine beta-synthase, was comparatively less efficient in the old mice, as indicated by cystathionine/homocysteine ratios. Incubation of tissue homogenates with physiological concentrations of homocysteine caused an up to 4.4-fold increase in the apparent K(m) of GCL for its glutamate substrate, but had no effect on V(max). The results suggest that perturbation of the catalytic efficiency of GCL and accumulation of homocysteine from the trans-sulfuration pathway may adversely affect de novo GSH synthesis during aging. PMID- 17461779 TI - Identification of Protor as a novel Rictor-binding component of mTOR complex-2. AB - The mTOR (mammalian target of rapamycin) protein kinase is an important regulator of cell growth. Two complexes of mTOR have been identified: complex 1, consisting of mTOR-Raptor (regulatory associated protein of mTOR)-mLST8 (termed mTORC1), and complex 2, comprising mTOR-Rictor (rapamycininsensitive companion of mTOR)-mLST8 Sin1 (termed mTORC2). mTORC1 phosphorylates the p70 ribosomal S6K (S6 kinase) at its hydrophobic motif (Thr389), whereas mTORC2 phosphorylates PKB (protein kinase B) at its hydrophobic motif (Ser473). In the present study, we report that widely expressed isoforms of unstudied proteins termed Protor-1 (protein observed with Rictor-1) and Protor-2 interact with Rictor and are components of mTORC2. We demonstrate that immunoprecipitation of Protor-1 or Protor-2 results in the co immunoprecipitation of other mTORC2 subunits, but not Raptor, a specific component of mTORC1. We show that detergents such as Triton X-100 or n octylglucoside dissociate mTOR and mLST8 from a complex of Protor-1, Sin1 and Rictor. We also provide evidence that Rictor regulates the expression of Protor 1, and that Protor-1 is not required for the assembly of other mTORC2 subunits into a complex. Protor-1 is a novel Rictor-binding subunit of mTORC2, but further work is required to establish its role. PMID- 17461780 TI - The TUMOROUS SHOOT DEVELOPMENT2 gene of Arabidopsis encoding a putative methyltransferase is required for cell adhesion and co-ordinated plant development. AB - Mutations in the TUMOROUS SHOOT DEVELOPMENT2 (TSD2) gene reduce cell adhesion, and in strongly affected individuals cause non-coordinated shoot development that leads to disorganized tumor-like growth in vitro. tsd2 mutants showed increased activity of axial meristems, reduced root growth and enhanced de-etiolation. The expression domains of the shoot meristem marker genes KNAT1 and KNAT2 were enlarged in the mutant background. Soil-grown tsd2 mutants were dwarfed, but overall showed morphology similar to that of the wild-type (WT). The TSD2 gene was identified by map-based cloning. It encodes a novel 684 amino acid polypeptide containing a single membrane-spanning domain in the N-terminal part and S-adenosyl-l-methionine binding and methyltransferase domains in the C terminal part. Expression of a TSD2:GUS reporter gene was detected mainly in meristems and young tissues. A green fluorescent protein-tagged TSD2 protein localized to the Golgi apparatus. The cell-adhesion defects indicated altered pectin properties, and we hypothesize that TSD2 acts as a pectin methyltransferase. However, analyses of the cell-wall composition revealed no significant differences of the monosaccharide composition, the uronic acid content and the overall degree of pectin methylesterification between tsd2 and WT. The findings support a function of TSD2 as a methyltransferase, with an essential role in cell adhesion and coordinated plant development. PMID- 17461781 TI - Seed dormancy release in Arabidopsis Cvi by dry after-ripening, low temperature, nitrate and light shows common quantitative patterns of gene expression directed by environmentally specific sensing. AB - The depth of seed dormancy can be influenced by a number of different environmental signals, but whether a common mechanism underlies this apparently similar response has yet to be investigated. Full-genome microarrays were used for a global transcript analysis of Arabidopsis thaliana Cape Verde Island accession seeds exposed to dry after-ripening (AR), or low temperature, nitrate and light when imbibed. Germination studies showed that the sensitivity of imbibed seeds to low temperature, nitrate and light was dependent upon the length of time spent AR following harvest. Seeds had an absolute requirement for light to complete dormancy release in all conditions, but this effect required an exposure to a prior dormancy relieving environment. Principal component analyses of the expression patterns observed grouped physiological states in a way that related to the depth of seed dormancy, rather than the type of environmental exposure. Furthermore, opposite changes in transcript abundance of genes in sets associated with dormancy, or dormancy relief through AR, were also related to the depth of dormancy and common to different environments. Besides these common quantitative changes, environment-specific gene expression patterns during dormancy relief are also described. For example, higher transcript abundance for genes linked to the process of nitrate accumulation, and nitrate reduction was associated with dormancy relief. The quantity of GA3ox1 transcripts increased during dormancy relief in all conditions, in particular when dormancy relief was completed by exposure to light. This contrasts with transcripts linked to abscisic acid (ABA) synthesis, which declined. The results are consistent with a role for the ABA/gibberellic acid balance in integrating dormancy-relieving environmental signals. PMID- 17461782 TI - Deficiency of mitochondrial fumarase activity in tomato plants impairs photosynthesis via an effect on stomatal function. AB - Transgenic tomato (Solanum lycopersicum) plants expressing a fragment of a fumarate hydratase (fumarase) gene in the antisense orientation and exhibiting considerable reductions in the mitochondrial activity of this enzyme show impaired photosynthesis. The rate of the tricarboxylic acid cycle was reduced in the transformants relative to the other major pathways of carbohydrate oxidation and the plants were characterized by a restricted rate of dark respiration. However, biochemical analyses revealed relatively little alteration in leaf metabolism as a consequence of reducing the fumarase activity. That said, in comparison to wild-type plants, CO(2) assimilation was reduced by up to 50% under atmospheric conditions and plants were characterized by a reduced biomass on a whole plant basis. Analysis of further photosynthetic parameters revealed that there was little difference in pigment content in the transformants but that the rate of transpiration and stomatal conductance was markedly reduced. Analysis of the response of the rate of photosynthesis to variation in the concentration of CO(2) confirmed that this restriction was due to a deficiency in stomatal function. PMID- 17461783 TI - Members of the PHO1 gene family show limited functional redundancy in phosphate transfer to the shoot, and are regulated by phosphate deficiency via distinct pathways. AB - The PHO1 family comprises 11 members in Arabidopsis thaliana. In order to decipher the role of these genes in inorganic phosphate (Pi) transport and homeostasis, complementation of the pho1 mutant, deficient in loading Pi to the root xylem, was determined by the expression of the PHO1 homologous genes under the control of the PHO1 promoter. Only PHO1 and the homologue PHO1;H1 could complement pho1. The PHO1;H1 promoter was active in the vascular cylinder of roots and shoots. Expression of PHO1;H1 was very low in Pi-sufficient plants, but was strongly induced under Pi-deficient conditions. T-DNA knock-out mutants of PHO1;H1 neither showed growth defects nor alteration in Pi transport dynamics, or Pi content, compared with wild type. However, the double mutant pho1/pho1;h1 showed a strong reduction in growth and in the capacity to transfer Pi from the root to the shoot compared with pho1. Grafting experiments revealed that phenotypes associated with the pho1 and pho1/pho1;h1 mutants were linked to the lack of gene expression in the root. The increased expression of PHO1;H1 under Pi deficiency was largely controlled by the transcription factor PHR1 and was suppressed by the phosphate analogue phosphite, whereas the increase of PHO1 expression was independent of PHR1 and was not influenced by phosphite. Together, these data reveal that although transfer of Pi to the root xylem vessel is primarily mediated by PHO1, the homologue PHO1;H1 also contributes to Pi loading to the xylem, and that the two corresponding genes are regulated by Pi deficiency by distinct signal transduction pathways. PMID- 17461784 TI - ABA-Hypersensitive Germination1 encodes a protein phosphatase 2C, an essential component of abscisic acid signaling in Arabidopsis seed. AB - The phytohormone abscisic acid (ABA) regulates physiologically important stress and developmental responses in plants. To reveal the mechanism of response to ABA, we isolated several novel ABA-hypersensitive Arabidopsis thaliana mutants, named ahg (ABA-hypersensitive germination). ahg1-1 mutants showed hypersensitivity to ABA, NaCl, KCl, mannitol, glucose and sucrose during germination and post-germination growth, but did not display any significant phenotypes in adult plants. ahg1-1 seeds accumulated slightly more ABA before stratification and showed increased seed dormancy. Map-based cloning of AHG1 revealed that ahg1-1 has a nonsense mutation in a gene encoding a novel protein phosphatase 2C (PP2C). We previously showed that the ahg3-1 mutant has a point mutation in the AtPP2CA gene, which encodes another PP2C that has a major role in the ABA response in seeds (Yoshida et al., 2006b). The levels of AHG1 mRNA were higher in dry seeds and increased during late seed maturation--an expression pattern similar to that of ABI5. Transcriptome analysis revealed that, in ABA treated germinating seeds, many seed-specific genes and ABA-inducible genes were highly expressed in ahg1-1 and ahg3-1 mutants compared with the wild-type. Detailed analysis suggested differences between the functions of AHG1 and AHG3. Dozens of genes were expressed more strongly in the ahg1-1 mutant than in ahg3-1. Promoter-GUS analyses demonstrated both overlapping and distinct expression patterns in seed. In addition, the ahg1-1 ahg3-1 double mutant was more hypersensitive than either monogenic mutant. These results suggest that AHG1 has specific functions in seed development and germination, shared partly with AHG3. PMID- 17461785 TI - Purification of the M flax-rust resistance protein expressed in Pichia pastoris. AB - The M flax-rust resistance (R) gene is predicted to encode a 150-kDa protein of the Toll-interleukin-like receptor-nucleotide binding site-leucine rich repeat (TIR-NBS-LRR) class of plant disease resistance proteins and provides resistance against the Melampsora lini (flax rust) fungus carrying the AvrM avirulence gene. The extremely low level of this class of R proteins found in plant tissue has precluded their biochemical and structural analysis, and the study of these proteins has been largely restricted to genetic analyses and in vivo investigations. Here we report the production and purification of the M protein in the methalotrophic yeast, Pichia pastoris. Expression trials with five different constructs reveals optimum levels of soluble native M protein can be obtained as an N-terminally 9x His-tagged protein, in which the first 21 amino acids of the predicted wild-type protein are deleted. Expression was achieved using a high cell density fed-batch bioreactor culture at low temperature. M protein was purified to near homogeneity from whole-cell lysates using cation exchange, immobilised metal ion affinity chromatography and gel filtration with a final yield of approximately 3 mg of protein/1000 g wet weight of yeast cells lysed. The successful expression and purification of soluble M protein opens the way for biochemical and structural analysis of this class of important plant proteins. PMID- 17461786 TI - SHA1, a novel RING finger protein, functions in shoot apical meristem maintenance in Arabidopsis. AB - Post-embryonic plant growth is dependent on a functional shoot apical meristem (SAM) that provides cells for continuous development of new aerial organs. However, how the SAM is dynamically maintained during vegetative development remains largely unclear. We report here the characterization of a new SAM maintenance mutant, sha1-1 (shoot apical meristem arrest 1-1), that shows a primary SAM-deficient phenotype at the adult stage. The SHA1 gene encodes a novel RING finger protein, and is expressed most intensely in the shoot apex. We show that, in the sha1-1 mutant, the primary SAM develops normally during the juvenile vegetative stage, but cell layer structure becomes disorganized after entering the adult vegetative stage, resulting in a dysfunctional SAM that cannot initiate floral primordia. The sha1-1 SAM terminates completely at the stage when the wild type begins to bolt, producing adult plants with a primary inflorescence deficient phenotype. These observations indicate that SHA1, a putative E3 ligase, is required for post-embryonic SAM maintenance by controlling proper cellular organization. PMID- 17461787 TI - Immunogenic tagging of chloroplasts allows their isolation from defined cell types. AB - Chloroplast structure varies depending on cell type. Currently it is difficult to obtain insight into how differences in chloroplast structure relate to function, as it is often not possible to isolate chloroplasts from specific cells. To address this, we have developed an approach that involves labelling chloroplasts from individual cell types by placing the foreign yellow fluorescent protein (YFP) on their outer surface, and then isolating those labelled chloroplasts immunogenically. Cell specificity is achieved through the use of enhancer trap lines. When whole leaves are homogenized, a mixture of labelled and unlabelled chloroplasts is released, but magnetic beads coated with antibodies to the green fluorescent protein (GFP) allow the labelled chloroplasts to be isolated. Chloroplasts from spongy mesophyll, vascular and epidermal cells of Arabidopsis thaliana were obtained in this way, and semi-quantitative RT-PCR showed that the abundance of various chloroplast transcripts differed between these three cell types. The approach is based on genetic logic, and so could be applied to the isolation of various organelles or subcellular compartments from transformable organisms other than A. thaliana. PMID- 17461788 TI - Microcolinearity and genome evolution in the AdhA region of diploid and polyploid cotton (Gossypium). AB - Genome sizes vary by several orders of magnitude, driven by mechanisms such as illegitimate recombination and transposable element proliferation. Prior analysis of the CesA region in two cotton genomes that diverged 5-10 million years ago (Ma), and acquired a twofold difference in genome size, revealed extensive local conservation of genic and intergenic regions, with no evidence of the global genome size difference. The present study extends the comparison to include BAC sequences surrounding the gene encoding alcohol dehydrogenase A (AdhA) from four cotton genomes: the two co-resident genomes (A(T) and D(T)) of the allotetraploid, Gossypium hirsutum, as well as the model diploid progenitors, Gossypium arboreum (A) and Gossypium raimondii (D). In contrast to earlier work, evolution in the AdhA region reflects, in a microcosm, the overall difference in genome size, with a nearly twofold difference in aligned sequence length. Most size differences may be attributed to differential accumulation of retroelements during divergence of the genome diploids from their common ancestor, but in addition there has been a biased accumulation of small deletions, such that those in the smaller D genome are on average twice as large as those in the larger A genome. The data also provide evidence for the global phenomenon of 'genomic downsizing' in polyploids shortly after formation. This in part reflects a higher frequency of small deletions post-polyploidization, and increased illegitimate recombination. In conjunction with previous work, the data here confirm the conclusion that genome size evolution reflects many forces that collectively operate heterogeneously among genomic regions. PMID- 17461789 TI - Identification of a flavin-monooxygenase as the S-oxygenating enzyme in aliphatic glucosinolate biosynthesis in Arabidopsis. AB - The cancer-preventive activity of cruciferous vegetables is commonly attributed to isothiocyanates resulting from the breakdown of the natural products glucosinolates (GSLs). Sulforaphane, the isothiocyanate derived from 4 methylsulfinylbutyl GSL, is thought to be the major agent conferring cancer preventive properties, whereas the isothiocyanate of 4-methylthiobutyl GSL does not have the same activity. We report the identification of an Arabidopsis flavin monooxygenase (FMO) enzyme, FMO(GS-OX1), which catalyzes the conversion of methylthioalkyl GSLs into methylsulfinylalkyl GSLs. This is evidenced by biochemical characterization of the recombinant protein, and analyses of the GSL content in FMO(GS-OX1) overexpression lines and an FMO(GS-OX1) knock-out mutant of Arabidopsis. The FMO(GS-OX1) overexpression lines show almost complete conversion of methylthioalkyl into methylsulfinylalkyl GSLs, with an approximately fivefold increase in 4-methylsulfinylbutyl GSL in seeds. Identification of FMO(GS-OX1) provides a molecular tool for breeding of Brassica vegetable crops with increased levels of this important GSL, which has implications for production of functional foods enriched with the cancer preventive sulforaphane. PMID- 17461790 TI - Transcript and metabolite profiling during cold acclimation of Arabidopsis reveals an intricate relationship of cold-regulated gene expression with modifications in metabolite content. AB - Exposure of Arabidopsis to low temperatures results in cold acclimation where freezing tolerance is enhanced. To achieve a wider view of the role of transcriptome to biochemical changes that occur during cold acclimation, analyses of concurrent transcript and metabolite changes during cold acclimation was performed revealing the dynamics of selected gene-metabolite relationships. Exposure to low temperature resulted in broad transcriptional and metabolite responses. Principal component analysis revealed sequentially progressive, global changes in both gene expression and metabolite profiles during cold acclimation. Changes in transcript abundance for many metabolic processes, including protein amino acid biosynthetic pathways and soluble carbohydrates, during cold acclimation were observed. For some metabolic processes, changes in transcript abundance temporally correlated with changes in metabolite levels. For other metabolic processes, changes in transcript levels were not correlated with changes in metabolite levels. The present findings demonstrate that regulatory processes independent of transcript abundance represent a key part of the metabolic adjustments that occur during cold acclimation. PMID- 17461791 TI - The transcription factor HIG1/MYB51 regulates indolic glucosinolate biosynthesis in Arabidopsis thaliana. AB - Glucosinolates are a class of plant secondary metabolites that serve as antiherbivore compounds in plant defence. A previously identified Arabidopsis thaliana activation-tagged line, displaying altered levels of secondary metabolites, was shown here to be affected in the content of indolic and aliphatic glucosinolates. The observed chemotype was caused by activation of the R2R3-MYB transcription factor gene HIG1 (HIGH INDOLIC GLUCOSINOLATE 1, also referred to as MYB51). HIG1/MYB51 was shown to activate promoters of indolic glucosinolate biosynthetic genes leading to increased accumulation of indolic glucosinolates. The corresponding loss-of-function mutant hig1-1 contained low levels of glucosinolates. Overexpression of the related transcription factor ATR1/MYB34, which had previously been described as a regulator of indolic glucosinolate and indole-3-acetic acid homeostasis, in the hig1-1 mutant background led to a partial rescue of the mutant chemotype along with a severe high-auxin growth phenotype. Overexpression of MYB122, another close homologue of HIG1/MYB51, did not rescue the hig1-1 chemotype, but caused a high-auxin phenotype and increased levels of indolic glucosinolates in the wild-type. By contrast, overexpression of HIG1/MYB51 resulted in the specific accumulation of indolic glucosinolates without affecting auxin metabolism and plant morphology. Mechanical stimuli such as touch or wounding transiently induced the expression of HIG1/MYB51 but not of ATR1/MYB34, and HIG1/MYB51 overexpression reduced insect herbivory as revealed by dual-choice assays with the generalist lepidopteran herbivore, Spodoptera exigua. We hypothesize that HIG1/MYB51 is a regulator of indolic glucosinolate biosynthesis that also controls responses to biotic challenges. PMID- 17461792 TI - Arabidopsis Ovate Family Protein 1 is a transcriptional repressor that suppresses cell elongation. AB - Transcription factors regulate multiple aspects of plant growth and development. Here we report the identification and functional analysis of a plant-specific, novel transcription factor in Arabidopsis. We isolated a dominant, gain-of function mutant that displays reduced lengths in all aerial organs including hypocotyl, rosette leaf, cauline leaf, inflorescence stem, floral organs and silique. Molecular cloning revealed that these phenotypes are caused by elevated expression of the Arabidopsis thaliana Ovate Family Protein 1 (AtOFP1). This mutant was designated as Atofp1-1D. We show that the altered morphology of Atofp1 1D mutant is caused by reduced cell length resulting from reduced cell elongation, and demonstrate that a mutant harboring a transposon insertion that disrupts the OVATE domain of AtOFP1 is indistinguishable from wild-type plants. Plants overexpressing other closely related AtOFP genes phenocopy plants overexpressing AtOFP1, implying a possible overlapping function among members of the AtOFP gene family. We found that AtOFP1 localizes in the nucleus, and that AtOFP1 functions as an active transcriptional repressor. Chromatin immunoprecipitation results indicated that AtGA20ox1, a gene encoding the key enzyme in GA biosynthesis, is a target gene regulated by AtOFP1. Consistent with this, exogenous gibberellic acid can partially restore defects in cell elongation in plants overexpressing AtOFP1, suggesting that such a reduced cell elongation is caused, in part, by the deficiency in gibberellin biosynthesis. Taken together, our results indicate that AtOFP1 is an active transcriptional repressor that has a role in regulating cell elongation in plants. PMID- 17461793 TI - The KNOX gene SHOOT MERISTEMLESS is required for the development of reproductive meristematic tissues in Arabidopsis. AB - In Arabidopsis, the central stem cells of the shoot apical meristem (SAM) are indefinitely maintained, whereas those in floral meristems differentiate into female gametophyte-bearing organs termed carpels. The class 1 KNOX genes encode homeodomain transcription factors that function variously in the establishment and maintenance of the SAM, and have also been implicated in carpel development. Here we show that the KNOX gene SHOOT MERISTEMLESS (STM) induces carpel formation and promotes homeotic conversion of ovules to carpels when ectopically expressed in flowers, as previously reported for the related gene KNAT2. In contrast to KNAT2, loss of which confers no phenotype, we show using inducible RNA interference and mutational analysis that progressive loss of STM causes floral phenotypes ranging from reduced formation of placental tissues and inhibited carpel fusion to complete loss of carpel development. These effects result neither from failure to establish the central stem cell niche nor from reduced floral homeotic gene expression, but rather indicate a specific requirement for STM in carpel initiation, as further supported by the loss of leafy carpelloid features in stm leafy double mutants. Activation of carpel development by STM is independent of LEAFY and WUSCHEL, but requires the function of AGAMOUS. The essential role for STM in carpel development, together with its previous reported role in the SAM, shows that, despite the existence of several partially redundant paralogous genes, STM provides the critical KNOX function in the development of both vegetative and reproductive meristematic tissues. PMID- 17461794 TI - Self-incompatibility of Prunus tenella and evidence that reproductively isolated species of Prunus have different SFB alleles coupled with an identical S-RNase allele. AB - Many species of Prunus display an S-RNase-based gametophytic self-incompatibility (SI), controlled by a single highly polymorphic multigene complex termed the S locus. This comprises tightly linked stylar- and pollen-expressed genes that determine the specificity of the SI response. We investigated SI of Prunus tenella, a wild species found in small, isolated populations on the Balkan peninsula, initially by pollination experiments and identifying stylar-expressed RNase alleles. Nine P. tenella S-RNase alleles (S(1)-S(9)) were cloned; their sequence analysis showed a very high ratio of non-synonymous to synonymous nucleotide substitutions (K(a)/K(s)) and revealed that S-RNase alleles of P. tenella, unlike those of Prunus dulcis, show positive selection in all regions except the conserved regions and that between C2 and RHV. Remarkably, S(8)-RNase, was found to be identical to S(1)-RNase from Prunus avium, a species that does not interbreed with P. tenella and, except for just one amino acid, to S(11) of P. dulcis. However, the corresponding introns and S-RNase-SFB intergenic regions showed considerable differences. Moreover, protein sequences of the pollen expressed SFB alleles were not identical, harbouring 12 amino-acid replacements between those of P. tenella SFB(8) and P. avium SFB(1). Implications of this finding for hypotheses about the evolution of new S-specificities are discussed. PMID- 17461795 TI - Cholesterol-sensitive Cdc42 activation regulates actin polymerization for endocytosis via the GEEC pathway. AB - Glycosyl-phosphatidylinositol (GPI)-anchored proteins (GPI-APs) are present at the surface of living cells in cholesterol dependent nanoscale clusters. These clusters appear to act as sorting signals for the selective endocytosis of GPI APs via a Cdc42-regulated, dynamin and clathrin-independent pinocytic pathway called the GPI-AP-enriched early endosomal compartments (GEECs) pathway. Here we show that endocytosis via the GEECs pathway is inhibited by mild depletion of cholesterol, perturbation of actin polymerization or overexpression of the Cdc42/Rac-interactive-binding (CRIB) motif of neural Wiskott-Aldrich syndrome protein (N-WASP). Consistent with the involvement of Cdc42-based actin nanomachinery, nascent endocytic vesicles containing cargo for the GEEC pathway co-localize with fluorescent protein-tagged isoforms of Cdc42, CRIB domain, N WASP and actin; high-resolution electron microscopy on plasma membrane sheets reveals Cdc42-labelled regions rich in green fluorescent protein-GPI. Using total internal reflection fluorescence microscopy at the single-molecule scale, we find that mild cholesterol depletion alters the dynamics of actin polymerization at the cell surface by inhibiting Cdc42 activation and consequently its stabilization at the cell surface. These results suggest that endocytosis into GEECs occurs through a cholesterol-sensitive, Cdc42-based recruitment of the actin polymerization machinery. PMID- 17461796 TI - The cellular trafficking of the secretory proprotein convertase PCSK9 and its dependence on the LDLR. AB - Mutations in the proprotein convertase PCSK9 gene are associated with autosomal dominant familial hyper- or hypocholesterolemia. These phenotypes are caused by a gain or loss of function of proprotein convertase subtilisin kexin 9 (PCSK9) to elicit the degradation of the low-density lipoprotein receptor (LDLR) protein. Herein, we asked whether the subcellular localization of wild-type PCSK9 or mutants of PCSK9 and the LDLR would provide insight into the mechanism of PCSK9 dependent LDLR degradation. We show that the LDLR is the dominant partner in regulating the cellular trafficking of PCSK9. In cells lacking the LDLR, PCSK9 localized in the endoplasmic reticulum (ER). In cells expressing the LDLR, PCSK9 sorted to post-ER compartments (i.e. endosomes in cell lines and Golgi apparatus in primary hepatocytes), where it colocalized with the LDLR. In cell lines, PCSK9 also colocalized with the LDLR at the cell surface, requiring the presence of the C-terminal Cys/His-rich domain of PCSK9. We provide evidence that PCSK9 promotes the degradation of the LDLR by an endocytic mechanism, as small interfering RNA mediated knockdown of the clathrin heavy chain reduced the functional activity of PCSK9. We also compared the subcellular localization of natural mutants of PCSK9 with that of the wild-type enzyme in human hepatic (HuH7) cells. Whereas the mutants associated with hypercholesterolemia (S127R, F216L and R218S) localized to endosomes/lysosomes, those associated with hypocholesterolemia did not reach this compartment. We conclude that the sorting of PCSK9 to the cell surface and endosomes is required for PCSK9 to fully promote LDLR degradation and that retention in the ER prevents this activity. Mutations that affect this transport can lead to hyper- or hypocholesterolemia. PMID- 17461797 TI - Nuclear functions of the Arf guanine nucleotide exchange factor BRAG2. AB - BRAG2 is a guanine nucleotide exchange factor for the GTPase Arf6 that cycles between the cytoplasm and nucleus in a CRM1/exportin1-dependent manner. Despite its presence in the nucleus, nuclear functions have not previously been described. Here, we show that depletion of endogenous BRAG2 by RNAi leads to an increased number of Cajal bodies (CBs), and altered structure of nucleoli, as indicated by less focal fibrillarin staining. This result was surprising given that nuclear BRAG2 is diffusely distributed throughout the nucleoplasm and is not concentrated within nucleoli at steady state. However, we found that ectopic expression of the nuclear GTPase PIKE/AGAP2 causes both BRAG2 and the CB marker coilin to accumulate in nucleoli. Neither the GTPase activity of PIKE nor the nucleotide exchange activity of BRAG2 is required for this nucleolar concentration. Increased levels of exogenous BRAG2 in nucleoli result in a redistribution of fibrillarin to the nucleolar periphery, supporting a role for BRAG2 in regulating nucleolar architecture. These observations suggest that, in addition to its role in endocytic regulation at the plasma membrane, BRAG2 also functions within the nucleus. PMID- 17461798 TI - The peroxin PEX14 of Neurospora crassa is essential for the biogenesis of both glyoxysomes and Woronin bodies. AB - In the filamentous fungus Neurospora crassa, glyoxysomes and Woronin bodies coexist in the same cell. Because several glyoxysomal matrix proteins and also HEX1, the dominant protein of Woronin bodies, possess typical peroxisomal targeting signals, the question arises as to how protein targeting to these distinct yet related types of microbodies is achieved. Here we analyzed the function of the Neurospora ortholog of PEX14, an essential component of the peroxisomal import machinery. PEX14 interacted with both targeting signal receptors and was localized to glyoxysomes but was virtually absent from Woronin bodies. Nonetheless, a pex14Delta mutant not only failed to grow on fatty acids because of a defect in glyoxysomal beta-oxidation but also suffered from cytoplasmic bleeding, indicative of a defect in Woronin body-dependent septal pore plugging. Inspection of pex14Delta mutant hyphae by fluorescence and electron microscopy indeed revealed the absence of Woronin bodies. When these cells were subjected to subcellular fractionation, HEX1 was completely mislocalized to the cytosol. Expression of GFP-HEX1 in wild-type mycelia caused the staining of Woronin bodies and also of glyoxysomes in a targeting signal dependent manner. Our data support the view that Woronin bodies emerge from glyoxysomes through import of HEX1 and subsequent fission. PMID- 17461799 TI - Nup53p is a target of two mitotic kinases, Cdk1p and Hrr25p. AB - Nuclear pore complexes (NPCs) form channels across the nuclear envelope and provide the sole sites of molecular exchange between the cytoplasm and nucleoplasm. The NPC is a target of a number of post-translational modifications, including phosphorylation, yet the functions of these modifications are ill defined. Here, we have investigated the mitotic specific phosphorylation of a yeast nucleoporin Nup53p. Two kinases were identified that phosphorylate Nup53p: the mitotic kinase Cdk1p/Cdc2p/Cdc28p and the casein kinase Hrr25p. Hrr25p was identified by screening 119 yeast kinases for their ability to phosphorylate Nup53p in vitro. Conditional alleles of Hrr25p support the conclusion that Hrr25p phosphorylates Nup53p in vivo. We further demonstrated using solution binding and affinity purification assays, that Hrr25p directly binds Nup53p in an interaction that is destabilized by the phosphorylation of Nup53p. Consistent with this observation, we observed that Hrr25p moves between distinct locations in the cell during the cell cycle including the nucleus, the cortex of the emerging bud and the spindle pole bodies. Cdk1p also contributes to Nup53p phosphorylation as specific inhibition of Cdk1p or mutation of Cdk1p consensus sites partially blocked its phosphorylation. The ability of nup53 alleles containing Cdk1p site mutations to complement synthetic defects of nup53 Delta nup170 Delta strains is linked to a function for Nup53p in the spindle assembly checkpoint. PMID- 17461800 TI - Intracellular trafficking in the trypanosomatids. AB - Trypanosomes are members of the kinetoplastida, a group of divergent protozoan parasites responsible for considerable morbidity and mortality worldwide. These organisms have highly complex life cycles requiring modification of their cell surface together with engagement of immune evasion systems to effect survival; both processes intimately involve the membrane trafficking system. The completion of three trypanosomatid and several additional protist genomes in the last few years is providing an exciting opportunity to evaluate, at the molecular level, the evolution and diversity of membrane trafficking across deep evolutionary time as well as to analyse in unprecedented detail the membrane trafficking systems of trypanosomes. PMID- 17461801 TI - Pigmented hypertrichotic dermatosis and insulin dependent diabetes: manifestations of a unique genetic disorder? AB - A novel pigmented dermatosis was observed in four unrelated boys, three of whom had insulin-dependent diabetes. Three patients were the offspring of consanguineous parents. All four boys had pigmented hypertrichotic patches or induration on the upper inner thighs, with variable involvement of the genitalia, trunk, and limbs. Two boys had episcleritis and orbital proptosis with similar facies and musculoskeletal abnormalities including clinodactyly, flat feet, and short stature. One child had paraaortic and inguinal lymphadenopathy and three patients had an enlarged liver and spleen. A large, swollen pancreas was observed on ultrasound imaging in one patient with insulin dependent diabetes who also had echocardiographic evidence of pericardial inflammation. Three boys had elevated laboratory markers of inflammation. Biopsy specimens from the skin and orbit showed a chronic inflammatory cell infiltrate composed of polyclonal lymphocytes, histiocytes, and plasma cells; fibrosis was observed in two patients, one of whom had previously received radiation therapy to the orbit. Two boys responded to treatment with subcutaneous interferon-alpha, combined with a short course of oral prednisone in the child without diabetes. We believe these inflammatory pigmented skin lesions represent a unique dermatosis associated with diabetes mellitus and systemic disease. The pathogenesis is unknown. The presence of consanguinity in three of four families, and similar dysmorphic features in two boys, suggest a genetic disorder, possibly with autosomal recessive inheritance. PMID- 17461802 TI - Skin cancer prevention in the primary care setting: assessment using a standardized patient. AB - Our objective was to utilize the standardized patient technique in assessing the ability of primary care physicians to identify and counsel primary prevention for patients at high risk for skin cancer. A secondary goal was to test the feasibility of this technique as a measure of actual physician behaviors in the outpatient setting. We used a convenience sample of 15 primary care physicians. The standardized patient was an 18-year-old woman with skin phototype I. She presented to physicians as needing a general physical examination for a summer lifeguard job at a beach. She stated a family history of skin cancer. Physician performances were rated using a standard checklist completed by the standardized patient following each visit. We found that none of the physicians asked questions specifically related to skin phototype or sun exposure habits such as childhood sunburns. Only 13% asked about mole changes. For counseling, 67% of physicians recommended sunscreen use; only 7% discussed sunscreen types or procedures for effective use. Only 13% counseled other skin protective behaviors. No significant differences by physician gender were found in these areas; however, female physicians counseled more global health behaviors than male physicians (p < or = 0.01). Our pilot data suggest that little skin cancer primary prevention counseling is performed for high-risk patients. The standardized patient technique worked well in obtaining outcome data for physicians' preventive practices. PMID- 17461803 TI - Health provider factors associated with nonattendance in pediatric dermatology ambulatory patients. AB - Nonattendance for dermatology appointments disrupts the management of medical delivery and leads to inefficient allocation of resources and lost revenue. The factors that determine nonattendance in pediatric dermatology patients have not been well documented. We investigated health provider factors for nonattendance in pediatric dermatology patients. We assessed the effects of waiting time for an appointment and the timing of the appointment (during the day, week, and year) on nonattendance proportions during a 1 year period. Chi-squared tests were used to analyze statistically significant differences of categorical variables. Logistic regression was used for multivariate analyses. A total of 1696 children visits were included in the study. The overall rate of nonattendance at the dermatology clinic was 30.5%. Nonattendance was 29.7% during the periods between 8 a.m. to 1 p.m. and 3 p.m. to 7 p.m. and 40.7% during the periods between 1 p.m. to 3 p.m. and 7 p.m. to 8 p.m. (p = 0.013). Nonattendance was 21.1% when the waiting time for an appointment was short (1-7 days), 32.5% when it was intermediate (8-14 days) and 43.5% when the wait time was long (15 days and above) (p-value < 0.001). A multivariate logistic regression model demonstrated that the hour of the day and the waiting time for an appointment were significantly associated with nonattendance (p value = 0.009, p value < 0.001, respectively). We conclude that in children attending a dermatology clinic, health provider factors that determine nonattendance include the waiting time for an appointment and the hour of the appointment within the day. PMID- 17461804 TI - The use of alternative medicine in pediatric patients with atopic dermatitis. AB - Alternative medicine has been defined as forms of therapy or examination that have no scientific basis and for which no effective or diagnostic reliability has been demonstrated by scientific methods. The use of complementary or alternative medicine is increasing and controlled clinical trials on the subject are few. We performed a questionnaire-based study of 80 pediatric patients with atopic dermatitis. This questionnaire assessed the duration of treatment, the reason(s) for trying alternative therapy, the approximate cost and the success of the treatment, the duration of the childhood eczema, and whether the child had ever required hospital admission for eczema. Of the total, 34 (42.5%) patients had used alternative medicine. Herbal remedies and homeopathy were used most often. Most treatments were reported to show no benefit and in three instances deterioration was reported. This study has prompted us to enquire routinely regarding alternative medicine use. Alternative therapies are subject to minimal regulation and have been associated with serious side effects. We would recommend enquiries regarding alternative medicine use in all pediatric dermatology patients. PMID- 17461805 TI - Hemorrhagic subcutaneous nodules: an initial clinical sign of hemophilia A. AB - A 9-month-old boy presented with multiple hemorrhagic subcutaneous nodules after treatment with oral antibiotics for an upper respiratory tract infection. Excessive bleeding after an incisional biopsy led to a diagnosis of hemophilia A. While this was an unexpected presentation of hemophilia, screening laboratory coagulation studies prior to biopsy would have revealed the etiology. Similar presentations should alert dermatologists to suspect an underlying coagulation defect prior to biopsy, thus averting bleeding complications. PMID- 17461806 TI - Linear cutaneous lupus erythematosus in the lines of Blaschko. AB - Eleven patients with linear cutaneous lupus erythematosus following the lines of Blaschko have been previously reported in the literature. We describe a child with this entity following Blaschko lines on the trunk. The patient responded to oral hydroxychloroquine therapy combined with topical flurandrenolide tape resulting in resolution with atrophic scarring. The clinical and histological features of linear cutaneous lupus erythematosus are discussed and a review of the literature is presented. PMID- 17461807 TI - Epstein-Barr virus-associated genital ulcers: an under-recognized disorder. AB - Infectious mononucleosis is the best-known syndrome associated with primary Epstein-Barr virus infection. Although a variety of cutaneous and mucosal manifestations are recognized in infectious mononucleosis, genital ulcers have only rarely been described. We describe an otherwise healthy 14-year-old girl in whom painful genital ulcers developed during an episode of serologically confirmed primary Epstein-Barr virus infection. Clinical, serologic, and histopathologic evaluation failed to disclose evidence of any other etiologic explanation for her lesions. The patient remains well, without recurrence. To date, only 13 instances of genital ulceration in females attributable to Epstein Barr virus infection have been reported. PMID- 17461808 TI - Eruptive post-chemotherapy in situ melanomas and dysplastic nevi. AB - A 22-year-old white man without a personal or family history of atypical nevi had received chemotherapy for pre-B-cell acute lymphocytic leukemia at age 17 that included L-asparaginase, prednisone, methotrexate, mercaptopurine, daunorubicin, and cytoxan. Two to three months after completing maintenance chemotherapy, the patient reports he developed many moles, which remained stable for approximately 2 years. Upon examination, two dark, atypical appearing plaques with irregular borders and numerous pink papules of varying shapes and sizes were noted on his chest, back, and abdomen. Histology of specimens of both types of lesions revealed three moderately atypical compound dysplastic melanocytic nevi and three in situ melanomas. The lesions with only features of dysplastic nevi exhibited dermal fibrosis, cytologic atypia, junctional shoulders, lentiginous spread, and focal pigmentation. The lesions with in situ melanomas in addition demonstrated pagetoid spread, extension down adnexal structures, and more severe cytologic atypia. Malignant melanoma has been associated with chronic immunosuppression, and benign nevi have been reported to erupt after chemotherapy. We report an occurrence of multiple eruptive dysplastic nevi and in situ melanomas appearing shortly after completion of chemotherapy. PMID- 17461809 TI - The hair collar sign as a marker for neural tube defects. AB - The hair collar sign has been described as a marker of cranial dysraphism, including encephaloceles, meningoceles, and heterotropic brain tissue. This report describes a male born with a hair collar sign who subsequently was found to have a Klippel-Feil anomaly, diastematomyelia, multiple segmental spine and rib anomalies, and a translocation between chromosomes 1 and 4; t(1;4) (q44;q10). While not previously documented, spine abnormalities may be an associated abnormality with the hair collar sign. Physicians should consider radiographic evaluation of the spine in all newborns with a hair collar sign, as well as genetic screening. PMID- 17461810 TI - Cerebriform intradermal nevus. AB - Cerebriform intradermal nevus is a rare form of cutis verticis gyrata. Clinically it manifests as a scalp deformity resembling the surface of the brain, with cerebriform morphologic characteristics. Degeneration into malignant melanoma has been reported. Herein, a cerebriform intradermal nevus of the scalp in a 7-year old girl is reported. The clinical and histopathologic presentations of cerebriform intradermal nevus are described and the therapeutic possibilities discussed. PMID- 17461811 TI - Mosaic chromosome 6 trisomy in an epidermal nevus. AB - We report a 2-year-old boy with a linear epidermal nevus. Mosaicism for chromosome 6 in skin fibroblasts of affected skin was discovered. Trisomy 6 has not been previously implicated as an isolated finding in epidermal nevi or cutaneous mosaicism. PMID- 17461812 TI - Linear morphea presenting as acquired unilateral edema. AB - We describe a 2-year-old African-American boy with a 4-month history of gradually worsening unilateral edema that was initially noted on his left hand and then approximately 2 weeks later on his left lower extremity. In addition, linear hypopigmented patches were noted along the left forearm and leg, with no appreciable scarring or induration. The edema on the left-hand side of his body progressed so that he developed tense bullae on his left hand. Two months later, the hypopigmented patches were indurated and bound-down, especially over the left groin and thigh. A biopsy specimen from this area showed features characteristic of morphea. In this patient, dilated lymphatic channels secondary to the sclerosis of the morphea caused the bullae. Bullous morphea is a rare condition. We were unable to find any reports its occurrence in children under 18 with associated lymphedema. This entity should be included in the differential for acquired unilateral edema in children. PMID- 17461813 TI - Onychomycosis in a premature infant caused by Candida parapsilosis. AB - We report onychomycosis caused by Candida parapsilosis in a 35-day-old premature infant. To our knowledge, this represents the youngest instance of an isolated lesion of the nail plate without involvement of glabrous skin, caused by C. parapsilosis in this age group to be reported in the literature. PMID- 17461814 TI - Dermatomyositis with a pityriasis rubra pilaris-like eruption: an uncommon cutaneous manifestation in dermatomyositis. AB - A pityriasis rubra pilaris-like eruption has been described in patients with dermatomyositis. We describe an 11-year-old girl with dermatomyositis who had additional clinical findings of pityriasis rubra pilaris. Over a year, she developed muscle weakness, increasing fatigue, and a markedly elevated creatinine kinase level in addition to her cutaneous eruption and was seen in our clinic for these complaints. A year earlier, when a generalized, scaly erythematous eruption had appeared, she had been diagnosed as pityriasis rubra pilaris clinically and histopathologically. Dermatologic examination found scaling erythematous plaques involving the trunk and upper and lower extremities. Islands of unaffected skin were intermingled with erythematous plaques that were characteristic of pityriasis rubra pilaris. A skin biopsy specimen showed the findings of dermatomyositis and that diagnosis was made. The laboratory findings, electromyographic pattern, and muscle biopsy were also consistent with dermatomyositis. Her presentation is interesting, as she had been diagnosed as pityriasis rubra pilaris both clinically and histopathologically 1 year earlier and, although the cutaneous lesions had not changed, a diagnosis of dermatomyositis was made a year later. PMID- 17461815 TI - Lymphotropic adamantinoid trichoblastoma. AB - We report a lymphotropic adamantinoid trichoblastoma in a 14-year-old girl. This is a rarely reported skin adnexal tumor in adolescent patients. The clinical, histologic, and immunohistochemical features of this tumor are described. PMID- 17461816 TI - Widespread porokeratotic eccrine ostial and dermal duct nevus along Blaschko lines. AB - Porokeratotic eccrine ostial and dermal duct nevus is a congenital hamartoma of possible eccrine origin with no malignant potential. It is usually localized at the extremity of a single limb, while wider systematized distribution has rarely been documented. A child with an unusually widespread nevus following Blaschko lines is reported. The disorder had a striking presentation in the form of a systematized linear epidermal nevus composed of multiple tiny filiform keratotic spines, which histologically corresponded to columns of porokeratosis. Serial histopathologic sections of a 4-mm punch biopsy specimen barely demonstrated an anatomic relationship between the porokeratotic columns and the underlying acrosyringeal duct. This report provides further evidence of porokeratotic eccrine ostial and dermal duct nevus being distributed along Blaschko lines, thus confirming it is a peculiar epidermal nevus due to a mosaic cutaneous condition. PMID- 17461817 TI - Transilluminator burns in the neonatal intensive care unit: a mimicker of more serious disease. AB - We report an outbreak of erythematous to purpuric papules and vesicles in four neonates whose lesions arose within a 72-hour period in a neonatal intensive care unit. As isolated lesions, the clinical presentation mimicked cutaneous aspergillosis. Ultimately, because of the number of patients with similar lesions, an exogenous source was suspected and determined to be a faulty transillumination device. PMID- 17461818 TI - Generalized erythrodermic pemphigus foliaceus in a child and its successful response to rituximab treatment. AB - Pemphigus foliaceus is an autoimmune disease that clinically manifests with cutaneous blisters of the superficial skin. The nonendemic or sporadic form of this entity is rare in children and typically presents with a milder, more localized rash that usually follows a benign course of short duration. We describe an affected patient atypical in both her young age and the severity of skin findings. Our patient presented with a full body exfoliative erythroderma at 21 months of age. After an extensive work-up to determine the etiology of her exfoliative erythroderma, direct and indirect immunofluorescence studies confirmed the diagnosis of pemphigus foliaceus. Rituximab therapy was initiated based on the patient's refractory disease course to multiple immunosuppressive agents. Rituximab is a therapeutic monoclonal antibody targeting CD20, an integral membrane protein highly expressed on the surface of pre-B lymphocytes and activated mature B lymphocytes. The patient's skin exhibited marked clinical improvement after the start of rituximab infusions over 12 weeks. Her initial desmoglein 1 antibody level was greater than 1:1280, which decreased to 1:16 after seven rituximab treatments. She has had no skin flares since initiating treatment with rituximab therapy. Based on this clinical and serologic response, the use of rituximab may be helpful in the treatment of pediatric pemphigus foliaceus refractory to mainstays of therapy. PMID- 17461819 TI - Pulsed dye laser therapy for pediatric warts. AB - Viral wart infections constitute one of the most common pediatric skin diseases, and various modalities have been used to manage them. Although pulsed dye laser therapy is known to be a safe and efficacious modality, the reported cure rates for this method have varied, and no studies have reported treatment of pediatric patients alone. This prospective, nonblinded, nonrandomized study was performed to evaluate the efficacy and safety of pulsed dye laser therapy for pediatric warts. We found that this method is safe, relatively effective, and worth considering as an additional therapeutic option for viral warts in children, although not as a first-line therapy. PMID- 17461820 TI - Congenital generalized hypomelanosis and immunodeficiency in a black child. PMID- 17461821 TI - What syndrome is this? Apert syndrome. PMID- 17461822 TI - What syndrome is this? Ehlers-Danlos syndrome type VIII. PMID- 17461823 TI - Modified curettage technique for molluscum contagiosum. AB - Molluscum contagiosum is a frequently occurring, virally induced cutaneous condition that affects infants, children, and adults. Although historically considered a self-limiting entity, many patients experience prolonged infections, often resistant to varied therapeutic interventions. Topical treatments achieve complete clearance in only a limited number of patients. Because of the associated pain, affected children are characteristically uncompliant to commonly used destructive modalities. We describe a modification of the traditional curettage technique to remove lesions of molluscum contagiosum, which has proven to be effective and well tolerated by a significant number of patients in our practice. PMID- 17461824 TI - Collagenase application for amputation in a preterm. PMID- 17461825 TI - Localized retroauricular morphea in a boy with Lennox-Gastaut syndrome and hypospadias. PMID- 17461826 TI - Acquired aquagenic keratoderma. PMID- 17461827 TI - Case detection rates of molluscum contagiosum in childhood. PMID- 17461828 TI - Extensive pityriasis alba in a nonatopic child. PMID- 17461829 TI - Solitary morphea profunda with a prominent mucinous deposit. PMID- 17461831 TI - Abstracts of the 21st European Immunogenetics and Histocompatibility Conference. Barcelona, Spain. May 5-8, 2007. PMID- 17461833 TI - Comparative SEM study of the marginal adaptation of white and grey MTA and Portland cement. AB - The use of a suitable substance that prevents egress of potential contaminants into the periapical tissues is important in endodontic surgery. The aim of the present study was to compare the marginal adaptation of three root-end filling materials (white mineral trioxide aggregate (MTA), grey MTA and Portland cement), using scanning electron microscopy. Seventy-five single-rooted extracted human teeth were used. The canals were instrumented and filled with gutta-percha. Following root-end resection and cavity preparation, root-end cavities were filled with white MTA, grey MTA or Portland cement. Using a diamond saw, roots were longitudinally sectioned into two halves. Under scanning electron microscopy, the gaps between the material and dentinal wall were measured. The data were analysed using Kruskal-Wallis test. The mean of the gap in grey MTA, white MTA and Portland cement was 211.6, 349 and 326.3 microm, respectively. The results indicate that the gap between grey MTA and the dentinal wall is less than other materials, but there was no significant difference between the materials tested in this study (P > 0.05). PMID- 17461834 TI - In vitro evaluation of the accuracy of five different electronic apex locators for determining the working length of endodontically retreated teeth. AB - The aim of this study was to evaluate the accuracy of five electronic apex locators (EALs) in determining the working length (WL) of teeth after removal of the root canal obturation materials. A total of 32 extracted straight, single rooted teeth were used. The actual canal length (AL) was determined and the WL was established by subtracting 0.5 mm from the AL. The root canals were instrumented and divided into two groups. One group (n = 6) served as control, while the other group (n = 26) was the experimental group. In the experimental group, the root canals were obturated using vertically compacted gutta-percha with AH 26 sealer. In both groups, the access cavities were restored with a provisional restoration and stored for 15 days at 37 degrees C and 100% humidity. The root canal obturation material was removed, and the teeth were then mounted in an experimental apparatus. Five EALs were used: Dentaport ZX, ProPex, Foramatron D10, Apex NRG and Apit 7. For the electronic measurement of canal length, a size 25 K-file was used. During measurement, the canal was irrigated with 2.5% sodium hypochlorite. The difference (D) between the AL and the electronically determined length (EDL), AL-EDL, was calculated and recorded for each measurement. Data were analysed by two-way anova and Fisher's PLSD test. In both groups, statistically significant differences were found among the EALs (P < 0.01). In conclusion, the Dentaport ZX, ProPex and Foramatron D10 were more accurate than the other two EALs in determining the WL in teeth after removal of the root canal obturation materials. However, the Apex NRG and Apit 7 were also reliable for determination of the WL in the majority of the cases. PMID- 17461835 TI - Canal configuration in the mesio-buccal root of maxillary first molar teeth of a Jordanian population. AB - The purpose of this study was to investigate the canal configuration in the mesio buccal root of maxillary first molar teeth of a Jordanian population using a clearing technique. One hundred extracted maxillary first molars were collected from Jordanian patients. After a standard access preparation, canals were stained, decalcified, dehydrated, and then rendered clear. The prevalence of a second canal in the mesio-buccal root was 77.32%. Types IV and II canal systems were the most common types with prevalence of 35.05% and 27.83%, respectively. Additionally, 28.86% showed lateral canals mostly located in the apical third and 37.11% had intercanal communications, mainly in the middle third of the root. It was concluded that the prevalence of mesio-palatal canals was high and Types IV and II canal systems were the most prevalent. PMID- 17461836 TI - Leakage evaluation of three different root canal obturation techniques using electrochemical evaluation and dye penetration evaluation methods. AB - The aim of this study was to compare the apical sealing ability of three different obturation techniques using an electrochemical evaluation and evaluating dye penetration. One hundred and thirty-two maxillary anterior teeth were randomly divided into six groups. There were 20 teeth in each group. The teeth were obturated as follows: Groups 1 and 4 with Thermafil, Groups 2 and 5 with System B, and Groups 3 and 6 with cold lateral condensation (CLC). The apical leakage in these groups was evaluated using an electrochemical method for Groups 1, 2, 3 and a dye penetration method for Groups 4, 5 and 6. In both methods, the lowest mean leakage values were observed for Thermafil and the highest were observed for the CLC groups. The difference between Thermafil and CLC was statistically significant (P < 0.05). In both groups, System B fillings showed moderate leakage and the difference was not significant with Thermafil and CLC groups. PMID- 17461837 TI - Evaluation of the thermoplasticity of different gutta-percha cones and Resilon. AB - The goal of this study was to evaluate the thermoplasticity of conventional and thermoplastic gutta-percha and Resilon, a polyester polymer-based material. Specimens with standardised dimensions were made from the following materials: conventional and thermoplastic gutta-percha (Dentsply), conventional and thermoplastic gutta-percha (Endopoints) and Resilon. After 24 h, the specimens were placed in water at 70 degrees C for 60 s, and thereafter positioned between two glass slabs. Each set was compressed by a 5-kg weight. Digital images of the specimens before and after compression were obtained and analysed. The thermoplasticity of each material was confirmed by the difference between final and initial areas of each sample. The data were analysed statistically by anova and Tukey's test at a 5% significance level. Resilon had the highest thermoplasticity means (P < 0.05). Among the gutta-percha cones, Endopoints TP (thermoplastic) presented the highest thermoplasticity means and differed significantly from the other commercial brands (P < 0.05). Resilon had good thermoplasticity, endorsing its use as a thermoplastic root canal filling material. PMID- 17461838 TI - Endodontic treatment of mandibular incisors with two root canals: report of two cases. AB - The root canal anatomy of mandibular incisors can present a number of variations, including multiple canals. Two case reports are presented to illustrate the serendipitous discovery and successful non-surgical endodontic management of complex canal systems in mandibular incisors. In both cases, all four mandibular incisors had two canals each. PMID- 17461839 TI - Regression of a lesion resembling pyogenic granuloma after endodontic treatment. AB - A rare case of regression of a lesion resembling pyogenic granuloma after endodontic treatment of a maxillary lateral incisor in a 16-year-old female patient is reported. After clinical and radiographic examination, the maxillary right lateral incisor was endodontically treated in two visits. At the second visit, clinical regression of the exophytic lesion was evident and the size of this lesion had been greatly decreased, so was not biopsied. After 3 weeks, the exophytic lesion had totally disappeared. This case report indicates that surgical excision of exophytic lesions resembling pyogenic granuloma is not necessary in all situations. PMID- 17461840 TI - Repair of large periapical radiolucent lesions of endodontic origin without surgical treatment. AB - The aim of this paper is to present two case reports of pulp necrosis and radiolucent periapical lesions, which were treated without surgical treatment. The first was a mandibular molar with periapical lesion of endodontic origin extending towards the furcation in a 20-year-old woman, and the second affected a maxillary right lateral incisor with a large periapical lesion in a 22-year-old woman. The endodontic treatments were carried out in two sessions, with crown down instrumentation, irrigation with 2.5% sodium hypochlorite and intracanal medication with calcium hydroxide paste. After 30 days, the root canals were filled with gutta-percha and Sealapex sealer by the lateral condensation technique. The clinical and radiographic examination after 1 year revealed complete repair. The appropriate diagnosis of lesions of endodontic origin and the treatment and obturation of the infected canals allowed complete repair of these large radiolucent periapical lesions without surgical treatment. PMID- 17461841 TI - Treatment of a second maxillary molar with six canals. AB - Variations in the dental anatomy are found in all teeth. Knowledge of these variations, particularly concerning the location and treatment of all canals, is very important for the success of the endodontic therapy. The purpose of this study is to present a clinical case of a maxillary second molar with three palatal canals, two mesio-buccal and one disto-buccal canal. This report serves to remind clinicians that such anatomical variations should be taken into account during endodontic treatment of the maxillary molars. PMID- 17461847 TI - Multivitamin supplementation in HIV-positive pregnant women: impact on depression and quality of life in a resource-poor setting. AB - OBJECTIVES: The primary objective of this study was to examine the effect of vitamin supplementation on health-related quality of life and the risk of elevated depressive symptoms comparable to major depressive disorder (MDD) in HIV positive pregnant women in Dar es Salaam, Tanzania. METHODS: From April 1995 to July 1997, 1078 HIV-positive pregnant women were enrolled in a randomized controlled trial. We examined the effects of vitamin supplementation on quality of life and the risk of elevated depressive symptoms, assessed longitudinally every 6-12 months. RESULTS: A substantial prevalence of elevated depressive symptoms (42%) was observed in HIV-positive pregnant women. Multivitamin supplementation (B-complex, C and E) demonstrated a protective effect on depression [relative risk (RR)=0.78; P=0.005] and quality of life [RR=0.72 for social functioning (P=0.001) and vitality (P=0.0001); RR=0.70 for role-physical (P=0.002)]; however, vitamin A showed no effect on these outcomes. CONCLUSIONS: Multivitamin supplementation (B-complex, C and E) resulted in a reduction in risk of elevated depressive symptoms comparable to MDD and improvement in quality of life in HIV-positive pregnant women in Tanzania. PMID- 17461848 TI - Improved virological response to highly active antiretroviral therapy in HIV-1 infected patients carrying the CCR5 Delta32 deletion. AB - BACKGROUND: Patients heterozygous for the C-C chemokine receptor 5 (CCR5) Delta32 deletion spontaneously progress less rapidly to AIDS and death than do wild-type patients. We investigated whether the CCR5 Delta32 deletion has an impact on immunological, virological and clinical responses to highly active antiretroviral therapy (HAART) in HIV-1-infected patients. PATIENTS AND METHODS: We included in the study 565 HIV-1-infected patients from the French HIV-1 infected cohort with documented date of seroconversion (SEROCO)/haemophiliacs HIV-1 infected (HEMOCO) cohorts, who started HAART after 1996. We investigated virological responses to HAART at 6 months (defined as a plasma HIV-1 RNA measurement below the threshold of detection or a 2 log HIV-1 RNA decrease) and at 12 months (defined as a plasma HIV-1 RNA measurement below the threshold of detection) and clinical response to HAART by Kaplan-Meier survival curves, with AIDS and death as outcomes. RESULTS: The Delta32 heterozygous patients (n=83; 15%) had a better virological response to HAART than wild-type patients (73 vs 53% at 6 months, P=0.01; and 60 vs 44% at 12 months, P=0.01). This better virological response was still observed after adjustment for antiretroviral status (whether or not patients were naive to antiretroviral therapy), year of HAART initiation, number of new antiretroviral drugs and baseline viral load. There was no statistical difference between heterozygous patients and wild-type patients in terms of survival and AIDS-free survival. CONCLUSIONS: CCR5 Delta32 heterozygous patients were more likely to have a virological response to HAART than wild-type patients at 6 and 12 months. However, this virological response did not produce better immunological and clinical responses. The long-term impact of the Delta32 deletion on survival in HIV-1-infected treated patients should be investigated in a meta-analysis. PMID- 17461849 TI - Concomitant use of gastric acid-reducing agents is frequent among HIV-1-infected patients receiving protease inhibitor-based highly active antiretroviral therapy. AB - OBJECTIVE: The aim of the study was to assess the frequency of the concurrent use of gastric acid-reducing agents among HIV-1-infected patients treated with highly active antiretroviral therapy (HAART) combinations. METHODS: An anonymous, semistructured, self-administered questionnaire was consecutively distributed among HIV-1-infected patients at routine visits to specialized HIV clinics. The questionnaire contained 17 items asking specifically for information on current antiretroviral treatments and the use of gastric acid-reducing agents as well as demographic data. RESULTS: A total of 424 patients in 12 centres participated in the study: 85% were male, 88% were of German nationality, 82% were >35 years of age and 201 (47.4%) were receiving a protease inhibitor (PI)-containing HAART regimen. Of these, 74 (37%) had received an acid-reducing drug within the previous 6 months and 43 (58%) were currently still on it. Two-thirds of patients (64.9%) were treated with proton-pump inhibitors (pantoprazole, omeprazole or esomeprazole) and 56% of patients on PI-containing regimens had been taking these drugs for longer than 2 months and up to a maximum of 3 years. The majority of patients (77%) had received the prescription for the acid-reducing drugs from their HIV specialist and the remaining patients had received over the counter (OTC) medication or prescriptions from other medical personnel. CONCLUSIONS: A substantial subset of patients treated with HAART combinations, including those on PI-containing regimens, were using concomitant acid-reducing drugs, most often proton-pump inhibitors. As negative drug-drug interactions between some of the (boosted) PIs and gastric acid-reducing agents have recently been reported, HIV physicians should take this into account when prescribing PI-containing HAART combinations in order to avoid an additional risk of treatment failure. PMID- 17461850 TI - Inhibitory quotient as a prognostic factor of response to a salvage antiretroviral therapy containing ritonavir-boosted saquinavir. The CIVSA Study. AB - BACKGROUND: The addition of a low dose of ritonavir to protease inhibitors (PIs) has become a widespread strategy to improve PI pharmacokinetics. As resistance is a major barrier to long-term suppression, in salvage therapy genotype and/or phenotype scoring is currently used to predict the response. We evaluated the relationship between the saquinavir (SQV) inhibitory quotient (IQ) (virtual and genotypic) and virological response. METHODS: Eligible patients were on a PI containing highly active antiretroviral therapy (HAART) regimen excluding SQV and had a viral load >5000 HIV-1 RNA copies/mL. The PI was switched to SQV/ritonavir (RTV) 1000/100 mg twice a day (bid) and the same two backbone nucleoside reverse transcriptase inhibitors (NRTIs) were maintained at least until week 4, when the resistance test results became available. Genotype and virtual phenotype were determined at baseline, while the SQV trough plasma concentration was determined at week 4. RESULTS: Fifty-three patients were included in the study. Mean baseline viral load and CD4 count were 137,693 copies/mL and 263 cells/microL, respectively, the mean number of previous PIs was 2.3 and the mean number of protease gene mutations (PGMs) was 4.1. Using an on-treatment analysis, at week 16 the mean increase in CD4 count was 70.9 cells/microL, viral load was <200 copies/mL in 17 out of 37 patients (45.9%), and 30 out of 45 patients (66.7%) were considered virological responders (VRs) (viral load <200 copies/mL or viral load declined > or =1 log(10) at week 16). Median virtual phenotype was 1.3 (0.6 6.9). Baseline differences were detected between VR and non-VR populations: the mean numbers of PGMs were 3.2 and 5.8 (P<0.05), the mean numbers of SQV associated mutations were 2 and 3.8 (P<0.05), and the mean CD4 counts were 365.9 and 184.3 cells/microL (P<0.05), respectively. Mean SQV trough concentrations at week 4 were 1.1 and 1.0 microg/mL (not significant), and mean virtual IQs were 0.7 and 0.1 (P<0.01), respectively. Multivariate analysis showed that baseline PGMs >5 or SQV-associated mutations>5, virtual phenotype, baseline viral load >50,000 copies/mL, and virtual IQ <0.5, but not genotypic IQ, were the variables independently associated with non-VR. CONCLUSION: In heavily pretreated patients, the use of SQV virtual IQ or alternatively virtual phenotype, as well as PGMs, is a useful tool for the prediction of virological response. PMID- 17461851 TI - Prediction of clinical benefits of ritonavir-boosted TMC114 from treatment effects on CD4 counts and HIV RNA. AB - OBJECTIVE: The aim of the study was to predict reductions in progression to AIDS/death associated with the treatment benefit of antiretrovirals on CD4 counts and HIV RNA in the era of highly active antiretroviral therapy (HAART). DESIGN: The study design was a pooled analysis of two trials (POWER 1 and POWER 2) of optimized background treatment plus either TMC114/ritonavir (TMC114/r) or control protease inhibitor (CPI). METHODS: Across the two randomized trials (mean baseline CD4 count 114 cells/microL and HIV RNA 4.6 log(10) HIV-1 RNA copies/mL), CD4 counts rose by a mean of 98 cells/microL for TMC114/r 600/100 mg twice a day (bid) vs. 17 cells/microL for CPI at week 24; HIV RNA fell by a median of 1.90 and 0.49 log(10) copies/mL in the two groups, respectively. For the CD4 categorization method, cohort data on rates of progression to AIDS/death during HAART within preset CD4 ranges were used to predict rates of progression during TMC114/r and CPI treatment. For the regression method, data from clinical endpoint trials were used to correlate historical treatment effects on HIV RNA and CD4 with clinical benefits. RESULTS: The CD4 categorization method predicted a 48% reduction in clinical progression to AIDS/death for TMC114/r vs. CPI. The regression method predicted a 55% reduction [95% confidence interval (CI) 45-66%] in the hazard of progression to AIDS/death based on CD4 counts, with a 47% reduction (95% CI 38-53%) predicted from effects on HIV RNA. CONCLUSIONS: Independent methods generated similar predictions of a 47-55% reduction in progression to AIDS/death for TMC114/r vs. CPI treatment, based on the changes in CD4 counts and HIV RNA from the POWER 1 and POWER 2 trials. These methods could be used to estimate clinical benefits of other antiretrovirals. PMID- 17461852 TI - A double-blind, parallel-group, placebo-controlled, multicentre study of acetyl L carnitine in the symptomatic treatment of antiretroviral toxic neuropathy in patients with HIV-1 infection. AB - BACKGROUND: Nucleoside reverse transcriptase inhibitors (NRTIs) disrupt neuronal mitochondrial DNA synthesis, resulting in antiretroviral toxic neuropathy (ATN). Acetyl-L-carnitine (ALCAR) enhances neurotrophic support of sensory neurones, potentially providing symptom relief and nerve regeneration. OBJECTIVE: The objective of the study was to assess the safety and efficacy compared to placebo of intramuscular ALCAR in HIV-positive patients with symptomatic distal symmetrical polyneuropathy. METHODS: Ninety patients were enrolled and randomized to receive ALCAR [500 mg twice a day (bid); n=43] or placebo (n=47) intramuscularly twice daily for 14 days followed by 42 days of oral ALCAR 1000 mg bid. Assessment of pain was obtained using the Visual Analogue Scale (VAS), Total Symptom Score (TSS), Clinical Global Impression of Change, McGill Pain Questionnaire (MPQ), and the need for rescue analgesics. RESULTS: There was no statistically significant difference in changes in VAS over 14 days between groups for the intent-to-treat (ITT) population, but for the efficacy-evaluable (EE) population ALCAR treatment produced a significantly greater reduction in pain compared with placebo (P=0.022). The proportion of patients with an improvement in TSS over 14 days was greater in the ALCAR group compared with the placebo group, but the differences were not statistically significant. During the open-label phase, patients experienced an improvement in pain, as measured by the VAS, TSS and McGill Pain Questionnaire. CONCLUSION: ALCAR, administered twice a day intramuscularly to HIV-1-infected patients with symptomatic ATN, significantly reduced weekly mean pain ratings on the VAS compared with placebo. Treatment with oral ALCAR improved symptoms for the patient group as a whole. Intramuscular and oral ALCAR was generally safe and well tolerated. PMID- 17461853 TI - Incidence and causes of death in HIV-infected persons receiving highly active antiretroviral therapy compared with estimates for the general population of similar age and from the same geographical area. AB - BACKGROUND: Since the introduction of highly active antiretroviral therapy (HAART), the incidence of death in HIV-infected patients has dramatically decreased, and causes of death other than those related to HIV infection have increased, although it is unclear how these parameters compare with those in the age-matched general population living in the same geographical region. METHODS: Consecutive HIV-infected adults who were prescribed HAART in our hospital were prospectively followed from January 1997 to December 2004 or until death, loss to follow-up or discontinuation of HAART. Estimations of the annual incidence and causes of death in the general population of similar age in Catalonia per calendar year in the study period were obtained and compared with those in the HIV-infected cohort. RESULTS: There were 235 deaths among the 4471 patients on HAART (5%). The incidence of mortality decreased over time in HIV-infected patients (P<0.001; chi(2) test for trend), although it has remained approximately five times higher than that for the age-matched general population. AIDS-related events were the most common cause of death (n=95; 40%), although they significantly decreased over time (P<0.001; chi(2) test for trend), whereas liver diseases (P<0.001; chi(2) test for trend) and non-AIDS-defining infections (P=0.008; chi(2) test for trend) significantly increased over time. Infections in general (33 times higher), liver diseases (11 times higher) and non-Hodgkin lymphoma (5 times higher) were overrepresented as causes of death in the HIV infected cohort compared with the age-matched general population. CONCLUSIONS: Non-AIDS-defining infectious diseases, liver diseases, and non-Hodgkin lymphoma represent specific targets for efforts to further decrease mortality in HIV infected patients receiving HAART. PMID- 17461854 TI - Relative antiviral efficacy of ritonavir-boosted darunavir and ritonavir-boosted tipranavir vs. control protease inhibitor in the POWER and RESIST trials. AB - OBJECTIVE: To compare the relative antiviral efficacy of TMC114 with low-dose ritonavir (TMC114/r) and tipranavir with low-dose ritonavir (TPV/r) vs. control protease inhibitor (CPI) in treatment-experienced patients, using data from the POWER 1/2 and RESIST 1/2 trials. These trials recruited antiretroviral experienced patients with HIV RNA > 1000 HIV-1 RNA copies/mL and at least one primary PI mutation, and used optimized nucleoside reverse transcriptase inhibitors with or without enfuvirtide, plus investigator-selected CPI in the control arms. METHODS: For the POWER trials, data from the 600/100 mg twice a day (bid) dose and CPI arms (n=201) were included, while all data from the RESIST trials (TPV/r 500/200 mg bid and CPI; n=1159) were included. The difference in week 24 efficacy (intent to treat) for the new PI vs. CPI was compared between the trials. RESULTS: Overall baseline characteristics were well matched across the trials. At week 24, 72% of TMC114/r patients achieved a > or =1 log(10) copies/mL reduction in HIV RNA compared with 40% of TPV/r patients (for CPI patients, this percentage was 21 and 18%, respectively, in the POWER and RESIST trials). The treatment benefit of TMC114/r over CPI in the POWER trials was greater (outside the 95% confidence intervals) than the benefit of TPV/r over CPI in the RESIST trials, for the 24-week HIV RNA endpoints of 1 log(10) copies/mL reduction, <400 copies/mL and <50 copies/mL, and also for the mean rise in CD4 count. In sensitivity analysis, this difference in efficacy was strongest for those who did not also use enfuvirtide. CONCLUSIONS: Given the caveats of this type of analysis (for example, possible differences in trial conduct, and undetected differences in baseline resistance profiles), the efficacy benefits of TMC114/r vs. CPI in the POWER trials appear to be greater than the benefits of TPV/r vs. CPI in the RESIST trials, for patients who did not also use enfuvirtide. PMID- 17461855 TI - Seroprevalence of HIV infection in hospitalized paediatric patients at a tertiary care centre in western India. AB - The prevalence of HIV infection in children in India is not known. In this study, a total of 270 children admitted to paediatric wards in Mumbai were screened for HIV infection by enzyme-linked immunosorbent assay (ELISA). Eight patients (2.96%) had a positive HIV ELISA. Of these two patients (25%) were less than 15 months of old and hence HIV infection in them could not be confirmed. The prevalence of HIV was 2.3%. Three out of 11 patients with tuberculosis (27.3%) and four out of 15 patients with nutritional anaemia (26.7%) had HIV infection (P<0.0001 in each case). Vertical transmission was the cause in all children, suggesting that implementation of Prevention of Parent to Child Transmission of HIV (PPTCT) is required to reduce the rate of paediatric HIV infection. PMID- 17461856 TI - CD4 cell count and initiation of antiretroviral therapy: trends in seven UK centres, 1997-2003. AB - OBJECTIVES: We examined whether the timing of initiation of antiretroviral therapy (ART) in routine clinical practice reflected treatment guidelines, which have evolved towards recommending starting therapy at lower CD4 cell counts. METHODS: We analysed longitudinal data on 10,820 patients enrolled in the UK Collaborative HIV Cohort (UK CHIC) Study, which includes seven large clinical centres in south-east England. CD4 cell and viral load measurements performed in the period between 1 January 1997 and 31 December 2003 were classified according to whether ART was subsequently initiated or deferred, to estimate the probability of ART initiation by CD4 count and viral load over time. The effect of nonclinical factors (age, sex, ethnicity, and exposure category) was analysed by logistic regression. Kaplan-Meier analysis was used to estimate the proportion of patients who had initiated ART by a particular CD4 count among 'early' presenters (initial CD4 cell count >500 cells/microL). RESULTS: There was a tendency to initiate ART at lower CD4 cell counts over time in the years 1997 2000, especially in the range 200-500 cells/microL, with little change thereafter. An estimated 34% of HIV-infected individuals having presented early initiated ART at a CD4 count <200 cells/microL. We also found an independent influence of viral load, which was particularly pronounced for CD4 <350 cells/microL. Use of injection drugs was the only nonclinical factor associated with initiation of ART at lower CD4 cell counts. CONCLUSIONS: The initiation of ART in the clinics included in this analysis reflected evolving treatment guidelines. However, an unexpectedly high proportion of patients started ART at lower CD4 counts than recommended, which is only partly explained by late presentation. PMID- 17461857 TI - Antiviral activity of low-dose alovudine in antiretroviral-experienced patients: results from a 4-week randomized, double-blind, placebo-controlled dose-ranging trial. AB - BACKGROUND: Alovudine inhibits replication of highly nucleoside reverse transcriptase inhibitor (NRTI)-resistant HIV strains in vitro. However, dose dependent safety concerns resulted in its initial development being halted. Recently, a 4-week course of alovudine 7.5 mg/day added to a stavudine-free failing regimen yielded a significant decrease in viral load by -1.88 log(10) HIV 1 RNA copies/mL. The magnitude of the reduction in viral load suggested that lower doses might still be effective while offering adequate safety during long term use. OBJECTIVE: To determine whether lower dosages of alovudine still provide significant antiviral activity in patients with broad NRTI resistance. METHODS: A randomized, double-blind, placebo-controlled trial investigating three doses of alovudine (0.5, 1 and 2 mg) or placebo added for 4 weeks to a failing regimen in patients with evidence of NRTI-resistant HIV strains [>or=2 thymidine associated mutations (TAMs)]. The primary endpoint was the mean viral load reduction between baseline and week 4. RESULTS: Seventy-two patients were enrolled in the study: 21, 13, 18 and 20 in the placebo and 0.5, 1 and 2 mg arms, respectively. Baseline median CD4 count and viral load were 298 cells/microL (range 44-692 cells/microL) and 3.9 log(10) copies/mL (range 2.5-5.2 log(10) copies/mL), respectively. Baseline viral isolates harboured a median of four TAMs. Alovudine was added to a median four-drug failing regimen. At week 4, compared with placebo, mean viral load changes were -0.42 log(10) [95% confidence interval (CI) -0.67 to -0.18] and -0.30 log(10) (-0.55 to -0.06) in the 2 and 1 mg arms, respectively. There was no significant change in CD4 cell count. Alovudine was well tolerated. CONCLUSION: A 4-week course of alovudine 2 mg/day provided a modest but significant viral load reduction in patients harbouring viruses with a median of four TAMs. PMID- 17461858 TI - Susceptibility to opportunistic infections in HIV-infected patients with increased CD4 T-cell counts on antiretroviral therapy may be predicted by markers of dysfunctional effector memory CD4 T cells and B cells. AB - OBJECTIVES: HIV-infected patients responding to combination antiretroviral therapy (ART) after experiencing severe immunodeficiency may exhibit persistent immune defects and occasionally experience opportunistic infections (OIs) despite increased CD4 T-cell counts. The investigation of immune defects in such patients was examined in this study. METHODS: CD4 effector memory T-cell (T(em)-cell) function [assessed by blood cytomegalovirus (CMV) interferon-gamma (IFN-gamma) enzyme-linked immunosorbent spot-forming cell assay (ELISPOT) counts] and B-cell dysregulation [assessed by serum immunoglobulin A (IgA) and IgE levels] were examined in 27 patients with increased CD4 T-cell counts after receiving ART for over 2 years. Two of these patients and one other had developed OIs on ART and are described in detail. RESULTS: Serum levels of IgA and IgE were higher than reference intervals (P<0.001) and CMV IFN-gamma ELISPOT counts were lower than those in non-HIV-infected controls (P<0.001) in the HIV-infected patients. Low CMV IFN-gamma ELISPOT counts were associated with high IgA levels (r=-0.5, P=0.01, Spearman's correlation test) and segregated with high IgE levels (P=0.06, Fisher's test). CMV IFN-gamma ELISPOT counts and serum IgA and IgE levels did not change significantly over a median time of 35 (range 8-60) months after the first measurement, whereas CD4 T-cell counts increased. All three patients who experienced OIs had repeatedly low CMV IFN-gamma ELISPOT counts and increased serum levels of IgA and/or IgE. CONCLUSION: Low CD4 T(em)-cell function and B cell dysregulation are immune defects that may persist independently of changes in the CD4 T-cell count in HIV-1-infected patients responding to ART and are associated with an increased risk of developing an OI. PMID- 17461859 TI - Long-term evolution of CD4 count in patients with a plasma HIV RNA persistently <500 copies/mL during treatment with antiretroviral drugs. AB - BACKGROUND: The increase in CD4 count may reach a plateau after some duration of virological response to highly active antiretroviral therapy (HAART). METHODS: A total of 1281 HIV-infected patients initiating HAART were enrolled in the AntiPROtease (APROCO) cohort. We investigated determinants of increase in CD4 count using longitudinal mixed models in patients who maintained a plasma HIV RNA <500 HIV-1 RNA copies/mL. RESULTS: A total of 870 patients had a virological response at month 4. The median follow-up time was 57 months. Mean estimated increases in CD4 count in patients with persistent virological response were 29.9 cells/muL/month before month 4, 6.4 cells/microL/month between months 4 and 36, and 0.7 cells/microL/month (not significantly different from 0) after month 36. Three factors were associated with a significantly positive CD4 count slope after month 36: male gender (+0.9), no history of antiretroviral therapy at baseline (+1.7) and baseline CD4 count <100 cells/microL (+2.6). In patients who maintained a virological response after 5 years of HAART, a CD4 count >500 cells/microL was achieved in 83% of those with a baseline CD4 count >or=200 cells/microL and in 45% of those with a baseline CD4 count <200 cells/microL. CONCLUSION: The increase in CD4 count reaches a plateau after 3 years of virological response. Even if patients initiating HAART with low CD4 counts still show a CD4 count increase after 3 years, it remains insufficient to overcome immune deficiency in all patients. PMID- 17461860 TI - Characteristics and response to antiretroviral therapy of HIV-1-infected patients born in Africa and living in France. AB - BACKGROUND: The world-wide AIDS epidemic is reflected in Western Europe in an increasing number of HIV-infected persons who originate from Africa. We describe the characteristics and response to antiretroviral therapy (ART) of HIV-infected patients born in Africa and living in France. METHODS: Analysis of data from the (Anti PROtease COhorte APROCO) cohort study of HIV-infected patients initiating ART was carried out. Included in the study were 90 patients born in sub-Saharan Africa, 53 in North Africa and 771 in metropolitan France. RESULTS: At baseline, there was a higher proportion of women and of the heterosexual transmission route of infection among patients born in sub-Saharan Africa, a higher proportion of injecting drug users among patients born in North Africa and a higher frequency of unemployment and of unstable housing conditions among patients born in both sub-Saharan and North Africa as compared with patients born in France. The median CD4 cell count was lower in patients born in both sub-Saharan and North Africa (sub-Saharan Africa: 197 cells/microL; North Africa: 222 cells/microL) than in patients born in France (307 cells/microL). Median HIV-1 viral loads were similar. After a median follow-up time of 36 months (2506 patient-years), the Kaplan-Meier estimations of probability of survival without new AIDS-defining events were not different. After 36 months of ART, in multivariate analysis, median CD4 cell count, CD4/CD8 ratio and viral load were not statistically different according to birthplace, but the median CD4 percentage was lower in patients born in both sub-Saharan and North Africa. The adherence profiles were similar. CONCLUSIONS: Although clinical response and adherence to ART did not appear to differ in patients according to their birthplace, the reasons for the more advanced HIV infection observed at ART initiation among patients born in Africa should be further investigated. PMID- 17461861 TI - Combination antiretroviral therapy without a nucleoside reverse transcriptase inhibitor: experience from 334 patients in three cohorts. AB - BACKGROUND: Toxicity and resistance may limit the use of HIV nucleoside reverse transcriptase inhibitors (NRTIs). We assessed the safety and activity of regimens that did not include an NRTI. METHOD AND PATIENTS: We analysed NRTI-sparing regimens using pooled data from three cohorts in Australia and France where HIV RNA viral load, CD4 lymphocyte count and metabolic parameters are assessed prospectively. The inclusion criterion was the commencement of any antiretroviral combination excluding NRTIs. RESULTS: A total of 334 (3.9%) of 8477 patients were included in the present study for a median follow-up time of 105 weeks. Therapeutic combinations were one nonnucleoside reverse transcriptase inhibitor (NNRTI) plus one protease inhibitor (PI) (58%), two PIs (26%), one PI (16%), and one NNRTI plus two PIs (8%). At baseline, the median CD4 lymphocyte count was 264 cells/muL (interquartile range 164-446 cells/muL) and 25% of patients had plasma HIV RNA below 500 HIV-1 RNA copies/mL. In intent-to-treat analysis, 64% of patients had HIV RNA <500 copies/mL at 6 months and 68% at 24 months. The mean CD4 lymphocyte count increase was 60 cells/microL (95% confidence interval 41-76 cells/microL) at 6 months and 111 cells/microL (95% confidence interval 82-140 cells/microL) at 24 months. Prognostic factors for having HIV RNA <500 copies/mL at 6 months included independently having undetectable HIV RNA at baseline and being naive for NNRTIs. The proportion of patients with triglycerides >2.3 mmol/L increased from 32% to 63% at 6 months and to 62% at 24 months (P-trend=0.002), and those with total cholesterol >6.2 mmol/L increased from 18% to 38% at 6 months and to 44% at 24 months (P-trend <0.001), with an increased risk for patients treated with NNRTI+PIs. Forty-one per cent of patients discontinued their NRTI-sparing regimen. CONCLUSION: In these antiretroviral-experienced patients, NRTI-sparing therapy appeared to have satisfactory virological and immunological efficacy. However, hyperlipidaemia was frequent and requires monitoring of cardiovascular risk factors. PMID- 17461862 TI - Setting a minimum threshold CD4 count for initiation of highly active antiretroviral therapy in HIV-infected patients. AB - The aim of our study was to determine a minimum threshold CD4 count for highly active antiretroviral therapy (HAART) initiation in HIV-infected patients. A schema using longitudinal data from a clinical cohort was designed. The presenting CD4 counts of asymptomatic HIV-infected patients in Hong Kong were evaluated in relation to their progression to AIDS within 1 year of diagnosis of HIV infection. A graph was generated to depict the changes in the percentage of cumulative AIDS diagnoses for every 10 cell/microL increase in presenting CD4 count. Of 181 patients, 24 had developed AIDS within 1 year of diagnosis of HIV infection. Setting the CD4 count threshold at 150 cells/microL gave a good balance between the number of preventable AIDS-defining events and the number of non-AIDS patients initiating HAART. No extra AIDS-defining events occurred when the CD4 count threshold was reduced from 200 to 150 cells/microL, despite the addition of 13 more patients. In multivariate Cox regression analysis, presenting CD4 count was a significant predictor for AIDS occurrence. The relative hazard for AIDS occurrence of patients with presenting CD4 counts 6 months) were reprogrammed to VVI 40 to allow for native conduction to resume. After 5 minutes of native rhythm, a surface electrocardiogram (ECG) was recorded, and then the previous device settings were restored. This ECG was compared to the preimplant ECG. Preimplant mean ejection fraction was 19% (range, 10%-35%), and follow-up mean ejection fraction was 35% (12.5%-65%). Mean time from implant to follow-up ECG was 14 months (range, 6-31). The QRS interval prior to CRT was 155 +/- 29 ms, and shortened to 144 +/- 31 ms (P = 0.0006), and the QRS axis shifted from -1 +/- 59 to -26 +/- 53 (P = 0.03). There was no significant change in PR or QTc interval, or in heart rate. CONCLUSION: CRT leads to a decrease in the surface QRS duration, without affecting other surface ECG parameters. The reduced electrical activation time may reflect changes in the specialized conduction system or in intramyocardial impulse transmission. PMID- 17461867 TI - Achieving sufficient safety margins with fixed duration waveforms and the use of multiple time constants. AB - INTRODUCTION: There are several options to achieve a sufficient safety margin in a patient with a high defibrillation threshold (DFT), with varying and typically modest success. Programming fixed (millisecond) durations of both phases of a biphasic waveform in an implantable cardioverter defibrillator (ICD) has demonstrated utility. METHODS: We established an informal multisite registry of ICD implanting facilities. Each facility agreed to attempt the use of fixed duration waveforms whenever there was an inadequate safety margin with tilt-based waveforms. A 3.5-ms-based fixed duration shock was tried first. If that failed to achieve a 10-J safety margin then a 2-ms-based shock was used. We also tabulated an HEDFT (high estimate DFT) as precise DFTs were not determined. RESULTS: Sixteen patients (15 M, 1 F) were entered into the registry (age 58.2 +/- 17.9 years) with ejection fractions of .30 +/-.11. Superior vena cava coils were used in 7 patients according to physician preference. The tilt-based HEDFTs were 35.4 +/- 3.2 J delivered and 35.8 +/- 3.3 J stored energy. The 3.5-ms based shocks were evaluated on 14 patients and the HEDFT fell to 23.4 +/- 6.3 J delivered (P < 0.0001) and 26.2 +/- 6.9 J stored energy (P < 0.0001). The 2-ms-based fixed duration shocks were then evaluated on 6 patients and the delivered energy HEDFT was 22.2 +/- 5.8 J (P = 0.001 vs. tilt-based shocks) while the stored energy HEDFT was 27.9 +/- 6.4 J (P = 0.01 vs. tilt-based shocks). Using the better of the two fixed duration waveforms, the mean safety margin was improved from -1.2 +/- 1.9 J to 9.5 +/- 5.9 J (P < 0.00001). Multivariate predictors of the safety margin improvement were the absence of the Superior Vena Cava (SVC) coil and absence of Ventricular fibrillation (VF) presentation. Four patients still required lead repositioning after the use of the fixed duration waveforms. No additional leads were implanted. CONCLUSION: The use of a selection of directly programmed fixed duration biphasic shocks had a striking impact on the HEDFT for these difficult patients. Adequate safety margins were obtained for 12 of 16 patients with no lead manipulation or other approaches. PMID- 17461868 TI - Elevated estimated pulmonary artery systolic pressure is associated with an adverse clinical outcome in patients receiving cardiac resynchronization therapy. AB - BACKGROUND: A substantial percentage of patients with heart failure remain nonresponsive to cardiac resynchronization therapy (CRT). There is a paucity of information on the impact of baseline elevated pulmonary artery pressure on clinical outcome and on left ventricular reverse remodeling (LV-RR) after CRT. We sought to investigate the impact of elevated estimated pulmonary artery systolic pressure (ePASP) on clinical outcome and LV-RR after CRT. METHODS: This study retrospectively analyzed data from 68 subjects with standard indications for CRT over a 12-month period. Subjects were stratified into two groups based on the echocardiographic estimation of pulmonary artery pressure i.e., ePASP > or = 50 mmHg (n = 27) and ePASP < 50 mmHg (n = 41). Long-term response was measured as a combined endpoint of heart failure hospitalizations and all cause mortality at 12 months, and compared within the two groups using the Kaplan-Meier method. Follow up echocardiograms to assess for LV-RR were available in 51 subjects (mean duration 7.1 months). LV-RR was defined as any improvement in global systolic function with reduction in left ventricular internal diameter. RESULTS: The study population was composed of 24 women and 44 men (age, mean +/- SD; 70 +/- 11 years), with a decreased left ventricular ejection fraction ([25 +/- 9]%) and a wide QRS (171 +/- 54 ms). There were no significant differences in the clinical features between the high and low ePASP group. Subjects with ePASP > or = 50 mmHg had a significantly worse clinical outcome (Hazard ratio (95% CI), 2.0 (1.2-5.5), P = 0.02). Baseline ePASP was not predictive of LV-RR (P = 0.32). CONCLUSION: In patients receiving CRT, although elevated estimated pulmonary artery systolic pressure (ePASP > or = 50 mmHg) does not significantly impact LV reverse remodeling, it is associated with an adverse long-term outcome. PMID- 17461869 TI - Sedation with midazolam for electrical cardioversion. AB - BACKGROUND: Electrical cardioversion (ECV) usually requires the assistance of the anesthesiology team. To avoid this dependence, previous studies have considered the use of sedation with benzodiazepines administered by cardiologists. We describe our experience with intravenous Midazolam during cardioversion. METHODS: We performed 280 ECV in 202 patients sedated with intravenous Midazolam, without anesthesiology supervision. In scheduled cardioversions, we tested two protocols of Midazolam administration: a bolus of 3 mg, followed by 2 mg each minute until necessary, and a loading dose of 0.09-0.1 mg/kg. In cardioversions performed during electrophysiology studies or defibrillator implant, Midazolam was administered by small repeated doses during the entire procedure. RESULTS: Midazolam was effective to obtain adequate sedation in 99% of cases. All patients had amnesia with regards of the cardioversion. A loading dose of Midazolam allowed a shortening of the procedural time without serious adverse events. Intubation or the assistance of an anesthetist was never necessary. CONCLUSION: Sedation with Midazolam for ECV is effective and well tolerated, with some cautions discussed. A loading dose of Midazolam is well tolerated and further reduces the procedural time. PMID- 17461870 TI - Strength duration curve for epicardial left ventricular stimulation. AB - BACKGROUND: The strength duration curve for endocardial stimulation has been extensively studied. Little information is available on the left ventricular epicardial strength duration curve. In view of the large number of patients treated with resynchronization therapy, left ventricular epicardial stimulation parameters have practical importance. METHODS: Twelve patients who underwent implant of a biventricular defibrillator were available for at least 4 months of follow up and accurate determination of strength duration curves were studied. Strength duration curves were constructed at 30 days (subacute phase) and 4 or more months (chronic phase) after the implant for right ventricular endocardial, left ventricular epicardial unipolar, and left ventricular bipolar stimulation. The goal was to determine the chronaxie, which correlates with the most economical stimulation. RESULTS: There was no significant difference between the right ventricular endocardial and left ventricular epicardial bipolar chronaxie (P = 0.57 subacute and 0.6 chronic) or right ventricular endocardial and left ventricular unipolar chronaxie (P = 0.93 subacute and 0.92 chronic). Most chronaxie values were lower than the factory default values. CONCLUSION: The left ventricular unipolar or bipolar epicardial chronaxie is not significantly different from the right ventricular endocardial chronaxie. Both values are lower than the pulse duration used as default setting in most devices as well as in clinical practice. Individual determination of the chronaxie could lead to energy savings. PMID- 17461871 TI - Prevalence of mechanical dyssynchrony in heart failure patients with different QRS durations. AB - BACKGROUND: Cardiac resynchronization therapy (CRT) has emerged as an established therapy for congestive heart failure. However, up to 30% of patients fail to respond to CRT despite prolonged QRS. OBJECTIVES: This study aimed at defining the prevalence of interventricular and intraventricular dyssynchrony in heart failure patients with different QRS durations. METHODS: A total of 123 consecutive patients with severe heart failure (LVEF < 35% and NYHA class III-IV) were prospectively evaluated using 12-lead electrocardiogram and complete echocardiographic examination including tissue Doppler imaging. RESULTS: According to the QRS duration, 56 patients had a QRS duration < or = 120 ms (Group 1), 33 patients had a QRS duration between 120 and 150 ms (Group 2), and 34 patients had a QRS duration > or = 150 ms (Group 3). Intraventricular dyssynchrony was present in 36% of Group 1 patients, in 58% of Group 2 patients, and in 79% of Group 3 patients (P < 0.000). Linear regression demonstrated a weak relation between QRS and intraventricular dyssynchrony. A greater proportion of patients with interventricular dyssynchrony was observed in Group 3 or Group 2 compared to patients with normal QRS duration (32% in Group 1 vs. 51.5% in Group 2 vs. 76.5% in Group 3, P < 0.000). Linear regression demonstrated a significant relation between QRS duration and interventricular mechanical delay. CONCLUSIONS: Although both interventricular and intraventricular dyssynchrony increased with the increasing QRS duration, the correlation between intraventricular mechanical and electrical dyssynchrony was weak. The lack of intraventricular dyssynchrony in a fraction of patients with standard CRT indication by QRS duration may provide us insight into the nonresponders rates. PMID- 17461872 TI - Investigation of various types of neurocardiogenic response to head-up tilting by extended hemodynamic and neurohumoral monitoring. AB - BACKGROUND: The pathophysiology of neurocardiogenic syncope is heterogeneous. This study aim was to analyze whether extended monitoring during tilt-table testing provided additional information on the hemodynamic and vegetative state prior to neurocardiogenic syncope. METHODS: This retrospective analysis is based on data of head-up tilt-table testing of 40 unselected consecutive patients with a history of unexplained syncope. For optimized characterization of the type of syncope, monitoring included electrocardiogram (ECG), blood pressure measurements and cardiac output, peripheral vascular resistance and contractility index measurements by impedance cardiography, as well as epinephrine and norepinephrine plasma levels in supine position and every 5 min during tilting. RESULTS: Seven of 40 patients were unsuitable for analysis because of incomplete data sets. Tilt table was positive in 26 patients, negative in 7. Groups did not differ in hemodynamic and catecholaminergic parameters at baseline. Responses to tilting were VASIS 1 (mix of cardioinhibitory and vasodepressor) in 5 patients, VASIS 2B (cardioinhibitory with asystole >3 sec) in 3, VASIS 3 (vasodepressor) in 16, orthostatic dysregulation in 2. In VASIS 1, the catecholamine measurement 4 min before syncope showed a proportionally larger increase of the epinephrine level than of norepinephrine. This disproportion was not observed in VASIS 2B and 3. In VASIS 2B, strong vasoconstriction and negative inotropy were evident in the presyncopal period. In VASIS 3, vascular resistance decreased continuously before syncope, while contractility index increased inadequately. Presyncopal epinephrine surge or norepinephrine loss was not observed in this group, suspecting other vasodilating factors. CONCLUSIONS: Extended monitoring by impedance cardiography and plasma catecholamine measurements during tilt-table testing gave further insight into different hemodynamic and neurohumoral presyncopal patterns among the various types of neurocardiogenic syncope and may thereby help to develop individualized therapeutic concepts. PMID- 17461873 TI - Effects of intracoronary low-dose enalaprilat on ventricular repolarization dynamics after direct percutaneous intervention for acute myocardial infarction. AB - BACKGROUND: Data from animal models suggest that inhibition of angiotensin converting enzymes result in an increased ventricular electrical stability after reperfusion in acute myocardial infarction (MI). As electrical stability is largely dependent on ventricular repolarization, we sought to determine the impact of low-dose intracoronary (i.c.) application of enalaprilat (EN) as an adjunct to direct primary coronary intervention (PCI) on QT dynamics in the acute phase of MI. METHODS: Twenty-two consecutive patients with a first acute MI who underwent successful direct PCI (TIMI 3 flow) were randomized to i.c. EN (50 microg) or placebo/saline (PL), given immediately after reopening of the infarct vessel. On hospital admission, a 24-hour-Holter-electrocardiogram (ECG) was initiated. Slopes of the linear QT/RR regression were determined for the time intervals before reperfusion and after reperfusion. RESULTS: A total of 7 patients in the EN group and 8 patients in the PL group had valid ECG recordings for beat-to-beat QT analysis. Mean RR interval and mean QT interval were not significantly different between the EN and the PL groups both before and after PCI. There were also no significant differences regarding QT/RR slopes between EN and PL groups before PCI. After PCI, QT/RR slopes significantly decreased in the EN group (0.169 +/- 0.04 to 0.121 +/- 0.03; P < 0.01), whereas there were no significant alterations in the PL group (0.175 +/- 0.04 to 0.171 +/- 0.03; P = ns). CONCLUSIONS: Intracoronary EN therapy as an adjunct to direct PCI significantly decreases QT/RR slopes, suggesting a normalization of the coupling between heart rate and repolarization by improving electrical restitution. Thus, our findings offer new insights into possible beneficial effects of ACE inhibition on cardiac electrical stability in acute MI. PMID- 17461874 TI - Is home orthostatic self-training effective in preventing neurally mediated syncope? AB - BACKGROUND: Repeated orthostatic stress may prove to be of benefit in the regulation of neurally mediated syncope. But the role of home orthostatic self training is not established to prevent symptoms in patients with neurally mediated syncope. We performed a prospective and randomized study to evaluate the effectiveness of repeated home orthostatic self-training in preventing tilt induced neurally mediated syncope. METHODS AND RESULTS: Fourty-two consecutive patients (24 males and 18 females, mean age 39 years, 16-68 years) with recurrent neurally mediated syncope were randomized into the tilt training and control groups. The home orthostatic self-training program consisted of daily sessions for 7 days a week for 4 weeks. In order to determine the effects of home orthostatic self-training, we repeated the head-up tilt test in both groups 4 weeks later. Among the tilt-training group, 9 of 16 patients (56%) had a positive response on follow-up head-up tilt test. Among the untreated control group, 9 of 17 patients (53%) had a positive response on follow-up head-up tilt test. In subgroup analyses according to the number of tilt-training sessions or the classified type, we found no differences in the follow-up head-up tilt test responses. Spontaneous syncope or presyncope over mean follow-up of 16.9 months were observed in 42.9% versus 47.1% in the tilt-training and control group, respectively. CONCLUSIONS: Home orthostatic self-training was ineffective in reducing the positive response rate of head-up tilt test in patients with recurrent neurally mediated syncope. PMID- 17461875 TI - Determinants of lesion sizes and tissue temperatures during catheter cryoablation. AB - BACKGROUND: Factors which influence lesion size from catheter-based cryoablation have not been well described. This study describes factors which influence lesion size during catheter cryoablation. METHODS AND RESULTS: Cryoablation was delivered to porcine left ventricular myocardium in a saline bath using 4- or 8 mm electrode catheters. Ablation was delivered with the electrodes either vertical or horizontal to the tissue and both with and without superfusate flow over the electrode. The effect of electrode contact pressure was tested. Lesion dimensions were measured. All experiments were duplicated to measure tissue temperatures at 1-, 2-, 3-, and 5-mm deep to the ablation electrode. The 8-mm electrode produced lower tissue temperatures and larger lesion volumes when compared with the 4-mm electrode (all P < 0.05). Superfusate flow slowed the rate of tissue cooling, markedly warmed tissue temperatures, and reduced lesion volume when compared with no flow conditions. By linear regression modeling, lesion sizes and tissue temperatures were related to the presence of superfusate flow, electrode orientation, contact pressure and electrode size, or catheter refrigerant flow rate (r2 for models = 0.90-0.96, all P < 0.001). Electrode temperature predicted lesion size or tissue temperatures only when analyzed independent of electrode size or refrigerant flow rate. CONCLUSIONS: Lesion sizes and tissue temperatures during catheter cryoablation are related to convective warming, electrode orientation, electrode contact pressure, and any of the following: electrode size, catheter refrigerant flow rate or electrode temperature. However, electrode temperature may be a poor predictor of lesion size and tissue temperature for a given catheter size. PMID- 17461876 TI - The results of radiofrequency catheter ablation of supraventricular tachycardia in children. AB - BACKGROUND: Radiofrequency (RF) catheter ablation represents a major advance in the management of children with cardiac arrhythmias and has rapidly become the standard of care for the first-ling therapy of supraventricular tachycardias (SVTs). The purpose of this study was to investigate the results of the RF catheter ablation of SVTs in pediatric patients. METHODS: From December 1989 to August 2005, a total of 228 pediatric patients (age: 9 +/- 7 years, range: 5-18 years; male:female = 117:111) with clinically documented SVT underwent an electrophysiologic study and RF catheter ablation at our institution. RESULTS: The arrhythmias included atrioventricular reentrant tachycardia (AVRT; n = 140, 61%), atrioventricular nodal reentrant tachycardia (AVNRT; n = 66, 29%), atrial tachycardia (AT; n = 11, 5%), and atrial flutter (AFL; n = 11, 5%). The success rate of the RF catheter ablation was 92% for AVRT, 97% for AVNRT, 82% for AT, and 91% for AFL, respectively. Procedure-related complications were infrequent (8.7%; major complications: high grade AV block (2/231, 0.9%); minor complications: first degree AV block (6/231, 2.6%), reversible brachial plexus injury (2/231, 0.9%), and local hematomas or bruises (10/231, 4.3%)). The recurrence rate was 4.7% (10/212) during a follow-up period of 86 +/- 38 months (0.5-185 months). CONCLUSIONS: The RF catheter ablation was a safe and effective method to manage children with paroxysmal and incessant tachycardia. The substrates of the arrhythmias differed between the pediatric and adult patients. However, the success rate of the ablation, complications, and recurrence during childhood were similar to those of adults. PMID- 17461877 TI - Incidence and initial characteristics of pilsicainide-induced ventricular arrhythmias in patients with Brugada syndrome. AB - BACKGROUND: In patients with Brugada syndrome, class I antiarrhythmic drugs can trigger ventricular arrhythmias (VA). The incidence and initial characteristics of VA that developed after pilsicainide was examined in 28 patients with Brugada type electrocardiographic (ECG) abnormalities and with a positive response in the pilsicainide test. The clinical outcome was also compared between patients with and without pilsicainide-induced VA. METHODS AND RESULTS: In all patients, pilsicainide increased ST segment elevation and accentuated type 1 ECG changes. Ventricular tachycardia (VT) developed in 3 patients and premature ventricular complexes (PVC) in 2 other patients. These 5 patients (group I) had higher ST segment elevation in lead V2 on the ECG at baseline and after pilsicainide and showed a longer QTc interval after pilsicainide than the other 23 patients (group II). However, there was no difference between the 2 groups regarding incidence of prior cardiac events, results of signal-averaged ECG, HV interval, inducibility of ventricular fibrillation by programmed electrical stimulation, or QRS duration. In 1 patient, PVC originated from 3 sites, 2 of which triggered polymorphic VT. The right ventricular (RV) outflow tract was the origin of 2 types of PVC, and other RV sites of 5 other types. During a 45 +/- 37 months follow-up, polymorphic VT recurred in 2 patients in group II. CONCLUSIONS: Pilsicainide induced VA in some patients with Brugada syndrome, but this result may not be used as a parameter of the risk stratification of Brugada syndrome. Multiple PVC induced by pilsicainide and triggering polymorphic VT originated from several RV sites is an important factor when considering patients for treatment with catheter ablation. PMID- 17461878 TI - Post Glenn shunt: how to pace? PMID- 17461879 TI - The dilemma of ICD implant testing. AB - Ventricular fibrillation (VF) has been induced at implantable cardioverter defibrillator (ICD) implant to ensure reliable sensing, detection, and defibrillation. Despite its risks, the value was self-evident for early ICDs: failure of defibrillation was common, recipients had a high risk of ventricular tachycardia (VT) or VF, and the only therapy for rapid VT or VF was a shock. Today, failure of defibrillation is rare, the risk of VT/VF is lower in some recipients, antitachycardia pacing is applied for fast VT, and vulnerability testing permits assessment of defibrillation efficacy without inducing VF in most patients. This review reappraises ICD implant testing. At implant, defibrillation success is influenced by both predictable and unpredictable factors, including those related to the patient, ICD system, drugs, and complications. For left pectoral implants of high-output ICDs, the probability of passing a 10 J safety margin is approximately 95%, the probability that a maximum output shock will defibrillate is approximately 99%, and the incidence of system revision based on testing is < or = 5%. Bayes' Theorem predicts that implant testing identifies < or = 50% of patients at high risk for unsuccessful defibrillation. Most patients who fail implant criteria have false negative tests and may undergo unnecessary revision of their ICD systems. The first-shock success rate for spontaneous VT/VF ranges from 83% to 93%, lower than that for induced VF. Thus, shocks for spontaneous VT/VF fail for reasons that are not evaluated at implant. Whether system revision based on implant testing improves this success rate is unknown. The risks of implant testing include those related to VF and those related to shocks alone. The former may be due to circulatory arrest alone or the combination of circulatory arrest and shocks. Vulnerability testing reduces risks related to VF, but not those related to shocks. Mortality from implant testing probably is 0.1-0.2%. Overall, VF should be induced to assess sensing in approximately 5% of ICD recipients. Defibrillation or vulnerability testing is indicated in 20-40% of recipients who can be identified as having a higher-than usual probability of an inadequate defibrillation safety margin based on patient specific factors. However, implant testing is too risky in approximately 5% of recipients and may not be worth the risks in 10-30%. In 25-50% of ICD recipients, testing cannot be identified as either critical or contraindicated. PMID- 17461880 TI - Idiopathic ventricular fibrillation successfully terminated by radiofrequency ablation of the distal Purkinje fibers. AB - A 21-year-old woman was admitted for evaluation of recurrent episodes of syncope. She had several spontaneous and abrupt loss of consciousness episodes while at the emergency department, caused by sustained and nonsustained ventricular fibrillation episodes. Each episode was initiated by a certain premature ventricular complex, which remained the same in subsequent ventricular fibrillation episodes. She had a total of eight more episodes of ventricular fibrillation during her admission, despite administration of intravenous antiarrhythmic agents. A diagnosis of idiopathic ventricular fibrillation was made. Radiofrequency catheter ablation was performed, targeting the distal Purkinje system. Ventricular fibrillation was noninducible after the procedure, and the patient has been symptom-free for the past 1 year. PMID- 17461881 TI - Early pacemaker battery depletion caused by a current leak in the output circuitry: rectification not exchange. AB - Malfunction of a dual chamber system pacemaker due to a current leak in the output circuitry leading to a rapid decline in battery longevity is described. Serial pulse amplitude alterations in defined device configurations enabled location of the defect in the 2.5-V output voltage level; the current leak disappeared in the 5-V output voltage level, and recurred in the 7.5-V output voltage level which combines both 2.5-V and 5-V output circuitries. Reprogramming of pulse amplitudes rectified device malfunction. Circuitry reprogramming of pacemakers with faulty unexplained high battery current can circumvent early replacement in management of device malfunction. PMID- 17461882 TI - A very narrow preexisting isthmus in a case with typical atrial flutter. AB - A 61-year-old woman with typical atrial flutter underwent an electrophysiologic study and radiofrequency catheter ablation. The electroanatomic mapping revealed two contiguous lines of distinct double potentials (DPs) extending anteriorly/posteriorly from the coronary sinus ostium to the inferior vena cava (IVC) border. A large part of the anterior line of the DPs was close and parallel to the tricuspid annulus (TA). An initial discrete radiofrequency application at the very narrow preexisting isthmus between the TA and anterior line of the DPs completed the IVC-TA isthmus conduction block. PMID- 17461883 TI - Brugada pattern electrocardiogram associated with supratherapeutic phenytoin levels and the risk of sudden death. AB - The emergence of Brugada pattern on electrocardiogram in response to class IA or IC antiarrhythmic agents is widely utilized to diagnose concealed Brugada syndrome and recognized as a risk factor for sudden death. Phenytoin, a class IB antiarrhythmic agent, has not been reported to induce Brugada pattern. We report a patient who presented with Brugada electrocardiogram at supratherapeutic phenytoin level. Considering that patients with syncope may falsely be labeled to have seizures and some epilepsy patients are at increased risk of sudden death, all patients with supratherapeutic phenytoin level should be evaluated with an electrocardiogram for emergence of Brugada pattern. PMID- 17461884 TI - Runaway implantable defibrillator--a rare complication of radiation therapy. AB - A case of a patient with runaway implantable cardioverter defibrillator (ICD) due to radiation therapy of a lung cancer is reported. This manifested as poorly tolerated wide complex tachycardia due to inappropriate rapid ventricular pacing, The event terminated with polymorphic VT, which inhibited pacing and ceased spontaneously before ICD discharge. The likely cause was corruption of device random access memory by ionizing radiation. PMID- 17461885 TI - An unusual case of the cephalic vein with a supraclavicular course. AB - While the subclavian or axillary vein can be safely and successfully punctured in the majority of cases, some device implanters still prefer cut down to the cephalic vein as the initial approach to venous access for transvenous placement of pacemaker or defibrillator leads out of concern for the risk of pneumothorax, subclavian crush, and other possible complications. However, very occasionally, the cephalic vein crosses superficial to the clavicle to join the external jugular vein, making it rather unappealing for this purpose. Relying on a guide wire introduced through the cephalic vein to guide puncture of the subclavian vein is unlikely to be successful in such a situation and may cause accidental damage to the vital structures in the thoracic inlet region. Device implanters need to be aware of this anatomical anomaly because of the implications for transvenous lead placement. PMID- 17461887 TI - MD-354: what is it good for? AB - MD-354 (meta-chlorophenylguanidine) has been identified as a member of a novel class of 5-HT(3) serotonin receptor agonists. MD-354 is a 5-HT(3) receptor partial agonist that has been shown to behave as an agonist in some assays, and as an antagonist in others. MD-354 also binds at alpha-adrenoceptors (ARs) and displays an affinity for alpha(2B)-ARs comparable to its affinity for 5-HT(3) receptors. Although devoid of antinociceptive actions following systemic administration alone, MD-354 markedly enhances the antinociceptive actions of clonidine in the mouse tail-flick assay without potentiating the sedative side effects of clonidine. Although studies with MD-354 are still in progress, some pharmacological findings are described here. MD-354-related agents may represent drug adjuvants for the relief of severe pain. PMID- 17461888 TI - Lacosamide: a review of preclinical properties. AB - Lacosamide (LCM), (SPM 927, (R)-2-acetamido-N-benzyl-3-methoxypropionamide, previously referred to as harkoseride or ADD 234037) is a member of a series of functionalized amino acids that were specifically synthesized as anticonvulsive drug candidates. LCM has demonstrated antiepileptic effectiveness in different rodent seizure models and antinociceptive potential in experimental animal models that reflect distinct types and symptoms of neuropathic as well as chronic inflammatory pain. Recent results suggest that LCM has a dual mode of action underlying its anticonvulsant and analgesic activity. It was found that LCM selectively enhances slow inactivation of voltage-gated sodium channels without affecting fast inactivation. Furthermore, employing proteomic affinity-labeling techniques, collapsin-response mediator protein 2 (CRMP-2 alias DRP-2) was identified as a binding partner. Follow-up experiments confirmed a functional interaction of LCM with CRMP-2 in vitro. LCM did not inhibit or induce a wide variety of cytochrome P450 enzymes at therapeutic concentrations. In safety pharmacology and toxicology studies conducted in mice, rats, rabbits, and dogs, LCM was well tolerated. Either none or only minor side effects were observed in safety studies involving the central nervous, respiratory, gastrointestinal, and renal systems and there is no indication of abuse liability. Repeated dose toxicity studies demonstrated that after either intravenous or oral administration of LCM the adverse events were reversible and consisted mostly of exaggerated pharmacodynamic effects on the CNS. No genotoxic or carcinogenic effects were observed in vivo, and LCM showed a favorable profile in reproductive and developmental animal studies. Currently, LCM is in a late stage of clinical development as an adjunctive treatment for patients with uncontrolled partial onset seizures, and it is being assessed as monotherapy in patients with painful diabetic neuropathy. Further trials to identify LCM's potential in pain and for other indications have been initiated. PMID- 17461889 TI - Levetiracetam: the profile of a novel anticonvulsant drug-part I: preclinical data. AB - The objective of this article was to review and summarize the available reports on the preclinical profile of the novel anticonvulsant drug levetiracetam (LEV). Therefore, a careful search was conducted in the MEDLINE database and combined with guidelines from regulatory agencies, proceedings of professional scientific meetings, and information provided by the manufacturers. This article provides detailed information on the anticonvulsant effects of LEV in various animal models of epilepsy and on its pharmacology in laboratory animals. The mechanism of action of LEV is reviewed, with special regard to its recently discovered binding site, the synaptic vesicle protein 2A. In general, LEV is shown to be a safe, broad-spectrum anticonvulsant drug with highly beneficial pharmacokinetic properties and a distinct mechanism of action. The clinical studies with LEV will be discussed in the second part of this review article to be published subsequently. PMID- 17461890 TI - Levetiracetam: part II, the clinical profile of a novel anticonvulsant drug. AB - The objective of this article was to review and summarize the available reports on the profile of the novel anticonvulsant drug levetiracetam (LEV) in a clinical setting. Therefore, a careful search was conducted in the MEDLINE database and combined with guidelines from regulatory agencies, proceedings of professional scientific meetings, and information provided by the manufacturers. This article is devoted to the clinical pharmacology and clinical trials of LEV investigating its efficacy and safety as add-on therapy or monotherapy for various seizure types. Finally, results from postmarketing surveillance of LEV are briefly discussed. In general, LEV is shown to be a safe, broad-spectrum anticonvulsant drug with highly beneficial pharmacokinetic properties, a favorable long-term retention rate, and a high responder rate, indicating that LEV is an efficient therapeutic option for the treatment of several types of epilepsy. PMID- 17461891 TI - Pharmacological properties, toxicology and scientific rationale for the use of natalizumab (Tysabri) in inflammatory diseases. AB - Natalizumab (Tysabri) was the first adhesion molecule antagonist to make it into clinical trial for patients with multiple sclerosis (MS) and other inflammatory disorders. Natalizumab is a humanized recombinant monoclonal antibody (MAb) that binds to the alpha (alpha)(4) chain of the alpha(4) beta (beta)(1) (very late activating antigen 4; VLA-4) and alpha(4)beta(7) integrins. The scientific rationale for natalizumab therapy is the reduction of leukocyte extravasation into peripheral tissues. Natalizumab, like other VLA-4 antagonists, may also interfere with the activation of T lymphocytes in secondary lymphoid organs and their reactivation in the central nervous system (CNS). Shortly after its approval for the treatment of relapsing-remitting MS (RR-MS), three patients who were treated with natalizumab in the setting of clinical trials developed progressive multifocal leukoencephalopathy (PML), an opportunistic infection of the brain with the polyoma virus JC. It remains to be elucidated why the use of this VLA-4 antagonist is associated with an increased incidence of PML. Natalizumab was recently reapproved for the treatment of relapsing forms of MS. In this review, we outline the scientific rationale for using natalizumab in MS and other inflammatory disorders. In addition, an overview of pharmacological properties, clinical efficacy, safety, and toxicology of natalizumab is provided. PMID- 17461892 TI - Dextromethorphan: a review of N-methyl-d-aspartate receptor antagonist in the management of pain. AB - Dextromethorphan (DM) is a noncompetitive N-methyl-d-aspartate (NMDA) receptor antagonist, which is widely used as an antitussive agent. DM also prevents neuronal damage and modulates pain sensation via noncompetitive antagonism of excitatory amino acids (EAAs). DM has been found to be useful in the treatment of pain in cancer patients and in the treatment of methotrexate-induced neurotoxicity. Clinical studies with DM in cancer patients are reviewed in this article. PMID- 17461893 TI - The pharmacology of Ro 64-6198, a systemically active, nonpeptide NOP receptor (opiate receptor-like 1, ORL-1) agonist with diverse preclinical therapeutic activity. AB - The NOP receptor (formerly referred to as opiate receptor-like 1, ORL-1, LC132, OP(4), or NOP(1)) is a G protein-coupled receptor that shares high homology to the classic opioid MOP, DOP, and KOP (mu, delta, and kappa, respectively) receptors and was first cloned in 1994 by several groups. The NOP receptor remained an orphan receptor until 1995, when the endogenous neuropeptide agonist, known as nociceptin or orphanin FQ (N/OFQ) was isolated. Five years later, a group at Hoffmann-La Roche reported on the selective, nonpeptide NOP agonist Ro 64-6198, which became the most extensively published nonpeptide NOP agonist and a valuable pharmacological tool in determining the potential of the NOP receptor as a therapeutic target. Ro 64-6198 is systemically active and achieves high brain penetration. It has subnanomolar affinity for the NOP receptor and is at least 100 times more selective for the NOP receptor over the classic opioid receptors. Ro 64-6198 ranges from partial to full agonist, depending on the assay. Preclinical data indicate that Ro 64-6198 may have broad clinical uses, such as in treating stress and anxiety, addiction, neuropathic pain, cough, and anorexia. This review summarizes the pharmacology and preclinical data of Ro 64-6198. PMID- 17461894 TI - Coming together to conquer the fight against breast cancer in countries of limited resources: the challenges and the opportunities. PMID- 17461895 TI - Maximizing breast projection with combined free nipple graft reduction mammaplasty and back-folded dermaglandular inferior pedicle. AB - Standard technique for free nipple reduction mammoplasty was described by Thorek in 1922. In contrast to its effectiveness, late postoperative results included insufficient projection of the breast and the nipple-areola region. We describe a modification of this well recognized technique in order to increase central mound projection and improve nipple-areola projection by suturing the dermaglandular flap to the pectoralis major muscle by back-folding the pedicle. Twenty macromastia patients were subjected to free-nipple-graft reduction mammoplasty in combination with inferior pedicled dermaglandular reduction mammaplasty of a total of 40 breasts with this technique between years 2000 and 2004. Preoperative planning for inferior pedicled dermaglandular flap was made using the "Wise" pattern for large breasts. The variation of the technique comes from using the back-folded deepithelialized inferior pedicled dermaglandular flap for increasing the breast mound projection by fixating the demaglandular flap with absorbable sutures to the underlying pectoralis major muscle fascia and the costal cartilage pericondrium. By applying this technique, increased projection during the early preoperative and late postoperative periods are achieved, compared with patients who only underwent free-nipple- graft reduction mammoplasty. PMID- 17461896 TI - Factors determining adequacy of axillary node dissection in breast cancer patients. AB - With increased focus on quality assurance, a complete axillary lymph node dissection (ALND) has been defined as the removal of 10 or more lymph nodes (LN). The objective of this study was to determine which patient, physician, and geographic factors predict the adequacy of ALND in breast cancer patients. The University of Louisville Breast Cancer Sentinel Lymph Node Study is a multicenter, prospective study of 4,131 patients, all of whom had a sentinel node biopsy and completion ALND. Univariate and multivariate analyses were performed to determine which factors were independently associated with the removal of 10 or more LN. Of the 4,131 patients in this study, the median number of LN removed was 11 (range; 3-45). Ten or more LN were removed in 3,213 (77.8%) patients. The median patient age in this study was 60 (range; 27-100), with a median tumor size of 1.5 cm (range; 0.1-11.0 cm). On univariate analysis, patient age, tumor size, and palpability were correlated with adequacy of ALND. Academic affiliation and percentage of breast practice were significant physician factors predictive of adequacy of ALND. Both geographic region and community size were significantly correlated with adequacy of ALND. On multivariate analysis, patient age (p = 0.024), surgeon academic affiliation (p < 0.001), percentage breast practice (p < 0.001), and community size (p = 0.003) were significant determinants of adequacy of ALND. Younger patients were more likely to have an adequate ALND. Surgeons in academic practice had a higher rate of adequate ALND, as did those practicing in larger communities. Surgeons with a more breast experience had a lower rate of adequate ALND. Patient age, surgeon academic affiliation, and breast experience, as well as community size are all significant factors predictive of adequacy of ALND. PMID- 17461897 TI - Body image score following anterior and lateral approaches to wide local excision for early breast cancer. AB - Breast cancer surgery is an emotive topic and rather than just simple cosmetic issues, the final appearance of the operated breast has been shown to have a significant impact on psychologic well-being and overall quality of life. Wide local excision (WLE) is a popular conservative approach to small tumors, which do not involve the nipple-areolar complex. This study compares WLE via an anterior and lateral approach. A retrospective survey of patient satisfaction was performed using a recognized and validated Body Image Score (BIS), which was sent to a total of 267 patients who had undergone WLE by either the anterior or lateral approach. The response rate was 57%, of which 89 women had undergone surgery via the anterior and 80 by the lateral approach. There was no significant difference in BIS when comparing the anterior and lateral approaches, despite concerns regarding the physical appearance of the scar and postoperative breast expressed by patients. Generally, surgery on the upper part of the breast was associated with less satisfaction than the lower, but "hiding" the scar with a lateral approach did not improve satisfaction. Surgeons should be reassured that the approach to WLE of early breast cancers has no impact on the patients overall body image and that the choice of technique should be based on the patient's personal preference and the surgeons experience and skill. PMID- 17461898 TI - Neoadjuvant use of endocrine therapy in breast cancer. AB - Neoadjuvant endocrine therapy is becoming increasingly popular as a safe and effective alternative to chemotherapy in selected patients. Large randomized studies have been published comparing tamoxifen with steroidal and nonsteroidal aromatase inhibitors, with favorable results for aromatase inhibitors letrozole, anastrozole, and exemestane. Endocrine therapy can be used in the neoadjuvant setting for conversion of inoperable breast tumors to operable, and from potential mastectomy to breast-conserving surgery. The use of endocrine agents in this setting also provides an opportunity for the study of their biological effects upon tumor. PMID- 17461899 TI - Predictors of residual invasive disease after core needle biopsy diagnosis of ductal carcinoma in situ. AB - Core needle biopsy (CNB) is used to sample both mammographically and ultrasound detected breast lesions. A diagnosis of ductal carcinoma in situ (DCIS) by CNB does not ensure the absence of invasive cancer upon surgical excision and as a result an upstaged patient may need to undergo additional surgery for axillary nodal evaluation. This study evaluates the accuracy of CNB in excluding invasive disease and the preoperative features that predict upstaging of DCIS to invasive breast cancer. Two hundred fifty-four patients over an 8-year period from 1994 to 2002 with a diagnosis of DCIS alone by CNB were retrospectively reviewed. Underestimation of invasive cancer by CNB was determined. Radiographic, pathologic, and surgical features of the cohort were compared using univariate and multivariate analysis. The mean age was 55 years (range 27-84) and mean follow-up was 25 months with one patient unavailable for follow-up. There were a total of six patient deaths, all of which were not disease-specific. A total of 21 out of 254 patients (8%) with DCIS by CNB were upstaged to invasive cancer following surgical excision. There was a significant inverse relationship between the number of core biopsies and the incidence of upstaging (p < 0.006) in that patients with fewer core samples were more likely to be upstaged at surgical pathology. No relationship was noted between the size of the core samples and the likelihood of upstaging (p > 0.4). Of 21 patients with invasion, all but two had comedonecrosis by CNB. Comedonecrosis by CNB significantly increased the likelihood of upstaging (p < 0.001). Of the 21 patients who were upstaged, 12 required subsequent surgery for nodal evaluation while nine had sentinel node biopsy at initial operation. Finally, upstaged patients were significantly more likely to have a positive margin (p < 0.008). Ductal carcinoma in situ with comedonecrosis on CNB can help to predict the possibility of invasion. Increasing the number of core biopsies reduced the likelihood of sampling error. PMID- 17461900 TI - Longitudinal patterns of weight gain after breast cancer diagnosis: observations beyond the first year. AB - Many, but not all patients experience weight gain 1 year after a breast cancer diagnosis; clearly defined, clinically relevant groups at risk of weight gain have yet to be described. We set out to determine the factors associated with weight gain over time in patients with invasive breast cancer during a period of predominantly anthracycline-based adjuvant chemotherapy and to identify groups with differing weight gain risks. Breast cancer patients (stage I-IIIB) were identified in a retrospective chart review. Evaluated parameters included weight at diagnosis and 1, 2, and 3 years later, height, body mass index (BMI), age, menopausal and change in menopausal status, as well as therapy and pathologic stage. Regression models identified significant independent predictors of weight change. Recursive partitioning analysis (RPA) was employed to divide the dataset into relevant and significant groups. In 185 identified patients, regression models and RPA demonstrated that weight gain at 1 year was associated with younger age, adjuvant chemotherapy, and lower BMI. Weight gain at 2 years (n = 176) was greater than at year 1, and in addition to weight gain at year 1, was associated with younger age and adjuvant chemotherapy in regression analysis; RPA found that anthracycline therapy, age, and BMI were important. Weights at 3 years were similar to those seen at 2 years. Early-stage breast cancer patients treated with chemotherapy continue to gain weight 2 years after diagnosis, and this weight gain appears to be persistent at year 3. Observation beyond 1 year is needed to adequately evaluate weight gain in early-stage breast cancer patients, particularly for those receiving contemporary adjuvant chemotherapy. PMID- 17461901 TI - Breast edema in patients undergoing breast-conserving treatment for breast cancer: assessment via high frequency ultrasound. AB - To identify factors that can influence breast edema in women undergoing breast conserving therapy. Breast edema was assessed clinically and via high frequency ultrasound (HFUS) prior to, during and following radiotherapy. Fifty-four women were assessed. Breast edema was present prior to radiotherapy in patients who had undergone level 2 node dissection or had wound infection after sentinel node dissection. Edema increased during and after radiotherapy and peaked at 4-6 months. The time course of breast edema was related to the extent of nodal dissection, postoperative wound infection and regional radiotherapy. HFUS prior to irradiation was found to be no better than clinical assessment in predicting prolonged parenchymal breast edema but was significantly better at the end of irradiation. Breast edema levels are minimal in patients who do not undergo axillary node dissection or have an uncomplicated sentinel node dissection. Most edema is due to compromise of the draining lymphatics, which relates largely to the extent of axillary node dissection. HFUS appears to be a useful in the research setting in quantifying the effect of techniques that aim to reduce complications such as edema. PMID- 17461902 TI - Phase I/II trial of adjuvant dose-dense docetaxel/epirubicin/cyclophosphamide (TEC) in stage II and III breast cancer. AB - This phase I/II trial investigates the safety and feasibility of six cycles of concurrent taxane, anthracycline and cyclophosphamide on a dose dense schedule. Patients with stage II/III breast cancer were treated with docetaxel (T) 75 mg/m(2), epirubicin (E) 75 mg/m(2) (cohort 1, n = 3) or 100 mg/m(2) (cohort 2, n = 12), and cyclophosphamide (C) 500 mg/m(2) IV on day 1, with pegfilgrastim 6 mg subcutaneously on day 2, every 2 weeks for six cycles. Patients were assessed for toxicity every 2 weeks; cardiac function and response (if neoadjuvant) were assessed after six cycles. All patients in cohort 1 received 100% planned dose intensity; in cohort 2, five of twelve patients received 100% and 11/12 received >80%. There were no dose reductions or delays for day 1 myelotoxicity. Dose reductions as a result of febrile neutropenia (FN) occurred in cohort 2, with six of twelve patients experiencing FN in seven of sixty-nine cycles. Six patients had anemia > or =grade 3; five received RBC transfusion and seven received an erythropoietic growth factor. Four patients required dose reductions for nonhematologic toxicity (two mucositis; one neurotoxicity; one diarrhea + cellulitis). Four patients developed thrombophlebitis, which was associated with FN in one of four. Two of fourteen evaluable patients had asymptomatic decreases in LVEF >10%; all remained within normal range. All four patients receiving neoadjuvant TEC had significant clinical responses (one CR, three PR). No pathologic CRs were seen. CONCLUSIONS: Dose dense TEC chemotherapy is feasible, has acceptable toxicity at doses equivalent to TAC (docetaxel 75 mg/m(2), epirubicin 75 mg/m(2), cyclophosphamide 600 mg/m(2)), and has moderate but manageable toxicity using a higher epirubicin dose of 100 mg/m(2), with FN occurring in six of twelve patients at the higher dose. PMID- 17461903 TI - Ductal pattern enhancement on magnetic resonance imaging of the breast due to ductal lavage. AB - Our purpose is to describe the appearance of breast ductal enhancement found on magnetic resonance imaging (MRI) after breast ductal lavage (DL). We describe a novel etiology of enhancement in a ductal pattern on postcontrast MRI of the breast. Knowledge of the potential for breast MRI enhancement subsequent to DL, which can mimic the appearance of a pathologic lesion, is critical to the care of patients who undergo breast MRI and DL or other intraductal cannulation procedures. PMID- 17461904 TI - Safety of elective hand surgery following axillary lymph node dissection for breast cancer. AB - The development of lymphedema is the most feared complication shared by breast cancer survivors undergoing hand surgery after prior axillary lymph node dissection (ALND). Traditionally, these patients are advised to avoid any interventional procedures in the ipsilateral upper extremity. However, the appropriateness of some of these precautions was recently challenged by some surgeons claiming that elective hand operations can be safely performed in these patients. The purpose of this study was to evaluate our experience and determine the safety of elective hand operations in breast cancer survivors. The medical records of patients operated for different hand conditions after prior breast surgery and ALND at our institution between 1983 and 2002 were reviewed. The techniques and preventive measures performed, use of antibiotics, and upper extremity complications associated with the operations were analyzed. Overall, we operated on 27 patients after prior ALND performed for breast cancer. Follow-up was available for 25 patients. Four patients had pre-existing lymphedema. The surgical technique used was similar to that performed in patients without prior ALND and antibiotic prophylaxis was not given. Delayed wound healing was observed in one patient and finger joint stiffness in another. Two patients with pre existing lymphedema developed temporary worsening of their condition. None of the patients developed new lymphedema. The results of the present study support the few previous studies, suggesting that hand surgery can be safely performed in patients with prior ALND. Based on these findings, the appropriateness of the rigorous precautions and prohibitions regarding the care and use of the ipsilateral upper extremity may need to be reconsidered. PMID- 17461905 TI - Extramammary Paget's disease of the axilla: an unusual case. AB - Extramammary Paget's disease is a rare lesion, often involving the skin of the genital or perianal regions. Less commonly, it has been reported to affect the skin of the axilla. There are very few other cases of extramammary Paget's disease reported in the literature, and the appropriate use of newer techniques such as magnetic resonance imaging and sentinel lymph node biopsy in this setting is not well-studied. We present a case of extramammary Paget's disease of the axilla, and discuss the known literature regarding this rare disease. PMID- 17461906 TI - An unusual presentation of locally recurrent breast cancer: a case report. AB - Recurrent breast carcinoma is usually detected by imaging studies and biopsy. We present a case with unusual clinical presentation and discuss the diagnosis and treatment. While core needle biopsy and fine-needle aspiration are important in the diagnosis of early-stage breast cancer, physical examination and close follow up are important in the absence of a diagnosis. PMID- 17461907 TI - Chrysomya bezziana: a rare infestation of the breast. PMID- 17461908 TI - Morphea of the breast: case reports and review of literature. AB - Morphea of the breast is an uncommon skin condition associated with autoimmune disorders and has also been described following radiotherapy. The inflammatory stage of morphea can commonly be mistaken for a breast abscess or inflammation. We report two cases of localized scleroderma affecting the breast with no history of autoimmune disease or radiation treatment. In both the cases, the lesions improved with the use of topical steroids. PMID- 17461909 TI - Lymph node axillary metastasis from occult contralateral infiltrating lobular carcinoma arising in accessory breast: MRI diagnosis. AB - We describe a case of right axillary lymph node metastasis of an occult infiltrating lobular carcinoma arising from accessory mammary gland of the left upper anterior chest wall. Ultrasonography and mammography were normal bilaterally. Magnetic resonance imaging (MRI) revealed a 3.34 cm inhomogeneous lesion. Then, core biopsy under ultrasound guidance demonstrated a typical infiltrating breast lobular carcinoma. To our knowledge, this is the first case reported in the literature of an axillary lymph node metastasis from an occult contralateral infiltrating lobular carcinoma of the accessory breast tissue. MRI was useful for assessing the lesion of the accessory breast tissue. PMID- 17461910 TI - Megakaryocytes in sentinel lymph node--a potential source for diagnostic error. PMID- 17461911 TI - Multiple myeloma recurrence presenting as bilateral breast masses. PMID- 17461912 TI - Malignant phyllodes tumor of a genotypic male, phenotypic female with liposarcomatous differentiation. PMID- 17461913 TI - Synchronous benign and malignant mesenchymal breast tumor. PMID- 17461914 TI - Renal cell carcinoma metastasis to the breast: mammographic, sonographic, CT, and pathologic correlation. PMID- 17461915 TI - The Fakous experience: meeting the challenge of cancer in a low resource setting. PMID- 17461916 TI - Studying the accuracy of breast physical examination and breast physical examination along with mammography in the early detection of breast masses among Iranian women. PMID- 17461917 TI - An unusual case of breast liposarcoma with liver metastases: the role of radical surgery. PMID- 17461918 TI - Paradigms in oncoplastic breast surgery: a careful assessment of the oncological need and esthetic objective. PMID- 17461919 TI - Post-traumatic stress disorder and comorbid depression among survivors of the 1999 earthquake in Turkey. AB - This study examined the prevalence of post-traumatic stress disorder (PTSD) and comorbid depression some three years after the August 1999 earthquake in Turkey among a sample of 769 survivors relocated to a permanent housing site built for homeless survivors in the epicentre region. Time since trauma was 3.1 years for 81 per cent of the participants and 3.9 years for the remainder. Survivors were assessed using the Screening Instrument for Traumatic Stress in Earthquake Survivors, an easily administered self-rating scale with demonstrated validity. The estimated rates of PTSD and comorbid depression were 40 per cent and 18 per cent, respectively. Linear regression analyses showed that PTSD strongly related to fear during the earthquake, while depression related to loss of family members. These results suggest that catastrophic earthquakes have long-term psychological consequences and highlight the need for a cost-effective mental health care model for earthquake survivors. PMID- 17461920 TI - Characteristics, context and risk: NGO insecurity in conflict zones. AB - This paper reports on research conducted on the insecurity of non-governmental organisations (NGOs) between 1999 and 2002, with the goal of contributing to the debate on the reasons why NGO actors are targets of violence in conflict settings. The research involved the collection of data from three countries Angola, Ecuador and Sierra Leone-and exploration of the relationship between levels of insecurity, context and the characteristics of NGOs. Four risk factors appear to heighten the degree of insecurity that NGOs face: (1) carrying out multiple types of activities and providing material aid; (2)'operationality'(that is, implementing programmes and activities); (3) working with both sides of the conflict; and (4) integrating into the local community. The paper discusses the methodological approach adopted for the research, the differences between ambient and situational insecurity and the findings related to risk factors. It concludes with a summary of the study's implications. PMID- 17461921 TI - Gender and Hurricane Mitch: reconstructing subjectivities after disaster. AB - Much of the gender and disaster literature calls for more gender-sensitive disaster relief and research by focusing on the ways in which women are more vulnerable in a disaster or on their unique capabilities as community leaders or natural resource managers, which are often overlooked or underutilised in emergency management strategies. As well as seeking to overcome the (strategic) essentialism that is part of these calls and debates, this paper pays closer attention to gender identity and subjectivity as these are constructed and reworked through the disaster process to highlight the complexities and contradictions associated with women's responses to a disaster. This focus, while crucial to gaining a deeper understanding of the gendered dimensions of disaster, also complicates attempts to create more gender-sensitive frameworks for disaster response. It draws on qualitative research conducted with a number of women in the wake of Hurricane Mitch (1998) in Nicaragua. PMID- 17461922 TI - A tool to aid emergency managers and communities in appraising private dam safety and policy. AB - Issues concerning dam safety and equitable sharing of catchment run-off are receiving more attention throughout the world. This paper assesses these matters in the context of Australia, and the need for policy responses. Landholders often overlook the common law obligation to review/design dams to current standards because of high costs, leaving them vulnerable to litigation if their dam fails. The paper reports on an innovative spillway design/review procedure, applicable to southeast Australia, but transferable to any region worldwide. Dam safety policy models and guidelines derived from international best practice are linked to the procedure and intended to aid government decision-making. The procedure minimises costs to landholders and provides an acceptable level of safety assurance to downstream communities. Also discussed are recent surveys testing community attitudes to the procedure and implemented dam safety and water allocation policies. These further guide any government wanting to implement this'integrated engineering and community partnerships'approach to preventing potential disasters due to private dam failure and achieving sustainable and safe water storage and use. PMID- 17461923 TI - The Prestige oil spill and its economic impact on the Galician fishing sector. AB - The sinking of the Prestige oil tanker on 18 November 2002 off the coast of Galicia, Spain, had important economic, environmental and social ramifications. The aim of this paper is to carry out an initial analysis of the costs related to a halt in fishing activities in Galicia between November 2002 and December 2003. This involves three different steps: an assessment of the cost of the preventative and palliative measures introduced by Spanish public administrations (compensation for affected fishermen and shellfish fisherman); an indirect evaluation of the implications of the disaster (via a study of data on production); and a direct appraisal of the economic impact of the event (reduction in income), using questionnaires completed by a representative sample of fishermen and shellfish fisherman. The results obtained from these three methods of estimating losses are compatible. By December 2003, losses to the Galician fishing sector stood at an estimated EUR 76 million. PMID- 17461924 TI - Treatment of acquired hemophilia A. AB - Acquired hemophilia A (AH) is an autoimmune disease that leads to potentially severe bleeding. Management relies on rapid and accurate diagnosis, control of bleeding episodes and eradication of the inhibitor by immunosuppression. There is extensive literature about the disease but only few controlled data are available. This paper reviews the current literature on treatment strategies for hemostatic therapy and inhibitor eradication. Potential future developments are discussed. PMID- 17461925 TI - Does ATP act through P2X(1) receptors to regulate platelet activation and thrombus formation? PMID- 17461926 TI - Survey of lupus anticoagulant diagnosis by central evaluation of positive plasma samples. AB - OBJECTIVE: To determine whether the diagnosis of lupus anticoagulant (LAC) in a large cohort of positive patients was confirmed at a reference laboratory. METHODS: Over a 1-year period, each participating center collected samples from LAC-positive patients. Plasma was filtered and kept deep-frozen until it was sent on dry ice to the reference laboratory by express courier. Centers returned detailed laboratory information and clinical data from each patient. The reference laboratory screened plasma samples by diluted Russell viper venom time (dRVVT) and kaolin clotting time (KCT). When these were prolonged, 1:1 mixing studies were carried out, and confirmatory tests were performed as appropriate. Positive samples were further tested by thrombin time (TT). The presence of heparin was checked by measuring antifactor Xa activity when TT was prolonged. Negative samples were tested by activated partial thromboplastin time using hexagonal phospholipids. RESULTS: Plasma samples from 302 patients from 29 anticoagulation clinics were analyzed. LAC was excluded in 71 samples (24%), because dRVVT and KCT screening test results were normal (34) or reversed to normal by mixing studies (35). The remaining two samples were considered negative because they contained heparin. LAC-negative patients showed different characteristics from those in whom diagnosis was confirmed. They were significantly older (49.7 vs. 45.0 years, P < 0.03), were more often first diagnosed (66% vs. 41%, P < 0.001), and were more frequently judged as mild in LAC potency (60% vs. 25%, P < 0.0001). Moreover, anticardiolipin and anti-beta(2) glycoprotein I antibody values were more often normal in LAC-negative (82%) than in LAC-positive (42%) samples (P < 0.0001). LAC-positive samples identified by both dRVVT and KCT (146/231, 63%) showed a LAC potency that was significantly stronger than that in samples in which LAC diagnosis was made by a single test. CONCLUSIONS: A false-positive LAC diagnosis is not uncommon across specialized centers. Patients' characteristics and a complete antiphospholipid antibody profile may help to identify these individuals. PMID- 17461927 TI - Frequency of renal impairment, advanced age, obesity and cancer in venous thromboembolism patients in clinical practice. AB - BACKGROUND: Low-molecular-weight heparin (LMWH) dosed by weight is recommended as first-line therapy for the initial treatment of venous thromboembolism (VTE) and as monotherapy for long-term treatment of cancer-related VTE. In 'special populations' such as those with renal impairment or the elderly, weight-based dosing may be excessive, and capping the dose in obese patients may lead to inadequate dosing. OBJECTIVES: We determined the frequency of 'special population' characteristics (renal impairment, advanced age, obesity) and cancer among VTE patients in clinical practice, and assessed whether these characteristics appeared to influence the type and dose of anticoagulants prescribed. METHODS: During 2004-2005, among consecutive patients with VTE at two large Canadian hospitals, the proportions with the above characteristics were calculated and treatments prescribed were determined. RESULTS: Of 524 VTE patients, 31% were aged > 75 years. Moderate renal impairment [creatinine clearance (CrCl) 30-59 mL min(-1)] was present in 20% of patients, and severe renal impairment (CrCl < 30 mL min(-1)) in 5% of patients. LMWH was prescribed to 67% of patients with severe renal impairment and to 83% of patients with moderate renal impairment. Body weight was > 100 kg in 15% of patients. Underdosing of LMWH by > 10% was documented in 36% of such patients compared with 8% of patients < 100 kg (P < 0.001). Among 26% of patients with active cancer, only one-third were prescribed LMWH monotherapy. CONCLUSIONS: In clinical practice, renal impairment, advanced age, obesity and cancer are frequently present in patients with VTE. A considerable proportion of these patients may not receive the optimal type or dose of medication to treat VTE. PMID- 17461928 TI - Effects of pioglitazone on fasting and postprandial levels of lipid and hemostatic variables in overweight non-diabetic patients with coronary artery disease. AB - OBJECTIVES: To evaluate the effects of pioglitazone on insulin sensitivity and levels of biomarkers associated with thrombotic risk in overweight and obese, non diabetic subjects with coronary artery disease. BACKGROUND: Little information is available regarding the effects of thiazolidinediones in the absence of diabetes. Further, although postprandial hyperlipemia is a risk factor for cardiovascular diseases, there is limited information about the postprandial effects. METHODS: Twenty overweight and obese, non-diabetic patients with coronary artery disease were enrolled in a randomized, placebo-controlled, double-blind study. Subjects were on atorvastatin for the duration of the study and received either placebo or pioglitazone (45 mg day(-1)) for 12 weeks and then crossed over to the alternative therapy for an additional 12 weeks. Insulin sensitivity, fasting and postprandial levels of lipid, hemostatic, and inflammatory variables were measured, and endothelial function was assessed. RESULTS: Insulin sensitivity improved from 0.03 micromol kg(-1) x min pM(-1) on placebo to 0.04 on pioglitazone (P = 0.0002), and there were decreases in fasting levels of factor (F) VII:C (102 +/- 17% to 92 +/- 18%, P = 0.001), FVII:Ag (68 +/- 12% to 60 +/- 14%, P = 0.01) and in von Willebrand factor (VWF) (174 +/- 94% to 142 +/- 69%, P = 0.01). Pioglitazone lowered postprandial levels of FVII:Ag, FVII:C, plasminogen activator inhibitor-1, VWF, and triglycerides, and increased high-density lipoproteins (+9%, P = 0.02). CONCLUSIONS: Pioglitazone improves insulin sensitivity and favorably modifies fasting and postprandial lipid, hemostatic and inflammatory markers of the metabolic syndrome in overweight and obese non diabetic patients with coronary artery disease. PMID- 17461929 TI - Effect of the anti-factor Xa and anti-factor IIa activities of low-molecular weight heparins upon the phases of thrombin generation. AB - BACKGROUND: Low-molecular-weight heparins (LMWHs), derived from unfractionated heparin (UFH) by different depolymerization procedures, vary in both their relative abilities to enhance the inhibition of FXa (anti-FXa) and thrombin (anti FIIa), and in their physicochemical properties. OBJECTIVE: We aimed to profile the inhibition of thrombin generation induced by bemiparin, enoxaparin, nadroparin, dalteparin and tinzaparin in platelet-rich plasma (PRP), and to compare them with UFH and fondaparinux (a synthetic pentasaccharide that specifically enhances FXa inhibition). METHODS: Different LMWHs, UFH or fondaparinux were added to normal PRP. Thereafter, tissue factor-triggered thrombin generation was assessed using the Thrombogram-Thrombinoscope assay. RESULTS: At equivalent anti-FIIa activity concentrations, LMWHs and UFH exhibited similar inhibitory effects upon thrombin generation. However, when used at equivalent anti-FXa activity concentrations, tinzaparin was significantly more active than the other LMWHs at inhibiting thrombin generation, and had similar activity to that of UFH. Enoxaparin, nadroparin and dalteparin all showed similar inhibitory activities. In these experiments, bemiparin exhibited the lowest inhibitory effect on thrombin generation of all the LMWHs. At 0.1 microg mL(-1) (0.093 anti-FXa IU mL(-1)), fondaparinux inhibited the rate of thrombin generation by 50%. A 7-fold higher concentration of fondaparinux was required to inhibit the endogenous thrombin potential by 50%. CONCLUSIONS: LMWHs have a variable inhibitory effect on thrombin generation in vitro when compared by anti FXa activity, but are similar when compared by their anti-FIIa activities. The rate of thrombin generation during the propagation phase, rather than the endogenous thrombin potential, is more sensitive to the anticoagulant activity of fondaparinux and the polysaccharide chains of LMWHs possessing only anti-FXa activity. PMID- 17461930 TI - Differential effects of high prothrombin levels on thrombin generation depending on the cause of the hyperprothrombinemia. AB - BACKGROUND: Hyperprothrombinemia, resulting from the prothrombin G20210A mutation or other causes, is associated with activated protein C (APC) resistance and increased thrombosis risk. When high prothrombin levels are a result of increased hepatic biosynthesis, these effects may be counteracted by concomitantly increased levels of the anticoagulant factors (particularly protein S). Differently, in prothrombin G20210A carriers only prothrombin levels are elevated. OBJECTIVE: To investigate whether prothrombin G20210A carriers have a more severe hypercoagulable state than non-carriers with comparable prothrombin levels. PATIENTS/METHODS: Coagulation factor levels, thrombin generation (Calibrated Automated Thrombogram in the presence and absence of APC) and APC resistance were measured in normal (n = 132), heterozygous (n = 167) and homozygous (n = 3) individuals. RESULTS: Prothrombin levels, thrombin generation and APC resistance were higher in carriers of the prothrombin G20210A mutation (especially those who had experienced venous thrombosis) than in non-carriers, whereas protein S and antithrombin levels were similar among genotype groups. Because individuals with high prothrombin levels in the absence of the prothrombin G20210A mutation tend to have all liver-synthesized factors elevated, carriers of the mutation had lower protein S and antithrombin levels than non carriers with equally high prothrombin levels. Accordingly, they also generated more thrombin and showed a tendency toward higher APC resistance. Analogous effects, but less pronounced, were observed in homozygotes for the prothrombin A19911G polymorphism, which also upregulates prothrombin levels. CONCLUSIONS: Individuals with hyperprothrombinemia as a result of prothrombin gene mutations generate more thrombin and tend to be more APC-resistant than individuals with comparable prothrombin levels because of other causes. PMID- 17461931 TI - The effect of fibrinogen concentrate on thrombocytopenia. AB - OBJECTIVES: The hypothesis that the administration of fibrinogen concentrate enables restoration of impaired clot formation and increased bleeding in severe thrombocytopenia was tested. METHODS: Thirty pigs were anesthetized, instrumented for blood sampling (routine coagulation tests, modified thrombelastography ROTEM, hemodynamic monitoring and platelet apheresis to a target below 30 x 10(9) L(-1) after splenectomy. Thereafter 10 each of the animals randomly received two apheresis platelet concentrates, 250 mg kg(-1) fibrinogen concentrate or normal saline solution. A standardized liver injury was subsequently inflicted to induce uncontrolled hemorrhage. RESULTS: Median (Q1, Q3) clot firmness increased significantly more in thrombocytopenic pigs after fibrinogen administration (42 mm (41, 43) to 60 mm (57, 63)) than following platelet transfusion (40 mm (37, 45) to 52 mm (48, 55), P = 0.0004) or placebo (45 mm (41, 48) to 45 mm (43, 46), P = 0.0002). Median blood loss velocity after liver injury was significantly less with fibrinogen (33 mL min(-1), P = 0.005) than with platelets (62 mL min(-1), P = 0.037) or saline (84 mL min(-1), P = 0.005), and median survival time after liver injury was 55 min in the fibrinogen, 26 min in the platelet (P = 0.035) and 19 min in the saline group (P = < 0.0001). CONCLUSIONS: These data show for the first time that impaired clot formation during thrombocytopenia improves with administration of fibrinogen concentrate, which results in a slowdown of blood loss and prolonged survival. PMID- 17461932 TI - A role for glycosphingolipid-enriched microdomains in platelet glycoprotein Ib mediated platelet activation. AB - BACKGROUND: Glycoprotein (GP) Ib, a platelet von Willebrand factor (VWF) receptor, plays a crucial role in thrombosis and hemostasis. As recent reports have suggested that GPIb partially locates in a particular region, designated as glycosphingolipid-enriched microdomains (GEMs), we hypothesized that GEMs play a central role in GPIb-mediated platelet activation. METHODS: Platelets were stimulated by VWF/botrocetin to activate platelets through GPIb. GEMs and non GEMs were isolated by sucrose density gradient ultracentrifugation and the location of signaling molecules characterized. The role of GEMs-mediated signaling in platelet behavior was tested by platelet aggregation and by platelet interaction with immobilized VWF under flow conditions when GEMs were disrupted by methyl-beta-cyclodextrin (MbetaCD). RESULTS: GPIb was partially translocated to GEMs upon VWF/botrocetin stimulation. Immunoprecipitation of GPIb in GEMs and non-GEMs revealed that the tyrosine kinases, Src and Lyn, were associated with GPIb only in GEMs after GPIb-stimulation, and not in non-GEMs. Activation of PLCgamma2 was more intense in GEMs than non-GEMs. Disruption of GEMs by MbetaCD strongly inhibited tyrosine phosphorylation of Syk and PLCgamma2. Functional studies revealed that stable adhesion of platelets to a VWF-coated surface under flow was impaired by GEM disruption by MbetaCD. CONCLUSION: The combined results suggest that GEMs play an important role in GPIb-mediated platelet activation. PMID- 17461933 TI - Effects of plasma membrane Ca(2+) -ATPase tyrosine phosphorylation on human platelet function. AB - BACKGROUND: The plasma membrane Ca(2+)-ATPase (PMCA) plays an essential role in maintaining low intracellular Ca(2+) ([Ca(2+)](i)) in resting platelets. Earlier studies demonstrated that platelet activation by thrombin results in tyrosine phosphorylation of PMCA, which inhibits pump activity. OBJECTIVES: The objective was to determine the functional consequences of PMCA tyrosine phosphorylation. METHODS: A decapeptide including the tyrosine phosphorylation site of PMCA and a scrambled version were synthesized and introduced into human platelets using saponin. Fura-2 calcium monitoring and aggregometry were used to characterize the effects of inhibition of tyrosine phosphorylation. RESULTS: Western blot analysis of immunoprecipitates showed that introduction of the inhibitory peptide decreased tyrosine phosphorylation of PMCA by nearly 60% in saponin permeabilized, thrombin-treated platelets as compared with the scrambled control peptide. Concomitant with inhibition of PMCA tyrosine phosphorylation was a significant decrease in [Ca(2+)](i) during thrombin-mediated platelet activation. The functional consequence of reduced PMCA tyrosine phosphorylation and decreased [Ca(2+)](i) was a significant delay in the onset of thrombin-mediated platelet aggregation. CONCLUSIONS: The results demonstrate that PMCA tyrosine phosphorylation regulates [Ca(2+)](i) during platelet activation, which affects downstream events in the activation process. Moreover, PMCA tyrosine phosphorylation and resultant inhibition of PMCA activity produces a positive feedback loop mechanism by enhancing the increase in [Ca(2+)](i) accompanying platelet activation. PMID- 17461934 TI - Thrombin enhances herpes simplex virus infection of cells involving protease activated receptor 1. AB - BACKGROUND: We have previously shown that the surface of purified herpes family viruses can initiate thrombin production by expressing host-encoded and virus encoded procoagulant factors. These enable the virus to bypass the normal cell regulated mechanisms for initiating coagulation, and provide a link between infection and vascular disease. OBJECTIVE: In the current study we investigated why these viruses may have evolved to generate thrombin. METHODS: Using cytolytic viral plaque assays, the current study examines the effect of thrombin on human umbilical vein endothelial cell (HUVEC) or human foreskin fibroblast (HFF) infection by purified herpes simplex virus type 1 (HSV1) and type 2 (HSV2). RESULTS: Demonstrating that the availability of thrombin is an advantage to the virus, purified thrombin added to serum-free inoculation media resulted in up to a 3-fold enhancement of infection depending on the virus strain and cell type. The effect of thrombin on HUVEC infection was generally greater than its effect on HFF. To illustrate the involvement of thrombin produced during inoculation, hirudin was shown to inhibit the infection of each HSV strain, but only when serum containing clotting factors for thrombin production was present in media. The involvement of protease-activated receptor 1 (PAR1) was supported using PAR1 activating peptides in place of thrombin and PAR1-specific antibodies to inhibit the effects of thrombin. CONCLUSION: These data show that HSV1 and HSV2 initiate thrombin production to increase the susceptibility of cells to infection through a mechanism involving PAR1-mediated cell modulation. PMID- 17461935 TI - Atypical mutations of the prothrombin gene at positions 20,209 and 20,218, and a novel mutation at position 20,219. Report on 10 patients. PMID- 17461936 TI - A case of profound and prolonged tirofiban-induced thrombocytopenia and its correction by intravenous immunoglobulin G. PMID- 17461937 TI - Folic acid and B vitamins improve hyperhomocysteinemia-induced cardiovascular risk profile in renal transplant recipients. PMID- 17461938 TI - Alternative diagnosis other than pulmonary embolism as a subjective variable in the Wells clinical decision rule: not so bad after all. PMID- 17461939 TI - Simple coagulation tests improve early mortality prediction for patients in intensive care units who have proven or suspected septic shock. PMID- 17461940 TI - Evidence that pre-existent variability in platelet response to ADP accounts for 'clopidogrel resistance': a rebuttal. PMID- 17461941 TI - Outcomes-based patient care in veterinary surgery: what is an outcome measure? PMID- 17461942 TI - Beating heart mitral valve replacement with a bovine pericardial bioprosthesis for treatment of mitral valve dysplasia in a Bull Terrier. AB - OBJECTIVES: To describe an open, beating heart surgical technique and use of a bovine pericardial prosthetic valve for mitral valve replacement (MVR) in the dog. STUDY DESIGN: Clinical case report. ANIMALS: Male Bull Terrier (17-month old, 26 kg) with mitral valve dysplasia and severe regurgitation. METHODS: A bovine pericardial bioprosthesis was used to replace the mitral valve using an open beating heart surgical technique and cardiopulmonary bypass. RESULTS: Successful MVR was achieved using a beating heart technique. Mitral regurgitation resolved and cardiac performances improved (left ventricular end-diastolic diameter decreased from 57.6 to 48.7 mm, and left atrium/aorta ratio returned to almost normal, from 1.62 to 1.19). Cardiopulmonary by-pass time and total surgical duration were decreased compared with standard cardioplegic techniques. Surgical recovery was uneventful and on echocardiography 6 months later valve function was excellent. CONCLUSION: Considering the technique advantages (no cardiac arrest, ischemic reperfusion injury, and hypothermia, or the need for aortic dissection and cannulation for administration of cardioplegic solution), short-term mortality and morbidity may be reduced compared with standard cardioplegic techniques. CLINICAL RELEVANCE: Based on experience in this dog, beating heart mitral valvular replacement is a seemingly safe and viable option for the dog and bovine pericardial prosthesis may provide better long-term survival than mechanical prostheses. PMID- 17461943 TI - Surgical and interventional radiographic treatment of dogs with hepatic arteriovenous fistulae. AB - OBJECTIVE: To report outcome after surgical and interventional radiographic treatment of hepatic arteriovenous fistulae (HAVF) in dogs. STUDY DESIGN: Retrospective study. ANIMALS: Dogs (n=20) with HAVF. METHODS: Medical records of dogs with HAVF were reviewed. Referring veterinarians and owners were contacted by telephone. History, clinical signs, biochemical and hematologic variables, ultrasonographic and angiographic findings, surgical findings, techniques used to correct the HAVF, survival time, and clinical follow-up were recorded. RESULTS: Canine HAVF often appeared to be an arteriovenous malformation rather than a single fistula. Multiple extrahepatic portosystemic shunts were identified in 19 dogs. Surgery (lobectomy or ligation of the nutrient artery) and/or interventional radiology (glue embolization of the abnormal arterial vessels) was performed in 17 dogs. Thirteen dogs were treated by surgery alone, 4 dogs by glue embolization alone, and 1 dog by glue embolization and surgery. Three dogs treated by surgery alone died <1 month later, and 3 dogs were subsequently euthanatized or died because of persistent clinical signs. None of the dogs treated by glue embolization died <1month after the procedure and all were alive, without clinical signs, at follow-up (9-17 months). Overall, 9 of 12 (75%) dogs with long-term follow-up required dietary or medical management of clinical signs. CONCLUSION: HAVF-related death occurred less frequently after glue embolization than after surgery. CLINICAL RELEVANCE: Glue embolization may be a good alternative to surgery for treatment of certain canine HAVF. PMID- 17461944 TI - Ventroaxial luxation of the apex of the corniculate process of the arytenoid cartilage in resting horses during induced swallowing or nasal occlusion. AB - OBJECTIVE: To report ventroaxial luxation of the apex of the left or right corniculate process of the arytenoid cartilage under the contralateral corniculate process during resting endoscopic examination, and morphologic features of the larynx of 1 affected horse. STUDY DESIGN: Retrospective study. ANIMALS: Horses (n=8). METHODS: Horses had endoscopic examination as part of a survey of Clydesdale horses (n=7), or investigation of poor performance in Thoroughbred horses (1). One Clydesdale was euthanatized and the larynx examined; 4 cadaver larynges from normal horses were also examined. RESULTS: Ventroaxial luxation of the apex of the left or right corniculate process of the arytenoid cartilage was not detected during quiet breathing but was induced by swallowing or nasal occlusion. Prevalence in Clydesdales was 5.2% (7/133). A Thoroughbred with identical endoscopic appearance of the larynx at rest had progressive ventroaxial luxation of the apex of the arytenoid cartilage during high-speed treadmill endoscopy, associated with abnormal respiratory noise. Necropsy examination of an affected Clydesdale larynx revealed an excessively wide (10 mm) transverse arytenoid ligament that allowed easy separation of the apices of the corniculate processes. In normal cadaver larynges, the apices could not be separated with abaxial traction. CLINICAL RELEVANCE: The clinical relevance of this laryngeal observation in resting horses is unclear. Ventroaxial luxation of the corniculate process of the arytenoid cartilage during induced swallowing or nasal occlusion in resting horses or during high-speed treadmill exercise may be caused by an abnormally wide transverse arytenoid ligament. PMID- 17461945 TI - The effect of lidocaine on postoperative jejunal motility in normal horses. AB - OBJECTIVE: To measure the effect of lidocaine on the duration of the migrating myoelectric complex (MMC) and Phases I, II, and III of the MMC, spiking activity of the jejunum, and number of Phase III events when administered postoperatively to normal horses. STUDY DESIGN: Nonrandomized cross-over design. METHODS: Horses were anesthetized and via flank laparotomy 4 silver-silver chloride bipolar electrodes were sutured to the proximal jejunum. Electrical activity was recorded for 6 hours during 3 recording sessions beginning 24, 48, and 72 hours postoperatively. Saline (0.9% NaCl) solution was administered for 3 hours followed by lidocaine administration for 3 hours (1.3 mg/kg bolus intravenously [IV], 0.05 mg/kg/min IV constant rate infusion). RESULTS: Duration of MMC was unchanged during lidocaine administration (77 minutes-saline versus 105 minutes lidocaine, P=.16). Durations of Phase I and II were unchanged during lidocaine administration (P=.19 and .056, respectively). Phase III was shorter during lidocaine administration (P=.002). Spiking activity was unchanged at all time periods during lidocaine administration (24 hours-P=.10; 48 hours-P=.95; and 72 hours-P=.12). The number of Phase III events was unchanged over all time periods during lidocaine administration (P=.053). CONCLUSIONS: Duration of MMC, spiking activity, and number of Phase III events was unchanged during lidocaine administration. CLINICAL RELEVANCE: Use of lidocaine as a prokinetic agent cannot be supported by this study in normal horses; however, results may differ in clinically affected horses. PMID- 17461946 TI - Cytokine and chemokine gene expression of IL-1beta stimulated equine articular chondrocytes. AB - OBJECTIVE: To evaluate mRNA expression of several proinflammatory and anti inflammatory cytokines and chemokines in equine unstimulated and interleukin 1beta (IL-1beta)-stimulated chondrocytes. STUDY DESIGN: In vitro experiment using equine chondrocyte cultures. SAMPLE POPULATION: Whole articular cartilage from metacarpophalangeal joints (n=5 horses; 10 fetlocks). METHODS: Chondrocyte monolayer cultures were established from digested adult equine articular cartilage and stimulated with 5 ng/mL of recombinant human IL-1beta. RNA was extracted from the cells 24 hours after stimulation. IL-1beta, IL-4, IL-6, IL-8, tumor necrosis factor-alpha (TNF-alpha), and ubiquitin (house keeping gene) mRNA expression were investigated by real-time RT-PCR. RESULTS: IL-1beta, IL-6, and IL 8 mRNA were expressed in unstimulated chondrocytes from macroscopically normal joints and were significantly up-regulated after stimulation (5/5 horses). IL-4 mRNA was not detected in any samples (0/5 horses). TNF-alpha mRNA, by comparison, was expressed in 2/5 unstimulated samples and in all stimulated samples but a considerable sample variation in response to IL-1beta stimulation was observed. CONCLUSIONS: Equine chondrocytes express mRNA for several proinflammatory cytokines and chemokines and IL-1beta modulates their expression. CLINICAL RELEVANCE: Chondrocytes express proinflammatory cytokines and chemokines capable of modulating a local inflammatory cascade in articular cartilage, which could potentially lead to focal degradation and osteoarthritis. PMID- 17461947 TI - Does use of a jig influence the precision of tibial plateau leveling osteotomy surgery? AB - OBJECTIVE: To compare the precision of tibial plateau leveling osteotomy (TPLO) surgery when performed with or without a jig. STUDY DESIGN: Cadaveric study. SAMPLE POPULATION: Cadaveric, adult greyhound hindlimbs (n=9 pair). METHODS: TPLO (n=18) was performed on each limb pair; 1 with and 1 without use of a jig. Measurements taken from pre- and postsurgical radiographs were osteotomy position, tibial plateau angle (TPA), varus-valgus malalignment, and tibial torsion. The null hypothesis was that TPLO precision was not affected by use of a jig. Student's t-test was used to investigate differences in TPA, osteotomy location, and frequency and extent of iatrogenic limb malalignment; P<.05 was considered significant. RESULTS: There were no significant differences in postsurgical TPA, tibial crest thickness, varus-valgus malalignment, or tibial torsion between TPLOs performed with or without a jig. Osteotomy location was significantly more distal when a jig was used (P=.03). CONCLUSIONS: Jig use did not improve the precision of TPLO surgery. CLINICAL RELEVANCE: Performing TPLO without a jig should reduce surgery time, eliminate complications related to placement of the distal jig pin and allow unhindered positioning of the osteotomy. PMID- 17461948 TI - Evaluation of pentosan polysulfate sodium in the postoperative recovery from cranial cruciate injury in dogs: a randomized, placebo-controlled clinical trial. AB - OBJECTIVE: To evaluate the efficacy of pentosan polysulfate (PPS) for improving the recovery period and mitigate the progression of osteoarthritis (OA) of the canine stifle after extracapsular stabilization of cranial cruciate ligament (CCL) injuries. STUDY DESIGN: Randomized, blinded, placebo-controlled clinical trial. ANIMALS: Dogs (n=40) with unilateral CCL instability. METHODS: Each dog had an extracapsular stabilization of the stifle with or without partial meniscectomy. Dogs were divided into 4 groups based on preoperative radiographic assessment and whether a partial meniscectomy was performed. Dogs were randomly assigned to either (3 mg/kg) PPS or placebo treatment in each group, and then injected subcutaneously weekly for 4 weeks. Lameness, radiographic changes, biological marker concentration in blood and urine, and ground reaction forces (GRFs) were collected preoperatively, and at 6, 12, 24, and 48 weeks. Data were analyzed within and between groups using repeated measures ANOVA; P<.05 was considered significant. RESULTS: No adverse reactions to PPS were reported. Thirty-nine dogs completed a minimum of 24-weeks follow-up and 33 dogs completed 48 weeks. All dogs clinically improved after surgery without differences in lameness score, vertical GRFs, or radiographic progression. Grouped and evaluated only by initial radiographic score, PPS-treated dogs improved significantly faster in braking GRFs than placebo-treated dogs. In dogs with partial meniscectomies, urine deoxypyridinoline, and serum carboxy-propeptide of type II collagen were significantly increased at 6 weeks in placebo-treated dogs compared with PPS-treated dogs. CONCLUSIONS: PPS administered after stabilization of the cruciate deficient stifle may prove to be a useful adjunctive treatment option, although further studies are necessary to substantiate this claim. PMID- 17461949 TI - Relationship of tibial plateau slope to limb function in dogs treated with a lateral suture technique for stabilization of cranial cruciate ligament deficient stifles. AB - OBJECTIVE: To evaluate the effect of a lateral suture technique (LST) on tibial plateau angle (TPA) measurement and to compare TPA with functional outcome in dogs treated for cranial cruciate ligament (CrCL) rupture with LST. STUDY DESIGN: Prospective clinical study. SAMPLE POPULATION: Dogs (n=34) with unilateral CrCL instability. METHODS: All dogs had lameness examination, survey stifle radiographs, and force plate analysis before and at 6, 12, 24, and 48 weeks after surgery. Radiographic osteoarthritis (OA) scores and lameness scores were assigned using previously reported methods. Preoperative radiographs were performed in all dogs, and postoperative serial radiographs were performed in 6 dogs for measurement of TPA. Differences in TPA measurements were evaluated with a random effects repeated measures model. The significance of LST on TPA measurement was established in 6 dogs and the effect of TPA on vertical impulse, peak vertical force, progression of radiographic scores, and lameness score were analyzed by general linear models in all dogs. Differences were considered significant if P<.05. RESULTS: Significant differences were not noted between pre and serial postoperative measurements of TPA. A significant correlation was not established between TPA and postoperative vertical impulse, peak vertical force, lameness score, or radiographic OA scores. CONCLUSIONS: TPA values were unchanged after LST and TPA does not affect outcome measures in dogs treated with LST. CLINICAL RELEVANCE: TPA has no predictive value on clinical outcome in dogs treated with LST for stabilization of CrCL deficient stifles. PMID- 17461950 TI - Tilt table recovery of horses after orthopedic surgery: fifty-four cases (1994 2005). AB - OBJECTIVE: To describe an assisted anesthetic recovery technique using a tilt table for horses after high-risk orthopedic-related procedures and to evaluate outcome. STUDY DESIGN: Retrospective study. SAMPLE POPULATION: Anesthetic recoveries (n=54) for 36 horses. METHODS: Medical records (April 1994-October 2005) for horses that had high-risk orthopedic surgery and recovery from general anesthesia on a tilt table were reviewed. Information about the surgical procedure, anesthetic and recovery period was analyzed. RESULTS: Of 54 anesthetic recoveries, 1 horse (1.9%) had complete failure of internal fixation during recovery and was euthanatized. Six (11% recoveries) horses failed to adapt to the tilt table system, which necessitated transfer to a conventional recovery room. Complications without important consequences for clinical outcome (skin abrasions, myositis, cast breakage, partial implant failure) occurred during 8 (15%) recoveries. CONCLUSIONS: A tilt table recovery system was associated with minimal incidence of serious complications. Potential disadvantages of the system are the number of personnel required, longer recovery time, and the need for a specialized table. CLINICAL RELEVANCE: A tilt table is a useful system for recovering horses believed to be at increased risk of injury during anesthetic recovery after high-risk orthopedic-related procedures. PMID- 17461951 TI - Rectus abdominis free muscle flap for reconstruction in nine dogs. AB - OBJECTIVE: To evaluate the clinical use and outcome of a rectus abdominis microvascular free flap for wound closure in dogs. STUDY DESIGN: Retrospective case series. ANIMALS: Dogs (n=9) with complex extremity or oral wounds. METHODS: Medical records (2002-2006) of dogs that had a rectus abdominis free tissue transfer to close an extremity or oral wound were reviewed. RESULTS: Nine dogs were identified: 5 had distal extremity wounds, 3 had oral palatal defects, and 1 had a large hygroma excised. A rectus abdominis free tissue transfer with a caudal epigastric vascular pedicle was successfully used for management of these wounds. No major complications occurred with the donor site and a good cosmetic and functional outcome occurred in all dogs. CONCLUSION: The rectus abdominis is a versatile muscle that can be used for reconstructing cutaneous and oral defects with repeatable success. CLINICAL RELEVANCE: Free tissue transfer of the rectus abdominis muscle is a clinically useful technique for closure of a variety of difficult soft tissue wounds. PMID- 17461952 TI - Clinical outcome of nonnasal chondrosarcoma in dogs: thirty-one cases (1986 2003). AB - OBJECTIVE: To evaluate metastatic rate and survival times of dogs with chondrosarcoma of nonnasal bony sites treated by wide surgical excision. STUDY DESIGN: Retrospective study. ANIMALS: Dogs (n=31) with chondrosarcoma. METHODS: Medical records were retrospectively reviewed to identify dogs with chondrosarcoma of bone in potentially surgically accessible sites. When complete information was not available in the medical record, owners and referring veterinarians were contacted by telephone to evaluate the course of disease and survival times. When possible, histopathologic diagnosis was confirmed by a single board certified pathologist and tumors were histologically graded. RESULTS: Dogs treated by wide surgical excision (n=18) had a mean survival time of 3097 days and did not reach median survival time. Dogs untreated except for diagnostic biopsy (n =13) had a median survival time of 523 days and a mean survival time of 495 days. Method of treatment and tumor grade predicted survival time (P=.016 and P=.007, respectively). Metastatic rate was 28% for treated dogs and 15% for untreated dogs, with no significant difference between the 2 groups (P=.39). CONCLUSIONS: Wide surgical excision significantly improves survival time for dogs with chondrosarcoma of nonnasal bony sites, but does not affect the likelihood of metastasis. Grade may be prognostic for survival. CLINICAL RELEVANCE: Surgical excision benefits dogs with chondrosarcoma and can result in prolonged survival times. Metastasis still occurs in approximately 1 of 4 dogs even after surgical resection. PMID- 17461953 TI - In vitro elution studies of amikacin and cefazolin from polymethylmethacrylate. AB - OBJECTIVE: To compare the in vitro elution characteristics of amikacin and cefazolin from polymethylmethacrylate (PMMA) alone and in combination. STUDY DESIGN: A prospective, controlled, experimental study. METHODS: Three aliquots of 6 g sterile PMMA were measured and to them added (1) 750 mg amikacin; (2) 1050 mg cefazolin; and (3) 750 mg amikacin and 1050 mg cefazolin. Ten beads of each antimicrobial/PMMA combination were placed in 5 mL phosphate-buffered saline (PBS) at pH 7.4 and room temperature with constant agitation. PBS was sampled at 15 time points between 1 hour and 30 days. Amikacin concentrations were determined by fluorescence polarization immunoassay and cefazolin concentrations by high-performance liquid chromatography. RESULTS: Amikacin and cefazolin eluted at concentrations greater than 8 and 4 times, respectively, above the minimum inhibitory concentration (MIC) for susceptible bacteria over 30 days. Co-elution of the antibiotics resulted in a greater rate and proportion of antibiotic eluted. Concentrations of amikacin and cefazolin in the co-eluted fluid were not maintained sufficiently above the MIC for selected bacteria over 30 days. CONCLUSIONS: PMMA beads of only amikacin or cefazolin-eluted concentrations greater than the MIC for selected bacteria for 30 days. Co-elution of the antibiotics at the selected doses resulted in a significantly shorter duration of elution and may not be effective for treatment of wound infection. CLINICAL RELEVANCE: Co-elution of amikacin and cefazolin from PMMA at the selected doses cannot be recommended for sustained treatment of infection. PMID- 17461954 TI - Mechanical comparison of an interlocking nail locked with conventional bolts to extended bolts connected with a type-IA external skeletal fixator in a tibial fracture model. AB - OBJECTIVE: To compare the structural properties of interlocking nails (ILNs) locked with bolts (ILNb) to ILN locked with extended bolts connected with a type IA external skeletal fixator (ILN-ESF) in a fracture gap model. STUDY DESIGN: Experimental study. SAMPLE POPULATION: Synthetic tibial bone substitutes. METHODS: Custom-made synthetic tibial bone substitutes were implanted with standard ILNs locked with either bolts or extended bolts connected to an external skeletal fixation (ESF). Constructs were tested in torsion, bending, and axial compression (n=4/testing mode). Data, consisting of construct compliance and associated deformation, were compared using t-tests. RESULTS: The ILN-ESF construct compliance and deformation were significantly less than those of the ILNb construct in torsion, bending, and compression (P<.001). Slack was present in the ILNb construct under torsion and bending, but not in the ILN-ESF construct, regardless of testing mode. CONCLUSIONS: Substitution of locking bolts with extended bolts connected to an ESF significantly reduced the construct compliance and overall deformation in torsion, bending, and compression. Furthermore, the inherent slack of the ILNb was eliminated by the use of an ESF in torsion and bending. CLINICAL RELEVANCE: The improvement in structural properties of the ILN-ESF constructs could diminish interfragmentary motion at the fracture site and potentially improve bone healing. PMID- 17461957 TI - Professional oral health care by dental hygienists reduced respiratory infections in elderly persons requiring nursing care. AB - OBJECTIVES: Respiratory infection is a major cause of death in the elderly. We have evaluated the role of professional oral health care (POHC) by dental hygienists in reducing respiratory infections in elderly persons requiring nursing care. METHODS: Two populations of elderly persons, one receiving POHC and one not, were examined to determine numbers of microorganisms, potent pathogens of respiratory infection, enzymatic activity in saliva, fevers, prevalence of fatal aspiration pneumonia and prevalence of influenza. RESULTS: In the first population, we found a high prevalence of potent respiratory pathogens such as Staphylococcus species, Pseudomonas aeruginosa and Candida albicans. Patients who received POHC showed a lower prevalence for these pathogens than those who did not. The ratio of fatal aspiration pneumonia in POHC patients was significantly lower than that in patients without POHC (non-POHC) over a 24-month period (P < 0.05). The prevalence of a fever of 37.8 degrees C or more in POHC patients was significantly lower than that in the non-POHC group (P < 0.05). In the second study population, we investigated the effects of POHC on infection with influenza over a 6-month period. In the POHC group, neuraminidase and trypsin-like protease activities decreased, and one of 98 patients was diagnosed with influenza; whereas, in the non-POHC group, nine of 92 patients were diagnosed with influenza. The relative risk of developing influenza while under POHC was 0.1 (95% CI 0.01-0.81, P = 0.008). CONCLUSION: These results suggest that POHC by dental hygienists is effective in preventing respiratory infections in elderly persons requiring nursing care. PMID- 17461958 TI - Comparison of manual versus sonic and ultrasonic toothbrushes: a review. AB - PURPOSE: This review of the literature intends to evaluate the effect of brushes with high frequency motion when compared with manual toothbrushes regarding the indices of plaque and gingival bleeding. METHODS: Patients presenting gingivitis and/or chronic periodontitis were evaluated in addition to patients having osseointegrated implants and fixed orthodontic appliances. Pertinent literature was reviewed to select articles according to previously defined inclusion criteria. RESULTS: In the assessed studies results showed significant decreases in plaque and gingival indices by utilization of both types of brushes. However, in the selected studies where sonic brushes were tested in orthodontic and dental implant patients there was a more significant decrease in the indices. Furthermore, there was no indication of gingival recession attributed to product use. CONCLUSION: Future studies with a more homogeneous methodology and better experiment designs will be needed. PMID- 17461959 TI - Self-reported oral problems, compared with clinical assessment in an epidemiological study. AB - OBJECTIVES: To study self-reported oral problems in relation to periodontal clinical findings in an epidemiological investigation. MATERIALS AND METHODS: 1676 individuals, 828 men and 848 women, participated. The clinical examination comprised the number of remaining teeth excluding third molars, plaque index (PLI), gingival index (GI), bleeding on probing (BOP%), calculus index (CI) and the number of teeth with pocket depth (PD) > or =5 mm. Participants answered a questionnaire concerning smoking and snuffing habits and described in their own words any oral problems present. Analysis of variance (anova), unpaired t-test and chi-squared test were the statistical methods used. RESULTS: The questionnaire was completed by 99.4% of participants. Problems of the teeth and mouth were reported by 28.9% of respondents comprising 31.8% females and 26.2% males. The difference between the sexes was statistically significant (P < 0.05). Of the reported oral problems, 57.4% was of periodontal origin. The differences between subjects with and without self-reported mobile teeth showed significant clinical outcome in PLI (P < 0.001), CI (P < 0.01), GI (P < 0.001), BOP% (P < 0.01), the number of teeth with PD > or = 5 mm (P < 0.001) and the number of remaining teeth (P < 0.001). The differences between subjects with and without self-reported periodontal disease and with and without bleeding gingiva also showed significant differences in CI (P < 0.05), GI (P < 0.05) and the number of teeth with PD > or = 5 mm (P < 0.05) and in PLI (0.001), CI (P < 0.01), GI (P < 0.001) and BOP% (P < 0.001) respectively. CONCLUSIONS: Present study clearly shows that self-reported periodontal problems were confirmed by clinical assessments. PMID- 17461960 TI - Peri-implant maintenance of immediate function implants: a pilot study comparing hyaluronic acid and chlorhexidine. AB - INTRODUCTION: In implants, maintenance assumes an important role. The role of chlorhexidine (CHX) is well known in maintenance, while only limited evidence exists on the practical use of hyaluronic acid (HA). The objective of this study was to compare the health status of the peri-implant complex (hard and soft tissues surrounding the implant) during the healing period of immediate function implants, using HA or CHX gels in the patient's maintenance protocol. STUDY POPULATION AND METHODOLOGY: Thirty complete edentulous patients, with four immediate function Branemark System implants placed in the mandible (total of 120 implants), were randomly assigned to two groups (HA and CHX) using only these two chemicals in their daily implant self-care. Both groups were followed up for 6 months, with clinical observations on the 10th day, 2 months, 4 months and 6 months post-surgically. RESULTS: During the course of the study, HA and CHX produced good results in maintaining a healthy peri-implant complex in immediate function implants for complete rehabilitations in the edentulous mandible. Statistically significant differences were found in favour of the HA group in the modified bleeding index on the second observation (P = 0.003). The difference was more marked in the axial implants placed in the fifth sextant (P = 0.05). Correlation coefficient between plaque and bleeding index revealed a potentially better result for CHX at 6 months. CONCLUSION: The findings point out the importance of a maintenance protocol in immediate function implants. Both chemicals are valid tools for implant maintenance. The authors suggest that it might be advantageous to administer HA in the first 2 months and CHX between 2 and 6 months. PMID- 17461961 TI - Attitudes to dental hygienists: evaluation of the Dental Hygienist Beliefs Survey in a Swedish population of patients and students. AB - The aim was to evaluate and test the psychometric properties of the Dental Hygienist Beliefs Survey (DHBS) in a Swedish sample of different patient groups and students. It was hypothesized that negative dental hygienist beliefs would discriminate between fearful and non-fearful study groups. The DHBS was distributed together with the revised Dental Beliefs Survey (DBS-R) and the Dental Anxiety Scale (DAS). The study sample included 394 subjects (130 students, 144 general dental patients, 90 periodontal patients and 30 patients on a waiting list for dental fear treatment). The results verified that the DHBS discriminates well between dentally fearful and non-fearful study groups. The DHBS had high internal consistency (Cronbach's alpha = 0.96-0.98) in all the groups. The correlation between the DHBS and the DBS-R was high (rho = 0.82, P < 0.001). Furthermore, the DHBS correlated significantly with the DAS, as well as with a low but significant correlation to age (more negative attitudes in younger age groups) and gender (more negative attitudes amongst women). Regression analysis showed that gender and the DHBS items: 23, 16 and 28, i.e. items related to feeling helpless, worries/fears not being taken seriously and fear about 'bad news' possibly preventing treatment, were the most important predictors of dental fear. The results suggest that the DHBS may be a valid and reliable scale to use in order to assess patient's specific attitudes to dental hygienists. However, the psychometric properties including test-retest analysis and the underlying factor structure of the DHBS need to be further explored. PMID- 17461962 TI - A predictive study on the role and function of the dental hygienist in Taiwan. AB - OBJECTIVES: The purpose of this study was to achieve a consensus on the work roles and scope of practice of dental hygienists in Taiwan and to rank the results according to their importance. METHODS: A modified Delphi technique was used to enquire how experts view their role and function as a dental hygienist in Taiwan. Three groups of 105 experts (dentists, officials and scholars) were surveyed. Responses were analysed using qualitative statistics. RESULTS: Fifty one topics, which were included in the categories of the research agenda, were identified through the first phase of the study. The return rate of 87.62% and 68.48% was achieved from the two rounds of surveys. Afterwards, a consensus was reached on 38 out of the 51 topics. In the aspect of service, the most desired roles and functions were those of a friendly receptionist and of an employee effective in scheduling appointments. In the aspect of administration, the most important roles and functions were billing of health insurance, and book keeping on payments, as well as administering materials. In the aspects of technical efficiency, dental hygienists were most frequently seen as serving in four- or six-handed assisting and in infection control. In the area of oral health, an oral health educator and an oral health promoter were viewed as having the most important roles and functions. CONCLUSIONS: The experts concluded that the role and function of a dental hygienist should shift from a patient treatment orientation to that of an oral health promoter. The role expectations need to change to reflect what hygienists actually learn and do. PMID- 17461963 TI - Antimicrobial activity of garlic against oral streptococci. AB - The antimicrobial activity of two garlic clones' (1: purple and 2: white) crude extracts against oral microbiota was evaluated in vitro (study 1) and in vivo (study 2). Study 1 consisted of the evaluation of minimum inhibitory (MIC) and bactericidal (MBC) concentrations against nine streptococci strains. In study 2, a 2.5% garlic (clone 2) solution was used as a mouthwash in a 5-week study by 30 subjects. Blood agar and Mitis Salivarius Bacitracin agar were inoculated with subjects' saliva to quantify oral microorganisms and mutans streptococci. Study 1 showed MIC ranging from 0.5 to 32.0 mg ml(-1) for clone 2 and from 8 to 64.0 mg ml(-1) for clone 1. MBC ranged from 1.0 to 128.0 mg ml(-1) and from 8.0 to 128.0 mg ml(-1) regarding clones 2 and 1 respectively. Study 2 showed that 2.5% garlic mouthwash solution had good antimicrobial activity against mutans streptococci and oral microorganisms. Maintenance of reduced salivary levels of streptococci was observed after 2 weeks at the end of mouthwash use. Unpleasant taste (100%), halitosis (90%) and nausea (30%) were reported by subjects after the end of the study. It was concluded that the garlic clones have antimicrobial properties in vitro against streptococci and anticariogenic properties against oral microorganism in spite of its adverse effects. PMID- 17461964 TI - Legionella in the dental office. AB - OBJECTIVE: This study aimed to assess the microbiology of dental unit water and municipal water in terms of Legionella species and total bacteria levels. METHODS: The presence of Legionella species was investigated using the culture method, direct fluorescent antibody and polymerase chain reaction techniques in collected dental unit water and municipal water samples from 71 dental offices in Ankara, Turkey. In addition, total bacterial counts were assessed using the culture method. RESULTS: In 27% of the dental unit water samples and in 13% of municipal water samples, the number of colony-forming units (cfu ml(-1)) significantly exceeded acceptable values for high-risk group patients. No Legionella spp. was found in the dental unit water samples. Legionella SG3 was found in only one municipal water sample. CONCLUSION: The dental unit water systems examined in this study did not include Legionella spp., but other bacteria at high numbers were determined. This is a potential threat, especially for elderly people, the medically compromised patients receiving regular dental treatment and the dental clinic staff. PMID- 17461965 TI - Heart disease: the number one killer of women. PMID- 17461966 TI - Organization for Safety and Asepsis Procedures (OSAP). PMID- 17461967 TI - Juvenile periodontitis. PMID- 17461970 TI - Quantitative analysis of RNA-mediated protein-protein interactions in living cells by FRET. AB - Specific assembly of ribonucleoprotein complexes is essential in controlling various cellular functions including gene regulation. Diverse scaffolds containing proteins or nucleic acids could play key roles in stabilizing specific ribonucleoprotein complexes by enhancing protein-protein or RNA-protein interactions. One such example is the assembly of active RNA polymerase II transcription elongation complex originating from HIV-1 long terminal repeat promoter that involves HIV-1-encoded Tat protein and viral mRNA structure, trans activation responsive RNA, and human CyclinT1 which is a subunit of the positive transcription elongation factor complex b. By using genetically encoded fluorescent proteins fused with Tat and human CyclinT1, here we demonstrate that human CyclinT1 was diffused throughout the nucleus and specific interactions between Tat and human CyclinT1 altered the localization of human CyclinT1 to specific nuclear foci. We also found that trans-activation responsive RNA enhanced protein-protein interactions between human CyclinT1 and Tat in living cells. Our results highlights the importance of trans-activation responsive RNA as a scaffold for stable and high affinity assembly of two protein partners to form a regulatory switch essential in HIV-1 gene regulation. RNA-mediated assembly of ribonucleoprotein complexes could be a general mechanism for stable ribonucleoprotein complex formation and a key step in regulating other cellular processes and viral replication. Furthermore, our results suggest that Tat interactions with human CyclinT1 change the nuclear location of positive transcription elongation factor complex b to modulate positive transcription elongation factor complex b function and transcription of cellular genes. PMID- 17461971 TI - alpha(1)-Adrenergic receptor stimulation of cell motility requires phospholipase D-mediated extracellular signal-regulated kinase activation. AB - Phospholipase D is suspected to play a role in tumorigenesis, and the inhibition of phospholipase D has been associated with changes in several cellular events including invasion and migration. We report here that the specific alpha(1) adrenergic receptor agonist, phenylepherine, signals to a growth factor pathway in a manner that requires phospholipase D activity in CCL39 fibroblasts. Phenylepherine increased extracellular signal-regulated kinase phosphorylation eightfold and promoted stress fiber formation threefold. Stress fiber formation was blocked when extracellular signal-regulated kinase activation was inhibited. Stimulation of CCL39 fibroblasts by phenylepherine increased the rate of wound healing fourfold in a wounding assay, while treatment with the MEK inhibitor, PD98059 reduced the closure of phenylepherine-induced wound healing to control levels. Addition of 1-butanol but not 2-butanol inhibited extracellular signal regulated kinase activation by phenylepherine, presumably by blocking the formation of phosphatidic acid. Exogenously added cell permeable phosphatidic acid increased extracellular signal-regulated kinase activation in a time- and dose-dependent manner as well as stimulated the formation of stress fibers. 1 butanol also significantly inhibited the ability of phenylepherine to stimulate stress fiber formation and wound healing. Taken together, these results indicate a novel role for phospholipase D in the activation of the extracellular signal regulated kinase growth factor pathway to stimulate early cellular events induced by phenylepherine. PMID- 17461972 TI - Ab initio study of non-peptidic antihypertensives. AB - In this study, the authors report ab initio molecular orbital calculations on natural hormone angiotensin II (ANG II) that induces activity at AT(1) receptor leading to vasoconstriction and subsequent hypertension. Optimized conformations and charge distributions of various conformers of natural hormone and AT(1) antagonists have been studied. The major pharmacophoric features have been deduced. The charge environment of ANG II and drugs guided us in exploring the two possibilities: substrate inhibition and competitive inhibition. The results indicate that more potent drugs avoid 'wastage' in substrate inhibition and undergo strong competitive antagonism at the receptor. Specific binding interactions are essential for competitive antagonism. Slight differences in conformation may effect to differences in interactions with the receptor, hence modulating the antagonistic properties of the drug. PMID- 17461973 TI - Uncovering genetic relationships using small molecules that selectively target yeast cell cycle mutants. AB - Genetic analysis in budding yeast has shown that multiple G1 cyclins and cyclin dependent kinases control cell cycle entry, polarized growth, and spindle pole duplication. The G1 cyclins Cln1 and Cln2 associate with the cyclin-dependent kinase Cdc28 to facilitate cell cycle progression and development of the cleavage apparatus. We have developed a chemical genetic approach toward the discovery of compounds that target G1 control pathways by screening for compounds that selectively kill a yeast strain lacking the G1 cyclins Cln1 and Cln2. A class of small molecules was identified that is highly toxic toward the cln1 Delta cln2 Delta double mutant and has relatively little effect on wild-type yeast. We call these compounds 'clinostatins' for their selectivity toward the cln1/2 deletion strain. Clinostatins were used in a genome-wide chemical synthetic lethality screen to identify other genes required for growth in the presence of the drug. Other deletions that were sensitive to the drug include members of the protein kinase C(PKC)-dependent MAP kinase pathway. These results suggest an approach for combining chemical synthetic lethality and chemical genomic screens to uncover novel genetic interactions that can be applied to other eukaryotic pathways of interest. PMID- 17461974 TI - Synthesis and in vitro evaluation of substituted phenyl-piperazinyl-phenyl oxazolidinones against Gram-positive bacteria. AB - With the incidence of linezolid-resistant Enterococcus faecalis, E. faecium and Staphylococcus aureus, modification of linezolid at the 5- and/or 3-positions led to the development of a series of 3-(methoxyl-phenyl)-piperazinyl-phenyl oxazolidinone analogues. These compounds were tested in vitro against six gram positive standard organisms (S. aureus, S. epidermidis, S. pneumoniae, S. albus, Streptococcus enteridis and S. nonhemolyticus). 5-acetylaminomethyl oxazolidinones bearing fluorine at 3'-position of phenyl ring showed activities against several gram-positive bacteria (MIC: 3.13-6.25 mug/mL). The position of methoxyl group on the phenyl ring of piperazine group affected antibacterial spectrum. 3-(4'- (para-methoxyl-phenyl)-piperazinyl)-(3'-fluoro)-phenyl-5 acetylaminomethyl oxazolidinone was found active against 5 gram-positive organisms except S. nonhemolyticus, whereas 3-(4'-(ortho-methoxyl-phenyl) piperazinyl)-(3'-fluoro)-phenyl-5-acetylaminomethyl oxazolidinone was found active only against 2 gram-positive organisms, namely S. albus, S. enteridis. PMID- 17461975 TI - In silico prediction of SARS protease inhibitors by virtual high throughput screening. AB - A structure-based in silico virtual drug discovery procedure was assessed with severe acute respiratory syndrome coronavirus main protease serving as a case study. First, potential compounds were extracted from protein-ligand complexes selected from Protein Data Bank database based on structural similarity to the target protein. Later, the set of compounds was ranked by docking scores using a Electronic High-Throughput Screening flexible docking procedure to select the most promising molecules. The set of best performing compounds was then used for similarity search over the 1 million entries in the Ligand.Info Meta-Database. Selected molecules having close structural relationship to a 2-methyl-2,4 pentanediol may provide candidate lead compounds toward the development of novel allosteric severe acute respiratory syndrome protease inhibitors. PMID- 17461976 TI - Plasmodium and host lactate dehydrogenase molecular function and biological pathways: implication for antimalarial drug discovery. AB - Lactate dehydrogenase is an enzyme that catalyses the interconversion of pyruvate and lactate with concomitant interconversion of NADH and NAD(+). Lactate dehydrogenase is present at high levels in humans and Plasmodium spp. However, the function of lactate dehydrogenase in malarial infection is not well characterized. In this investigation, a new gene ontology technology is used to predict molecular function and biological pathways of lactate dehydrogenase. In comparison with human lactate dehydrogenase, the P. falciparum lactate dehydrogenase has similar molecular functions such as L-lactate dehydrogenase activity. Furthermore, P. falciparum lactate dehydrogenase has L-malate dehydrogenase activity. Although the amino acid sequences for human and P. falciparum lactate dehydrogenase are very different, the molecular functions are similar. This suggests that any non-selective therapeutic treatment aimed at blocking P. falciparum lactate dehydrogenase function may affect human lactate dehydrogenase. In contrast, a selective lactate dehydrogenase inhibitor targeting the l-malate dehydrogenase function of P. falciparum and its corresponding tricarboxylic acid cycle provides an attractive therapeutic opportunity. PMID- 17461979 TI - Molecular evidence-based medicine: evolution and integration of information in the genomic era. AB - Evidence-based medicine and molecular medicine have both been influential in biomedical research in the last 15 years. Despite following largely parallel routes to date, the goals and principles of evidence-based and molecular medicine are complementary and they should be converging. I define molecular evidence based medicine as the study of medical information that makes sense of the advances of molecular biological disciplines and where errors and biases are properly appreciated and placed in context. Biomedical measurement capacity improves very rapidly. The exponentially growing mass of hypotheses being tested requires a new approach to both statistical and biological inference. Multidimensional biology requires careful exact replication of research findings, but indirect corroboration is often all that is achieved at best. Besides random error, bias remains a major threat. It is often difficult to separate bias from the spirit of scientific inquiry to force data into coherent and 'significant' biological stories. Transparency and public availability of protocols, data, analyses and results may be crucial to make sense of the complex biology of human disease and avoid being flooded by spurious research findings. Research efforts should be integrated across teams in an open, sharing environment. Most research in the future may be designed, performed, and integrated in the public cyberspace. PMID- 17461980 TI - Cardiac biomarkers and survival in haemodialysis patients. AB - BACKGROUND: In dialysis patients, cardiac troponin T (cTNT) is often elevated despite the absence of acute myocardial ischaemia, and amino-terminal pro-B natriuretic peptide (NT-proBNP) is markedly higher compared to non-haemodialysis patients. In a longitudinal observation, we evaluated the association of cTNT and NT-proBNP on cardiovascular morbidity and mortality in haemodialysis patients with and without fluid overload. MATERIALS AND METHODS: Plasma cTNT levels of 134 haemodialysis patients were measured before and after a dialysis session by 3rd generation electro-chemoluminiscence immunoassay. NT-proBNP was determined using a polyclonal antibody recognizing the N-terminal fragment of BNP (Elecsys autoanalyzer 2010, Roche Diagnostics, Mannheim, Germany). Volume status was determined by a clinical score system. Cardiovascular morbidity and mortality were assessed over a follow-up period of 36 months. RESULTS: Plasma cTNT > 0.03 ng mL(-1) was found in 39.6% of all patients. Patients with hypervolaemia had significantly higher cTNT levels compared to euvolaemic patients (median 0.054 ng mL(-1), interquartile range 0.019-0.153 vs. 0.005 ng mL(-1), < 0.001-0.034; P < 0.001). All haemodialysis patients had excessively high levels of NT-proBNP (median 4524; interquartile range 2000-10 250 pg mL(-1)), and NT-proBNP was significantly higher in hypervolaemic haemodialysis patients (11 988, 5307-19 242) compared to euvolaemic haemodialysis patients (3247, 1619-5574); P < 0.001. Receiver operator curves showed a threshold of cTNT > 0.026 ng mL(-1) and NT proBNP > 5300 pg mL(-1) as predictors of hypervolaemia. Asymptomatic chronic haemodialysis patients with cTNT > 0.026 ng mL(-1) and NT-proBNP > 5300 pg mL(-1) were more likely to die due to cardiac events in the follow-up period. Multivariate analysis documented that elevated cTNT and NT-proBNP levels were highly predictive for cardiovascular events. CONCLUSIONS: Plasma levels of cTNT are elevated in approximately 40% and NT-proBNP levels in 100% of asymptomatic chronic haemodialysis patients. Both parameters depend on volume status. Increased NT-proBNP and cTNT are strongly associated with adverse outcome in end stage renal disease patients undergoing haemodialysis, and are a useful tool for risk stratification in chronic haemodialysis patients. PMID- 17461981 TI - Co-administration of ezetimibe and simvastatin in acute myocardial infarction. AB - BACKGROUND: Recent trials in acute myocardial infarction indicate that intensive and early statin therapy that lowers low-density lipoprotein cholesterol (LDL-C) to < or = 70 mg dL(-1) is beneficial. The combination of statins with ezetimibe, a newly developed cholesterol-absorption inhibitor, can lead to a further reduction in LDL-C of up to 26%. In this study, we examined the rapidity and intensity of the lipid-lowering effect of ezetimibe co-administered with simvastatin immediately after myocardial infarction. MATERIALS AND METHODS: Sixty patients admitted for acute myocardial infarction were randomized to receive either simvastatin 40 mg (SIMVA), a combination of simvastatin 40 mg and ezetimibe 10 mg (EZE/SIMVA), or no lipid-lowering drugs (NLLD) and had their lipid levels assessed 2, 4 and 7 days later. RESULTS: At baseline, cardiovascular risk factors were similar in all three groups [mean (SD) LDL-C of 141 (36) mg dL( 1)]. At days 2 , 4 and 7 there was no significant change in mean LDL-C levels in the NLLD group (-10%, -6%, and -9%, all P > 0.09), while there were significant reductions with SIMVA (-15%, -27%, and -25%, respectively, all P < 0.001 vs. day 0) and even greater reductions with co-administration of EZE/SIMVA (-27%, -41%, and -51%, respectively, all P < 0.001 vs. day 0). The percentages of patients achieving LDL-C below 70 mg dL(-1) at days 4 and 7 were substantially greater with EZE/SIMVA (45% and 55%, respectively) than with SIMVA (5% and 10%, respectively), while no NLLD patient reached this goal. Triglyceride levels showed a progressive increase in the NLLD group (+45% at day 7, P < 0.05 vs. day 0), no change in the SIMVA group, but a decrease in the EZE/SIMVA group (-17% at day 7, P < 0.05 vs. day 0). No significant difference in HDL-C levels, tolerability, or clinical events was observed between the three groups. CONCLUSIONS: The co-administration of ezetimibe 10 mg with simvastatin 40 mg, by inhibiting cholesterol absorption and production, allowed more patients with acute myocardial infarction to reach LDL-C < or = 70 mg dL(-1) as early as the fourth day of treatment. The effects of such rapid and intense reduction in LDL-C on cardiovascular morbidity and mortality need to be evaluated in future clinical endpoint studies. PMID- 17461982 TI - Biological variation of asymmetric dimethylarginine and related arginine metabolites and analytical performance goals for their measurement in human plasma. AB - BACKGROUND: Asymmetric dimethylarginine (ADMA) is an endogenous competitive inhibitor of nitric oxide synthase which is believed to be a cause of endothelial dysfunction and has been shown to predict the occurrence of acute coronary events. Data regarding the biological variation of arginine and its methylated derivatives are conspicuously absent from the literature. Such data are important in setting analytical quality specifications, assessing the utility of population reference intervals and assessing the significance of changes in serial results from an individual. MATERIALS AND METHODS: Arginine, homoarginine, ADMA and symmetric dimethylarginine (SDMA) are measured in plasma by high performance liquid chromatography. Twelve healthy volunteers underwent weekly blood sampling for 20 weeks in order to determine the intra- and inter-individual biological variation of these analytes, from which analytical quality specifications, indices of individuality (II) and reference change values (RCV) are derived. Plasma samples from 100 healthy individuals were obtained in order to determine population reference intervals. RESULTS: ADMA and symmetric dimethylarginine (SDMA) exhibit low intra-individual biological variation of 7.4% and 5.8%, respectively, imposing desirable imprecision goals (CV(A)) of < or = 3.7% and 2.9% for these analytes. The described methodology achieves these goals, with analytical CVs of < 3.5% for all analytes. Goals for bias and total error were 3.1-10.1% and 7.2-16.0%, respectively. Reference intervals for ADMA and SDMA were 0.29-0.63 micromol L(-1) and 0.24-0.55 micromol L(-1), but have IIs < 1. RCVs were at least 20% for all analytes studied. CONCLUSIONS: Dimethylarginine concentrations are tightly controlled in health, with the result that imprecision goals for laboratory methods require to be low. Relatively large differences are required between serial results to denote a significant change. Population reference intervals for dimethylarginines are likely to be of limited value in detecting 'abnormality' in an individual from a single result. PMID- 17461983 TI - Elevated levels of interleukin-1beta-converting enzyme and caspase-cleaved cytokeratin-18 in acute myocardial infarction. AB - BACKGROUND: Systemic inflammation and apoptosis-specific immune activation play a major role in acute coronary syndromes (ACS) including acute myocardial infarction (AMI). The role of systemic and coronary obtained inflammatory plasma protein interleukin-1beta precursor (IL-1betap), IL-1beta-converting enzyme (ICE) and the apoptosis-specific caspase-cleaved cytokeratin-18 (ccCK-18) are not known in ACS. MATERIALS AND METHODS: Plasma samples were obtained from stable angina (SA, n = 34), unstable angina (UA, n = 37) and patients with AMI (n = 39). Coronary blood was acquired by means of thrombectomy devices (X-sizer) in AMI patients. IL-1betap, ICE and ccCK-18 were determined by enzyme-linked immunosorbent assay (ELISA). Group comparisons were evaluated by parametric Tukey test. Multivariate logistic regression analysis was performed to determine predictive values of IL-1betap, ICE and ccCK-18 as compared to creatine kinase (CK) and troponin T (TnT) in order to relate these markers with the occurrence of myocardial damage. RESULTS: IL-1betap, ICE and ccCK-18 were identified to be significantly altered in the peripheral blood of patients suffering from AMI as compared to SA and UA. ROC curves were plotted and revealed that ccCK-18 is a novel sensitive marker for the detection of myocardial damage as compared to TnT or CK. (AUC ccCK-18 0.925, TnT AUC 0.62 and CK AUC 0.858.) Moreover, ICE and ccCK 18 were significantly increased at the site of coronary occlusion as compared to peripheral blood samples in AMI patients (both P < 0.001). CONCLUSION: Our data suggest that ACS is related to increased concentration of systemic soluble ICE and ccCK-18. Moreover, soluble ccCK-18 was identified to be a superior marker as compared to TnT or CK, for detection of myocardial damage. PMID- 17461984 TI - Endothelial function in familial combined hyperlipidaemia. AB - BACKGROUND: Familial combined hyperlipidaemia (FCH) is characterized by dyslipidaemia, visceral obesity and insulin resistance, and is associated with an increased intima-media thickness (IMT) and an increased risk for cardiovascular disease. In the present study, we investigated whether FCH is associated with early functional vascular wall changes, as represented by endothelial dysfunction, and we determined whether endothelial function in FCH is related to any of the cardiovascular risk factors associated with the FCH phenotype, or to the (increased) IMT. DESIGN: In 98 patients with FCH [mean age 51 (48-54) years, 43% male] and 230 unaffected relatives [mean age 44 (42-46) years, 48% male], venous blood was drawn in the fasting state after discontinuation of lipid lowering drugs for at least 4 weeks (if used). IMT was measured by B-mode ultrasound and endothelial function was assessed by determination of flow mediated dilation (FMD) and by measurement of plasma concentrations of various soluble adhesion molecules, including soluble vascular cell adhesion molecule (sVCAM), soluble intercellular adhesion molecule (sICAM) and soluble E-selectin. RESULTS: There were no significant differences between FCH patients and their non affected relatives in FMD [2.9 (2.3-3.6%) vs. 2.8 (2.5-3.2%)] or in the plasma concentrations of the various adhesion molecules. None of the individual clinical and biochemical cardiovascular risk factors was an independent predictor of endothelial function in patients with FCH, nor was IMT. However, subgroup analysis revealed that IMT was an independent and powerful predictor of FMD in subjects with carotid artery plaques (St. beta = 4.11, P < 0.004), whereas IMT was no significant predictor in subjects without plaques. CONCLUSIONS: FCH patients have no impaired endothelial function when compared to their unaffected relatives. IMT is an important predictor of FMD when advanced morphological wall changes are present. Our results question the value of FMD measurements for cardiovascular risk stratification in populations with an anticipated high cardiovascular risk. PMID- 17461985 TI - Endothelial dysfunction in a murine model of thyroid hormone resistance. AB - BACKGROUND: The development of a knockin mouse model of resistance to thyroid hormone (RTH) has led to a greater understanding of both the molecular and clinical behaviour of this syndrome. We have investigated the vascular response in RTH using a specific (TRbeta PV) knockin mouse model targeting the PV mutation to the thyroid hormone receptor beta gene locus. MATERIALS AND METHODS: Ring segments of the thoracic aorta were used to assess the response of homozygous, heterozygous and wild-type controls to contractile agents, potassium chloride and phenylephrine. Each genotype after maximal contraction was exposed to increasing concentrations of relaxing agents, acetylcholine (ACh) and sodium nitroprusside (SNP). RESULTS: The response of these aortic ring segments to ACh and SNP demonstrates that endothelium-dependent relaxation to ACh was significantly impaired in both heterozygous and homozygous mice compared to controls (69.8 +/- 2.0%, 59.7 +/- 1.4% and 75.0 +/- 1.7%, respectively; P < 0.001). However, endothelium independent responses to SNP showed no difference between genotypes (114.4 +/- 3.2%, 116.8 +/- 2.6% and 106.9 +/- 4.9%; P = NS). CONCLUSION: These data suggest that endothelial function is impaired in the RTH mouse aorta. The respective roles of elevated thyroid stimulating hormone (TSH), elevated thyroid hormone concentrations and the mutated thyroid hormone beta receptor require further elucidation. PMID- 17461986 TI - Development of a heterologous, multigenotype vaccine against hepatitis C virus infection. AB - BACKGROUND: Unquestionably viral diversity and genetic heterogeneity in hepatitis C virus (HCV) infection and other viral diseases play an essential role in viral immune escape and the development of chronicity. Despite this knowledge most vaccine approaches against HCV have excluded this important issue. Moreover the feasibility of developing an effective HCV vaccine has been questioned, mainly because prophylactic immunity against HCV cannot be achieved in chimpanzees by either vaccination or previous HCV infection, and reinfection in men has been reported, most likely due to genetic shift and immune escape. To analyse and characterize a new technique of a 'multigenotype'- and/or 'library'-vaccine, we established an envelope 1 (E1) plasmid vaccine against HCV and characterized humoral and cellular immune responses after vaccination in a mouse model. MATERIAL AND METHODS: Normally genetic information of one or two target proteins is cloned into a DNA-vaccine. In our approach we cloned a defined number of different genotypes and subtypes (defined vaccine, DV) or the genetic information from 20 patients (undefined) into a plasmid (library vaccine, LV). RESULTS: As expected, immunized animals showed both stronger humoral (ELISA) and cellular (T cell proliferation, ELISPOT) immune responses against genotype 1, since the stimulating antigen was genotype 1 derived. However, not all genotype 1 immunized animals recognized this viral antigen leading to the assumption that some epitopes lost their immunogenicity through a change in the amino acid sequence. Interestingly, some of the genotype 4 and 5 immunized mice sera were able to react against E1 protein. CONCLUSION: Most of the assays showed immune reactivity against the DV or LV vaccine demonstrating the cross-reactive potential of such a vaccination approach. This cloning and immunization strategy based on the viral heterogeneity of the virus has in our view major implications for HCV, a virus with a broad viral genetic diversity, and may become in the future in the context of DNA- or viral-based vaccination strategies a possibility to overcome viral immune escape both in the prophylactic or therapeutic setting. PMID- 17461987 TI - Alteration of bone cell function by RANKL and OPG in different in vitro models. AB - BACKGROUND: Receptor activator of nuclear factor-kappaB ligand (RANKL) and osteoprotegerin (OPG) are well-documented potent regulators of osteoclast development. However, their effects in mature bone cells and in organ cultures have not been well studied. It is uncertain whether their activities in different experimental models are comparable. MATERIALS AND METHODS: RANKL and OPG were evaluated for their activities in mouse calvarial organ cultures, mouse bone marrow cultures, isolated rat mature osteoclast assays and rat primary osteoblast cultures. Results In murine calvarial organ culture, both muRANKL (> or = 10 ng mL(-1)) and rRANKL (> or = 100 ng mL(-1)) significantly stimulated (45)Ca release, while OPG (> or = 50 ng mL(-1)) was an inhibitor of bone resorption. Meanwhile, [(3)H]-thymidine incorporation in this assay was also modulated (indicating proliferation increases in the osteoblast lineage of cells) although these peptides had no direct effect on [(3)H]-thymidine incorporation in isolated osteoblast assays. In mouse bone marrow cultures, muRANKL (> or = 1 ng mL(-1)) and rRANKL (> or = 5 ng mL(-1)) significantly stimulated osteoclastogenesis. The number of nuclei per osteoclast was also significantly increased. OPG strongly inhibited this index, with over 90% suppression at 1 ng mL(-1). Both muRANKL (10 ng mL(-1)) and rRANKL (100 ng mL(-1)) stimulated, while OPG (10 ng mL(-1)) inhibited osteoclast activity in isolated mature osteoclast assays. CONCLUSION: The current study demonstrated that bone resorption modulated by RANKL and OPG, in murine calvarial organ culture, leads to changes in osteoblast proliferation, suggesting a feedback mechanism from osteoclasts to osteoblasts. In addition, it was found that RANKL and OPG have more potent effects on osteoclastogenesis than on the activity of mature osteoclasts. PMID- 17461988 TI - NF-kappaB and apoptosis in colorectal tumourigenesis. AB - BACKGROUND: Nuclear factor-kappaB (NF-kappaB) may play an important role in colorectal tumourigenesis, controlling cell cycle and apoptosis gene expression. In addition, imbalances between cell proliferation and cell death are thought to underlie neoplastic development. The aims of this study were to investigate apoptosis and expression of several apoptosis-related proteins, and to determine correlations with colorectal tumour progression. MATERIALS AND METHODS: Apoptosis was evaluated by the TUNEL assay in 48 patient samples, including adenomas, adenocarcinomas and adjacent normal mucosas. Immunohistochemistry was performed for Bcl-2 and NF-kappaB. Expression levels of p53, Bax and IkappaB proteins were determined by immunoblotting. Cultured human colon cancer cells were used to evaluate NF-kappaB expression and nuclear translocation by immunocytochemistry and immunoblotting. RESULTS: Apoptosis and NF-kappaB immunoreactivity were significantly higher in tumour tissue compared with normal mucosa (P < 0.01), increasing in association with histological tumour progression (P < 0.01). Bcl-2 was consistently higher in normal mucosa (P < 0.01) and inversely correlated with the percentage of apoptosis (P < 0.01). Phosphorylated p53 and Bax levels were similar in tumour tissue and normal mucosa; however, the NF-kappaB inhibitor, IkappaB, tended to decrease in tumours. In vitro, nuclear translocation of NF kappaB was greater in proliferative than in resting phases of colon cancer cells. CONCLUSIONS: NF-kappaB expression and apoptosis are increased from adenoma to poorly differentiated adenocarcinoma tissues. Apoptosis is correlated with suppression of Bcl-2 expression, but appears to proceed through a p53- and Bax independent pathway. Activation of NF-kappaB may play an important role in colorectal tumour progression. PMID- 17461989 TI - The PPARdelta agonist GW501516 suppresses interleukin-6-mediated hepatocyte acute phase reaction via STAT3 inhibition. AB - BACKGROUND: Interleukin-6 and downstream liver effectors acute phase reactants are implicated in the systemic inflammatory reaction. Peroxisome proliferator activated receptor delta (PPARdelta), which binds to and is activated by a variety of fatty acids, was recently shown to have anti-inflammatory actions. MATERIALS AND METHODS: We examined the ability of the synthetic PPARdelta agonist GW501516 to suppress interleukin-6-induced expression of acute phase proteins in human hepatoma HepG2 cells and rat primary hepatocytes. Results GW501516 dose dependently suppressed interleukin-6-induced mRNA expression of the acute phase protein alpha1-antichymotrypsin in HepG2 cells. The compound also suppressed interleukin-6-induced mRNA expression of alpha2-acid glycoprotein, beta fibrinogen and alpha2-macroglobulin in and the secretion of C-reactive protein by rat primary hepatocytes. Depletion of the PPARdelta receptor, but not of PPARalpha or gamma, attenuated the suppressive effect of GW501516 on interleukin 6-induced alpha1-antichymotrypsin mRNA expression, indicating that PPARdelta specifically mediated this effect. Since interleukin-6 stimulates the transcriptional activity of the alpha1-antichymotrypsin promoter by activating the signal transducer and activator of transcription (STAT) 3, we examined functional interaction of this transcription factor and PPARdelta on this promoter. Overexpression of PPARdelta enhanced the suppressive effect of GW501516 on STAT3-activated transcriptional activity of the alpha1-antichymotrypsin promoter, while GW501516 suppressed interleukin-6-induced binding of this transcription factor to this promoter. CONCLUSIONS: These findings indicate that agonist-activated PPARdelta interferes with interleukin-6-induced acute phase reaction in the liver by inhibiting the transcriptional activity of STAT3. PPARdelta agonists might be useful for the suppression of systemic inflammatory reactions in which IL-6 plays a central role. PMID- 17461991 TI - Individualization in the management of fungal disease in the transplant recipient. PMID- 17461992 TI - Posaconazole salvage therapy allows successful allogeneic hematopoietic stem cell transplantation in patients with refractory invasive mold infections. AB - We describe the clinical courses of 3 patients with hematologic malignancies (2 with acute myelogenous leukemia and 1 with multiple myeloma) who developed invasive fungal infections due to uncommon molds (Alternaria spp., Paecilomyces lilacinus, and Zygomycetes). Breakthrough invasive fungal infections of the sinus (n=1), lung (n=3), and pericardium (n=1) developed despite fluconazole prophylaxis and failed to respond to treatment with other licensed antifungal therapies, including amphotericin B (n=3), caspofungin (n=2), and voriconazole (n=3), and surgical intervention (n=2). Salvage therapy with posaconazole oral suspension resulted in successful outcomes in all 3 patients, who subsequently underwent allogeneic hematopoietic stem cell transplantation (HSCT) while on continued posaconazole therapy. The median duration of posaconazole treatment before HSCT was 5 months (range: 1.5-6 months). Posaconazole salvage therapy allowed successful allogeneic HSCT in 3 patients with refractory invasive mold infections. PMID- 17461993 TI - Tuberculosis in renal transplant recipients. AB - Tuberculosis (TB) is a frequent infectious complication in patients on renal replacement therapy, as a result of immunosuppression from uremia and drugs in the post-transplantation period. A retrospective study of all renal transplantation patients from 1989 to date was conducted. This study tried to examine the prevalence, course, and outcome of TB in renal transplant recipients. A comparison with the occurrence of TB in other modalities of renal replacement therapy was also made. We also discussed the treatment protocols for TB in this group of patients. No difference in the prevalence, age, or male/female ratio of TB was seen among the 3 modes of renal replacement therapy. TB of the lung was the more favored site of infection in patients on hemodialysis (77.3%), when compared with those on CAPD (30%) and renal transplant recipients (33.3%). In renal transplant recipients, no deaths occurred due to TB. In 7 patients there was co-infection with cytomegalovirus and in 3 patients there was Aspergillus lung infection. PMID- 17461994 TI - Oral valganciclovir as preemptive therapy for cytomegalovirus infection post allogeneic stem cell transplantation. AB - Antiviral compounds including ganciclovir, foscarnet, and cidofovir are routinely used in the treatment of cytomegalovirus (CMV) infection and disease; however, these agents have a poor oral bioavailability and have the inconvenience and expense of intravenous administration. AIM OF THE STUDY: To evaluate the safety and efficacy of oral valganciclovir (VGCV) for preemptive treatment of CMV reactivation in the setting of allogeneic hematopoietic stem cell transplantation (HSCT). PATIENTS AND METHODS: We treated 15 patients receiving allogeneic HSCT from related (n=9) or unrelated (n=6) donors. In all patients, either the donor, host, or both were CMV Ig G positive pretransplant. Indication for therapy was preemptive treatment of CMV infection defined as one or two consecutive positive tests of pp65 antigenemia assay or CMV-polymerase chain reaction (PCR). VGCV was administered orally in a dosage of 900 mg b.i.d. for 2 weeks, followed by 450 mg b.i.d. for 2 additional weeks. RESULTS: Patients developed a positive CMV-PCR after a median of 52 days (range 37-427) post HSCT and a positive pp65 antigenemia after a median time of 74 days (range 37-427) post HSCT. Preemptive treatment with VGCV was started a median time of 56 days (range 37-429) after transplant. In all, 11 patients (73%) completed the 28 days of therapy with VGCV. All patients showed a complete clearance of the virus. The median time to achieve a negative CMV-PCR was 6 days (range 4-18). A relapse of CMV infection after VGCV preemptive therapy occurred in 6 patients (40%). No patient developed early or late CMV disease. Six patients (40%) presented hematological toxicity including neutropenia and/or thrombocytopenia that required drug discontinuation in 4 cases. CONCLUSION: VGCV administered as preemptive therapy for CMV infection in patients receiving an allogeneic HSCT showed promise for treating this frequent complication. Prospective randomized studies in this setting are mandatory to yield more definitive results. PMID- 17461995 TI - Invasive adenoviral infections in T-cell-depleted allogeneic hematopoietic stem cell transplantation: high mortality in the era of cidofovir. AB - BACKGROUND: Adenovirus (ADV) infection occurs in 5-21% of allogeneic hematopoietic stem cell transplants (HSCT). Symptomatic enteritis and hemorrhagic cystitis may be encountered but are seldom fatal. In contrast, mortality rates of up to 75% are reported for adenoviral pneumonia or hepatitis. Cidofovir is currently being increasingly used for treatment of adenoviral infections after HSCT. The efficacy of cidofovir in patients with invasive adenoviral infection is not established. FINDINGS: We reviewed 687 adult and pediatric patients who received allogeneic HSCT at our institution from 1998 through June 2005. ADV was isolated from 64 (9.3%) patients. Eleven patients received cidofovir for invasive disease occurring at median 39 days (range 3-145) post HSCT. The median age was 40 (range 6-61) years. Seventy-three percent received a T-cell-depleted graft and 18% had grade 3-4 graft-versus-host disease (GVHD) of the gut. Three out of 3 (100%) patients with adenoviral pneumonia died. One patient with hepatitis, cholecysitis, and viremia cleared the infection after 3 months. Two out of 7 (28.6%) patients with hemorrhagic colitis or cystitis died of ADV (1 with extensive GVHD). CONCLUSION: Mortality rates of ADV pneumonitis after allogeneic HSCT remain high in the era of cidofovir. Clinical trials are needed to evaluate management strategies for this life-threatening infection. PMID- 17461996 TI - Infections associated with ventricular assist devices: epidemiology and effect on prognosis after transplantation. AB - BACKGROUND: Ventricular assist devices (VADs) can be used as a bridge to orthotopic heart transplantation (OHT) in people with severe congestive heart failure. Although they can be inserted for an indefinite time period (unlike balloon pumps), they do carry a substantial risk of infection. We studied the epidemiology, microbiology, and consequences of infection in patients with VADs who ultimately had cardiac transplantation. METHODS: Records of VAD-supported patients at our institution between January 1995 and January 2005 were identified by ICD-9 code. Infection was classified as driveline infection, pocket infection, mediastinitis, or VAD endocarditis in increasing severity of illness. RESULTS: Of 73 patients identified by ICD-9 code, 60 had charts available for review. Of these 60, 72% had a VAD infection: 13 had VAD endocarditis; 3, mediastinitis; 25, pocket infection; and 29, driveline infection. The only association of infection (43 patients, 72%) and demography or underlying disease was that of endocarditis with older age (median age 59 vs. 53 years; P=0.02) and diabetes mellitus (13 patients, 30%; risk ratio 3.4; P=0.01). The duration of VAD support was longer in infected patients (median 125 days) vs. uninfected ones (25 days). Median survival measured from the time of VAD placement (although also true from the time of transplantation) was shorter in patients with VAD endocarditis (120 days) and pocket infection (350 days) vs. no infection (>2400 days) with a significant P=0.017 for endocarditis. Four patients had infections after transplantation that were caused by the same organism as their VAD infection. The predominant pathogens in VAD infection were Staphylococcus and Enterococcus spp. CONCLUSION: VAD use as a bridge to cardiac transplantation is associated with a large number of device-related infections. Patients with infected VADs, on average, wait longer for transplantation than patients with uninfected VADs, and patients with VAD endocarditis have a shorter survival than patients with no VAD infection or simple driveline infection. PMID- 17461997 TI - Safety of aerosolized liposomal versus deoxycholate amphotericin B formulations for prevention of invasive fungal infections following lung transplantation: a retrospective study. AB - BACKGROUND: Nebulized amphotericin B deoxycholate (AmBd) has been used to prevent invasive pulmonary aspergillosis after lung transplantation. METHODS: In this retrospective study we compared the safety and tolerability of nebulized AmBd and nebulized liposomal amphotericin B (L-AmB) in 38 consecutive lung transplant recipients. Progress notes, medication administration records, microbiology, and pulmonary function reports were reviewed. Histologic sections from lung tissue were examined. Plasma amphotericin B levels were measured. RESULTS: A total of 1206 doses of AmBd and 1149 doses of L-AmB were administered. Eighteen patients received AmBd only, 11 received L-AmB only, and 9 received the medications sequentially. The total number of complaints vs. the number of doses administered was 1.0% for AmBd-treated patients and 1.2% for L-AmB-treated patients. No differences were observed between the treatment groups on lung biopsy specimens. Plasma amphotericin B levels were <0.2-0.9 microg/mL in AmBd-treated patients and <0.2 microg/mL in L-AmB-treated patients. CONCLUSIONS: In lung transplant recipients, both inhaled AmBd and L-AmB were safe and well tolerated over a large number of medication exposures. PMID- 17461998 TI - Dual infection with polyomavirus BK and acyclovir-resistant herpes simplex virus successfully treated with cidofovir in a bone marrow transplant recipient. AB - A hematopoietic stem cell transplant recipient developed a mucosal herpes simplex virus-1 (HSV-1) infection while under acyclovir (ACV) treatment (HSV was later shown to be resistant to ACV). Concomitantly, the patient presented a hemorrhagic cystitis (HC) due to polyomavirus BK, for which intravenous cidofovir (CDV) was prescribed. The patient benefited from the broad-spectrum anti-DNA virus activity of CDV, and not only the HC resolved without signs of nephrotoxicity but also the HSV-1 lesions disappeared. This is the first report describing the effect of CDV on 2 simultaneous and unrelated DNA viral infections in an immunosuppressed transplant recipient. In addition, we describe here that this HSV-1 isolate possesses a unique phenotype and genotype. PMID- 17461999 TI - Donor-transmitted toxoplasmosis in liver transplant recipients: a case report and literature review. AB - Transmission of toxoplasmosis via liver transplantation is extremely uncommon. Here we report the case of a 52-year-old male liver transplant recipient who on day 32 post transplant developed pneumonia followed by respiratory failure. Donor transmitted toxoplasmosis was confirmed as the etiology by both serologic and molecular testing. We also review all previously published cases of toxoplasmosis in the English-language adult liver transplant literature. PMID- 17462000 TI - Ethmoidal sinusitis caused by Exserohilum rostratum in a patient with malignant lymphoma after non-myeloablative allogeneic peripheral blood stem cell transplantation. AB - We describe a patient with aggressive lymphoma who contracted an ethmoidal sinus infection due to Exserohilum rostratum after non-myeloablative allogeneic peripheral blood stem cell transplantation. E. rostratum is an extremely rare causative pathogen of invasive fungal infection. Phylogenetic tree analysis of the D1/D2 domains within the LSU rDNA identified the molecular structure of isolates. We believe this is the first description of E. rostratum infection in a patient who underwent hematopoietic stem cell transplantation. PMID- 17462001 TI - The onset of rapidly progressive neurologic deterioration after a brief gastrointestinal illness in a renal allograft recipient. AB - Renal transplant recipients are at increased risk for life-threatening complications, most commonly infections. Because of their impaired cell-mediated immunity, these patients are particularly susceptible to organisms that rely on intracellular survival and spread, such as Listeria monocytogenes. Despite being a food-borne pathogen, L. monocytogenes is associated with significant morbidity and mortality. Here we report the case of a renal transplant recipient who developed rapidly progressive neurological symptoms after a brief gastrointestinal illness. Magnetic resonance imaging scan of the brain showed a large mass that was identified as an abscess due to L. monocytogenes. Timely aspiration and antibiotic treatment resulted in complete recovery, as opposed to worse outcomes in the available case reports. We further review the epidemiology, microbiology, clinical presentation, and therapeutic options for listerial brain abscess. PMID- 17462002 TI - Two cases of hepatic zygomycosis in allogeneic stem cell transplant recipients and review of literature. AB - Gastrointestinal zygomycosis is a rare condition with a high mortality rate. We present 2 fatal cases of hepatic zygomycosis following allogeneic hematopoietic stem cell transplantation and review the literature. PMID- 17462003 TI - Coccidioidomycosis masked by symptoms of end-stage liver disease in transplant candidates. AB - Coccidioidomycosis is a common fungal infection in the southwestern United States. Two liver transplant candidates contracted coccidioidomycosis while awaiting transplantation in an endemic area, one of whom had successful transplantation despite unrecognized active mycosis. Symptoms and signs mistakenly attributed to terminal liver disease may actually be caused by coccidioidomycosis. PMID- 17462004 TI - Successful treatment of disseminated fusariosis with posaconazole during neutropenia and subsequent allogeneic hematopoietic stem cell transplantation. AB - We report the case of a 16-year-old girl with acute myelogenous leukemia with disseminated fusariosis, who responded to salvage posaconazole therapy. She subsequently received additional cytotoxic chemotherapy and allogeneic hematopoietic stem cell transplantation with posaconazole continued as secondary prophylaxis. Despite intensive immunosuppressive therapy for graft-versus-host disease, no recrudescence of infection occurred. PMID- 17462005 TI - Outcomes of invasive zygomycosis infections in renal transplant recipients. AB - Zygomycosis is an infrequent and difficult-to-treat fungal infection that is found in patients with underlying immunocompromised states. The advent of the lipid amphotericin B products has allowed for treatment with higher doses of therapy and less systemic toxicity. We reviewed the outcomes of 6 renal transplant recipients diagnosed with biopsy-proven invasive zygomycosis who received amphotericin B lipid complex (ABLC) in doses greater than 5 mg/kg between 2000 and 2004. All 6 patients had baseline diabetes mellitus, were receiving immunosuppressive agents, and subsequently underwent concomitant surgery. Three of the 6 patients that survived had undergone significant surgical debridement, reduction of their immunosuppression to minimal prednisone, and received prolonged course of ABLC at 10 mg/kg/day. All survivors lost graft function during the course of their therapy. The 3 patients who died all had delays in diagnosis of their disease and subsequent surgical and appropriate medical therapy. Therefore, in renal transplant recipients the early diagnosis of invasive zygomycosis is imperative along with early therapy with surgical debridement, reduced immunosuppression, and the use of high doses of ABLC. PMID- 17462006 TI - Ex vivo monitoring of human cytomegalovirus-specific CD8+ T-cell responses using QuantiFERON-CMV. AB - We have developed a novel diagnostic technology to monitor the human cytomegalovirus (HCMV)-specific CD8+ T-cell responses that is based on the detection of secreted interferon-gamma (IFN-gamma) in the whole blood (referred to as QuantiFERON -CMV). Evaluation of QuantiFERON -CMV in healthy individuals revealed that this technology was at least as sensitive and with some HCMV epitopes more sensitive than the ELISPOT for detecting ex vivo IFN-gamma. Results from QuantiFERON -CMV assays showed 97% (36/37 individuals) agreement with the anti-HCMV serology test in healthy individuals. Furthermore, we also show that this technology can be used to assess HCMV-specific T-cell responses in transplant patients. This study shows that QuantiFERON -CMV is a simple, reproducible, and reliable test for the detection of IFN-gamma in response to HCMV CD8+ T-cell epitopes, and may be a valuable diagnostic test for the detection of HCMV infection and a useful clinical tool for monitoring the immune response in immunosuppressed patients during therapy. PMID- 17462007 TI - Lack of vision, loss of sight: consequences of mandatory HTLV screening in corneal transplantation. PMID- 17462008 TI - Split target specificity of ResT: a design for protein delivery, site selectivity and regulation of enzyme activity? AB - The ResT telomere resolvase is responsible for maintaining the hairpin telomeres that cap the linear chromosome and minichromosomes of Borrelia burgdorferi. This enzyme acts at the tandem telomere junctions present within circular dimers resulting from DNA replication. ResT mediates the transesterification steps of resolution using a constellation of active site residues similar to that found in tyrosine recombinases and type IB topoisomerases. By combining this reaction mechanism with a hairpin binding module in its N-terminal domain, ResT reduces a fused telomere dimer into two hairpin monomers. ResT displays a split DNA binding specificity, with the N- and C-terminal domains targeting distinct regions of the telomere. This bi-specificity in binding is likely to be important in protein delivery, substrate selection and regulation of enzyme activity. PMID- 17462009 TI - Telomere resolution by Borrelia burgdorferi ResT through the collaborative efforts of tethered DNA binding domains. AB - Borrelia burgdorferi, a causative agent of Lyme disease, has a highly unusual segmented genome composed of both circular molecules and linear DNA replicons terminated by covalently closed hairpin ends or telomeres. Replication intermediates of the linear molecules are processed into hairpin telomeres via the activity of ResT, a telomere resolvase. We report here the results of limited proteolysis and mass spectroscopy to identify two main structural domains in ResT, separated by a chymotrypsin cleavage site between residues 163 and 164 of the 449 amino acid protein. The two domains have been overexpressed and purified. DNA electrophoretic mobility shift assays revealed that the C-terminal domain (ResT(164-449)) displays sequence-specific DNA binding to the box 3,4,5 region of the telomere, while the N-terminal domain (ResT(1-163)) exhibits sequence independent DNA binding activity. Further analysis by DNase I footprinting supports a model for telomere resolution in which the hairpin binding module of the N-terminal domain is delivered to the box 1,2 region of the telomere through its tethering to ResT(164-449). Conversely, ResT(1-164) may play an important regulatory role by modulating both sequence-specific DNA binding activity and catalysis by the C-terminal domain. PMID- 17462010 TI - TUP1 disruption in Cryptococcus neoformans uncovers a peptide-mediated density dependent growth phenomenon that mimics quorum sensing. AB - Cryptococcus neoformans is a pathogenic yeast that causes life-threatening meningoencephalitis and grows well on mycological media regardless of inoculum size. Interestingly, a deletion of the global repressor TUP1 in C. neoformans uncovered a density-dependent growth phenotype reminiscent of the quorum-sensing phenomenon. An inoculum size of lower than 10(3) cells of the tup1Delta strain failed to form colonies on agar media while inocula of 10(5)-10(6) cells per plate formed a lawn. This phenotype, expressed as the inability to grow at low cell densities, was rescued by the culture filtrate from a high cell density tup1Delta culture and the active molecule in this culture filtrate was identified to be an oligopeptide composed of 11 amino acids. Activity assays, using a synthetic version of the peptide with strains harbouring a deletion of the corresponding gene, proved that the oligopeptide functioned as an autoregulatory molecule responsible for the density-dependent phenotype. Although a density dependent growth phenotype has been reported in several species of Ascomycetes, no peptide has been reported to function as an autoregulator in the Kingdom Fungi. The identification of an 11-mer peptide as an autoregulatory molecule in C. neoformans suggests that a diverse mechanism of cell-to-cell communication exists in the Kingdom Fungi. PMID- 17462011 TI - SapB and the chaplins: connections between morphogenetic proteins in Streptomyces coelicolor. AB - Morphogenesis in the streptomycetes features the differentiation of substrate associated vegetative hyphae into upwardly growing aerial filaments. This transition requires the activity of bld genes and the secretion of biosurfactants that reduce the surface tension at the colony-air interface enabling the emergence of nascent aerial hyphae. Streptomyces coelicolor produces two classes of surface-active molecules, SapB and the chaplins. While both molecules are important for aerial development, nothing is known about the functional redundancy or interaction of these surfactants apart from the observation that aerial hyphae formation can proceed via one of two pathways: a SapB-dependent pathway when cells are grown on rich medium and a SapB-independent pathway on poorly utilized carbon sources such as mannitol. We used mutant analysis to show that while the chaplins are important, but not required, for development on rich medium, they are essential for differentiation on MS (soy flour mannitol) medium, and the corresponding developmental defects could be suppressed by the presence of SapB. Furthermore, the chaplins are produced by conditional bld mutants during aerial hyphae formation when grown on the permissive medium, MS, suggesting that the previously uncharacterized SapB-independent pathway is chaplin dependent. In contrast, a bld mutant blocked in aerial morphogenesis on all media makes neither SapB nor chaplins. Finally, we show that a constructed null mutant that lacks all chaplin and SapB biosynthetic genes fails to differentiate in any growth condition. We propose that the biosurfactant activities of both SapB and the chaplins are essential for normal aerial hyphae formation on rich medium, while chaplin biosynthesis and secretion alone drives aerial morphogenesis on MS medium. PMID- 17462012 TI - Sre1p, a regulator of oxygen sensing and sterol homeostasis, is required for virulence in Cryptococcus neoformans. AB - Cryptococcus neoformans is an environmental pathogen requiring atmospheric levels of oxygen for optimal growth. Upon inhalation, C. neoformans disseminates to the brain and causes meningoencephalitis, but the mechanisms by which the pathogen adapts to the low-oxygen environment in the brain have not been investigated. We found that SRE1, a homologue of the mammalian sterol regulatory element-binding protein (SREBP), functions in an oxygen-sensing pathway. Low oxygen decreased sterol synthesis in C. neoformans and triggered activation of membrane-bound Sre1p by the cleavage-activating protein, Scp1p. Microarray and Northern blot analysis demonstrated that under low oxygen, Sre1p activates genes required for ergosterol biosynthesis and iron uptake. Consistent with these regulatory functions, sre1Delta cells were hypersensitive to azole drugs and failed to grow under iron-limiting conditions. Importantly, sre1Delta cells failed to produce fulminating brain infection in mice. Our in vitro data support a model in which Sre1p is activated under low oxygen leading to the upregulation of genes required for sterol biosynthesis and growth in a nutrient-limiting environment. Animal studies confirm the importance of SRE1 for C. neoformans to adapt to the host environment and to cause fatal meningoencephalitis, thereby identifying the SREBP pathway as a therapeutic target for cryptococcosis. PMID- 17462013 TI - Control of the respiratory metabolism of Thermus thermophilus by the nitrate respiration conjugative element NCE. AB - The strains of Thermus thermophilus that contain the nitrate respiration conjugative element (NCE) replace their aerobic respiratory chain by an anaerobic counterpart made of the Nrc-NADH dehydrogenase and the Nar-nitrate reductase in response to nitrate and oxygen depletion. This replacement depends on DnrS and DnrT, two homologues to sensory transcription factors encoded in a bicistronic operon by the NCE. DnrS is an oxygen-sensitive protein required in vivo to activate transcription on its own dnr promoter and on that of the nar operon, but not required for the expression of the nrc operon. In contrast, DnrT is required for the transcription of these three operons and also for the repression of nqo, the operon that encodes the major respiratory NADH dehydrogenase expressed during aerobic growth. Thermophilic in vitro assays revealed that low DnrT concentrations allows the recruitment of the T. thermophilus RNA polymerase sigma(A) holoenzyme to the nrc promoter and its transcription, whereas higher DnrT concentrations are required to repress transcription on the nqo promoter. In conclusion, our data show a complex autoinducible mechanism by which DnrT functions as the transcriptional switch that allows the NCE to take the control of the respiratory metabolism of its host during adaptation to anaerobic growth. PMID- 17462014 TI - ExoR is genetically coupled to the ExoS-ChvI two-component system and located in the periplasm of Sinorhizobium meliloti. AB - Sinorhizobium meliloti enters into a symbiotic relationship with legume host plants, providing fixed nitrogen in exchange for carbon and amino acids. In S. meliloti, exoR and the exoS-chvI two-component system regulate the biosynthesis of succinoglycan, an exopolysaccharide important for host invasion. It was previously reported that a loss-of-function mutation in exoR and a gain-of function mutation in exoS cause overproduction of succinoglycan and loss of motility, indicating that ExoR negatively regulates and ExoS-ChvI positively regulates downstream genes. However, a relationship between exoR and exoS-chvI has never been clearly established. By identification and detailed characterization of suppressor strains, we provide genetic evidence that exoR and exoS-chvI control many similar phenotypes. These include succinoglycan production, symbiosis, motility, and previously uncharacterized prototrophy and biofilm formation, all of which are co-ordinately restored by suppressors. We further demonstrate that ExoR is located in the periplasm, suggesting that it functions to regulate downstream genes in a novel manner. In pathogenic bacteria closely related to S. meliloti, exoS-chvI homologues are required for virulence and the regulation of cell envelope composition. Our data suggest that periplasmically localized ExoR and ExoS-ChvI function together in a unique and critical regulatory system associated with both free-living and symbiotic states of S. meliloti. PMID- 17462015 TI - The Pseudomonas putida Crc global regulator is an RNA binding protein that inhibits translation of the AlkS transcriptional regulator. AB - The Crc protein is a global regulator that controls the hierarchical assimilation of carbon sources in Pseudomonads by inhibiting expression of several catabolic pathways. Crc does not bind DNA and its mechanism of action has remained elusive. Among other genes, Crc inhibits expression of alkS, the transcriptional activator of the Pseudomonas putida OCT plasmid alkane degradation pathway. AlkS activates expression of its own gene. In the presence of saturating AlkS levels, translational fusions of alkS to the lacZ reporter gene were responsive to Crc, but transcriptional fusions were not. In translational fusions, the first 33 nt of alkS mRNA, which includes up to position +3 relative to the translation start site, were sufficient to confer an efficient response to Crc. In vitro, purified Crc could bind specifically to an alkS mRNA fragment spanning positions +1 to +43, comprising the translation initiation region. We have previously shown that Crc has little effect on the stability of alkS mRNA. We conclude that Crc modulates AlkS levels by binding to the translation initiation region of alkS mRNA, thereby inhibiting translation. Because AlkS is an unstable protein present in limiting amounts, reducing its levels leads to decreased expression of all genes in the pathway. PMID- 17462016 TI - The rotational dynamics of kinetoplast DNA replication. AB - Kinetoplast DNA (kDNA), from trypanosomatid mitochondria, is a network containing several thousand catenated minicircles that is condensed into a disk-shaped structure in vivo. kDNA synthesis involves release of individual minicircles from the network, replication of the free minicircles and reattachment of progeny at two sites on the network periphery approximately 180 degrees apart. In Crithidia fasciculata, rotation of the kDNA disk relative to the antipodal attachment sites results in distribution of progeny minicircles in a ring around the network periphery. In contrast, Trypanosoma brucei progeny minicircles accumulate on opposite ends of the kDNA disk, a pattern that did not suggest kinetoplast motion. Thus, there seemed to be two distinct replication mechanisms. Based on fluorescence microscopy of the kDNA network undergoing replication, we now report that the T. brucei kinetoplast does move relative to the antipodal sites. Whereas the C. fasciculata kinetoplast rotates, that from T. brucei oscillates. Kinetoplast motion of either type must facilitate orderly replication of this incredibly complex structure. PMID- 17462017 TI - The RecJ DNase strongly suppresses genomic integration of short but not long foreign DNA fragments by homology-facilitated illegitimate recombination during transformation of Acinetobacter baylyi. AB - Homology-facilitated illegitimate recombination (HFIR) promotes genomic integration of foreign DNA with a single segment homologous to the recipient genome by homologous recombination in the segment accompanied by illegitimate fusion of the heterologous sequence. During natural transformation of Acinetobacter baylyi HFIR occurs at about 0.01% of the frequency of fully homologous recombination. The role of the 5' single-strand-specific exonuclease RecJ in HFIR was investigated. Deletion of recJ increased HFIR frequency about 20 fold compared with wild type while homologous recombination was not affected. Illegitimate fusion sites were predominantly located within 360 nucleotides away from the homology whereas in wild type most fusion sites were distal (500-2500 nucleotides away). RecJ overproduction reduced the HFIR frequency to half compared with wild type, and transformants with short foreign DNA segments were diminished, leading to on average 866 foreign nucleotides integrated per event (682 in wild type, 115 in recJ). In recJ always the 3' ends of donor DNA were integrated at the homology whereas in wild type these were 3' or 5'. RecJ apparently suppresses HFIR by degrading 5' non-homologous DNA tails at the post synaptic stage. We propose that the RecJ activity level controls the HFIR frequency during transformation and the amount of foreign DNA integrated per event. PMID- 17462018 TI - Whole-genome analysis of the chromosome partitioning and sporulation protein Spo0J (ParB) reveals spreading and origin-distal sites on the Bacillus subtilis chromosome. AB - We investigated the genome-wide DNA binding of the chromosome partitioning and sporulation protein and ParB family member Spo0J in Bacillus subtilis using chromatin immunoprecipitation and DNA microarrays. We identified 10 parS loci to which Spo0J binds, two of which were unexpectedly distant (> 1 Mb) from the origin of replication. We used all 10 sites to refine the consensus sequence for parS. We found that Spo0J spreads along the DNA around each site. Binding was near maximal levels up to 1.6 kb away from parS, and significantly above background as far away as 18 kb. Deletion of soj (parA) had little or no effect on spreading. In contrast, the spo0J93 allele appeared to cause a significant decrease in spreading in vivo, without significantly affecting the DNA binding affinity in vitro. spo0J93 causes a phenotype similar to that of a spo0J null mutant and alters the region thought to be involved in interaction between Spo0J dimers. Our findings indicate that spreading is important for in vivo function of Spo0J. Gene expression in areas near parS sites was similar in wild type and a spo0J null mutant, indicating that binding and spreading of Spo0J on DNA does not normally silence transcription of nearby genes. PMID- 17462019 TI - A novel compartment, the 'subapical stem' of the aerial hyphae, is the location of a sigN-dependent, developmentally distinct transcription in Streptomyces coelicolor. AB - Streptomyces coelicolor has nine SigB-like RNA polymerase sigma factors, several of them implicated in morphological differentiation and/or responses to different stresses. One of the nine, SigN, is the focus of this article. A constructed sigN null mutant was delayed in development and exhibited a bald phenotype when grown on minimal medium containing glucose as carbon source. One of two distinct sigN promoters, sigNP1, was active only during growth on solid medium, when its activation coincided with aerial hyphae formation. Transcription from sigNP1 was readily detected in several whi mutants (interrupted in morphogenesis of aerial mycelium into spores), but was absent from all bld mutants tested, suggesting that sigNP1 activity was restricted to the aerial hyphae. It also depended on sigN, thus sigN was autoregulated. Mutational and transcription studies revealed no functional significance to the location of sigN next to sigF, encoding another SigB-like sigma factor. We identified another potential SigN target, nepA, encoding a putative small secreted protein. Transcription of nepA originated from a single, aerial hyphae-specific and sigN-dependent promoter. While in vitro run off transcription using purified SigN on the Bacillus subtilis ctc promoter confirmed that SigN is an RNA polymerase sigma factor, SigN failed to initiate transcription from sigNP1 and from the nepA promoter in vitro. Additional in vivo data indicated that further nepA upstream sequences, which are likely to bind a potential activator, are required for successful transcription. Using a nepA-egfp transcriptional fusion we located nepA transcription to a novel compartment, the 'subapical stem' of the aerial hyphae. We suggest that this newly recognized compartment defines an interface between the aerial and vegetative parts of the Streptomyces colony and might also be involved in communication between these two compartments. PMID- 17462020 TI - An antisense RNA controls synthesis of an SOS-induced toxin evolved from an antitoxin. AB - Only few small, regulatory RNAs encoded opposite another gene have been identified in bacteria. Here, we report the characterization of a locus where a small RNA (SymR) is encoded in cis to an SOS-induced gene whose product shows homology to the antitoxin MazE (SymE). Synthesis of the SymE protein is tightly repressed at multiple levels by the LexA repressor, the SymR RNA and the Lon protease. SymE co-purifies with ribosomes and overproduction of the protein leads to cell growth inhibition, decreased protein synthesis and increased RNA degradation. These properties are shared with several RNA endonuclease toxins of the toxin-antitoxin modules, and we show that the SymE protein represents evolution of a toxin from the AbrB fold, whose representatives are typically antitoxins. We suggest that SymE promotion of RNA cleavage may be important for the recycling of RNAs damaged under SOS-inducing conditions. PMID- 17462021 TI - Molecular analysis of the cercosporin biosynthetic gene cluster in Cercospora nicotianae. AB - We describe a core gene cluster, comprised of eight genes (designated CTB1-8), and associated with cercosporin toxin production in Cercospora nicotianae. Sequence analysis identified 10 putative open reading frames (ORFs) flanking the previously characterized CTB1 and CTB3 genes that encode, respectively, the polyketide synthase and a dual methyltransferase/monooxygenase required for cercosporin production. Expression of eight of the genes was co-ordinately induced under cercosporin-producing conditions and was regulated by the Zn(II)Cys(6) transcriptional activator, CTB8. Expression of the genes, affected by nitrogen and carbon sources and pH, was also controlled by another transcription activator, CRG1, previously shown to regulate cercosporin production and resistance. Disruption of the CTB2 gene encoding a methyltransferase or the CTB8 gene yielded mutants that were completely defective in cercosporin production and inhibitory expression of the other CTB cluster genes. Similar 'feedback' transcriptional inhibition was observed when the CTB1, or CTB3 but not CTB4 gene was inactivated. Expression of four ORFs located on the two distal ends of the cluster did not correlate with cercosporin biosynthesis and did not show regulation by CTB8, suggesting that the biosynthetic cluster was limited to CTB1-8. A biosynthetic pathway and a regulatory network leading to cercosporin formation are proposed. PMID- 17462022 TI - Systematic capsule gene disruption reveals the central role of galactose metabolism on Cryptococcus neoformans virulence. AB - The polysaccharidic capsule is the main virulence factor of Cryptococcus neoformans. It primarily comprised of two polysaccharides: glucuronoxylomannan (GXM, 88% of the capsule mass) and galactoxylomannan (GalXM, 7% of the capsule mass). We constructed a large collection of mutant strains in which genes potentially involved in capsule biosynthesis were deleted. We used a new post genomic approach to study the virulence of the strains. Primers specific for unique tags associated with the disruption cassette were used in a real-time PCR virulence assay to measure the fungal burden of each strain in different organs of mice in multi-infection experiments. With this very sensitive assay, we identified a putative UDP-glucose epimerase (Uge1p) and a putative UDP-galactose transporter (Ugt1p) essential for C. neoformans virulence. The uge1Delta and ugt1Delta strains are temperature sensitive and do not produce GalXM but synthesize a larger capsule. These mutant strains (GalXM negative, GXM positive) are not able to colonize the brain even at the first day of infection whereas GXM negative strains (GalXM positive) can still colonize the brain, although less efficiently than the wild-type strain. PMID- 17462023 TI - The Anabaena sp. strain PCC 7120 asr1734 gene encodes a negative regulator of heterocyst development. AB - The novel asr1734 gene of Anabaena (Nostoc) sp. strain PCC 7120 inhibited heterocyst development when present in extra copies. Overexpression of asr1734 inhibited heterocyst development in several strains including the wild type and two strains that form multiple contiguous heterocysts (Mch phenotype): a PatS null mutant and a hetR(R223W) mutant. Overexpression of asr1734 also caused increased nblA messenger RNA levels, and increased loss of autofluorescence in vegetative cells throughout filaments after nitrogen or sulphur depletion. Unlike the wild type, an asr1734 knockout mutant formed 5% heterocysts after a nitrogen shift from ammonium to nitrate, and formed 15% heterocysts and a weak Mch phenotype after step-down to medium lacking combined nitrogen. After nitrogen step-down, the asr1734 mutant had elevated levels of ntcA messenger RNA. A green fluorescent protein reporter driven by the asr1734 promoter, P(asr1734)-gfp, was expressed specifically in differentiating proheterocysts and heterocysts after nitrogen step-down. Strains overexpressing asr1734 and containing P(hetR)-gfp or P(patS)-gfp reporters failed to show normal patterned upregulation 24 h after nitrogen step-down even though hetR expression was upregulated at 6 h. Apparent orthologues of asr1734 are found only in two other filamentous nitrogen-fixing cyanobacteria, Anabaena variabilis and Nostoc punctiforme. PMID- 17462024 TI - Identification of a system required for the functional surface localization of sugar binding proteins with class III signal peptides in Sulfolobus solfataricus. AB - The hyperthermophilic archaeon Sulfolobus solfataricus contains an unusual large number of sugar binding proteins that are synthesized as precursors with a class III signal peptide. Such signal peptides are commonly used to direct archaeal flagellin subunits or bacterial (pseudo)pilins into extracellular macromolecular surface appendages. Likewise, S. solfataricus binding proteins have been suggested to assemble in higher ordered surface structures as well, tentatively termed the bindosome. Here we show that S. solfataricus contains a specific system that is needed for the functional surface localization of sugar binding proteins. This system, encoded by the bas (bindosome assembly system) operon, is composed of five proteins: basABC, three homologues of so-called bacterial (pseudo)pilins; BasE, a cytoplasmic ATPase; and BasF, an integral membrane protein. Deletion of either the three (pseudo)pilin genes or the basEF genes resulted in a severe defect of the cells to grow on substrates which are transported by sugar binding proteins containing class III signal peptides, while growth on glucose and maltose was restored when the corresponding genes were reintroduced in these cells. Concomitantly, DeltabasABC and DeltabasEF cells were severely impaired in glucose uptake even though the sugar binding proteins were normally secreted across the cytoplasmic membrane. These data underline the hypothesis that the bas operon is involved in the functional localization of sugar binding proteins at the cell surface of S. solfataricus. In contrast to surface structure assembly systems of Gram-negative bacteria, the bas operon seems to resemble an ancestral simplified form of these machineries. PMID- 17462025 TI - Cold-shock-induced de novo transcription and translation of infA and role of IF1 during cold adaptation. AB - Escherichia coli infA is transcribed from two promoters, P1 and P2, into a longer and a shorter mRNA encoding translation initiation factor IF1. Although P1 is intrinsically stronger than P2, the shorter half-life of its transcripts causes the steady-state level of the P2 transcript to be substantially higher than that of P1 during growth at 37 degrees C. After cold-shock, de novo transcription and translation of infA contribute to the transient increase of the IF1/ribosomes ratio, which is partially responsible for translational bias consisting in the preferential translation of cold-shock mRNAs in the cold. Cold-stress induction of infA expression is mainly due to the high activity of P1 at low temperature, which is further increased by transcriptional stimulation by CspA and by an increased transcript stability. Furthermore, the longer infA mRNA originating from P1 is preferentially translated at low temperature by the translational machinery of cold-shocked cells. The increased level of IF1 during cold adaptation is essential for overcoming the higher stability of the 70S monomers at low temperature and for providing a sufficient pool of dissociated 30S subunits capable of initiating translation. PMID- 17462026 TI - Submicromolar hydrogen peroxide disrupts the ability of Fur protein to control free-iron levels in Escherichia coli. AB - In aerobic environments, mutants of Escherichia coli that lack peroxidase and catalase activities (Hpx(-)) accumulate submicromolar concentrations of intracellular H(2)O(2). We observed that in defined medium these strains constitutively expressed members of the Fur regulon. Iron-import proteins, which Fur normally represses, were fully induced. H(2)O(2) may antagonize Fur function by oxidizing the Fur:Fe(2+) complex and inactivating its repressor function. This is a potential problem, as in iron-rich environments excessive iron uptake would endanger H(2)O(2)-stressed cells by accelerating hydroxyl-radical production through the Fenton reaction. However, the OxyR H(2)O(2)-response system restored Fur repression in iron-replete Luria-Bertani medium by upregulating the synthesis of Fur protein. Indeed, when the OxyR binding site upstream of fur was disrupted, Hpx(-) mutants failed to repress transporter synthesis, and they exhibited high levels of intracellular free iron. Mutagenesis and bacteriostasis resulted. These defects were eliminated by mutations or chelators that slowed iron import, confirming that dysregulation of iron uptake was the root problem. Thus, aerobic organisms must grapple with a conundrum: how to monitor iron levels in oxidizing environments that might perturb the valence of the analyte. The induction of Fur synthesis by the OxyR response comprises one evolutionary solution to that problem. PMID- 17462027 TI - Cwp2p, the plasma membrane receptor for Pichia membranifaciens killer toxin. AB - PMKT is a channel-forming killer toxin secreted by Pichia membranifaciens. To identify novel genes that mediate cellular resistance to PMKT we screened a collection of 288 deletion mutants. We found 29 open reading frames (ORFs) that, when deleted, confer resistance to PMKT. In addition, the deletion of 15 ORFs was observed to increase protoplast resistance, in agreement with the initial assumption that a plasma membrane receptor for PMKT exists. Whole cells and protoplasts of a cwp2Delta mutant were found to be completely resistant to PMKT and were unable to bind PMKT, indicating that Cwp2p interacts with it. A protein with a molecular mass of 11.7 kDa was purified from PMKT-affinity columns. This protein was sequenced and identified as Cwp2p. Glycosylphosphatidylinositol (GPI) anchoring-defective mutants were much less sensitive to PMKT, as were wild-type protoplasts pretreated with phosphatidylinositol-specific phospholipase C to remove GPI-anchored proteins, indicating that the GPI-anchored precursor of Cwp2p is also necessary for PMKT activity. Carboxyfluorescein-entrapped liposomes containing a purified GFP-Cwp2p fusion protein in their membranes were much more sensitive to PMKT than protein-free liposomes. Cwp2p and its GPI-anchored precursor are proposed for the first time to be involved as PMKT secondary receptors. PMID- 17462028 TI - Mechanism of cell surface expression of the Streptococcus mitis platelet binding proteins PblA and PblB. AB - PblA and PblB are prophage-encoded proteins of Streptococcus mitis strain SF100 that mediate binding to human platelets. The mechanism for surface expression of these proteins has been unknown, as they do not contain signal sequences or cell wall sorting motifs. We therefore assessed whether expression of these proteins was linked the lytic cycle of the prophage. Deletion of either the holin or lysin gene resulted in retention of PblA and PblB in the cytoplasm, and loss of these proteins from the cell wall. Flow cytometric analysis revealed that induction of phage replication in SF100 produced a subpopulation of cells with increased permeability. This effect was abrogated by disruption of the holin and lysin genes. Treatment of these mutants with exogenous PblA and PblB restored surface expression, apparently via binding of the proteins to cell wall choline. Loss of PblA and PblB expression was associated with decreased platelet binding in vitro, and reduced virulence in an animal model of endocarditis. Thus, expression of PblA and PblB occurs via a novel mechanism, whereby phage induction increases bacterial permeability and release of the proteins, followed by their binding to surface of viable cells. This mechanism may be important for endovascular infection. PMID- 17462029 TI - Protein mobility and diffusive barriers in Escherichia coli: consequences of osmotic stress. AB - The effect of osmotic stress on the intracellular diffusion of proteins in Escherichia coli was studied, using a pulsed version of fluorescence recovery after photo-bleaching, pulsed-FRAP. This method employs sequences of laser pulses which only partly bleach the fluorophores in a cell. Because the cell size and geometry are taken into account, pulsed-FRAP enables to measure diffusion in very small cells of different shapes. We found that upon an osmotic upshock from 0.15 to 0.6 Osm, imposed by NaCl or sorbitol, the apparent intracellular diffusion (D) of mobile green fluorescent protein (GFP) decreased from 3.2 to 0.4 microm(2) s( 1), whereas the membrane permeable glycerol had no effect. Exposing E. coli cells to higher osmolalities (> 0.6 Osm) led to compartmentalization of the GFP into discrete pools, from where the GFP could not escape. Although free diffusion through the cell was hindered, the mobility of GFP in these pools was still relatively high (D approximately 0.4 microm(2) s(-1)). The presence of osmoprotectants restored the effect of osmotic stress on the protein mobility and apparent compartmentalization. Also, lowering the osmolality from 0.6 Osm back to 0.15 Osm restored the mobility of GFP. The implications of these findings in terms of heterogeneities and diffusive barriers inside the cell are discussed. PMID- 17462032 TI - Patients' experiences of being delirious. AB - AIM: The aim was to describe patients' experiences of being delirious. BACKGROUND: Delirium is a serious psychiatric disorder that is frequently reported from hospital care settings, particularly among older patients undergoing hip surgery. It involves disturbances of consciousness and changes in cognition, a state which develops over a short period of time and tends to fluctuate during the course of the day. It is a certified fact that delirium is poorly diagnosed and recognized although the state often is described as terrifying. To be able to give professional care, it is of the utmost importance to know more about patients' experience of delirium. METHOD: Included in the interviews were patients who had undergone hip-related surgery and during the hospital stay experienced delirium. Fifteen patients participated in the interviews. Of these, six had experienced episodes of nightly delirium (sundown syndrome) and nine experienced delirium during at least one day. The interviews were analysed by qualitative content analysis. RESULTS: The entry of delirium was experienced as a sudden change of reality that, in some cases, could be connected to basic unfulfilled physiological needs. The delirium experiences were like dramatic scenes that gave rise to strong emotional feelings of fear, panic and anger. The experiences were also characterized by opposite pairs; they took place in the hospital but at the same time somewhere else; it was like dreaming but still being awake. The exit from the delirium was associated with disparate feelings. RELEVANCE TO CLINICAL PRACTICE: It is necessary to understand patients' thoughts and experiences during the delirious phase to be able to give professional care, both during the delirium phase and after the recovery. PMID- 17462033 TI - Factors associated with delirium severity among older patients. AB - AIM: The goal of this study was to determine whether the factors associated with delirium varied according to the severity of the delirium experienced by the older patients. BACKGROUND: Delirium among older patients is prevalent and leads to numerous detrimental effects. The negative consequences of delirium are worse among older adults with severe delirium compared with patients with mild delirium. There has been no study identifying those factors associated with delirium severity among long-term care older patients newly admitted to an acute care hospital. DESIGN: This is a descriptive study. METHODS: This is a secondary analysis study of institutionalized older patients newly admitted to an acute care hospital (n = 104). Upon admission, patients were screened for delirium with the Confusion Assessment Method and severity of delirium symptoms were determined by using the Delirium Index. RESULTS: Of the 71 delirious older patients, 32 (45.1%) had moderate-severe delirium while 39 (54.9%) presented mild delirium. In univariate analyses, a significant positive relationship was observed between the level of prior cognitive impairment and the severity of delirium (p = 0.0058). Low mini-mental state examination (MMSE) scores (p < 0.0001), the presence of severe illness at the time of hospitalization (p = 0.0016) and low functional autonomy (BI: p = 0.0017; instrumental activities of daily living: p = 0.0003) were significantly associated with moderate-severe delirium. Older patients suffering from mild delirium used significantly more drugs (p = 0.0056), notably narcotics (p = 0.0017), than those with moderate-severe delirium. Results from the stepwise regression indicated that MMSE score at admission and narcotic medication use are the factors most strongly associated with the severity of delirium symptoms. CONCLUSIONS: This present study indicates that factors associated with moderate-severe delirium are different from those associated with mild delirium. Given the result concerning the role of narcotics, future studies should evaluate the role of pain management in the context of delirium severity. RELEVANCE TO CLINICAL PRACTICE: As moderate-severe delirium is associated with poorer outcomes than is mild delirium, early risk factor identification for moderate-severe delirium by nurses may prove to be of value in preventing further deterioration of those older patients afflicted with delirium. PMID- 17462034 TI - The association between caregiving satisfaction, difficulties and coping among older family caregivers. AB - AIMS AND OBJECTIVES: The aim was to study the association between gender, extent and content of care, satisfaction, coping and difficulties in the caregiving situation among older (75+) caregivers and to identify clusters of caregivers. The aim was also to explore psychometrically two instruments assessing satisfaction and difficulties in family caregivers. BACKGROUND: Caregiving is a complicated phenomenon. Much of the research has focused on negative aspects, such as the burden, stress and emotional strain. Caregiving is known to affect health negatively for the caregivers. Little is known about satisfaction and motivation in voluntary work, such as informal caregiving, especially among older persons. DESIGN AND METHODS: Cross-sectional. The sample for this study consisted of 171 informal caregivers aged 75 and over, identified from an age-stratified sample in a postal survey among older people in the southern part of Sweden. RESULTS: Male caregivers proved to be more satisfied than female caregivers; caregiving had seemingly widened their horizon and had helped them to grow as persons. Based on satisfaction scores, those satisfied had a higher proportion of male caregivers and a significantly higher amount of caregiving hours per week. They used other coping strategies than the respondents in the other cluster, i.e. less satisfied in using more problem-solving strategies. CONCLUSIONS: The instruments tested were appropriate for work in clinical and research settings, although the internal dropout indicates that a shorter version would be more useful. Those who found satisfaction in care used more problem-focused coping strategies and were more often men than women. From a salutogenic point of view, this may give important knowledge about factors that can promote health. The findings indicate that women deserve extra attention as informal caregivers as they did not find caregiving as rewarding as the men did. This may in turn make them less protected against the negative consequences of caregiving. RELEVANCE TO CLINICAL PRACTICE: Reinforcing the health-promoting qualities in caregivers who are not feeling well, with women as a particularly vulnerable group, may restrict unnecessary suffering for both the caregiver and the person cared for. PMID- 17462035 TI - Admitted with a hip fracture: patient perceptions of rehabilitation. AB - AIMS AND OBJECTIVES: The aim of this study was to describe the hip fracture patients' own perceptions of their situation and views of their responsibility in the rehabilitation process. BACKGROUND: Although much research has been conducted on various aspects of the rehabilitation process in patients with a hip fracture, no attention has been given to the patients' own views of their situation at the start of this transitional process. METHOD: Thirteen informants with a hip fracture, aged 71-93 years, were interviewed postoperatively at a Swedish hospital. Phenomenographic analysis of the interview transcripts was performed. RESULTS: The informants varied greatly in their engagement in the rehabilitation process, in their conceptions of who was responsible for their recovery and in their views on the need for information pertinent to their condition. Three categories of description were formulated: the Autonomous, i.e. patients who were self-sufficient and used to taking care of themselves and who searched for relevant information; the Modest, i.e. frail patients in need of more support who wanted information, but did not ask for it; and the Heedless, i.e. patients who were already dependent, who were not aware of their own responsibility and not interested in information. The informants also shared some traits: they all needed more information although not all were aware of it, they all worried about their future ability to walk again and they all had a strong zest for life. RELEVANCE TO CLINICAL PRACTICE: Our results suggest that differences in patients' perspectives on the rehabilitation process need to be taken into account to enhance outcomes. Inadequate knowledge and engagement on the part of patients, with a hip fracture, probably have an impact on their rehabilitation outcome, but the degree of impact is uncertain. PMID- 17462036 TI - Aged care nursing in Queensland - the nurses' view. AB - AIMS AND OBJECTIVES: Through comparison of two studies undertaken three years apart the opinions of nurses working in aged care facilities in Queensland were determined. Results will support policy planning for the Queensland Nurses Union. BACKGROUND: An ageing population in Australia is placing increased demands on residential aged care facilities. In Queensland, the national situation is exacerbated by an influx of retirees from other states and territories. The ongoing problem of shortages of nurses in the workforce may be addressed by gaining further insight into the nurses' own views of their conditions and experiences. METHODS: One thousand nurses working in public and privately owned residential aged care facilities were surveyed by postal questionnaire in 2004. Results were compared with data collected in an identical study in 2001. RESULTS: Respondents offered their opinions on working hours and conditions, professional development and experiences in nursing. The predominately female aged care nursing workforce is ageing. Reported workplace violence has increased substantially since 2001. Some improvements are reported in staff numbers, skill mix and workplace policies. Nurses expressed very serious concerns about pay, workload, stress, physical and emotional demands and staff morale. CONCLUSION: Working conditions for nurses in the residential aged care sector in Queensland must be addressed to retain the current nurses and to encourage new nurses to replace those that retire. RELEVANCE TO CLINICAL PRACTICE: The findings of this study provide information not only for the Queensland Nurses Union but also policy makers and nurse managers both nationally and internationally on areas that need to be addressed to maintain the required workforce within the aged care sector. PMID- 17462037 TI - Nurses' perceptions of quality and the factors that affect quality care for older people living in long-term care settings in Ireland. AB - AIM: The aim of this research was to determine the factors that facilitate or hinder high quality nursing care for older people in long-term care settings in Ireland. BACKGROUND: The quality of care for older people living in long-term care has been identified as an issue of concern in many nursing research studies. While many factors which have an impact on care have been identified, it is difficult to determine key factors from current research. METHOD: This was a mixed method study, which involved qualitative and quantitative approaches. A self-response questionnaire was generated from data gathered by interview and analysis of literature. Information was collected from 498 nurses working in long term care settings within a Health Board. A 68% response rate was achieved. Factor analysis was used to identify facilitating and hindering factors of quality care for older people. RESULTS: Nine factors where identified six facilitating factors of quality and three hindering factors of quality care. The six factors, which facilitate quality, were: an ethos of promoting independence and autonomy; a homelike social environment; person centred, holistic care; knowledgeable, skilled staff; knowing the person and adequate multidisciplinary resources. The three factors which hindered quality care; these were: a lack of time and patient choice, resistance to change and bound by routine. CONCLUSIONS: The findings of this research provide nurses with a clear set of facilitating and hindering factors of quality care for older people and reveal some of the complexities and challenges of providing this care. RELEVANCE TO CLINICAL PRACTICE: Long-term care is the home of many older people and attention within these environments to the facilitating and hindering factors of quality is required. It is hoped that the factors generated in this study add to understanding in relation to quality care and the factors that influence this. PMID- 17462038 TI - Predictors of psychological health in spouses of persons affected by stroke. AB - AIM: To identify predictors of psychological health and examine if these predictors change over time in spouses of stroke patients during the first year after stroke. A second aim was to identify gender differences in psychological health among the spouses. BACKGROUND: The impact of burden in long-term caregivers may result in psychological consequences for the spouse. The rehabilitation process for the patient can be negatively affected by a stressed caregiver and result in long-term hospitalization. To identify spouses at risk for physical and psychological distress is, therefore, essential to support those in need. DESIGN: Longitudinal, comparative study. METHODS: One hundred spouses of stroke patients were assessed at baseline, as well as after six and 12 months, regarding psychological health, well-being, own illness, need of assistance from general practitioner and/or district nurse, social network and knowledge about stroke. Stepwise multiple regression analyses were conducted for baseline, six- and 12-month assessments, respectively, with psychological health as the dependent variable. RESULTS: General well-being and presence of illness in spouse were the most prominent predictors of psychological health, throughout the first year. CONCLUSIONS: Enhancing psychological health and preventing medical problems in the caregiver are essential considerations to enable patients with stroke related disabilities to continue to live at home. RELEVANCE TO CLINICAL PRACTICE: Evaluating the situation for spouses of stroke patients is an important component when planning for the future care of the patient. PMID- 17462039 TI - Patients experiencing local anaesthesia and hip surgery. AB - AIMS AND OBJECTIVES: The aim of this study was to show what the experience of local anaesthesia and a surgical situation meant to patients. BACKGROUND: Many patients who receive local anaesthesia will remain awake during surgery. The very fact of staying awake causes psychological and psychosocial needs. These needs must be met and it is therefore important to identify them. DESIGN/METHOD: Seven patients (aged 61-79) experiencing local anaesthesia and hip surgery (hip replacement surgery or repair of fractured hip) were interviewed. An interpretive phenomenological method developed by Benner was used to extract the experience of what it means to be in local anaesthesia and surgery. One paradigm case is used to illustrate the results. RESULTS: Results show that the well-being and comfort of patients is compromised by challenges such as severe pain and long waits, which may be experienced as endless and which leave the patient thinking of nothing else. By contrast, the experience of trust helps the patient to feel control even in situations where the treatment is hard to grasp. CONCLUSIONS: The results highlight that local anaesthesia and surgery force patients to overcome and handle experiences of pain, trust and distrust as well as feelings of alienation and unreality. RELEVANCE TO CLINICAL PRACTICE: The results illustrate the patient's needs, as seen from the patient's perspective. A shared experience makes it possible to understand, and thus to recognize, the required qualitative care to facilitate and to help the patient to remain in control and face the psychological challenges presented by anaesthesia care and surgical situations. PMID- 17462040 TI - Violence in municipal care of older people in Sweden as perceived by registered nurses. AB - AIMS: The main aim was to describe registered nurses' perceptions of violence and threats, as well as their access to prevention measures and routines for handling violent behaviour in municipal care of older people. Another aim was to compare nurses' perceptions working solely in dementia care with those working in general elder care where older people have diverse diagnoses. BACKGROUND: Violence is often reported in care of older people. The development of dementia units and the implementation of reform have changed care of older people. Dementia disorders have been shown to be a predisposing factor to violence. DESIGN: A non experimental, descriptive design with a survey research approach was used. The setting was 60 special types of housing with subunits in a large town. The response rate was 62% (n = 213). Forty-five per cent (n = 95) of the nurses worked in dementia care and 55% (118) in general elder care. METHOD: A questionnaire. Results. Nurses had experienced a high degree indirect threats (dementia care, 45%; general elder care, 51%), direct threats of violent acts (dementia care, 35%; general elder care, 44%) and violent acts (dementia care, 41%; general elder care, 43%). Nurses had witnessed violence and threats towards staff (dementia care, 49%; general elder care, 38%). Even care receivers (dementia care, 20%; general elder care, 19%) were subjected to violence and threats. No statistical differences were found between groups. The nurses in dementia care had more access to education in managing violence and threats, as well as routines for handling violence and a door with a lock to their working unit. CONCLUSION: Violence occurred frequently in municipal care of older people without any difference between dementia care and general elder care. Nurses in dementia care were more often offered education on how to manage violence and had routines for when violence occurs. RELEVANCE TO CLINICAL PRACTICE: Municipal authorities should increase staff education for handling violence and creating safety routines. Violence needs to be taken seriously with a 'zero tolerance' attitude. PMID- 17462041 TI - Nursing home placement: perspectives of community-dwelling older persons. AB - OBJECTIVES: To explore nursing home-related beliefs among older Hong Kong Chinese and identify strategies that make nursing homes a better place to live. BACKGROUND: Increased life expectancy has markedly increased the prevalence of age-related disease and disability, which limits the self-caring ability of older persons. Nursing home placement offers one possible alternative. However, the perspectives of older persons on nursing home admission have been seriously neglected and under-reported. METHODS: The research was a qualitative descriptive study. Clients of a community centre for older people were approached and invited for an interview. RESULTS: Participants included 118 older persons (61 female and 57 male) age from 60-89 years. Ninety per cent had chronic diseases and needed regular follow-up. Half of participants had never visited a nursing home. Of them, 85% would not want to live in a nursing home upon getting old and their nursing home-related beliefs were found to be negative. Participants expressed the feeling of being abandoned by their family members, insecure and lonely regarding the nursing home placement. Living in a nursing home was considered as 'living in jail' and that their privacy would be lost. Nevertheless, positive aspects of nursing home placement were mentioned, and strategies were explored that would make nursing homes a better place for older people to live. RELEVANCE TO CLINICAL PRACTICE: Positive aspects of nursing home placement need to be promoted. Nurses should be advocates for the health of the elderly, and help make nursing homes a better place to live. PMID- 17462042 TI - Maintaining the integrity of older patients in long-term institutions: relatives' perceptions. AB - AIM: To study relatives' views on how the integrity of older patients is maintained in long-term institutions in Finland. BACKGROUND: aintaining patient integrity is considered a key prerequisite for good quality care and treatment and a characteristic of ethical care. Institutionalization, dependence on others and vulnerability may all threaten the integrity of the older patient and, in this sense, pose ethical problems. DESIGN/METHODS: The data for this descriptive and explorative survey were collected by questionnaires from 213 relatives of older patients in four long-term institutions in Finland. The response rate was 78%. Data were analysed using statistical methods. RESULTS: Relatives rated the maintenance of patient integrity quite highly. The highest ratings were recorded for the maintenance of physical integrity and the lowest to the maintenance of psychological integrity. The main source of problems were patients' and/or relatives' wishes concerning elimination. There were also shortcomings in the provision of intimacy. Relatives who thought that admission to the long-term institution had been problematic, either for the patient or for themselves, felt that patient integrity was poorly maintained. CONCLUSION: The findings highlight the importance of nurses' efforts in long-term care facilities to preserve the integrity of patients and to maintain close contact and cooperation with relatives. Although relatives generally were quite pleased with how patient integrity was maintained, the results also provide evidence on problems that may threaten patient integrity. RELEVANCE TO CLINICAL PRACTICE: Greater emphasis should be placed on the need for cooperation between older patients, their relatives and nursing staff, especially before admission to long-term care. Furthermore, increased efforts are needed in maintaining the integrity of older patients and in reducing ethically problematic situations. PMID- 17462043 TI - Effectiveness of bran supplement in older orthopaedic patients with constipation. AB - AIM: The purpose of the study was to evaluate the effectiveness of planned nursing interventions, including bran supplement, on the bowel management of older orthopaedic patients. BACKGROUND: Constipation is prevalent among older people. Constipation is also well-known to be a problem for a range of orthopaedic patients. Orthopaedic diseases or conditions may, because of the specific problems, cause this. The problem may also arise as a result of the orthopaedic treatment options undertaken by patients. DESIGN: A quasiexperimental design was employed. METHODS: The patients were recruited from an orthopaedic clinic at a university hospital in Erzurum, eastern Turkey. Descriptive statistics, independent sample test t-test, chi-square and McNemar test were used to analyse the data. Constipation problems were identified through interviews by using the Constipation Diagnosis Form. Interviews were performed within the fourth postoperative day. After that, while the patients in the control group received routine nursing care according to clinic routine, the patients in the experimental group received bran supplement together with planned nursing interventions. The patents in both groups were followed by using the Constipation Follow Form. RESULTS: While patients in the experimental group showed significant improvements in most characteristics of bowel elimination such as time of defecation, intensity of faeces, colour of faeces and amount of faeces, the control group showed a significant improvement in only duration of defecation. CONCLUSIONS: The results of the study indicate planned nursing interventions including bran supplement are more effective than routine nursing interventions for management of constipation problems in older orthopaedic patients. RELEVANCE TO CLINICAL PRACTICE: Nurses have an important part to play in both prevention and management of constipation. Constipation is a problem especially for older orthopaedic patients. Planned nursing interventions that include bran supplements may be more effective than routine nursing interventions for management of constipation problems in older orthopaedic patients. PMID- 17462044 TI - Betwixt and between: a critical ethnography of comfort in New Zealand residential aged care. AB - AIMS AND OBJECTIVES: This research sought to explore the nature of comfort within the context of three New Zealand nursing homes and examine how nursing and other actions contribute to residents' comfort. BACKGROUND: Comfort has been acknowledged as a key nursing goal since the days of Florence Nightingale, but the nature of comfort and how nurses can contribute to and enhance patient/resident comfort is unclear. Comfort is often considered to be merely the absence of discomfort. DESIGN: A critical ethnographic approach was used to facilitate understanding of nursing home residents' comfort. Fieldwork included 90 days of participant observation, interviews with 27 residents and 28 staff and extensive document examination. RESULTS: Comfort was multidimensional, idiosyncratic, dynamic and context dependent, rather than merely the absence of discomfort. This multidimensional nature meant residents could be 'betwixt and between' comfort and discomfort simultaneously. The constraints of one-size-fits all care delivery practices and the tensions inherent in communal living compounded residents' discomfort. CONCLUSIONS: Individualized care, based on comprehensive and accurate nursing assessment, is fundamental to the comfort of residents. RELEVANCE TO CLINICAL PRACTICE: Staff working in nursing homes must re examine their care delivery practices to ensure these do not disempower residents. Ensuring the comfort of each resident is a nursing priority and the key component of individualized care. PMID- 17462045 TI - Using action research to construct national evidence-based nursing care guidance for gerontological nursing. AB - AIM: This paper explores the development of a low-cost, involving methodology for constructing nursing-focused evidence-based national care guidance, known as Best Practice Statements, the intended users of which are gerontological nurses practising throughout Scotland. DESIGN: The Best Practice Statement construction methodology forms one cycle in a five-year longitudinal action research study that aims to achieve evidence-based nursing, facilitate professional networking to support practice development and promote the principles and practice of gerontological nursing. Achieving these aims involved designing a virtual Practice Development College. METHODS: A Community of Practice comprising practising gerontological nurses, expert advisors, academic teaching and research nurses collaborated in face-to-face meetings and in the virtual Practice Development College to delineate and refine the procedural model for Best Practice Statement construction. Focus groups, telephone interviews, analysis of on-line archives and documentary outputs formed the analytic dataset. RESULTS: Qualitative analysis indicated that, from the perspective of the community of practice, the emerging methodology facilitated the melding of knowledge sources reflecting the dominant evidence hierarchy with other forms of evidence valued by gerontological nurses, in the Best Practice Statement. RELEVANCE TO CLINICAL PRACTICE: Current methods of care guidance construction rarely address the concerns of nurses and the evidence from which guidelines are developed is narrowly defined with regard to inclusion and acceptability. In contrast this model focuses on nursing issues, embraces a wider definition of evidence and ensures that the published Best Practice Statements are credible and achievable in gerontological practice, where they are tested and refined as an inherent aspect of the development process. PMID- 17462046 TI - Exploring the facets of continence care: a continence survey of care homes for older people in Birmingham. AB - AIMS: This study aimed to explore continence prevalence, knowledge and care in Birmingham care homes, UK. BACKGROUND: There is an increasing need to respond to rising numbers of care home residents suffering from incontinence in the UK. A lack of staff knowledge, training and treatment options can make this problematic. METHOD: A survey was developed and sent to 186 care homes in Birmingham catering for those 65 years old and over. A freepost envelope was provided for return of the completed form. RESULTS: Sixty-six (35%) surveys were returned providing a representative sample. Eighty per cent of nursing home residents and 49% of residential home residents were reported incontinent. Half of the staff (53%) had some form of qualification in caring/nursing. Absorbent products were used to manage incontinence in over 50% of homes. Advanced age was cited as the main cause of incontinence. Sixty-eight per cent of homes used the continence advisory service and/or district nursing services. CONCLUSIONS: While results proved encouraging in relation to numbers of qualified staff there is a need for improved continence awareness. Focus on aetiology, assessment procedures and treatment options are needed. While external service use levels were encouraging there appeared to be a need for an exchange of knowledge between services to optimize care. RELEVANCE TO CLINICAL PRACTICE: While there is an existing body of knowledge relating to continence prevalence and management in nursing homes this research base is lacking for residential care settings. This survey addresses this imbalance by combining information from both settings. This paper also provides an insight into the components that have an impact upon continence promotion, prevention and management. By establishing a picture of current practice an indication of areas for improvement can be exposed. PMID- 17462047 TI - A survey of Chinese nurses' current knowledge of pain in older people. AB - AIM: To investigate the current level of Chinese nurses' knowledge of pain and pain management in older people. BACKGROUND: Most research about nurses' knowledge regarding pain has taken place in developed countries; however, limited research in this area has taken place in developing countries and particularly in China. METHODS: Registered Nurses (n = 621) in three different hospitals were surveyed with the questionnaire about pain and pain management with respect to older people. RESULTS: The finding showed that a significant knowledge deficit in this area exists. There were no significant differences among nurses in terms of education background, position and whether or not there was attendance in an educational session on pain management. However, there were statistically significant differences based on age; hospital of employment and clinical area in which employed. CONCLUSION: This survey suggested that nurses' knowledge of pain in older people should be improved. Basic and continuing education of nurses in this area should be enhanced and their active participation in pain management should be encouraged. RELEVANCE TO CLINICAL PRACTICE: The findings in this survey highlight a significant pain management knowledge deficit among the nurses in the clinical practice. The findings may help the nurses realize their knowledge deficit in this area and may also suggest curriculum changes for the nurses to improve pain management knowledge. PMID- 17462048 TI - Changing needs of Chinese family caregivers of stroke survivors. AB - AIM: To examine the changing needs of Chinese family caregivers before and after stroke survivors' discharge from hospital. BACKGROUND: Stroke ranks third as the most common cause of death in Hong Kong and is the leading cause of adult disability. Community care has been adopted as the major source of care for older people in Hong Kong and this has impacted on family caregivers. DESIGN AND METHODS: This is a descriptive-correlational design using a within-subjects design. The needs of 40 Chinese family caregivers who met the inclusion criteria were assessed before discharge and two weeks later using the Carer Assessment Scale, Cost of Care Index and one open-ended question. Modified Barthel Index measured the functional ability of stroke survivors. RESULTS: Family caregivers are able to anticipate most of their needs and to make provision to meet the basic practical needs before discharge. Although needs changed after discharge the four most important needs persisted. These were associated with emotional and psychological problems and financial difficulties. Discharge destinations made no difference to the total scores obtained using the above scales. CONCLUSIONS: This study provides information about need at a time of transition in an under researched population of Chinese caregivers. Assessment of need is important with Chinese family carers in order to identify focused interventions in a population, i.e. reluctant to make their needs known to professional services. More research about caregiving problems for Chinese family caregivers at the transition from hospital to community is required. RELEVANCE TO CLINICAL PRACTICE: Ongoing need assessment by nurses who are in regular contact with caregivers in hospital and community will enable appropriate interventions such as providing education and emotional support both before and after discharge to be offered to Chinese communities. PMID- 17462049 TI - Problem-solving experiences among elders living alone in eastern Taiwan. AB - AIM: To explore problem-solving experiences among older people living alone in eastern Taiwan. BACKGROUND: Elders living alone in Taiwan have a low participation rate in government social support programmes. Providing sufficient care for this population of elders requires knowing not only their problems, but also how and when they seek help solve their problems. Design. Qualitative research was used to gain a deeper understanding of the problem-solving experiences of older people living alone. METHODS: Elders living alone (n = 9) were recruited from a remote area of Taiwan. Data were gathered in audiotaped, semi-structured interviews and analysed by content analysis. RESULTS: Participants used both internal and external resources to solve problems through an appraisal process. Their internal resources included self-perception of health status, preventive coping strategies, flexible coping ability and being resigned to one's situation. Their external resources included both human and environmental resources. Based on their lived experience, participants appraised the usefulness of both internal and external resources before deciding whether to seek help from external resources. CONCLUSION: In Chinese society, maintaining a balance between internal and external resources is an important mechanism for elders in solving problems. RELEVANCE TO CLINICAL PRACTICE: Our findings may enhance policy makers' and healthcare providers' understanding of the problem solving experiences of elders who live alone in Taiwan. In practical terms, these findings can be used to improve the elderly people's use of support systems in their community. PMID- 17462050 TI - Weight increase in patients with dementia, and alteration in meal routines and meal environment after integrity promoting care. AB - AIMS: To follow weight changes in patients with moderate and severe dementia and analyse how these changes related to biological and psychological parameters after staff education and support in integrity promoting care. A further aim was to describe meal environment and routines relative to the intervention. BACKGROUND: Weight loss in patients with dementia and in particular Alzheimer's disease is common. The aetiology appears multifactorial with the meal environment and a decreased independence while eating among the factors. METHOD: Over a three month intervention period, an integrity-promoting care training programme was conducted with the staff of a long-term ward. Alzheimer's disease patients, 18 from an intervention ward and 15 from a control ward were included and possible effects were evaluated. Weighing was conducted at the start and after completion of the intervention. Weight changes were analysed in relation to psychological and biochemical parameters. In addition, the staff wrote diaries about, for example changes made in the environment and in their work. RESULTS: The most prominent difference observed was weight increases in 13 of 18 patients compared with two of 15 patients in the control ward. No weight changes were related to the type of dementia. The individual weight changes correlated significantly to changes in the intellectual functions. Relationships between weight change, increased motor function and increased appetite were non-significant. There was no significant relationship between weight changes and changes in biochemical parameters. According to the staff, increased contact with the patients and a more pleasant atmosphere resulted when the meal environment and routines were changed. RELEVANCE TO CLINICAL PRACTICE: Weight gain in patients with moderate and severe dementia was achieved by adjusting the meal environment to the individual's needs. Staff education was profitable, as increased competence seemed to promote individually adapted feeding situations. Ensuring good meal situations need to be given high priority. PMID- 17462052 TI - Treatment of undescended testes--time for a change in European traditions. PMID- 17462053 TI - Cryptorchidism: classification, prevalence and long-term consequences. AB - Undescended testis is a common finding in boys, and the majority of cases have no discernible aetiology. There are unexplained geographical differences and temporal trends in its prevalence. Cryptorchidism, especially bilateral, is associated with impaired spermatogenesis and endocrine function and increases the risk of testicular cancer. There is an urgent need to identify factors that adversely affect testicular development and optimize treatment. CONCLUSION: Cryptorchidism may reflect a primary testicular maldevelopment with long-term consequences. PMID- 17462054 TI - Studies of cryptorchidism in experimental animal models. AB - Undescended testes are the most common inborn anomaly in boys but our understanding of the causes is still sparse. Experimental models of cryptorchidism in various species of animals have been used by many investigators in order to gain a better understanding of the aetiology, pathophysiology and potential treatment of the disorder. This review presents what has been gained so far by studies using experimental animal models of cryptorchidism. CONCLUSIONS: Early orchidopexy in experimentally cryptorchid animals of different species prevents, at least partially, against testicular damage and infertility. This supports an early intervention strategy in the management of cryptorchidism in humans although animal data on the correct timing of such treatment cannot be directly translated into human clinical practice due to the species differences in pre and postnatal testis maturation. Treatment with pharmacological doses of hCG has poor efficacy in experimental animals and may cause adverse effects. Thus, clinically relevant doses of hCG causes acute inflammation-like changes in rodent testes and there are also indications that it may induce morphologically detectable permanent testicular damage. PMID- 17462055 TI - Development and descent of the testis in relation to cryptorchidism. AB - The testis descends in two phases. Animal studies suggest, that the transabdominal descent of the testis depends on the insulin-like hormone 3 (INSL3). Androgens are important in the inguinoscrotal testicular descent in animals and humans. In general, the cause of cryptorchidism is unknown and the aetiology is possibly multifactorial. Histological changes in cryptorchid testes demonstrate disturbed development. CONCLUSION: Since testicular descent is regulated by testis-derived hormones, cryptorchidism may reflect a functional defect of the testis. PMID- 17462056 TI - Efficacy and safety of hormonal treatment of cryptorchidism: current state of the art. AB - Meta-analyses of randomised trials using hCG or GnRH for treatment on testicular descent show in most studies overall efficacy of about 20%, less if retractile testes were excluded. In recent years a number of potentially serious side effects have been reported. CONCLUSION: Considering the efficacy and the possible side effects of the hormonal treatment, the general use of hCG and GnRH in the treatment of cryptorchidism cannot be further recommended. PMID- 17462057 TI - Resources allocation for neonatal critical care: the ethical debate in neonatology. PMID- 17462058 TI - Immediate maternal thermal response to skin-to-skin care of newborn. AB - AIM: To elucidate the immediate maternal thermal skin response when placing the neonate on the mothers' chest, skin-to-skin (STS). METHODS: Ugandan mothers with non-asphyxiated newborns after vaginal delivery (n = 39) were studied. Maternal skin and axillary temperatures were measured immediately before STS contact, then every 2 min for 20 min and finally 10 min after removing the neonate. Neonatal axillary/forehead temperatures were measured immediately before STS contact, and twice after initiating STS, followed by a measurement 10 min after the newborn had been removed. RESULTS: A rapid thermal response was demonstrated in maternal breast skin immediately after STS contact. It rose by 0.5 degrees C (p < 0.0001) on average the first 2 min after STS contact and fell by 0.5 degrees C 10 min after we had removed the neonate (p < 0.0001). Maternal axillary temperature also rose 2 min after initiation of STS (p < 0.0001) but stayed constant 10 min after removal of the newborn from the STS position. CONCLUSION: The findings indicate that there is a rapid maternal, thermal response to the positioning of the newborn STS. The tactile contact may elicit a maternal adaptation enhancing the warming of the newborn. Possible mechanisms include maternal autonomic nerve mediated skin vasodilatation. PMID- 17462059 TI - Neonatal end-of-life practice in a German perinatal centre. AB - AIM: To investigate the end-of-life practice in a large perinatal centre in Germany. METHODS: Retrospective chart review was performed in all neonates deceased in the delivery room (n = 31) and the neonatal intensive care unit (n = 47) between 2002 and 2004. RESULTS: Neonatal death was preceded by an end-of-life decision (EOLD) in 81% of cases in the delivery room and 83% in the neonatal intensive care unit. The majority of deceased neonates were born prematurely or with congenital malformation. Life-sustaining treatment was not initiated in 74% of the infants deceased in the delivery room. In the unit, 52% died after withdrawal of therapy. Mechanical ventilation was withdrawn most frequently (79% of cases). Futility and immediate death were common considerations in EOLD, but the infant's suffering and future quality of life also played a role. Parents were involved in EOLD-making in all but emergency cases. No active termination of life was performed. CONCLUSION: In our perinatal centre, the majority of neonatal deaths occurred after limitation of therapy. Treatment was actively withdrawn in half of the infants in the neonatal intensive care unit. Actual end-of-life practice in a large perinatal centre differs from the restrictive attitude towards EOLD reported for German neonatologists in previous surveys. PMID- 17462060 TI - Characteristics of severely malnourished under-five children hospitalized with diarrhoea, and their policy implications. AB - AIM: Identify clinical and nutritional features, and complications among severely malnourished, under-five children in an urban diarrhoeal disease facility in Bangladesh. METHODS: For this case-control design, children of both sexes, aged 0 59 months were studied. Severely (< -3 z-score) underweight, stunted or wasted constituted cases and those with better nutritional status (z-score > or = -3) constituted controls. RESULTS: During 2000-2005, of the total 6881 children, 1103 (16%) were severely underweight, 705 (11%) severely stunted and 217 (3%) severely wasted. In logistic regression analysis, severely underweight children were more likely to be older than 11 months (OR 3.7, 95% CI 3.1-4.3, p < 0.001), non breastfed (OR 1.5, 95% CI 1.3-1.8, p < 0.001), have illiterate mothers (OR 2.6, 95% CI 2.2-3.0, p < 0.001), non-sanitary toilet (OR 1.4, 95% CI 1.2-1.6, p < 0.001), a history of measles in preceding 6 months (OR 1.7, 95% CI 1.3-2.4, p = 0.001), dehydrating diarrhoea (OR 1.9, 95% CI 1.6-2.2, p < 0.001), abnormal findings in lung auscultation (OR 1.7, 95% CI 1.3-2.3, p < 0.001) and require hospitalization > or = 48 h (OR 2.2, 95% CI 1.8-2.5, p < 0.001). CONCLUSION: There thus is a need to incorporate appropriate, cost-effective and sustainable preventive strategies and improved management policies in the health systems as well as in social support systems in Bangladesh. PMID- 17462061 TI - The impact of recurrent abdominal pain: predictors of outcome in a large population cohort. AB - AIM: Recurrent abdominal pain is one of the commonest complaints of childhood, affecting approximately 10% of children at any one time. This study aims to describe the outcome of recurrent abdominal pain amongst 6-year-old children, and to identify risk factors (in both the children and their parents), which predict these outcomes. METHODS: 7128 6-year-old children in the Avon Longitudinal Study of Parents and Children (ALSPAC) were followed up for 1 year. The principle outcomes were abdominal pain, number of days missed from school in the preceding year and psychiatric diagnoses. RESULTS: Children with recurrent abdominal pain at age 6 years had higher rates of further abdominal pain [odds ratio 4.88 (95% CI: 4.01, 5.95)], school absence (10 or more days off school in a year) [OR 1.89 (1.53, 2.33)] and anxiety disorders [OR 2.36 (1.68, 3.32)] at age 7 years. Maternal anxiety was the most consistent predictor of subsequent adverse outcomes for these children. CONCLUSION: Children with recurrent abdominal pain are at significant and continuing risk of adverse functioning. It is important for clinicians to take a whole family perspective, and particularly to consider parental anxiety, when a child with recurrent abdominal pain presents to them. PMID- 17462062 TI - Satisfactory long-term results after Nissen fundoplication. AB - BACKGROUND: Complications and unsatisfactory long-term results after antireflux surgery in children have received increased attention. The aim of this study was to report parental assessment of outcome after Nissen fundoplication. METHODS: Ninety-three patients operated with primary Nissen fundoplication between 1990 and 2001 were included. Medical records were reviewed, and parents were interviewed for evaluation of postoperative results. RESULTS: Of the 93 children, 51 were neurologically impaired and 14 children had repaired esophageal atresia. Median follow-up time was 6.0 years. Total mortality for the patient population was 13%. Five children died as a complication of the surgery, whereas eight deaths were unrelated to the Nissen fundoplication. Ninety-two percent of the parents reported better well-being of the child after the Nissen fundoplication, and 83% were completely satisfied with the postoperative results. Pulmonary symptoms were reduced in 59%, and quality of sleep improved in 68% of the children. Nine children (10%) had been operated with a redo NF. CONCLUSION: The majority of parents were satisfied with the long term results of the Nissen fundoplication. PMID- 17462063 TI - The familial form of atrial septal defect. AB - BACKGROUND: The secundum atrial septal defect accounts for 10% of congenital heart disease. Familial occurrence is rare and may present as an isolated lesion or with conduction and skeletal abnormalities. Predisposing genes were described. OBJECTIVES: To evaluate familial defect's prevalence and associated anomalies and assess the yield of screening. METHODS: Family history, physical, electrocardiographic and echocardiographic evaluation of 286 ASD patients and families regarding heart disease, conduction and skeletal anomalies were performed. RESULTS: Eleven new families with familial defects were identified yielding 28 patients. The rate of transmission was 40-100%, suggestive of autosomal dominant inheritance. Parents were healthy in four families with multiple offspring with ASDs. Two families had ASDs with atrioventricular conduction abnormalities in five of six subjects, not requiring pacing. One subject had skeletal malformation. Ten patients had surgery, 12 had transcatheter ASD closure and six await treatment. CONCLUSIONS: In view of the high prevalence of familial occurrence of secundum ASD (10% of all ASD patients), we recommend screening all first degree relatives of ASD patients for cardiac, conduction and skeletal anomalies. Although a routine genetic investigation is not yet available, genetic patterns of inheritance may be compatible with autosomal dominant inheritance. Healthy parents of affected offspring may suggest a variable gene penetrance or past spontaneous ASD closure. Conduction anomalies may be present or may develop throughout life, and thus should be periodically screened for. PMID- 17462064 TI - Child behaviour after anaesthesia: associated risk factors. AB - AIM: To identify hospital care factors which are associated with problematic behaviours in children after hospitalization. METHOD: A cohort of 340 children ages 2-13 was studied in connection with elective procedures which included anaesthesia. DATA COLLECTED: sociodemographic, type of procedure, anaesthesia induction technique and premedication. Staff and parents assessed child anxiety at induction of anaesthesia, pain, anxiety and nausea in recovery room and hospital ward. Parents assessed their child's pain and nausea and the behaviour measured with the Post Hospital Behavioural Questionnaire two weeks after hospitalization. RESULTS: One-third (34.4 %) of the children developed at least one problematic behaviour, measured by the PHBQ subscales. Multiple logistic regression identified the following risk factors: age <5, pain at home but not at hospital, nausea, child anxiety at anaesthesia induction, postoperative nausea, postoperative distress, previous hospitalizations, living in a one adult family and having some previous problematic behaviours. Moderate-to-severe pain at home, but not at hospital, was associated with the greatest risk (OR 6.39 CI: 3.53 11.6). Previous anaesthesia, midazolam use in premedication and living in rural areas seemed to be protective factors. CONCLUSION: Pain at home but not in hospital is a strong risk factor for the onset or worsening of problematic behaviour after childhood hospitalization, which included anaesthesia. Proactive interventions are suggested to prevent this by improving pain treatment at home. PMID- 17462065 TI - A validation study of the Norwegian version of the Ages and Stages Questionnaires. AB - AIM: To explore the construct validity of the ASQ in relation to premature birth; maternal age, completeness of the family and level of education; and the child's gender. METHODS: Norwegian mothers' evaluations of 1172 children who completed age appropriate ASQs twice were statistically analysed by means of univariate and multivariate methods. RESULTS: The developmental level of girls is on average higher than that of boys at both assessments and there is a substantial increase between follow-up assessments in all ASQ scores except for gross motor skills. The mother's level of education; whether the child was born prematurely or at full-term; and the interaction between the child's gender and his/her mother's level of education are important factors relating to the child's developmental status in both assessments. CONCLUSION: The results that were derived by means of the Ages and Stages Questionnaires concerning the impact of maternal socio demographical factors, the child's gender, and premature birth upon early childhood development largely confirm the findings in the literature supporting the construct validity of the ASQ. The ASQ was found to be an effective diagnostic tool of developmental delay and/or disturbances. The development of gender dependent norms is recommended. PMID- 17462066 TI - Noncardiac comorbidities of congenital heart disease in children. AB - BACKGROUND: Noncardiac conditions may complicate heart disease care, contribute to the progression of the disease and alter the response to treatment. The aim of this prospective study was to evaluate the proportion of cardiac children with such significant conditions. METHODS: Of the 1058 children with congenital heart disease primarily diagnosed during a 10-year-period in one hospital, we identified those patients who had significant congenital and acquired comorbidities. RESULTS: Associated problems were diagnosed in 224 children (21.2%). Among them, 118 children (11.2%) had genetic or syndromic conditions, of which 38 had Trisomy 21. Six subspecialty areas accounted for the vast majority of the cases: neurology (n = 140), pulmonology (n = 36), orthopaedics (n = 26), nephro-urology (n = 19), gastroenterology (n = 14) and endocrinology (n = 13). The most frequent associated conditions were mental retardation, asthma, epilepsy and scoliosis. During the study period, death occurred in 46 of the children (4.3%) and was not related to the cardiac disease in 20 cases (1.9%). CONCLUSION: A substantial proportion of children with congenital heart disease have significant noncardiac comorbidities. Close collaboration between paediatric cardiologists and paediatricians of other subspecialties is imperative to optimize care for these children. PMID- 17462067 TI - Cognitive strengths and deficits in schoolchildren with ADHD. AB - BACKGROUND: Few studies provide detailed analyses of the various aspects of the entire cognitive profile of children with ADHD. MATERIAL AND METHODS: Cognitive test data were analysed for 10- to 11-year-old children with (1) ADHD, (2) subthreshold ADHD and (3) milder attention and/or learning problems, and compared with normative data. RESULTS: Thirty-two had ADHD and 10 met the criteria for subthreshold ADHD, prevalence rates of 5.4% and 1.6%, respectively. On a group level, children with ADHD/subthreshold ADHD, and those with milder attention and/or learning problems had almost identical cognitive profiles for the 13 subtests comprising the WISC III, with particularly low results on the arithmetic, coding, information and digit span subtests (ACID profile). When analyzed individually, a complete or incomplete ACID profile (three of four subtests) was equally common in children with ADHD/subthreshold ADHD and in children with milder problems, found in about 1/5. The relative strengths of both groups were in areas demanding logical thinking, reasoning and common sense. CONCLUSION: The specific ACID profile is as common in children with ADHD as in those with minor attention and/or learning problems. The cognitive weaknesses reflected in the ACID profile might play a role as an underlying factor in various developmental disorders. PMID- 17462068 TI - Case 2: A five-year-old girl with low-grade fever and pain in the leg. PMID- 17462071 TI - Long-term follow-up of a complicated crown fracture treated by partial pulpotomy. AB - AIM: To present the 7-year follow-up of a permanent incisor with complicated crown fracture, treated by partial pulpotomy. SUMMARY: A healthy permanent right central maxillary incisor with complicated crown fracture was treated by partial pulpotomy and reviewed clinically and radiographically for 7 years. At each recall, there was no spontaneous pain; the pulp showed signs of vitality and no periapical radiolucency developed. KEY LEARNING POINTS: Partial pulpotomy may be a successful permanent treatment in cases of complicated crown fracture. PMID- 17462077 TI - Viral vector vaccines make memory T cells against malaria. AB - Vaccines that comprise attenuated viral vectors encoding antigens from target pathogens generate potent T-cell responses. One such pathogen is malaria, and in particular the liver stage of its life cycle. Immunogenicity and efficacy studies in animals and humans have revealed the generation of memory T cells of both the central and effector phenotypes, depending on the viral vectors used in the malaria vaccination regime (viral species and serotype, combination and sequence for prime-boost) and suggest a divergence in their protective role. Being able to influence the memory T-cell make-up in a rational manner may allow us to develop more efficacious vaccines. PMID- 17462078 TI - Differential induction of CD94 and NKG2 in CD4 helper T cells. A consequence of influenza virus infection and interferon-gamma? AB - Influenza A virus causes worldwide epidemics and pandemics and the investigation of memory T helper (Th) cells that help maintain serological memory following infection is important for vaccine design. In this study we investigated CD94 and NKG2 gene expression in memory CD4 T-cell clones established from the spleens of C57BL/10 (H-2(b)) and BALB/c (H-2(d)) mice infected with influenza A virus (H3N2). CD94 and NKG2A/C/E proteins form heterodimeric membrane receptors that are involved in virus recognition. CD94 and NKG2 expression have been well characterized in natural killer (NK) and cytotoxic T cells. Despite CD94 being potentially an important marker for Th1 cells involved in virus infection, however, there has been little investigation of its expression or function in the CD4 T-cell lineage and no studies have looked at in-vivo-generated Th cells or memory cells. We show in this study that in-vivo-generated CD4 Th1 cells, but not Th2 cells, exhibited full-length CD94 and NKG2A gene expression following activation with viral peptide. For NKG2A, a novel 'short' (possibly redundant) truncated isoform was detectable in a Th2 cell clone. Another member of the NK receptor family, NKG2D, but not NKG2C or E, was also differentially expressed in Th1 cells. We show here that CD94 and NKG2A may exist as multiple isoforms with the potential to distinguish helper T-cell subsets. PMID- 17462079 TI - Decrease and dysfunction of dendritic cells correlate with impaired hepatitis C virus-specific CD4+ T-cell proliferation in patients with hepatitis C virus infection. AB - Through the production of stimulatory and suppressive cytokines, dendritic cells (DCs) regulate virus-specific immune responses that are crucial to virus eradication. To explore a possible role of DCs in the persistence of hepatitis C virus (HCV) infection, in this study we analysed peripheral blood DCs (PBDCs) in patients with chronic hepatitis C (CHC) compared with those in both healthy seronegative (HSN) controls and a group of subjects who had spontaneously resolved infection, defined as healthy HCV-seropositive (HSP), and we evaluated the relationships between PBDCs and HCV-specific CD4(+) T-cell reactivity. The number of PBDCs, their immunophenotype and expression of regulatory cytokines were evaluated by flow cytometry on whole-blood samples. HCV-specific CD4(+) T cell activation, proliferation and cytokine production were evaluated in cultures of peripheral blood mononuclear cells (PBMCs) stimulated in vitro with HCV peptides. We found that PBDCs from CHC subjects were numerically reduced and showed lower interleukin-12 (IL-12) and higher IL-10 expression than those from HSN controls. PBDCs from HSP subjects were similar to those from HSN controls. HCV-specific CD4(+) T-cell proliferation was less frequent and vigorous in CHC than in HSP patients and was directly related to the number of PBDCs and their IL 12 production but inversely related to their IL-10 production. Taken together, these results seem to suggest that cytokines of DC origin contribute to the regulation of HCV-specific immunity in CHC patients and indicate that PBDCs may represent a novel non-invasive tool for immune monitoring of these patients. PMID- 17462080 TI - Impact of the individual components of the metabolic syndrome and their different combinations on the prevalence of atherosclerotic vascular disease in type 2 diabetes: the Diabetes in Germany (DIG) study. AB - BACKGROUND: One of the major controversies surrounding the metabolic syndrome (MetS) in type 2 diabetes is whether its single components act synergistically as risk factors for atherosclerotic vascular disease (AVD). We aimed to answer this by evaluating the relationship, and its various combinations to AVD in comparison to single traits in a population-based study with type 2 diabetes in Germany. METHODS AND RESULTS: 4020 unselected patients with type 2 diabetes aged 35 - 80 years. MetS was: diabetes plus > or = 2 traits of the MetS by AHA/NHBLI definition.AVD was: history of myocardial infarction and/or coronary revascularization and/or stroke. The occurrence of AVD in relation to overall MetS/single traits/combinations was presented as OR (95% CI). Multiple logistic regression, including established cardiovascular risk factors, modeled their associations. The prevalence of overall MetS was 74.4% and the OR for AVD was 1.41 (1.12-1.78), which however was higher for hypertension as single trait (OR 4.76). Different combinations of MetS presented a wide range of ORs (0.47 to 10.90) and strong sex differences. Some clusters of MetS including hypertension and low HDL-cholesterol presented a higher risk factor than single traits or their sum, whereas the others out of 11 possible carried no increased AVD risk. Multiple logistic regression showed independent association between AVD and overall MetS. CONCLUSION: The overall MetS in type 2 diabetes comprises 11 heterogenous clusters of traits. Overall MetS increases the risk of AVD in type 2 diabetes and individual traits in some clusters with hypertension and low HDL cholesterol may act synergistically as risk factors particularly in women. PMID- 17462081 TI - When "no" might not quite mean "no"; the importance of informed and meaningful non-consent: results from a survey of individuals refusing participation in a health-related research project. AB - BACKGROUND: Low participation rates can lead to sampling bias, delays in completion and increased costs. Strategies to improve participation rates should address reasons for non-participation. However, most empirical research has focused on participants' motives rather than the reasons why non-participants refuse to take part. In this study we investigated the reasons why older people choose not to participate in a research project. METHODS: Follow-up study of people living in Tayside, Scotland who had opted-out of a cross-sectional survey on activities in retirement. Eight hundred and eighty seven people aged 65-84 years were invited to take part in a home-based cross-sectional survey. Of these, 471 refused to take part. Permission was obtained to follow-up 417 of the refusers. Demographic characteristics of people who refused to take part and the reasons they gave for not taking part were collected. RESULTS: 54% of those invited to take part in the original cross-sectional survey refused to do so. However, 61% of these individuals went on to participate in the follow-up study and provided reasons for their original refusal. For the vast majority of people initial non-participation did not reflect an objection to participating in research in principle but frequently stemmed from barriers or misunderstandings about the nature or process of the project itself. Only 28% indicated that they were "not interested in research". The meaningfulness of expressions of non consent may therefore be called into question. Hierarchical log-linear modelling showed that refusal was independently influenced by age, gender and social class. However, this response pattern was different for the follow-up study in which reasons for non-participation in the first survey were sought. This difference in pattern and response rates supports the likely importance of recruitment issues that are research and context specific. CONCLUSION: An expression of non-consent does not necessarily mean that a fully informed evaluation of the pros and cons of participation and non-participation has taken place. The meaningfulness of expressions of non-consent may therefore be a cause for concern and should be subject to further research. Many reasons for non-participation may be specific to a particular research topic or population. Information sheets should reflect this by going beyond standardised guidelines for their design and instead proactively seek out and address areas of concern or potential misunderstanding. The use of established behavioural theory in their design could also be considered. PMID- 17462083 TI - Effects of risk-based multifactorial fall prevention on health-related quality of life among the community-dwelling aged: a randomized controlled trial. AB - BACKGROUND: This study aimed to assess the effects of a risk-based, multifactorial fall prevention programme on health-related quality of life among the community-dwelling aged who had fallen at least once during the previous 12 months. METHODS: The study is a part of a single-centre, risk-based, multifactorial randomised controlled trial. The intervention lasted for 12 months and consisted of a geriatric assessment, guidance and treatment, individual instruction in fall prevention, group exercise, lectures on themes related to falling, psychosocial group activities and home exercise. Of the total study population (n = 591, 97% of eligible subjects), 513(251 in the intervention group and 262 in the control group) participated in this study. The effect of the intervention on quality of life was measured using the 15D health-related quality of life instrument consisting of 15 dimensions. The data were analysed using the chi-square test or Fisher's exact test, the Mann-Whitney U-test and logistic regression. RESULTS: In men, the results showed significant differences in the changes between the intervention and control groups in depression (p = 0.017) and distress (p = 0.029) and marginally significant differences in usual activities (p = 0.058) and sexual activity (p = 0.051). In women, significant differences in the changes between the groups were found in usual activities (p = 0.005) and discomfort/symptoms (p = 0.047). For the subjects aged 65 to 74 years, significant differences in the changes between the groups were seen in distress (p = 0.037) among men and in usual activities (p = 0.011) among women. All improvements were in favour of the intervention group. CONCLUSION: Fall prevention produced positive effects on some dimensions of health-related quality of life in the community-dwelling aged. Men benefited more than women. PMID- 17462082 TI - Food-associated cues alter forebrain functional connectivity as assessed with immediate early gene and proenkephalin expression. AB - BACKGROUND: Cues predictive of food availability are powerful modulators of appetite as well as food-seeking and ingestive behaviors. The neurobiological underpinnings of these conditioned responses are not well understood. Monitoring regional immediate early gene expression is a method used to assess alterations in neuronal metabolism resulting from upstream intracellular and extracellular signaling. Furthermore, assessing the expression of multiple immediate early genes offers a window onto the possible sequelae of exposure to food cues, since the function of each gene differs. We used immediate early gene and proenkephalin expression as a means of assessing food cue-elicited regional activation and alterations in functional connectivity within the forebrain. RESULTS: Contextual cues associated with palatable food elicited conditioned motor activation and corticosterone release in rats. This motivational state was associated with increased transcription of the activity-regulated genes homer1a, arc, zif268, ngfi-b and c-fos in corticolimbic, thalamic and hypothalamic areas and of proenkephalin within striatal regions. Furthermore, the functional connectivity elicited by food cues, as assessed by an inter-regional multigene-expression correlation method, differed substantially from that elicited by neutral cues. Specifically, food cues increased cortical engagement of the striatum, and within the nucleus accumbens, shifted correlations away from the shell towards the core. Exposure to the food-associated context also induced correlated gene expression between corticostriatal networks and the basolateral amygdala, an area critical for learning and responding to the incentive value of sensory stimuli. This increased corticostriatal-amygdalar functional connectivity was absent in the control group exposed to innocuous cues. CONCLUSION: The results implicate correlated activity between the cortex and the striatum, especially the nucleus accumbens core and the basolateral amygdala, in the generation of a conditioned motivated state that may promote excessive food intake. The upregulation of a number of genes in unique patterns within corticostriatal, thalamic, and hypothalamic networks suggests that food cues are capable of powerfully altering neuronal processing in areas mediating the integration of emotion, cognition, arousal, and the regulation of energy balance. As many of these genes play a role in plasticity, their upregulation within these circuits may also indicate the neuroanatomic and transcriptional correlates of extinction learning. PMID- 17462084 TI - The Consumer Quality Index Hip Knee Questionnaire measuring patients' experiences with quality of care after a total hip or knee arthroplasty. AB - BACKGROUND: The Dutch Consumer Quality Index Hip Knee Questionnaire (CQI Hip Knee) was used to assess patients' experiences with and evaluations of quality of care after a total hip (THA) or total knee arthroplasty (TKA). The aim of this study is to evaluate the construct validity and internal consistency reliability of this new instrument and to assess its ability to measure differences in quality of care between hospitals. METHODS: Survey data of 1,675 subjects who underwent a THA or TKA were used to evaluate the psychometric properties. Exploratory factor analyses were performed and item-total correlations and inter factor correlations were calculated to assess the construct validity of the instrument. Reliability analyses included tests of internal consistency (Cronbach's alpha coefficients). Finally, multilevel analyses were performed to assess the ability of the instrument to discriminate between hospitals in quality of care. RESULTS: Exploratory factor analyses indicated that the survey consisted of 21 items measuring five aspects of care (i.e. communication with nurses, communication with doctors, communication with general practitioner, communication about new medication, and pain control). Cronbach's alpha coefficients ranged from 0.76 to 0.90 indicating good internal consistency. The survey's ability to discriminate between hospitals was partly supported by multilevel analysis. Two scales (i.e. communication with nurses and communication with doctors) were able to measure differences between hospitals with respect to patients' experiences with quality of care. Logistic multilevel analyses indicated that hospitals explained part of the variation between patients in receiving information. CONCLUSION: These findings suggest that the CQI Hip Knee is reliable and valid for use in Dutch health care. Health care providers or health plans can use this survey to measure patients' experiences with hospital care and to identify variations in care between hospitals. PMID- 17462085 TI - Vitamin D inadequacy in Belgian postmenopausal osteoporotic women. AB - BACKGROUND: Inadequate serum vitamin D [25(OH)D] concentrations are associated with secondary hyperparathyroidism, increased bone turnover and bone loss, which increase fracture risk. The objective of this study is to assess the prevalence of inadequate serum 25(OH)D concentrations in postmenopausal Belgian women. Opinions with regard to the definition of vitamin D deficiency and adequate vitamin D status vary widely and there are no clear international agreements on what constitute adequate concentrations of vitamin D. METHODS: Assessment of 25 hydroxyvitamin D [25(OH)D] and parathyroid hormone was performed in 1195 Belgian postmenopausal women aged over 50 years. Main analysis has been performed in the whole study population and according to the previous use of vitamin D and calcium supplements. Four cut-offs of 25(OH)D inadequacy were fixed : < 80 nmol/L, <75 nmol/L, < 50 nmol/L and < 30 nmol/L. RESULTS: Mean (SD) age of the patients was 76.9 (7.5) years, body mass index was 25.7 (4.5) kg/m2. Concentrations of 25(OH)D were 52.5 (21.4) nmol/L. In the whole study population, the prevalence of 25(OH)D inadequacy was 91.3 %, 87.5 %, 43.1 % and 15.9% when considering cut-offs of 80, 75, 50 and 30 nmol/L, respectively. Women who used vitamin D supplements, alone or combined with calcium supplements, had higher concentrations of 25(OH)D than non-users. Significant inverse correlations were found between age/serum PTH and serum 25(OH)D (r = -0.23/r = -0.31) and also between age/serum PTH and femoral neck BMD (r = -0.29/r = -0.15). There is a significant positive relation between age and PTH (r = 0.16), serum 25(OH)D and femoral neck BMD (r = 0.07). (P < 0.05)Vitamin D concentrations varied with the season of sampling but did not reach statistical significance (P = 0.09). CONCLUSION: This study points out a high prevalence of vitamin D inadequacy in Belgian postmenopausal osteoporotic women, even among subjects receiving vitamin D supplements. PMID- 17462086 TI - Toward the automated generation of genome-scale metabolic networks in the SEED. AB - BACKGROUND: Current methods for the automated generation of genome-scale metabolic networks focus on genome annotation and preliminary biochemical reaction network assembly, but do not adequately address the process of identifying and filling gaps in the reaction network, and verifying that the network is suitable for systems level analysis. Thus, current methods are only sufficient for generating draft-quality networks, and refinement of the reaction network is still largely a manual, labor-intensive process. RESULTS: We have developed a method for generating genome-scale metabolic networks that produces substantially complete reaction networks, suitable for systems level analysis. Our method partitions the reaction space of central and intermediary metabolism into discrete, interconnected components that can be assembled and verified in isolation from each other, and then integrated and verified at the level of their interconnectivity. We have developed a database of components that are common across organisms, and have created tools for automatically assembling appropriate components for a particular organism based on the metabolic pathways encoded in the organism's genome. This focuses manual efforts on that portion of an organism's metabolism that is not yet represented in the database. We have demonstrated the efficacy of our method by reverse-engineering and automatically regenerating the reaction network from a published genome-scale metabolic model for Staphylococcus aureus. Additionally, we have verified that our method capitalizes on the database of common reaction network components created for S. aureus, by using these components to generate substantially complete reconstructions of the reaction networks from three other published metabolic models (Escherichia coli, Helicobacter pylori, and Lactococcus lactis). We have implemented our tools and database within the SEED, an open-source software environment for comparative genome annotation and analysis. CONCLUSION: Our method sets the stage for the automated generation of substantially complete metabolic networks for over 400 complete genome sequences currently in the SEED. With each genome that is processed using our tools, the database of common components grows to cover more of the diversity of metabolic pathways. This increases the likelihood that components of reaction networks for subsequently processed genomes can be retrieved from the database, rather than assembled and verified manually. PMID- 17462087 TI - Treatment-seeking behaviour for febrile illness in an area of seasonal malaria transmission in rural Ethiopia. AB - BACKGROUND: Very little is known about the management of malaria and treatment seeking patterns among children and adults in areas of seasonal malaria transmission particularly in east Africa. OBJECTIVES: The aim of this study was to assess treatment-seeking behaviour for reported malaria among all age groups in an area of seasonal transmission. METHODS: A community-based cross-sectional study was carried out among 2,253 households in 12 randomly selected rural kebeles in Adami Tulu district in south-central Ethiopia, during October-November 2003, using a pre-tested interviewer-administered structured questionnaire. RESULTS: Reported malaria was 14% among 12,225 people assessed during the last 14 days. Family/self-diagnosis was most common and the main first responses included visiting village-based community health workers (CHWs) (33%), public health facility (23%) and private clinic (17%). Home treatment was the least reported first response (3%). Only 13% had sought treatment within the first 24 hours of symptom onset. Early treatment-seeking pattern was reported among those who visited CHWs and practiced home treatment, with more delays among public facility users. Treatment-seeking behaviour was similar in all age groups. CONCLUSION: A considerable proportion of visits were made to CHWs and private providers, necessitating the importance of strengthening both community-based interventions and peripheral public and private facilities. Finally, the community should be informed and educated about the importance of early diagnosis and prompt treatment with effective antimalarials. PMID- 17462088 TI - Platelet adhesion onto immobilized fibrinogen under arterial and venous in-vitro flow conditions does not significantly differ between men and women. AB - BACKGROUND: Gender-related differences in incidence of arterial thrombosis have been a focus of interest for years. The platelet integrin alphaIIbbeta3 is primarily responsible for the interaction between platelets and fibrinogen and consecutive thrombus growth. In this study, we evaluated platelet adhesion onto immobilized fibrinogen under venous and arterial flow conditions in men and women. METHODS: Platelets in whole anticoagulated blood were labelled with the fluorescence dye Mepacrine and perfused through the rectangular flow chamber over glass cover slips coated with fibrinogen (shear rates of 50 s-1, 500 s-1 and 1500 s-1). A fluorescence laser-scan microscope was used for visualisation and quantification of platelet adhesion at 15 seconds, 1 and 5 minutes after the start of perfusion. RESULTS: During perfusion, the platelet adhesion linearly increased in regard to exposition time and shear rate. After five minutes of perfusion the platelet adhesion onto immobilized fibrinogen showed no significant gender related difference, neither at 50 s-1 nor at 500 s-1 and 1500 s-1 (p > 0.05), respectively. No significant difference in platelet adhesion onto immobilized fibrinogen, in regard to the menopausal status, was either observed (p > 0.05). CONCLUSION: In our in vitro experimental system, hormonal differences between men and women did not influence platelet adhesion onto immobilized fibrinogen, neither under venous nor under arterial rheological conditions. PMID- 17462089 TI - Cultural diversity teaching and issues of uncertainty: the findings of a qualitative study. AB - BACKGROUND: There is considerable ambiguity in the subjective dimensions that comprise much of the relational dynamic of the clinical encounter. Comfort with this ambiguity, and recognition of the potential uncertainty of particular domains of medicine (e.g.--cultural factors of illness expression, value bias in diagnoses, etc) is an important facet of medical education. This paper begins by defining ambiguity and uncertainty as relevant to clinical practice. Studies have shown differing patterns of students' tolerance for ambiguity and uncertainty that appear to reflect extant attitudinal predispositions toward technology, objectivity, culture, value- and theory-ladeness, and the need for self examination. This paper reports on those findings specifically related to the theme of uncertainty as relevant to teaching about cultural diversity. Its focus is to identify how and where the theme of certainty arose in the teaching and learning of cultural diversity, what were the attitudes toward this theme and topic, and how these attitudes and responses reflect and inform this area of medical pedagogy. METHODS: A semi-structured interview was undertaken with 61 stakeholders (including policymakers, diversity teachers, students and users). The data were analysed and themes identified. RESULTS: There were diverse views about what the term cultural diversity means and what should constitute the cultural diversity curriculum. There was a need to provide certainty in teaching cultural diversity with diversity teachers feeling under considerable pressure to provide information. Students discomfort with uncertainty was felt to drive cultural diversity teaching towards factual emphasis rather than reflection or taking a patient centred approach. CONCLUSION: Students and faculty may feel that cultural diversity teaching is more about how to avoid professional, medico-legal pitfalls, rather than improving the patient experience or the patient-physician relationship. There may be pressure to imbue cultural diversity issues with levels of objectivity and certainty representative of other aspects of the medical curriculum (e.g.--biochemistry). This may reflect a particular selection bias for students with a technocentric orientation. Inadvertently, medical education may enhance this bias through training effects, and accommodate disregard for subjectivity, over-reliance upon technology and thereby foster incorrect assumptions of objective certainty. We opine that it is important to teach students that technology cannot guarantee certainty, and that dealing with subjectivity, diversity, ambiguity and uncertainty is inseparable from the personal dimension of medicine as moral enterprise. Uncertainty is inherent in cultural diversity so this part of the curriculum provides an opportunity to address the issue as it relates to patient care. PMID- 17462090 TI - Failure of recombinant factor VIIa in a patient with severe polymicrobial sepsis and postoperative uncontrolled intraabdominal bleeding. AB - BACKGROUND: This report discusses a case of unsuccessful treatment with recombinant factor VIIa (rFVIIa) in off-label use. The need for international guidelines concerning the off-label use of rFVIIa is outlined as well as the need for methods to control the efficacy of rFVIIa objectively. CASE PRESENTATION: 54 year old male with severe polymicrobial sepsis due to a perforated diverticulitis of the sigmoid colon and consecutive overt disseminated intravascular coagulation. He suffered severe intraabdominal bleeding after abdominal surgery despite conventional haemostatic support. Repeated applications of factor VIIa temporarily improved coagulation essays but did not stop clinical bleeding. The patient died in multiorgan failure due to septic and haemorrhagic shock. CONCLUSION: Off-label use of rFVIIa could result in more side effects than could be expected from literature because of a publication bias. However for most off label applications large prospective, randomised and controlled trials to confirm the positive findings are missing. For the future, not only guidelines concerning the off-label use of rFVIIa are urgently needed but also guidelines for monitoring the efficacy of rFVIIa. PMID- 17462091 TI - Management of headache disorders: design of a randomised clinical trial screening for prognostic patient characteristics. AB - BACKGROUND: Treatment of headache disorders is not always optimal. Patients are treated in multiple ways, and the lack of scientific arguments for referral and the insufficient implementation of guidelines result in unclear treatment strategies. The coexistence of headache and neck pain can lead to the referral to a musculoskeletal physiotherapist. This treatment can only be successful if an underlying cervical segmental dysfunction is present. In such cases a physical treatment can be a valuable option that should be considered. The aim of this study is to identify prognostic therapeutic patient characteristics and to increase the number of correct physiotherapy referrals. METHODS/DESIGN: This trial is designed to identify patient characteristics which can influence the prognosis of the patient. Patients with recurrent headache and co-existent neck pain are recruited via a multicenter setup. After screening for eligibility, subjects are tested at baseline and randomly allocated to one of two treatment groups. Testing includes the administering of questionnaires (a Headache Diagnosis Questionnaire, Headache Inventory List and the Headache Impact Test (HIT-6)) and physical tests (Thermal Stimuli, Manual Cervical Spine Examination and Pressure Algometry). Treatment groups are a usual care group (UC) administered by the General Practitioner (GP) and a usual care plus musculoskeletal physiotherapy treatment group (UCMT). UC is based on the Dutch GP Guideline for Headache. UCMT consists of the UC plus a combination of exercises and spinal cervical mobilisations. Follow-up measurements consist of the completion of the Headache Inventory List, the HIT-6 and scoring of the global perceived effect (GPE). The latter allowing the distinction between responders (positive effect) and non-responders (no effect or worse). Logistic regression analysis will be used to identify the specific patient characteristics of the responders and the non-responders. The additional value of the musculoskeletal physiotherapy will be examined. Follow-up measurements up to 52 weeks are scheduled. DISCUSSION: This trial aims to identify prognostic patient characteristics, in order to supply a useful diagnostic tool for all health care workers, dealing with headache sufferers. PMID- 17462092 TI - Assessment of the toll-like receptor 4 Asp299Gly, Thr399Ile and interleukin-8 251 polymorphisms in the risk for the development of distal gastric cancer. AB - BACKGROUND: The intensity of the inflammation induced by Helicobacter pylori colonization is associated with the development of distal gastric cancer (GC). The host response to H. pylori has been related to genetic polymorphisms that influence both innate and adaptive immune responses.Our aim was to investigate whether the presence of the TLR4 Asp299Gly, TLR4 Thr399Ile and IL-8-251 A/T polymorphisms had any influence in the development of distal GC in a Mexican population. METHODS: We studied 337 patients that were divided in two groups: 78 patients with histologically confirmed distal GC and 259 non-cancer controls. The presence of H. pylori in the control population was defined by positive results of at least two of four diagnostic tests: serology, histology, rapid urease test and culture. Human DNA was purified and genotyped for TLR4 Asp299Gly polymorphism by pyrosequencing, for TLR4 Thr399Ile by PCR-RFLP and for IL8-251 by the amplification refractory mutation system (ARMS)-PCR. RESULTS: The non-cancer control group was found to be in Hardy-Weinberg equilibrium at the polymorphic loci studied (chi-square H-W = 0.58 for IL8-251, 0.42 for TLR4 Asp299Gly and 0.17 for TLR4 Thr399Ile). The frequencies of mutated alleles (homozygous plus heterozygous) were compared between cases and controls. We found no significant difference for TLR4- Asp299Gly [the 7.7% of distal GC patients and 7.7 % non cancer controls (p = 0.82)] and for TLR4 Thr399Ile [the 1.3% of GC patients and the 5% of the control population (p = 0.2)]. In contrast, for IL-8-251 A/T, 80.77% of the GC patients and 66.4% in the control group age and gender matched had at least one copy of mutated allele (OR = 2.12, 95% CI = 1.1-4.2) (p = 0.023). CONCLUSION: This study showed that the IL8-251*A allele could be related to the development of distal gastric cancer in this Mexican population. PMID- 17462093 TI - Impact of welfare cheque issue days on a service for those intoxicated in public. AB - In British Columbia (BC), the Ministry of Human Resources issues welfare cheques to eligible recipients monthly on the last Wednesday of each month. Previous studies have indicated that there are significant increases in hospital admission, ED admission, 911 calls and deaths shortly after the distribution of the monthly welfare cheques. The objective of this analysis was to rigorously examine the impact of welfare cheque issue dates on admission to the Sobering Unit (SU), a service for the publicly intoxicated, in Vancouver, Canada. Data on 1234 consecutive admissions to the SU over a 7-month period were assessed, and the average number of daily admissions on each of the 7 days of the welfare cheque issue week and similar weekdays in other weeks were compared. A Wilcoxon rank-sum test was performed for the comparisons. Our results showed that there were significant increases in the number of admissions on the 3 days starting with "Welfare Wednesday" compared to the similar weekdays in other weeks (Welfare Wednesday vs. other Wednesdays: 8.7 vs. 5.1, p = 0.02; Welfare Thursdays vs. other Thursdays: 9.6 vs. 5.3, p = 0.02; Welfare Fridays vs. other Fridays: 8.6 vs. 5.7, p = 0.04). The demonstrated impact of welfare cheque issue dates is an important consideration for the re-design, staffing and resource allocation of services for withdrawal management and potentially for other services offered to this population. PMID- 17462094 TI - Cross-sectional validation of the Aging Perceptions Questionnaire: a multidimensional instrument for assessing self-perceptions of aging. AB - BACKGROUND: Self-perceptions of aging have been implicated as independent predictors of functional disability and mortality in older adults. In spite of this, research on self-perceptions of aging is limited. One reason for this is the absence of adequate measures. Specifically, there is a need to develop a measure that is theoretically-derived, has good psychometric properties, and is multidimensional in nature. The present research seeks to address this need by adopting the Self-Regulation Model as a framework and using it to develop a comprehensive, multi-dimensional instrument for assessing self-perceptions of aging. This study describes the validation of this newly-developed instrument, the Aging Perceptions Questionnaire (APQ). METHODS: Participants were 2,033 randomly selected community-dwelling older (+65 yrs) Irish adults who completed the APQ alongside measures of physical and psychological health. The APQ assesses self-perceptions of aging along eight distinct domains or subscales; seven of these examine views about own aging, these are: timeline chronic, timeline cyclical, consequences positive, consequences negative, control positive, control negative, and emotional representations; the eighth domain is the identity domain and this examines the experience of health-related changes. RESULTS: Mokken scale analysis showed that the majority of items within the views about aging subscales were strongly scalable. Confirmatory factor analysis also indicated that the model provided a good fit for the data. Overall, subscales had good internal reliabilities. Hierarchical linear regression was conducted to investigate the independent contribution of APQ subscales to physical and psychological health and in doing so determine the construct validity of the APQ. Results showed that self-perceptions of aging were independently related to physical and psychological health. Mediation testing also supported a role for self perceptions of aging as partial mediators in the relationship between indices of physical functioning and physical and psychological health outcomes. CONCLUSION: Findings support the complex and multifaceted nature of the aging experience. The good internal reliability and construct validity of the subscales suggests that the APQ is a promising instrument that can enable a theoretically informed, multidimensional assessment of self-perceptions of aging. The potential role of self-perceptions of aging in facilitating physical and psychological health in later life is also highlighted. PMID- 17462095 TI - Noninvasive and invasive evaluation of cardiac dysfunction in experimental diabetes in rodents. AB - BACKGROUND: Because cardiomyopathy is the leading cause of death in diabetic patients, the determination of myocardial function in diabetes mellitus is essential. In the present study, we provide an integrated approach, using noninvasive echocardiography and invasive hemodynamics to assess early changes in myocardial function of diabetic rats. METHODS: Diabetes was induced by streptozotocin injection (STZ, 50 mg/kg). After 30 days, echocardiography (noninvasive) at rest and invasive left ventricular (LV) cannulation at rest, during and after volume overload, were performed in diabetic (D, N = 7) and control rats (C, N = 7). The Student t test was performed to compare metabolic and echocardiographic differences between groups at 30 days. ANOVA was used to compare LV invasive measurements, followed by the Student-Newman-Keuls test. Differences were considered significant at P < 0.05 for all tests. RESULTS: Diabetes impaired LV systolic function expressed by reduced fractional shortening, ejection fraction, and velocity of circumferential fiber shortening compared with that in the control group. The diabetic LV diastolic dysfunction was evidenced by diminished E-waves and increased A-waves and isovolumic relaxation time. The myocardial performance index was greater in diabetic compared with control rats, indicating impairment in diastolic and systolic function. The LV systolic pressure was reduced and the LV end-diastolic pressure was increased at rest in diabetic rats. The volume overload increased LVEDP in both groups, while LVEDP remained increased after volume overload only in diabetic rats. CONCLUSION: These results suggest that STZ-diabetes induces systolic and diastolic dysfunction at rest, and reduces the capacity for cardiac adjustment to volume overload. In addition, it was also demonstrated that rodent echocardiography can be a useful, clinically relevant tool for the study of initial diabetic cardiomyopathy manifestations in asymptomatic patients. PMID- 17462096 TI - Depression and the nature of Trinidadian family practice: a cross-sectional study. AB - BACKGROUND: Depression is the most common mental disorder; in an ambulatory-care setting 5 to 10% of patients meet the criteria for major depression. Despite extensive documentation in primary care internationally, Trinidadian studies published on depression have been primarily hospital-based and focussed on suicide. The objectives of this study were to determine the prevalence of depression, the variables associated with depression and the commonest reason for the encounter (RFE) among adult patients attending Trinidadian fee-for-service family practice? METHODS: This was a cross-sectional descriptive survey of consecutive patients taken from a stratified random sample of family practices in the north-west region of Trinidad. To measure depression the Zung scale was modified for use as a brief diagnostic tool. This modified Zung scale, when tested against a psychiatric interview, revealed that at a cut off point of 60, the scale had a specificity of 94% (95% CI 87-100), a sensitivity of 60% (95% CI 45-75), and a likelihood ratio for a positive test result of 10 (95% CI 6-42). RESULTS: 508 patients from 28 practices participated; a response rate of 85%. Participants were primarily younger 18-49 years (66.7%), female (69.5%), and educated, with 72.8% having received a secondary school, technical school or university education. Sixty-five (12.8%) of the respondents (95% CI 9.9-15.7) were determined to be depressed. Chi-square analysis revealed no statistically significant association between depression and age, ethnicity, education levels, occupation or marital status (p > 0.05). Binary logistic regression indicated that the likelihood of being depressed (p < 0.05) decreased with the increasing age of the patient and was inversely proportional to patient's achieved level of education; and that patients not presently in a relationship were more likely to be depressed than patients who were currently in a relationship. The 508 participants had 630 RFE, with 'check-ups' (17.5%) being the commonest, followed by joint pains (13.4%) and upper respiratory infections (10.5%). CONCLUSION: The Trinidadian family physician has to maintain a high index of suspicion in the knowledge that as many as one of every eight adult patients may be depressed and that younger patients of lower educational status who were not currently in a relationship were more likely to be depressed. PMID- 17462097 TI - Thiazolidinedione induced thyroid associated orbitopathy. AB - BACKGROUND: To describe an episode of thyroid associated orbitopathy (TAO) following the initiation of thiazolidinedione (TZD). CASE PRESENTATION: We report a female patient with a history of Graves' disease and stabilised thyroid associated orbitopathy for 2.5 years, who experienced rapid progression of TAO after the initiation of thiazolidinedione for glycemic control. Following the discontinuation of TZD, the patient experienced subsequent stabilisation of disease and normalization of vision. The medical history, ophthalmic findings, and clinical course are discussed. CONCLUSION: Thiazolidinediones may exacerbate TAO, and this should be taken into consideration when selecting treatment for diabetic patients with a history of autoimmune thyroid disorders. PMID- 17462098 TI - Global gene expression analysis and regulation of the principal genes expressed in bovine placenta in relation to the transcription factor AP-2 family. AB - BACKGROUND: Cell-cell communication is an important factor in feto-maternal units during placentogenesis. The placenta produces pivotal hormones and cytokines for communication between cotyledonary villi and the maternal caruncle. Gene expression in bovine placenta throughout pregnancy was comprehensively screened by a cDNA microarray, and we searched for a common transcription factor in a gene cluster that showed increasing expression throughout gestation in cotyledonary villi and caruncle. METHODS: Placentomal tissues (villi and caruncle) were collected from Day 25 to Day 250 of gestation for microarray analysis. Global gene expression profiles were analyzed using the k-means clustering method. A consensus sequence cis-element that may control up-regulated genes in a characteristic cluster was examined in silico. The quantitative expression and localization of a specific transcription factor were investigated in each tissue using quantitative real-time RT-PCR and in situ hybridization. RESULTS: The microarray expression profiles were classified into ten clusters. The genes with most markedly increased expression became concentrated in cluster 2 as gestation proceeded. Cluster 2 included placental lactogen (CSH1), pregnancy-associated glycoprotein-1 (PAG1), and sulfotransferase family 1E estrogen-preferring member 1 (SULT1E1), which were mainly detected in giant trophoblast binucleate cells (BNC). Consensus sequence analysis identified transcription factor AP-2 binding sites in some genes in this cluster. Quantitative real-time RT-PCR analysis confirmed that high level expression of transcription factor AP-2 alpha (TFAP2A) was common to cluster 2 genes during gestation. In contrast, the expression level of another AP-2 family gene, transcription factor AP-2 beta (TFAP2B), was extremely low over the same period. Another gene of the family, transcription factor AP-2 gamma (TFAP2C), was expressed at medium level compared with TFAP2A and TFAP2B. In situ hybridization showed that TFAP2A, TFAP2B and TFAP2C mRNAs were localized in trophoblast cells but were expressed by different cells. TFAP2A was expressed in cotyledonary epithelial cells including BNC, TFAP2B was specifically expressed in BNC, and TFAP2C in mononucleate cells. CONCLUSION: We detected gestational-stage-specific gene expression profiles in bovine placentomes using a combination of microarray and in silico analysis. In silico analysis indicated that the AP-2 family may be a consensus regulator for the gene cluster that characteristically appears in bovine placenta as gestation progresses. In particular, TFAP2A and TFAP2B may be involved in regulating binucleate cell-specific genes such as CSH1, some PAG or SULT1E1. These results suggest that the AP-2 family is a specific transcription factor for clusters of crucial placental genes. This is the first evidence that TFAP2A may regulate the differentiation and specific functions of BNC in bovine placenta. PMID- 17462099 TI - Letter to the editor: healthy alternatives to trans fats. AB - Consumption of trans fats is associated with an increase of cardiovascular disease (CVD) risk. To comply with regulatory policies and public health authorities recommendations, trans fats should be replaced in food products. The study by Sundram et al. (Nutrition & Metabolism 2007, 4:3) reporting the effect on CVD risk factors of interesterified fat (IE) and partially hydrogenated soybean oil (PHSO) compared to palm olein (POL) has been critically analyzed. The study design and in particular the composition of the tested fats was not suitable to properly answer the question raised regarding the effect of alternative ingredients to trans fats on plasma lipids. The observed effects are divergent with predicted data derived from the literature model consolidated using the individual results of 60 randomized clinical trials. The results of the study published by Sundram and co-workers have to be considered with awareness. PMID- 17462100 TI - Which health-related quality of life score? A comparison of alternative utility measures in patients with Type 2 diabetes in the ADVANCE trial. AB - BACKGROUND: Diabetes has a high burden of illness both in life years lost and in disability through related co-morbidities. Accurate assessment of the non mortality burden requires appropriate health-related quality of life and summary utility measures of which there are several contenders. The study aimed to measure the impact of diabetes on various health-related quality of life domains, and compare several summary utility measures. METHODS: In the ADVANCE (Action in Diabetes and Vascular Disease: Preterax and Diamicron MR Controlled Evaluation) study, 978 Australian patients with Type 2 diabetes completed two health-related quality of life questionnaires at baseline: the EQ-5D and the SF-36v2, from which nine summary utility measures were calculated, and compared. The algorithms were grouped into four classes: (i) based on the EQ-5D; (ii) using fewer items than those in the SF-12 (iii) using the items in the SF-12; and (iv) using all items of the SF-36. RESULTS: Overall health-related quality of life of the subjects was good (mean utility ranged from 0.68 (+/-0.08) to 0.85(+/-0.14) over the nine utility measures) and comparable to patients without diabetes. Summary indices were well correlated with each other (r = 0.76 to 0.99), and showed lower health related quality of life in patients with major diabetes-related events such as stroke or myocardial infarction. Despite the smaller number of items used in the scoring of the EQ-5D, it generally performed at least as well as SF-36 based methods. However, all utility measures had some limitation such as limited range or ceiling effects. CONCLUSION: The summary utility measures showed good agreement, and showed good discrimination between major and minor health state changes. However, EQ-5D based measures performed as well and are generally simpler to use. PMID- 17462101 TI - Blastocyst production by in vitro maturation and development of porcine oocytes in defined media following intracytoplasmic sperm injection. AB - The present study was carried out to establish porcine defined IVP. In Experiments 1 and 2, we investigated the efficacy of additional 0.6 mM cystine and/or 100 microM cysteamine (Cys) to a defined TCM199 maturation medium with regard to the intracellular glutathione (GSH) concentration and the developmental competence of in vitro matured porcine oocytes following intracytoplasmic sperm injection (ICSI). The control medium was a modified TCM199 containing 0.05% (w/v) polyvinyl alcohol (PVA). Cys and/or cystine were added to the control medium. The control group and immature oocytes (presumptive germinal vesicle oocytes; GV) were prepared for GSH assay. In Experiment 3, the efficacy of epidermal growth factor (EGF) addition to a modified porcine zygote medium (mPZM) for in vitro culture (IVC) medium was investigated on embryonic development and the mean cell number of blastocysts following ICSI. As a positive or negative control, 0.3% BSA (mPZM-3) or 0.3% PVA (mPZM-4), respectively, was added to the base medium. The defined IVC medium was supplemented with 5 or 10 ng/ml EGF. In Experiment 1, no significant difference was found in the rates of cleavage (31.4-64.3%) and blastocyst formation (6.5-22.9%) among the treatment and control groups. The mean cell numbers per blastocyst ranged from 30 to 48 among the groups without significant differences. However, in Experiment 2, the intracellular GSH concentrations in the oocytes cultured in the medium supplemented with 100 microM Cys (9.6 pmol/oocyte) or Cys + cystine (9.9 pmol/oocyte) were significantly (p < 0.05) higher than the control (2.5 pmol/oocyte) and 0.6 mM cystine (6.5 pmol/oocyte) groups, but not different from the GV group (9.0 pmol/oocyte). The GSH concentration in the cystine group was also significantly (p < 0.05) higher than that in the control group, but not different from the GV group. In Experiment 3, the rates of cleavage and blastocyst formation and the mean cell numbers of blastocysts were not significantly different among the groups. However, the addition of 5 ng/ml EGF into the mPZM-4 resulted in a significantly (p < 0.05) higher blastocyst rate per cleaved embryo than the other two defined groups (mPZM-4 + 5 ng/ml: 48.6%, mPZM-4 and mPZM-4 +10 ng/ml: 23.4% and 23.1%, respectively). The present results indicate that the addition of Cys to a defined medium for in vitro maturation (IVM) of porcine oocytes increases intracellular GSH concentration. Further addition of cystine into the IVM medium containing 100 microM Cys is not necessary and TCM199 plus Cys (100 microM) could be used as a defined IVM medium for porcine oocytes. The addition of 5 ng/ml EGF to a defined IVC medium has enhanced subsequent development after ICSI. This study shows that porcine blastocysts can be produced by defined media throughout the steps of IVP (IVM, ICSI and IVC). PMID- 17462102 TI - The ability of whale haploid spermatogenic cells to induce calcium oscillations and its relevance to oocyte activation. AB - Interspecies microinsemination assay was applied to examine the ability of minke whale haploid spermatogenic cells to induce Ca2+ oscillations and oocyte activation. Populations of round spermatids (RS), early-stage elongating spermatids (e-ES), late-stage elongating spermatids (1-ES) and testicular spermatozoa (TS) were cryopreserved in the presence of 7.5% glycerol on board ship in the Antarctic Ocean. Repetitive increases of intracellular Ca2+ concentration occurred in 0, 65, 81 and 96% of BDF1 mouse oocytes injected with the postthaw RS, e-ES, 1-ES and TS, respectively. A normal pattern of the Ca2+ oscillations was observed in 26-47% of the responding oocytes. Most oocytes that exhibited Ca2+ oscillations, regardless of the oscillation pattern, resumed meiosis (83-94%). These results indicate that whale spermatogenic cells acquire SOAF activity, which is closely related to their Ca2+ oscillation-inducing ability at the relatively early stage of spermiogenesis. PMID- 17462103 TI - Establishment of a basic method for manipulating preantral follicles: effects of retrieval method on in vitro growth of preantral follicles and intrafollicular oocytes. AB - The aim of this study was to establish a basic manipulation protocol of preantral follicles for deriving developmentally competent oocytes. Primary, early and late secondary follicles retrieved from the ovaries of 14-day-old F1 (C57BL/6 x DBA2) female mice mechanically or enzymatically were cultured singly and in vitro growth of the follicles and maturation of intrafollicular oocytes were subsequently monitored. A mechanical method retrieved more (p < 0.0001) follicles (339 +/- 48 vs. 202 +/- 28) than an enzymatic method. However, the enzymatic method collected more singly isolated follicles that could be provided for subsequent culture (102 +/- 26 vs. 202 +/- 28). When an enzymatic method was employed, early and late secondary follicles required 9 and 6 days for reaching the maximal incidence of the pseudoantral stage. However, primary follicles were not possible to develop into the pseudoantral stage. The optimal duration of oocyte maturation from the onset of follicle culture was 7 days and 5-7 days for early and late secondary follicles, respectively. A general decrease in oocyte diameter (65.2-65.53 microm vs. 75 microm) and zona thickness (5.41-5.74 microm vs. 7.76 microm) was detected in in vitro-derived compared with in vivo-derived matured oocytes. Pronuclear formation was detected in 86-94% of mature oocytes after parthenogenetic activation and no significant difference was detected among groups. These results showed that preantral follicles retrieved by an enzymatic method underwent step-by-step growth in vitro, which could yield mature oocytes. PMID- 17462104 TI - Analysis of transcription factor expression during oogenesis and preimplantation development in mice. AB - The transition from a differentiated germ cell into a totipotent zygote during oogenesis and preimplantation development is critical to the creation of a new organism. During this period, cell characteristics change dynamically, suggesting that a global alteration of gene expression patterns occurs, which is regulated by global changes in various epigenetic factors. Among these, transcription factors (TFs) are essential in the direct regulation of transcription and also play important roles in determining cell characteristics. However, no comprehensive analysis of TFs from germ cells to embryos had been undertaken. We used mRNA amplification systems and microarrays to conduct a genomewide analysis of TFs at various stages of oogenesis and preimplantation development. The greatest alteration in TFs occurred between the 1- and 2-cell stages, at which time zygotic genome activation (ZGA) occurs. Our analysis of TFs classified by structure and function revealed several specific patterns of change. Basic transcription factors, which are the general components of transcription, increased transiently at the 2-cell stage, while homeodomain (HD) TFs were expressed specifically in the oocyte. TFs containing the Rel homology region (RHR) and Ets domains were expressed at a high level in 2-cell and blastocyst embryos. Thus, the global TF dynamics that occur during oogenesis and preimplantation development seem to regulate the transition from germ-cell-type to embryo-type gene expression. PMID- 17462105 TI - Expression of SRG3, a chromatin-remodelling factor, in the mouse oocyte and early preimplantation embryos. AB - SRG3 (Smarcc1) is a core subunit of the SWI/SNF complex. In the absence of SRG3, embryonic development ceases during peri-implantation stages, indicating that SRG3, as well as the chromatin-remodelling process, plays an essential role in early mouse development. To gain a better understanding of chromatin remodelling during the early stages of development, we examined SRG3 expression during oogenesis and preimplantation stages using immunofluorescence and western blot assays. SRG3 was detected in nuclei of oocytes during growth and maturation. Following fertilization, SRG3 was detected in pronuclei shortly after their formation. Nuclear concentrations of SRG3 increased in a time-dependent fashion and were found to be greater in the male pronucleus than in the female pronucleus. The increase in nuclear SRG3 was partially inhibited by a protein synthesis inhibitor, but not by a transcriptional inhibitor. Expression of SRG3 is accompanied by expression of Brg1 and Ini1, two other core subunits of the SWI/SNF complex. The expression of these three remodelling factors parallels that of SP1 and TBP, both spatially and temporally, in the mouse embryo, suggesting a role for remodelling factors in chromatin structure and function during early development. PMID- 17462106 TI - Mitogen-activated protein kinase phosphorylation patterns in pig oocytes and cumulus cells during gonadotrophin-induced resumption of meiosis in vitro. AB - The present study investigated the phosphorylation pattern of mitogen-activated protein kinase (MAPK) in cumulus-oocyte complexes (COCs) during spontaneous and FSH/LH-induced in vitro maturation (IVM). Both isoforms of MAPK were unphosphorylated in oocytes recovered immediately after liberation from follicles and became phosphorylated following 25 h incubation, corresponding to the time of germinal vesicle breakdown (GVBD). In contrast, MAPK was already phosphorylated in minimal amounts in cumulus cells at the time of liberation from follicles and phosphorylation of MAPK increased after 0.5 h incubation. Supplementation of medium with gonadotrophins intensified phosphorylation at 0.5 h incubation, demonstrating the early and rapid action of FSH/LH on MAPK phosphorylation. Phosphorylation of MAPK in cumulus cells peaked after 21 h of incubation, whereas MAPK was almost completely dephosphorylated at the end of incubation (45 h). During subsequent incubation in the absence of added gonadotrophins, between 5 and 10 h exposure to FSH/LH-supplemented medium was required to induce resumption of meiosis in COCs. Phosphorylation of MAPK in oocytes was prevented by the MEK inhibitor U0126, but the inhibitor reduced phosphorylation of MAPK in cumulus cells only during the first 2 h of IVM. The data support the hypothesis that two different MAPK phosphorylation events occurred following gonadotrophin stimulation, one in cumulus cells and the other in oocytes. In cumulus cells, FSH/LH induced early and rapid U0126-insensitive phosphorylation of MAPK, whereas U0126-susceptible MAPK phosphorylation took place in the oocyte itself around the time of GVBD. PMID- 17462107 TI - Implication of gap junction coupling in amphibian vitellogenin uptake. AB - The aim of the present study was to investigate the physiological role and the expression pattern of heterologous gap junctions during Xenopus laevis vitellogenesis. Dye transfer experiments showed that there are functional gap junctions at the oocyte/follicle cell interface during the vitellogenic process and that octanol uncouples this intercellular communication. The incubation of vitellogenic oocytes in the presence of biotinylated bovine serum albumin (b-BSA) or fluorescein dextran (FDX), showed that oocytes develop stratum of newly formed yolk platelets. In octanol-treated follicles no sign of nascent yolk sphere formation was observed. Thus, experiments in which gap junctions were downregulated with octanol showed that coupled gap junctions are required for endocytic activity. RT-PCR analysis showed that the expression of connexin 43 (Cx43) was first evident at stage II of oogenesis and increased during the subsequent vitellogenic stages (III, IV and V), which would indicate that this Cx is related to the process that regulates yolk uptake. No expression changes were detected for Cx31 and Cx38 during vitellogenesis. Based on our results, we propose that direct gap junctional communication is a requirement for endocytic activity, as without the appropriate signal from surrounding epithelial cells X. laevis oocytes were unable to endocytose VTG. PMID- 17462108 TI - Microplate assay for quantifying developmental morphologies: effects of exogenous hyalin on sea urchin gastrulation. AB - It is often difficult to determine the effects of various substances on the development of the sea urchin embryo due to the lack of appropriate quantitative microassays. Here, a microplate assay has been developed for quantitatively evaluating the effects of substances, such as hyalin, on living sea urchin embryos. Hyalin (330 kDa) is a major constituent of the sea urchin hyaline layer, an extracellular matrix that develops 20 min postinsemination. Function of the hyaline layer and its major constituent, is the adhesion of cells during morphogenesis. Using wide-mouthed pipette tips, 25 microl of 24-h Strongylocentrotus purpuratus embryos were transferred to each well of a 96-well polystyrene flat-bottom microplate yielding about 12 embryos per well. Specific concentrations of purified hyalin diluted in low calcium seawater were added to the wells containing the embryos, which were then incubated for 24 h at 15 degree C. The hyalin-treated and control samples were observed live and after fixation with 10% formaldehyde using a Zeiss Axiolab photomicroscope. The small number of embryos in each well allowed quantification of the developmental effects of the added media. Specific archenteron morphologies-attached, unattached, no invagination and exogastrula-were scored and a dose-dependent response curve was generated. Hyalin at high concentrations blocked invagination. At low concentrations, it inhibited archenteron elongation/attachment to the blastocoel roof. While many studies have implicated hyalin in a variety of interactions during morphogenesis, we are not aware of any past studies that have quantitatively examined the effects of exogenous hyalin on specific gastrulation events in whole embryos. PMID- 17462109 TI - Effect of embryonic cell cycle of nuclear donor embryos on the efficiency of nuclear transfer in Japanese black cattle. AB - In the present study, the development in vitro and in vivo of nuclear transfer (NT) embryos reconstructed with embryonic cells (blastomeres) at the 32- to 63 cell (sixth cell cycle) and 64- to 127-cell (seventh cell cycle) stages was investigated to determine the optimum range of embryonic cell cycles for yielding the highest number of identical calves in Japanese black cattle. Rates of development to the blastocyst stage (overall efficiency) were higher in the sixth cell-cycle stage (45%) than in the seventh cell-cycle stage (12%). After the transfer of the blastocysts reconstructed with blastomeres of the sixth and seventh cell cycle-stage embryos to recipient heifers, there were no differences in the pregnancy (14/35: 40% versus 3/13: 23%, respectively) or calving rates (11/39: 28% versus 3/13: 23%, respectively). These results indicate that the highest number of identical calves would be obtained by using sixth cell cycle (32- to 63-cell)-stage embryos as nuclear donors. PMID- 17462110 TI - Essential role of follicle stimulating hormone in the maintenance of caprine preantral follicle viability in vitro. AB - The aims of the present study were to investigate the effects of follicle stimulating hormone (FSH) on survival, activation and growth of caprine primordial follicles using histological and ultrastructural studies. Pieces of caprine ovarian cortex were cultured for 1 or 7 days in minimum essential medium (MEM - control medium) supplemented with different concentrations of FSH (0, 10, 50 or 100 ng/ml). Small fragments from non-cultured ovarian tissue and from those cultured for 1 or 7 days in a specific medium were processed for classical histology and transmission electron microscopy (TEM). Additionally, effects of FSH on oocyte and follicle diameter of cultured follicles were evaluated. The results showed that the lowest percentage of normal follicles was observed after 7 days of culture in control medium. After 1 day of culture, a higher percentage of growing follicles was observed in the medium supplemented with 50 ng/ml of FSH. In the presence of 10 and 50 ng/ml of FSH, an increase in diameter of both oocyte and follicle on day 7 of culture was observed. TEM showed ultrastructural integrity of follicles after 1 day of culture in MEM and after 7 days in MEM plus 50 ng/ml FSH, but did not confirm the integrity of those follicles cultured for 7 days in MEM. In conclusion, this study demonstrated that FSH at concentration of 50 ng/ml not only maintains the morphological integrity of 7 days cultured caprine preantral follicles, but also stimulate the activation of primordial follicles and the growth of activated follicles. PMID- 17462111 TI - Effect of dehydroleucodine on meiosis reinitiation in Bufo arenarum denuded oocytes. AB - In amphibian oocytes meiosis, the transition from G2 to M phase is regulated by the maturation promoting factor (MPF), a complex of the cyclin-dependent kinase p34/cdc2 and cyclin B. In immature oocytes there is an inactive complex (pre MPF), in which cdc2 is phosphorylated on both Thr-161 and Thr-14/Tyr-15 residues. The dephosphorylation of Thr-14/Tyr-15 is necessary for the start of MPF activation and it is induced by the activation of cdc25 phosphatase. Late, to complete the activation, a small amount of active MPF induces an auto amplification loop of MPF stimulation (MPF amplification). Dehydroleucodine (DhL) is a sesquiterpenic lactone that inhibits mammalian cell proliferation in G2. We asked whether DhL interferes with MPF activation. For this question, the effect of DhL (up to 30 microM) on the resumption of meiosis was evaluated, and visualized by germinal vesicle break down (GVBD), of Bufo arenarum oocytes induced in vitro by either: (i) removing follicle cells; (ii) progesterone stimulation; (iii) VG-content injection; or (iv) injection of mature cytoplasm. The results show that DhL induced GVBD inhibition, in a dose-dependent manner, in spontaneous and progesterone-induced oocyte maturation. Nevertheless, DhL at the doses assayed had no effect on GVBD induced by mature cytoplasm injection, but exerted an inhibitory effect on GVBD induced by GV content. On the basis of these results, we interpreted that DhL does not inhibit MPF amplification and that the target of DhL is any event in the early stages of the cdc25 activation cascade. PMID- 17462112 TI - Immunology of pregnancy: cellular mechanisms allowing fetal survival within the maternal uterus. AB - Pregnancy success remains a fascinating phenomenon to immunologists as it defies the immunological rules of rejection. Although it was previously thought that the maternal immune system does not see the fetus, it is now well documented that fetal cells reach the maternal body and encounter host immune cells. Natural tolerance mechanisms following this interaction remain to be fully elucidated. This article reviews the current literature on mechanisms of adaptive immunity, with emphasis on regulatory T cells and heme oxygenase 1 (HO-1). We propose a scenario in which regulatory T cells create a tolerant microenvironment at the fetal-maternal interface characterised by the presence of tolerance-associated molecules such as HO-1, which has been shown to be of vital importance for fetal survival. PMID- 17462114 TI - End-of-life patients, intensive care and consent: difficulties facing French intensivists. AB - BACKGROUND: The French legislature passed a law in 2005 that assigns a new role to the physician and redefines his liability in end-of-life decisions. METHOD: This law is presented and discussed in context with current French legal practice. RESULTS: This law emphasizes patient autonomy, advocating that the patient be fully informed before treatment, and creates specific procedures to be followed according to whether the patient is conscious or unconscious. In the latter situation, the law reinforces the role of both the patient's surrogate and the patient's advance directives in establishing consent. In these extreme situations, doctors have the option to request a second medical opinion. This joint decision-making procedure is laid down by law and becomes obligatory in the interests of transparency. CONCLUSION: Respect for patients' consent implies the possibility that they may refuse medical care, creating an ethical and legal dilemma of providing medical care or respecting the patients' wishes. The key issue concerning end-of-life patients rests in the decisions taken concerning the continuation or withdrawal of life support and the administration of palliative care. PMID- 17462115 TI - The pharmacodynamics of mivacurium in the rabbit with carbon tetrachloride induced liver disease. AB - BACKGROUND AND OBJECTIVE: Neuromuscular blocking effects according to the severity of liver dysfunction have not been evaluated. We assessed the neuromuscular effects of mivacurium in carbon tetrachloride (CCl4)-treated rabbits with toxic hepatitis in vivo. METHODS: We compared the dose-response relationships and the neuromuscular blocking effects of mivacurium in 66 rabbits randomly treated with 0.3 mL kg(-1) of corn oil, 0.3 mL kg(-1) of CCl4 or 0.6 mL kg(-1) of CCl4 for 11 weeks, respectively. Train-of-four stimuli were applied every 10 s to the common peroneal nerve and the force of contraction of the tibialis anterior muscle was measured. RESULTS: Severe hepatitis was associated with a rightward shift of the mivacurium dose-response curves, but mild hepatitis had no effect. The calculated ED50 values for the control, mild and severe hepatitis were 17.1+/-2.6, 18.2+/-2.7 and 31.8+/-3.2 microg kg(-1), respectively. Rabbits with severe hepatitis had a significantly prolonged recovery time from neuromuscular blockade compared with other rabbits. Cholinesterase activity had a negative correlation with recovery indices of mivacurium even in severe hepatic injury. Severe hepatitis induced a prolongation of action duration of repeated doses, but maintained the constant intervals. CONCLUSIONS: The dose-response and the time course of neuromuscular blockade of mivacurium differ in mild hepatitis compared with severe hepatitis, but required no adjustments of different doses for repeated injection after the desired depth of neuromuscular block, and had a negative correlation with the activity of plasma cholinesterase in both hepatic injuries. PMID- 17462116 TI - Perioperative feasibility of imaging the heart and pleura in patients with aortic stenosis undergoing aortic valve replacement. AB - BACKGROUND: The postoperative monitoring and treatment of the patient undergoing aortic valve replacement is a complex challenge. Echocardiography is the only method which provides dynamic and real-time bedside imaging of the heart. Focused assessed transthoracic echocardiography has been shown to provide a usable window for cardiac imaging in a mixed ICU population. The aim of this study was to evaluate the feasibility of perioperative imaging of the heart and pleura according to the focused assessed transthoracic echocardiography protocol in patients scheduled for aortic valve replacement. METHOD: Thirty-five adult patients scheduled for aortic valve replacement were followed perioperatively with focused assessed transthoracic echocardiography examinations. A Vivid-7 echo machine and a 2.5 MHz matrix transducer with second-harmonic imaging were used for data acquisition. The image quality for the cardiac window was graded 1-5 (1 = no image, 2 = poor and unusable image quality, 3 = usable image quality, 4 = good image quality and 5 = perfect image quality). A score >or=3 equalled an image quality judged to be of sufficient quality to be interpreted and thereby to contribute to clinical decision-making. RESULTS: All patients had at least one usable window preoperatively. At least one usable window was obtained in 88% of patients on the first postoperative day, and in 97% at discharge. The image quality changed over time, with the poorest quality being observed on the first postoperative day. The apical view with the patient in the left lateral position provided the best cardiac window on all occasions. The presence of drains did not significantly affect the achievability of a satisfactory examination. The number of patients with pleural effusion was relatively high. On the first postoperative day, 10 subjects had unilateral and one subject had bilateral pleural effusions. At discharge, 14 patients had unilateral and four patients had bilateral effusions. CONCLUSIONS: We conclude that the image quality of the heart and pleura, according to the focused assessed transthoracic echocardiography concept, is sufficient to undergo interpretation and thereby contribute to the perioperative clinical decision-making in patients with aortic stenosis. PMID- 17462117 TI - Auditory-evoked potentials in bispectral index-guided anaesthesia for cardiac surgery. AB - BACKGROUND AND OBJECTIVE: Midlatency auditory-evoked potentials, as measures of the anaesthetic state, were evaluated at similar levels of bispectral index in cardiac surgical patients maintained with either propofol or isoflurane anaesthesia. METHODS: Twenty-four patients were randomly allocated to anaesthesia with propofol (n = 12) or isoflurane (n = 12). Bispectral index was maintained below 60 during surgery. Auditory-evoked potentials were collected before induction of anaesthesia, 10 min after intubation, 30 min after sternotomy, during cardiopulmonary bypass at the time of cross-clamping of the aorta and during stable mild hypothermia, after de-clamping of the aorta, and after the operation. RESULTS: At the pre-determined time points, bispectral index values showed comparable depth of hypnosis in both groups. The latency of the Nb component of midlatency auditory-evoked potentials was significantly increased in the isoflurane group after intubation (P < 0.001) and that of both the Nb and the Pa components after sternotomy (P < 0.001) compared with the propofol group. No differences between the groups were detected with respect to haemodynamic variables. No patient reported recall of intraoperative events. CONCLUSION: After intubation and surgical stimulation, when bispectral index was at a constant level, there was a difference in the Nb and Pa components of the midlatency auditory-evoked potentials between the two anaesthetic regimens, indicating a distinction in the state of anaesthesia. Our results suggest that the parallel use of these two electrophysiological methods can show differences in the components of anaesthesia between various anaesthesia methods in cardiac surgical patients. PMID- 17462118 TI - Effects of ifenprodil on voltage-gated tetrodotoxin-resistant Na+ channels in rat sensory neurons. AB - BACKGROUND AND OBJECTIVE: To examine a possible mechanism for the antinociceptive action of the N-methyl-D-aspartate receptor antagonist ifenprodil, we compared its effects with those of ketamine on tetrodotoxin-resistant Na+ channels in rat dorsal root ganglion neurons, which play an important role in the nociceptive pain pathway. METHODS: Experiments were performed on dorsal root ganglion neurons from Sprague-Dawley rats, recordings of whole-cell membrane currents being made using patch-clamp technique. RESULTS: Both drugs blocked tetrodotoxin-resistant Na+ currents dose dependently, their half-maximal inhibitory concentrations being 145+/-12.1 micromol (ketamine) and 2.6+/-0.95 micromol (ifenprodil). Ifenprodil shifted the inactivation curve for tetrodotoxin-resistant Na+ channels in the hyperpolarizing direction and shifted the activation curve in the depolarizing direction. Use-dependent blockade of tetrodotoxin-resistant Na+ channels was more marked with ifenprodil than with ketamine. When paired with lidocaine, these drugs produced similar additive inhibitions of tetrodotoxin-resistant Na+ channel activity. CONCLUSIONS: The observed suppressive effects on tetrodotoxin-resistant Na+ channel activity may, at least in part, underlie the antinociceptive effects of these N-methyl-D-aspartate receptor antagonists. PMID- 17462119 TI - Regular tramadol use does not affect the propofol dose requirement for induction of anaesthesia. AB - BACKGROUND AND OBJECTIVES: An increased risk of awareness during general anaesthesia in patients receiving tramadol has been reported. We studied whether tramadol affects the amount of propofol required for induction of anaesthesia. METHODS: In this prospective controlled study, we evaluated 46 patients, half of whom used tramadol regularly. Entropy indices, state entropy and response entropy, were used to assess the level of hypnosis. Patients were anaesthetized with a propofol infusion (1 mg kg(-1) min(-1)) until they first became unconscious, and further until they developed a burst suppression pattern in the electroencephalogram. The doses of propofol needed to reach these end-points were recorded. RESULTS: The amount (median, (range)) of propofol required for loss of consciousness was 2.0 (1.0-5.5) mg kg(-1) and 2.4 (0.9-8.3) mg kg(-1) (P=0.95) in the tramadol users and controls, respectively. The amount of propofol required for burst suppression was 5.8 (3.9-12.7) mg kg(-1) and 6.4 (2.9-15.1) mg kg(-1) (P=0.89) in the tramadol users and controls. There was no difference between the groups in state entropy and response entropy during different stages of induction of anaesthesia. CONCLUSIONS: Tramadol did not affect the dose of propofol required to achieve loss of consciousness or burst suppression pattern in electroencephalogram during induction of general anaesthesia. However, there was a ninefold inter-individual variation in propofol dose requirement for loss of consciousness and a fivefold variation for reaching burst suppression. Due to extensive inter-individual variability, monitoring the level of hypnosis during general anaesthesia using propofol may enhance the correct dosage. PMID- 17462120 TI - Biphasic cardiac output changes during onset of spinal anaesthesia in elderly patients. AB - BACKGROUND AND OBJECTIVE: In most studies of cardiac output changes after spinal anaesthesia, the time-resolution is limited. The aim of this study was to demonstrate cardiac output changes with high time-resolution during onset of spinal anaesthesia in elderly patients. METHODS: We investigated 32 patients aged 60 yr scheduled for elective lower limb surgery. Fourteen received concurrent cardiovascular medication. Cardiac output was measured every 10 s using a pulse wave algorithm derived from the radial artery pressure curve, after calibration with lithium chloride (LiDCOplus). Data collection ended when the patients were ready for surgery, or if ephedrine was given to raise the mean arterial pressure. RESULTS: Cardiac output increased initially reaching a maximum after a mean of 7 min. The average increase was 1.1 L min(-1) (P<0.0001). This occurred when mean arterial pressure was reduced 14 mmHg on average. At the end of data collection, cardiac output decreased 0.5 L min(-1) from baseline (P=0.02). Mean arterial pressure decreased progressively in all patients, and only minimal changes in heart rate were found. CONCLUSIONS: Using this high time-resolution method, we detected biphasic changes in cardiac output during onset of spinal anaesthesia. Initially, cardiac output increased. Subsequently, it was significantly reduced from baseline, although this decrease was of minor clinical importance. PMID- 17462121 TI - Establishment of the onset of host specificity in four phyllobothriid tapeworm species (Cestoda: Tetraphyllidea) using a molecular approach. AB - A parasitological survey in the Bay of Fundy, Canada, resulted in the recovery of mature specimens from 5 species of phyllobothriid tapeworms (Cestoda: Tetraphyllidea) from 4 rajid skates: Echeneibothrium canadensis and E. dubium abyssorum specimens from Amblyraja radiata; E. vernetae and Pseudanthobothrium n.sp. from Leucoraja erinacea and L. ocellata; and P. hanseni from A. radiata and Malacoraja senta. Partial sequence data of a variable region (D2) from the large subunit ribosomal DNA (LSU) were used here to determine the host distribution of immature specimens for 4 of these 5 species (E. d. abyssorum was not included in the analyses). Immature specimens from both Pseudanthobothrium spp. were identified in the same hosts as recorded previously for mature specimens, thus suggesting that there are mechanisms that prevent the attachment of the parasite in an 'unsuitable' host species. Immature E. canadensis specimens were recovered exclusively from A. radiata, whereas immature E. vernetae specimens were recovered from L. erinacea and A. radiata, despite the latter host species not harbouring mature E. vernetae specimens. Their presence in the latter host species may be explained by host restriction or resistance, which allows the attachment of the parasites in the 'wrong' host species, but not establishment or development. PMID- 17462122 TI - Molecular phylogenetic analysis in Hammondia-like organisms based on partial Hsp70 coding sequences. AB - The 70 kDa heat-shock protein (Hsp70) sequences are considered one of the most conserved proteins in all domains of life from Archaea to eukaryotes. Hammondia heydorni, H. hammondi, Toxoplasma gondii, Neospora hughesi and N. caninum (Hammondia-like organisms) are closely related tissue cyst-forming coccidians that belong to the subfamily Toxoplasmatinae. The phylogenetic reconstruction using cytoplasmic Hsp70 coding genes of Hammondia-like organisms revealed the genetic sequences of T. gondii, Neospora spp. and H. heydorni to possess similar levels of evolutionary distance. In addition, at least 2 distinct genetic groups could be recognized among the H. heydorni isolates. Such results are in agreement with those obtained with internal transcribed spacer-1 rDNA (ITS-1) sequences. In order to compare the nucleotide diversity among different taxonomic levels within Apicomplexa, Hsp70 coding sequences of the following apicomplexan organisms were included in this study: Cryptosporidium, Theileria, Babesia, Plasmodium and Cyclospora. Such analysis revealed the Hammondia-like organism to be the lowest divergent group when compared to other groups within the phylum Apicomplexa. In conclusion, the Hsp70 coding sequences proved to be a valuable genetic marker for phylogenetic reconstruction and may constitute a good candidate to be used with other genes for species phylogeny within this group of organisms. PMID- 17462123 TI - Genetic variation in the mitochondrial cytochrome c oxidase subunit 1 within Progamotaenia festiva (Cestoda: Anoplocephalidae) from macropodid marsupials. AB - Genetic variation was examined in the anoplocephalid cestode Progamotaenia festiva, from Australian marsupials, in order to test the hypothesis that P. festiva, is a complex of sibling species and to assess the extent of host switching reported previously based on multilocus enzyme electrophoresis (MEE). Polymerase chain reaction (PCR)-based single-strand conformational polymorphism (SSCP) was used for the analysis of sequence variation in the cytochrome c oxidase subunit 1 (cox1) gene among 179 specimens of P. festiva (identified based on morphology and predilection site in the host) from 13 different host species, followed by selective DNA sequencing. Fifty-three distinct sequence types (haplotypes) representing all specimens were defined. Phylogenetic analyses of these sequence data (utilizing maximum parsimony and neighbour-joining methods) revealed 12 distinct clades. Other heterologous species, P. ewersi and P. macropodis, were used as outgroups and the remaining bile-duct inhabiting species, P. diaphana and P. effigia, were included in the analysis for comparative purposes. The latter 2 species were nested within the clades representing P. festiva. Most clades of P. festiva identified were restricted to a single host species; one clade primarily in Macropus robustus was also found in the related host species M. antilopinus in an area of host sympatry; another clade occurring primarily in M. robustus occurred also in additional kangaroo species, M. rufus and M. dorsalis. High levels of genetic divergence, the existence of distinct clades and their occurrence in sympatry provide support for the hypothesis that P. festiva represents a complex of numerous species, most of which, but not all, are host specific. Three distinct clades of cestodes were found within a single host, M. robustus, but there was no evidence of within-host speciation. PMID- 17462124 TI - Novel approach to performing metabolite identification in drug metabolism. AB - A novel online method is developed, using liquid chromatography (LC)-accurate radioisotope counting dynamic-flow (ARC) coupled with a radioactivity detector and mass spectrometer, for metabolite identification in drug discovery and development. This method offers the advantages of improved sensitivity for detecting radiolabeled drugs as well as streamlining the process of identifying and characterizing metabolites. For the purposes of evaluating this method, in vitro human liver microsomal incubations with [(14)C]dextromethorphan are conducted. Online separation and identification of [(14)C]dextromethorphan metabolites are achieved without intensive sample preparation, concentration, or fraction collection. Mass spectrometric analysis identified and characterized the metabolites of dextromethorphan formed by N - and O -dealkylation, correlated well with previously published results. Chromatographic peaks for [(14)C]dextromethorphan and its metabolites are collected online, then infused for extended periods of time at a flow rate of 10 microL/min while maintaining the column pressure. The continuous analytical signal input allowed acquisition of a higher order of multistage fragmentation for both major and minor metabolites. The multistage MS fragmentation pattern obtained for the metabolites allowed defining the sites of metabolism for dextromethorphan. Further evaluations of this method are also conducted using a [(14)C]compound A to check the linearity and sensitivity of the dynamic-flow method. The R(2) value is 0.996 for the dynamic-flow method between 50 and 600 disintegrations per minute (dpm); the limit of detection for LC-ARC is 20 dpm, which is approximately 10 times more sensitive than conventional continuous-flow radioactivity detection techniques. The overall results suggest that the combination of LC-ARC with radioactivity detection and mass spectrometry has great potential as a powerful tool for enhancing the sensitivity of radioisotope measurement in metabolite identification studies during drug discovery and development. PMID- 17462125 TI - Monitoring by liquid chromatography coupled to mass spectrometry the impact of pH and temperature on the pigment pattern of cactus pear fruit extracts. AB - The influence of pH and moderate heating (50 degrees C) on the color and individual betacyanin content of pigment extracts from cactus pear fruits (Opuntia stricta) is studied in the course of this paper. The study is carried out by using a high-performance liquid chromatograph equipped with a photodiode array detector and coupled to a mass spectrometer. The results point to a pH dependent degradation mechanism, which is reflected in the chromatographic patterns obtained at different exposure times (0-28 h). At pH 3, 15-descarboxy betanin is the most resistant betacyanin derivative. At pH 5, seven peaks are detected after 8 h, the most prominent being betanin, cyclo-dopa-5-O-beta glucoside, and betalamic. In the assay conducted at pH 7, rapid color loss affects all the pigments, except for betanin. PMID- 17462126 TI - Analysis of monoethanolamine by derivatization with Marfey's reagent and HPLC. AB - A sensitive and selective method for determining the residual monoethanolamine in a developmental drug substance is developed and validated. Marfey's reagent, which is commonly used for the chiral analysis of amino acids, is reacted with the primary amine group of monoethanolamine and then analyzed by high-performance liquid chromatography-UV at 340 nm. Quantitation is performed by a standard addition method by preparing drug substance samples with added monoethanolamine ranging from 0.25-1.0 microg/mL (equivalent to 12.5-50 ppm with respect to the drug substance). The method performance is evaluated for linearity, specificity, detection and quantitation limits, accuracy, precision, and sample stability. The method is linear from 0.25-1.0 microg/mL with a coefficient of determination (r(2)) > 0.95. The accuracy and precision obtained is 105.5 +/- 4.8% (n = 3). The limits of detection and quantitation are 0.03 and 0.10 microg/mL, respectively. Instrument precision (% relative standard deviation of six injections of a derivatized 0.5 microg/mL monoethanolamine solution on two separate days) is >/= 2.0%. This method is suitable for the determination of monoethanolamine at the 25 ppm level in drug substance. PMID- 17462127 TI - Analysis of some chlorophenoxy acids and carbamate herbicides in water and soil as amide derivatives using gas chromatography-mass spectrometry. AB - A newly developed method for determining three phenoxy acids and one carbamate herbicide in water and soil samples using gas chromatography with mass spectrometric detection is developed. Phenoxy acids are derivatized through a condensation reaction with a suitable aromatic amine. 1,1-Carbonyldiimidazole is used as a condensation reagent. Derivatization conditions are optimized with respect to the amount of analyte, amine, solvent, and derivatization reagent. The optimum derivatization yield is accomplished in acetonitrile. 4-Methoxy aniline is used as a derivatizing agent. Obtained derivatives are stable indefinitely. Enhancement in sensitivity is achieved by using the single-ion monitoring mass spectrometric mode. The effectiveness of the developed method is tested by determining investigated compounds in water and soil samples. Analytes are concentrated from water samples using liquid-phase extraction and solid-phase extraction. Soil samples are extracted using methanol. Detection limits of 1.00, 50.00, 100.00, and 1.00 ng/mL are obtained for 2-(1-methylethoxy)phenyl methylcarbamate (Baygon), 2-(3-chlorophenoxy)-propionic acid (Cloprop), 2,4,5 trichlorophenoxyacetic acid, and 4-(2,4-dichlorophenoxy)butyric acid, respectively. LPE for spiked water samples yields recoveries in the range of 60.6 95.7%, with relative standard deviation (RSD) values of 1.07-7.85% using single component calibration curves. Recoveries of 44.8-275.5%, with RSD values ranging from 1.43% to 8.61% were obtained using a mixed component calibration curves. SPE from water samples and soil samples showed low recoveries. The reason is attributed to the weak sorption capabilities of soil and Al(2)O(3). PMID- 17462128 TI - Study of retention parameters obtained in RP-TLC system and their application on QSAR/QSPR of some alpha adrenergic and imidazoline receptor ligands. AB - The retention constant (R(0)(m)) is determined for 11 selected adrenergic and imidazoline receptor ligands by reverse-phase-thin layer chromatography. It is established that the retention behavior of investigated compounds mostly depends on geometrical, electrostatic, and hydrogen bonding properties. Good correlations among hydrophobic parameters R(0)(m) versus log P for all eleven tested compounds are obtained. The satisfactory correlations are found between R(0)(m) versus apparent partition coefficient octanol-buffer pH 7.4 (log P') or apparent partition coefficient in four liposome systems (log K'(M)) and hypotensive activity (pC(25)) for five imidazolines. The results confirm the suitability of this parameter in quantitative structure-property and structure-activity relationships studies of these drugs. PMID- 17462129 TI - On-line SPE-Nano-LC-Nanospray-MS for rapid and sensitive determination of perfluorooctanoic acid and perfluorooctane sulfonate in river water. AB - An instrumental set up including on-line solid-phase extraction, nano-liquid chromatography, and nanospray mass spectrometry is constructed to improve the sensitivity for quantitation of perfluorooctanoic acid (PFOA) and perfluorooctane sulfonate (PFOS) in surface water. Sample volumes of 1000 microL are loaded onto a microbore 1.0-mm i.d. x 5 mm, 5 microm Kromasil C(18) enrichment column by a carrier solution consisting of 10mM ammonium acetate in acetonitrile-water (10:90, v/v) at a flow rate of 250 microL/min, providing on-line analyte enrichment and sample clean-up. Backflushed elution onto a 0.1-mm i.d. x 150 mm, 3.5 microm Kromasil C(18) analytical column is conducted using an acetonitrile 10mM ammonium acetate solvent gradient from 30% to 70% acetonitrile. Water samples are added with internal standard (perfluoroheptanoic acid) and filtrated prior to injection. The mass limits of detection of PFOA and PFOS are 0.5 and 1 pg, respectively, corresponding to concentration limits of detection of 500 pg/L and 1 ng/L, respectively. The total time spent on sample preparation, chromatography, and detection is approximately 12 min per sample. The method was employed for the determination of PFOS and PFOA in urban river water. PMID- 17462130 TI - Gas chromatography method for the characterization of ethanol steam reforming products. AB - Ethanol steam reforming is a promising reaction for producing fuel cell hydrogen. Depending on catalyst and reaction conditions, mixtures of condensable hydrocarbons and organic and inorganic gases are produced. This paper proposes an economic and effective solution for separating and detecting these compounds employing a gas chromatograph equipped with two columns, two 6-way valves, and two detectors. PMID- 17462131 TI - High-performance liquid chromatography-electrospray ionization-mass spectrometric determination of emedastine difumarate in human plasma and its pharmacokinetics. AB - A selective and sensitive method employing high-performance liquid chromatography (HPLC)-electrospray ionization (ESI)-mass spectrometry is developed and validated for the determination of emedastine difumarate in human plasma. With naphazoline hydrochloride as the internal standard, emedastine difumarate is extracted from plasma with ethyl acetate. The organic layer is evaporated, and the residue is redissolved in the mobile phase. An aliquot of 10 microL is chromatographically analyzed on a prepacked Phenomenex Luna 5u CN 100A (150 x 2.0-mm i.d.) column, using a mobile phase comprised of methanol-water (20 mM CH(3)COONH(4), pH 4.0) (80:20, v/v). Standard curves are linear (r(2) = 0.9990) over the concentration range of 0.05-30 ng/mL and had good accuracy and precision. The within- and between-batch precisions did not exceed 15% for the relative standard deviation. The lower limit of detection is 0.01 ng/mL. The validated HPLC-ESI-MS method is successfully used to study emedastine difumarate pharmacokinetics in 12 healthy volunteers. PMID- 17462132 TI - A sound foundation: new-style dental postgraduate training. PMID- 17462133 TI - Infection control: a survey of UK special care dentists and dental care professionals. AB - AIM: To determine attitudes to and practice of infection control among practitioners involved in special care dentistry. METHODS: A questionnaire on the issues of decontamination and infection control was sent to all 680 members of the British Society for Disability and Oral Health. RESULTS: The response rate was 63.5%. Almost all respondents (95%) reported having a local infection-control policy; most (97%) had local protocols for the management of inoculation injuries; most (81%) gave new staff training in infection-control procedures and most (74%) provided updates for established staff. Most respondents 'usually' or 'always' provided eye protection for patients (95%) and themselves (93%). Virtually all routinely wore gloves, and nearly all (94%) 'always' changed gloves between patients. The majority 'usually' or 'always' disinfected or disposed of surface coverings between patients (98%), sterilised all non-disposable instruments that had been set out for the patient (99%), and disinfected laboratory work (96%). CONCLUSIONS: There was high awareness of infection-control issues, and good reported compliance among these dental workers. PMID- 17462135 TI - Primary double tooth: report of a rare correlation with the permanent dentition. AB - Double tooth is a developmental anomaly that is seen more frequently in the primary than the permanent dentition. Double tooth involving primary canines and lateral incisors is found only in the mandible. The succeeding permanent lateral incisor is often found missing congenitally in the same region. This article reports a rare case of a double tooth involving a mandibular primary canine and lateral incisor. Instead of agenesis, the double tooth has been succeeded by a permanent supernumerary tooth in the same region. PMID- 17462137 TI - Reply letters following orthodontic consultations: an audit of Merseyside general dental practitioners' satisfaction. AB - OBJECTIVES: To assess the opinions of general dental practitioners (GDPs) working within the National Health Ser vice (NHS) in Merseyside regarding the length, format and appropriateness of consultant orthodontists' letters sent in reply to referral letters. MAIN OUTCOME MEASURES: These were GDPs' satisfaction with the length, format and content of the reply letters; GDPs' preferences for the information deemed necessary in such letters; GDPs' awareness of the status of their patients and the actions they were requested to undertake. METHOD: After piloting, questionnaires were mailed to 330 participating GDPs between August and October 2004. RESULTS: Two hundred and fifty-one (76%) questionnaires were returned. With one exception, the length of consultants' letters was thought to be adequate. Of the responding GDPs 82% stated a preference for summaries of the diagnosis and treatment plan to be presented in a list format rather than as free text. Aspects of the treatment plan were thought to be more important than the information about the examination. Following receipt of the reply letter only 71% of GDPs were aware of what was happening to their patients and 75% knew what actions they were to undertake. CONCLUSIONS: This audit provides evidence that GDPs consider that a significant portion of the information in consultants' reply letters is not needed. Relevant information may be poorly communicated. PMID- 17462139 TI - Cardiovascular and oral disease interactions: what is the evidence? AB - This paper reviews the evidence for the interaction of oral disease (more specifically, periodontal infections) with cardiovascular disease. Cardiovascular disease is a major cause of death worldwide, with atherosclerosis as the underlying aetiology in the vast majority of cases. The importance of the role of infection and inflammation in atherosclerosis is now widely accepted, and there has been increasing awareness that immune responses are central to atherogenesis. Chronic inflammatory periodontal diseases are among the most common chronic infections, and a number of studies have shown an association between periodontal disease and an increased risk of stroke and coronary heart disease. Although it is recognised that large-scale intervention studies are required, pathogenic mechanism studies are nevertheless required so as to establish the biological rationale. In this context, a number of hypotheses have been put forward; these include common susceptibility, inflammation via increased circulating cytokines and inflammatory mediators, direct infection of the blood vessels, and the possibility of cross-reactivity or molecular mimicry between bacterial and self antigens. In this latter hypothesis, the progression of atherosclerosis can be explained in terms of the immune response to bacterial heat shock proteins (HSPs). Because the immune system may not be able to differentiate between self HSP and bacterial HSP, an immune response generated by the host directed at pathogenic HSP may result in an autoimmune response to similar sequences in the host. Furthermore, endothelial cells express HSPs in atherosclerosis, and cross reactive T cells exist in the arteries and peripheral blood of patients with atherosclerosis. Each of these hypotheses is reviewed in light of current research. It is concluded that although atherosclerotic cardiovascular disease is almost certainly a multifactorial disease, there is now strong evidence that infection and inflammation are important risk factors. As the oral cavity is one potential source of infection, it is wise to try to ensure that any oral disease is minimised. This may be of significant benefit to cardiovascular health and enables members of the oral health team to contribute to their patients' general health. PMID- 17462140 TI - Removable prostheses revisited: challenges for primary dental care. I. Complete dentures. AB - The provision of removable prostheses remains a significant aspect of primary dental care. In the recent past and in the near future, there have been and will be significant new influences on how removable prostheses are provided for patients in the UK. In this paper, the first of a series of three, the trends in edentulousness in the UK will be summarised, and important changes that are likely to influence complete denture provision will be outlined. The paper will then go on to provide an update of clinical procedures in complete denture provision. A second paper will consider partial denture provision, and a third paper will deal with overdentures. PMID- 17462142 TI - A longitudinal study into changing ethical attitudes of newly-qualified dentists in South-East England. AB - OBJECTIVE: To report the ethical stances taken by newly-qualified dental graduates working in the south-east of England and how these changed over their first two years in practice. METHODS: A longitudinal self-completed questionnaire based study using scenarios and closed questions was completed by an initial group of 135 vocational dental practitioners at the very beginning of their vocational training year, at the end of the year, and, finally, at the end of the subsequent year. Their answers were then analysed to see whether or not their views had changed during this period. RESULTS: At the commencement of the study in 2003, 133 (99%) newly-qualified dental graduates completed the questionnaire and answered the closed questions. The following year, 129 (96%) completed these documents and in 2005, at the end of the study, this figure was 97 (72%). There was considerable variation in the attitudes taken by the responding dentists. Reported attitudes changed over the study period in seven of the nine scenarios. Responses to the scenarios used could be grouped into three categories: those in which there was no change; those in which the change happened following completion of vocational training; and those in which there was a continual change. Answers to the four closed questions indicated a growing negativity to practice within the National Health Service over the study period. CONCLUSIONS: Dental educators need to be aware of the findings from this study. The importance of the ethical attitudes held and the reasons underpinning them should be explored if problems with delivery of care are to be avoided in the future. PMID- 17462145 TI - [New concept on internal fixation for maxillofacial bone fracture]. PMID- 17462146 TI - [Influence of prior radiotherapy on free tissue transfer in head and neck region]. AB - OBJECTIVE: To analyze the influence of prior radiotherapy on free tissue transfer in head and neck region. METHODS: One thousand and sixty-six consecutive free flap transfers performed in 1007 patients in head and neck region from May 1999 to March 2005 were reviewed. Among the 1007 cases, 146 had a history of prior radiotherapy (radiation group), and the other 896 without prior radiotherapy (non radiation group). The two groups were compared in terms of free flap success rate and postoperative complication rate. RESULTS: The success rate of free flap was 97.6% in the radiation group, and 98.4% in the non-radiation group. The postoperative complication rate was 38.4% in the radiation group, and 36.4% in the non-radiation group. CONCLUSIONS: Prior radiotherapy poses no significant influence on free tissue transfer in head and neck region. PMID- 17462147 TI - [The characteristics and significance of microvessel in oral squamous cell carcinoma]. AB - OBJECTIVE: To investigate the characteristics of distribution status of maturation and morphology of the microvessel in oral squamous cell carcinoma (OSCC) and analyze the correlations between the characteristics and clinicopathologic parameters. METHODS: The microvessel in 62 cases of OSCC, 30 cases of adjacent normal tissue and 10 cases of normal oral mucosa were detected by double-labeling immunohistochemistry technique. Image analysis mucosa, the microvascular density (MVD) was higher while vessel maturation system was also used for the analysis RESULTS: Compared with adjacent normal tissues and normal oral index (VMI), vascular form factor (VFF) and vascular perimeter (VP) were lower in OSCC with significant difference (P < 0.01). The MVD was higher (P < 0.01) and VMI was lower (P < 0.05) in peritumoral site than those in intratumoral site. The MVD were correlated significantly with lymph node metastasis (P < 0.01). CONCLUSIONS: The microvessels in OSCC were significantly different from those in normal oral tissues. The differences may contribute to the application of antiangiogenic therapy, and the peritumoral area in OSCC may become a target site of the therapy. The MVD may be hopefully used as an indicator of tumor malignancy. PMID- 17462148 TI - [Study on the inhibition of vascular endothelial growth factor expression in Tca8113 cell by inducible nitric oxide synthase gene RNA interference]. AB - OBJECTIVE: To investigate the effect of inducible nitric oxide synthase (iNOS) on vascular endothelial growth factor (VEGF) expression. METHODS: The vector containing short hairpin RNA of iNOS was transfected into Tca8113 cells using the RNA interference (RNAi) technique. The gene and protein expression of iNOS and VEGF was examined by RT-PCR and Western blot. RESULTS: The expression of iNOS, VEGF gene in Tca8113 cells was significantly different between the experimental and control groups 24 h and 48 h after transfection (P < 0.05). The protein expression of iNOS was different between the two groups 36 h and 48 h after transfection, and of VEGF was also different between the two groups 48 h after transfection (P < 0.05). CONCLUSIONS: The expression of VEGF could be down regulated by silencing the iNOS gene in Tca8113 cells. PMID- 17462150 TI - [Expression of drug resistance and metastasis related gene in adenoid cystic carcinoma and mucoepidermoid carcinoma]. AB - OBJECTIVE: To examine the differential gene expression among adenoid cystic carcinoma ACC-M, low metastasis ACC-2 and low differentiated mucoepidermoid carcinoma MEC-1 with gene array, and detect three important genes with real-time quantitative PCR. METHODS: Gene array was used to screen the differential genes, then three important genes were selected and detected by real-time quantitative PCR. RESULTS: The gene contents of MDR-1, MRP-1 and CCD1 were MEC-1 > ACC-M > ACC 2, MEC-1 > ACC-M > ACC-2 and MEC-1 > ACC-M > ACC-2 respectively. CONCLUSIONS: Drug resistance of MEC-1 is related with high content of MDR-1. High metastasis of ACC-M is concerned with high content of MRP-1 and low content of CCD-1. Low metastasis of ACC-2 has a relationship with low content of MRP1. PMID- 17462151 TI - [Clinical trial on intrusion of overerupted maxillary molars with microscrew]. AB - OBJECTIVE: To investigate the imtrusion of overerupted molars with microscrews as anchorage. METHODS: Thirteen adult patients were treated with microscrew anchorage and fixed appliances. Twenty-three overerupted posterior maxillary teeth were intruded. The intrusive movement was investigated on cephalometric radiographs. RESULTS: The molars were intruded and the occlusal plane was corrected successfully in all patients. The treatment period of intrusion was from 5 to 18 months (mean 10.4 months). Significant true intrusion of overerupted maxillary molars, ranged from 0.45 mm to 7.00 mm [mean (2.86 +/- 1.80) mm], was achieved (P < 0.001). The apical root resorption was not clinically significant and the bone level was unchanged. CONCLUSIONS: The microscrew anchorage and fixed appliances were applicable and efficacious for intrusion of overerupted maxillary molars. PMID- 17462152 TI - [Effects of sex hormones on genioglossal muscle activities, estrogen and androgen receptor expression in adult rat]. AB - OBJECTIVE: To explore gene expression of estrogen receptor (ERalpha, ERbeta) and androgen receptor (AR) in genioglossal muscle (GG) of adult male rats, and to investigate the effects of sex hormones on GG activities, ERalpha, ERbeta and AR expression. METHODS: GG samples were collected from 10 healthy adult male rats. Total RNA were extracted and subjected to fluorescent quantitative reverse transcription-polymerase chain reaction (FQ RT-PCR) for quantitative measurement of ERalpha, ERbeta and AR mRNAs. The other 24 male rats were randomly divided into 3 groups: control group, estrogen group (intramuscular injection of estrogen 0.1 mg/kg, twice a week) and androgen group (intramuscular injection of androgen 2.5 mg/kg, twice a week). The electromyographic activities (EMG) and contract tension of GG were investigated after 4-week treatment. The expression of ERalpha, ERbeta and AR was assessed by Western blot. RESULTS: The mRNA expression ratios of AR/GAPDH, ERalpha/GAPDH, ERbeta/GAPDH and ERalpha/ERbeta were (295.80 +/- 127.20), (2042.00 +/- 921.57), (65.96 +/- 29.57) and (36.83 +/- 19.66), respectively. The mRNA level of ERalpha was significantly higher than that of ERbeta (P < 0.01). Compared with the control group, the EMG of GG was intensified in the estrogen group (P < 0.01). GG contractility did not change significantly (P > 0.05), and ERalpha expression in GG was up-regulated by estrogen (P < 0.05); while in the androgen group, the EMG of GG was weakened (P < 0.05). P(t) and P(0) were slightly increased (P > 0.05) and the decline rate of P(0) was markedly quickened (P < 0.05). AR and ERbeta expressions were down-regulated by androgen (P < 0.05). CONCLUSIONS: Both AR and ER were expressed in GG of adult male rats, and ERalpha was expressed more abundantly than ERbeta. Estrogen could greatly improve activities of GG and stimulate the expression of ERalpha. Whereas, androgen could restrain activities of GG, impair its fatigue resistance capacity and inhibit the expression of AR and ERbeta. PMID- 17462153 TI - [Relationship between clinical assessment and patient satisfaction in anterior esthetic restorations]. AB - OBJECTIVE: To analysis the relationship between esthetic characters and satisfaction degree of patients. METHODS: Ninety patients requiring single anterior tooth restoration were treated in clinic with all-ceramic crowns. A visual analogue scale (VAS) was used to assess the satisfaction degree of patients after treatment. Clinical assessments were made by a prosthodontist in characters of feature, surface texture, hue, color distribution, incisal transparency, and individual characterization of all-ceramic crowns. Color difference between the crowns and corresponding natural teeth was tested by Vita Easyshade electrical colorimetric instrument. The relationship between the last two items and patients' satisfaction degree was analyzed. RESULTS: There were significance differences in the different degree among all the six esthetic characters (P < 0.05), except good and moderate degrees in the character of hue (P = 0.553). And color distribution, individual characterization, and surface texture of the crowns showed high correlations with the satisfaction degree of patients (r = 0.485, 0.472, 0.467). Total color-differences, absolute value of hue-differences, and Deltaa between restorations and corresponding natural teeth were related to the satisfaction degree of patients, and Spearman correlation coefficient was -0.434, -0.371, and -0.406 (P < 0.05), respectively. CONCLUSIONS: All the esthetic characters assessed in clinic were related to the satisfaction degrees of patients, especially color distribution, individual characterization and surface texture. Total color-differences and hue-differences were correlated with the satisfaction degrees of patients as well. PMID- 17462154 TI - [Effect of chewing-side preference on activation of parahippocampal gyrus during clenching task]. AB - OBJECTIVE: To explore effects of chewing-side preference (CSP) on cerebral cortical response during clenching task with functional magnetic resonance imaging (fMRI). METHODS: Seven patients with left CSP and 7 patients with right CSP were selected to perform the maximum voluntary clenching in intercuspal position, and block designed blood oxygenation level-dependent (BOLD) fMRI scan covering the whole brain was carried out. RESULTS: There was activation in the parahippocampal gyrus in the four patients among seven with left CSP. But there was no activation in the parahippocampal gyrus among the patients with right CSP. CONCLUSIONS: This research shows that the parahippocampal gyrus, which is associated with human memory and study, is activated during clenching task in the patients with left CSP. PMID- 17462155 TI - [Clinical association of gingipain K-caspase like subdomain expression of Porphyromonas gingivalis with puberty gingivitis]. AB - OBJECTIVE: To detect and compare the activity and intensity of gingipain K (Kgp) caspase like subdomain in culture medium and cell extract of Porphyromonas gingivalis (Pg) isolates in puberty gingivitis and to reveal the possible association of Kgp with puberty gingivitis. METHODS: Thirty-six children of 14 to 17 years old were enrolled in this study. Clinical parameters including gingival index (GI), sulcus bleeding index (SBI) and probing depth (PD) were evaluated. Subgingival plaque samples were collected and Pg isolates were obtained. 16S rRNA PCR was used to confirm Pg clinical isolates. Bacteria were grown in batches of BHI base and harvested at the end of log-phase growth. Culture fractions (culture medium and cell extract) of 10 Pg isolates were performed with SDS-PAGE and Western blot technique using primary antibody against specific Kgp-caspase like subdomain. Activity of Kgp in both samples was detected as well. The data were statistically analyzed using SPSS 11.5 software. The relationship between the Kgp intensity/activity of Kgp and the clinical parameters was statistically analyzed using Spearman correlation coefficient. RESULTS: There was positive correlation between the intensity/activity of Kgp and the clinical parameters (P < 0.05). CONCLUSIONS: The Kgp in clinical isolates of Pg from puberty gingivitis is in complicated forms. Caspase-like molecules with low molecular weight may exist as intracellular functional protein molecules which can affect the interaction between Pg and host. Kgp was contributes in certain degree to the pathogenesis of puberty gingivitis. PMID- 17462156 TI - [Prevalence and related factors of periodontitis in community-dwelling Chinese with diabetes]. AB - OBJECTIVE: To evaluate the periodontal conditions in community-dwelling Chinese with diabetes and analyze some related factors. METHODS: A total of 90 (45- to 84 year-old) patients with diabetes, participating in community-based non communicable diseases management system, were recruited in this study. The examination items on these patients included periodontal measurements performed by pocket probing depth (PPD), attachment loss (AL), sulcus bleeding index (SBI), plaque index (PLI), and blood sugar metabolic level surveillance, as well as structured questionnaire interview. RESULTS: The prevalence of periodontitis was 100%, while 37.1% was diagnosed as having mild periodontitis, 24.4% with moderate, and 38.5% with advanced periodontitis. The AL level in male was significantly higher than that in female. The periodontal destruction was highly correlated with HbA1C value and affected by some socio-behavior factors based on multinomial logistic model. CONCLUSIONS: There is a positive correlation between AL and HbA1C level, and AL and HbA1C should be measured regularly. The patients' periodontal inflammation needs effective control. Priority should be given to the low income population and those with little periodontal knowledge for behavior interventions. PMID- 17462157 TI - [Effect of ozone on the remineralization of enamel in vitro]. AB - OBJECTIVE: To investigate the effect of ozone on remineralization of enamel in vitro. METHODS: Caries-like lesions were prepared in the enamel of extracted cattle incisors with demineralizing solution. The specimens were treated with deionized water, 1000 mg/kg fluoride, 4.5 mg/L ozone, and pH-cycled. The scanning electron microscope (SEM) was used to observe the surface of enamel specimens. The effect of remineralization was assessed by three lesion parameters (fluorescent area, total and average fluorescence), which were measured with confocal laser scanning microscope (CLSM). The data were collected and analyzed by SNK test in ANOVA. RESULTS: SEM observation showed that the surface morphology of the deionized water group and ozone group were similar: porous and atactic; but the surface morphology of fluoride group was smooth and the aperture became smaller. The three parameters in the three groups were as follows, respectively. Delta A were (0.31 +/- 0.28), (1.73 +/- 0.63) and (0.28 +/- 0.19) x 10(4) microm2, Delta TF were (0.42 +/- 0.34), (2.53 +/- 0.73) and (0.48 +/- 0.27) x 10(5), Delta AF were (3.35 +/- 2.55), (46.51 +/- 16.64) and (4.95 +/- 3.05). The fluoride group was significantly greater than deionized water group and ozone group (P < 0.05); but there was no significant difference between the deionized water group and ozone group. CONCLUSIONS: The results of this in vitro study suggest that ozone has no effect on remineralization of enamel blocks. PMID- 17462159 TI - [Establishment and evaluation of a scale method for rating children's behavior in dental clinic in China]. AB - OBJECTIVE: To establish and evaluate a scale method for rating children's behavior in dental clinic in China. METHODS: DV records were taken for 210 children under 8-year-old, who visited the department of pediatric dentistry of Peking University School and Hospital of Stomatology during Jun to Aug in 2004. The Venham's clinic anxiety rating scale and cooperative behavior rating scale were translated into Chinese, and modified according to these records. Twenty four pieces of records were selected randomly, and 11 pediatric dentists and assistants assessed the child behavior in the 24 records. The intraclass correlation coefficient (ICC) and Spearman's correlation coefficient (SCC), which denoted inter-rater reliability and intra-rater reliability, were calculated. RESULTS: The ICC was 0.929 and the SCC was 0.963. CONCLUSIONS: The scale has good inter-rater reliability and excellent intra-rater reliability, and it is competent for rating the children's behavior in dental clinic in China. PMID- 17462160 TI - [In vitro study of the effects of copper ion on osteoclastic resorption in various dental mineralized tissues]. AB - OBJECTIVE: To investigate the effects of copper ion on osteoclastic resorption in various dental mineralized tissues. METHODS: Osteoclasts were separated from long limb bones of neonatal rabbits, and cultured with de-activated human tooth slices and glass slices. The cells in the experiment group were treated with (1 x 10( 14))-1 x (10(-4)) mol/L copper + 10% (volume fraction) fetal calf serum (FCS) + alpha-MEM, while the cells in control group cells were grown in 10% FCS + alpha MEM. Osteoclasts on glass slices were stained by TRAP staining. The absorption pits on tooth slices were observed by inverted phase contrast microscope. The resorbing activity was evaluated with the concentration of calcium in the supernatant liquid of osteoclasts. The ratio between the concentration of calcium in the experiment group and control group was termed as the resorption index. RESULTS: The isolated cells were multinuclear and TRAP positive in cytoplasma. Osteoclasts resorbed teeth slices first on the cementum and dentin. Compared with those on bone slices, the lacunae on the dental slices numbered less, with smaller volume and shallower in depth. Microscopy showed that the number and area of absorption pits formed on treated tissues were less than those on control tissues. The content of calcium in the supernatant liquid decreased in the concentrations of 1 x 10(-14) mol/L - 1 x 10(-4) mol/L copper, especially in the group of 1 x 10(-10) mol/L copper at 3rd day (P < 0.05) and 1 x 10(-4) mol/L, 1 x 10(-10) - 1 x 10(-12) mol/L copper at 7th day (P < 0.05). Their resorption index was lower than one. CONCLUSIONS: Extracellular copper ion can inhibit osteoclastic resorption on dental slices. PMID- 17462161 TI - [Expression of alpha1-adrenoceptor and promoting secretory response by phenylephrine in rabbit submandibular gland]. AB - OBJECTIVE: To investigate the expression and distribution of alpha1-adrenoceptor subtypes in rabbit submandibular gland and the effect of phenylephrine on salivary secretion. METHODS: The expressions of alpha1A-, alpha1B-, and alpha1D adrenoceptor mRNA and protein were investigated by reverse transcription polymerase chain reaction (RT-PCR) and Western blot in rabbit submandibular gland. Immunohistochemical assay was applied to detect the distribution of alpha1A-, alpha1B-, and alpha1D-adrenoceptor and localization of aquaporin 5 in rabbit submandibular gland. Different concentrations of phenylephrine (1 x 10( 8))-(1 x 10(-6)) mol/L were administrated through a polyethylene tube, which was intubated into Wharton's duct of submandibular gland. Heart rate and blood pressure of rabbits were observed during phenylephrine administration. Salivary flow was measured by the length of moist filter paper (35 mm x 5 mm) within 5 min. RESULTS: Alpha1A-, alpha1B-, and alpha1D-adrenoceptor mRNA and protein were expressed in rabbit submandibular gland. Three alpha1-adrenoceptor subtypes were widely distributed in the membrane and cytoplasma of both acinar and ductal cells. Phenylephrine (1 x 10(-7) mol/L, 100 microl) stimulated effectively salivary secretion without inducing significant alteration of blood pressure and heart rate in rabbit. Immunohistochemical assay showed that aquaporin 5 was mainly localized in the apical and lateral plasma membrane in both acinar and ductal cells in unstimulated condition, while the expression of aquaporin 5 was increased after administration of phenylephrine. CONCLUSIONS: Expression of alpha1A-, alpha1B-, and alpha1D-adrenoceptor mRNA and protein was existed in rabbit submandibular gland. Phenylephrine safely and effectively promoted salivary secretion when it was administrated through Wharton's duct of submandibular gland. The mechanism of phenylephrine on salivary secretion may involve in the increase of expression of aquaporin 5 in the apical and lateral plasma membrane in rabbit submandibular gland. This study will hopefully lead to a novel strategy for clinical treatment of dysfunction of submandibular gland. PMID- 17462162 TI - [Effect of aurum coating on corrosion resistance of Ni-Cr alloy]. AB - OBJECTIVE: To evaluate the effect of aurum coating on corrosion resistance of Ni Cr alloy in artificial saliva environment. METHODS: The corrosion potential (E(corr)), self-corrosion current density (I(corr)), and polarization resistance (R(p)) of three alloys were measured using electrochemical methods to compare the difference of corrosion resistance between aurum-coated Ni-Cr alloy and Ni-Cr alloy or Au alloy. Meanwhile, microstructural and phase diffraction was examined with field scanning electromicroscopy (FSEM) and surface chemical analysis was performed by energy diffraction X-ray (EDX). RESULTS: The I(corr) of aurum-coated Ni-Cr alloy was (0.70 +/- 0.20) x 10(-6) A/cm2, which was significantly higher than that of Au alloy (P < 0.05) and lower than that of Ni-Cr alloy (P < 0.05). R(p) of aurum-coated Ni-Cr alloy was (34.77 +/- 12.61) KOmega.cm2, which was higher than that of Ni-Cr alloy (P < 0.05) and lower than that of Au alloy (P < 0.05). The results of FSEM showed that the corrosion resistance of Ni-Cr alloy coated with aurum was better than that of Ni-Cr alloy. The results of EDX indicated that released Ni and Cr of Ni-Cr alloy coated with aurum after test were less than those of Ni-Cr alloy (P < 0.05). CONCLUSIONS: The corrosion resistance of aurum-coated Ni-Cr alloy is higher than that of Ni-Cr alloy. PMID- 17462163 TI - [Clinical application of evidence-based dentistry: Part I. On the need for skills of evidence-based dental practice]. PMID- 17462164 TI - [Rights maintenance and self-discipline of practicing dentist: Part V. Compensate for personal harm in medical treatment dispute]. PMID- 17462165 TI - [A review of etiology and management of sialorrhea]. PMID- 17462166 TI - Additional therapy for young children with spastic cerebral palsy: a randomised controlled trial. AB - OBJECTIVES: To investigate whether, in the short and medium term, additional support by (a) a physiotherapy assistant improved physical function in young children with spastic cerebral palsy and (b) a family support worker improved family functioning. DESIGN: This was a multi-centre randomised controlled trial (RCT) with blinded assessments and a cost-effectiveness analysis. The children studied had spastic cerebral palsy that was the consequence of perinatal adversity. All were less than 4 years old on entry to the study. SETTING: In the child development centre and in the home. PARTICIPANTS: Seventy-six families completed the intervention period. Forty-three families were reassessed 6 months after the end of the intervention and 34 of these after a further 6-month period. INTERVENTIONS: Randomisation was to: (a) a group who received extra physiotherapy from a physiotherapy assistant; (b) a group who received standard physiotherapy; and (c) a group where the child received standard physiotherapy and the family was also visited by a family support worker. Children in all groups continued to receive standard physiotherapy in addition to the study interventions. MAIN OUTCOME MEASURES: The child outcome measures were motor functioning, developmental status and adaptive functioning. The family outcome measures were self-reported maternal stress, level of family needs and parental satisfaction. RESULTS: There was no evidence that additional physical therapy for 1 hour per week for 6 months by a physiotherapy assistant improved any child outcome measure in the short or medium term. Intervention by a family support worker did not have a clinically significant effect on parental stress or family needs. Over the 6 month period the total cost of services for each child ranged from 250 pounds to 6750 pounds, with higher costs associated with children with more severe impairments. No significant relationship was found between measures of intensity of services received by the children and families and the main outcome measures. Low-functioning children, in terms of both motor and cognitive function, were more likely to receive more services in terms of range and frequency. Parents generally reported high satisfaction ratings after all interventions and some stated that the interventions had benefited the child and/or the family. There was therefore a discrepancy between the perceptions of these parents and the objective, quantitative measurements. The family support workers identified a small number of families who were experiencing considerable family problems, but who had not been referred for appropriate support by any other agency. CONCLUSIONS: The findings of this study provide support for the current literature that there was no evidence that additional intervention (in this case by a physiotherapy assistant or family support worker) helped the motor or general development of young children with spastic cerebral palsy. Nor was there any quantitative evidence that providing extra family support helped levels of parental stress and family needs. The implication was that the provision of extra physical therapy does not necessarily improve the motor function of a young child with cerebral palsy and additional family support should not automatically be assumed to be beneficial. In addition, no significant association was found between the intensity of the local services provided and any outcome measure, other than a slight association with lowered family needs. The provision of local services was related to the severity of the child's impairments and not to family difficulties. A small group of families with complex family problems needed more service input. There was a wide range in the costs of services. Research is needed to examine what 'sufficient' levels of provision or therapy might be for which children and which families. A time series of different levels of input and outcomes would provide valuable information for practitioners. It is also recommended that future assessments of therapies of this type adopt a similar multifaceted approach, which is likely to be more suitable than a simple RCT for the evaluation of clinical interventions where the effects are complex. The most appropriate measures of outcome should be used, including assessment of provision of information and emotional support for families. PMID- 17462167 TI - Screening for type 2 diabetes: literature review and economic modelling. AB - OBJECTIVES: To reconsider the aims of screening for undiagnosed diabetes, and whether screening should be for other abnormalities of glucose metabolism such as impaired glucose tolerance (IGT), or the 'metabolic syndrome'. Also to update the previous review for the National Screening Committee (NSC) on screening for diabetes, including reviewing choice of screening test; to consider what measures would be taken if IGT and impaired fasting glucose (IFG) were identified by screening, and in particular to examine evidence on treatment to prevent progression to diabetes in these groups; to examine the cost-effectiveness of screening; and to consider groups at higher risk at which screening might be targeted. DATA SOURCES: Electronic databases were searched up to the end of June 2005. REVIEW METHODS: Literature searches and review concentrated on evidence published since the last review of screening, both reviews and primary studies. The review of economic studies included only those models that covered screening. The new modelling extended an existing diabetes treatment model by developing a screening module. The NSC has a set of criteria, which it applies to new screening proposals. These criteria cover the condition, the screening test or tests, treatment and the screening programme. Screening for diabetes was considered using these criteria. RESULTS: Detection of lesser degrees of glucose intolerance such as IGT is worthwhile, partly because the risk of cardiovascular disease (CVD) can be reduced by treatment aimed at reducing cholesterol level and blood pressure, and partly because some diabetes can be prevented. Several trials have shown that both lifestyle measures and pharmacological treatment can reduce the proportion of people with IGT who would otherwise develop diabetes. Screening could be two-stage, starting with the selection of people at higher risk. The second-stage choice of test for blood glucose remains a problem, as in the last review for NSC. The best test is the oral glucose tolerance test (OGTT), but it is the most expensive, is inconvenient and has weak reproducibility. Fasting plasma glucose would miss people with IGT. Glycated haemoglobin does not require fasting, and may be the best compromise. It may be that more people would be tested and diagnosed if the more convenient test was used, rather than the OGTT. Five economic studies assessed the costs and short-term outcomes of using different screening tests. None examined the long-term impact of different proportions of false negatives. All considered the costs that would be incurred and the numbers identified by different tests, or different cut-offs. Results differed depending on different assumptions. They did not give a clear guide as to which test would be the best in any UK screening programme, but all recognised that the choice of cut-off would be a compromise between sensitivity and specificity; there is no perfect test. The modelling exercise concluded that screening for diabetes appears to be cost-effective for the 40-70-year age band, more so for the older age bands, but even in the 40-49-year age group, the incremental cost-effectiveness ratio for screening versus no screening is only 10,216 pounds per quality-adjusted life-year. Screening is more cost-effective for people in the hypertensive and obese subgroups and the costs of screening are offset in many groups by lower future treatment costs. The cost-effectiveness of screening is determined as much by, if not more than, assumptions about the degree of control of blood glucose and future treatment protocols than by assumptions relating to the screening programme. The very low cost now of statins is also an important factor. Although the prevalence of diabetes increases with age, the relative risk of CVD falls, reducing the benefits of screening. Screening for diabetes meets most of the NSC criteria, but probably fails on three: criterion 12, on optimisation of existing management of the condition; criterion 13, which requires that there should be evidence from high-quality randomised controlled trials (RCTs) showing that a screening programme would reduce mortality or morbidity; and criterion 18, that there should be adequate staffing and facilities for all aspects of the programme. It is uncertain whether criterion 19, that all other options, including prevention, should have been considered, is met. The issue here is whether all methods of improving lifestyles in order to reduce obesity and increase exercise have been sufficiently tried. The rise in overweight and obesity suggests that health promotion interventions have not so far been effective. CONCLUSIONS: The case for screening for undiagnosed diabetes is probably somewhat stronger than it was at the last review, because of the greater options for reduction of CVD, principally through the use of statins, and because of the rising prevalence of obesity and hence type 2 diabetes. However, there is also a good case for screening for IGT, with the aim of preventing some future diabetes and reducing CVD. Further research is needed into the duration of undiagnosed diabetes, and whether the rise in blood glucose levels is linear throughout or whether there may be a slower initial phase followed by an acceleration around the time of clinical diagnosis. This has implications for the interval after which screening would be repeated. Further research is also needed into the natural history of IGT, and in particular what determines progression to diabetes. An RCT of the type required by NSC criterion 13 is under way but will not report for about 7 years. PMID- 17462168 TI - The effectiveness and cost-effectiveness of cinacalcet for secondary hyperparathyroidism in end-stage renal disease patients on dialysis: a systematic review and economic evaluation. AB - OBJECTIVES: To establish the effectiveness and cost-effectiveness of cinacalcet for the treatment of secondary hyperparathyroidism (SHPT) for people on dialysis due to end-stage renal disease (ESRD). DATA SOURCES: Electronic databases were searched up to February 2006. REVIEW METHODS: Included randomised controlled trials (RCTs) on the clinical effectiveness of cinacalcet for SHPT in ESRD were critically appraised, had relevant data extracted and were summarised narratively. A Markov (state transition) model was developed that compared cinacalcet in addition to current standard treatment with phosphate binders and vitamin D to standard treatment alone. A simulated cohort of 1000 people aged 55 with SHPT was modelled until the whole cohort was dead. Incremental costs and quality-adjusted life-years (QALYs) were calculated. Extensive one-way sensitivity analysis was undertaken as well as probabilistic sensitivity analysis. RESULTS: Seven trials comparing cinacalcet plus standard treatment with placebo plus standard treatment were included in the systematic review. A total of 846 people were randomised to receive cinacalcet. Cinacalcet was more effective at meeting parathyroid hormone (PTH) target levels (40% vs 5% in placebo, p < 0.001). In those patients meeting PTH targets, 90% also experienced a reduction in calcium-phosphate product levels, compared with 1% in placebo. Significantly fewer people treated with cinacalcet were hospitalised for cardiovascular events, although no difference was seen in all-cause hospitalisation or mortality. Significantly fewer fractures and parathyroidectomies were also seen with cinacalcet. Findings on all patient-based clinical outcomes were based on small numbers. The authors' economic model estimated that, compared to standard treatment alone, cinacalcet in addition to standard care costs an additional 21,167 pounds and confers 0.34 QALYs (or 18 quality-adjusted weeks) per person. The incremental cost-effectiveness ratio (ICER) was 61,890 pounds/QALY. In most cases, even extreme adjustments to individual parameters did not result in an ICER below a willingness-to-pay threshold of 30,000 pounds/QALY with probabilistic analysis showing only 0.5% of simulations to be cost-effective at this threshold. Altering the assumptions in the model through using different data sources for the inputs produced a range of ICERs from 39,000 pounds to 92,000 pounds/QALY. CONCLUSIONS: Cinacalcet in addition to standard care is more effective than placebo plus standard care at reducing PTH levels without compromising calcium levels. However, there is limited information about the impact of this reduction on patient-relevant clinical outcomes. Given the short follow-up in the trials, it is unclear how data should be extrapolated to the long term. Together with the high drug cost, this leads to cinacalcet being unlikely to be considered cost-effective. Recommendations for future research include obtaining accurate estimates of the multivariate relationship between biochemical disruption in SHPT and long-term clinical outcomes. PMID- 17462169 TI - The clinical effectiveness and cost-effectiveness of gemcitabine for metastatic breast cancer: a systematic review and economic evaluation. AB - OBJECTIVES: To assess the clinical effectiveness and cost-effectiveness of gemcitabine, used in combination with paclitaxel, as a second-line treatment for people with metastatic breast cancer who have relapsed following treatment with anthracycline-based chemotherapy. DATA SOURCES: Electronic databases were searched from inception to March 2006. Clinical advisers were also consulted. REVIEW METHODS: A systematic review of the literature was undertaken to appraise the clinical and cost-effectiveness of gemcitabine. A Markov state transition model was developed for the economic evaluation. RESULTS: The systematic review identified only one randomised controlled trials (RCT), and this has not yet been fully published. The methodological quality and quality of reporting of the included trial were assessed to be poor using standard criteria, but this may be due to the lack of information in the limited publications rather than being a fair reflection of the trial's quality. This RCT compared gemcitabine and paclitaxel therapy with paclitaxel monotherapy in 529 patients with metastatic breast cancer who had previously received anthracyclines, but no prior chemotherapy for metastatic breast cancer. Approximately 71% of the gemcitabine/paclitaxel patients survived for 1 year, compared with 61% of the paclitaxel group. The hazard ratio showed a 26% lower chance of survival in the paclitaxel group, and time to progressive disease was also shorter in this group. The overall response rate was higher in the gemcitabine/paclitaxel group than in the paclitaxel group. Adverse events, particularly neutropenia, were more common with gemcitabine/paclitaxel combination therapy than with paclitaxel therapy alone. The economic model was run for a simulation of 1000 patients, assuming that chemotherapy continued until patients' disease progressed. This base-case analysis found an incremental cost-effectiveness ratio (ICER) of 58,876 pounds per quality-adjusted life-year (QALY) gained and 30,117 pounds per life-year gained. The model was re-run with treatment restricted to a maximum of six cycles per patient, reflecting normal practice. This yielded an ICER of 38,699 pounds per QALY gained and 20,021 pounds per life-year gained. CONCLUSIONS: The review of clinical effectiveness is based on data from a single RCT that has not yet been fully published. While only tentative conclusions can be drawn from this, the evidence may indicate that treatment with gemcitabine and paclitaxel confers an improved outcome for patients in terms of survival and disease progression, but at the cost of increased toxicity. An economic model developed for this review reflects high costs per QALY for this treatment combination. The base-case analysis shows high ICERs, with costs per QALY gained close to 60,000 pounds. Adopting a more realistic treatment protocol, with chemotherapy limited to a maximum of six cycles, gives a more favourable cost-effectiveness estimate. However, this was still higher than would usually be considered to be a cost effective treatment from the NHS's perspective. Future research recommendations include an update of this review in 12-18 months' time, by which time the included RCT should be fully published. It would also be useful to compare gemcitabine with currently used treatments for metastatic breast cancer, including capecitabine and vinorelbine. PMID- 17462170 TI - A systematic review of duplex ultrasound, magnetic resonance angiography and computed tomography angiography for the diagnosis and assessment of symptomatic, lower limb peripheral arterial disease. AB - OBJECTIVES: To determine the diagnostic accuracy and cost-effectiveness of duplex ultrasound (DUS), magnetic resonance angiography (MRA), and computed tomography angiography (CTA), as alternatives to contrast angiography (CA), for the assessment of lower limb peripheral arterial disease (PAD). DATA SOURCES: Ten electronic databases were searched in April 2004, with an update in May 2005. Six key journals and bibliographies of included studies were also searched and experts in the field were consulted. REVIEW METHODS: Data extraction and quality assessment were performed in duplicate. Data were analysed according to test type and diagnostic threshold. For the economic analysis, a decision tree was developed and a probabilistic sensitivity analysis performed to incorporate statistical uncertainty into the cost-effectiveness analysis. RESULTS: A total of 113 studies met the inclusion criteria (including six economic evaluations). For the detection of stenosis greater than 50% in the whole leg, contrast-enhanced (CE) MRA (14 studies) had the highest diagnostic accuracy, with sensitivity ranging from 92 to 99.5% and specificity from 64 to 99%. Two-dimensional (2D) time-of-flight (TOF) MRA (11 studies) was less accurate, with sensitivity ranging from 79 to 94% and specificity from 74 to 92%. 2D phase-contrast (PC) MRA (one study) had a sensitivity of 98% and specificity of 74%. CTA (seven studies) also appeared slightly inferior to CE MRA, with a sensitivity ranging from 89 to 99% and specificity from 83 to 97%, but better than DUS (28 studies), which had a sensitivity ranging from 80 to 98% and specificity from 89 to 99%. There was some indication that CE MRA and DUS were more accurate for detecting stenoses/occlusions above the knee than below the knee or in the pedal artery. The four studies of patient attitudes strongly suggested that patients preferred CE MRA to CA. CA was considered the most uncomfortable test, followed by CE MRA, with CTA being the least uncomfortable. Half of the patients (from a sample who did not suffer from claustrophobia and had no metallic implants) expressed no preference between undergoing TOF MRA or DUS; most of those who did express a preference favoured TOF MRA. In the 55 studies identified for adverse events, MRA was associated with the highest reported proportion. However, the most severe adverse events were more common in patients undergoing CA; although these were rare for both tests. The economic evaluation showed DUS dominated the other alternatives when the whole leg was assessed, by presenting higher effectiveness at a lower cost per quality-adjusted life-year (QALY; i.e. 13,646 pounds per QALY). When the assessment was limited to a section of the leg, either above the knee or below the knee, 2D TOF MRA was the most cost-effective preoperative diagnostic strategy. The incremental cost per QALY for below-the-knee comparisons was equal to 37,024 pounds when 2D TOF MRA was compared with DUS. For above-the knee comparisons, 2D TOF MRA presented the lowest cost and slightly lower effectiveness compared with CE MRA, with a cost per QALY equal to 13,442 pounds. CONCLUSIONS: The results of the review suggest that CE MRA has a better overall diagnostic accuracy than CTA or DUS, and that CE MRA is generally preferred by patients over CA. Where available, CE MRA may be a viable alternative to CA. The only controlled trial suggested that the results of DUS were comparable to those of CA, in terms of surgical planning and outcome. This finding conflicts with the results of diagnostic accuracy studies, which reported poor estimates of accuracy for DUS in comparison with CA. There was insufficient evidence to evaluate the usefulness of CTA for the assessment of PAD, particularly newer techniques. The results of the economic modelling suggest that for PAD patients for whom the whole leg is evaluated by a preoperative diagnostic test, DUS dominates the other alternatives by presenting higher effectiveness at a lower cost per QALY. However, when the analysis of stenosis is limited to a section of the leg, either above the knee or below the knee, 2D TOF MRA appears to be the most cost effective preoperative diagnostic strategy. Further research is needed into a number of areas including the relative clinical effectiveness of the available imaging tests, in terms of surgical planning and postoperative outcome. PMID- 17462171 TI - Shared care model for mental health: is it a viable option for pakistan ? PMID- 17462172 TI - Superficial chemical peeling with salicylic Acid in facial dermatoses. AB - OBJECTIVE: To determine the effectiveness of salicylic acid chemical peeling in common dermatological conditions affecting face in people with predominant Fitzpatrick skin type IV and V. DESIGN: An interventional, quasi-experimental study. PLACE AND DURATION OF STUDY: Military Hospital, Rawalpindi and PAF Hospital, Sargodha, from July 2002 to June 2003. PATIENTS AND METHODS: A total of 167 patients of either gender, aged between 13 to 60 years, having some facial dermatoses (melasma, acne vulgaris, postinflammatory hyperpigmentations, freckles, fine lines and wrinkles, post-inflammatory scars, actinic keratoses, and plane facial warts) were included. A series of eight weekly hospital based peeling sessions was conducted in all patients under standardized conditions with 30% salicylic acid. Clinical improvement in different disorders was evaluated by change in MASI score, decrease in the size of affected area and % reduction in lesions count. McNemar test was applied for data analysis. RESULTS: Majority of the patients showed moderate to excellent response. There was 35% to 63% improvement (p<0.05) in all dermatoses. Significant side effects, as feared in Asian skins were not observed. CONCLUSION: Chemical peeling with salicylic acid is an effective and safe treatment modality in many superficial facial dermatoses. PMID- 17462173 TI - Thyroid dysfunction in infertile women. AB - OBJECTIVE: To determine the frequency of thyroid dysfunction in infertile women referred for thyroid evaluation. DESIGN: A retrospective case-control study. PLACE AND DURATION OF STUDY: This study was carried out at Centre for Nuclear Medicine (CENUM), Mayo Hospital, Lahore, from July 2003 to December 2006. PATIENTS AND METHODS: Age matched infertile (n=140 each) and fertile women (n=152 each) referred to CENUM for thyroid evaluation were investigated for incidence of hyperthyroidism (TSH < 0.03 mIU/L), hypothyroidism (TSH < 0.03 mIU/L) and thyroid autoimmunity (antithyroid peroxidase antibody titer>20 IU/L). Serum free T4 (FT4), free T3 (FT3) and antithyroid peroxidase antibody (TPO-Ab) was determined by radioimmunoassay (RIA) and TSH by immunoradiometric assay (IRMA). RESULTS: Most of the infertile women (89.3%), like control women (93.4%), were euthyroid. The difference of overall thyroid dysfunction was not statistically significant in infertile and control women (10.7% vs. 7.9%; p=0.395). The same was true for incidence of hyperthyroidism (4.3% vs. 5.3%; p=0.701) as well as hypothyroidism (6.4% vs. 2.6%; p=0.104). In infertile women, the incidence of hypothyroidism (6.4%) was slightly higher as compared to hyperthyroidism (4.3%). In euthyroid women of both groups, mean FT4, FT3 and TSH levels were significantly higher (p < 0.05) in infertile women and double number of them had serum TSH>2.5 mIU/L compared to fertile women (31.2% vs. 15.6%; p<0.01). Similarly, more infertile women were TPO-Ab positive (titer>20 IU/L) than control women (7.2% vs. 1.4%; p < 0.05). CONCLUSION: Increased incidence of high normal TSH and raised TPO-Ab titer indicate relatively more frequent occurrence of compensated thyroid function in infertile women than normal women of reproductive age. This necessitates considering them a subgroup of women in which all aspects of pituitary-thyroid axis should be thoroughly investigated than merely TSH testing. PMID- 17462174 TI - HELICOBACTER PYLORI, HEPATITIS VIRUSES A, C, E, ANTIBODIES AND HBsAg - PREVALENCE AND ASSOCIATED RISK FACTORS IN PEDIATRIC COMMUNITIES OF KARACHI. AB - OBJECTIVE: To document the prevalence of Helicobacter pylori (H. pylori), Hepatitis A virus (HAV), Hepatitis C virus (HCV), Hepatitis E virus (HEV) antibodies and Hepatitis B virus surface antigen (HBsAg), in the pediatric age group of low socioeconomic urban communities of Karachi and to identify risk factors associated with these infections. DESIGN: Cross-sectional survey. PLACE AND DURATION OF STUDY: Three selected squatter settlements of Karachi during April 2002 to December 2004. PATIENTS AND METHODS: Three hundred and eighty children, ages 5 months to 15 years were investigated. Venous blood samples were collected and questionnaire filled on sociodemographic characteristics (family income, number of dependents in the family, area of living, number of people per room per house, and number of children sharing bed with parents and siblings). Gastrointestinal symptoms were recorded. Anti-HAV IgG (Hepatitis A virus IgG antibody), anti-HCV (Hepatitis C virus antibody), anti-HEV (Hepatitis E antibodies) and HBsAg, were analyzed by enzyme immunoassays (EIAs). Samples were also screened for anti-HIV1/2 (human immunodeficiency virus 1 and 2 antibodies by EIA. IgG antibodies against H. pylori were detected by immunochromatography. RESULTS: A correlation between increasing age and seroconversion was seen for hepatotropic viruses. At 14 years and above,100% of the children were found to be positive for anti-HAV, 26% for anti-HEV, and 1.4%, for anti-HCV while HBsAg was positive in 1.9%. H. pylori infection did not show a significant increase with age. Both anti-HAV and anti-H. pylori were present simultaneously in 30% of the population investigated. CONCLUSION: With age, increasing number of children acquired antibodies against hepatotropic viruses and H. pylori. Occurrence of HBsAg and anti-HEV at a later age suggests horizontal, rather than vertical transmission. PMID- 17462175 TI - Endometriosis: frequency and correlation between symptomatology and disease stage. AB - OBJECTIVE: To determine the frequency of endometriosis in general gynecological and infertile women presenting to hospital and to correlate symptomatology and stage of disease. DESIGN: Cross-sectional study. PLACE AND DURATION OF STUDY: Gynecology Department of Mother and Child Health Center, Pakistan Institute of Medical Sciences, Islamabad, from March 2000 to March 2002. PATIENTS AND METHODS: Fifty patients with diagnosis of endometriosis on laparoscopy were included in the study. The degree of pain symptoms was graded according to 1-4 point verbal rating scale designed by Biberoglu and Behrman. Endometriosis was staged according to Revised American Fertility Society (R-AFS) scoring on laparoscopy. Correlation coefficient-Spearman rank order correlation test was applied to analyze data. RESULTS: The frequency of endometriosis in infertile women was 24% (33) and in women with general gynecological complaints, it was 23% (17). There was a significant positive correlation between chronic pelvic pain and R-AFS scoring. Increasing severity of pelvic pain was also positively correlated with presence of endometrioma and complete obliteration of pouch of Douglas. No correlation between dysmenorrhoea and R-AFS scoring could be detected. Dysmenorrhoea was strongly and positively correlated with the presence of superficial implants. Dyspareunia was found to have positive correlation with R AFS score and also with complete obliteration of pouch of Douglas. CONCLUSION: The frequency of endometriosis in infertile women was 24% (33) and in women with general gynecological complaints, it was 23% (17). Chronic pelvic pain and dyspareunia had strong positive correlation with R-AFS score while dysmenorrhoea had no such correlation. PMID- 17462176 TI - Demographic and clinical profile of patients with complicated unsafe abortion. AB - OBJECTIVE: To describe the demographic and clinical profile of patients admitted as a result of complicated unsafe abortion. DESIGN: A case series. PLACE AND DURATION OF STUDY: The study was carried out in the Department of Obstetrics and Gynaecology, Jinnah Hospital, Lahore from August 2001 to July 2002. PATIENTS AND METHODS: Patients admitted with complicated unsafe abortion were evaluated regarding age, parity, marital and educational status, indication for abortion, method used, qualification of abortion providers, contraceptive usage, complications and death rate in abortion seekers. Descriptive statistics was used for describing variables. RESULTS: Fiftynine patients were admitted with complicated unsafe abortion. The mean age was 29 years, 95% were married and multiparous, 40% had secondary and higher education, 85% approached unqualified abortion providers who used instrumentation in more than 40% of cases for termination of pregnancy resulting in visceral trauma. More than 50% were using contraception and 5% died due to postabortion complications. CONCLUSION: Unsafe abortion is a major health problem. The associated morbidity is much higher than mortality. This study focus on the need of postabortion care and easy accessibility to contraception to improve quality of health. PMID- 17462177 TI - Justifying the clinical use of fresh frozen plasma - an audit. AB - OBJECTIVE: To determine the appropriateness of fresh frozen plasma (FFP), uses in various haematological and clinical disorders, with reference to the British Committee for Standards in Haematology (BCSH) guidelines through an audit. DESIGN: Descriptive study. PLACE AND DURATION OF STUDY: Department of Haematology and Transfusion Medicine, Shalamar Hospital, Lahore, from June 2001 to June 2004. SUBJECTS AND METHODS: The data was collected from June 2001 to June 2004 from the request forms ordered by the clinicians for the transfusion of FFP at the Department of Haematology and Transfusion Medicine, Shlamar Hospital, Lahore. A total of 2075 healthy blood donors donated their whole blood for the preparation of fresh frozen plasma (FFP). All blood donors were screened for anti HCV, HBsAg, VDRL and HIV. Those 2075 FFP units were prepared on high-speed centrifuge and were rapidly stored at -30oC freezer. A total of 587 patients were transfused 2075 units of FFP for various clinical disorders. The percentage of FFP units, transfused appropriately and inappropriately, as defined by BCSH guidelines, was estimated. RESULTS: Out of 2075 FFP units, 335 (24.41%) FFP units were transfused to patients suffering from bleeding due to disseminated intravascular coagulation (DIC), 306 (22.30%) units used for massive transfusion and surgical bleeding, 236 (17.20%) units for bleeding due to chronic liver disease, 202 (14.72%) units used to control bleeding due to coagulation factor deficiencies, 84(6.12%) units for thromobotic thrombocytopenic purpura (TTP), 75(5.46%) units prior to liver biopsy to correct prolonged prothrombin time (PT), 72(5.24%) units for haemorrhage due to haemolytic disease of newborn (HDN) and 62(4.51%) units to control bleeding due to warfarin overdosage, 425(60.45%) units used for nutritional support and hypovolaemia replacement, 131(18.63%) units for the reversal of prolonged INR in the absence of bleeding due to warfarin, 92 (13.08%) units used in ICU to correct prolonged PT without bleeding due to Vitamin K deficiency and 55(7.82%) units for chronic liver disease (CLD) to correct prolonged PT and APTT in the absence of bleeding. In summary, 1372 (66.12%) FFP units were appropriately and 703 (33. 88 %) were inappropriately used. CONCLUSION: In conclusion, 33.88% FFP was inappropriately used mainly due to lack of awareness of international guidelines and ignorance of risks. PMID- 17462178 TI - Clinical and neurological study of women with precatamenial epilepsy. AB - OBJECTIVE: To describe clinical features and seizure dispersion in precatamenial/pericatamenial epilepsy patients. DESIGN: Case series. PLACE AND DURATION OF STUDY: Department of Neurology (formerly Neuropsychiatry), Jinnah Postgraduate Medical Centre (JPMC), Karachi, from July 1991 to November 2001. PATIENTS AND METHODS: Present study included 33 untreated pericatamenial (n: 23, age: 12-40 years, menstrual cycles: 147) and precatamenial (n: 10, age: 13-32 years, menstrual cycles: 70) epileptics with tonic-clonic seizures. Clinical features and seizure dispersion were evaluated during premenstruation, menstruation, and postmenstruation phases. RESULTS: Women with precatamenial epilepsy had highly significant mean phase day seizures during premenstruation versus other phases, whereas women with pericatamenial epilepsy did not show any significant variations. Premenstrual seizures were found significantly more and others as significantly less in % number in precatamenials compared to those in pericatamenials. Furthermore, precatamenial epileptics with primary generalized seizures were significantly higher in % number and secondary generalized seizures as significantly lower against those in pericatamenial epileptics. All precatamenial epileptics under study had incontinence compared to 65% pericatamenial epileptics that had incontinence. Majority of the patients in both groups showed post-ictal headache. CONCLUSION: The present report describes the extent of exacerbation of premenstrual tonic-clonic seizures with clinical features. These investigations may help in understanding partly the complexity of catamenial/precatamenial/ pericatamenial/noncatamenial seizures, and similarities and dissimilarities between pericatamenial and precise precatamenial seizures. PMID- 17462179 TI - Threshold parameters for choroidal vessel closure with the photosensitizer, lambda 27. AB - OBJECTIVE: To determine the threshold photodynamic therapy parameters, required for choroidal vessel closure, with the photosensitizer, Lambda 27, in Dutch belted rabbits, using fluorescein angiography and histopathology. DESIGN: A pre clinical experiment. PLACE AND DURATION OF STUDY: Department of Ophthalmology at the Tulane University Health Sciences Center, between June 2001 - July 2002. PATIENTS AND METHODS: Dutch belted rabbits were divided into two groups. The first group was injected intravenously with an aqueous solution of Lambda 27 at 1mg/kg. Saline was injected intravenously into the second group (controls). Approximately 5 minutes after injection of Lambda 27, a diode laser, mounted on Zeiss 30 SL-M slit lamp, emitting light at a wavelength of 719 nm was used to apply photodynamic therapy to the fundus. Lesions were placed on the choroid of the rabbits at a fixed spot size of 1.5 mm, using powers ranging from 50 to 120 mW, for a duration of 5 to 80 seconds. Choroidal vessel closure was documented by fluorescein angiography, the following day, and by light microscopy, after the animals were sacrificed. RESULTS: Choroidal vessel closure was documented using fluorescein angiography for all of the lesions placed at fluence of 22.7 J/cm2 and above. Histopathology also confirmed this finding. No choroidal vessel closure was seen in the rabbits treated with fluences less than 22.7 J/cm2 or in the control group. CONCLUSION: This experiment establishes threshold parameters for choroidal vessel closure using the photosensitizer, Lambda 27. PMID- 17462180 TI - Anaphylaxis during anesthesia - role of skin prick testing in diagnosing multiple sensitivities. AB - The true incidence of anaphylactic reactions and their associated morbidity and mortality remain poorly defined. This is due to uncertainties in reporting accuracy and exhaustivity. A 23 years old male developed severe anaphylaxis under general anesthesia with cardiovascular collapse, bronchospasm and angio-edema. He was promptly managed with intravenous Adrenaline, Hydrocortisone, colloid solutions and Promethazine. The temporal relation suggested Atracurium to be the cause. However, skin prick test, performed 8 weeks later, showed a very strong (+++) positive test for Atracurium and mild sensitivity (+) with Suxamethonium. No sensitivity to Nalbuphine, Thiopentone or Pancuronium was noted. The relevant information was endorsed in patient's medical record sheet for his future exposure. PMID- 17462181 TI - Unusual metastases of hepatocellular carcinoma (hcc) to bone and soft tissues of lower limb. AB - Hepatocellular carcinoma metastasizing to bones and soft tissues of extremities is an unusual occurrence. The present report describes two such cases, where this unusual happening was the presenting feature. Role of immunohistochemistry for diagnosis is emphasized. PMID- 17462182 TI - Hepatic hydatid cyst presenting as anaphylaxis. AB - A young soldier was brought to the emergency of Combined Military Hospital (CMH), Multan in a collapsed state with two day's history of chest pain. He was resuscitated and placed on ventilator. While the rest of examinations were normal, ultrasound examination of the abdomen revealed a cystic mass in liver most likely to be hydatid. The patient was fully conscious the next day and was put on tab albendazole (200 mg BID). CT scan of the abdomen revealed a large cystic mass having inner undulating wall with watery content diagnosed as hydatid cyst in right lobe of liver. Since anaphylaxis was considered consequent to hydatid cyst perforation, surgery was carried out. Approximately 100 ml of haemorrhagic fluid was aspirated and 10% hypertonic saline instilled. After re aspiration, cyst cavity was opened and endocyst completely removed. Portion of ectocyst projecting away from the liver edge was also excised. He made an uneventful postoperative recovery and was discharged with the advice to continue tab albendazole 200mg BID for four weeks and weekly follow-up in surgical OPD. He is doing well now. PMID- 17462183 TI - Isolated malignant peripheral nerve sheath tumor of retroperitoneum. AB - A case of an isolated Malignant Peripheral Nerve Sheath Tumor (MPNST) of the retroperitoneum without neurofibromatosis is presented. The tumor was located deep in the retroperitoneum with metastasis to the ribs. It was surgically removed. Patient was further managed with radiotherapy and chemotherapy. MPNST in such a location is very unusual. PMID- 17462184 TI - Acute small bowel obstruction secondary to ileal endometrioma. AB - Pelvic endometriosis, a common disease of the female genital tract, may also affect the bowel, especially the rectosigmoid colon. Involvement of small intestine occurs very infrequently. A case of small bowel obstruction caused by ileal endometriosis is reported. PMID- 17462185 TI - Compound palmar ganglion with carpal tunnel syndrome. AB - Compound palmer ganglion is an uncommon condition characterized by a swelling in the distal part of volar aspect of wrist and communicating with another swelling over palm across the flexor retinaculum. It commonly results from tuberculous tenosynovitis. It may lead to carpal tunnel syndrome and need surgical excision with division of flexor retinaculum. PMID- 17462186 TI - Delayed obstructive jaundice secondary to bullet in common hepatic duct. AB - A young male presented with obstructive jaundice and recurrent attacks of acute cholangitis. He had sustained gunshot wound in abdomen 9 years back. Laparotomy was done at that time and the bullet was found left within right lobe of liver, as it was inaccessible and possible benefit to take it out was considered less than leaving it in-situ. The patient recovered and did well for 7 years. After 7 years of latency, he developed recurrent attacks of acute cholangitis. The reason of this delayed presentation was probably the migration of the bullet from its original place to the biliary tree i.e. common hepatic duct. PMID- 17462187 TI - Intestinal perforation due to an ingested foreign body. AB - It is a case description of intestinal perforation with foreign body that was arrested in hernia- trapped intestinal loop and causing perforation, which was explored and removed under local anesthesia. Literature on gut perforation due to foreign body ingestion is reviewed. PMID- 17462188 TI - Neurological manifestations as presenting feature in dengue Fever. AB - Dengue fever (DF), caused by dengue viruses, presents classic dengue fever, dengue hemorrhagic fever, and dengue shock syndrome. A young man presented with 5 days history of fever, body aches, headache, vomiting and one day history of drowsiness and a generalized tonic clonic fit. He was afebrile and hypotensive with platelets count of 24 x 109/l, and serum IgM DENV positive. Computerized tomography scan of the brain revealed intracerebral haemorrhages in both cerebral hemispheres. He was managed with supportive care and 20% mannitol infusion, his headache and drowsiness rapidly improved. Platelet count rose to 60 x 109/l on 2nd day and 199 x 109/l on 7th day of admission. He remained afebrile during hospital stay and was discharged from hospital on 8th day of admission, when he was almost asymptomatic. PMID- 17462189 TI - Esophago-pleural fistula with pulmonary tuberculosis. AB - A patient with clinical diagnosis of pulmonary tuberculosis and right sided hydropneumothorax found to have fistulous connection of pleura with oesophagus as evidenced by upper GI endoscopy and intrapleural instillation of radio-opaque dye. He was managed with intercostal chest tube drainage, antibiotics, antitubercular regimen and nasogastric tube feeding, resulting in eventless recovery subsequently. PMID- 17462190 TI - Hepatitis and the healthcare worker - a pakistani perspective. AB - Hepatitis B (HBV) and C (HCV) virus infections are the most important causes of chronic liver disease, and the biggest health challenges facing the developing world today. Pakistan is in the intermediate HBV and HCV prevalence area. The health care worker (HCW) is at the forefront of this battle to control this epidemic. From physicians, nurses, to the para-medical staff, the HCW constantly places himself in potential danger, by attending to infected patients. This article reviews the literature available so far on the potential risk of transmission of HBV and HCV both to and from the HCW and makes recommendations for the prevention of such transmission in our working environment. PMID- 17462191 TI - Glanzmann'S thrombasthenia: presentation and management of a rare bleeding disorder. PMID- 17462192 TI - [Prevalence of renal artery stenosis in patients with critical limb ischemia]. AB - BACKGROUND AND OBJECTIVE: Our purpose was to evaluate the prevalence of renal artery stenosis in patients with critical limb ischemia and to study any clinical or laboratory indicator that could predict this association. PATIENTS AND METHOD: One hundred consecutive patients with critical limb ischemia evaluated by angiogram were included in the study from January to July 2003. Cardiovascular risk factors and renal function were analyzed. RESULTS: One hundred angiographic studies have been analyzed. Thirty nine (39%) of our patients had some type of pathology of the renal artery but the rest, 61 (61%), had normal and healthy renal arteries. In 5 patients, a bilateral renal pathology was found. Severe disease (> 60% stenosis, bilateral or renal occlusion) was present in 15 cases including 6 occlusions. Once we compared the patients with healthy renal arteries with the patients with different degrees of stenosis, we did not appreciate significant differences in hypertension, diabetes, coronary disease or smoking habit, nor with laboratory data such as creatinine, urea, c-reactive protein, total cholesterol or atherogenic index. No differences were found either comparing patients with normal renal artery with patients with bilateral pathology or with unilateral occlusion. CONCLUSIONS: There is a high prevalence of renal artery pathology in patients with critical limb ischemia although we have not found any clinical or laboratory factors useful to identify them. PMID- 17462193 TI - [Tobacco smoking and sputum smear conversion in pulmonary tuberculosis]. AB - BACKGROUND AND OBJECTIVE: To analyze if cigarette smoking delays the sputum smear conversion in pulmonary tuberculosis. PATIENTS AND METHOD: Ninety eight patients were diagnosed with pulmonary tuberculosis. Patients were all not immunosuppressed, infected by human immunodeficiecy virus (HIV) or drug resistant. Sixty four of them were smokers with a pack-year index (standard deviation) of 33.69 (23.12). Delayed sputum smear conversion (DC) was considered when 2 positive sputum culture results were obtained in the second month of anti tuberculous treatment and was associated with the following variables in 2 groups: a) total group (in which all the patients were included): age, sex, smoking habits, risk factors (alcohol consumption, diabetes mellitus, immunosuppression, drug addicion, malnutrition), time with symptoms, radiologic presentation and bacterial load, and b) smokers: age, sex, risk factors, time with symptoms, radiologic presentation, bacterial load and pack-year index. For the statistical analysis, chi2 test, Student t test and logistic regression model were used, considering the dependant variable DC. RESULTS: In the total group, 17 patients (17.3%) had DC, 16 of them had a history of smoking and in the univariate analysis it was associated with: alcohol consumption, time with symptoms, radiologic presentation as bilateral cavitary infiltrates and smoking habits. The logistic regression analysis showed an association with smoking habits (odds ratio = 9.8; p = 0.03) and bilateral cavitary infiltrates (odds ratio = 3.61; p = 0.02). In the group of smokers, DC was associated in the univariate analysis with the female sex. CONCLUSIONS: Smoking habits delay sputum conversion in patients with pulmonary tuberculosis not associated with HIV and non-resistant bacilli. According to these results it is necessary to assist smoking cessation in patients who are receiving antituberculous treatment. PMID- 17462194 TI - [Feasibility of the sentinel lymph node technique in cervical and vulvar cancers]. AB - BACKGROUND AND OBJECTIVE: Our goal was to investigate the feasibility of identifying the sentinel lymph node (SLN) in patients undergoing surgery for cervical and vulvar carcinomas. We also evaluated the advantages that such procedure can offer in this kind of tumors. PATIENTS AND METHOD: We detected the sentinel lymph node through preoperative lymphoscintigraphy (Technetium-99m colloid) and intraoperative visualization with blue dye by a hand-held or laparoscopic gamma-probe in 7 patients with invasive cervical carcinoma (clinical stage Ib1) and in 6 patients with vulvar carcinoma (clinical stages Ib and II) . RESULTS: At least one sentinel lymph node was identified in each patient. SNL was more commonly found in patients with invasive cervical carcinoma. We observed neoplastic infiltration in 3 nodes, all of them corresponding to cases of vulvar carcinoma. CONCLUSIONS: Sentinel lymph node identification is a feasible technique in the management of vulvar and cervical carcinomas. Our preliminary data show that SLN in gynecological carcinomas have similar uses as in other anatomical sites, identifying women in whom lymph node dissection can be avoided. PMID- 17462195 TI - [Treatment of ischemic nephropathy]. PMID- 17462196 TI - [Multicentric study of research on confidentiality]. PMID- 17462197 TI - [Current knowledge on pathogeny, diagnosis and treatment of hepatitis B]. PMID- 17462198 TI - [Adverse effects of selective serotonin reuptake inhibitors use during the third trimester of pregnancy and prevention guidelines]. AB - Selective Serotonin Reuptake Inhibitors (SSRIs) have become the drug of choice for the treatment of depression and have shown to be effective in the treatment for other mental disorders. Recently, several articles have reported about the adverse effects observed in newborns after maternal exposure to these drugs during the last trimester of pregnancy. In this work, a review of literature is presented, regarding the above mentioned adverse effects. Moreover, some guidelines for the rational use of these drugs during the last trimester of pregnancy and for the management of prenatally exposed newborns are provided. PMID- 17462199 TI - [Eosinophilic esophagitis]. AB - Eosinophilic esophagitis is characterised for a dense infiltration of the esophagus by eosinophilic leukocytes. The disease's origin is a local reaction to different antigens of which the patient presents previous sensitization, acquired by digestive, inhaled or even epicutaneous exposure. The esophagus contains different cellular types resident in its structure, with capability to participate in the capture, processing and antigens' presentation to T lymphocytes, which could initiate a T helper 2-type immunological response mostly mediated by interleukin-5, with a possible T helper 1-type component. Local production of immunoglobulin E could also participate in the pathophysiology of eosinophilic esophagitis, and for this reason, this disease can be considered a mixed-humoural and cell-mediated immunological disturbance. Studies directed to identificate responsible allergens must consider test for determine immunoglobulin E-mediated reactions as well as cell-mediated hyper-responsiveness responses. Main symptom of eosinophilic esophagitis are dysphagia and esophageal food impactations, which are conditioned by endoscopic alterations and motor disturbances objectively demonstrated by manometric recorders. Eosinophil and mast cell's activation and degranulation against responsible antigens cause damage over esophageal epithelium and dynamic disturbances over neuromuscular components in esophageal wall. Therapies proposed for eosinophilic esophagitis include control of antigen exposition, endoscopic dilation of stenosis and drugs with antieosinophilic effect; in this group topical steroids can be outlined for the capacity of them to restore the histology and the esophageal motility in parallel to vanishment of inflammation. PMID- 17462200 TI - [Asymptomatic intrapelvic cyst associated to total hip arthroplasty]. PMID- 17462201 TI - [Reversible posterior leukoencephalopathy in a patient with systemic lupus erythematosus]. PMID- 17462202 TI - [Ethmoid-sphenoidal sinusitis]. PMID- 17462204 TI - CT scan and appendicitis: inadequate population. PMID- 17462206 TI - Management of penetrating neck injuries: zone II. PMID- 17462207 TI - Development and pilot testing of an OSCE for difficult conversations in surgical intensive care. AB - OBJECTIVES: To describe the development and results of an Objective Structured Clinical Exam (OSCE) for leading family conferences in the surgical intensive care unit (SICU). DESIGN: Pilot demonstration and reliability assessment. SETTING: General surgery residency program at a major academic teaching hospital. PARTICIPANTS: PGY-2 and PGY-4 categorical general surgery residents (n=8). RESULTS: The SICU Family Conference OSCE consists of two 20-minute stations, one requiring residents to lead an end-of-life discussion and the other to disclose an iatrogenic complication. Actual case scenarios and trained actors were used; the examinations were videotaped in a standardized setting. Two professional raters as well as the participating actors assessed each resident performance using rating tools developed for each station and based on guiding principles gleaned from the literature. Resident debriefings and evaluation surveys were also conducted. Resident perception of the OSCE overall was positive. Analysis of the videotapes revealed the need for greater standardization of the actors' roles. The rating tools showed strong internal consistency (0.77-0.85), but inter rater agreement of scores was generally low (<0.70) within rater groups. Family actors consistently gave residents higher global assessment scores than did the professional raters. Second- and fourth-year residents scored equally well on the examination. CONCLUSIONS: This pilot provided residents with a positive learning experience and valid formative feedback. Case materials developed for each station served their function well. More work in actor and rater training is needed before the examination scores can be reliably used in summative evaluation. PMID- 17462208 TI - The effect of preoperative clopidogrel on bleeding after coronary artery bypass surgery. AB - INTRODUCTION: Clopidogrel treatment is associated with a reduction in thrombotic complications in coronary stent placement, improved outcome after acute coronary syndromes, and decreased mortality in patients with coronary artery disease. The purpose of this study was to analyze the effect of preoperative clopidogrel exposure on bleeding complications, blood transfusions requirements, and reoperations in patients undergoing coronary artery bypass grafting (CABG). PATIENTS AND METHODS: This study included 320 patients from a single institution that underwent an isolated CABG who were discharged between July 2003 and June 2004. The cohort of 320 patients was classified into 3 groups. The control group consisted of 255 patients that did not receive clopidogrel or stopped clopidogrel 7 days before surgery but were treated with aspirin instead. Clopidogrel I consisted of 25 patients that were taking clopidogrel within 3 days of surgery, and Clopidogrel II consisted of 40 patients that were taking clopidogrel 4 to 7 days before surgery. Patients were compared based on preoperative data (age, gender, use of clopidogrel, preoperative hemoglobin, and ejection fraction), intraoperative data (cross-clamp time), postoperative data (chest tube output, rate of reoperation, units of transfused blood, length of stay in the intensive care unit, and length of intubation). RESULT: There were no significant differences among the 3 groups concerning age, sex, ejection fraction, or preoperative hemoglobin. There were no differences in length of intensive care unit stay and length of intubation among the 3 groups of patients. Patients in the clopidogrel I group had more units of blood transfused than either the control or the Clopidogrel II group (p=0.027). There is also a trend toward more chest tube output in clopidogrel I group compared with the control group. Fifteen patients (4.6%) of the total group required reoperation secondary to bleeding: 2 (8.0%) in the Clopidogrel I group, 2 (5%) in the clopidogrel II group, and 11 (4.3%) in the control group (p=0.41). CONCLUSION: This study demonstrated that clopidogrel within 3 days preoperatively increases the requirement for blood transfusion in patients undergoing CABG. Waiting more than 3 days after the last dose of clopidogrel decreases blood transfusion requirements. There is also a trend toward more postoperative bleeding for those patients that took clopidogrel within 3 days before their CABG. The reoperation rate of patients that took clopidogrel within 3 days of their procedure required almost twice as many reoperations as the patients that did not take clopidogrel. PMID- 17462209 TI - Understanding statistical tests. PMID- 17462210 TI - Giant colonic diverticulum: an unusual abdominal lump. AB - Giant colonic diverticulum is a rare complication of diverticular disease of the colon and is thought to result, in most cases, from a "ball-valve" effect. The presentation and clinical course can be variable and confusing. The most common symptoms are abdominal pain and a palpable abdominal lump, with many patients presenting acutely with complications such as perforation and peritonitis. Preoperative diagnosis requires a high degree of suspicion and needs to be differentiated from sigmoid volvulus, caecal volvulus, intestinal duplication cyst, pneumatosis cystoidis intestinalis, and similar conditions. A plain x-ray and computed tomography (CT) scan of the abdomen shows a huge air-filled cyst termed "balloon sign" and confirms the diagnosis. The barium enema shows a communication with the bowel in most cases. In view of the high incidence of complications, treatment is advised even in asymptomatic cases and consists of excision of the cyst with resection of the adjacent colon with primary anastomosis. This treatment would, in most cases, be a sigmoid colectomy. Percutaneous drainage and Hartmann's procedure may be appropriate in some cases who present with a well-formed abscess or gross fecal peritonitis, respectively. A case is described, and the literature is reviewed. PMID- 17462211 TI - Minimally invasive parathyroidectomy complicated by pneumothoraces: a report of 4 cases. AB - The traditional approach to primary hyperparathyroidism has been a bilateral neck exploration for evaluation of all four parathyroid glands. With the advent of minimally invasive surgery, minimally invasive parathyroidectomy has become a popular approach for the treatment of parathyroid adenomas. Though exceedingly rare, pneumothorax formation is a potential complication following this procedure. In this paper, we report four cases of pneumothorax following minimally invasive parathyroidectomies. The commonality in all these cases was positioning with extreme neck hyperextension. Additional risks in three patients were dissection in the superior mediastinum, traction on the thyrothymic ligament, and a low-lying inferior parathyroid gland. One patient developed a pneumothorax prior to dissection along the superior mediastinum. This suggests that further risk factors may be heat conduction from the electrocautery and total intravenous anesthesia with spontaneously breathing of the patient. PMID- 17462212 TI - Use of pre-operative Tc99m-Sestamibi scintigraphy and intraoperative parathyroid hormone monitoring to eliminate neck exploration in mediastinal parathyroid adenocarcinoma. AB - A 66-year-old white woman was found to have an elevated serum calcium and parathyroid hormone (PTH) on routine health evaluation. Physical examination was unremarkable as was ultrasonography of the neck. A sestamibi parathyroid scan revealed abnormal uptake in the anterior mediastinum. Computed tomography of the chest demonstrated an anterior mediastinal mass compatible with a parathyroid adenoma but no neck masses. The patient underwent mediastinoscopy that was converted to a median sternotomy to fully access the mass. The mass was completely resected with surrounding thymus gland. Frozen section confirmed that excised tissue was parathyroid gland in origin. An intraoperative PTH obtained 20 minutes after specimen removal showed a decrease of more than 50% from preoperative levels. The strategy for initial surgery for hyperparathyroidism when a sestamibi scan is "positive" in the mediastinum (only) is a point of some controversy. Traditional recommendations have been to "clear the neck" of abnormal parathyroid tissue before undertaking a more morbid sternotomy. Mediastinoscopy was attempted to remove the mediastinal lesion and to avoid a sternotomy. Preoperative Tc99m sestamibi scintigraphy, frozen section histology, and intraoperative PTH monitoring permitted the authors to conclude that neck exploration was unnecessary. PMID- 17462214 TI - Recent evidences of the use of 18F-fluorodeoxyglucose positron emission tomography in the management of colorectal cancer. AB - Positron emission tomography is considered a potentially useful diagnostic tool in the management of a variety of malignancies. It has been used for staging, evaluation of recurrent and metastatic disease, evaluation of tumoral response to neoadjuvant therapy, and providing prognostic information. However, some data are available about its definitive role in the colorectal cancer population. In this review, the most recent evidences and future perspectives are presented for the use of the 18F-fluorodeoxyglucose positron emission tomography in the management of patients with colorectal cancer. PMID- 17462215 TI - Enrique Finochietto: the legacy of surgery in Argentina. PMID- 17462216 TI - Improving your new resident orientation program. A perspective from the University of Missouri-Columbia. PMID- 17462217 TI - Personal and surgical experiences during a rotation in San Cristobal, Dominican Republic. PMID- 17462218 TI - The teaching of surgery in the undergraduate curriculum--reforms and results. AB - In the past decade, the teaching of surgery in the undergraduate curriculum has undergone considerable changes in quantity, mode and method of delivery. This is a result of the radical reforms of higher education, the health service and the undergraduate medical curriculum. These reforms are often interrelated and are occurring in conjunction with major changes in healthcare delivery. In this article we discuss this reorganisation, the rationale behind it and the impact on surgical teaching. PMID- 17462219 TI - Psychological considerations in face transplantation. PMID- 17462221 TI - Ethical considerations in face transplantation. PMID- 17462223 TI - Sociological considerations in face transplantation. PMID- 17462225 TI - An audit and comparative analysis of the kidney transplantation programme in Burma. AB - BACKGROUND: The first renal transplant operation to be carried out by a Burmese transplant team was undertaken at the Yangon General Hospital, Burma on the 8th of May 1997. Since then 21 further kidney transplantations have been performed in hospitals in Yangon (Rangoon) and Mandalay. Burma was renamed as Myanmar in 1989. The Myanmar national renal transplant programme was introduced and performed as a special project organised by the Ministry of Health. Unfortunately little is known in other countries about the programme's development or the outcomes of the patients treated. OBJECTIVES: In August 2003, the author visited Myanmar with the intention of evaluating the progress that this programme had made in the last six years. MATERIAL AND METHODS: Retrospective data concerning all recipients and donors involved in the Myanmar renal transplant programme was obtained from the patient databases and through personal meetings with staff at the Urology and Nephrology units in Yangon and Mandalay. RESULTS: All 22 cases were blood related living donor transplants. There were 14 males and 8 female donors with a mean age of 36.5 (28-53) years and 13 male and 9 female recipients with a mean age of 38.3 (19-60) years included in the programme. There was a 100% survival rate of all 22 donors and a 95% (n = 21) recipient survival rate recorded in the programme up to September 2003. The one-year graft survival rate is 95% (n = 21) and the overall graft survival rate until September 2003 is 91% (n = 20), the mean follow-up was 2 years and 4 months. CONCLUSION: The success of this programme is extremely impressive considering the turbulent socio-economic climate in this developing country. The next steps for Myanmar are to organise a domestic registry system for transplants, the organisation of personnel to take responsibility for data collection, distribution and greater communication with other transplant units. PMID- 17462226 TI - CRP in acute appendicitis--is it a necessary investigation? AB - BACKGROUND: Appendectomy is one of the commonest procedures in surgery. In spite of various investigations used to improve the accuracy of diagnosis, the rate of normal appendices removed is still about 15-30%. Many studies have investigated the role of C-reactive protein (CRP) in acute appendicitis, but with conflicting results. METHODS: In a prospective, double blind study, blood for the measurement of serum C-reactive protein was collected pre-operatively from 192 children before going to the operating theatre for appendectomy. The histopathology was grouped into positive (acute appendicitis) and negative (normal appendix) and this was correlated with CRP values. RESULTS: CRP was normal in 14 out of 33 negative explorations (normal appendix on histopathology). The specificity and sensitivity of serum CRP was 42% and 91% respectively. The predictive value of a positive (raised CRP) and negative (normal CRP) test is 88% and 48% respectively. CONCLUSION: We conclude that neither raised nor normal CRP value is helpful in the diagnosis of acute appendicitis. CRP is not a good tool for helping the surgeon make the diagnosis of appendicitis and it should not be measured in suspected appendicitis. PMID- 17462227 TI - Preoperative anthropometric analysis of the cleft child's face: a comparison between groups. AB - This study examines the facial surfaces of different groups of cleft babies aged up to eleven months, prior to any corrective surgery, with the aim of identifying and assessing differences in their facial morphology. Measurements of standard anthropometric landmarks were made on plaster casts taken pre-operatively of the faces of babies presenting for surgical correction of lip and palate deformities. Periorbital and mid face measurements show no difference between control and cleft groups. The nasal base is shown to be wider in infants with cleft lip on both cleft and non-cleft sides. Upper lip measurements indicate shortening, lateral deviation and superolateral rotation. A diagrammatic model is proposed to illustrate facial anatomy in control and cleft individuals. Modifications to the technique are proposed. Further study in this field is suggested, in order to produce a comprehensive database of facial morphological changes in the cleft and non-cleft populations both pre-operatively and over time, with a view to producing a potential redefinition of the cleft syndrome based on anthropometric principles. PMID- 17462228 TI - Life after definitive treatment for children with Hirschsprung's disease. AB - PURPOSE: The aim of this study was to investigate the long-term outcomes after definitive surgical correction for children with Hirschsprung's Disease (HD) and the psychosocial impact HD has on the child and family. METHODS: A clinical-based database of seventy-two children and young people aged between one to twenty-four years with HD, along with their families were investigated. This study involved the development of a condition-specific questionnaire in order to assess the functional and psychosocial outcomes for children with HD at different age groups, combined with parental perception of their child's condition in the long term. RESULTS: The greatest functional problem after definitive surgery for HD was faecal soiling (n = 29/38: 76.3%). Children < or = 12 years experienced more embarrassment, distress/discomfort and family difficulties (n = 53/72; 73.6%) due to bowel dysfunctioning (such as faecal soiling) in comparison to children 12 years (n = 19/72; 26.3%) (p < 0.05). Young adults with HD (> 12 years) remained confident and 62.5% hopeful about their future with HD. Nine (12.5%) of the parents reported that HD had a negative impact on their marital relationship due to the daily stressors. Yet, 58.3% (n = 42) families remain confident and 70.8% (n = 42) hopeful about their child's future with HD. CONCLUSION: Bowel functioning and psychosocial distress improves with increasing age and parental and medical professional support. Psychosocial difficulties found in the child and family with HD are condition-specific--thus improving complications such as faecal soiling will further enhance better psychosocial adjustment. PMID- 17462229 TI - Why do orthopaedic surgeons ignore the medial patellofemoral ligament? AB - The medial patellofemoral ligament (MPFL) is a condensation of the medial capsule of the knee joint. In the past two decades dissection studies have shown that it extends from the superomedial border of the patella to the femoral epicondyle, at or immediately above the adductor tubercle. MRI and operative studies have revealed that it is almost invariably damaged by lateral patellofemoral dislocation. Current surgical management of such dislocations may involve imbricating the torn medial capsule and parapatellar retinaculum back onto the medial border of the patella. If the medial patellofemoral ligament is torn at or near the femoral attachment, as the latest MRI and operative studies demonstrate it frequently is, then this medial reefing procedure will not be successful in restoring normal anatomy and function. Here we review the anatomy and function of the MPFL, its role in patellar dislocation and as well as surgical treatment for patellar dislocation. PMID- 17462230 TI - Does laparoscopic surgery offer adequate clearance in rectal cancer?--A discussion. AB - Currently in the UK, the national institute of clinical excellence (NICE), only advocates laparoscopic surgery for rectal cancer as part of commissioned clinical trials. Laparoscopic teaching, training and techniques have evolved greatly and offer many benefits to patients, whilst remaining technically demanding to surgeons still on the slope of the learning curve. Can such minimally invasive techniques be used with the same results as open surgery in the treatment of rectal cancer? Are laparoscopic colorectal surgeons able to achieve the same clearance of tumours and so avoid recurrence at the same rate compared to conventional techniques? The discussion to follow, aims to shed some light on such questions and briefly review some of the literature. If laparoscopic anterior resections and abdominoperineal resections achieve the same results as open procedures, then should these techniques be more widely taught and practised? Surely the peri-operative cost of these laparoscopic procedures does not over shadow the potential outcome from much less traumatic surgery? PMID- 17462231 TI - Robotics and surgery: a long-term relationship? AB - This article introduces robotic surgical systems by explaining the shortcomings of traditional laparoscopic surgery, and how these new systems have been developed to address them. This is followed by a descriptive section of robotic systems past and present and their use in different surgical specialities. Finally, we discuss advances that are planned for the development of current systems and the future role of robotics in surgery. PMID- 17462232 TI - A review of the epidemiology, pathogenesis and management of tetanus. AB - Tetanus remains an important disease worldwide. In the United Kingdom, the elderly and intravenous drug users are at particular risk of acquiring clinical tetanus. Tetanus is associated with a high morbidity and mortality. Once the diagnosis of tetanus is suspected, intensive management is necessary. In this article we review the history, epidemiology, microbiology, clinical features, mode of transmission, pathogenesis, differential diagnosis, management, complications and prevention of this life threatening disease. PMID- 17462234 TI - Patient selection for facial transplantation I: Anatomical and surgical considerations. PMID- 17462236 TI - Patient selection for facial transplantation II: Psychological considerations. PMID- 17462238 TI - Patient selection for facial transplantation III: Ethical considerations. PMID- 17462240 TI - A fatal case of MRSA septicaemia and gallbladder empyema. AB - Methicillin-resistant Staphylococcus aureus (MRSA) has become a major problem worldwide and is associated with significant morbidity and mortality. It is a particular issue in surgical patients especially vascular surgery involving prosthetic implants. We present an unusual case of MRSA gallbladder empyema and septicaemia in a previous vascular surgical patient. Teranishi et al reported a case of acute cholecystitis caused by MRSA in the Japanese literature but as far as we are aware this is the first case reported in the English literature. PMID- 17462241 TI - Intussusception in a young adult with bloody diarrhoea. AB - Bloody diarrhoea is a common presenting symptom in acute surgery, medicine and also in out-patient clinics. Intussusception, although a common cause of small bowel obstruction and rectal bleeding in infants, is rare in adults. In contrast to paediatric cases, where the majority can be treated by pneumatic or hydrostatic reduction, the vast majority of adult cases of intussusception are due to a tumour of some kind and will require operation and resection. Diagnostically plain radiology, CT, ultrasound and endoscopy are useful since the clinical picture is often unclear. We present a case of a young man who presented with acute bloody diarrhoea. The diagnosis was made colonoscopically and he was treated surgically. The case underlines the need for surgeon to maintain an open mind when investigating unexceptional symptoms. PMID- 17462242 TI - An appendiceal faecolith in a 24-year-old man. PMID- 17462243 TI - An unusual surgical presentation of Munchausen's syndrome. PMID- 17462244 TI - The increasing significance of how to learn motor skills. AB - The training of surgeons has traditionally been an apprenticeship. In recent times it has become increasingly difficult for trainees to acquire operative skills. The acquisition and refinement of these technical skills by the surgical trainee are central to surgical teaching. Much work has been done looking into the theories of motor skill learning, working on the assumptions that if they are understood then tests may be developed to be used as skill predictors. Two schools of thought exist as to the use of neuropsychological factors in learning fine motor skills but all agree that practice and instructional feedback are essential for the correct acquisition of these essential surgical skills. With the current climate of shortened training and new guidelines its even more important to understand skill acquisition and assist in the learning of new surgical skills. PMID- 17462245 TI - Improving hospital communication: do mobile phone networks hold the key? PMID- 17462246 TI - Is UK surgical training in crisis? A trainee's perspective. PMID- 17462252 TI - The search for safer surgery. PMID- 17462253 TI - Surgical training at the crossroads. PMID- 17462254 TI - Animal experiments and the development of surgical therapies. PMID- 17462255 TI - Life after the tsunami: perspectives from a UK trained, Sri Lankan born surgical trainee. PMID- 17462256 TI - Ertapenem versus ceftriaxone and metronidazole as treatment for complicated intra abdominal infections. AB - BACKGROUND: Prompt surgical intervention supplemented by appropriate antimicrobial therapy is usually required for successful treatment of complicated intra-abdominal infections. The objective of this study was to further evaluate the efficacy and safety of ertapenem relative to ceftriaxone/metronidazole as treatment for complicated intra-abdominal infections. METHODS: Adult patients with intra-abdominal infections requiring surgery were eligible for this open label randomized trial comparing ertapenem 1 g daily with ceftriaxone 2 g daily plus metronidazole 30 mg/kg/day. The primary efficacy outcome was the clinical response rate in clinically and microbiologically evaluable participants at the test-of-cure (TOC) visit 2 weeks after discontinuation of therapy. All treated patients were included in the safety analysis. RESULTS: Participant demographics, disease characteristics, and duration of therapy in both treatment groups were generally similar. Escherichia coli was the most commonly isolated baseline pathogen, recovered in 52% of cases in each treatment group. Favorable clinical responses were achieved at TOC in 143 (96.6%) of 148 ertapenem recipients and in 146 (96.7%) of 151 ceftriaxone/metronidazole recipients. The frequencies of drug related adverse events, most commonly nausea, diarrhea, vomiting, and elevated platelet count, were generally comparable in both treatment arms. Four ertapenem recipients (1.8%) and one ceftriaxone/metronidazole recipient (0.4%) experienced serious drug-related adverse events. CONCLUSIONS: In this study, ertapenem and ceftriaxone/metronidazole were comparably effective treatments for adult patients with complicated intra-abdominal infections. PMID- 17462257 TI - Tigecycline is efficacious in the treatment of complicated intra-abdominal infections. AB - BACKGROUND: Empiric treatment of complicated intra-abdominal infections (cIAI) represents a clinical challenge because of the diverse bacteriology and the emergence of bacterial resistance. The efficacy and safety of tigecycline (TGC), a first-in-class, expanded broad-spectrum glycylcycline antibiotic, were compared with imipenem/cilastatin (IMI/CIS) in patients with cIAI. METHODS: In this prospective, double-blind, phase 3, multinational trial, patients were randomly assigned to intravenous (i.v.) TGC (100 mg initial dose, then 50 mg every 12 h) or i.v. IMI/CIS (500/500 mg every 6 h) for 5-14 days. Clinical response was assessed at the test-of-cure (TOC) visit (14-35 days after therapy) for microbiologically evaluable (ME) and microbiologically modified intent-to-treat (m-mITT) populations (co-primary efficacy endpoint populations in which cure/failure response rates were determined). RESULTS: Of 817 mITT patients (i.e., received > or = 1 dose of study drug), 641 (78%) comprised the m-mITT cohort (322 TGC, 319 IMI/CIS) and 523 (64%) were ME (266 TGC, 256 IMI/CIS). Patients were predominantly white (88%) and male (59%) with a mean age of 49 years. The primary diagnoses for the mITT group were complicated appendicitis (41%), cholecystitis (22%), and intra-abdominal abscess (11%). For the ME population, clinical cure rates at TOC were 91.3% (242/265) for TGC versus 89.9% (232/258) for IMI/CIS (95% CI -4.0, 6.8; P<0.001). Corresponding clinical cure rates within the m-mITT population were 86.6% (279/322) for TGC versus 84.6% (270/319) for IMI/CIS (95% CI -3.7, 7.5; P<0.001 for noninferiority TGC versus IMI/CIS). The most commonly reported adverse events for TGC and IMI/CIS were nausea (17.6% TGC versus 13.3% IMI/CIS; P=0.100) and vomiting (12.6% TGC versus 9.2% IMI/CIS; P=0.144). CONCLUSIONS: TGC is efficacious in the treatment of patients with cIAIs and TGC met per the protocol-specified statistical criteria for noninferiority to the comparator, IMI/CIS. PMID- 17462258 TI - A comparison of clinical judgment vs the modified Alvarado score in acute appendicitis. AB - AIMS: To investigate the value of the modified Alvarado score (MAS) in helping Accident & Emergency (A&E) doctors decide which patients with suspected acute appendicitis need surgical referral. METHODS: 11,258 patients presented to a University Hospital A&E Department over a two-month period; 82 were triaged as 'abdominal pain' or 'suspected appendicitis'. Ten patients self-discharged prior to seeing a doctor. The remaining case notes (72) were reviewed and MAS's calculated. The Alvarado guidelines suggested an MAS > or = 5 (high) needed admission and an MAS < 5 (low) excluded appendicitis and was appropriate for discharge. RESULTS: Two patients had proven pancreatitis and were excluded. 24/70 patients were admitted for suspected appendicitis; all were referred by the A&E doctor (sensitivity 100%) but only 12 had a high MAS (sensitivity 50%). Twelve patients were therefore admitted despite having a low MAS on retrospective analysis. 46/70 patients were discharged (none re-presented with the same complaint) of which 40/46 were sent home without surgical referral (specificity 87%), but only 44/46 patients discharged had a low MAS (specificity 96%). CONCLUSIONS: It is more important to refer every case that needs referral (sensitivity) than to discharge those not needing referral (specificity). We cannot exclude the possibility that morbidity would result were the MAS used in lieu of clinical judgment in deciding whether referral is necessary in cases of suspected acute appendicitis. PMID- 17462259 TI - Emerging trends in microsurgical breast reconstruction: deep inferior epigastric artery perforator (DIEP) and the superior gluteal artery perforator (SGAP) flaps. AB - Breast reconstruction is an important adjunct in the treatment of breast cancer. Many reconstructive options exist, however autologous tissue remains the gold standard. One drawback to autologous reconstruction methods is the potential for flap donor site morbidity. Recent advances in microsurgical techniques include the development of perforator flaps, including the Deep Inferior Epigastric Artery Perforator flap (DIEP) and the Superior Gluteal Artery Perforator (SGAP) flaps. Harvest of these flaps attempt to minimize the impact on the donor site and thereby reduce the incidence of donor site complications. This article will review of the indications, advantages and drawbacks to the use of perforator flaps in breast reconstruction surgery. PMID- 17462260 TI - Angiogenesis and the tumour hypoxia response in prostate cancer: a review. AB - The formation of new blood vessels, angiogenesis, is a highly-regulated active process that is critical for the development of the normal and malignant prostate. The vascular endothelial growth factor (VEGF) system assumes a critical role in the angiogenic process. Angiogenesis is a prerequisite for the expansion of solid tumours beyond 1-3 mm3 and is stimulated in response to a hypoxic environment. This review discusses the process of angiogenesis and the key angiogenic mediator, VEGF, and their role in tumour progression and metastasis. A better understanding of the mechanisms behind angiogenesis will ultimately lead to the development of new anti-angiogenic agents in the management of prostate cancer. PMID- 17462261 TI - The use and abuse of antibiotics in elective colorectal surgery: the saga continues.... AB - BACKGROUND: The role of antibiotic prophylaxis in preventing post-operative complications in patients undergoing elective colorectal surgery is well established. Despite evidence that a single-dose prophylaxis is sufficient, the duration of antibiotic use in clinical practice is highly variable and surveys have identified persistent patterns of antibiotic abuse in elective colorectal surgery. MATERIALS AND METHODS: We conducted a retrospective review of all patients who underwent elective colorectal surgery between 1998 and 2002 at the American University of Beirut Medical Center. A survey among general surgeons in Lebanon was also performed to investigate the pattern of antibiotic prophylaxis used in such cases. The MEDLINE database (1966-2004) was searched for English language articles and abstracts on antimicrobial use in elective colorectal surgery. Papers cited in relevant primary articles were also reviewed. Data were extracted and reviewed by all authors. RESULTS: Two hundred and eleven matching patient-records were identified. A triple regimen including metronidazole, ampicillin and an aminoglycoside was the most commonly used preoperative prophylactic method. Patients received post-operative antibiotics for a mean of 6.66+/-2.62 days. The mean duration of post-operative antibiotic prophylaxis used by the interviewed surgeons was 4.31+/-1.08 days. CONCLUSIONS: Our study confirms that even when strong evidence exists, surgeons fail to adhere to antibiotic prophylaxis guidelines. This pattern is not unique to Lebanon but is shared to a large extent by surgeons around the world. Adherence to published guidelines and improved education of surgeons are essential to the delivery of cost-effective medical practice. PMID- 17462262 TI - Errors in surgery. AB - Making errors is part of normal human behaviour. However when errors have significant consequences or occur in high risk industries they become of paramount importance. There has been little research in why and how errors occur in the healthcare industry. Errors occur throughout healthcare, but in particular, surgery as a high risk speciality. Surgery is a dynamic speciality with a milieu of possible mishaps waiting to happen. So to understand and prevent errors in surgery we must explore this intricate multi-cogwheel process. This article will summarise the epidemiology of surgical errors, factors which influence them in the patient pathway, explain concepts and models of why errors occur, technical skill error assessment and possible strategies to prevent or reduce surgical errors. Practicing surgery in the new millennium will embrace new innovations, medications, technologies, equipment, operations, all which aim to improve the treatment and care of patients. However we must remember with this constant evolution in healthcare the error goalposts are forever moving, so we must be vigilant not to take our eye off the error ball. PMID- 17462263 TI - The need for technical skills assessment in surgery. PMID- 17462264 TI - The teaching of surgery in the undergraduate curriculum--reforms and results. AB - In the past decade, the teaching of surgery in the undergraduate curriculum has undergone considerable changes in quantity, mode and method of delivery. This is a result of the radical reforms of higher education, the health service and the undergraduate medical curriculum. These reforms are often interrelated and are occurring in conjunction with major changes in healthcare delivery. In this article we discuss this reorganisation, the rationale behind it and the impact on surgical teaching. PMID- 17462265 TI - Practical sculptural training for the plastic surgeon. AB - For the past 25 years I have been working as a portrait sculptor and educator, developing a scientific approach and protocol to train the eye to see accurately and fully understand the complexity of 3-D form. The methods developed over this period, aim to unravel the visual complexity of the human head and to close the gap between the forms that are seen and that which the hand can re-create. The method has been put to test in teaching programmes with surgeons, artists and non artists. PMID- 17462266 TI - Trainees advisory group for the international campaign to revitalise academic medicine: have your say in shaping the future of medical education, research and clinical practice. PMID- 17462268 TI - The editor's diary: 7/7-21/7. PMID- 17462269 TI - Screening for disease--the good--the bad and the thoughtful. PMID- 17462270 TI - Use of vein graft as a tendon sheath substitute following tendon repair: an innovative technique in tendon surgery. AB - OBJECTIVES: This is a new technique for managing tendon repair that may improve the results of existing methods. METHODS: In a prospective randomized clinical trial, 210 patients were divided into two groups of test and control. All patients had flexor tendon injuries, involving zone 2. They were new or old tendon injuries or complications of previous repairs. In the test group (105 patients), a modified Kessler repairing of tendons with 3-0 prolene was used, followed by a core suture of running 6-0 nylon or prolene epitendinous suture. After the tendon repair, a segment of vein through which the tendon had been passed before or a patch of vein, as a tendon sheath substitute, was used to repair the sheath defects. The results in a span of six months of follow-up were compared with those of the control group whereon 105 patients were operated under the conventional technique--the modified Kessler method. RESULTS: We assessed the results by measuring the range of motion of the MCP joint in the follow-up period and we graded them as excellent, good, fair and poor. In the test group we had 86% excellent, 11% good, 3% fair and 0% poor results, and in the control group, 0% excellent, 12% good, 38% fair and 50% poor results. The differences were significant (p<0.005). CONCLUSIONS: Our preliminary results appeared encouraging when compared with the outcomes achieved by the conventional tendon repair technique. As this technique reduces the adhesion formation, improves tendon nourishment, and decreases the need of intensive physiotherapy, it may substitute the conventional one and become a standard technique in the future. PMID- 17462271 TI - Terminal events in women dying of advanced breast cancer: improving the care for terminal breast cancer patients in Africa. AB - We studied the terminal events preceding death in all patients dying in hospital over a period of 10 years. Hepatomegaly, massive ascites, cachexia, jaundice, massive pleural effusion and haemoptysis were the most frequent terminal events. The predominance of visceral-related terminal events is unexplained but may be related to site-specific metastasis and premorbid organ pathology. PMID- 17462272 TI - Improving outcomes in urological cancers: the impact of "multidisciplinary team meetings". AB - INTRODUCTION: In order to improve the outcomes of urological cancers, guidelines published by the National Institute of Clinical Excellence encourage the management of cancer patients by specific Multi-Disciplinary Teams (MDTs) with discussion of cancer patients at MDT Meetings. The aim of this prospective study was to examine the changes in management resulting from review at MDT Meetings in our unit. METHODS: Over a six month period 124 cancer cases were discussed at 10 meetings. Prior to the meetings consultants completed a form stating their proposed management and whether they thought this would be changed after discussion. At the meeting histological, radiological and clinical data were reviewed and a collective decision about the optimal treatment was made. Any changes were recorded. RESULTS: Two of 124 cases had their clinical management changed as a result of the meeting. These were identified (amongst 10 others) as potential 'change cases' prior to the meeting. Four changes were made to histological reports and 1 to radiology; none of these affected clinical management. CONCLUSION: Discussion of cancer cases at MDMs made no difference to the clinical management in over 98% of cases. Consultants correctly identified cases requiring discussion, indicating that a selective rather than blanket approach would be appropriate. This has the potential to reduce the considerable costs involved without affecting patient care. PMID- 17462273 TI - Primary extrahepatic abdominal hydatidosis. AB - INTRODUCTION: Most of the abdominal hydatid cysts occur in liver. Extrahepatic hydatid cyst is usually secondary to rupture (operative and non-operative) of the hepatic hydatid cyst. Primary extrahepatic hydatid cysts are rare and only a few sporadic cases have been reported. MATERIAL AND METHODS: One hundred and eighty three patients with abdominal hydatid cysts managed surgically from January 1998 to December 2003 were evaluated retrospectively. Twelve (6.5%) patients had only extrahepatic abdominal involvement. RESULTS: The cysts were present in spleen (2.2%), pancreas (1.1%), peritoneum and pelvis (1.6%), gallbladder (0.6%), mesocolon (0.6%) and adrenal (0.6%). DISCUSSION AND CONCLUSIONS: It is difficult to diagnose extrahepatic echinococcosis as it usually is not suspected. Symptoms are related to size, location or ensuing complication of the cyst. It should be strongly suspected in differential diagnosis of all abdominal cysts especially in an endemic area. PMID- 17462274 TI - The systemic inflammatory response syndrome and cardiopulmonary bypass. AB - Cardiac surgery using cardiopulmonary bypass (CPB) provokes a systemic inflammatory response. This is mainly triggered by contact activation of blood by artificial surfaces of the extracorporeal circuit. Although often remaining sub clinical and resolving promptly at the end of CPB, in its most extreme form this inflammatory response may be associated with the development of the systemic inflammatory response syndrome (SIRS) that can often lead to major organ dysfunction (MODs) and death. Here, we review the pathophysiology behind the development of this "whole body" inflammatory response and some of the methods currently used to minimise it. PMID- 17462275 TI - From frown lines to fissures: therapeutic uses for botulinum toxin. AB - INTRODUCTION: Since the pharmacological mode of action of botulinum toxin (BTX) has been elucidated, its therapeutic potential has been increasingly recognised. The aims of this review were to summarize our current understanding of the pharmacological action of this agent and to review its therapeutic uses. METHODS: An electronic literature search with Medline (January 1965 to December 2004) was carried out to identify articles related to the pharmacological mode of action and clinical uses for botulinum toxin using the keyword "botulinum toxin". RESULTS AND CONCLUSION: Botulinum toxin A is emerging as a valuable clinical tool, both for diagnostic and therapeutic purposes in a wide variety of disorders, and is already the treatment of choice for selected conditions. Better understanding of its modes of action may identify alternative targets for pharmacological intervention, and may allow development of longer acting drugs with lower immunogenicity. Therapeutic uses of BTX-A must be assessed systematically in prospective studies, and the clinical role of other subtypes requires evaluation. PMID- 17462276 TI - Frequency volume charts should be used in men with lower urinary tract symptoms. AB - A frequency volume chart is a simple, easy-to-use, non-invasive tool that is useful in the assessment of patients with lower urinary tract symptoms. Though more sophisticated techniques are now available for diagnosis, the frequency volume chart should still be considered the first line investigation. In this review we summarize the indications and value of this age old investigation. Urologists and primary care physicians should use the frequency volume chart more frequently in their practice. PMID- 17462277 TI - The teaching of surgery in the undergraduate curriculum. Part II--Importance and recommendations for change. AB - In the past decade, the teaching of surgery in the undergraduate curriculum has undergone considerable changes in quantity, mode and method of delivery. This is a result of the radical reforms of higher education, the health service and the undergraduate medical curriculum. The changes are complex and require us to ask the questions: how important is the teaching of surgery in the modern medical undergraduate curriculum and is there a need for change? We aim to tackle these questions and propose practical action which medical schools can take to ensure that they deliver effective surgical teaching within the modern medical curriculum and health service. PMID- 17462278 TI - Hogarth's 'Gin Lane' and 'Beer Street'. AB - The original intention of this paper was to draw on the link between art and medicine through the appreciation of a pair of prints created by Hogarth in 1750. This was done firstly, by viewing them as a direct health promotional campaign to curb alcoholism, and secondly, by studying Hogarth's use of the depiction of disease as a means to achieve this. However, in analysing his motivation another more permanent link, still resonant in today's society, was revealed--that between health and social structure. PMID- 17462280 TI - The editor's diary: semantics can be bad for your health! PMID- 17462281 TI - Battlefield surgery 2005. PMID- 17462282 TI - Does surgery induce angiogenesis in breast cancer? Indirect evidence from relapse pattern and mammography paradox. AB - A significant bimodal relapse hazard pattern has been observed in two independent databases for patients untreated with adjuvant chemotherapy. This implies there is more than one mode of relapse. The earliest and most closely grouped relapses occur 8-10 months after surgery for young women with node-positive disease. Analysis of these data using computer simulation suggested that surgery probably instigated angiogenesis in dormant distant disease in approximately 20% of cases for premenopausal node-positive patients. We explore if this could explain the mammography paradox for women aged 40-49: an unexplained temporary excess in mortality for the screened population compared to controls. Calculations based on our data predict surgery-induced angiogenesis would accelerate disease by a median of two years and produce 0.11 early deaths per 1000 screened young women in the third year of screening. The predicted timing as well as the magnitude of excess mortality agree with trial data. Surgery-induced angiogenesis could account for the mammography paradox for women aged 40-49 and the bimodal relapse hazard pattern. According to the proposed biology, removing tumors could remove the source of inhibitors of angiogenesis or growth factors could appear in response to surgical wounding. While this needs confirmation, this could be considered when designing treatment protocols particularly for young women with positive nodes. It reinforces the need for close coordination between surgical resection and ensuing medical intervention. Women need to be advised of risk of accelerated tumor growth and early relapse before giving informed consent for mammography. PMID- 17462283 TI - The surgical procedure and clinical results of the duct to mucosa pancreaticojejunostomies with resection of jejunal serosa. AB - BACKGROUND: Pancreatic anastomotic leakage often results in severe complications of sepsis, intra-abdominal bleeding, pancreatic fistula, and is a significant cause of morbidity and mortality. An appropriate technique to minimize pancreatic leakage is very important. Recently we have performed duct to mucosa pancreaticojejunostomy with resection of jejunal serosa and obtained positive results. PATIENTS AND METHODS: During 1999-2005, 52 patients (25 females, 27 males) underwent duct to mucosa pancreaticojejunostomy with resection of jejunal serosa after pancreatic head resections for benign (n=6) and malignant disease (n=46). The mean age was 64.0 years (range 33-80). RESULTS: Mean post-operative hospital stay was 32.3 days. Morbidity rate due to early post-operative complication was 7.7% (pulmonary embolism in 1, pneumothorax in 1, wound infection in 2), with no pancreatic leakage. CONCLUSIONS: There were low complication rates and an absence of pancreatic anastomotic leakage was observed in 52 patients. We consider that this pancreatic anastomotic technique is extremely favorable for pancreaticojejunostomy. PMID- 17462284 TI - A review of contemporary surgical alternatives to permanent colostomy. AB - OBJECTIVES: To review the options available to patients with faecal incontinence with failed conservative treatment and/or failed anal sphincter repair and assessing the current indications and results of these options. METHODS: A literature search of MEDLINE, EMBASE and Cochrane databases was performed using the relevant search terms. RESULTS: Continent options for patients with severe or end stage faecal incontinence include the creation of a form of an anal neosphincter and more recently sacral nerve stimulation. Over half the patients, who are candidates, may benefit from these procedures, although long term results of sacral nerve stimulation are unknown. Dynamic graciloplasty improves the continence in 44-79% of the patients. The complications include frequent reoperations, high incidence of infection and obstructive defaecation. The success rates of artificial bowel sphincter vary between 24% and 79%. Once functional, the artificial bowel sphincter seems to improve the continence in the majority of the patients. Device removal due to infection, obstructive defaecation and pain is a frequent problem. Sacral nerve stimulation is claimed to result in improvement in continence in 35-100% of patients. The main risks in this procedure are infection, electrode displacement and pain. CONCLUSIONS: All these procedures have high complication rates and have moderate success rates only. A major proportion of patients will need reoperations and hence high motivation is necessary for patients who undergo these procedures. A uniform standard for measurement of success is also necessary so that these procedures can be compared with each other. PMID- 17462285 TI - A review of the surgical management of metastatic gastrointestinal stromal tumours (GISTs) on imatinib mesylate (Glivec). AB - Gastrointestinal stromal tumours (GISTs) are defined as a group of C-KIT positive mesenchymal tumours of the gastrointestinal tract. Although they may arise throughout the gut, the commonest sites are stomach and small intestine. Over 80% of metastases are to the liver and omentum. Targeted therapy (imatinib) can inhibit C-KIT and thereby aberrant tumoural proliferation. Imatinib may induce shrinkage of lesions and cystic change. Such physical changes often correspond with reduced metabolic activity demonstrated by (18-FDG)PET scans. These changes may enable metastatectomy reducing tumour pain and the risk of haemorrhage and rupture in the short term. In the long term, resection may lessen the risk of recurrence by removing potentially resistant clones. The precise role of palliative resection for GIST metastases on imatinib remains unclear. Imatinib has changed the natural history of metastatic GISTs, with increased survival times. Surgery remains an important management strategy in the metastatic setting because complete pathological responses are rare with imatinib. Surgery is likely to provide the best palliation, greatest reduction in tumour burden and eliminate resistant clones. A multidisciplinary team approach with expertise concentrated in a few centres specialising in the management of these rare tumours is vital to the successful outcome. Future issues regarding the management of differential response of the metastases to imatinib are highlighted. With the emergence of techniques enabling identification of the precise mutational status of the C-KIT oncogene, the imatinib/surgery sequence could be tailored to the type of C-KIT mutation. PMID- 17462286 TI - Advances in the management of thyroid cancer. AB - The incidence of thyroid cancer is rapidly increasing in the United States. A large number of incidentalomas are found during routine head and neck evaluations. The diagnostic workup still revolves around fine needle aspiration biopsy. Ultrasound guided fine needle aspiration biopsy is likely to yield the best results. Surgical resection offers the best treatment choice. Controversy continues in relation to total versus less than total thyroidectomy. The incidence of complications is inversely proportional to the extent of surgery and obviously related to the experience of the operating surgeon. The decision regarding the extent of thyroidectomy should be based on prognostic factors and risk groups. Prognostic factors are well defined, such as age, grade of the tumor, extrathyroidal extension, size, distant metastasis, and histology. Nodal metastasis has minimal implications. Based on prognostic factors, thyroid cancer can be divided into low, intermediate and high risk groups. In the high risk group and in selected intermediate risk patients, radioactive iodine dosimetry and ablation should be considered after total thyroidectomy. PET scanning and the use of recombinant TSH have been major advances in follow-up care for patients with thyroid cancer. Thyroglobulin appears to be a very good tumor marker for follow-up. No major breakthrough is noted in the management of anaplastic thyroid cancer, however, identification of RET mutation has been extremely helpful in evaluating the family members of the patient with medullary thyroid cancer with strong consideration given to total thyroidectomy. PMID- 17462287 TI - An enlightened age: building the naval hospitals. AB - Summerson writes that, in the spirit of the Enlightenment, the notion of 'bienfaisance', literally meaning the desire to render society more reasonable and more humane, liberated the scope of both hospital and prison planning. Both types of institution housed people who were deprived of their freedom either by disability or by force of law; therefore, we find similarities in the disposition of space and in the degree of humanitarianism expressed in their outward appearance. This observation can be transferred to naval hospital design, where architecture was combined to fulfil a twofold purpose. The high walls, sturdy massing and pared-down details intimated a strict economy of means, but also a strong sense of gravitas and authority. This visual authoritarian character was fundamental to the identity of a naval hospital, for whilst the prime intention was to provide efficient medical care, an equally important consideration was to maintain discipline and prevent sailors from escaping. PMID- 17462289 TI - Distributive justice, starfish and natural disasters. PMID- 17462290 TI - Authors respond to controversy surrounding breast cancer study. PMID- 17462291 TI - Viewing the pyramid--through a glass, darkly? PMID- 17462292 TI - Nanotechnology, nanomedicine and nanosurgery. PMID- 17462293 TI - Quality of drainage blood: survival of red cells after re-transfusion and content of free hemoglobin and potassium. AB - Re-transfusion of drainage blood is widely used in orthopedic surgery, but objective evidence of the efficacy of re-transfusion of drainage blood in view of post-transfusion survival of RBC has not been given so far. With this study we wanted to evaluate the efficacy and safety of transfusion of drainage blood collected with HandyVac autotransfusion system. In 7 patients red cells in drainage blood were labeled with biotin and percentage of labeled red cells in circulation were determined immediately after re-transfusion, and during 10 days after surgery. To assess further unwanted side-effects of re-transfusion of drainage blood potassium and free hemoglobin were determined in the collected blood. Ten days after re-transfusion at mean 78.9% of drainage-blood derived RBC were found in circulation. Free hemoglobin in drainage blood ranged from 16.8 to 59.2 mg/dL; potassium in drainage blood ranged from 3.84 to 4.52 mmol/L. Our results suggest that re-transfusion of drainage blood collected with HandyVac autotransfusion system is an efficient procedure that seems to be safe in view of free hemoglobin and potassium in the product. PMID- 17462294 TI - Uses and abuses of digital imaging in plastic surgery. AB - BACKGROUND: Surgeons extensively rely on photographic communication for documenting surgical results, teaching and research, and obtaining informed consent from patients. With the advent of digital photography and widespread availability of sophisticated image manipulation software, the potential for committing digital fraud cannot be discounted. METHODS: Ten 'before' and 'after' plastic surgical photographs were selected, and a number of them were digitally enhanced using a standard desktop software by a non-expert in digital photography. A panel of 10 consultant plastic surgeons was asked to judge which, if any of the images had been digitally manipulated. RESULTS: Expert assessment had a sensitivity of only 12% in identifying digitally manipulated images. Furthermore, there was poor interobserver agreement with an Intraclass Correlation Coefficient of 0.39. CONCLUSION: Digital fraud is easy to commit and difficult to detect. Furthermore, a number of inadvertent and simple image manipulation functions can also amount to misrepresentation. There may be scope for cooperation within editorial circles to set standards for the submission of digital photographs. Surgeons need also to be aware of the potential for misrepresentation of information through digital image manipulation and exercise caution in the communication of digital photographic information. PMID- 17462295 TI - Comparison of electrotherapy of hemorrhoids and Ferguson hemorrhoidectomy in a randomized prospective study. AB - BACKGROUND: Ferguson hemorrhoidectomy has been shown to be associated with significant amount of post-operative (post op) pain and complications. However, electrotherapy in which hemorrhoidal tissue is not excised might not be associated with severe complications. OBJECTIVE: Our aim was to compare the results of Ferguson hemorrhoidectomy with electrotherapy methods using 16 and 30 mA (milliampers) direct current (DC). METHODS: Four hundred and eight patients with symptomatic hemorrhoids, grades 1, 2 and 3, were randomly assigned into 3 groups. Group A (136 patients) underwent Ferguson hemorrhoidectomy, group B1 (136 patients) and group B2 (136 patients) were subjected to electrotherapy using 16 and 30 mA, respectively. The groups were compared in terms of duration of procedures, duration of hospital stay, post op pain severity and post op complications including recurrence, infection and non-healing ulcers. RESULTS: All patients in group A had severe pain for 7-14 days of post op. However, in group B1, 88(65%) patients had mild pain during the treatment and 1st post op day; 28(21%) of them could not tolerate the operation; 20(15%) of them had mild pain and 10(7.5%) of them had moderate pain up to day 7. In group B2, 47(35%) of patients had sever pain for 6 h and 20(15%) of them experienced mild pain for 2-7 days post op. The one day hospital stay in group A and group B2 were 82 and 97%, respectively, while patients in group B1 were treated as out patients. Mean procedure time for one hemorrhoidectomy in group A was 23 min, in electrotherapy using 16 and 30 mA was 9.7 and 6.1 min, respectively. The overall success rate with the first application in group B1 was 57% and in group B2 was 93%. CONCLUSION: Electrotherapy method using 30 mA DC could significantly decrease post op pain, operation time and hospital stay. This method had good success rate and very low post op complications compared to Ferguson hemorrhoidectomy and using 16 mA method. Therefore, due to its effectiveness, less pain, rapidity and safeness, we recommend it. PMID- 17462296 TI - Space exploration--surgical insights and future perspectives. AB - NASA's space exploration initiative envisions a return to the moon by 2020, the construction of inhabited lunar bases and manned missions to Mars. Such an ambitious program harbours increased risks, both logistical and physical (particularly that of trauma) within the context of a microgravity environment. This paper also discusses the cellular response to microgravity and the potential scientific and technological benefits of Space exploration. PMID- 17462297 TI - An introduction to death receptors in apoptosis. AB - Apoptosis is a source of much research interest across many fields, including developmental biology, immunology and oncology. As the exact pathways of this process are identified, so too are potential avenues for therapeutic application. Death receptors are important in inducing apoptosis and together with their ligands have become a source of attention as potential therapeutic agents. This review provides an introduction to the role of death receptors in apoptosis, together with a look at possible areas where this information may be applied therapeutically. PMID- 17462298 TI - The risks and benefits of cyclo-oxygenase-2 inhibitors in prostate cancer: a review. AB - Cyclo-oxygenase (COX), also referred to as prostaglandin (PG) endoperoxidase synthase, is a key enzymatic mediator in the production of arachidonic acids to PGs and eicosanoids. Two isoforms of COX exist, namely COX-1 and COX-2, which have distinct physiological functions and tissue distribution. Epidemiological studies suggest that regular consumption of aspirin and/or other non-steroidal anti-inflammatory drugs (NSAIDs), which inhibit COX, could notably reduce the risk of developing many cancers. COX-2 expression has been shown to increase in many cancers and cancer cell lines, including human prostate adenocarcinoma. COX 2 may also be upregulated in proliferative inflammatory atrophy (PIA) of the prostate, a pre-neoplastic lesion. The COX-2 pathway may therefore be a useful target for chemoprevention of prostate cancer, and there is much interest in exploring this with the use of COX-2 inhibitor drugs such as celecoxib. While there is concern regarding the cardiovascular toxicities of coxibs, there is no evidence that there is any increased risk with the use of celecoxib in the short term neoadjuvant setting. PMID- 17462299 TI - Breast cancer surgery and angiogenesis: stem cell cycle may explain heterogeneity of recurrence. PMID- 17462300 TI - Breast cancer heterogeneity may explain peaks in recurrence. PMID- 17462301 TI - Bradford-Hill Criteria provide the way ahead for controversial theory. PMID- 17462302 TI - Does surgery induce angiogenesis in breast cancer? Supporting evidence from Africa. PMID- 17462303 TI - Surgery-induced angiogenesis. PMID- 17462304 TI - Cultural fears of early mortality already exist. PMID- 17462309 TI - Economic evaluation of MR cholangiopancreatography compared to diagnostic ERCP for the investigation of biliary tree obstruction. AB - BACKGROUND: Use of magnetic resonance cholangiopancreatography (MRCP) for confirmation of presence of biliary obstruction is virtually risk-free. However, unlike diagnostic endoscopic retrograde cholangiopancreatography (ERCP), no therapeutic option can be offered simultaneously with MRCP. The aim of the study is to assess the cost-effectiveness of MRCP when compared with the conventional practice of diagnostic ERCP for the investigation of biliary obstruction in adults. METHODS: Cost-effectiveness analysis from the perspective of the health care provider. Sensitivity analysis includes presentation of a family of cost effectiveness acceptability curves and the impact of different risks of common bile duct stones associated with ultrasound and liver function test results. The main outcome measure is cost per quality adjusted life year (QALY). RESULTS: Baseline results, at 37% probability of common bile duct stones, show that MRCP is the dominant strategy, with expected savings of 149 pounds sterling (325 pounds sterling to - 15 pounds sterling) and expected QALY gain of 0.011 (0 0.030) per case. The probability of avoiding unnecessary therapeutic ERCP is 30%. For patients at high risk of common bile duct stones (probability >60%) ERCP is the preferable strategy. CONCLUSIONS: The baseline estimate is that MRCP would be both cost saving and would result in improved quality of life outcomes compared to diagnostic ERCP, but its potential sources of economic benefit are highly dependent on access to, and waiting lists for adequate MRI technology at hospital level. PMID- 17462310 TI - Does the method of harvesting the saphenous vein for coronary artery bypass surgery affect venous smooth muscle cells? iNOS immunolabelling and ultrastructural findings. AB - OBJECTIVES: Coronary artery bypass surgery (CABG) using conventionally harvested saphenous vein (SV) as a graft is characterised by a high graft failure rate. It is believed that vein handling during harvesting is responsible for this, as damage to the vein's structure including its sources of endothelial nitric oxide synthase (eNOS) is apparent. Here we investigated the distribution of inducible nitric oxide synthase (iNOS) in conventionally harvested SV grafts and grafts prepared by a less-invasive 'no-touch' technique for CABG. Focus was on vascular smooth muscle cells (VSMCs). PATIENTS AND METHODS: Six patients undergoing CABG were selected for the study; their SVs were harvested as grafts by conventional and 'no-touch' techniques and subsequently examined using fluorescent and transmission electron microscopy combined with immunolabelling to detect iNOS and structural changes in the grafts at the time of implantation. RESULTS: The following were observed in conventionally harvested grafts: (i) damage to the VSMCs and (ii) induction of iNOS in these cells, (iii) heterogeneity of VSMCs-the presence of iNOS-positive and iNOS-negative VSMCs which were also (iv) in close contact with each other. In contrast, no damage to VSMCs and no expression of iNOS in these cells were observed in the 'no-touch' SV preparations. CONCLUSIONS: The harvesting procedure influences the structure of VSMCs and the expression of iNOS in SV graft at the time of implantation. Whether rapid stimulation of iNOS in VSMCs in conventional SV grafts has a protective or harmful effect on the graft patency remains to be determined. PMID- 17462311 TI - Evaluation outcome of replanted digits using the DASH score: review of 38 patients. AB - The ultimate usefulness of replanted fingers is related to the adequacy of nerve, tendon and bone repair. Thirty-eight patients with successful replantation of the thumb, a single finger or after multiple digital amputations were followed up clinically. The subjective disability after finger replantation was evaluated by means of the DASH score, and the presence of cold intolerance was assessed. The subjective outcome of the achieved replantation reached an overall DASH score of 12.3. Patients after thumb replantation reached a score of 10.0, after a single finger replantation DASH 11.2 and after multiple finger amputations and replantation of at least one finger, DASH 16.1. Cold intolerance was subjectively found in 86.7% of all hands with replanted fingers without a correlation to the patients DASH scores. The patient's evaluation of their limb function after replanted digits by means of the DASH score, when combined with an objective external assessment, represents a valuable comparative tool. PMID- 17462312 TI - Psychological responses and support needs of patients following head and neck cancer. AB - The patient with head and neck (H&N) cancer is prone to psychological distress immediately following diagnosis and during the treatment phase. Lowered mood is typical and tends to extend beyond the treatment phase. There is little evidence for a specific treatment method predicting a characteristic psychological response. Rather, patients' reactions vary widely according to fears of recurrence, health beliefs, personality, coping and available support. Patient reports of quality of life show a return to pre-treatment status after a year but are determined to some degree by initial depression levels and dispositional factors such as optimism. Information provided to patients (e.g. leaflets, booklets of written guidance) by specialist treatment centres about the disease and its management require sustained effort in their design and distribution. Our understanding of patient responses to this disease has improved and has assisted in the development of psychological interventions. Controlled trials will provide important evidence of the components, effects and sustainability of these experimental programmes, and improve overall care plans for this often neglected patient group. PMID- 17462313 TI - Treatment of intra-abdominal and skin and soft tissue infections: the role of the glycylcyclines. AB - The need for new, effective agents to treat multidrug-resistant infections continues to grow as more and more bacteria develop resistance that may result in clinical therapeutic failure. This is particularly true for common surgical infections, such as complicated intra-abdominal infections, which frequently involve multiple pathogens, making therapy with a broad-spectrum antibiotic an important treatment intervention, and also for complicated skin infections, which often involve methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin resistant enterococci (VRE). 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: linezolid, quinupristin dalfopristin and most recently daptomycin have all been FDA-approved for the treatment of skin and skin structure infections. This review examines the potential role of a new class of investigational agents, the glycylcyclines, also recently FDA-approved and currently under review for European licensing, in the treatment of complicated skin infections and intra-abdominal infections. Tigecycline, the first of the glycylcyclines, has shown excellent activity in Phase III studies of these infections, achieving clinical success rates ranging from 70% to 91%. Furthermore, it has a good safety profile, suggesting it will be a clinical useful addition to current therapeutic options for the treatment of complicated skin infections and intra-abdominal infections. PMID- 17462314 TI - Hand hygiene: an evidence-based review for surgeons. AB - This review of the literature discusses the scientific evidence behind using different hand hygiene agents on the surgical ward, and in theatre for preoperative disinfection. It considers the mechanism of action of the agents and their effectiveness against different pathogens, as well as possible future agents, and how they are tested. It addresses problems such as the poor compliance with hand hygiene guidelines by healthcare workers (especially doctors) and investigates what can be done to improve compliance. Finally, it demonstrates the reduction in hospital acquired infection (HAI) rate that can be achieved by improving hand hygiene compliance, and shows that the savings associated with this easily outweigh the cost. PMID- 17462315 TI - Goya: 'in sickness and in health'. AB - The following paper serves primarily to discuss the nature and cause of the multiple, documented illnesses suffered by the 18th Century Spanish artist Francisco Goya y Lucientes. In doing so it explores the artist's defiance of contemporary social mores, religious doctrines and the rigid standards of artistic romanticism. Furthermore, it examines the impact of illness upon artistic expression and the apparent correlation between heightened artistic proficiency and clinical depression with particular reference to the aforementioned artist. In addition, it provides a brief insight into the complexion of medical practice in Spain during the period of the Inquisition contrasted with those standards of conduct encompassed within the newly emerging movement of Enlightenment. PMID- 17462316 TI - Perspectives on this issue of the IJS. PMID- 17462317 TI - Mammographic screening: a common cause of statistics! PMID- 17462318 TI - The IJS and the COPE seminar 2006. PMID- 17462319 TI - Intellectual property rights and living organisms. PMID- 17462320 TI - The IVF-stem cell interface. PMID- 17462321 TI - Ethical considerations related to organ transplantation and Islamic Law. AB - With the increasing number of transplantable organs and tissues, as well as improvements in transplantation results, has come a severe shortage of organ donors. Organs for transplantation are usually obtained from living genetically related donors or from heart-beating cadavers. Unfortunately, these sources have so far been unable to keep up with demand. As a result, there is a large and steadily increasing number of potential recipients awaiting transplantation, some of who will die before an organ can be found. These trends have raised many ethical, moral, societal and in particular religious (Islamic Law) issues regarding supply, the methods of organ allocation, and use of living donors. Several ethical dilemmas regarding case selection, allocation within the law, medical problems, and economic sources have now to be confronted. Despite this, the legal framework regulating transplantation in Iran was recently enhanced in comparison to other Islamic countries. PMID- 17462322 TI - Facial transplantation: a real option in facial reconstruction? PMID- 17462323 TI - Prospective evaluation of patients with acute cholecystitis treated with percutaneous cholecystostomy and interval laparoscopic cholecystectomy. AB - BACKGROUND: Emergency laparoscopic cholecystectomy has been advocated for the treatment of acute cholecystitis; however it can be a difficult task, especially in public hospitals, with relatively high conversion and complication rates. Percutaneous cholecystostomy is a simple and effective procedure allowing patients to recover from the acute event and undergo elective laparoscopic surgery at a later stage. METHODS: We prospectively assessed a protocol of initial conservative treatment in patients admitted with acute cholecystitis. Patients who did not respond to medical treatment were treated by percutaneous cholecystostomy. Following discharge the patients were seen in the outpatient clinic and elective laparoscopic cholecystectomy was considered and scheduled as necessary. The details of the operation were collected with emphasis on complications and conversion rate. RESULTS: During a 3-year period, 224 patients who were admitted with acute calculous cholecystitis entered the protocol. Fifty four patients did not improve under medical treatment and percutaneous cholecystostomy was performed. In spite of adequate drainage, 5 patients still did not improve: 3 patients were successfully operated upon urgently and recovered, while 2 patients who had severe concomitant diseases and multi-organ failure, died. Forty-nine patients were discharged with the catheter and later re evaluated for elective operation. In 7 patients common bile duct stones were found and were removed by ERCP prior to the elective operation. Twenty-five patients underwent delayed laparoscopic cholecystectomy with a low conversion rate (8%), and only minor complications (16%). CONCLUSIONS: Conservative treatment and delayed operation is still an acceptable choice in the treatment of acute cholecystitis. Percutaneous cholecystostomy is an effective tool, with high success rate and low morbidity, and allows for better pre-operative evaluation of the biliary system and safe interval laparoscopic surgery. Laparoscopic cholecystectomy after drainage of the gall bladder is a low morbidity procedure with relatively low conversion rate. PMID- 17462324 TI - Breast conservation treatment in women with locally advanced breast cancer - experience from a single centre. AB - INTRODUCTION: In absence of randomized evidence to support safety of conservative surgery (BCT) in locally advanced breast cancer (LABC), we analyzed a cohort of 664 women with LABC treated during January 1998 to December 2002 at Tata Memorial Hospital, Mumbai, India. MATERIALS AND METHODS: All were treated with a multimodality regimen comprising of neoadjuvant chemotherapy (NACT) followed by surgery (modified radical mastectomy or BCT) and adjuvant radiotherapy and hormone therapy. The outcome was evaluated to assess safety of BCT. RESULTS: 71% (469/664) women responded to NACT (22% clinical CR and 49% PR) and 28.3% (188/664) underwent BCT. Positive lumpectomy margins were reported in 8.5%, with gross presence of tumor at the margins in 2.3% requiring a revision surgery. At a median follow-up of 30months, local relapse rate was 8% after BCT and 10.7% after mastectomy. The 3-year local DFS was better post-conservation than after mastectomy (87% vs 78%, P=0.02). The disease-free survival (DFS) was also superior after BCT, 72% vs 52% (P<0.001) at 3years and 62% vs 37% (P<0.001) at 5years respectively. On multivariate analysis, presence of lymphatic vascular emboli (LVE) was the major significant predictor of local recurrence (P<0.001, HR 2.52, 95% CI 1.52-4.18). DFS was better after BCT [(P<0.001, HR 2.0 (95% CI 1.38 2.91)]; shorter DFS was noted in LVE positive (HR 1.54, P=0.007) and larger residual disease after NACT (HR 1.13, P=0.001). CONCLUSION: BCT is technically feasible and safe post neo-adjuvant chemotherapy in women with LABC with no detriment in outcome. PMID- 17462325 TI - Early management of atrial fibrillation in general surgical in-patients. AB - BACKGROUND: It has been suggested that new-onset atrial fibrillation (AF) in non cardiac surgical patients should trigger a thorough search for other morbidity. We reviewed our unit's management of new-onset AF to determine whether this target is achieved. METHODS: Patients under the care of a general surgeon who developed new-onset AF during their inpatient stay were identified from a prospectively maintained database of surgical in-patients. Their case-notes were reviewed to determine whether a precipitating cause for the AF was sought or identified. RESULTS: Thirty-one patients developed new-onset AF. Almost half had positive findings on cardiovascular or respiratory system examination and assessment. However, 35% of patients had no respiratory examination, 58% had no cardiovascular examination and 55% had no abdominal examination performed. Eighty one percent had another underlying complication diagnosed within 24h of the AF. Twenty-six percent had an intra-abdominal collection. Of those with an underlying complication, 52% were not diagnosed at the time of initial assessment for AF. Twenty percent of patients died within 30 days of the AF. CONCLUSIONS: New-onset AF in general surgical patients is associated with considerable morbidity. A thorough clinical evaluation and early involvement of senior surgical staff are recommended. PMID- 17462326 TI - Surface disinfection of packed red blood cells with 70% ethanol. AB - No data or recommendations are available on feasibility of surface disinfection of blood bags, but some circumstances can make such procedures inevitable. Impact of immersion of blood bags in 70% ethanol for 30min was investigated with respect to alcohol penetration and changes of hemolysis parameters in the product, and bag material changes influencing material stability and composition. After immersion ethanol concentration in blood bags was below detection limit. Hemolysis parameters did not differ between blood products that had been exposed to ethanol and a control group. Inner surface of the bag material was unchanged according to our infrared spectrometry results. Also endurance testing showed no altered results. We conclude that immersion of blood bags in 70% ethanol for surface disinfection is a safe procedure for the quality of the blood product and the bag material. PMID- 17462327 TI - Deceit and fraud in medical research. AB - Deceit and fraud in medical research is a serious problem for the credibility of published literature. Although estimating its prevalence is difficult, reported incidences are alarming. The spectrum of the problem ranges from what may seem as rather innocuous gift authorship to wholesale fabrication of data. Potential factors which may have promoted fraud and deceit include financial gain, personal fame, the competitive scientific environment and scientific hubris. Fraud and deceit are difficult to detect and are generally brought to the fore by whistleblowers. Although most cases may be dealt with at an institutional level, regulatory organisations such as the Committee on Publication Ethics (COPE) and Medical Research Ethics Committee (MREC) have been established to monitor and try to remedy the problem. PMID- 17462328 TI - The management of rectal cancer in a resource poor environment - a review. AB - Rectal cancer is an increasing problem in the developing world. There is little written on how to manage this problem outside the confines of major teaching hospitals in Western Countries. In these centres debate surrounds preoperative staging, the use of preoperative radiotherapy and sophisticated sphincter preserving procedures. The literature is complex and of little relevance to those faced with a patient with rectal cancer in rural Africa or Asia far from the ivory towers. This review aims to combine the best of evidence based medical practice related to the management of rectal cancer with the practical realities of operating in resource poor environments. In this situation staging is by means of simple radiology and a clinical examination supplemented by an examination under anaesthetic. If there are no distant metastases and the tumour is freely mobile a resection can be attempted. An abdominoperineal resection is a good operation with a proven track record. If an anastomosis can be fashioned then an anterior resection is an excellent operation and should be performed extrafascially to avoid local recurrence. It is vital to counsel the patient preoperatively. The pros and cons of referral to a centre of excellence need to be discussed with the patient prior to any intervention. PMID- 17462329 TI - Deceit and fraud in medical research - education is the way forward. PMID- 17462330 TI - Deceit and fraud in medical research - publish or perish culture is to blame. PMID- 17462331 TI - The IVF-stem cell interface - public opinion and religious views matter. PMID- 17462333 TI - Perspectives on this issue of the IJS. PMID- 17462332 TI - Deceit and fraud in medical research - intermediary, commercially based research teams are to blame. PMID- 17462334 TI - Introducing the IJS.com: transforming an international audience into a global community. PMID- 17462335 TI - Introducing Wikisurgery.com: the blueprint for a surgical architecture of participation. PMID- 17462336 TI - Pre-implantation genetic diagnosis (PGD): the spectre of eugenics or a "no brainer". PMID- 17462337 TI - Obesity and rationing within the NHS: joint replacement surgery. PMID- 17462338 TI - Should obese patients be denied knee surgery? A patient perspective. Maintenance of the machine--measurement and values. PMID- 17462339 TI - Controlled cardiac reoxygenation does not improve myocardial function following global myocardial ischemia. AB - BACKGROUND: It has been shown that abrupt re-exposure of ischemic myocardium to oxygen can lead to increased peroxidative damage to myocytes (oxygen paradox). Controlled cardiac reoxygenation, as an adjunct to substrate-enhanced cardioplegia, has been shown to improve myocardial function and limit reperfusion injury when utilizing standardized hyperoxic cardiopulmonary bypass (CPB). The objective of our study was to evaluate the effect of controlled reoxygenation on myocardial function following global ischemia employing normoxic CPB. STUDY DESIGN: Nineteen female swine (30-40kg) were placed on vented, normoxic CPB. They were subjected to 45-50min of unprotected global ischemia (aortic cross clamping) followed by 30min of controlled cardiac reperfusion utilizing substrate-enhanced cardioplegia. Group 1 maintained normoxic pO(2) (O(2) tension of 90-110mmHg). In Group 2, reoxygenation was titrated gradually and increased from venous to arterial levels (O(2) tensions from 40 to 110mmHg over 15min). We measured coronary sinus blood samples for CK, CK-MB, nitric oxide, and conjugated dienes at baseline, 5min into the cardioplegic resuscitation, 5min after the cross clamp removal, and just prior to the termination of the study. Hearts were pathologically studied and scored for evidence of tissue peroxidation. RESULTS: Although not significantly different, Group 1 (normoxic reperfusion) animals were more likely to wean from CPB (p=0.141) and had a higher mean arterial pressure (p=0.556). In Group 1, conjugated dienes were significantly higher 5min into the resuscitative protocol (p=0.018) and at the termination of bypass (p=0.035). Five of six animals in Group 1 eventually attained normal sinus rhythm as opposed to three out of 13 in Group 2 (p=0.041). There was no significant difference in histology scoring between the two groups for tissue peroxidation. CONCLUSION: This study of controlled cardiac reoxygenation in a lethal ischemic swine model failed to demonstrate that the use of controlled reoxygenation on the myocardial function following global ischemia was better with maintained normoxic pO(2) (with O(2) tensions of 90-110mmHg) than when reoxygenation was titrated gradually and increased from venous to arterial levels (O(2) tensions from 40 to 110mmHg over 15min). PMID- 17462340 TI - Vascular endothelial growth factor augments the healing of demineralized bone matrix grafts. AB - AIM: To produce a totally allogeneic graft material that would eliminate the need for harvesting the autogenous bone from patients. MATERIALS AND METHODS: Thirty six (10x5mm) defects were created in the parietal bones of 18 rabbits. Two experimental groups of 12 defects each were grafted with DBM(IM) alone and VEGF+DBM(IM), respectively. The remaining 12 defects were left empty as controls, and sacrificed on days 14, 30 and 60. Two rabbits, with four defects, of each group were sacrificed on day 14, 30 and 60 post grafting. The defects were retrieved for histological evaluation and quantitative analysis. RESULTS: No bone formed across the defect in the controls. The difference in the amount of new bone formation between experimental groups was significant especially 30 and 60days post grafting (p<0.0001), which were 149% and 100% respectively. CONCLUSION: VEGF+DBM(IM) powder is a promising graft material, and warrants further clinical investigation. PMID- 17462341 TI - In vivo laparoscopic robotics. AB - Robotic laparoscopic surgery is evolving to include in vivo robotic assistants. The impetus for the development of this technology is to provide surgeons with additional viewpoints and unconstrained manipulators that improve safety and reduce patient trauma. A family of these robots have been developed to provide vision and task assistance. Fixed-base and mobile robots have been designed and tested in animal models with much success. A cholecystectomy, prostatectomy, and nephrectomy have all been performed with the assistance of these robots. These early successful tests show how in vivo laparoscopic robotics may be part of the next advancement in surgical technology. PMID- 17462342 TI - Late follow-up of patients submitted to subtotal splenectomy. AB - BACKGROUND: Over the past 21years, we have performed more than 200 subtotal splenectomies, in which the upper splenic pole vascularized only by the gastrosplenic pole vascularized only by the gastrosplenic vessels is preserved, to treat different pathologic conditions. A meticulous follow-up of the postoperative results of this procedure is of fundamental importance. METHODS: All patients undergoing subtotal splenectomy were invited to be reviewed. A total of 86 patients who had undergone surgery 1-20years ago were gathered; the surgical procedure was performed for one of the following conditions: portal hypertension due to schistosomiasis (n=43), trauma (n=31), Gaucher's disease (n=4), myeloid hepatosplenomegaly due to myelofibrosis (n=3), splenomegalic retarded growth and sexual development (n=2), severe pain due to splenic ischemia (n=2) and pancreatic cystadenoma (n=1). Patients underwent a hematological examination, an immunological assessment, abdominal ultrasonography, computed tomography, scintigraphy and endoscopy. RESULTS: Increased white blood cell count and platelets were the only hematological abnormalities. No immunological deficit was found. Esophageal varices were still present in patients who underwent surgery because of portal hypertension although without rebleeding. The ultrasound, tomography and scintigraphy examinations confirmed the presence of functional splenic remnants without significant size alteration. CONCLUSIONS: Subtotal splenectomy seems to be a safe procedure that can be useful in treating conditions involving the spleen. The functions of the splenic remnants are preserved during long periods of time. PMID- 17462343 TI - Collection and storage of leukocyte depleted whole blood in autologous blood predeposit in elective surgery programs. AB - INTRODUCTION: The aim of this investigation was to provide evidence that leukocyte depleted whole blood meets the requirements for transfusion of the European Council and thus may be an alternative to leukocyte and plasma depleted packed red blood cells in autologous blood predeposit for patients undergoing elective surgery programs. MATERIAL AND METHODS: Standard units of 450mL blood were collected from 25 healthy male volunteers. Leukocyte depletion was done via inline filtration 4h after collection. Storage lesion was assessed by measuring the release of K(+), LDH, free hemoglobin, and lactate into the storage medium, as well as by the increase of hemolysis, the decrease of pH and consumption of glucose over a storage period of 35 days. As surrogate marker for red cell quality the intracellular concentrations of adenine nucleotides [ATP, ADP, AMP] were determined. RESULTS: The extent of storage lesion remained within the ranges of standard liquid storage conditions. Hemolysis was far below the threshold of 0.8% in all WB units at the end of their shelf life. Only minor changes of intracellular adenine nucleotide levels were measured indicating a preserved function of red blood cells in leukocyte depleted whole blood. At the end of shelf life 70%+/-18% of initial ATP levels were detected. CONCLUSION: Based on our data we propose that leukocyte depleted whole blood, stored for 35 days can be an option in the autologous blood supply as it meets the requirements for transfusion of the European Council. PMID- 17462344 TI - Diaphragm disease of the ileum. AB - Diaphragm disease is a condition wherein the bowel lumen is divided into a series of short compartments by circular membranes of mucosa and submucosa, leading to a pinhole lumen and subsequent obstruction. It is a very rare condition commonly attributed to non-steroidal anti-inflammatory drugs (NSAIDs) usage. We present two cases of obstruction secondary to diaphragm disease of the ileum diagnosed on histology of the resected specimen. One patient had no history of chronic NSAID intake. PMID- 17462345 TI - The magnificent seven: a history of modern thyroid surgery. AB - In the mid-19th century, thyroid surgery was regarded as a "proceeding by no means to be thought of" in Britain, "foolhardy" in Europe, and "horrid butchery" in America. By the mid-20th century, thyroid surgery was amongst the most efficient and safest of operations. This transition of modern thyroid surgery over a century was brought about by seven surgeons. The lessons of Theodor Billroth, Theodor Kocher, William Halsted, Charles Mayo, George Crile, Frank Lahey and Thomas Dunhill are to be forever relearned and respected. Let us remember this pantheon of pioneers, these "Magnificent Seven". PMID- 17462347 TI - Probabilistic economic model to compare MRCP with ERCP for the investigation of biliary obstruction is never complete but an on-going program. PMID- 17462348 TI - Surgical issues in early management of atrial fibrillation in general surgical in patients. PMID- 17462349 TI - Perspectives on this issue of the IJS. PMID- 17462350 TI - Valedictory editorial for the IJS. PMID- 17462351 TI - Metastasis to the breast: a rare site for secondary malignancy. AB - Metastases to the breast from extramammary neoplasms are rare. Few cases of melanoma metastatic to the breast have been described in the literature. Correct identification of metastatic breast malignancy is of great significance since this presentation may be confused with primary benign or malignant breast disease. PMID- 17462352 TI - Patterns of usage of complementary and alternative medicine in general surgical patients. AB - BACKGROUND: Use of complementary and alternative medicine (CAM) is becoming increasingly common. The purpose of this study was to determine the prevalence among general surgical patients, whether its use is related to disease type, and if postoperative patient-perceived problems and actual complications may be related to some CAM use. METHODS: One hundred fifty-one consecutive patients over a 3 month period were queried. Demographic information was obtained from the medical record pertaining to age, gender, race, marital status, treatment and postoperative complications. CAM practices were divided into three categories: body/structure, herbal medications/supplements, and mind/spirit. Data were analyzed for overall use of CAM and type of CAM. Relationships of CAM use to gender, age, race, and disease type, patient-perceived postoperative problems, and actual incidence of postoperative complications were determined. RESULTS: Sixty patients (40%) used some type of CAM: 17% body/structure interventions, 31% herbal medications, and 3% mind/spirit practices. Demographics of CAM use: 47% of Caucasians, compared to 29% of African-Americans (OR=2.2, p=0.03); 44% of females, compared to 34% of males (p=NS); 49% of patients60 years old (OR=2.0, p=0.04); 48% of patients with a cancer diagnosis, compared to 31% of patients with a benign diagnosis (OR=3.1, p=0.04). Thirty-three of CAM patients reported some type of postoperative problem, compared to 26% of non-CAM patients (p=NS). Actual complication rates were 8% for CAM patients, compared to 11% for non-CAM patients (p=NS). CONCLUSIONS: Use of CAM is relatively common, with younger, Caucasian patients with malignancies being the most common users. However, there seems to be no difference in perceived postoperative problems, nor actual postoperative complications between CAM and non-CAM users. PMID- 17462353 TI - Coronary artery bypass graft surgery outcomes among African-Americans and Caucasian patients. AB - There have been few studies to date that investigate the effect of race on outcomes related to coronary artery bypass grafting. The objective of the present study was to investigate race as an independent predictor of outcomes among patients undergoing coronary artery bypass graft (CABG). A nested case-control study from a twelve-year hospitalization cohort (N=9671) in which data were collected prospectively was conducted. Cases were African-American patients undergoing CABG (N=644). Controls were randomly selected Caucasian patients undergoing CABG (N=1932). Controls were matched to cases 3:1 on year of surgery. Fifteen preoperative and intraoperative risk factors and 14 outcomes were examined. The 14 outcomes of interest were length of stay, readmission to ICU, total ICU stay, total hours on ventilator post-op, reoperation for bleeding/tamponade, deep sternal wound infection, neurological complications, pneumonia, other pulmonary complications, renal failure, gastrointestinal complications, atrial fibrillation requiring treatment, in-hospital mortality, and intraoperative complications. Regression analysis was used to control for risk factors. Multivariate analysis revealed African-Americans were at greater risk for renal complications (OR 1.88, 95% CI 1.27-2.77), neurological complications (OR 1.34, 95% CI 1.01-1.77), and pulmonary complications (OR 2.11, 95% CI 1.72-2.59). African Americans had a significantly longer hospitalization post-operatively (OR 0.79, 95% CI 0.66-0.96), but were less likely to experience post-operative atrial fibrillation requiring treatment than Caucasians (OR 0.64, 95% CI 0.49-0.84). Even after multiple adjustments, African-Americans undergoing CABG surgery had significantly greater morbidity compared to Caucasian patients. PMID- 17462354 TI - Survival rates in dysvascular lower limb amputees. AB - BACKGROUND: To assess the 5-year and 10-year survival rates of major (above ankle disarticulation level) dysvascular lower limb amputees attending a sub-regional Disablement Services Centre (DSC) specialising in amputee rehabilitation. Also to investigate the association between survival rates, cause of dysvascularity, level of amputation, smoking status and occupational status. SETTING: The study was undertaken in sub-regional DSC for amputee rehabilitation covering a base population of about 3.5million people. Over 80% of lower limb amputations were done for dysvascularity (peripheral vascular disease, diabetic or combination). All these patients were followed up in the DSC for their prosthetic/amputee rehabilitation. Modular case records of 201 consecutive patients from 1994 to 1995 who had diagnosis of dysvascularity as the cause of major lower limb amputations, were scrutinised regarding their 10-year survival; demographic details, level of lower limb amputations, Above Knee (AK=Transfemoral), Below Knee (BK=Transtibial), smoking status, occupational status, healing of the stump at first assessment, cause of amputation and association of these factors with survival rates. RESULTS: Of 201 individuals with either AK or BK amputations, 60% (121) had AK amputations and 67% (134) were males, the mean age was 69years of age. Sixty-seven percent (97) had history of smoking, either current 43% (62) or prior 24% (35) smoking, and 59% (68) were skilled or non-skilled manual workers. Fifty-one percent (99) had diagnosis of peripheral vascular disease, whilst 34% (65) had combination of peripheral vascular disease and diabetes, diabetes on its own in 4% (7). In 12% (23) other causes were noted such as embolism, acute ischaemia, venous ulcers, etc. Regarding stumps healing at first assessment, healing was noted in 54% (109) whilst stump was unhealed in 46% (92). The median survival was 48months. Using Cox proportional hazards regression to identify association with survival, the hazard ratio (HR) was significant regarding level of amputation: HR 2.34; 95% confidence interval (CI) (1.58, 3.47), P<0.001 (a recent BK amputation increases the risk by 2.3 compared to a recent AK amputation in diabetic cohort and also in the peripheral vascular disease/diabetes cohort). Hazard ratio was less than 1.0 in bilateral amputees: HR 0.35, 95% CI (0.21, 0.60), P<0.001 (bilateral amputation decreases risk by 0.35). CONCLUSION: Our study indicates that the median survival remains at 4years, which is similar to the previously published evidence in the Finish study of 1998 and the earlier study from Scotland in 1992. Unlike the previous data, our study indicates that patients with BK amputations have a higher hazard ratio than the AK amputees, and an association with diabetes has poorer prognosis regarding survival. PMID- 17462355 TI - Effects of diclofenac sodium on bursting pressures of anastomoses and hydroxyproline contents of perianastomotic tissues in a laboratory study. AB - BACKGROUND: Non-steroidal anti-inflammatory drugs are commonly used to relieve postoperative pain. Diclofenac sodium is a non-steroidal anti-inflammatory drug used as an analgesic during the postoperative period. Diclofenac sodium has powerful anti-inflammatory actions. As anti-inflammatory action may interfere with wound healing, we have studied the effects of diclofenac sodium on the healing of colonic anastomosis in rats. METHODS: Thirty-six Sprague-Dawley rats were used. Resection and anastomosis was performed on the distal colon of rats. The diclofenac group of rats received 4mg/kg/day diclofenac sodium intramuscularly. The control group received 0.1mL/day 0.9% NaCl intramuscularly. Anastomoses were evaluated with bursting pressure of the anastomoses and hydroxyproline level of the perianastomotic tissues on the third and seventh postoperative days. The Mann-Whitney U-test was used for statistical analysis. RESULTS: Three rats from each group died because of septic complications due to anastomotic leaks. Bursting pressures of the diclofenac group of rats (3rd day: 40.12+/-6.72mmHg; 7th day: 172.02+/-30.74mmHg) were lower than controls (3rd day: 54.13+/-8.11mmHg; 7th day: 206.00+/-27.14mmHg) both on the third and seventh days. Mean hydroxyproline contents of the perianastomotic tissues of diclofenac treated rats (3rd day: 1.97+/-0.17mug/mg tissue; 7th day: 3.61+/-0.32mug/mg tissue) were lower than controls (3rd day: 2.16+/-0.12mug/mg tissue; 7th day: 4.95+/-0.64mug/mg tissue) both on the 3rd and 7th days, similar to bursting pressures. CONCLUSIONS: Diclofenac sodium had a negative effect on bursting pressures of the colonic anastomoses and hydroxyproline contents of perianastomotic tissues of rats in our study. Anastomotic leak rates were not affected. The effects of diclofenac sodium should be studied in detail. PMID- 17462356 TI - The comparison between drainage, delayed excision and primary closure with excision and secondary healing in management of pilonidal abscess. AB - OBJECTIVE: There are numerous methods of treatment for pilonidal abscess; however the best method in acute pilonidal abscess has remained controversial. The present study was designed to compare drainage, delayed excision and primary closure with excision and secondary healing in this relation. METHODS: In a randomized clinical trial study among 102 patients with definite diagnosis of pilonidal abscess referred to the Colorectal Clinic of Nemazee Hospital, 80 patients who fulfilled the criteria of entering the study were selected and divided into two groups of A and B. The exclusion criteria were any history of pilonidal abscess operation, diabetes mellitus, renal failure and immunosuppression. In group A, drainage and delayed excision (3weeks afterwards) and primary closure were performed while in group B, excision and secondary healing was performed. Patients were followed twice a week for 1month and then 2, 6 and 12months after the operation. The two methods were compared in terms of time period for wound healing, postoperative complications and any sign of recurrence. RESULTS: Symptoms were relieved in all patients. All patients returned to work 7-9days after the operation. After 6months, there was no signs of recurrence in both groups. After 12months in group B, the same results were observed as previous months while in group A, 14% developed recurrence of pilonidal abscess. Wound infection was noticed in 5.6% of patients in group A and 2.5% in group B and the difference was not significant (p>0.05). All patients in group B developed wound healing during 6weeks except two of them who had a delay in this finding up to 8weeks. CONCLUSION: The results of drainage and primary wound closure were comparable to the excision and secondary wound healing except in the rate of recurrence which was more frequent in the primary wound closure. As a result, in spite of much property and comfort of primary wound closure, this method would not be recommended for all cases with acute pilonidal abscess. PMID- 17462357 TI - Clinical significance of microembolus detection by transcranial Doppler sonography in cardiovascular clinical conditions. AB - Transcranial Doppler can detect microembolic signals, which are characterized by unidirectional high intensity increase, short duration, and random occurrence, producing a "whistling" sound. Microembolic signals have been proven to represent solid or gaseous particles within the blood flow. Microemboli have been detected in a number of clinical cardiovascular settings: carotid artery stenosis, aortic arch plaques, atrial fibrillation, myocardial infarction, prosthetic heart valves, patent foramen ovale, valvular stenosis, during invasive procedures (angiography, percutaneous transluminal angioplasty) and surgery (carotid, cardiopulmonary bypass). Despite numerous studies performed so far, clinical significance of microembolic signals is still unclear. This article provides an overview of the development and current state of technical and clinical aspects of microembolus detection. PMID- 17462358 TI - Surgical repair of vaginal prolapse: a gynaecological hernia. AB - Female pelvic organ prolapse refers to the descent of the pelvic organs towards or through the vagina. The similarities between vaginal prolapse and herniae in their aetiology and treatment make this an interesting area for all those operating in the pelvis. It is a common condition with prevalence estimates varying from 2% for symptomatic prolapse to 50% for asymptomatic prolapse [Samuelsson EC, Arne Victor FT, Tibblin G, Svardsudd KF. Signs of genital prolapse in a Swedish population 20 to 59 years of age and possible related factors. Am J Obstet Gynecol 1999;180:299-305]. Approximately 50% of parous women will have some degree and only 10-20% of these seek medical help [Beck RP. Pelvic relaxation prolapse. In: Kase NG, Weingold AB, editors. Principles and practice of clinical gynecology. New York: John Wiley; 1983. p. 677-85]. The lifetime risk for surgery for prolapse has been estimated to be around 11.1%, and 30% will undergo re-operation for recurrent prolapse [Olsen AL, Smith VJ, Bergstrom JO, Colling JC, Clark AL. Epidemiology of surgically managed pelvic organ prolapsed and urinary incontinence. Obstet Gynecol 1997;89:501-6]. The aetiology of prolapse is multifactorial. Advancing age, parity and collagen weakness are all quoted as significant predisposing factors [Olsen AL, Smith VJ, Bergstrom JO, Colling JC, Clark AL. Epidemiology of surgically managed pelvic organ prolapsed and urinary incontinence. Obstet Gynecol 1997;89:501-6; Maclennan AH, Taylor AW, Wilson, Wilson D. The prevalence of pelvic floor disorders and their relationship to gender, age, parity and mode of delivery. Br J Obstet Gynaecol 2000;107:1460 70]. Pathophysiological mechanisms that have been proposed include pelvic floor denervation, direct trauma to the pelvic floor musculature, abnormal synthesis and degradation of collagen and defects in endopelvic fascia [Al-Rawi ZS, Al-Rawi ZT. Joint hypermobility in women with genital prolapse. Lancet 1982;I:439-41; Gilpin SA, Gosling JA. Smith ARB, Warrell DW. The pathogenesis of genitourinary prolapse and stress incontinence in women. A histological and histochemical study. Br J Obstet Gynaecol 1989;96:15-23; Smith ARB, Hosker GL, Warrell DW. The role of partial denervation of the pelvic floor in the aetiology of genitourinary prolapse and stress incontinence of urine. A neurophysiological study. Br J Obstet Gynaecol 1989;96:24-8; Allen RE, Hosker GL, Smith ARB, Warrell DW. Pelvic floor damage and childbirth: a neurophysiological study. Br J Obstet Gynaecol 1990;97:770-9]. The procedure of choice for reconstructive surgery to the vagina should be tailored to the individual patient and be of low morbidity and mortality, but at the same time with long-term durability. PMID- 17462359 TI - Publishing Surgeons Outcomes in the lay press is a bad idea - peer review of morbidity and mortality data is the way forward. PMID- 17462360 TI - Publishing Surgeons Outcomes in the lay press - call for a national debate. PMID- 17462361 TI - Oral sildenafil ameliorates impaired pulmonary circulation early after bidirectional cavopulmonary shunt. AB - We report a case of successful administration of oral sildenafil (ie, a phosphodiesterase-5 inhibitor) in an infant for impaired pulmonary circulation that caused early clinical deterioration after a bicavopulmonary shunt. The transpulmonary pressure gradient (ie, a clinical indicator of pulmonary circulation) was initially normalized by inhaled nitric oxide; however, an increase in transpulmonary pressure gradient and oxygen desaturation occurred after extubation and discontinuation of inhaled nitric oxide on postoperative day 1. Subsequent administration of oral sildenafil in stepwise doses resulted in normalization of transpulmonary pressure gradient and improved oxygen saturation with successful discontinuation of intravenous vasodilators. Our results suggest that oral sildenafil may be a potent adjunctive therapy for impaired postoperative pulmonary circulation after right heart bypass surgery. PMID- 17462362 TI - Effectiveness of hyperbaric oxygen therapy for hearing loss after cardiac surgery. AB - Sudden sensorineural hearing loss is a rare complication after cardiac surgery with extracorporeal circulation. We report a case of a 60-year-old man who experienced severe sensorineural hearing loss after Bentall procedure and recovery of normal hearing after 20 days of hyperbaric oxygen therapy. PMID- 17462363 TI - Actual and actuarial probabilities of competing risks: apples and lemons. AB - The probability of a type of failure that is not inevitable, but can be precluded by other events such as death, is given by the cumulative incidence function. In cardiac research articles, it has become known as the actual probability, in contrast to the actuarial methods of estimation, usually implemented by the Kaplan-Meier (KM) estimate. Unlike cumulative incidence, KM attempts to predict what the latent failure probability would be if death were eliminated. To do this, the KM method assumes that the risk of dying and the risk of failure are independent. But this assumption is not true for many cardiac applications in which the risks of failure and death are negatively correlated (ie, patients with a higher risk of dying have a lower risk of failure, and patients with a lower risk of death have a higher risk of failure, which is a condition called informative censoring). Recent editorials in two cardiac journals have promoted the use of the KM method (actuarial estimate) for competing risk events (specifically for heart valve performance) and criticized the use of the cumulative incidence (actual) estimates. This report has two aims: to explain the difference between these two estimates and to show why the KM is generally not appropriate. In the process we will rely on alternative representations of the KM estimator (using redistribution to the right and inverse probability weighting) to explain the difference between the two estimates and to show how it may be possible to adjust KM to overcome the informative censoring. PMID- 17462364 TI - Thoracic Surgery Directors Association Award. What is the optimal management of late-presenting survivors of acute type A aortic dissection? AB - BACKGROUND: Although type A aortic dissections represent a surgical emergency, some patients present late after the onset of symptoms. Optimal management of this cohort has not been defined. METHODS: Data on 195 patients with type A dissections followed up at a single institution between 1985 and 2005 were collected prospectively. Of these, 93 patients (47.2%) presented 48 hours or later after the initial onset of pain (group A), and the remaining 102 patients underwent immediate operative repair (group B). Median follow-up was 41.8 months (range, 0 to 386 months). RESULTS: Patients in group A were older (68.8 versus 59.3 years, p = 0.0005) and had a higher incidence of coronary artery disease (42.5% versus 14.6%, p < 0.0001), pulmonary disease (26.6% versus 8.4%, p = 0.0023), and congestive heart failure (14.1% versus 1.0%, p = 0.0004). Long-term survival was similar, although group B showed a trend toward improved 30-day mortality (16.5% versus 8.7%, p = 0.1035). Of the 92 patients in group A, 53 (57.6%) eventually underwent operative repair a median of 8.2 days after symptom onset. There was a trend toward improved long-term survival among patients undergoing repair (p = 0.1031). CONCLUSIONS: Initial medical management with interval operative repair of selected patients referred greater than 2 days following an acute type A dissection is a viable option. Delayed repair after optimization of the clinical condition and detailed evaluation of concomitant diseases results in excellent long-term results. PMID- 17462365 TI - Proximal reoperations after repaired acute type A aortic dissection. AB - BACKGROUND: Concerned with the associated risks of proximal reoperation, some have proposed an aggressive approach of aortic root replacement during emergent repair of acute type A aortic dissection. Because few data exist regarding late reoperations, we report outcomes of proximal reoperation after repaired type A aortic dissection. METHODS: Between January 1991 and March 2006, 63 patients underwent reoperation after previous repair for acute type A aortic dissection. Procedures performed at reoperation included ascending (94%, 59 of 63), total arch (62%, 39 of 63), elephant trunk (56%, 35 of 63), aortic valve replacement (38%, 24 of 63), aortic root (27%, 17 of 63), and coronary artery bypass graft (8%, 5 of 63). Preoperative, operative, and postoperative variables were analyzed retrospectively with regard to early and late mortality. RESULTS: Thirty-day mortality was 11.1% (7 of 63). No strokes occurred. Incidence of renal failure, respiratory failure, and bleeding was 6% (4 of 63), 23% (15 of 63), and 6% (4 of 63), respectively. Mean time from initial repair to reoperation was 69 months (range, 1 to 258). Procedure performed (root versus ascending/resuspension) at initial repair did not affect the time to reoperation (p > 0.05). Median follow up was 40 months; and 1-, 5-, and 10-year survival was 82%, 74%, and 62%, respectively. Multivariate predictors of late mortality were prior coronary artery bypass graft surgery (odds ratio = 6.5, p < 0.003), bypass time (odds ratio = 3.6, p < 0.02), and renal dysfunction (odds ratio = 3.7, p < 0.05). CONCLUSIONS: Proximal reoperations for repaired acute type A aortic dissection can be performed with acceptable early and late mortality. The concern for proximal reoperation should not dictate the initial procedure choice during acute type A aortic dissection. Continued clinical and radiographic surveillance of repaired type A aortic dissection is warranted. PMID- 17462366 TI - Reoperation for giant false aneurysm of the thoracic aorta: how to reenter the chest? AB - BACKGROUND: Giant false or pseudoaneurysm of the aorta is a rare but dreadful complication occurring several months or years after cardiac or aortic surgery. We describe a surgical approach that allowed safe reentry in the chest in five patients, with a mean follow-up of almost seven years. METHODS: From December 1991 to October 1999, five patients aged 34 to 74 years (mean age, 55 +/- 11.6 years), who had previously undergone a total of nine operations in other institutions, required reoperation for giant false aneurysm of the ascending aorta in a mean delay of 22.6 +/- 20.3 months (3 months to 6 years) after the last surgical procedure. In order to avoid major mediastinal wound and patient's exsanguination during resternotomy, the following technique has been used: femoral artery cannulation; right atrial cannulation through the femoral vein; femoro-femoral full-flow cardiopulmonary bypass; rectal temperature lowered to 25 degrees C; direct cannulation and cross-clamping of both carotid arteries through a direct cervical approach, and selective cerebral perfusion with cold blood (10 degrees C to 12 degrees C); circulatory arrest of the main circuit; chest opening; and mediastinal division. RESULTS: Despite the fact that the false aneurysm was entered in all patients, reopening of the chest has been safe in all cases. In four cases, the aortic repair consisted of complete graft replacement (Dacron) of the compromised aortic segment (ascending aorta in two; both ascending aorta and aortic arch in two). In one case, reimplantation of the left coronary ostium and closure of a fistula with the left ventricle was carried out. One patient with ongoing mediastinitis died from intractable septicemia and multiorgan failure. Presently, two patients are in excellent condition; one suffers from light neurologic sequelae (oculomotor nerves palsy) and one patient had a nonrelated stroke one year postoperatively. CONCLUSIONS: The technique of separate carotid cannulation and selective antegrade brain perfusion with cold blood during circulatory arrest at moderate core hypothermia has, in our opinion, many advantages. In addition to allowing harmless opening of the chest in the presence of most dangerous mediastinal false aneurysms, it implies no general deep hypothermia, reduced duration of cardiopulmonary bypass, and circulatory arrest of the lower part of the body, and safe and permanent brain protection throughout chest opening and mediastinal division. It has allowed us to safely reoperate on patients who are generally considered as a major surgical risk. PMID- 17462367 TI - Midterm results of aortic repair using a fabric neomedia and fibrin glue for type A acute aortic dissection. AB - BACKGROUND: Controversy exists concerning the optimal surgical treatment of acute type A aortic dissection to reduce mortality rate and the need for reoperation. The goal of the present study was to evaluate midterm results of repair using a fabric and fibrin glue for acute type A aortic dissection. METHODS: From 1994 to 2005, 100 patients with acute type A aortic dissection underwent supracommissural graft replacement using a fabric as "neomedia" and fibrin glue. Mean patient age was 65.7 +/- 11.3 years. RESULTS: Hospital mortality was 9%. All 91 survivors underwent follow-up evaluation for a mean period of 47.7 months. Aortic regurgitation was restored to non or mild in 46 survivors who had preoperative aortic regurgitation in the immediate period, and only one patient developed moderate aortic regurgitation in the midterm period. Survival at 1, 5, and 10 years including hospital mortality was 89.0 +/- 3.1%, 75.2 +/- 5.1%, and 59.2 +/- 10.9%, respectively. Reoperation for aortic lesion was performed without mortality in three patients (residual dissection, n = 1; sinus of Valsalva dilatation, n = 1; acute redissection, n = 1). During two late reoperations, the fabric was observed to be firmly adherent to the dissected wall and maintained the shape of the aorta. Freedom from aortic reoperation was 98 +/- 2% and 98 +/- 2%, at 5 and 10 years, respectively. CONCLUSIONS: Supracommissural aortic replacement using a fabric neomedia and fibrin glue resulted in low early and late mortality as well as a low reoperation rate. PMID- 17462368 TI - Late complications of gelatin-resorcin-formalin glue in the repair of acute type A aortic dissection. AB - BACKGROUND: During surgical treatment for acute type A aortic dissection, gelatin resorcin-formalin glue is generally applied and its efficacy has been reported. However, some late complications that are potentially associated with this glue have also been reported. In the present study, we reviewed our experiences of treatment for acute type A aortic dissection and late complications that occurred in the anastomotic site, which needed a reoperation. METHODS: From October 1994 to August 2005, 68 patients underwent emergency surgery for acute type A aortic dissection. Gelatin-resorcin-formalin glue was applied to 56 (82.4%) of these patients at one or both of the distal and proximal anastomosis sites. RESULTS: Eight (11.8%) patients died in hospital within 30 days after the operation, among which two patients already had cerebral complications prior to the surgery. There were five late deaths from causes unrelated to cardiac events. Five patients developed an aortic pseudoaneurysm at the anastomotic site and underwent a late reoperation. All of these patients had been treated with gelatin-resorcin formalin glue during the previous operation. Histologic examination of the resected aortic wall after the reoperation revealed tissue necrosis, severe local inflammation, and organization of old thrombi at the site of the glue application. CONCLUSIONS: Late complications after the use of gelatin-resorcin formalin glue may occur with a certain amount of risk, suggesting its toxicity for aortic tissue. Therefore, proper use of this glue and close follow-up of the patients are strictly required. PMID- 17462369 TI - Invited commentary. PMID- 17462370 TI - Survival benefit of endovascular descending thoracic aortic repair for the high risk patient. AB - BACKGROUND: Despite acceptable results reported with endovascular thoracic aortic repair (TEVAR), recent studies have questioned the merit of repair in asymptomatic patients considered high risk for open surgery. In this group, advanced age or comorbid conditions may reduce life expectancy, thus limiting the benefit of elective aneurysmectomy. This study was conducted to determine whether elective TEVAR improves survival for this cohort. METHODS: Forty-six asymptomatic patients with descending thoracic aortic disease were considered high risk for open surgery for reasons of age of 80 years or older (47.8%) or comorbid conditions (84.8%), and were subsequently evaluated for elective TEVAR. Of these, 21 underwent TEVAR, while another 25 patients were excluded from TEVAR on the basis of unfavorable anatomy or refused intervention. RESULTS: The mean age of the cohort was 77.0 +/- 7.0 years (p = 0.9 between groups). Prevalent comorbid conditions were similar between groups, and included coronary artery disease (p = 1.0), chronic obstructive pulmonary disease (p = 1.0), and peripheral vascular disease (p = 0.23). Mean maximum aortic diameter was 6.0 +/- 1.4 cm (p = 0.54 between groups). Indications for intervention included fusiform aneurysm (65.2%) and pseudoaneurysm or penetrating ulcer (32.6%). No 30-day mortality was observed after TEVAR. All-cause mortality in the entire cohort was 50%. Median actual time to mortality was different between groups (control, 9.2 months versus TEVAR, 24.9 months; p = 0.01). Life-table analysis demonstrated improved survival for TEVAR at 24 months (p = 0.05). CONCLUSIONS: Although the overall prognosis for the asymptomatic patient with descending thoracic aortic disease at high risk for open surgery is poor, elective endovascular repair improves survival and should be considered a therapeutic option in this setting. PMID- 17462371 TI - Endovascular stent-graft placement of aneurysms involving the descending aorta originating from chronic type B dissections. AB - BACKGROUND: The performance of endovascular stent-graft placement in patients suffering from aneurysms involving the descending aorta originating from chronic type B dissections is unclear. METHODS: Within a 2-year period, we treated 6 patients with this pathology. Four patients required extension of the proximal landing zone (autologous double transposition, n = 2; subclavian-to-carotid artery transposition, n = 2) before stent-graft placement. RESULTS: Supra-aortic rerouting procedures and endovascular stent-graft placement were performed successfully in all patients. Closure of the primary entry tear, full expansion of the stent-graft, and eventually, thrombosis of the false lumen was achieved in 5 patients. In 1 patient with a short proximal landing zone, a persisting type Ia endoleak was observed. In all patients with successful primary entry closure, a reduction in aneurysm diameter occurred. Mean follow-up is 16 months (range, 4 to 25). CONCLUSIONS: Endovascular stent-graft placement of aneurysms involving the descending aorta originating from chronic type B dissections may serve as a valuable treatment option in this complex pathology. The chronic dissection membrane can be successfully compressed against large areas of the native aortic wall. A sufficient proximal landing zone is mandatory for early and late success. PMID- 17462372 TI - Invited commentary. PMID- 17462373 TI - Optimized preservation of extracellular matrix in cardiac tissues: implications for long-term graft durability. AB - BACKGROUND: Cryopreservation of human tissues, particularly heart valves, is widespread in clinical practice although the effects of this process on underlying tissue structures and its potential impact on valve durability have been poorly studied. Multiphoton imaging and second-harmonic generation (SHG) microscopy permit high-resolution, noninvasive analysis of living tissues at a subcellular level. In the present study we used these novel imaging modalities to compare the effects of vitreous and frozen cryopreservation on the extracellular matrix (ECM) of cardiac tissues. METHODS: Conventional histology, electron microscopy, and multiphoton imaging to obtain autofluorescence and SHG images were performed on cardiac tissues to characterize the ECM in fresh, vitrified, and frozen cryopreserved tissues. RESULTS: Autofluorescence and particularly SHG images revealed that conventional frozen cryopreservation of cardiac valves, when compared with fresh or vitrified tissues, leads to the loss of normal ECM structures in valve leaflets. Similar results were found in all other cardiac tissues suggesting that structural deterioration of the ECM is a common consequence of frozen cryopreservation. CONCLUSIONS: Our results demonstrate that conventional cryopreservation, when compared with fresh or vitrified tissues, causes more destruction of normal ECM structure, which might contribute to eventual graft dysfunction. Whether vitrification preservation will translate into greater durability or less valve failure will need to be determined. PMID- 17462374 TI - Aortic valve replacement in octogenarians: risk factors for early and late mortality. AB - BACKGROUND: Excellent outcomes after aortic valve replacement (AVR) in elderly patients can be achieved, yet some practitioners are reticent to refer elderly patients for surgery. This study analyzed risk factors for mortality in patients aged 80 years and older undergoing AVR with or without concomitant coronary artery bypass grafting (CABG). METHODS: A retrospective review was performed of 245 patients (129 women) with a mean age of 83.6 +/- 2.9 years who had AVR with (n = 140) or without CABG (n = 105) at a single institution from 1993 to 2005. Data were analyzed with a multivariate logistic regression for predictors of operative mortality, Kaplan-Meier estimates of survival, and a Cox multivariate proportional analysis of factors influencing long-term survival. RESULTS: Mean preoperative New York Heart Association (NYHA) classification was 3.1 +/- 0.9, and 78% (192/245) of patients were classified as NYHA class III or IV. Operative (30-day) mortality was 9% (22/245). Independent risk factors for operative mortality included postoperative renal failure (odds ratio [OR], 20.9; 95% confidence interval [CI], 6.5 to 67.6; p < 0.001), postoperative permanent stroke (OR, 11.3; 95% CI, 1.7 to 75.1; p = 0.019), or intraoperative/postoperative intraaortic balloon pump (IABP) placement (OR, 14.9; 95% CI 2.9 to 75.8; p = 0.002). Survival after surgery was 82% (n = 183) at 1 year and 56% (n = 88) at 5 years. Prognostic factors for decreased long-term survival were regurgitant valve pathology (hazard ratio [HR], 6.0; 95% CI, 2.5 to 14.2; p = 0.002), intraoperative/postoperative IABP (HR, 2.9; 95% CI, 1.4 to 6.0; p = 0.010), postoperative renal failure (HR, 3.5, 95% CI, 2.2 to 5.7; p < 0.001), and postoperative stroke (HR, 7.0, 95% CI, 3.2 to 15.9; p < 0.001). Performing concomitant CABG was protective in terms of operative mortality (OR, 0.3; 95% CI, 0.09 to 0.83; p = 0.017) and improved long-term survival (HR, 0.7, 95% CI, 0.47 to 0.96; p = 0.020). Preoperative NYHA classification did not affect operative or long-term survival. CONCLUSIONS: Patients aged 80 years and older who undergo AVR have acceptable short-term and long-term survival regardless of NYHA status. Concomitant CABG improved operative and long-term survival in this population. Despite their increased age, aggressive surgical treatment is warranted for most patients. PMID- 17462375 TI - A simplified approach to degenerative disease: triangular resections of the mitral valve. AB - BACKGROUND: Only 40% of patients with mitral valve (MV) regurgitation undergo operative repair rather than replacement. Quadrangular resection combined with ring annuloplasty has been the most common method of repair for degenerative posterior leaflet disease. Techniques such as sliding annuloplasty and artificial chord usage have increased the complexity of repair. These techniques have been perceived to be difficult and have possibly reduced the incidence of MV repair. We present our experience with a simplified approach to MV repair utilizing a triangular resection and larger rings. METHODS: Retrospective review of all MV repairs over a 7-year period (1999 to 2006) revealed 154 patients who underwent triangular resection for degenerative disease. Patients who underwent ring annuloplasty without leaflet resection and patients who had artificial chords or quadrangular resections were excluded. RESULTS: Of 154 patients who underwent triangular resection, isolated posterior leaflet resection was performed on 130 patients. Isolated anterior and combined anterior and posterior leaflet triangular resections were performed on 16 and 8 patients, respectively. Thirty day postoperative mortality was 0%. Five-year freedom from reoperation for recurrent mitral regurgitation was 99.0%. No patients who had intended leaflet resection were converted to MV replacement. Intraoperative transesophageal echocardiogram revealed trace to 1+ mitral regurgitation. Mild systolic anterior motion was noted in 7.1% of cases initially, but resolved with volume loading in all. CONCLUSIONS: Triangular leaflet resection of the mitral valve produces durable results and can be safely and efficiently performed with minimal morbidity and mortality. This technique should allow increased utilization of MV repair for degenerative disease. PMID- 17462376 TI - Long-term results of bilateral internal thoracic artery grafting in dialysis patients. AB - BACKGROUND: We evaluated the perioperative and long-term results of bilateral internal thoracic artery grafting in dialysis patients. METHODS: One hundred one consecutive patients on chronic dialysis who underwent isolated coronary artery bypass grafting were retrospectively compared according to the surgical technique, bilateral internal thoracic artery (BITA) grafting (n = 76) or single internal thoracic artery (SITA) grafting (n = 25). RESULTS: Hospital mortality was 5.3% in the BITA group and 8.0% in the SITA group (p = not significant). The incidence of mediastinitis was not different (7.9% in the BITA group and 8.0% in the SITA group). The median duration of follow-up was 3.1 years (range, 0.1 to 10.9). Survival and freedom from cardiac mortality were not different between the two groups, but the BITA group had a trend toward better results. Freedom from cardiac events (including cardiac-related death, myocardial infarction, percutaneous coronary intervention, redo coronary artery bypass grafting, and congestive heart failure) was superior in the BITA group (p = 0.03). Calcification of the ascending aorta, peripheral vascular disease, insulin dependent diabetes mellitis, and age were the independent risk factors of late death. CONCLUSIONS: Perioperative results of BITA grafting in dialysis patients were not different from the results of SITA grafting. However, the long-term results of BITA grafting in dialysis patients were better than the results of SITA grafting. Overall, our results support the continued use of BITA grafting in dialysis patients. PMID- 17462377 TI - Bypass graft and native postanastomotic coronary artery patency: assessment with computed tomography. AB - BACKGROUND: Multidetector computed tomography has been shown to be useful in the evaluation of coronary artery bypass grafts in previous studies. We studied the accuracy of multidetector computed tomography in the detection of patency and significant stenosis of both grafts and native postanastomotic coronary arteries. METHODS: Ninety-six patients with 216 grafts (98 left mammary artery, 8 right mammary artery, 8 radial artery, and 102 venous grafts) were investigated by 16 slice computed tomography. Native postanastomotic coronary arteries were also evaluated. Patients unable to maintain a breath hold of 40 s were excluded. Computed tomography data were compared with the results of conventional angiography. RESULTS: On a segment-based model, the overall feasibility of computed tomography was 98.1% (212 of 216 grafts) for bypass grafts and 93.1% (201 of 216 segments) for postanastomotic coronary arteries. The leading cause of unfeasibility for postanastomotic coronary arteries was the small diameter of the examined vessel (<1.5 mm). Computed tomography correctly diagnosed all the 25 occluded grafts. Of the 33 significant stenoses of grafts, computed tomography correctly diagnosed 31. Sensitivity, specificity, positive predictive value, and negative predictive value were 100%, 98.5%, 96.5%, and 100%, respectively, for bypass graft; and 100%, 97.7%, 85%, and 100%, respectively, for coronary arteries. On a patient-based model, the feasibility, sensitivity, specificity, positive predictive value, and negative predictive value were 89.4% (86 of 96 patients), 100%, 93%, 86.4%, and 100%, respectively. CONCLUSIONS: Multidetector computed tomography allows a very accurate assessment of arterial and venous conduits and of postanastomotic native coronary arteries in patients with previous bypass graft. Despite high feasibility (93.1%), limitations of the method were breath-hold duration (35 to 40 s) and postanastomotic assessment of small vessels (which, however, precluded the analysis in only 4.6% of cases). PMID- 17462378 TI - Predictors of early postoperative cerebral infarction after isolated off-pump coronary artery bypass grafting. AB - BACKGROUND: Risk factors associated with cerebral infarction within 7 days after off-pump coronary artery bypass grafting require further statistical elucidation. METHODS: From January 1997 to July 2006, off-pump coronary artery bypass grafting was performed in 576 patients at Toyama University Hospital. Factors including previous cerebral infarction, lesions in head and neck vessels, preoperative cerebral ischemic symptoms, intraaortic balloon pump use, number of coronary lesions, number of coronary bypasses, sites of coronary bypasses, number of proximal anastomoses in the ascending aorta, use of aortic no-touch procedure, partial aortic clamping, automatic proximal anastomosis devices, and a proximal seal system were compared retrospectively between patients with and without early cerebral infarction. Factors differing between groups were analyzed further by multivariate logistic regression. RESULTS: Ten patients (1.7%) had cerebral infarction within 7 days after surgery. The subjects were divided into two groups, those who had cerebral infarction after surgery (n = 10) and those who did not (n = 566). Factors showing significant intergroup differences were a previous history of cerebral infarction (p < 0.001), preoperative presence of cerebral ischemic symptoms (p < 0.001), vascular lesions in head and neck vessels (p < 0.001), use of partial aortic clamping (p = 0.002), and postoperative atrial fibrillation within 7 days after surgery (p = 0.011). Multiple logistic regression analysis indicated close relationships between previous history of cerebral infarction (odds ratio, 26.6; 95% confidence interval, 2.8 to 251.1; p = 0.004), preoperative presence of cerebral ischemic symptoms plus lesions in head and neck vessels (odds ratio, 22.8; 95% confidence interval 1.8 to 285.7; p = 0.015), and use of partial aortic clamping (odds ratio, 11.1; 95% confidence interval, 1.4 to 85.7; p = 0.021). Postoperative atrial fibrillation within 7 days after surgery (odds ratio, 3.4; 95% confidence interval, 0.7 to 16.5; p = 0.121) was suspected as a risk factor for postoperative cerebral infarction. CONCLUSIONS: Multivariate analysis identified independent factors strongly associated with cerebral infarction after off-pump coronary artery bypass grafting, such as partial aortic clamping, presence of cerebral ischemic symptoms plus head and neck vascular lesions, and previous cerebral infarction. PMID- 17462379 TI - Significance of neurologic complications in the modern era of cardiac transplantation. AB - BACKGROUND: Severe neurologic complications after cardiac transplantation are devastating outcomes of this life-saving procedure. Incidence, risk factors, and morbidity of neurologic events in the modern era of cardiac transplantation are yet to be defined. METHODS: Between 1996 and 2005, 200 patients (64% men; mean age, 49 +/- 12 years) underwent heart transplantation at our institution. Overall, 46 patients (23%) showed a wide spectra of early neurologic complications. RESULTS: Cause of ischemic complications was stroke in 11 patients (7 had concomitant epileptic seizures) and transient ischemic attack (TIA) in 7. Encephalopathy (n = 10), epileptic seizures unrelated to focal cerebral lesions (n = 7), severe headache (n = 6), cerebral infection (n = 3), and peripheral nervous system injuries (n = 2) completed the spectra of adverse neurologic outcomes. Multivariate analysis identified advanced age (p = 0.03), preoperative left ventricular assist device support (p = 0.02), preoperative intraaortic balloon pump support (p < 0.001), prolonged cardiopulmonary bypass time (p < 0.001), and postoperative hepatic failure (p = 0.04) as independent predictors of early neurologic complications. Postoperative morbidities associated with neurologic complications included longer ventilation time (p < 0.001), longer stay in the intensive care unit (p < 0.001), and higher incidence of pneumonia (p < 0.001) and sepsis (p = 0.01) compared with patients without neurologic events. There was a trend toward higher in-hospital mortality (15% versus 6%, p = 0.07), but there was no difference in long-term survival (65% versus 78%, p = 0.15). CONCLUSIONS: Despite rigorous pretransplantation screening, neurologic complications were common after cardiac transplantation. Most frequently, these complications were not the actual cause of death, but they significantly contributed to increased morbidity. Preoperative mechanical circulatory support requirement was the strongest predictor for adverse neurologic outcome. PMID- 17462380 TI - Invited commentary. PMID- 17462381 TI - Orthotopic heart transplantation in patients with Marfan syndrome. AB - BACKGROUND: Due to the risk of vascular complications, the indication for heart transplantation (HTx) in patients with Marfan syndrome and end-stage heart disease remains controversial. We analyzed the results of such patients who underwent HTx at our institution. METHODS: Ten patients with Marfan syndrome (median age 36, range 19 to 56 years) underwent HTx between March 1986 and December 2005. The primary vascular manifestation of Marfan syndrome was type-A aortic dissection in three patients and ascending aortic aneurysm in seven patients. All patients had undergone cardiovascular operations prior to transplantation. All had refractory heart failure (New York Heart Association class IV) before transplantation. Three patients underwent transplantation after ventricular assist device (VAD) support (left VAD, n = 2; biventricular assist device, n = 1). RESULTS: There were no perioperative deaths. Two patients died of causes unrelated to Marfan disease (pneumonia on day 27, n = 1; stroke on day 102, n = 1). One patient died due to type-B dissection 3.8 years posttransplantation and one due to rupture of an aortic arch aneurysm after 12.1 years. Two patients underwent thoracoabdominal aortic replacement for chronic dissection 14 and 20 months posttransplantation, respectively. Kaplan-Meier survival rate was 80% at 1 year and 64% at 10 years. The Kaplan-Meier freedom from reoperation was 100% at 1 year and 62.5% at 10 years. CONCLUSIONS: Heart transplantation in patients with Marfan syndrome results in good long-term survival, similar to that of patients without Marfan syndrome. Close follow-up and timely operation of aortic pathologies is mandatory. Reluctance to place these patients on a heart transplant waiting list appears not to be justified. PMID- 17462382 TI - Invited commentary. PMID- 17462383 TI - Medically refractory pulmonary hypertension: treatment with nonpulsatile left ventricular assist devices. AB - BACKGROUND: Severe pulmonary hypertension refractory to medical treatment is a contraindication to orthotopic heart transplantation in most centers. We report our experience in treating severe pulmonary hypertension with mechanical left ventricular unloading using implantable nonpulsatile left ventricular assist devices (LVAD) with continuous flow properties. METHODS: In ten patients with severe pulmonary hypertension, refractory to medical treatment, an implantable nonpulsatile LVAD was placed for continuous mechanical left ventricular support. Pulmonary hemodynamics were assessed by right heart catheterization prior to and during LVAD implantation, and after orthotopic heart transplantation. RESULTS: The mean (+/-SD) interval of nonpulsatile support was 182 (+/-118) days. Pulmonary artery pressure (mean +/- SD) significantly decreased from 42 +/- 13 to 24 +/- 5 mm Hg (p < 0.005), the transpulmonary gradient (mean +/- SD) decreased from 20 +/- 6 to 11 +/-5 mm Hg (p < 0.005), and the pulmonary vascular resistance (mean +/- SD) from 4.8 +/- 1.8 to 2.2 +/- 0.8 Wood units (p < 0.005) during an interval of one to six months of LVAD support. No significant increases in pulmonary artery pressure, transpulmonary gradient, and pulmonary vascular resistance were observed during an interval of three to six months after orthotopic heart transplantation. CONCLUSIONS: This study supports that LVAD support and continuous nonpulsatile mechanical unloading of the left ventricle can reverse medically unresponsive pulmonary hypertension and render patients eligible for orthotopic heart transplantation. PMID- 17462384 TI - Invited commentary. PMID- 17462385 TI - Atrial fibrillation after coronary artery bypass grafting surgery: development of a predictive risk algorithm. AB - BACKGROUND: Atrial fibrillation is a costly complication occurring in 15% to 40% of patients after coronary artery bypass grafting (CABG). Aggressive prophylactic treatment should be directed toward and limited to selected high-risk patients. Utilizing perioperative risk factors, we sought to develop an algorithm to predict the relative risk of developing postoperative atrial fibrillation in patients undergoing CABG. METHODS: Data were extracted from our Society of Thoracic Surgeons Database on 19,620 patients undergoing CABG between January 1995 and July 2006. We used perioperative risk factors to develop a logistic regression equation predictive for the development of postoperative atrial fibrillation. A total of 19,083 patients had complete data and were used to construct the final model. The model was used to compare the predicted probability of atrial fibrillation with the known outcome in the patients divided into deciles by probability. Bootstrap procedures were used to determine the confidence limits of the beta coefficients. RESULTS: A regression model was developed with 14 significant indicators. Those showing the greatest predictive influence included the patient age, the need for prolonged ventilation (24 hours or more), the use of cardiopulmonary bypass, and preoperative arrhythmias. The model showed acceptable concordance between observed and predicted (72.3%), a receiver operating characteristic curve area of 0.72, and Hosmer-Lemeshow probability of 0.19. When applied to the patient population, the calculated risk in those who did not develop AF was 0.179 +/- 0.116 and for those with AF, 0.284 +/- 0.153 (p < 0.001). CONCLUSIONS: A validated predictive risk algorithm for developing postoperative atrial fibrillation can reliably stratify patients undergoing CABG into high-risk and low-risk groups. This may be used preoperatively to appropriately target high-risk patients for aggressive prophylactic treatment. PMID- 17462386 TI - Health care consumption due to atrial fibrillation is markedly reduced by Maze III surgery. AB - BACKGROUND: Health care consumption and costs for the treatment of atrial fibrillation are high. Atrial fibrillation is effectively treated by the surgical Maze III procedure according to the Cox method. We describe the effects of this procedure on health care consumption and economy. METHODS: From October 1997 through March 2002, 72 patients underwent the Maze III procedure. Medical records of these patients were reviewed, and all data regarding hospitalization and outpatient clinic visits for atrial fibrillation and its related diseases were recorded. Accounting divisions from the contributing hospitals were consulted for the exact cost of each of these services, which were allocated into preoperative, perioperative, and postoperative periods. RESULTS: The perioperative mortality was zero. Long-term freedom from symptomatic atrial fibrillation was verified in 96% of the patients. The number of hospitalization days decreased by 84%, from 471 during the preoperative period to 79 in the postoperative (p < 0.001), and costs during the same periods decreased by 75%, from 7,075,000 Swedish Kronor to 1,757,000 Swedish Kronor (p < 0.001). CONCLUSIONS: The Maze III procedure significantly decreased the postoperative hospitalization costs in patients undergoing surgery primarily for atrial fibrillation. As well as providing an effective treatment for symptomatic arrhythmia, this procedure breaks the undesirable trend of increasing health care consumption resulting from treatment of atrial fibrillation. PMID- 17462387 TI - Prevention of atrial flutter with cryoablation may be proarrhythmogenic. AB - BACKGROUND: Atrial flutter is a serious problem after surgery for congenital heart disease. METHODS: We performed an intraoperative linear one-minute cryolesion between a right atriotomy and the tricuspid annulus to prevent atrial flutter in 17 consecutive adult patients undergoing surgery for congenital heart disease. Coronary angiography and electrophysiology study using an electroanatomic mapping system to assess the conduction across the line and to try to induce atrial flutter were performed three months after the operation in 15 patients. RESULTS: Eleven patients had bidirectional block in the cryolesion, four patients did not, and two patients refused the electrophysiology study and coronary angiography. All patients with terminal temperature below -151 degrees C had bidirectional block, while only one patient with terminal temperature above 151 degrees C had bidirectional block. No patient with bidirectional block and all patients without bidirectional block had inducible or spontaneous atrial flutter (p = 0.0007). No lesion of the right coronary artery was detected at coronary angiography. CONCLUSIONS: The success rate was suboptimal and the intervention is potentially proarrhythmogenic in patients without block. Preventive strategies targeting atrial flutter should be validated with regard to the block rate achieved. PMID- 17462388 TI - Invited commentary. PMID- 17462389 TI - Video-assisted bilateral epicardial pulmonary vein isolation for the treatment of lone atrial fibrillation. AB - BACKGROUND: This paper aims to evaluate the feasibility and the efficacy of a new off-pump, bilateral thoracoscopic pulmonary vein isolation technique in patients with lone atrial fibrillation. METHODS: Between April 2004 and February 2006, 26 drug-resistant and symptomatic lone atrial fibrillation patients (18 permanent, 8 paroxysmal) underwent an irrigated radiofrequency ablation procedure using the Cardioblate ablation system (Medtronic, Minnesota). There were 16 men and 10 women with a mean age of 55 +/- 11 years. Mean duration of atrial fibrillation was 34.2 +/- 18.9 months. All patients underwent a bilateral thoracoscopic procedure in which both pulmonary veins were ablated with an atrial cuff using an off-pump epicardial approach. The conduction block was assessed by pacing the pulmonary veins after each ablation. Sixteen patients underwent endoscopic stapling of the left atrial appendage. RESULTS: There were no hospital deaths. All procedures were completed as planned without any conversions to sternotomy. There were no major complications. Follow-up was complete at 6 months, and 80% of the patients were in sinus rhythm (paroxysmal: 100%, permanent: 72%). Of the patients with permanent atrial fibrillation, 85% had regained their atrial transport function. No major thromboembolic event was observed during the follow up period. CONCLUSIONS: The video-assisted bilateral pulmonary vein isolation technique was safe and effective. It was curative for paroxysmal atrial fibrillation patients and effective for permanent atrial fibrillation cases. This technique may find wider application if accumulating data further support these findings. PMID- 17462390 TI - Novel techniques for tumor thrombectomy for renal cell carcinoma with intraatrial tumor thrombus. AB - BACKGROUND: Radical nephrectomy with tumor thrombectomy in patients with renal cell carcinoma and level I to III thrombus extension is directly associated with an improved prognosis. However, radical surgery in patients with level IV thrombus extension is associated with high perioperative mortality, even if long term survival is possible. In this report, we describe an alternative technique of vena caval and intraatrial tumor thrombectomy to decrease perioperative mortality and morbidity. METHODS: A cohort of 6 patients aged 46, 50, 53, 56, 54, and 52 years underwent radical nephrectomy with tumor thrombectomy from the vena cava and right atrium under mild hypothermic cardiopulmonary bypass and intermittent cross-clamping of the supraceliac abdominal aorta. Intraatrial tumor thrombectomy was performed on a beating, perfused heart in 4 patients and a hypothermic, cardioplegia-perfused heart in 2 patients. RESULTS: There were no early or late deaths. The aortic cross-clamp time was 12 and 15 minutes for patients 5 and 6, respectively. The cumulative hepatic and renal ischemic time was 16 minutes (range, 14 to 22 minutes) at 32 degrees C. The mean cardiopulmonary bypass time was 53.3 +/- 8.9 minutes (range, 40 to 65 minutes). At a mean follow-up of 43 +/- 24.6 months (range, 10 to 70 months), all patients are active and remain disease-free. CONCLUSIONS: We conclude that radical nephrectomy and tumor thrombectomy in patients with level IV thrombi can be safely performed with cardiopulmonary bypass, mild hypothermia. and intermittent supraceliac abdominal aortic occlusion, avoiding potential hematologic, hepatic, renal, neurologic, and septic complications associated with circulatory arrest. PMID- 17462391 TI - Positive heparin-platelet factor 4 antibody complex and cardiac surgical outcomes. AB - BACKGROUND: Given the large number of patients undergoing cardiac operations annually, it is important to identify populations at high risk for adverse outcomes. This observational study was conducted to determine the incidence of preoperative heparin-platelet factor 4 (HPF4) antibodies and to assess the associated risk of postoperative adverse outcomes in a nonselected cardiac surgery patient population. METHODS: Between March 2002 and December 2004, 1114 (92%) of 1209 patients undergoing cardiac surgery with heparin were tested in an unselected manner for HPF4 antibodies. Main outcome measures were HPF4 antibody seropositivity and fatal and nonfatal adverse clinical outcomes after cardiac surgery. RESULTS: Of those screened, 60 (5.4%) of 1114 had positive HPF4 antibodies preoperatively. These patients had longer mean postoperative length of stay (14.0 days versus 9.8 days, p = 0.05), a higher incidence of prolonged (> or = 96 hours) mechanical ventilation (20.3% versus 9.2%, p = 0.02), acute limb ischemia (5.1% versus 0.9%, p = 0.03), renal complications including dialysis (20.3% versus 10.5%, p = 0.03), and gastrointestinal complications (15.3% versus 5.9%, p = 0.01). Stepwise logistic regression analysis showed positive HPF4 antibody status to be an independent predictor for adverse outcome and was associated with a higher risk for renal complications, including dialysis (adjusted odds ratio 2.2; 95% confidence interval, 1.1 to 4.3), than was diabetes. CONCLUSIONS: In this large patient series, the presence of HPF4 antibodies before surgical heparin administration was an independent and clinically significant risk factor for postoperative adverse events after cardiac surgery. An optimal preoperative cardiac surgery risk profile should include HPF4 antibody status. PMID- 17462392 TI - Prognostic significance of elevated cardiac troponin I after heart surgery. AB - BACKGROUND: Cardiac troponin I (cTnI) measured after heart surgery has been associated with operative mortality. We sought to determine whether measuring cTnI after heart surgery provides additional prognostic information beyond that provided by validated preoperative risk scores, the Veterans Affairs (VA) risk score and the European System for Cardiac Operative Risk Evaluation (EuroSCORE). METHODS: We retrospectively collected cTnI levels measured 24 hours after surgery in 1,186 patients who underwent coronary artery bypass graft surgery (n = 696) or valve surgery (n = 490). The outcomes were operative death and perioperative myocardial infarction. The ability of the cTnI and the risk scores to discriminate patients who did or did not have the study outcomes was assessed by the area under the receiver operating curve (c-index). RESULTS: Mean age was 66 +/- 10 years. Median cTnI was 38 ng/mL after valve surgery versus 18 ng/mL after coronary artery bypass graft surgery (p < 0.0001). There were 51 operative deaths (4.3%) and 142 perioperative myocardial infarctions (12%). For every 50 ng/mL increase in cTnI, the odds of operative death increased by 40% (odds ratio, 1.4; 95% confidence interval: 1.2 to 1.6) after coronary artery bypass graft surgery and by 30% (odds ratio, 1.3; 95% confidence interval: 1.1 to 1.5) after valve surgery. Cardiac troponin I was a significant independent correlate of perioperative myocardial infarction and death (p < 0.0001) with a c-index of 0.70 for death. Addition of cTnI improved the c-indexes of the risk scores for predicting death (from 0.75 to 0.79 for the VA risk score; p = 0.1; and from 0.69 to 0.77 for the EuroSCORE; p = 0.005). CONCLUSIONS: Postoperative cTnI measured 24 hours after heart surgery is independently associated with operative death and perioperative myocardial infarction and improves the ability to predict operative mortality in comparison with preoperative risk scores alone. PMID- 17462393 TI - Does normothermic normokalemic simultaneous antegrade/retrograde perfusion improve myocardial oxygenation and energy metabolism for hypertrophied hearts? AB - BACKGROUND: Beating-heart valve surgery appears to be a promising technique for protection of hypertrophied hearts. Normothermic normokalemic simultaneous antegrade/retrograde perfusion (NNSP) may improve myocardial perfusion. However, its effects on myocardial oxygenation and energy metabolism remain unclear. The present study was to determine whether NNSP improved myocardial oxygenation and energy metabolism of hypertrophied hearts relative to normothermic normokalemic antegrade perfusion (NNAP). METHODS: Twelve hypertrophied pig hearts underwent a protocol consisting of three 20-minute perfusion episodes (10 minutes NNAP and 10 minutes NNSP in a random order) with each conducted at a different blood flow in the left anterior descending coronary artery (LAD [100%, 50%, and 20% of its initial control]). Myocardial oxygenation was assessed using near-infrared spectroscopic imaging. Myocardial energy metabolism was monitored using localized phosphorus-31 magnetic resonance spectroscopy. RESULTS: With 100% LAD flow, both NNAP and NNSP maintained myocardial oxygenation, adenosine triphosphate, phosphocreatine, and inorganic phosphate at normal levels. When LAD flow was reduced to 50% of its control level, NNSP resulted in a small but significant decrease in myocardial oxygenation and phosphocreatine, whereas those measurements did not change significantly during NNAP. With LAD flow further reduced to 20% of its control level, both NNAP and NNSP caused a substantial decrease in myocardial oxygenation, adenosine triphosphate, and phosphocreatine with an increase in inorganic phosphate. However, the changes were significantly greater during NNSP than during NNAP. CONCLUSIONS: Normothermic normokalemic simultaneous antegrade/retrograde perfusion did not improve, but slightly impaired myocardial oxygenation and energy metabolism of beating hypertrophied hearts relative to NNAP. Therefore, NNSP for protection of beating hypertrophied hearts during valve surgery should be used with extra caution. PMID- 17462394 TI - Invited commentary. PMID- 17462395 TI - Attenuated renal and intestinal injury after use of a mini-cardiopulmonary bypass system. AB - BACKGROUND: Transient, subclinical myocardial, renal, intestinal, and hepatic tissue injury and impaired homeostasis is detectable even in low-risk patients undergoing conventional cardiopulmonary bypass (CPB). Small extracorporeal closed circuits with low priming volumes and optimized perfusion have been developed to reduce deleterious effects of CPB. METHODS: A prospective, randomized trial was conducted in 49 patients undergoing elective coronary artery bypass graft surgery either with the use of a standard or mini-CPB system (Synergy). We determined early postoperative inflammatory response (leukocytosis, C-reactive protein, urine interleukin-6), platelet consumption and activation (urine thromboxane B2), proximal renal tubular injury (urine N-acetyl-glucosaminidase), and intestinal injury (intestinal fatty acid binding protein). RESULTS: In patients undergoing coronary artery bypass grafting with a mini-CPB system, we observed decreased priming volumes with subsequent attenuation of on-pump hemodilution, improved hemostatic status with reduced platelet consumption and platelet activation, decreased postoperative bleeding and minimized transfusion requirements. We also found reduced leukocytosis and decreased urinary interleukin-6. Levels of urine N acetyl-glucosaminidase were on average threefold lower, and urinary intestinal fatty acid binding protein was 40% decreased in the patients on the mini-CPB system, as compared with standard CPB. CONCLUSIONS: The use of the mini-CPB system during myocardial revascularization represents a viable nonpharmacologic strategy that can attenuate the alterations in the hemostatic system, reduce bleeding and transfusion requirements, decrease systemic inflammatory response, and reduce immediate postoperative renal and intestinal tissue injury. PMID- 17462396 TI - Invited commentary. PMID- 17462397 TI - Outcomes after bidirectional Glenn operation: Blalock-Taussig shunt versus right ventricle-to-pulmonary artery conduit. AB - BACKGROUND: There are distinct physiologic differences between patients with single-ventricle lesions who have undergone the Norwood procedure with a right ventricle-to-pulmonary artery conduit (NW-RVPA) compared with those patients who have undergone the Norwood operation with a Blalock-Taussig shunt (NW-BTS). We evaluated bidirectional Glenn operation outcomes and compared the two groups to assess whether the type of Norwood operation influenced outcomes. METHODS: A retrospective chart review compared bidirectional Glenn operation outcomes for children undergoing the Norwood operation with NW-RVPA or NW-BTS at Children's Hospital Boston from January 1, 2002, to December 31, 2003. RESULTS: Of 80 patients undergoing the Norwood operation, 56 (NW-BTS, 27 versus NW-RVPA, 29) returned for the bidirectional Glenn operation at our institution. The NW-RVPA group had a lower median age at presentation for bidirectional Glenn (4.5 months versus 5.8 months; p = 0.01), but had better weight gain (20.6 g/day versus 16.5 g/day; p = 0.03) than the NW-BTS group. No interstage deaths occurred in the NW RVPA group. There were no differences in morbidity or mortality after the BDG between the two groups. CONCLUSIONS: There were no differences in morbidity and mortality outcomes after the bidirectional Glenn operation between the NW-RVPA and NW-BTS groups. Despite younger age at presentation, the NW-RVPA patients had better growth rate, which may have contributed to the similar postoperative outcomes. PMID- 17462398 TI - Geometric disproportion of cardiac structure and graft ischemia affect tricuspid valve regurgitation early after neonatal heart transplantation. AB - BACKGROUND: Although tricuspid valve regurgitation (TR) after heart transplantation is a known complication, there has been little discussion of this subject in neonatal heart transplantation. We aim to elucidate the prevalence, etiology, and evolution of TR early after transplant in neonates. METHODS: Eighty five neonatal recipients were studied retrospectively by two-dimensional and Doppler echocardiography. The semiquantitative grading of TR was based on the ratio of regurgitation jet area to right atrial area. RESULTS: Immediately after neonatal heart transplantation, TR was recognized in 47 patients (grade 1, n = 18; grade 2, n = 22; grade 3, n = 7; and grade 4, n = 0). Tricuspid regurgitation prevalence diminished from 55% to 19% with reduction in severity 1 year after transplantation. The prevalence of TR (grade 2 and grade 3) was affected by a donor/recipient body weight ratio of more than 2.0 (p = 0.004) and graft ischemia for more than 3 hours (p = 0.014). The ratio of donor and recipient right atria portion, which had a correlation with donor/recipient body weight ratio (r2 = 0.415, p < 0.0001), separated the four subgroups in terms of TR grade immediately after transplantation (p = 0.0064) and also at 1 year after transplantation in all surviving grafts from 1.48 +/- 0.54 to 0.8 +/- 0.32 (p < 0.0001). The Cox model found no significance for early posttransplant TR as a risk factor for graft survival. CONCLUSIONS: Early posttransplant TR was affected by atria geometrical disproportion and by graft ischemia. Tricuspid regurgitation was not a risk factor for graft survival because of its amelioration over time, perhaps induced by recipient growth and recovery of myocardial injury relating to graft procurement. PMID- 17462399 TI - Incidence of and risk factors for pulmonary autograft dilation after Ross aortic valve replacement. AB - BACKGROUND: The Ross procedure is an alternative to mechanical aortic valve replacement in the young. Early dilation of the pulmonary autograft root exposed to the systemic circulation has been reported. The aim of our study is to define the prevalence, risk factors, and consequences of autograft dilation. All consecutive adult and pediatric patients who underwent Ross procedure at our institution were retrospectively reviewed for autograft dilation. METHODS: Between 1993 and 2005, 170 patients (mean age, 24.9 +/- 15.5 years; range, 1 month to 61 years) underwent Ross aortic valve replacement: 48% were younger than 19 years old. Eighty-seven additional procedures were performed in 58 patients (34%) at the time of the Ross procedure. End points of the study were freedom from autograft dilation (z value more than +2.0), autograft dysfunction, autograft reoperation, and autograft replacement. RESULTS: There were 2 early and 1 late deaths during a mean follow-up of 5.1 +/- 3.0 years (range, 1 month to 12 years). Actuarial survival at 10 years was 98%. Autograft dilation was identified in 31 patients (19%). Regurgitation (>2+) was identified in 12 patients (7%); all 12 had autograft dilation. At 10 years, freedom from autograft dilation was 82%, freedom from autograft dysfunction was 92%, freedom from reoperation on autograft was 92%, and freedom from autograft replacement was 96%. Cox proportional hazard analysis identified preoperative aortic annulus dilation (z value more than +2.0; p = 0.004), younger age (p = 0.05), time of surgery (before 2001; p = 0.002), and male sex (p = 0.01) as predictive of autograft dilation, whereas preoperative ascending aorta diameter (p = 0.01), male sex (p = 0.03), and postoperative systemic hypertension (p = 0.05) were predictive of autograft dysfunction. CONCLUSIONS: Significant autograft dilation is not common after the Ross procedure. Significant autograft dysfunction affects a minority of patients, but it is more prevalent in those with autograft dilation. PMID- 17462400 TI - Long-term functional assessment after correction of tetralogy of Fallot in adulthood. AB - BACKGROUND: Tetralogy of Fallot presenting in adulthood is a surgical challenge. We present the long-term outcomes of surgical correction in this subset of patients, including results of postoperative effort tolerance as assessed by treadmill testing. METHODS: Fifty-eight patients older than 18 years operated on between January 1995 and June 2004 are included in the study. Mean age at surgery was 22.5 +/- 5 years. Forty-seven patients were in New York Heart Association functional class II and 11 were in class III. Two patients had previous shunts. Forty-four patients received a transannular patch, and 14 had a right ventricular outflow tract patch. The prospective arm objectively assessed postoperative ventricular function by treadmill testing and echocardiography. RESULTS: Hospital mortality was 6.9%. Follow-up was 89% complete, with mean follow-up of 69.9 +/- 43 months. Late mortality occurred in 2 patients, both with infective endocarditis. Significant improvement in functional class was demonstrated (p < 0.001). Eight patients had significant pulmonary regurgitation on follow-up. The probability of survival after repair was 89% at 15 years. Thirty-five of 36 patients who underwent treadmill testing had good effort tolerance, with an average of 10.47 +/- 1.4 metabolic equivalents achieved. None had a positive result. One patient with transannular patch, in functional class III, had fair exercise tolerance with severe pulmonary regurgitation on echocardiography. CONCLUSIONS: Repair of adult tetralogy of Fallot has acceptable morbidity and mortality rates with good long-term surgical outcome in terms of effort tolerance as demonstrated by treadmill testing. Transannular patching does not appear to be a significant risk factor for right ventricular failure at long-term follow-up. PMID- 17462401 TI - Invited commentary. PMID- 17462402 TI - Evaluation of the coagulation system in children with two-ventricle congenital heart disease. AB - BACKGROUND: Multiple coagulation factor abnormalities involving both procoagulant and anticoagulant proteins have been described in children with single-ventricle physiology. This study used age-matched controls to evaluate coagulation factors in children with two-ventricle congenital heart disease (CHD). METHODS: Coagulation factors were assayed in 120 patients with CHD, divided into four age groups: group 1, 0 to 3 months; group 2, 3 to 12 months; group 3, 12 to 48 months; and group 4, older than 48 months. Healthy children without CHD were assayed as controls. Concentration of factors II, V, VII, VIII, IX, and X; protein C and S, plasminogen, and antithrombin III, were measured by standard assays. Normal ranges were determined by the empirical 95% confidence intervals. RESULTS: Significant reductions were found in mean levels of both procoagulant and anticoagulant factors in patients in groups 1, 2, and 3 compared with controls, but no differences were found in group 4. In group 1, all variables had significantly lower concentrations except fibrinogen and protein S; in group 2, all variables had significantly lower concentrations except for fibrinogen, factors VIII and IX, and plasminogen and protein S; and in group 3, all variables had significantly lower concentrations except fibrinogen, factors VIII and IX, and antithrombin III, plasminogen, and protein S. CONCLUSIONS: Neonates and infants with two-ventricle CHD have lower levels of procoagulant and anticoagulant factors compared with aged-matched controls approaching normal levels in children aged older than 4 years. These coagulation factor abnormalities are similar to those described in patients with single-ventricle physiology. PMID- 17462403 TI - Invited commentary. PMID- 17462404 TI - Propensity-matched analysis of three techniques for intrathoracic esophagogastric anastomosis. AB - BACKGROUND: A cervical side-to-side stapled esophagogastric anastomosis appears to decrease morbidity compared with traditional hand-sewn techniques. We evaluated our experience with this novel technique in intrathoracic anastomoses and compared the outcome with circular-stapled or hand-sewn techniques. METHODS: All patients undergoing transthoracic esophagectomy from 1999 to 2005 for esophageal cancer with gastric replacement were reviewed. A prospective quality improvement database, telephone interview, and chart review were used to collect data. A side-to-side stapled anastomosis was done in 44 patients, circular stapled anastomosis in 147, and hand-sewn anastomosis in 23. Propensity scores were generated from 14 variables, which were then used to generate 23 patient triplets. End points included leak, dysphagia, stricture, other major complications, and overall survival. Follow-up was available on all patients. RESULTS: For matched triplet comparison, no significant difference was noted in anastomotic leaks (8.7% with side-to-side stapled, 4.3% with circular-stapled, and 4.3% with hand-sewn; p = 0.78). Postoperative dysphagia was significantly higher in hand-sewn anastomoses at 56.5% versus 26.1% with side-to-side stapled and 21.7% with circular-stapled (p = 0.04). Stricture requiring esophageal dilation was also increased in hand-sewn at 34.8% versus 8.7% with side-to-side stapled and 8.7% with circular-stapled (p = 0.04). No difference was noted in perioperative mortality, long-term survival, or locoregional recurrences between techniques. CONCLUSIONS: In this carefully matched group of patients, intrathoracic use of the side-to-side stapled esophagogastric anastomosis in esophageal cancer patients is safe and effective. Postoperative dysphagia and need for stricture dilation may be decreased using a stapled compared with a traditional hand-sewn anastomosis. PMID- 17462405 TI - Immunopathological patterns of the stomach in adenocarcinoma of the esophagus, cardia, and gastric antrum: gastric profiles in Siewert type I and II tumors. AB - BACKGROUND: The morphologic and immunohistochemical profiles of gastric mucosa and of the tumor were assessed in Siewert type I, type II, and gastric antrum adenocarcinomas. METHODS: Sixty-two patients, prospectively operated upon, were included in the study: 37 type II, 15 type I, and 10 antrum adenocarcinoma. Samples of the tumor, the surrounding area, and the gastric corpus and antrum were analyzed histologically, and immunostained for cytokeratins (CK)7/20 (staining positive for cells labeled > or = 50%). RESULTS: Among the 37 type II adenocarcinomas were the following: (1) 13 of 37 (35%) had intestinal metaplasia (IM) in the stomach; (2) 24 of 37 (65%) did not show IM at any level; (3) 34 of 37 (92%) had Helicobacter pylori (HP) infection; (4) 13 of 37(35%) had CK7/20 expression of "Barrett's type" (CK7+/20-); 24 of 37 (65%) had a "no Barrett's type" profile (10 of 37 with CK7-/CK20+ and 14 of 37 with CK7+/CK20+); (5) 100% showed the same CK immunoprofile, both in IM and adenocarcinoma (measure of agreement k = 1, p = 0.000). Type I adenocarcinomas showed the following: (1) 87.5% CK Barrett's type, both in the tumor, and in the surrounding IM; (2) 100% gastric samples devoid of both IM and HP infection. Comparison between CK immunoprofiles in type I and type II tumors showed a difference within the two groups (p = 0.002). One hundred percent of antrum adenocarcinomas showed a no Barrett's type CK profile, both in the tumor and in the IM of the entire stomach. CONCLUSIONS: Data suggest that type II adenocarcinoma cannot be always considered a gastroesophageal reflux disease-related tumor; other pathogenetic pathways should be taken into consideration. PMID- 17462406 TI - Stereotactic radiosurgery for the treatment of lung neoplasm: initial experience. AB - BACKGROUND: Surgical resection is the standard of care for patients with resectable non-small cell lung carcinoma (NSCLC) or limited pulmonary metastases. Stereotactic radiosurgery (SRS) may offer an alternative option for high-risk patients who are not surgical candidates. We report our initial experience with SRS in the treatment of lung neoplasm. METHODS: Patients who were medically inoperable were offered SRS. Thoracic surgeons evaluated all patients, placed fiducials, and planned treatment in collaboration with radiation oncologists. A median dose of 20 Gy prescribed to the 80% isodose line was administered in a single fraction. The initial response rate, time to progression, and overall survival were evaluated. RESULTS: During a 2-year period, 32 patients, 27 with NSCLC and 5 with pulmonary metastases, underwent SRS. Fiducial placement resulted in a pneumothorax requiring a pigtail catheter in 9 patients (28%). An initial complete response was observed in 7 patients (22%) and partial response in 10 (31%). Disease was stable in 9 (28%) and progression occurred in 5 patients (16%), with a median time to local progression of 11 months. The median overall survival was 26 months. The probability of 1-year overall survival was 78% (95% confidence interval [CI], 65% to 94%) for the entire group and 91% (95% CI, 75% to 100%) for stage I patients. CONCLUSIONS: Our preliminary experience indicates that SRS has reasonable results in this high-risk group of patients, with pneumothorax being a significant morbidity. Surgery continues to offer the best chance of cure for resectable patients; however, SRS offers an alternative to high-risk patients. PMID- 17462407 TI - Ratio of the maximum standardized uptake value on FDG-PET of the mediastinal (N2) lymph nodes to the primary tumor may be a universal predictor of nodal malignancy in patients with nonsmall-cell lung cancer. AB - BACKGROUND: The maximum standardized uptake value (maxSUV) on F-18 fluorodeoxyglucose-positron emission tomography (FDG-PET) scan of mediastinal (N2) lymph nodes may predict pathology in patients with nonsmall-cell lung cancer. However, the maxSUV varies among PET scanners. Thus, we evaluated the ratio of the maxSUV of the lymph node to the primary tumor at different centers to determine whether it was a universal predictor of lymph node malignancy. METHODS: This is a retrospective review of a prospective database. Patients with nonsmall-cell lung cancer, a dedicated FDG-PET with the maxSUV of the primary lung tumor and FDG-avid mediastinal (N2) nodes reported (before therapy), and who underwent lymph node removal were eligible. RESULTS: There were 239 patients with 335 FDG-PET-positive N2 nodes at 14 different PET centers. The median ratio of the maxSUV of the lymph node to the maxSUV of the primary tumor of the pathologically proven malignant nodes was 0.58 (range, 0.32 to 1.61). Benign nodes had a median ratio of 0.40 (range, 0.21 to 1.10, p = 0.02). The median value was similar for all centers except one. Receiver operating characteristics analysis determined the optimal value of the ratio that maximized sensitivity to be 0.56 or greater (+LR 6.6, sensitivity 94%, specificity 72%). CONCLUSIONS: The ratio of the maxSUV of the mediastinal (N2) lymph node to the maxSUV of the primary tumor in patients with nonsmall-cell lung cancer predicts mediastinal nodal pathology across different PET centers. When the ratio is 0.56 or greater, there is a 94% chance that the node is malignant. The ratio may take into account the different techniques used at different centers. PMID- 17462408 TI - Pneumonectomy for bronchogenic carcinoma: analysis of factors predicting survival. AB - BACKGROUND: The aim of this study was to identify risk factors associated with survival after pneumonectomy for non-small cell lung cancer. METHODS: This was a retrospective study of 155 patients who underwent a pneumonectomy for non-small cell lung cancer at Roswell Park Cancer Institute between 1986 and 2002. Medical record review ascertained information on preoperative assessment including pulmonary function tests and clinical characteristics, postoperative complications, and overall survival. Multivariate Cox proportional hazards models to calculate the hazard ratios and 95% confidence intervals were used. Kaplan Meier cumulative survival curves (with log-rank p values) were generated for selected variables. RESULTS: The median age was 58 years at the time of surgery; 65% of patients were males. Squamous cell carcinoma (54%) and adenocarcinoma (33%) were the predominant histologic types. The median time to relapse was 11 months, and the overall median survival was 15.6 months. An American Society of Anesthesiology score of less than 3, squamous histology, and lower pathologic stage were significant independent predictors of improved survival. Current smoking status (hazard ratio = 1.87, 95% confidence interval: 1.30 to 2.70) and left tumor location (hazard ratio = 1.40, 95% confidence interval: 0.97 to 2.03) were associated with a trend toward poorer survival. Sixty-four patients (41%) had postoperative complications. The operative mortality from pneumonectomy was 9 of 155 (5.8%). CONCLUSIONS: American Society of Anesthesiology score, histology, pathologic stage, smoking status, and location of the tumor were important predictors of survival in this patient sample. Pneumonectomy for non-small cell lung cancer carries an acceptable operative mortality and provides an important survival benefit. PMID- 17462409 TI - Association between surgeon and hospital volume and in-hospital fatalities after lung cancer resections: the experience of an Asian country. AB - BACKGROUND: We used 4-year nationwide population-based data to explore the volume outcome relationships for lung cancer resections in Taiwan and to determine whether there is any association between high-volume hospitals or high-volume surgeons and lower in-hospital mortality rates. METHODS: We use pooled data for the years 2001 through 2004 obtained from the National Health Insurance Research Database in Taiwan. A total of 4,841 patients, identified as having undergone pulmonary resections for lung or bronchial tumors during the period of this study, were treated by 377 surgeons in 79 hospitals. Multivariate logistic regression analyses were then employed to assess the crude and adjusted odds ratio of in-patient fatalities between surgeon and hospital lung cancer resection volume groups. RESULTS: Patients treated by low-volume surgeons had significantly higher in-hospital fatality rates than those treated by either medium-volume surgeons (2.3% versus 1.0%; p < 0.001) or high-volume surgeons (2.3% versus 0.6%; p < 0.001). However, hospital case volume alone is not a significant predictor of hospital in-patient fatalities for lung cancer resections. With increasing surgeon volume, there was a decline in the adjusted odds ratio of hospital in patient deaths. The odds of hospital in-patient deaths for those patients treated by low-volume surgeons were 2.04 times those of medium-volume surgeons, and 2.63 times those of high-volume surgeons. CONCLUSIONS: We conclude that after adjusting for patient, surgeon, and hospital characteristics, an inverse volume outcome relationship does exist for surgeons, but not for hospitals, in Taiwan. PMID- 17462410 TI - Self and parental assessment after minimally invasive repair of pectus excavatum: lasting satisfaction after bar removal. AB - BACKGROUND: Improvement of quality of life by minimally invasive repair of pectus excavatum (MIRPE) has been demonstrated only for the period with implanted pectus bar. The aim of this study was to demonstrate the effects of MIRPE on psychosocial and physical well-being after removal of the pectus bar. METHODS: Forty patients (26 boys and 14 girls; mean age, 17 years; range, 10 to 24 years) were assessed. Follow-up was performed for a mean of 54 months after pectus repair (range, 25 to 73 months). Patients were interviewed at 6 months after MIRPE with the bar in place, and patients and parents were assessed a mean of 23 months after bar removal (range, 2 to 48 months). A single-step questionnaire that evaluates psychosocial and physical well-being was independently used by patients and their parents. RESULTS: There was a high level of persistent satisfaction with MIRPE after bar removal (mean total score = 67; maximal score = 80) and a highly significant correlation between self and external assessment (p < 0.001; Spearman correlation coefficient = 0.77). Analysis of specific and total scores revealed a significant improvement of psychosocial and physical well-being after bar implantation, which persisted up to 4 years after bar removal (p < 0.001). Age and sex had no significant impact on the mean specific and total scores either in patients or in parents. Persistent pectus excavatum was noticed in 1 patient after bar removal (2.5%) as a result of treatment failure. CONCLUSIONS: The positive impact of MIRPE on psychosocial and physical well-being in children and adolescents persists after bar removal. Thus we consider MIRPE to be justified for cosmetic reasons. PMID- 17462411 TI - Invited commentary. PMID- 17462412 TI - Outpatient microthoracoscopic sympathectomy for palmar hyperhidrosis. AB - BACKGROUND: Sympathectomy for palmar hyperhidrosis has been performed for more than 80 years with excellent long-term results. However, several issues are still unresolved: best surgical approach, which level and the number of levels of the sympathetic chain should be divided and the safety of an outpatient procedure. METHODS: We reviewed the records of 205 patients who underwent sympathectomy for hyperhidrosis at our institution from April 2002 through March 2005. Fifty consecutive patients (24%) underwent an outpatient microthoracoscopic sympathectomy of a single level (T2) for palmar hyperhidrosis. Operative reports, medical records, and clinical charts were reviewed to determine the success of a single level sympathectomy, the incidence of postoperative compensatory hyperhidrosis, and the safety of an outpatient sympathectomy procedure. RESULTS: There were 41 women (82%) and 9 men. Median age was 22 years (range, 14 to 62). The surgical technique consisted of single-lumen endotracheal tube general anesthesia, single bilateral access incisions (4 mm), 3-mm, 30-degree thoracoscope, temporary CO2 insufflation, electrocautery nerve division, and no chest drainage. Median operating time was 22 minutes (range, 14 to 50). Sympathectomy consisted of dividing the sympathetic chain at the T2 level and any accessory nerves present. Anatomically, 19 patients (38%) had accessory nerves, right sided in 11, left sided in 5, and bilateral in 3. Two patients (4%) had a chest tube (10F) placed at the time of sympathectomy because of required lysis of apical pleural adhesions. No patient had postoperative bleeding or a wound infection. All patients were discharged the day of surgery. Median follow-up was 15 months (range, 1 to 36). Compensatory hyperhidrosis developed in 6 patients (12%), in 4 with rest and in 2 during exercise, all within 4 weeks. Successful sympathectomy was achieved in 99% of palmar surfaces without recurrence. A single patient experienced persistent sweating of the left hand. Reoperation was successful by dividing level T3. CONCLUSIONS: Single level (T2) microthoracoscopic sympathectomy for palmar hyperhidrosis has a high success rate with a low incidence of compensatory hyperhidrosis. This minimally invasive procedure can be performed safely as an outpatient and should be the preferred treatment for medical refractory palmar hyperhidrosis. PMID- 17462413 TI - Stereolithographic models for surgical planning in congenital heart surgery. AB - PURPOSE: Currently we are exploring the impact of using rapid prototyping techniques for surgical planning and intraoperative orientation during the correction of complex congenital malformation. DESCRIPTION: We studied a patient with a left abnormal subclavian artery and right descending aorta as a rare cause of dyspnea and dysphagia. The patient was examined by magnetic resonance imaging angiography. The image data were visualized and reconstructed. Afterward a replica of the malformation was fabricated using a rapid prototyping machine. In addition, a stereolithographic model of an intracardiac lesion (ventricular septal defect) was fabricated with data obtained from a computed tomographic scan. EVALUATION: Using data derived from a magnetic resonance imaging angiography or computed tomographic scan linked to proprietary software, we were able to create three-dimensional reconstructions of complex vascular pathology and intracardiac lesions. In addition, we fabricated replicas of congenital malformations using a rapid prototyping machine. The models could be sterilized and taken to the operating room for orientation during the corrective surgical procedure. CONCLUSIONS: Stereolithographic replicas are helpful for choosing treatment strategies, surgical planning of corrections, and intraoperative orientation, and as demonstrations on life-like models for the patient. PMID- 17462414 TI - Use of conventional dual chamber pacemakers with custom lead adapters to induce atrial fibrillation or heart failure in dogs. AB - PURPOSE: The purpose of this article is to show how custom "Y"-lead adapters and standard dual-chamber pacemakers can be used to produce pacing paradigms that will lead to stable experimental models of heart failure and atrial fibrillation. DESCRIPTION: With two custom lead adapters we used both ports of two dual-chamber clinical pacemakers to independently apply various pacing paradigms to either the ventricles or the atria of dogs. EVALUATION: Because both ports of the ventricular pacemaker were used to apply stimuli through one lead, the device did not have to be modified to obtain ventricular pacing rates that are sufficiently elevated to lead to tachycardia-induced heart failure. Similarly, simultaneous use of both ports of the atrial pacemaker can be used to apply stimuli through one atrial lead to induce sustained atrial fibrillation. CONCLUSIONS: These techniques facilitate induction of experimental models of heart failure and atrial fibrillation without the need for modification of the clinical pacemaker. PMID- 17462415 TI - Cardiogenic shock: collaboration between cardiac surgery and cardiology subspecialties to bridge to recovery. AB - Acute cardiogenic shock is a lethal condition that results in death from myocardial failure, arrhythmia, or combinations of both. Aggressive medical, surgical, and interventional maneuvers have helped reduce the mortality. For the most advanced cases, ventricular assist devices have been used for persistent shock states. The purpose of this report is to describe the collaboration between cardiac surgery and cardiology subspecialty in an effort to promote native heart recovery in a complex case of cardiogenic shock requiring coronary artery bypass surgery, percutaneous coronary intervention, ventricular ablative therapy, and mechanical cardiac support. PMID- 17462416 TI - Successful use of bivalirudin as anticoagulant for ECMO in a patient with acute HIT. AB - A patient with myocardial failure after repair of an acute type A aortic dissection had acute heparin-induced thrombocytopenia develop during extracorporeal membrane oxygenation. Heparin was discontinued and the anticoagulant was switched to the direct thrombin inhibitor bivalirudin given with a bolus of 0.5 mg/kg followed by a continuous infusion of 0.5 mg/kg/h. Using this protocol, activated clotting time values ranged from 200 to 220 seconds. After prolonged extracorporeal membrane oxygenation support and recovery of left ventricular function, a right ventricular assist device was implanted during extracorporeal membrane oxygenation support with bivalirudin anticoagulation. For this procedure an additional bolus of 0.25 mg/kg bivalirudin was given, and the infusion rate increased to 1 mg/kg/h to achieve activated clotting time values of 300 to 350 seconds. Surgery was successfully performed with moderate intraoperative and postoperative blood loss and transfusion requirements. PMID- 17462417 TI - Right coronary artery with high takeoff. AB - Anomalous origin of the right coronary artery is an extremely rare anomaly. We describe the case of a patient whose right coronary artery was arising from the ascending aorta with high takeoff. We diagnosed the anomaly incidentally during the operation. After transverse aortotomy for aortic valve replacement, we recognized the transection of the right coronary artery. The right coronary artery ostium was located approximately 5 cm above the right sinus of Valsalva. It was showing a complete transmural course. We repaired the right coronary artery by bypassing it with a saphenous vein graft. PMID- 17462418 TI - Forearm muscle necrosis after radial artery harvesting for coronary artery bypass surgery. AB - Donor site complications after radial artery harvesting for coronary revascularization have been primarily wound related and parasthesias. Forearm muscle compartment syndrome has only been reported once. A patient underwent successful coronary revascularization with a radial artery graft; however his forearm extensor muscles necrosed, possibly from a missed compartment syndrome, and he required tendon transfers to provide finger and wrist extension. PMID- 17462419 TI - Thoracoscopic radiofrequency pulmonary vein isolation and atrial appendage occlusion. AB - A 46-year-old woman with a 7-year history of profoundly symptomatic daily paroxysmal atrial fibrillation had undergone two percutaneous catheter ablations and multiple medication trials. With informed consent, bilateral pulmonary vein isolation and left atrial appendage occlusion as well as ablation of ganglionated plexi were performed by a totally thoracoscopic technique employing a bipolar radiofrequency device. She was discharged home on postoperative day 3 and had one brief episode of atrial fibrillation 1 week later, but has had no further atrial fibrillation for more than 6 months since the procedure. PMID- 17462420 TI - Cardiac resynchronization therapy for mitral systolic anterior motion in a child. AB - We describe the successful use of cardiac resynchronization therapy for treatment of mitral valve systolic anterior motion with left ventricle outflow tract obstruction after re-excision of a subaortic membrane and septal myectomy in a 12 year-old child. In the recovery phase, a total atrioventricular block persisted. Therefore a permanent atrioventricular pacing system was implanted. PMID- 17462421 TI - Bacteremia causes mycotic aneurysm of the aortic arch in 110 days. AB - We report a surgical case of mycotic aneurysm of the aortic arch, the possible cause, and the growth of which were retrospectively determined. A 61-year-old man with diabetes mellitus, free of aneurysm had bacteremia develop that was caused by Staphylococcus aureus. A computed tomographic scan was taken 110 days afterward that revealed an aneurysm in his aortic arch. Total arch replacement was performed, and S. aureus was detected in the aneurysm content. We conjectured that bacteremia caused mycotic aneurysm by day 110. The results of intraoperative microscopic examination also led the authors to decide on a regime for the immediate systemic administration of antibiotics. PMID- 17462422 TI - Takayasu's arteritis mimicking acute aortic dissection. AB - A 17-year-old girl presented with a history of dyspnea on exertion and fever of 1 week duration. She was evaluated elsewhere with transesophageal echocardiography and helical computed tomographic scan, and she had been diagnosed with an acute type I dissection of the aorta. She had also been diagnosed with severe aortic regurgitation and a suspected aortic root abscess. On the operating table, we found no evidence of dissection, but we did find that her aorta was severely thickened and inflamed. The patient's aortic valve was replaced. In view of the left main stem ostial stenosis, we harvested and grafted the left internal thoracic artery to the left anterior descending artery. During the operation it is of paramount importance to rule out dissections involving the arch and coronary ostial narrowing. PMID- 17462423 TI - Staged therapeutic approach in spontaneous coronary dissection. AB - Spontaneous coronary artery dissection is an extremely rare event occurring especially in pregnant women, either peripartum or postpartum. Urgent coronary angiography has to be performed to confirm the diagnosis and to determine the appropriate therapeutic strategy. We present the case of a 36-year-old woman in week 36 of her pregnancy who was admitted in cardiogenic shock due to an acute anterolateral myocardial infarction as a result of spontaneous coronary artery dissection. While she was on aortic balloon counterpulsation the patient underwent a cesarean section and gave birth to a healthy child. Subsequently she successfully underwent off-pump coronary artery bypass surgery. PMID- 17462424 TI - Successful resection of a primary left ventricular schwannoma. AB - Nerve sheath neoplasm of the heart is rare. We report the case of a patient with a giant schwannoma of unique ventricular origin. Resection of the schwannoma and subsequent coronary reconstruction were required. PMID- 17462425 TI - Cardiac paraganglioma presenting with acute myocardial infarction and stroke. AB - We report an unusual presentation of cardiac paraganglioma with acute myocardial infarction and stroke induced by exercise and review the literature regarding this rare cardiac tumor. PMID- 17462426 TI - Treatment of plastic bronchitis. AB - Plastic bronchitis may be seen after palliative surgery for cyanotic heart disease. Although type II (acellular) casts are seen more commonly, we describe a type I cast after palliative surgery for cyanotic heart disease in which ligation of thoracic duct did not result in complete resolution. PMID- 17462427 TI - Reverse Blalock-Taussig shunt facilitates the growth of the ascending aorta after hybrid palliation. AB - A 13-day-old baby girl with tricuspid atresia (IIc), who was prematurely born at 32 weeks and 5 days of gestation and weighed 2.2 kg, underwent bilateral pulmonary artery banding, ductal stenting, and reverse Blalock-Taussig shunt. Cardiac computerized tomography at 4 months postoperatively showed that the ascending aorta outgrew the somatic growth, presumably thanks to the forward flow through the reverse Blalock-Taussig shunt. At 6 months postoperatively, the patient underwent a successful second-stage operation. PMID- 17462428 TI - Methicillin-resistant Staphylococcus aureus pneumonia after thoracic surgery: successful treatment with linezolid after failed vancomycin therapy. AB - We describe 3 patients with methicillin-resistant Staphylococcus aureus, hospital acquired pneumonia who demonstrated a microbiological and clinical failure with vancomycin despite adequate trough levels. All 3 patients were cured with linezolid. PMID- 17462429 TI - Graft pneumonectomy and delayed contralateral lung transplantation. AB - We believe that contralateral single lung transplantation after graft pneumonectomy has not been reported yet in the literature. We present the case of an emphysematous patient who received a unilateral left lung transplant and had severe stenosis in bronchial anastomosis and bronchiectasis develop. Four years after transplantation we decided to perform a left pneumonectomy and a delayed right lung transplantation. Nine months after the procedure the patient is ambulatory and is not dependent on oxygen support. PMID- 17462430 TI - Prolonged survival in a patient with recurrent pulmonary metastases secondary to mucinous cystadenocarcinoma of the appendix with pseudomyxomatous peritonei. AB - We report a 65-year-old man presenting with recurrent pulmonary metastases 20 years after an appendectomy for mucinous cystadenocarcinoma with pseudomyxomatous peritonei. He underwent bilateral staged metastatectomies for metastases 7 years after the diagnosis and further metastasectomy after a recent recurrence. This is a rare case of recurrent pulmonary metastatic mucinous cystadenocarcinoma, and despite poor prognosis and nondefinitive initial treatment, this patient remains alive and well 20 years later. PMID- 17462431 TI - Elastofibroma dorsi: clinicopathologic review of 6 cases. AB - Elastofibroma dorsi is a rare, benign lesion arising from connective tissue and usually found at the angle of the scapula. Surgical resection is often indicated in the presence of an enlarging mass or when malignancy can not be excluded. Herein we report our most recent case of elastofibroma dorsi and our review of 6 cases from the past 16 years. PMID- 17462432 TI - Tracheal stenting of iatrogenic tracheal injury: a novel management approach. AB - We report the case of a patient who had an intubation-related tracheal injury who we treated by deployment of a covered tracheal stent. This approach may be preferable to other alternatives in patients with a prohibitive risk of mortality with surgical repair or in an injury with sequelae not suitable for conservative management. PMID- 17462433 TI - Congenital sternal cleft in an adult male not associated with cardiac defects or ectopia cordis. PMID- 17462434 TI - Papillary muscle infarction: the role of magnetic resonance imaging. PMID- 17462435 TI - Simple, safe and easy technique to ensure the correct length of artificial chordae in mitral valve repair. AB - Replacement of diseased chordae with Gore-Tex sutures (W. L. Gore & Assoc, Flagstaff, AZ) in patients with degenerative mitral valve insufficiency has become a standard technique used by surgeons in mitral valve repair with good long-term results. Nevertheless, determining the correct length of the artificial chordae has remained problematic. Although various procedures have been previously published, in this article we describe our approach used to achieve an accurate chordal height adjustment. PMID- 17462436 TI - Transaortic delivery of the transmitral lesion in a complete maze procedure. AB - A 68-year-old hypertensive diabetic woman with chronic atrial fibrillation presented with progressive congestive symptomatology. She was diagnosed with severe aortic stenosis, moderate mitral regurgitation, and critical right coronary artery stenosis. In addition to coronary revascularization and bioprosthetic aortic valve replacement, she underwent a mitral valve repair and a complete cryoMaze procedure through a transaortic approach. This technique obviates a separate left atriotomy for the mitral repair and Maze procedure. It affords excellent exposure, while reducing cross clamp and cardiopulmonary bypass time as well as avoiding the potential sequelae of bleeding and traction injuries resulting from a left atriotomy. PMID- 17462437 TI - A "double overlap" suture technique for the proximal attachment of a composite graft to the aortic annulus. AB - Repair of an aortic root aneurysm using a composite graft is sometimes complicated by proximal suture line bleeding, which may be very difficult to control. We adopted a previously described technique of "double overlap" sutures on the annulus and the prosthetic cuff, which has virtually eliminated this complication. PMID- 17462438 TI - New technique of aortic root reconstruction with aortic valve annuloplasty in ascending aortic aneurysm. AB - We describe a new technique of aortic valve conservation for ascending aortic aneurysm with aortic valvular insufficiency. This technique allows a total anatomic aortic root reconstruction associated with an aortic annuloplasty preventing late annulus dilation and reoperation. Preliminary results demonstrate the feasibility and the safety of this new original procedure. PMID- 17462439 TI - Total arterial coronary revascularization using arterial bypass circle with multiple inflows. AB - Coronary artery bypass techniques, currently applied to maximize the benefits of multiple arterial coronary conduits, render the newly constructed myocardial flow dependent on a single source "inflow" of blood. We describe a technique for total arterial coronary revascularization with multiple inflows; the distal end of the pedicled right internal thoracic artery is anastomosed to the distal end of a free radial artery, and the other end of the radial artery is then connected to the ascending aorta. This vascular circle, passed in a retro-cardiac fashion, is used to revascularize the inferio-lateral surface of the heart using multiple side-to-side anastomoses. The "sacred" left internal thoracic artery is reserved to revascularize the anterior wall of the myocardium, independent of the arterial circle. PMID- 17462440 TI - Convenient and improved method to distinguish the intersegmental plane in pulmonary segmentectomy using a butterfly needle. AB - In the traditional method of segmentectomy, the plane between segments where removal is to occur is demarcated by inflating the normal lung, while keeping the segment to be removed airless. Our method, the opposite of convention, involves inflating only the involved segment by instilling oxygen through a butterfly needle into the bronchus subtending the segment. This saves time and therefore benefits the patient. PMID- 17462442 TI - Use of the STS database by insurance companies. PMID- 17462444 TI - The mechanisms of chronic ischemic mitral regurgitation. PMID- 17462446 TI - Can cryopreservation destroy the extracellular matrix of pulmonary allografts? PMID- 17462448 TI - Are cerebral emboli real when perfluorocarbon emulsion (AF0144) is used in cardiac surgery? PMID- 17462449 TI - Alternative conservative therapy. PMID- 17462452 TI - Management of postintubation tracheobronchial ruptures. PMID- 17462454 TI - Perioperative blood transfusion and blood conservation in cardiac surgery: the Society of Thoracic Surgeons and The Society of Cardiovascular Anesthesiologists clinical practice guideline. AB - BACKGROUND: A minority of patients having cardiac procedures (15% to 20%) consume more than 80% of the blood products transfused at operation. Blood must be viewed as a scarce resource that carries risks and benefits. A careful review of available evidence can provide guidelines to allocate this valuable resource and improve patient outcomes. METHODS: We reviewed all available published evidence related to blood conservation during cardiac operations, including randomized controlled trials, published observational information, and case reports. Conventional methods identified the level of evidence available for each of the blood conservation interventions. After considering the level of evidence, recommendations were made regarding each intervention using the American Heart Association/American College of Cardiology classification scheme. RESULTS: Review of published reports identified a high-risk profile associated with increased postoperative blood transfusion. Six variables stand out as important indicators of risk: (1) advanced age, (2) low preoperative red blood cell volume (preoperative anemia or small body size), (3) preoperative antiplatelet or antithrombotic drugs, (4) reoperative or complex procedures, (5) emergency operations, and (6) noncardiac patient comorbidities. Careful review revealed preoperative and perioperative interventions that are likely to reduce bleeding and postoperative blood transfusion. Preoperative interventions that are likely to reduce blood transfusion include identification of high-risk patients who should receive all available preoperative and perioperative blood conservation interventions and limitation of antithrombotic drugs. Perioperative blood conservation interventions include use of antifibrinolytic drugs, selective use of off-pump coronary artery bypass graft surgery, routine use of a cell-saving device, and implementation of appropriate transfusion indications. An important intervention is application of a multimodality blood conservation program that is institution based, accepted by all health care providers, and that involves well thought out transfusion algorithms to guide transfusion decisions. CONCLUSIONS: Based on available evidence, institution-specific protocols should screen for high-risk patients, as blood conservation interventions are likely to be most productive for this high-risk subset. Available evidence-based blood conservation techniques include (1) drugs that increase preoperative blood volume (eg, erythropoietin) or decrease postoperative bleeding (eg, antifibrinolytics), (2) devices that conserve blood (eg, intraoperative blood salvage and blood sparing interventions), (3) interventions that protect the patient's own blood from the stress of operation (eg, autologous predonation and normovolemic hemodilution), (4) consensus, institution-specific blood transfusion algorithms supplemented with point-of-care testing, and most importantly, (5) a multimodality approach to blood conservation combining all of the above. PMID- 17462455 TI - The gut is a motor of organ system dysfunction. PMID- 17462456 TI - The effect of neoadjuvant chemotherapy on lymph node micrometastases in squamous cell carcinomas of the thoracic esophagus. AB - BACKGROUND: Neoadjuvant chemotherapy (NACT) has been postulated but not yet proven to eradicate micrometastases and improve the prognosis of patients with advanced esophageal squamous cell carcinomas (ESCC). Cytokeratin immunohistochemistry of the lymph nodes of ESCC revealed immunohistochemical micrometastases (IHM) and cytokeratin deposits (CD), which are hyalinized denucleated particles considered to be cadavers of carcinoma cells. Successful chemotherapy should convert cancer cells from IHM to CD and improve the status of ESCC patients from systemic disease to regional disease. METHODS: Cytokeratin immunostaining of surgically removed lymph nodes was performed for 107 patients with node-positive ESCC, including 32 patients without preoperative treatment (Surgery group) and 75 patients undergoing NACT using CDDP, doxorubicin hydrochroride, and 5-fluorouracil (NACT group). Cytokeratin-positive staining was done for serial hematoxylin-eosin-stained sections and classified as pathologic metastasis, IHM, or CD. RESULTS: CD was observed less frequently in the Surgery group than in the NACT group (6% vs 43%, P < .0001), whereas IHM was more frequent in the former (47% vs 24%, P = .019). IHM was a poor prognostic factor in both groups, whereas CD was a favorable one in the NACT group. The effect of chemotherapy on IHM was classified as eradicated, IHM(-)/CD(+); persistent, IHM(+)/CD(+); no effect, IHM(+)/CD(-); or not informative, IHM(-)/CD(-). This classification correlated well with the clinical response of the primary neoplasm, number of pathologic metastases, and postoperative survival (3-year survival rates: 78%, 18%, 0%, and 38%). IHM/CD was found to be an independent prognostic factor together with the number of pathologic metastases in the multivariate analysis. CONCLUSIONS: Disappearance of IHM and the emergence of CD suggest the eradication of micrometastases by NACT. The clinical benefit of NACT was apparent for IHM(-)/CD(+) patients with node-positive ESCC. PMID- 17462457 TI - Combined vascular resection in operative resection for hilar cholangiocarcinoma: does it work or not? AB - BACKGROUND: It is still not clear how combined vascular resection affects the outcome of patients with hilar cholangiocarcinoma. Our aim was to evaluate implications of combined vascular resection in patients with hilar cholangiocarcinoma by analyzing the outcomes of all patients who underwent operative resection. METHODS: A total of 161 of 228 consecutive patients with hilar cholangiocarcinoma underwent bile duct resection with various types of hepatectomy (88%) and pancreaticoduodenectomy (4%). Combined vascular resection was carried out in 43 patients. Thirty-four patients had portal vein resection alone, 7 patients had both portal vein and hepatic artery resection, and 2 patients had right hepatic artery resection only. The outcomes were compared between the 3 groups: the portal vein resection alone (34), hepatic artery resection (9), and non-vascular resection (118). RESULTS: Histologically-positive tumor invasion to the portal vein beyond the adventitia was present in 80% of 44 patients undergoing combined portal vein resection. Operative mortality occurred in 11 (7%) patients. The survival rates of the non-vascular resection group were better than that of the portal vein resection alone and the hepatic artery resection groups: 1, 3, and 5 years after curative resection, 72%, 52%, and 41% versus 47%, 31%, and 25% (P < .05), and 17%, 0%, and 0% (P < .0001), respectively. Multivariate analysis showed 4 independent prognostic factors of adverse effect on survival after operation; operative curability, lymph node metastases, portal vein resection, and hepatic artery resection. CONCLUSIONS: Although both portal vein and hepatic artery resection are independent poor prognostic factors after curative operative resection of locally advanced hilar cholangiocarcinoma, portal vein resection is acceptable from an operative risk perspective and might improve the prognosis in the selected patients, however, combined hepatic artery resection can not be justified. PMID- 17462458 TI - Repeat hepatectomy for recurrent hepatocellular carcinoma. AB - BACKGROUND: Long-term prognosis of patients with hepatocellular carcinoma (HCC) after partial hepatectomy remains unsatisfactory because of the high incidence of recurrence in the liver remnant. Controversy exists about the efficacy of repeat hepatectomy for recurrent HCC patients. The purpose of this study was to retrospectively examine and clarify the significance of repeat hepatectomy in the treatment of recurrent HCC. METHODS: From January 1990 to December 2004, 84 patients with recurrent HCC underwent a second hepatectomy with curative intent. Survival rates in these 84 patients were analyzed retrospectively. RESULTS: After the second hepatectomy, the overall 5-year survival rate was 50% for the 84 patients included in this study; the corresponding recurrence-free survival rate was 10%. Multivariate analysis showed that the second hepatectomy performed between 1997 and 2004 (P < .001) and the absence of microscopic vascular invasion at the second hepatectomy (P = .001) were the significant and independent prognostic factors for overall survival after the second hepatectomy. The overall 5-year survival rate after the second hepatectomy was 80% in 46 patients who had both these prognostic factors. However, even in the subgroup with good long-term survival, the 5-year recurrence-free survival rate was only 6%. The more times hepatectomy was repeated, the shorter the recurrence-free interval became. CONCLUSIONS: Repeat hepatectomy for recurrent HCC had survival benefits, especially for patients without microscopic vascular invasion. However, the incidence of re-recurrence after the second hepatectomy was high, and the recurrence-free interval was short, even in the subgroup with survival benefits. The effectiveness of repeat hepatectomy for curing recurrent HCC is limited. PMID- 17462459 TI - A revised scoring system utilizing serum alphafetoprotein levels to expand candidates for living donor transplantation in hepatocellular carcinoma. AB - BACKGROUND: The development of living donor liver transplantation has stimulated discussion about the expansion of tumor burden limits for patients with hepatocellular carcinoma (HCC). Although serum alphafetoprotein (AFP) level is an important predictor of tumor recurrence, it is not included in the existing selection criteria for HCC in transplantation. METHODS: We performed a retrospective study of 63 consecutive adults with HCC diagnosed preoperatively who received living donor liver transplantation from February 1999 to September 2005 and survived over 1 month. The authors devised new scoring criteria that included tumor size, tumor number, and pretransplant AFP level as prognostic factors. The score of each parameter was classified from 1 to 4 points (tumor size, < or =3, 3.1 to 5, 5.1 to 6.5, >6.5 cm; tumor number, 1, 2 or 3, 4 or 5, or > or =6 nodules; and AFP, < or =20, 20.1 to 200, 200.1 to 1000, >1000 ng/mL, respectively). We defined that 3 to 6 points and 7 to 12 points were "transplantable" and "nontransplantable," respectively. The usefulness of the devised criteria was then investigated as a method of selecting candidates with HCC for transplantation. RESULTS: The candidates' overall 3-year survival rate and recurrence-free survival rate were 67% and 70% after transplantation, respectively. Based on pretransplant imaging, 37 (59%), 41 (65%), and 44 (70%) of the 63 patients met the Milan criteria, University of Californica, San Francisco (UCSF) criteria, and the new scoring criteria. Their 3-year survival rates were 80%, 78%, and 79%, respectively. Moreover, based on posttransplant data, the scoring criteria correlated with the risk of death and HCC recurrence (Milan criteria, P = .005 and .001; UCSF criteria, P = .013 and .001 for death and recurrence; scoring criteria, P < .001 for both). CONCLUSIONS: The newly devised scoring criteria could expand usefully current selection criteria for transplantation without detrimentally affecting outcome in the living donor transplantation setting for HCC. PMID- 17462460 TI - Prognostic relevance of lymph node ratio following pancreaticoduodenectomy for pancreatic cancer. AB - BACKGROUND: The presence or absence of lymph node metastases is known to be an important prognostic factor for patients with pancreatic cancer. Few studies have investigated the ratio of the number of lymph nodes harboring metastatic cancer to the total number of lymph nodes examined (lymph node ratio [LNR]) with regard to outcome after pancreaticoduodenectomy for ductal cancer of the pancreas. METHODS: Between 1995 and 2005, a total of 905 patients underwent pancreaticoduodenectomy for pancreatic adenocarcinoma. Demographics, operative data, number of lymph nodes evaluated, number of lymph nodes with metastatic carcinoma, LNR, pathologic margin status, and long-term survival were analyzed. RESULTS: There were 187 (20.7%) of the 905 patients who had negative peripancreatic lymph nodes (N0), whereas 718 (79.3%) of the 905 patients had lymph node metastases (N1). The median number of lymph nodes evaluated in the N0 group was 15 versus 18 in the N1 group (P = .12). At median follow-up of 24 months, the median survival for all patients was 17.4 months, and the 5-year actuarial survival rate was 16.1%. Patients with lymph node metastases had a shorter median overall survival (16.5 months) compared with patients with negative lymph nodes (25.3 months; P = .001). Compared with the total number of lymph nodes examined or total number of lymph node metastases, LNR was the most compelling predictor of survival. As the LNR increased, median overall survival decreased (LNR = 0, 25.3 months; LNR > 0 to 0.2, 21.7 months; LNR > 0.2 to 0.4, 15.3 months; LNR > 0.4, 12.2 months; P = .001). After adjusting for other factors associated with survival, LNR remained an independent predictor of overall survival (P < .001). CONCLUSIONS: After pancreaticoduodenectomy for adenocarcinoma of the pancreas, LNR was one of the most powerful predictors of survival. LNR should be considered when stratifying patients in future clinical trials. PMID- 17462461 TI - Distal pancreatectomy with splenic preservation revisited. AB - BACKGROUND: Splenic preservation (SP) during distal pancreatectomy can be accomplished by ligating the main splenic artery and vein relying on blood supply from the short gastric vessels. The purpose of this study was to examine the short-term implications of this operation, comparing it to the outcomes following distal pancreatectomy with splenectomy. METHODS: The records of 259 patients who underwent distal pancreatectomy with and without SP at Massachusetts General Hospital from 1994 to 2004 were reviewed. RESULTS: A total of 29% of patients underwent SP with this technique. These patients were more likely to be women (74% vs 56%, P = .008) and to have benign disease (93% vs 54%, P < .0001). Their operative times were shorter (2.5 vs 3.1 h, P < .0001), they had less blood loss (300 vs 500 ml, P < .0001) and a shorter duration of stay (6 days [interquartile range, 5 to 7] vs 7 days [interquartile range, 5 to 8], P = .001). SP was not a significant predictor of complications in either univariate (P = .445) or adjusted analysis (P = .543). One patient (1.4%) in the SP group was reoperated for splenic infarction and two patients (1.1%) in the splenectomy group for abscess and hemorrhage. There were 2 (0.8%) postoperative deaths, both in the splenectomy group. CONCLUSIONS: Splenic preservation relying on blood supply from the short gastric vessels is reliable and safe and does not have a higher incidence of postoperative complications when compared to traditional distal pancreatectomy with splenectomy. The current series validates this approach and provides further evidence of its feasibility and safety. PMID- 17462462 TI - Cervical thymectomy for intrathymic parathyroid adenomas during minimally invasive parathyroidectomy. AB - BACKGROUND: The development of an intrathymic parathyroid adenoma is common, and thymectomy is a significant component of the parathyroid surgeon's technical armamentarium. Over the last decade, minimally invasive parathyroidectomy (MIP) has become the standard technique for removal of an abnormal parathyroid gland, and the requirement for thymectomy should remain unchanged during the era of minimally invasive techniques. The aim of this paper was to assess the feasibility and outcomes of cervical thymectomy for intrathymic parathyroid adenomas during MIP. METHODS: This is a retrospective case series. The study group comprised all patients undergoing parathyroidectomy in the University of Sydney Endocrine Surgical Unit during a 5-year period (January 2001 to December 2005). Patients undergoing MIP and open parathyroidectomy with a concomitant cervical thymectomy were compared. RESULTS: A total of 840 patients underwent parathyroid surgery for primary hyperparathyroidism (PHPT) during this period. A total of 30 MIP procedures with concurrent thymectomy were performed, and 99 open bilateral neck explorations with cervical thymectomy were performed. Of the MIP thymectomy group, there were 25 female and 5 male patients; the average age was 57 years (range, 22 to 82). A mean length of 34 mm of thymus was extracted via the minimally invasive approach (range, 8 to 85 mm). In 5 cases, only fatty tissue was identified histologically, and, in 5 cases, a small supernumerary parathyroid gland was identified in the histologic specimen. Only 1 patient suffered temporary, recurrent laryngeal nerve palsy; there were no cases of postoperative hemorrhage requiring return to the operating room. CONCLUSIONS: Cervical thymectomy for removal of intrathymic parathyroid adenomas can be performed during lateral focused mini-incision MIP with a safety and efficacy equivalent to open bilateral neck explorations. PMID- 17462463 TI - Effects of fresh versus banked blood transfusions on microcirculatory hemodynamics and tissue oxygenation in the rat cremaster model. AB - BACKGROUND: Blood transfusion is recommended to prevent the harmful effects of tissue hypoxia. Whether transfusion alleviates hypoxia at the microcirculatory level is unknown. The objective of this study was to determine whether the effects of fresh blood transfusion on microcirculatory hemodynamics after blood loss are preferable to the effects of banked blood transfusion. METHODS: Intravital microcirculatory hemodynamics was observed in the rat cremaster muscle flap by observers who were not blinded. Lewis rats were divided into 6 groups (8 rats per group): (1) untreated control; (2) blood withdrawal only (1 mL); (3) blood withdrawal and transfusion of rat fresh blood (1 mL); (4) blood withdrawal and transfusion with 1-day rat banked blood (1 mL); (5) blood withdrawal and transfusion with 1-week rat banked blood (1 mL); and (6) blood withdrawal and transfusion with 2-week rat banked blood (1 mL). Red blood cell velocity, blood flow, vessel diameter, functional capillary perfusion, red blood cell deformability, leukocytes (granulocytes and lymphocytes), and tissue oxygenation levels were monitored before transfusion and for 5 hours afterward. Histology was performed to evaluate tissue inflammation (hematoxylin and eosin [H&E] stain and myeloperoxidase [MPO] immunoassay) and hypoxia (Hypoxyprobe assay). RESULTS: Fresh- and banked blood-transfused groups did not differ in vessel diameters or red blood cell velocities. Functional capillary density was greater in the fresh blood group when compared to 1- and 2-week banked blood-transfused groups. The overall number of granulocytes (rolling, sticking, and transmigrated) was greater in all groups after transfusion (P < .001). Rats transfused with banked blood had a greater number of rolling lymphocytes and more inflammation. Tissue oxygenation levels were improved after fresh blood transfusion in comparison to banked blood (9.5 mm Hg vs 8 mm Hg, P = .02). CONCLUSIONS: Fresh blood transfusion is more effective in relieving effects of microcirculatory hypoxia. Banked blood, in particular 2-week stored blood, has limited capacity of improving tissue oxygenation. PMID- 17462464 TI - Laparoscopic subtotal colectomy for acute or severe colitis complicating inflammatory bowel disease: a case-matched study in 88 patients. AB - BACKGROUND: The aim of this study was to assess the morbidity of laparoscopic subtotal colectomy (STC) with or without anastomosis in patients with acute or severe colitis (SAC) complicating inflammatory bowel disease (IBD) who failed medical treatment. METHODS: Forty patients undergoing laparoscopic STC for SAC complicating IBD were identified and well-matched for age, gender, ASA score, and IBD severity at the time of colectomy (acute colitis vs steroid dependence only) with 48 patients undergoing open STC. RESULTS: There was no operative mortality. Mean (+/-SD) operative time was similar after laparoscopic and open STC (253 +/- 56 vs 231 +/- 75 min; NS). Two patients (5%) required conversion into laparotomy due to intensive adhesions (n = 1) and colonic fistula (n = 1). Overall morbidity and hospital stay was similar after laparoscopic STC and open STC (35% vs 56%) (9 +/- 3 vs 12 +/- 7 days) (P > .1) respectfully. After laparoscopic STC, 84% of the patients underwent restorative intestinal continuity (with either ileorectal or ileoanal anastomosis) through reoperative laparoscopy (n = 15) or elective incision at the site of previous stoma (n = 16). CONCLUSIONS: This case-matched study suggests that laparoscopic STC was as safe and effective as open STC for IBD patients with SAC. A laparoscopic STC allows restoration of intestinal continuity restoration (ie, ileal pouch anal or ileorectal anastomosis) through a laparoscopic approach or elective incision for the majority of the patients. For these reasons, laparoscopic approach represents the best approach for colitis complicating IBD. PMID- 17462465 TI - Association between operative closure type and acute infection, local recurrence, and disease surveillance in patients undergoing breast conserving therapy for early-stage breast cancer. AB - BACKGROUND: This study investigates the effect of full-thickness versus superficial closure of the breast parenchyma on the likelihood of subsequent infection and local recurrence after lumpectomy for early-stage breast cancer. In patients undergoing breast-conserving therapy (BCT), operative closure technique has been largely influenced by expected cosmetic outcome. However, the common practice of promoting postoperative fluid collection raises concerns about potential bacterial colonization, tumor cell migration, and impaired post-BCT surveillance. METHODS: From 1985 through 2004, operative closure technique was determined in 516 breasts in 580 women with stage T0-2N0-1 breast cancers undergoing BCT. Medical records were reviewed to determine closure technique, incidence of postoperative infection, and local recurrence characteristics. RESULTS: Median follow-up was 6.4 years from the completion of radiotherapy. The rate of acute infection was higher with the superficial closure technique: 11.7% (27/230) versus 5.2% (15/286) (P = .009). In T1-2 patients, there was no difference in the rate of local recurrence based on closure type: 5.6% (11/195) versus 3.5% (8/231) (P = .348). On multivariate analysis, acute infections and margin status were associated with increased local recurrence. Superficial closure was associated with larger recurrences less likely to be detected on mammogram. In stage T0-T2 patients, 80% of recurrent tumors after superficial closures were greater than 1 cm compared with no recurrent tumors greater than 1 cm after full-thickness closures (P = .005). In patients with superficial closure, 29% of recurrences in the tumor bed were initially detected on mammogram versus 100% in patients with deep closure (P = .003). CONCLUSIONS: Closure method was not predictive of local recurrence. Our findings regarding infection and post treatment surveillance suggest, however, that full-thickness closure may be the preferred technique in BCT patients. PMID- 17462466 TI - High dietary carbohydrates decrease gallbladder volume and enhance cholesterol crystal formation. AB - BACKGROUND: Animal and human data suggest that a diet high in refined carbohydrates leads to gallstone formation. However, no data are available on the role of dietary carbohydrates on gallbladder volume or on cholesterol crystal formation. Therefore, we tested the hypothesis that a high carbohydrate diet would alter gallbladder volume and enhance cholesterol crystal formation. METHODS: At 8 weeks of age, 60 lean and 36 obese leptin-deficient female mice were fed a 45% carbohydrate diet while an equal number of lean and obese mice were fed a 75% carbohydrate diet for 4 weeks. All animals then underwent cholecystectomy, and gallbladder bile volume was recorded. Bile was pooled, filtered, and maintained in a water bath at 37 degrees C for 14 days. Birefringent cholesterol crystals in bile were counted daily; crystal observation time and crystal mass were determined. RESULTS: The crystal observation time was significantly shortened in both lean and obese mice on the 75% diet compared with their counterparts on the 45% diet. The crystal mass was significantly increased in the lean mice on the 75% diet compared with the 45% diet. Gallbladder volumes were significantly reduced in both lean and obese mice on the 75% diet compared with their counterparts on the 45% diet. CONCLUSIONS: These data suggest that a high carbohydrate diet decreases gallbladder volume, shortens cholesterol crystal observation time, and increases crystal mass. We conclude that dietary carbohydrates may play a role in cholesterol gallstone formation by altering biliary motility and by enhancing crystal formation. PMID- 17462467 TI - Dynamics of albumin synthesis after major rectal operation. AB - BACKGROUND: Major abdominal operations were found to be associated with long lasting metabolic changes, such as accelerated release of stress hormones and carbohydrate turnover. It is unknown currently whether acute changes of hepatic protein metabolism persist in a similar way. We wanted to determine the long-term dynamics of albumin synthesis and its relationship to whole body protein breakdown and albumin concentration after major rectal operations. METHODS: We used stable isotope tracer techniques to determine albumin synthesis and whole body protein breakdown (rate of appearance of leucine, Ra) in postoperative patients about 1 week after low anterior rectal resection and also during convalescence (about 4 months after operation), and in healthy controls. Consecutive blood sampling was carried out during continuous isotope infusion (1 [(13)C]-leucine, 0.16 micromol/kg min). RESULTS: Serum albumin concentrations were close to the lower normal limit in patients early after operation but were comparable to controls in convalescent patients. Simultaneously, albumin synthesis was increased in the early postoperative phase (0.53 +/- 0.0.5%/h) compared with convalescent patients (0.32 +/- 0.04) and controls (0.28 +/- 0.04) (P < .01 each). A significant inverse correlation could be found between plasma albumin concentration and corresponding rates of albumin synthesis. Early after operation patients showed an increased leucine Ra (3.25 +/- 0.23 micromol/kg min) that was greater than that of convalescent patients (2.37 +/- 0.06 micromol/kg min, P < .05). Leucine Ra in both patient groups were greater than the rates in controls (2.01 +/- 0.07 micromol/kg min, P < .01) Albumin synthesis correlated weakly with whole body protein breakdown rate. CONCLUSIONS: Albumin synthesis and total body protein breakdown are increased after major abdominal operation, but albumin synthesis returns to control values only during convalescence. Hypoalbuminemia after rectal operations may be associated with high rates of albumin synthesis and is, therefore, not necessarily an indicator of insufficient hepatic function or poor nutritional status in that particular situation. PMID- 17462468 TI - Donor pretreatment with FK506 reduces reperfusion injury and accelerates intestinal graft recovery in rats. AB - BACKGROUND: FK506 alleviates warm ischemia-reperfusion injury, but it remains unknown if such protection is manifest after cold storage and transplantation. We studied the early outcome after transplantation of intestines from donors pretreated with FK506 compared to grafts from controls treated with saline (154 mM NaCl). METHODS: Sprague-Dawley rats received 0.3 mg/kg FK506 or saline intravenously 6 hours before graft retrieval. The small bowel was harvested, stored for 3 hours, and then transplanted heterotopically. Samples were taken after preservation and at 20 minutes, 6 hours, 12 hours, and 24 hours after reperfusion. Heat shock protein 72 (Hsp72) and iintercellular adhesion molecule (ICAM)-1 expression and nuclear factor kappaB (NF-kappaB) activation were assessed via Western blots and eelectrophoretic mobility shift assay (EMSA), respectively. Dissacharidase activity and enterocyte proliferation rate were also studied. RESULTS: Preservation injury was similar between groups, but pretreated grafts had better morphology already 20 minutes after reperfusion. Control grafts always had thinner mucosa and more PMN infiltration. Hsp72 expression was greater in pretreated grafts. ICAM-1 was absent after harvesting, preservation, and immediately after reperfusion but increased in control grafts at the later time points. Control grafts showed a biphasic NF-kappaB activation pattern, whereas NF kappaB activation was inhibited effectively in pretreated grafts. Dissacharidase activity decreased during the first 6 hours after reperfusion but recovered within 24 hours in pretreated grafts but not in control grafts. Earlier enterocyte proliferation was observed in pretreated grafts. CONCLUSIONS: FK506 donor pretreatment reduced graft proinflammatory activation and neutrophil inflammation. Pretreated groups revealed a milder reperfusion injury and accelerated morphologic and functional recovery. The mechanisms involved appear to involve Hsp72 upregulation and NF-kappaB inhibition. PMID- 17462469 TI - Cross-linked poly (gamma-glutamic acid) attenuates peritoneal adhesion in a rat model. AB - BACKGROUND: Poly (gamma-glutamic acid) (PGA) is a naturally occurring biodegradable polymer produced by Bacillus subtilis. PGA is crosslinked by gamma irradiation to yield crosslinked PGA (XL). XL absorbs large amounts of fluid and forms a biodegradable viscous hydrogel. In the present study, we evaluated the anti-adhesive effect of XL in a rat abdominal wall defect and cecal abrasion model. METHODS: Abdominal wall resection and cecal abrasion were carried out in rats (abrasion-no-treatment group, n = 12). In the treatment groups, the following materials were applied: PGA (n = 12), XL (n = 16), hyaluronic acid (HA) (n = 12), Seprafilm (n = 12), and Interceed (n = 10). A week later, adhesion formation was evaluated. RESULTS: Firm adhesions were seen in the abrasion-no treatment group. XL reduced adhesion formation significantly compared with abrasion-no-treatment, HA, Seprafilm, and Interceed groups. Although not statistically significant, the magnitude of adhesion formation was decreased in the XL group in comparison to the PGA group. CONCLUSIONS: We suggest that the anti-adhesive effect of XL was superior to other materials in this rat model. XL application may have attenuated tissue adhesion by forming a viscous hydrogel over the injured surfaces. PMID- 17462470 TI - A novel technique for heterotopic vascularized pancreas transplantation in mice to assess ischemia reperfusion injury and graft pancreatitis. AB - BACKGROUND: Although various suture techniques for murine pancreas transplantation have been described, severe limitations have limited their widespread use. We therefore designed a surgical model for cervical heterotopic pancreas transplantation using a cuff technique. METHODS: C57BL6 mice were used as donor and recipient pairs. Recipients were rendered diabetic with streptozotocin and subsequently transplanted. The donor pancreas was isolated using a no-touch technique and then placed in the recipient's cervical region. Vascular anastomoses were completed by pulling the portal vein over the external jugular vein cuff and the donor aortic segment over the carotid cuff and fixed with an 8-0 ligature thereby facilitating a nonsuture technique. To test applicability of this model, graft microcirculation was evaluated by intravital microscopy after prolonged cold ischemia (16 h). RESULTS: The immediate success rate was >90%. Donor operation lasted 40 +/- 5 min; dissection of recipient vessels lasted 20 +/- 4 min. Revascularization time was 4 to 6 min, resulting in a total pancreas ischemia time of 33 +/- 6 min. No thromboembolic complications on the cuff side were observed. Preoperative glucose levels were 518 +/- 59 mg/dl and returned to normal by postoperative day 1 (88 +/- 13 mg/dl). Histology on postoperative days 10 and 30 showed almost normal islet cell and acinar architecture of all grafts. In groups with prolonged cold ischemia, graft microcirculation was significantly reduced and paralleled by increased inflammation, interstitial edema, hemorrhage, acinar vacuolization, and focal areas of necrosis compared with nonischemic controls. CONCLUSIONS: This new model may provide an excellent tool to further investigate the pathophysiology as well as novel therapeutic strategies of preservation, ischemia reperfusion injury, and graft pancreatitis. PMID- 17462471 TI - Littoral cell angioma mimicking pancreatic tumor. PMID- 17462472 TI - Laparoscopic paraesophageal hernia mesh repair. PMID- 17462473 TI - Emotional evaluation and skill acquisition for proficiency-based laparoscopic skills training. PMID- 17462475 TI - Bladder defense molecules, urothelial differentiation, urinary biomarkers, and interstitial cystitis. AB - It has long been recognized that interstitial cystitis (IC) is a disease of the urothelium. In this article, we review the results of published studies and present new data concerning the precise role of the bladder epithelium in IC. We discuss bladder defenses against both the penetration of urinary solutes and bacterial adherence, and we present new information about the proteoglycans that are present on the normal bladder. Previously published results and new data presented here support the conclusion that IC involves an aberrant differentiation program in the bladder urothelium that leads to altered synthesis of several proteoglycans, cell adhesion and tight junction proteins, and bacterial defense molecules such as GP51. These findings lend support to the rationale for glycosaminoglycan replacement therapy for the treatment of patients with IC. PMID- 17462476 TI - Neural upregulation in interstitial cystitis. AB - Interstitial cystitis (IC) is a syndrome of bladder hypersensitivity with symptoms of urgency, frequency, and chronic pelvic pain. Although no consensus has been reached on the underlying cause of IC, several pathophysiologic mechanisms, including epithelial dysfunction, mast cell activation, and neurogenic inflammation, have been proposed. Despite multiple different causes of urinary cystitis, the bladder's response to cystitis is limited and typical. Animal experiments have shown upregulation of proteinase-activated receptors, tryptase, beta-nerve growth factor, inducible nitric oxide synthase, nuclear transcription factor-kappaB, c-Fos, phosphodiesterase 1C, cyclic adenosine monophosphate (cAMP)-dependent protein kinase, and proenkephalin B. After the noxious stimulus has abated, downregulation of genes appears to follow. Distention of the bladder results in the release of adenosine triphosphate (ATP) from urothelial cells, which activates purinergic P2X3 receptors. Activation by ATP of P2X3-expressing afferents is a fundamental signaling factor in bladder sensation and appears to play a role in bladder reflexes. Fos proteins present in spinal cord neurons have been shown to be upregulated in animals that have undergone cyclophosphamide-induced chemical cystitis. These and other findings suggest that neural upregulation occurs both peripherally and centrally in subjects with chronic cystitis. It is unclear whether neural mechanisms and inflammation are the cause of IC or the result of other initiating events. Neural upregulation is known to play a role in the chronicity of pain, urgency, and frequency and represents an exciting area of research that may lead to additional treatments and a better understanding of IC. PMID- 17462477 TI - The mast cell in interstitial cystitis: role in pathophysiology and pathogenesis. AB - Current evidence from clinical and laboratory studies confirms that mast cells play a central role in the pathogenesis and pathophysiology of interstitial cystitis (IC). In this article, we focus on the role of the mast cell in IC and examine the ways in which mast cells and other pathophysiologic mechanisms are interrelated in this disease. Identifying the patients with IC who have mast cell proliferation and activation will enable us to address this aspect of disease pathophysiology in these individuals with targeted pharmacotherapy to inhibit mast cell activation and mediator release. PMID- 17462478 TI - Contemporary clinical presentation of interstitial cystitis. AB - For the purpose of presenting a comprehensive review of current information, the medical literature was reviewed to extract data related to the clinical presentation of interstitial cystitis (IC). The most common symptoms at diagnosis are urinary urgency, daytime frequency, dysuria, pain, and nocturia. The most common sites where pain is localized are the suprapubic, pubic, vaginal, and genital areas. Pain is commonly triggered or exacerbated by vaginal intercourse in women or ejaculation in men. The most common findings on physical examination are dysphoric mood, suprapubic tenderness, and anterior vaginal wall tenderness. A voiding diary typically shows an increased number of voids per day and reduced volumes per void. Overall, published reports in the literature indicate that the clinical presentation of IC is variable. IC is readily suggested by history, physical examination, and voiding diary findings. PMID- 17462479 TI - Prevalence of interstitial cystitis in a primary care setting. AB - In this article, we report a study that assessed the prevalence of interstitial cystitis (IC) in a primary care office using symptom-based and improved diagnosis based assessment modalities. Over the course of 1 year, all patients > or = 18 years of age who presented for a primary care office visit were administered the Pelvic Pain and Urgency/Frequency (PUF) questionnaire. Patients with potential IC as indicated by PUF score were selected for further interview and, when appropriate, a Potassium Sensitivity Test (PST) or Anesthetic Bladder Challenge (ABC). Those given the PST were queried afterward regarding the tolerability of the test. Of 3883 patients initially surveyed, 13.1% (n +/- 509) reported PUF scores suggestive of probable IC, including 17.5% (357 of 2043) of women and 8.3% (152 of 1840) of men. Overall, 4.3% (168 of 3883) of patients in this primary care population was diagnosed with IC on the basis of history, PUF score, patient interview, and results of the PST or ABC. The PST was found to be comparable to, and in most cases less painful than, several standard office-based procedures. IC is a prevalent disease in the general primary care population. The PUF questionnaire represents an easy-to-use approach for IC symptom screening, and the PST and the ABC are useful and relatively noninvasive adjuncts in the diagnosis of IC. PMID- 17462480 TI - Epidemiologic issues in interstitial cystitis. AB - As a result of variations in disease definition and diagnostic criteria for interstitial cystitis (IC), the performance of epidemiologic studies has been challenging. Initial prevalence studies used physician-confirmed diagnoses of IC; more recent studies, which have incorporated the use of patient responses to validated symptom questionnaires, indicate that the true prevalence of IC is much greater than the early studies suggested. Over the last decade, the recognized prevalence of IC has increased, and it is consistently greater among women compared with men. The most recent estimates indicate that at > or = 197 of every 100,000 women and > or = 41 of every 100,000 men in the United States are affected by IC. Because IC is substantially underdiagnosed, its actual prevalence may be much higher. Indeed, the disease may affect as many as 1 in 4 to 5 women and 1 in 20 men. PMID- 17462481 TI - The emerging presence of interstitial cystitis in gynecologic patients with chronic pelvic pain. AB - Emerging data are changing the pelvic pain paradigm for gynecologic patients. Historically, interstitial cystitis (IC) was rarely considered as a cause of chronic pelvic pain (CPP), but recent data suggest that IC is a common cause of CPP in gynecologic patients and perhaps is even the most common cause. It is important to consider the bladder as a generator of symptoms early in the evaluation of the gynecologic patient with CPP. New tools have been developed to aid the gynecologist in ruling out IC in patients with CPP, including a new IC symptom questionnaire and the Potassium Sensitivity Test (PST). By determining whether the pain is of bladder origin, the physician can more successfully treat the patient with CPP. PMID- 17462482 TI - Chronic prostatitis/chronic pelvic pain syndrome and male interstitial cystitis: enigmas and opportunities. AB - Recently, it has been recognized that interstitial cystitis (IC) in males is a more common clinical entity than was previously thought. Further, increasing evidence suggests clinical and pathogenetic similarities between male IC and chronic nonbacterial prostatitis (CP)/chronic pelvic pain syndrome. In this article, we highlight the similarities and differences between male IC and CP and review the presentation, diagnosis, and treatment of IC in men, with particular attention to those who have received an initial diagnosis of CP. PMID- 17462483 TI - Current diagnosis of interstitial cystitis: an evolving paradigm. AB - Our approaches to the diagnosis of interstitial cystitis (IC) are evolving as a result of recent advances in our knowledge of the disease. With increasing awareness of IC prevalence and presentation, clinicians are identifying cases of IC earlier in the disease process. A diagnosis of IC can now be established without applying each step of the traditional diagnostic paradigm, which was designed to identify "classic" cases of IC. In this article, we present an updated paradigm for the diagnosis of IC based on recent clinical data and consensus discussions conducted at the International Consultation on Interstitial Cystitis in Japan (ICICJ) and the National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)/Interstitial Cystitis Association (ICA) research symposium in 2003. The diagnosis is established on the basis of a thorough physical examination and the patient's history of urgency/frequency and/or pain in the absence of bacterial infection or malignancy. Use of a symptom questionnaire to capture and record the presence of all IC symptoms is helpful in establishing the diagnosis. In this evolving paradigm, all other diagnostic measures are optional. Evidence-based medicine does not require the use of either cystoscopy or urodynamics in a workup for IC. PMID- 17462484 TI - Rational approaches to the treatment of patients with interstitial cystitis. AB - Symptoms of interstitial cystitis can usually be successfully managed with heparinoid therapy to theoretically alter bladder urothelial abnormalities, and with oral medications to inhibit neural upregulation or to control mast cell dysfunction. Other forms of care ranging from intravesical therapy to endoscopic, percutaneous, or open surgery are options that may be used singly or in combination to optimize symptom reduction. PMID- 17462485 TI - Successful management of interstitial cystitis in clinical practice. AB - Primary care physicians, urologists, and gynecologists have the opportunity to detect interstitial cystitis (IC) in its early stages in symptomatic patients and provide effective treatment before the disease progresses. In this article, we present guidelines for clinical practice management and coding for reimbursement for the care of patients with IC. Important issues in the management of IC are presented, including appropriate Current Procedural Terminology (CPT) coding for office visits and procedures associated with diagnosis and treatment of the disease. Excellent IC care can be integrated into a successful clinical practice with appropriate clinical management and coding for reimbursement. PMID- 17462486 TI - The role of the urinary epithelium in the pathogenesis of interstitial cystitis/prostatitis/urethritis. AB - The urothelium plays a pivotal role as a barrier between urine and its solutes and the underlying bladder. Bladder surface mucus is a critical component of this function. The biologic activity of mucus that imparts this barrier function is generated by the highly anionic polysaccharide components (eg, glycosaminoglycans), which are extremely hydrophilic and trap water at the outer layer of the umbrella cell. This trapped water forms a barrier at the critical interface between urine and the bladder. The result is a highly impermeable urothelium that serves as a key protective barrier for the bladder interstitium. In interstitial cystitis (IC), disruption of the urothelial barrier may initiate a cascade of events in the bladder, leading to symptoms and disease. Specifically, epithelial dysfunction leads to the migration of urinary solutes, in particular, potassium, that depolarize nerves and muscles and cause tissue injury. Exogenous heparinoids can restore the barrier function of the urothelium and thus successfully treat patients with IC. Groups of patients who have been given a diagnosis of IC, chronic prostatitis, and urethritis have been shown to have IC by virtue of their shared potassium sensitivity. It would seem, therefore, that mucous deficiency may be present throughout the lower urinary tract. If one is to rename these diseases, perhaps it is best to do so in reference to a shared loss of epithelial barrier function. A name such as lower urinary dysfunctional epithelium would incorporate all of these diseases under a single pathophysiologic process. As a result of these discoveries, a new paradigm for diagnosis and treatment is emerging. PMID- 17462488 TI - Early, aggressive nutritional management for very low birth weight infants: what is the evidence? AB - The American Academy of Pediatrics Committee on Nutrition goal of providing nutrient intakes that permit the rate of postnatal growth and the composition of weight gain to approximate that of a normal fetus of the same postmenstrual age is rarely met by extremely low gestational age neonates. Therefore, postnatal growth failure or growth restriction continues to be a problem experienced by many of these infants, and they are often less than the 10th percentile of reference intrauterine curves at the time of hospital discharge. Variation in nutritional practices, especially those practices concerned with the initiation and advancement of parenteral and enteral nutrition, largely explain the difference in growth observed at different newborn intensive care units. Although limited, the evidence supports recommendations to administer early parenteral and enteral nutrition, specifically initiation of an amino acid infusion providing about 3 g protein/kg/d within hours of birth, initiation of a lipid emulsion of 0.5 to 1.0 g lipids/kg/d within 24 to 30 hours of birth, and the initiation of minimal enteral feedings within the first 5 days of life. It is important that neonatal clinicians recognize the barriers and obstacles to the implementation of these recommendations. PMID- 17462489 TI - Evidence supporting early nutritional support with parenteral amino acid infusion. AB - Postnatal growth of extremely low birth weight (ELBW) infants remains poor and does not come close to approximating rates of in utero growth. There is good evidence that early deficiencies in protein may be an important contributor to the poor growth outcomes observed in this population. Protein losses are inversely related to gestational age, and ELBW infants lose 1% to 2% of their total endogenous body protein stores each day that they receive glucose alone. It is now abundantly clear from a variety of studies that providing intravenous amino acids to sick premature infants in early postnatal life can improve protein balance and can increase protein accretion, even at low caloric intakes. Provision of approximately 1 g/kg/day of amino acids will result in a net protein balance close to zero, whereas delivery of 3 g/kg/day will accomplish protein accretion. Although data from metabolic studies, observational studies, and even a few randomized clinical trials overwhelmingly support the short-term safety and efficacy of early amino acids in reversing protein loss, there is much less known about the effects of early amino acid administration on longer-term outcomes such as growth and neurodevelopment in extremely premature infants. Based on the sum of currently available evidence presented, providing ELBW infants with 2.5 to 3.5 g/kg/day of intravenous amino acids as soon as possible after birth is a reasonable recommendation. Future studies are required to determine whether provision of 3 to 3.5 g/kg/day of amino acids is "aggressive" enough for optimal growth and neurodevelopmental outcome of ELBW infants. PMID- 17462490 TI - Dilemmas initiating enteral feedings in high risk infants: how can they be resolved? AB - In initiating enteral feedings for high-risk infants, clinicians struggle with three fundamental questions: When should enteral feedings be initiated? Should a period of trophic (minimal) feeding be provided? When feedings are advanced, how rapidly should the volume be increased? We present the findings of our systematic reviews of randomized trials addressing each of these questions. These reviews identified various limited short-term benefits of initiating feedings early, providing a period of trophic feedings, and increasing the volume at a relatively rapid rate when feedings are advanced. However, the safety and effectiveness of these approaches are unclear due to limitations in trial design, an inadequate sample size, and the problems inherent in evaluating the effects of initial feeding regimen on necrotizing enterocolitis (NEC) and neurodevelopmental outcome. We provide a detailed description of how a multicenter clinical trial might best be designed to adequately address these questions. In our view, it would be necessary to assess the effect of three feeding regimens on survival without neurodevelopmental impairment (primary outcome) among extremely low birth weight (ELBW) infants. The most daunting obstacle to resolving our current feeding dilemmas is the sample size required to assess all important outcomes. Even in the largest existing research network that achieves a high follow-up rate (the NICHD Neonatal Research Network), it is not feasible to meet conventional (frequentist) sample size requirements. Fortunately, this problem may be addressed using Bayesian methods. (For this reason and because Bayesian methods are likely to be increasingly used in neonatal trials, we provide a brief introduction to these methods.) We show that, with the sample size achievable in the Neonatal Network, Bayesian analyses are likely to provide clear and clinically useful assessments of the probability of benefit for all important clinical outcomes resulting from initial feeding regimens for ELBW infants. PMID- 17462491 TI - Enteral intake for very low birth weight infants: what should the composition be? AB - Providing optimal nutrition to satisfy the growth needs of very low birth weight infants is critical. The available preterm formulas and fortified human milk diets provide protein intakes of approximately 3.5 to 3.6 g/kg/d when volumes sufficient to provide 120 kcal/kg/d are fed to these infants. These intakes support growth and protein accretion at about or slightly greater than intrauterine rate and lead to relatively increased fat deposition. However, most very low birth infants fed these diets remain below the 10th percentile of the intrauterine growth standards at discharge. There is clear evidence that, with respect to growth, very low birth infants are likely to benefit from a higher protein intake; however, there is no clear evidence that energy intakes greater than 120 kcal/kg/d are needed. Although the upper limit of protein intake and the ideal protein:energy ratio remain controversial, there is enough evidence to support the beneficial and safe use of formulas providing protein:energy ratio of 3.2 to 3.3 g/100 kcal. PMID- 17462492 TI - Human milk and clinical outcomes in VLBW infants: how compelling is the evidence of benefit? AB - Significant benefits to infant host defense, sensory-neural development, gastrointestinal maturation, and some aspects of nutritional status are observed when premature infants are fed their mothers' own milk. A reduction in infection related morbidity in human milk-fed premature infants has been reported in nearly a dozen descriptive, and a few quasi-randomized, studies in the past 25 years. Studies on neurodevelopmental outcomes have reported significantly positive effects for human milk intake on mental and motor development, intelligence quotient, and visual acuity compared with the feeding of formula. Human milk-fed infants also have decreased rates of re-hospitalization after discharge. It is unclear how much human milk is needed to provide protection or at what postnatal age the protective effects maximize. More data are warranted to elucidate these questions. Despite the significant benefits of mothers' own milk, nutritional adequacy may be a limiting factor in the infant weighing less than 1500 g at birth. The overall nutritional needs of these infants can be supported with a nutrient supplement, or fortifier, added to the milk. PMID- 17462493 TI - Post-discharge nutrition: what does the evidence support? AB - Although there are no official recommendations for specific nutrient intakes in premature infants after hospital discharge, it is agreed that the goal should be to achieve the body composition and rate of growth of that of a normal fetus of the same postmenstrual age during the entire first year of life. A general recommendation to use the special formulas designed for preterm infants after hospital discharge in place of the formulas for term infants cannot be made from the available evidence at this time. Infants fed human milk after discharge are of the greatest concern as human milk does not in theory meet the requirements for growth in these infants. Such infants should remain on supplemental vitamins and Fe while breastfeeding, and growth as well as serum levels of phosphorus and alkaline phosphatase should be carefully monitored. The increased risk of preterm infants for obesity and the metabolic syndrome secondary to the metabolic/nutritional events early in life (programming) is likely to be small compared with the contribution of other risk factors, such as parental size, weight as an adolescent, and various lifestyle factors such as physical activity. PMID- 17462494 TI - Anthropometric and laboratory assessment of very low birth weight infants: the most helpful measurements and why. AB - Very low birth weight (VLBW; < or =1500 g), preterm infants have numerous physiological and developmental concerns, including growth and the provision of adequate nutrients to sustain growth. Growth is an important health care outcome measure for VLBW infants. Provision of energy and nutrients at levels to support growth and development is the goal of nutrition support for VLBW infants. Anthropometry and laboratory data are useful components of growth and nutrition assessment. The objectives of this paper are to describe: 1) the clinical application and interpretation of anthropometric measures of growth, and 2) the utilization and interpretation of laboratory tests of nutritional status in VLBW infants. PMID- 17462495 TI - Nutritional management of infants with short bowel syndrome. AB - The prevalence of short bowel syndrome appears to be increasing because of more aggressive surgical and medical approaches to the management of neonatal intraabdominal catastrophies. Hence, a large cohort of neonates with intestinal failure occupies neonatal intensive care units, requiring chronic total parenteral nutrition (TPN) in hopes that the residual bowel will adapt, thereby permitting weaning of TPN. Alternatively, when there is no hope for adaptation, these infants are maintained on TPN in hopes that they will grow to a size and state of general health satisfactory for either isolated intestinal transplant when liver function is preserved or combined liver-intestinal transplantation when the liver is irreparably damaged. Thus, it is imperative to provide enough parenteral nutrition to facilitate growth while minimizing TPN constituents predisposing to liver damage. Liver disease associated with intestinal failure (IFALD) seems to occur due to a variety of host factors combined with deleterious components of TPN. Host factors include an immature bile secretory mechanism, bile stasis due to fasting, and repeated septic episodes resulting in endotoxemia. Many constituents of TPN are associated with liver damage. Excessive glucose may result in fatty liver and/or hepatic fibrosis, excessive protein may lead to reduced bile flow, and phytosterols present in intravenous lipid may produce direct oxidant damage to the liver or may impede cholesterol synthesis and subsequent bile acid synthesis. Parenteral strategies employed to minimize TPN damage include reducing glucose infusion rates, reducing parenteral protein load, and reducing parenteral lipid load. Furthermore, preliminary studies suggest that fish oil-based lipid solutions may have a salutary effect on IFALD. Ultimately, provision of enteral nutrition is imperative for preventing or reversing IFALD as well as facilitating bowel adaptation. While studies of trophic hormones are ongoing, the most reliable current method to facilitate adaptation is to provide enteral nutrition. Continuous enteral feeding remains the mainstay of enteral nutrition support. PMID- 17462496 TI - Care and management of the infant of the HIV-1-infected mother. AB - Mother-to-child transmission of the human immunodeficiency virus continues to be a major global health problem. The pediatric HIV-1 epidemic is fueled by HIV-1 infection in women of childbearing age with vertical transmission in utero or at the time of birth. In resource-rich countries, the birth of an infected child is a sentinel health event signaling a chain of missed opportunities and barriers to prevention. Because the fate and ultimate HIV-infection status of the baby is inextricably linked to the infection status of the mother and her general state of well-being, we provide in this review: 1) background and state-of-the-art management guidelines for optimum maternal care; 2) strategies to minimize the risk of vertical transmission of HIV; and 3) recommendations for managing infants born to HIV-infected women. These are discussed under four case scenarios that obstetric and pediatric providers frequently encounter in their practices. PMID- 17462497 TI - Asthma and the unified airway. AB - Inflammatory processes of the upper and lower airway commonly co-exist. Patients with upper respiratory illnesses such as allergic rhinitis and acute and chronic rhinosinusitis often present to both otolaryngologists and primary care physicians for treatment of their symptoms of nasal and sinus disease. These patients often have concurrent lower respiratory illnesses such as asthma that may be contributing to their overall symptoms and quality of life. Unfortunately, asthma frequently remains undiagnosed in this population. It was the objective of this paper to examine the relationship between upper respiratory illnesses such as rhinitis and rhinosinusitis and lower respiratory illnesses such as asthma, and to provide a framework for primary care and specialty physicians to approach these illnesses as a spectrum of inflammatory disease. The present manuscript was developed by a multidisciplinary workgroup sponsored by the American Academy of Otolaryngic Allergy. Health care providers in various specialties contributed to the manuscript through preparation of written materials and through participation in a panel discussion held in August 2006. Each author was tasked with reviewing a specific content area and preparing a written summary for inclusion in this final document. Respiratory inflammation commonly affects both the upper and lower respiratory tracts, often concurrently. Physicians who are treating patients with symptoms of allergic rhinitis and rhinosinusitis must be vigilant to the presence of asthma among these patients. Appropriate diagnostic methods should be used to identify individuals with concurrent respiratory illnesses, and comprehensive treatment should be instituted to reduce symptoms and improve quality of life. PMID- 17462498 TI - Donor-recipient combinations of group A and B KIR haplotypes and HLA class I ligand affect the outcome of HLA-matched, sibling donor hematopoietic cell transplantation. AB - The influence of donor and recipient killer immunoglobulin-like receptor (KIR) genotype on the outcome of hematopoietic cell transplantation between human leukocyte antigen (HLA)-matched siblings was investigated. Transplants were divided into four groups according to the combination of group A and B KIR haplotypes in the transplant donor and recipient. Overall survival of myeloid patients varied with KIR genotype combination. Best survival was associated with the donor lacking and the recipient having group B KIR haplotypes; poorest survival was associated with the donor having and the recipient lacking group B KIR haplotypes. The latter combination was also associated with increased relapse and acute graft-versus-host disease (GVHD). However, its detrimental effects were seen only for transplants where the recipient and donor were homozygous for the C1 KIR ligand and therefore lacked the C2 ligand. Presence of the Bw4 ligand was also associated with increased acute GVHD. In contrast presence of both KIR3DL1 and its cognate Bw4 ligand was associated with decreased nonrelapse mortality. Analysis of the KIR genes individually revealed KIR2DS3 as a protective factor for chronic GVHD. The results suggest how simple assessments of KIR genotype might inform the selection of donors for hematopoietic cell transplantation. PMID- 17462499 TI - Early activation markers of human peripheral dendritic cells. AB - Two major populations of dendritic cells (DCs), myeloid and plasmacytoid, can be isolated from human peripheral blood, and are distinguished by differential expression of the cell surface markers CD11c and CD123. These two populations of DCs also are different in their expression of Toll-like receptor (TLRs), which are involved in their activation. To investigate the early events during activation of peripheral DCs, the cells were stimulated in vitro with ligands for TLR-4 (as in lipopolysaccharides [LPS]) or TLR-9 (CpG-containing oligonucleotide [CpG]). The earliest change in protein expression detected after stimulating peripheral DCs with lipopolysaccharide (LPS) or CpG was increased production of the chemokine interleukin (IL)-8. Enhanced production of IL-8 occurred already within 2 hours of stimulation in both myeloid dendritic cells (M-DCs) and plasmacytoid dendritic cells (P-DCs), and preceded expression of the well established activation marker CD40. Although both populations of DCs secreted IL 8 upon activation, the levels of IL-8 produced was several times higher within the M-DCs compared with the P-DCs population. Before activation, both subsets of DCs expressed the IL-8 receptor type B (CD128b); but after stimulation the IL-8 receptor was down-regulated in both populations of DCs. Increased expression of MHC class II molecules is generally regarded as an early activation marker of DCs. However, only the P-DCs showed a significant up-regulation of MHC class II after stimulation. The M-DC population up-regulated MHC class II without any prior activation; thus care should be taken using increased expression of MHC class II molecules as an early activation marker of peripheral M-DCs after activation in vitro. In conclusion, we propose that during activation of human DCs the production of IL-8 and loss of CD128b are the earliest signs of activation preceding both MHC class II, CD40, CD80, and CD86 expression. PMID- 17462500 TI - alpha-fetoprotein and interleukin-18 gene-modified dendritic cells effectively stimulate specific type-1 CD4- and CD8-mediated T-Cell response from hepatocellular carcinoma patients in Vitro. AB - The T-helper 1 (Th1) immune reaction is most important in dendritic cell (DC) based immunotherapy. Interleukin (IL)-18, a Th1-biasing cytokine, plays a pivotal role in inducing cytotoxic T lymphocyte (CTL) responses. In this study, we analyzed whether dendritic cells (DCs) from patients with hepatocellular carcinoma (HCC) can be transduced with the IL-18 gene and/or alpha-fetoprotein (AFP) gene, and we examined whether vaccinations using these genetically engineered DC can induce stronger therapeutic antitumor immunity. The results showed that DC transfected with AdIL-18/AFP can expressed IL-18 and AFP by reverse transcriptase-polymerase chain reaction and enzyme-linked immunoassay. Compared with those before transfection, the expressions of membrane molecules were increased dramatically. Specific T cells generated by DC transfected with AdIL-18/AFP recognized HLA-matched HepG2 cell lines specifically. Most importantly, The cytotoxic activity of CTLs against HepG2 with DC expressing AFP(AFP-DC) was significantly augmented by co-transduction with the IL-18 gene. Administration with such vaccine also significantly increased the production of interleukin-12p70 and interferon-gamma. These results indicate that a vaccination therapy using DC co-transduced with the TAA gene and IL-18 genes is effective strategy for immunotherapy in terms of the activation of DCs, CD4+ T, cells and CD8+ T cells, and may be useful in the clinical application of a cancer vaccine therapy. PMID- 17462501 TI - Correlates of CD4+ and CD8+ lymphocyte counts in high-risk immunodeficiency virus (HIV)-seronegative women enrolled in the women's interagency HIV study (WIHS). AB - Studies of human immunodeficiency virus (HIV) infection often compare values from HIV-uninfected controls, including CD4 and CD8 lymphocyte counts. Nonetheless, little is known regarding factors associated with CD4 and CD8 cell numbers in HIV uninfected individuals. To ascertain potential factors associated with differences in CD4 and CD8 cells among HIV negative women, we studied these cells in a group of 953 women, enrolled as HIV-negative comparators in the Women's Interagency HIV Study. Using standard techniques, we measured CD4 and CD8 cells obtained during study-related visits every six months through visit 20 (maximum of 9.5 years). Results were correlated with demographic and behavioral factors, and data were analyzed using a multiple linear regression approach with generalized estimating equations. At baseline, the median age was 32.4 years, body mass index (BMI) was 26.4 kg/m(2), CD4 cell count was 1010 (range 214 2705)/microL, and CD8 cell count was 542 (range 72-2448)/microL. African Americans comprised 54%, 24% were Hispanic, and 19% were Caucasian. In multivariate analysis, increasing age (p = 0.0006), increasing BMI (p = 0.001), and current smoking status (p = 0.03) were independent predictors of higher CD4 counts. Multivariate analyses of CD8 cells revealed that lower age (p = 0.001), higher BMI (p = 0.03), Hispanic race/ethnicity (p = 0.01); current smoking (p = 0.006), injection drug use (p = 0.02), and Hepatitis C infection (p = 0.01) were independent predictors of higher CD8 cell counts. Multiple demographic and behavioral factors may influence CD4 and CD8 counts in HIV negative women. These factors must be considered in future analyses comparing lymphocyte subsets in HIV positive and negative women. PMID- 17462502 TI - A novel approach to detect donor/recipent immune responses between HLA-identical pairs. AB - Renal transplant rejection and graft versus host reactions between HLA genetically-identical sibling (HLAgi) donor/recipient (D/R) pairs are thought to result from minor histocompatibility antigen (mHAg) disparities. We have compared two methods of measuring HLAgi D/R T lymphocyte responses to "matured" dendritic cells: 1.) a modified Cylex assay of CD4(+) ATP levels (MLDC-ATP) versus 2.) (3)H thymidine uptake (MLDC-(3)H). The MLDC-ATP kinetics peaked at 48 hours versus the MLDC-(3)H at 7 days, and appeared more sensitive. We tested HLAgi (normal) volunteer siblings (NLs), and D/R sibling pairs before and after renal transplantation (pre-Tx and post-Tx). The overall frequencies of positive responses in the MLDC-ATP for HLAgi NLs, pre-Tx, and post-Tx D/R pairs were 63%, 50%, and 42%, respectively. The percentage with reciprocal responses was 37.5%, 20%, and 22.22%, respectively. In one set of three HLAgi (NLs) siblings (two males and one female), there was a nongender-associated differential response. There was no MLDC correlation with class I MHC-associated mHAg (SSP) incompatibility, nor could some MLDC positive reactive pairs theoretically process the necessary HLA-class I restriction molecules for presentation of known (nanomeric) mHAg peptides. Speculatively, the MLDC reflects class II MHC restricted mHAg reactions (not yet definable), with possible effects of other polymorphic (nonhistocompatibility) immune response genes, and thereby may be a useful measurement of CD4(+) T-cell HLAgi transplantation immunity. PMID- 17462503 TI - Post transplant development of MICA and anti-HLA antibodies is associated with acute rejection episodes and renal allograft loss. AB - This study was undertaken with the primary aim of analyzing the clinical relevance of posttransplant appearance of anti-human leukocyte antigen (HLA) and major histocompatibility (MHC) class I related chain A (MICA) antibodies in response to live related donor (LRD) renal transplantation. A total of 185 consecutive post renal transplant recipient serum samples were analyzed for the detection of anti-HLA by enzyme-linked immunoabsorbent assay (ELISA) and MICA antibodies using Luminex techniques. Patients with IgG HLA class I antibodies had more acute rejection episodes compared to the negative group (67% vs. 20%, chi(2) = 7.95, p = 0.005) and also had poor graft survival (44% vs 86%, chi(2) = 6.67, p = 0.01). Similarly, patients with anti-HLA class II antibodies also had significantly lower graft survival and a higher number of rejection episodes as compared to the antibody negative group (p = 0.002 and p = 0.000, respectively). Following transplantation, 30 patients (16%) developed antibodies against any of the MICA alleles (MICA*001, MICA*002, MICA*004, MICA*008, or MIC*009). The graft survival was significantly compromised in these patients as compared to the negative group (60% vs 86%, chi(2) = 10.26, p = 0.001). Further, patients carrying both antibodies (MICA+/HLA+) were the worst affected and showed significantly poor graft survival as compared to the MICA-/HLA- group (17% vs 89%, chi(2) = 19.63, p = 0.000). Similarly, patients with only MICA antibodies or those with only HLA antibodies also had significantly lower graft survival and a higher number of acute rejection episodes (p = 0.035 and p = 0.001, respectively) as compared to the nonsensitized group. The study illustrates that posttransplant monitoring of antibodies to both MICA as well as HLA could be an important prognostic marker in renal transplant subjects. PMID- 17462504 TI - Repeated hematopoietic stem and progenitor cell mobilization without depletion of the bone marrow stem and progenitor cell pool in mice after repeated administration of recombinant murine G-CSF. AB - Administration of recombinant-human G-CSF (rhG-CSF) is highly efficient in mobilizing hematopoietic stem and progenitor cells (HSC/HPC) from the bone marrow (BM) toward the peripheral blood. This study was designed to investigate whether repeated G-CSF-induced HSC/HPC mobilization in mice could lead to a depletion of the bone marrow HSC/HPC pool with subsequent loss of mobilizing capacity. To test this hypothesis Balb/c mice were treated with a maximum of 12 repeated 5-day cycles of either 10 microg rhG-CSF/day or 0.25 microg rmG-CSF/day. Repeated administration of rhG-CSF lead to strong inhibition of HSC/HPC mobilization toward the peripheral blood and spleen after >4 cycles because of the induction of anti-rhG-CSF antibodies. In contrast, after repeated administration of rmG CSF, HSC/HPC mobilizing capacity remained intact for up to 12 cycles. The number of CFU-GM per femur did not significantly change for up to 12 cycles. We conclude that repeated administration of G-CSF does not lead to depletion of the bone marrow HSC/HPC pool. PMID- 17462505 TI - Is FCRL3 a new general autoimmunity gene? AB - Autoimmunity is a multistep pathogenic process, which arises in genetically predisposing individuals as a result of the harmful influence of environmental factors causing the breakdown of immune tolerance and induction of self-reactive immune response. Recent findings resolved common pathogenic mechanisms shared between different autoimmune diseases and suggested for the existence of genetic loci that could be involved in general autoimmunity and hence contribute to susceptibility of several autoimmune diseases. To date, several loci responsible for general autoimmunity have been identified. The Fc receptor-like 3 (FCRL3) gene is one of those loci for which a significant association with a number of autoimmune diseases such as rheumatoid arthritis (RA), autoimmune thyroid disease, and systemic lupus erythematosus (SLE) has been recently shown in Japanese. However, studies in Caucasians failed to confirm a strong association of this gene with RA and SLE and therefore made questionable the putative role of FCRL3 in general autoimmunity. In this review, we discuss whether the FCRL3 gene is a newly discovered gene contributing to shared susceptibility between autoimmune diseases. PMID- 17462506 TI - Molecular scanning of interleukin-21 gene and genetic susceptibility to type 1 diabetes. AB - A recent study in the nonobese diabetic (NOD) mouse demonstrated the involvement of interleukin (IL)-21 in the pathogenesis of type 1 diabetes. A strong susceptibility locus, Idd3, has also been mapped to the interval containing the murine gene for IL-21 (Il21), making Il21 and the human orthologue IL21 a functional and positional candidate gene for type 1 diabetes. To investigate the contribution of the human genes for IL-21 and its receptor (IL-21R) to susceptibility to type 1 diabetes, we re-sequenced IL21 to identify novel sequence variants, searched for informative variants of IL21R, and studied the association of these variants with the disease. Two polymorphisms, a single nucleotide polymorphism (SNP) and a mononucleotide repeat polymorphism, were identified for IL21, and an allele of the mononucleotide repeat polymorphism was positively associated with the disease. Two novel microsatellite polymorphisms of IL21R were identified, one of which was associated with the disease. Scoring of individuals according to the status of these alleles showed a significant trend for high scores for susceptibility in diabetes patients, suggesting the contribution of IL21 and IL21R to disease susceptibility in an additive manner. These data suggest a contribution of IL21 and IL21R to genetic susceptibility to type 1 diabetes and possible involvement of IL-21 and its receptor system in the disease pathogenesis. PMID- 17462507 TI - Common and well-documented HLA alleles: report of the Ad-Hoc committee of the american society for histocompatiblity and immunogenetics. AB - In histocompatibility testing some genotype ambiguities are almost always resolved into the genotype with the most common alleles. To achieve unambiguous assignments additional unwieldy tests are performed. The American Society for Histocompatibility and Immunogenetics formed a committee to define what human leukocyte antigen (HLA) genotypes do not need to be resolved in external proficiency testing. The tasks included detailed analysis of large datasets of high-resolution typing and thorough review of the pertinent scientific literature. Strict criteria were used to create a catalogue of common and well documented (CWD) alleles. In total, 130, 245, 81, and 143 of the highly polymorphic HLA-A, -B, -C, and DRB1 loci fell into the CWD category; these represent 27%-30% of all alleles recognized. For the loci DRB3/4/5, DQA1, DQB1, and DPB1, a total of 29, 16, 26, and 52 CWD alleles were identified. A recommendation indicated that an acceptable report should only include one possible genotype; multiple genotypes can only be reported if only one of these includes two alleles of the CWD group. Exceptions in which resolution is not necessary are ambiguities involving functional alleles with identical sequences in the antigen recognition site. The criteria were established for proficiency testing, which could be a valuable tool when making clinical histocompatibility decisions. PMID- 17462508 TI - Identification of a new HLA-B allele (B*1576) by haplotype specific extraction. AB - Routine typing of a potential bone marrow donor by sequence-specific oligonucleotide probes (SSOP) and sequence based typing (SBT) produced inconclusive subtyping results, suggesting a new allele. A magnetic bead-based method, haplotype specific extraction (HSE), was used to separate the diploid sample into its haploid components. The sample was then re-typed using standard SBT, revealing a new human leukocyte antigen (HLA) allele, since named B*1576. HSE used in conjunction with standard SBT is a convenient and simple tool for resolving ambiguous and novel allele combinations without the need for amplification or subcloning. PMID- 17462509 TI - The genetic influence of the nonclassical MHC molecule HLA-G on multiple sclerosis. AB - Human leukocyte antigen (HLA)-G is a nonclassical major histocompatibility complex (MHC) molecule located at MHC complex at chromosome 6 and chiefly attributed immunoregulatory and tolerogenic functions. HLA-G is upregulated at sites of inflammation in multiple sclerosis (MS) and assumed to counterbalance immune responses. Different functionally relevant genetic variants of HLA-G have been described and shown to be statistically associated with human diseases such as fetal loss or sarcoidosis. We investigated the influence of three different variations in the HLA-G gene for disease susceptibility and course of MS (n = 698): (1) The -725 C/G exchange in the HLA-G promoter region, (2) HLA-G*0105N, a deletion that results in an irregular stopcodon in exon 3, and (3) a 14 bp insertion / deletion in the untranslated exon 8. None of these variations significantly influenced the susceptibility to multiple sclerosis. No association was seen with the age of onset of disease, disease severity or disease course. Although HLA-G is assumed to play an important role in the immunoregulatory processes of MS, our results do not support a role of genetic factors influencing disease susceptibility of the disease course. PMID- 17462510 TI - High frequency of the TCRBV20S1 null allele in the Sardinian population. AB - Single nucleotide polymorphisms (SNPs) in the T-cell receptor (TCR) gene segments might play a role in shaping the TCR repertoire. Three polymorphisms have been described for the TCRBV20S1 gene segment, one of which is responsible for a nucleotide substitution at position 524, resulting in the introduction of a stop codon. Individuals homozygous for this inactivating polymorphism ("null allele") are unable to express TCRBV20 gene products. Using DNA restriction digestion analysis, we investigated the frequency of this polymorphism in 111 healthy Sardinian subjects. Inhabitants of the Mediterranean island of Sardinia are considered to represent a genetically isolated population. Our analyses revealed an incidence of 19.8% of homozygosity for the null allele, corresponding to an allele frequency of 0.45. Such an incidence, significantly higher than the one detected in 83 non-Sardinian Caucasians (6%), is the most elevated so far reported in the literature. BV20 is a single member subfamily and the null allele produces a gap in the potential TCR repertoire. Therefore, it is possible that an undetermined selective pressure could have played a role in determining the high frequency of this inactivating polymorphism in Sardinians. Alternatively, this finding could be related to a founder effect in this ancient island population. PMID- 17462511 TI - HLA haplotypes in Singapore: a study of mothers and their cord blood units. AB - During 2005, a total of 174 cord blood units with their paired maternal samples from the Singapore Cord Blood Bank were typed for HLA-A, -B, -C at intermediate resolution and DRB1 at allelic resolution. Analysis of allele segregation in mother and child assigned 185 different four locus (HLA-A, -B, -C, -DRB1) haplotypes in Chinese, 66 in Malays, and 34 in Asian Indians. Very few four locus haplotypes were shared among population groups. To evaluate the frequencies of four locus haplotypes, the Expectation Maximization algorithm was used with HLA assignments from 536 unrelated Chinese volunteers from the Singapore Bone Marrow Donor Program registry. The paired maternal and cord blood study identified 75 different B-C associations in Chinese, 52 in Malays, and 24 in Asian Indians. Common B-C associations may be shared among population groups; for example, B*4001g-Cw*0702g was common in Chinese and Malays, whereas B*1502g-Cw*0801g and B*3501g-Cw*0401g were found in all three groups. The high diversity of four locus haplotypes originates from multiple combinations of both HLA-A and -DRB1 alleles with each B-C haplotype. PMID- 17462512 TI - HLA class I polymorphism in Mongolian and Hui ethnic groups from Northern China. AB - HLA-A, -B, and -C alleles were genotyped by sequencing-based typing (SBT) in 102 unrelated ethnic Mongolian individuals living in Inner Mongolia and 110 Hui individuals inhabiting the Qihai plateau in Northern China. In all, 28 HLA-A, 49 HLA-B, and 27 HLA-C alleles in Mongolians and 29 HLA-A, 41 HLA-B, and 27 HLA-C alleles in Hui were detected in this study. A*24G1, A*110101/1121N and A*02G1 are the three most frequent HLA-A alleles both in Mongolians and Hui. At the HLA-B locus, only B*51G1 was found with a frequency of more than 10% in Hui. Cw*070201G1 is the most common HLA-C allele both in Mongolian and Hui. The most frequent HLA-A:C:B, HLA-A:C, and HLA-C:B haplotypes are A*330301-Cw*030201/030202 B*5801, A*330301-Cw*030201/030202, and Cw*030201/030202-B*5801 in Mongolian and A*0207/0215N-Cw*010201/010202-B*4601, A*02G1-Cw*070201G1, and Cw*010201/010202 B*4601 in Hui, respectively. The genetic distance (GD) estimated according to HLA A, -B, and -C allele frequency indicates that Mongolian and Hui have the closest relationship, and both are closer to Northern Han rather than Southern Han, suggesting that the two ethnicities might have been subjected to intensive gene exchange with Northern Han in history. The dendrogram based on the GD measurements further demonstrates that Mongolian and Hui cluster as a branch with Northern Han Chinese and Northeast Asians. Our results may lead to better understanding of the origins and relationships of Chinese ethnic groups and provide the genetic background for disease association studies. PMID- 17462513 TI - Comparison distribution of HLA-B alleles in mexican patients with takayasu arteritis and tuberculosis. AB - A possible relationship between Takayasu arteritis (TA) and Tuberculosis (Tb) has been suggested. Both diseases present similar chronic inflammatory lesions and occasionally granulomas on the arterial walls. The genetic relationship between these two diseases has not been explored before, however, both diseases have been associated with human leukocyte antigen (HLA) alleles. Therefore, the aim of the present study was to analyze the distribution of HLA-B alleles in TA (n = 40) and Tb (n = 34) patients and healthy controls (72 exposed and 99 nonexposed). HLA-B alleles were determined by reverse dot blot. The statistical methods used included the Chi(2), and odds ratio (OR) with 95% confidence intervals. In spite of the loose clinical relationship between TA and Tb, we did not detected any genetic relationship between them when the HLA-B alleles were analyzed in these groups of patients. On the contrary, we detected distinct specific HLA-B alleles for each disease. TA was characterized by HLA-B39, -B44, and -B52, pulmonary Tb by HL-B35 and extrapulmonary Tb by HLA-B39 and -B40. This preliminary study suggests a difference in the distribution of HLA-B alleles in patients with TA and Tb. PMID- 17462514 TI - Polymorphism in the CCR5 gene promoter and HIV-1 infection in North Indians. AB - The clinical course and outcome of human immunodeficiency virus-1 (HIV-1) infection are highly variable among individuals. CCR5 is the primary coreceptor that mediates entry of HIV-1 (R5) into permissive host cells. In this study, five SNPs (59029G/A, 59353T/C, 59356C/T, 59402A/G, and 59653C/T) in the promoter region and a deletion of 32 bp (Delta32) in the CCR5 gene were evaluated in 180 chronically HIV-1-infected North Indians. The study showed the following: (1) the protective CCR5 Delta32 allele was absent; (2) the frequency of CCR5*59402A allele in the HIV-infected people (66.4%) was higher than in healthy subjects (57.1%, p = 0.027) and in the CDC stage C patients (76%) versus stages A and B patients together (60%; p = 0.002); (3) homozygous CCR5*59402 AA genotype was significantly increased in the seropositive subjects (46.1%) compared with healthy control subjects (30.2%; p = 0.008) and in the CDC stage C patients (59.2%) compared with stage A and B subjects (37.6%, p = 0.007); and (4) an increased frequency of homozygous ACCAC haplotype was present in the seropositive stage C patients (32.4%) versus 15.6% in patients in stages A plus B (p = 0.013). These observations suggest an association of CCR5*59402A with increased likelihood of acquisition of HIV-1 and development of AIDS in the Asian Indian population. Further studies are required to confirm these findings and understand the effect of CCR5 polymorphisms on the outcome of HIV-1 infection. PMID- 17462516 TI - Abdominal pain. AB - Evaluation of the elderly patient with acute abdominal pain is sometimes difficult. Various factors can obscure the presentation, delaying or preventing the correct diagnosis and leading to adverse patient outcomes. Clinicians must consider multiple diagnoses, especially those life-threatening conditions that require timely intervention to limit morbidity and mortality. This article reviews abdominal pain in the elderly, discusses the clinical approach, and highlights key diagnostic considerations. PMID- 17462517 TI - Back pain in the elderly. AB - Back pain in the elderly is a complex chief complaint, because numerous life threatening or disabling conditions may present with signs and symptoms similar to those of more benign disease processes. An understanding of the limitations of physical examination and history distinguishing dangerous from benign causes is essential to avoid misdiagnosis and patient morbidity or mortality. This article describes the presentation and evaluation of dangerous causes of back pain, including vascular catastrophes, spinal cord compression syndromes, and infectious diseases. Less emergent musculoskeletal injuries are discussed briefly. PMID- 17462518 TI - Headache in the elderly. AB - Headache in an elderly patient can be a sign of serious, potentially life threatening disorders. All patients require a full assessment, including a complete neurologic examination. Particular emphasis should be placed on excluding subarachnoid hemorrhage, subdural hematoma, giant cell arteritis, intracranial neoplasm, cerebrovascular accident, acute-angle-closure glaucoma, and infectious etiologies such as meningitis and encephalitis. Once life threatening disorders are excluded, the geriatrician can focus on more benign etiologies such as migraine, tension headache, and medication withdrawal. Treatment depends on the underlying etiology. This article discusses headaches that require emergent treatment and then describes more benign etiologies of headaches. PMID- 17462519 TI - Evaluation of the acutely dyspneic elderly patient. AB - Dyspnea is among the most frequent complaints in the elderly. The prevalence of comorbid medical conditions and the physiologic changes of aging present significant challenges in determining the cause. The initial approach to the elderly dyspneic patient mandates consideration of a broad range of diagnoses. Failure to diagnose life-threatening medical conditions presenting with dyspnea such as pulmonary embolus, acute coronary syndromes, congestive heart failure, asthma, obstructive pulmonary disease, pneumothorax, and pneumonia can lead significant morbidity and mortality. This article focuses on the rapid assessment and approach to the acutely dyspneic elderly patient. PMID- 17462520 TI - Evaluation of the elderly patient with acute chest pain. AB - Chest pain is one of the most common and serious chief complaints. Geriatricians must be well versed in evaluating elderly patients who have chest pain. This article discusses the initial diagnostic evaluation of elderly patients with chest pain. Specific emphasis is placed on identifying acute coronary syndromes, aortic dissection, pulmonary embolism, and pericarditis with cardiac tamponade. By understanding the different presentations of these potentially life threatening emergencies, the geriatrician will be better prepared to distinguish them from less dire conditions and to initiate prompt treatment. PMID- 17462521 TI - Syncope in the elderly. AB - Syncope is common and costly especially in the elderly. Presentation and prevalence may be different compared with the young. Etiologies can be difficult to confirm but it is not a hopeless morass. The history, physical exam, and ECG have the greatest utility. Additional studies should be used sparingly and based on the initial data. Successful response to treatment is difficult to predict. PMID- 17462522 TI - Severe hypertension in the geriatric patient-is it an emergency or not? AB - Hypertension is a medical condition commonly seen in the outpatient setting. Primary care providers should be aware that asymptomatic hypertension, despite the degree of elevation, is rarely an emergency. Based on consensus guidelines from the Joint National Committee on Prevention, Detection, Evaluation, and Treatment of High Blood Pressure and the lack of any evidence showing harm, extreme blood pressure elevations do not need acute treatment. This article provides evidence for the argument that hypertension is rarely an emergency at all; even patients who have exceedingly high blood pressure can be treated as outpatients. PMID- 17462523 TI - Polypharmacy and the geriatric patient. AB - The older adult population is increasing and with it, the risk of polypharmacy. Multiple physicians treating one patient, increasing comorbidities, and an increase in the variety of drugs available contribute to the adverse effects of polypharmacy on the elderly patient. Application of Beers criteria, appropriate therapeutic drug monitoring, and careful, periodic review of the patient's medication list will assist with preventing the sometimes lethal complications of polypharmacy. PMID- 17462524 TI - Psychiatric emergencies in the geriatric population. AB - Failure to recognize psychiatric conditions in any age group particularly in the elderly could lead to a very complicated clinical picture and may lead to increased health expenditures. Psychiatric emergencies can be secondary to numerous factors: geriatric depression and suicide, behavioral disturbance secondary to underlying organic conditions, substance abuse, elder abuse, and medication-induced adverse events. This article discusses mainly geriatric depression, elder abuse, and conditions emanating from iatrogenic causes. PMID- 17462525 TI - Evaluation and management of transient ischemic attacks. AB - The risk for disabling stroke is greatest in the period immediately following a transient ischemic attack (TIA), thus, TIA is a medical emergency. A universities medical center's emergency department-based TIA evaluation and management program is presented as an example of the pragmatic enactment of current guidelines for TIA evaluation and management. A discussion of modifications that can be made for an outpatient setting evaluation and a review of the literature follow. PMID- 17462526 TI - Venous thromboembolic disease in the elderly patient: atypical, subtle, and enigmatic. AB - Venous thromboembolic disease (VTE) is a common medical condition in the geriatric population. Risk factors include congestive heart failure, cancer, orthopedic conditions such as hip fractures, and other debilitating diseases. Clinical presentations of patients who have VTE may be subtle and atypical. Traditional vital sign abnormalities found in patients who have VTE may be absent in the older patient, and presentations such as isolated syncope without chest pain or dyspnea are common. This article provides information that will allow primary care providers to increase their rate of detection of VTE, particularly pulmonary embolism. By having an appreciation for the subtle and atypical presentations of VTE, they will be in a position to significantly lower morbidity and mortality in older patients. PMID- 17462527 TI - Acute coronary syndromes in the elderly. AB - Elderly patients with acute coronary syndromes suffer higher mortality and more morbidity than their younger counterparts because they have higher prevalence of cardiac risk factors and have lived with those risk factors longer, physiologic effects of aging lead to sicker patients at presentation and impaired healing processes, they are more sensitive to some of the side effects of some of our higher risk therapies, and they are less likely to receive therapies that have been proven to improve outcome. Close attention must be paid to risk assessment. We must continue to strive to match risky therapies to high-risk patients, those more likely to derive benefit. Age remains one of the most powerful cardiac risk factors of all. PMID- 17462528 TI - Infections in the elderly. AB - Infectious diseases remain a significant cause of morbidity and mortality in the growing number of adults over the age of 65 years in the United States. Declining immunity coupled with aging anatomy and physiology set the stage for increased vulnerability to infections and the development of atypical presentations in the elderly. Pneumonia, urinary tract infection, and skin and soft tissue infections are illnesses commonly encountered in the care of this unique population. This article explores the etiology, diagnosis, and constantly evolving treatment of these conditions in the context of the elderly patient. PMID- 17462529 TI - [Report on the 13th Anesthesia-Resuscitation journal club in ORL]. PMID- 17462530 TI - [Report on the 20th reunion of the Anesthesia-Resuscitation Club in ORL]. PMID- 17462531 TI - Nitric oxide, mitochondrial hyperpolarization, and T cell activation. AB - T lymphocyte activation is associated with nitric oxide (NO) production, which plays an essential role in multiple T cell functions. NO acts as a messenger, activating soluble guanyl cyclase and participating in the transduction signaling pathways involving cyclic GMP. NO modulates mitochondrial events that are involved in apoptosis and regulates mitochondrial membrane potential and mitochondrial biogenesis in many cell types, including lymphocytes. Mitochondrial hyperpolarization (MHP), an early and reversible event during both activation and apoptosis of Tlymphocytes, is regulated by NO. Here, we discuss recent evidence that NO-induced MHP represents a molecular switch in multiple T cell signaling pathways. Overproduction of NO in systemic lupus erythematosus induces mitochondrial biogenesis and alters Ca(2+) signaling. Thus, whereas NO plays a physiological role in lymphocyte cell signaling, its overproduction may disturb normal T cell function, contributing to the pathogenesis of autoimmunity. PMID- 17462533 TI - Insulin-like growth factor-I (IGF-I) induces epidermal growth factor receptor transactivation and cell proliferation through reactive oxygen species. AB - Insulin-like growth factor-I (IGF-I) plays an important role in proliferation of vascular smooth muscle cells (VSMCs). However, the mechanism that IGF-I induces VSMCs proliferation is not completely understood. In this study, we determined (a) whether and how IGF-I induces transactivation of epidermal growth factor receptor (EGFR) in primary rat aortic VSMCs, (b) the contribution of EGFR to IGF I-stimulated activation of extracellular signal-regulated kinase (ERK) and cell proliferation, and (c) the role of reactive oxygen species (ROS) in the cellular function. We showed that IGF-I induced phosphorylation of EGFR and ERK1/2 in VSMCs. AG1478, an EGFR inhibitor, inhibited IGF-I-induced phoshorylation of EGFR and ERK1/2. IGF-I stimulated ROS production and Src activation. Antioxidants inhibited IGF-I-induced ROS generation and activation of EGFR, ERK, and Src. Src kinase inhibitor PP1 and Src siRNA blocked IGF-I-induced activation of EGFR and ERK1/2. Inhibition of IGF-I-stimulated EGFR activation inhibited IGF-I-induced VSMC proliferation. These results suggest that (1) IGF-I induces EGFR activation through production of ROS and ROS-mediated Src activation in VSMCs, and (2) EGFR transactivation is required for IGF-I-induced VSMC proliferation. PMID- 17462532 TI - The chemistry and biology of nitroxide compounds. AB - Cyclic nitroxides are a diverse group range of stable free radicals that have unique antioxidant properties. Because of their ability to interact with free radicals, they have been used for many years as biophysical tools. During the past 15-20 years, however, many interesting biochemical interactions have been discovered and harnessed for therapeutic applications. Biologically relevant effects of nitroxides have been described, including their ability to degrade superoxide and peroxide, inhibit Fenton reactions, and undergo radical-radical recombination. Cellular studies defined the activity of nitroxides in vitro. By modifying oxidative stress and altering the redox status of tissues, nitroxides have been found to interact with and alter many metabolic processes. These interactions can be exploited for therapeutic and research use, including protection against ionizing radiation, as probes in functional magnetic resonance imaging, cancer prevention and treatment, control of hypertension and weight, and protection from damage resulting from ischemia/reperfusion injury. Although much remains to be done, many applications have been well studied and some are currently being tested in clinical trials. The therapeutic and research uses of nitroxide compounds are reviewed here with a focus on the progress from initial development to modern trials. PMID- 17462534 TI - 4-Hydroxynonenal and PPARgamma ligands affect proliferation, differentiation, and apoptosis in colon cancer cells. AB - PPARgamma ligands inhibit growth and induce apoptosis of various cancer cells. 4 Hydroxynonenal (HNE), a product of lipid peroxidation, inhibits proliferation and induces differentiation or apoptosis in neoplastic cells. The aim of this work was to investigate the effects of PPARgamma ligands (rosiglitazone and 15-deoxy prostaglandin J2 (15d-PGJ2)) and HNE, alone or in association, on proliferation, apoptosis, differentiation, and growth-related and apoptosis-related gene expression in colon cancer cells (CaCo-2 cells). PPARgamma ligands inhibited cell proliferation (IC50 was 37.47+/-6.6 microM, for 15d-PGJ2, and 170.34+/-20 microM for rosiglitazone). HNE (1 microM) inhibited cell growth by 70%. Apoptosis was induced by 15d-PGJ2 and HNE and, to a minor extent, rosiglitazone. Differentiation was induced by rosiglitazone and by 15d-PGJ2, but not by HNE. PPARgamma ligands inhibited c-myc expression. HNE induced a transitory increase in c-myc expression and a subsequent down-regulation. HNE induced p21 expression, whereas PPARgamma ligands did not. Expression of the bax gene was increased by HNE and 15d-PGJ2, but not by rosiglitazone. No synergism or antagonism was found between HNE and PPARgamma ligands. Both apoptosis and differentiation induction may be responsible for the inhibition of proliferation by PPARgamma ligands; apoptosis and c-myc and p21 expression seem to be involved in the inhibition of proliferation by HNE. PMID- 17462535 TI - Nox1-based NADPH oxidase-derived superoxide is required for VSMC activation by advanced glycation end-products. AB - Vascular diseases are important clinical complications of diabetes. Advanced glycation end-products (AGE) are mediators of vascular dysfunction, but their effects on vascular smooth muscle cell (VSMC) ROS production are unclear. We studied the source and downstream targets of AGE-mediated ROS and reactive nitrogen species production in these cells. Significant increases in superoxide production in AGE-treated VSMC were measured using lucigenin (7650+/-433 vs 4485+/-424 LU/10(6) cells, p<0.001) or coelenterazine (277,907+/-71,295 vs 120,456+/-4140 LU/10(6) cells, p<0.05) and confirmed by ESR spectroscopy. These signals were blocked by the flavin-containing oxidase inhibitor diphenylene iodonium (DPI). AGE-stimulated NF-kappaB activity was abolished by DPI and the superoxide scavenger MnTBAP. AGE differentially regulated VSMC NADPH oxidase catalytic subunits, stimulating the transcription of Nox1 (201+/-12.7%, p<0.0001), while having no effect on Nox4. AGE also increased 3-nitrotyrosine formation, which was inhibited by MnTBAP, DPI, or the NOS inhibitor L-NAME. Regarding the source of NO, AGE stimulated inducible nitric oxide synthase mRNA (1 vs 9.7+/-3.0, p=0.046), which was abolished by a NF-kappaB inhibitor, SOD, catalase, or siRNA against Nox1. This study establishes that AGE activate iNOS in VSMC through a ROS-sensitive, NF-kappaB-dependent mechanism involving ROS generation by a Nox1-based oxidase. PMID- 17462536 TI - Nickel(II)-catalysed oxidative guanine and DNA damage beyond 8-oxoguanine. AB - Oxidative DNA damage is one of the most important and most studied mechanisms of disease. It has been associated with a range of terminal diseases such as cancer, heart disease, hepatitis, and HIV, as well as with a variety of everyday ailments. There are various mechanisms by which this type of DNA damage can be initiated, through radiation and chemical oxidation, among others; however, these mechanisms have yet to be fully elucidated. A HPLC-UV-EC study of the oxidation of DNA mediated by nickel(II) obtained results that show an erratic, almost oscillatory formation of 8-oxoguanine (8-oxoG) from free guanine and from guanine in DNA. Sporadic 8-oxoG concentrations were also observed when 8-oxoG alone was subjected to these conditions. A HPLC-MS/MS study showed the formation of oxidised-guanidinohydantoin (oxGH) from free guanine at pH 11, and the formation of guanidinohydantoin (GH) from DNA at pH 5.5. PMID- 17462537 TI - Action of Nrf2 and Keap1 in ARE-mediated NQO1 expression by quercetin. AB - Polyphenols are characterized by the presence of more than one phenolic group and are widely distributed in many fruits and vegetables. They possess antioxidant properties and interact with cellular defense systems through the antioxidant responsive element/electrophile-responsive element (ARE/EpRE) although the precise mechanism by which polyphenols influence transcription factor complexes to target ARE is poorly understood. In the present study, we chose a typical polyphenol, quercetin, to investigate the mechanism in human HepG2 cells. Quercetin enhanced the ARE binding activity and Nrf2-mediated transcription activity. Molecular evidence revealed that quercetin not only up-regulated the expression of Nrf2 mRNA and protein, but also stabilized Nrf2 protein by inhibiting the ubiquitination and proteasomal turnover of Nrf2. At the same time, quercetin markedly reduced the level of Keap1 protein in posttranslational levels through the formation of modified Keap1 protein, rather than 26S proteasome dependent degradation mechanisms, without affecting the dissociation of Keap1 Nrf2. Silencing Keap1 using Keap1 siRNA significantly increased the Nrf2 dependent ARE activity, whereas silencing Nrf2 using Nrf2 siRNA markedly reduced the ARE activity under both baseline and quercetin-induced conditions. Thus, we conclude that the pathway of quercetin-induced ARE activity involves up regulation of Nrf2 through the regulation of both transcription and posttranscription sites and repression of Keap1 by affecting the posttranscription site, revealing some substantial differences between oxidative inducers. Thus, the findings provide an insight into the mechanisms underlying polyphenolic compounds in cytoprotection and cancer chemoprevention. PMID- 17462538 TI - Identification of JWA as a novel functional gene responsive to environmental oxidative stress induced by benzo[a]pyrene and hydrogen peroxide. AB - Oxidative stress has been implicated as one of the primary mechanisms inducing DNA damage and believed to mediate aging and progression of numerous age-related diseases, including cancer. JWA, a gene previously described to mediate differentiation of leukemic cells, is also involved in cellular responses to environmental exposures linked to heat shock and chemical-mediated oxidative stresses. However, the precise pathways and mechanisms underlying these phenomena remain to be resolved. Our studies demonstrated that H(2)O(2) is the primary oxidative product responsible for benzo[a]pyrene (B[a]P)-induced JWA expression, and knockdown of JWA elevates H(2)O(2) (100 microM)- and B[a]P (100 microM) induced DNA damage. In oxidative stress cell culture models, JWA was upregulated. JWA expression regulated a parallel rise in the base excision repair protein XRCC1 but a reduction in PARP1 in response to H(2)O(2)-induced DNA damage. Furthermore, we found that both H(2)O(2) and B[a]P exposure activated nuclear transcription factor I (NFI) in NIH-3T3 cells, which specifically bound to the CCAAT element in the JWA proximal promoter (-58/-28 bp) and thereby induced JWA expression. Consistently siRNA mediated a knockdown of NFI, which prevented JWA induction. These findings indicate that JWA may serve as a novel environmental stress sensor to protect cells against reactive oxygen species-associated DNA damage. PMID- 17462539 TI - Signaling pathways involved in antioxidant control of glioma cell proliferation. AB - Tumor cells are able to survive and proliferate despite the higher-than-average level of reactive oxygen species (ROS) they exhibit. This is generally taken as a clue as to the implications of ROS in cell proliferation. In fact many mitogenic intracellular signaling pathways could be redox regulated, more particularly those involving tyrosine kinase receptors (RTK). In the present work we use N acetylcysteine (NAC)-a well-known antioxidant molecule-to study the implications of cellular redox state on rat C6 glioma cell proliferation. NAC is shown to decrease glioma cell proliferation, inducing a cell cycle arrest in the G(0)/G(1) phase and markedly up-regulating p21 expression. A rapid, and glutathione independent, decrease in intracellular oxidants was observed as well. NAC also lowers Akt activity, extracellular signal-regulated kinase 1/2, and the redox sensitive transcription factor NF-kappaB, all of which are ROS related and seem to be in close connection with cell proliferation. NAC effects apparently relate to protein kinase C (PKC) activity because 100 nM TPA-a PKC activator-induces a partial blockage of the NAC antiproliferative effect. Bringing our results together, it seems that intracellular reduction of oxidants in C6 glioma cells can induce inhibition of cell proliferation by modulating RTK-related intracellular signaling pathways. PMID- 17462540 TI - Coenzyme Q and protein/lipid oxidation in a BSE-infected transgenic mouse model. AB - Oxidative stress and antioxidants play an important role in neurodegenerative diseases. However, the exact participation of antioxidants in the evolution of prion diseases is still largely unknown. The aim of this study was to assess brain levels of coenzyme Q (CoQ), an endogenous lipophilic antioxidant, and the antioxidant/pro-oxidant status by determining oxidative damage to proteins and lipids after intracerebral bovine spongiform encephalopathy (BSE) infection of transgenic mice expressing bovine prion protein (PrP). Our results indicate that, whereas the ratio between the two CoQ homologues present in mice (CoQ(9) and CoQ(10)) is not altered by prion infection during the course of the disease, significant increases in total CoQ(9) and CoQ(10) were observed in BSE-infected mice 150 days after inoculation. This time point coincided with the first manifestation of PrP(Sc) deposition in nervous tissue. In addition, CoQ(9) and CoQ(10) levels, neuropathological alterations, and PrP(Sc) deposition in nervous tissues underwent further increases as the illness progressed. Lipid and protein oxidation were observed only at the final stage of the disease after clinical signs had appeared. These findings indicate upregulation of CoQ(9)- and CoQ(10) dependent antioxidant systems in response to the increased oxidative stress induced by prion infection in nervous tissue. However, the induction of these endogenous antioxidant systems seems to be insufficient to prevent the development of the illness. PMID- 17462541 TI - A reduction in alcohol consumption is associated with reduced plasma F2 isoprostanes and urinary 20-HETE excretion in men. AB - There is considerable evidence that chronic moderate-to-high alcohol consumption increases blood pressure. The mechanisms by which this occurs are not clear. Alcohol consumption can induce oxidative stress and cytochrome P450 (CYP450) isoforms that are associated with oxidative stress and may influence vascular tone. To study the role of such mechanisms we examined whether reducing alcohol intake in moderate-to-heavy drinkers (40-110 g/day) resulted in changes in urinary excretion of 20-HETE, a CYP450 metabolite of arachidonic acid, and plasma and urinary F(2)-isoprostanes as markers of lipid peroxidation. After a 4-week run-in period during which healthy men maintained their usual drinking pattern they were randomized to a two-way crossover intervention study. In each of the 4 week treatment periods subjects either substituted their usual alcohol intake with a 0.9% alcohol beer or maintained their usual alcohol intake. Plasma and urinary F(2)-isoprostanes and urinary 20-HETE were measured by gas chromatography mass spectrometry, and serum gamma-glutamyl transpeptidase (gamma-GT) was measured as a biomarker of alcohol consumption, at the end of each study period. Sixteen healthy men age 51.0+/-2.7 years and with a BMI of 26.4+/-0.61 kg/m(2) completed the study. The reductions in alcohol intake (72.4+/-5.0 vs 7.9+/-1.6 g/day, p<0.001) and serum gamma-GT (geometric mean 24.4 U/L (95% CI 19.7, 30.2) vs 18.6 U/L (95% CI 15.5, 22.2, p<0.01) were accompanied by a significant fall in blood pressure as well as urinary 20-HETE excretion (158+/-23 vs 109+/-19 pmol/mmol creatinine, p<0.001) and plasma F(2)-isoprostanes (3438+/-158 vs 2929+/ 145 pmol/L, p=0.01). A substantial reduction in alcohol consumption in healthy men lowered plasma F(2)-isoprostanes and urinary 20-HETE. Increased oxidative stress and 20-HETE production may be linked, at least in part, to the pathogenesis of alcohol-related hypertension. PMID- 17462542 TI - Four thiol peroxidases contain a conserved GCT catalytic motif and act as a versatile array of lipid peroxidases in Anabaena sp. PCC7120. AB - The Anabaena sp. (ANASP) genome contains seven open reading frames with homology to thiol peroxidase (TPx), also known as peroxiredoxin (Prx). Based on sequence similarities among putative TPx's derived from various cyanobacteria genomes, we designated the seven putative TPx members as VCP, VCT, TCS, and GCT clusters according to the sequence of their conserved catalytic motif. The GCT cluster consists of four members, named GCT1, GCT2, GCT3, and GCT4. The ANASP GCT-TPx genes were recombinantly expressed in Escherichia coli. The purified proteins were characterized with an emphasis on the ability to destroy various peroxides, the electron donor, and the conserved cysteine structure as a catalytic intermediate. All GCT members, as an atypical 2-Cys TPx family, exerted the highest peroxidase activity toward a lipid hydroperoxide using an electron from thioredoxin. Periplasmic protein analysis revealed that GCT2 and GCT4 are distributed in the cytoplasm, whereas GCT1 and GCT3, homologues of E. coli bacterioferritin comigratory protein/plant PrxQ, are localized in the periplasmic space. Immunoblots of the heterocystic proteins showed that the level of GCT2 in the heterocyst is comparable to that in the vegetative cell, whereas the other GCT members were not significantly detected in the heterocyst. The transcriptional responses of ANASP GCT genes to various oxidative stresses and growth environments were multifarious. Their intrinsic differences in transcriptional responsiveness and cellular localization suggest that this large GCT cluster is designed as an adaptive strategy to efficiently combat lipid hydroperoxide in Anabaena sp. that perform oxygenic photosynthesis and N(2) fixation. PMID- 17462543 TI - Hypoxia affects the physiological behavior of rat cortical synaptosomes. AB - Nerve cells, especially synaptosomes, are very susceptible to hypoxia and the subsequent oxidative stress. In this paper, we examined the effects of hypoxia (93% N(2):2% O(2):5% CO(2), v/v/v) on rat cortical synaptosomes by evaluating modifications of synaptosomal mitochondrial respiration rate and ATP production, membrane potential, intrasynaptosomal mitochondrial Ca(2+) concentration ([Ca(2+)](i)), and desferoxamine-chelatable free iron and esterified F2 isoprostane levels after different periods of hypoxia and after 30 min of reoxygenation. Oxygen consumption decreased significantly during 120 min of hypoxia and was restored after reoxygenation. At the same time, ATP production decreased and remained significantly lower even after reoxygenation. This involved a depolarization of the synaptosomal mitochondrial membrane, although the [Ca(2+)](i) remained practically unchanged. Indeed, iron and F2-isoprostane levels, representing useful prediction markers for neurodevelopmental outcome, increased significantly after hypoxia, and there was a strong correlation between the two variables. On the whole our results indicate that synaptosomal mitochondria undergo mitoptosis after 2 h of hypoxia. PMID- 17462544 TI - Physical activity and breast cancer risk among women in the southwestern United States. AB - Physical activity may influence breast cancer risk through multiple mechanisms and at different periods in life. In this study we evaluate breast cancer risk associated with total and vigorous physical activity at ages 15, 30, and 50 years and the referent year prior to diagnosis/selection. Participants were non Hispanic white (NHW) (1527 cases and 1601 control subjects) and Hispanic/American Indian (HAI) (798 cases and 924 controls) women. Both total and vigorous activity reduced risk of breast cancer in a dose-response manner. Among premenopausal women, only high total metabolic equivalent of the task (MET) hours of activity during the referent year was associated with reduced breast cancer risk in NHW women (odds ratio [OR] 0.62; 95% confidence interval [CI] 0.43, 0.91). Among postmenopausal women, physical activity had the greatest influence among women not recently exposed to hormones. Among these women, high total lifetime activity reduced risk of breast cancer for both NHW (OR 0.60; 95% CI 0.36, 1.02; p trend 0.01) and HAI women (OR 0.52; 95% CI 0.23, 1.16; p trend 0.07). Additionally, high total MET hours of activity at age 30 years (OR 0.56; 95% CI 0.37, 0.85) and at age 15 years (OR 0.57; 95% CI 0.38, 0.88) reduced breast cancer risk among postmenopausal NHW women not recently exposed to hormones. Among HAI women, more recent activity performed during the referent year and at age 50 appeared to have the greatest influence on breast cancer risk. Among postmenopausal NHW women. there was a significant interaction between physical activity and hormone replacement therapy (p value, 0.01), while among postmenopausal HAI women, physical activity interacted with body mass index (p value, 0.04). These data suggest that physical activity is important in reducing risk of breast cancer in both NHW and HAI women. PMID- 17462545 TI - The joint effect of oxidative stress and antioxidants on the risk of an aneurysmal rupture subarachnoid hemorrhage: a case-control study in Japan. AB - PURPOSE: A case-control study was conducted in Japan to examine the joint effects of oxidative stress and antioxidants on the risk of subarachnoid hemorrhage (SAH). METHODS: The study included 201 consecutive patients with first spontaneous SAH aged 30 to 79 years who had aneurysm(s) confirmed by angiography and/or computed tomographic scan and were admitted to two medical hospitals in Nagoya, Japan, from April 1992 to March 1997. Community control subjects were identified and matched to each case by gender and age (+/-2 years). Conditional logistic regression modeling was used to compute odds ratios (ORs) adjusted for body mass index, physical activity, family history of SAH, a history of hypertension, drinking habit, and frequency of green-yellow vegetables and fruits, and tea consumption. RESULTS: The greatest risk for SAH was posed by combination of current smoking and a less frequent intake of soy products (adjusted OR = 5.3; 95% confidence interval [CI], 2.0 to 14.3). Less frequent intake of soy products significantly elevated SAH risk, independent of smoking habits. CONCLUSION: These data suggested that enhancement of the antioxidant defense system such as much intake of soy products may be important in preventing SAH rather than the reduction of such oxidants as smoking. Our findings should prove useful for targeting individuals/populations in programs for primary prevention of SAH. PMID- 17462546 TI - Osteoarthritis: a comorbid marker for longer life? AB - PURPOSE: Diseases are often described and studied in isolation, yet there is increasing recognition of the complex interrelatedness of diseases and treatments in patients with multiple chronic diseases. Our objective was to describe the impact of selected diseases involving chronic inflammation (chronic obstructive pulmonary disease [COPD], osteoarthritis, and rheumatoid arthritis) on mortality. METHODS: We identified a cohort aged 55 to 64 years with one or more chronic conditions. Clusters of mutually exclusive disease combinations were created. Five-year all-cause mortality was determined and the relative risk (RR) of mortality was estimated when COPD, osteoarthritis, and rheumatoid arthritis were added to clusters. RESULTS: In 741,847 persons the 5-year mortality rates were lowest among persons with one condition and increased with more chronic conditions. The presence of osteoarthritis in a cluster was an exception where the risk was lower compared with that cluster without osteoarthritis: COPD (RR = 0.73 [95% confidence interval (CI), 0.65, 0.81]); ischemic heart disease (0.63 [0.52, 0.76]); hypertension (0.77 [0.71, 0.83]); dementia (0.63 [0.42, 0.93]); depression (0.65 [0.50, 0.84]); hypertension plus diabetes (0.85 [0.77, 0.93]); and ischemic heart disease plus hypertension (0.83 [0.73, 0.94]). CONCLUSIONS: The association between osteoarthritis and lower rates of mortality is notable and replicating these findings to explore causal relationships is important. PMID- 17462547 TI - Hypothesis: oxidative stress score as a combined measure of pro-oxidant and antioxidant exposures. AB - PURPOSE: To explore the hypothesis that a combination of several risk factors acting through the same pathway may produce an overall large increase in risk even in the presence of weak associations with each individual factor. METHODS: Using oxidative stress pathway as an example, we propose an oxidative stress score (OSS), where high and low pro-oxidant exposures expressed as continuous variables are assigned values of 0 and 1, while high and low antioxidant exposures are assigned values of 1 and 0, respectively. Dichotomous variables for pro-oxidant and antioxidant exposures are scored in a similar fashion. All individual scores are then summed to calculate the overall OSS, where higher and lower values indicate a shift toward antioxidant and pro-oxidant exposures, respectively. RESULTS: We illustrate this approach by using data from two previously-conducted case-control studies: a colonoscopy-based colorectal adenoma study, and a population-based prostate cancer study. In this pilot illustration we found a substantial decrease in risk associated with a high OSS for both prostate cancer and colorectal adenoma. By contrast, analyses for individual OSS components demonstrated no discernible pattern. CONCLUSIONS: Our exploratory analyses serve as a demonstration of a method and warrant further confirmation on a larger scale. PMID- 17462548 TI - Kidney growth during catabolic illness: what it does not destroy makes it grow stronger. AB - The kidney undergoes hypertrophy under conditions that paradoxically cause a loss of lean body mass, such as diabetes, acidosis, and chronic kidney disease. What unique mechanisms account for kidney growth during negative nitrogen balance? One adaptation is that renal tubular cells substantially decrease protein breakdown during kidney cell growth. In this review, we discuss how acidosis and diabetes reduce protein breakdown within the kidney and the intracellular signaling pathways that may regulate protein metabolism. Our results suggest that in cell culture models and in acute diabetes, kidney cells specifically reduce protein breakdown by the lysosomal pathway of chaperone-mediated autophagy. This differs from the activation of proteolysis by the ubiquitin-proteasome system in muscle in acute diabetes and uremia. A shared signaling pathway regulates protein breakdown in both kidney and skeletal muscle, namely, phosphatidylinositol-3 kinase signaling. Diabetes mellitus activates signaling through this pathway in the kidney while down-regulating it in skeletal muscle. We conclude that similar signaling pathways may regulate distinct proteolytic pathways in different tissues. PMID- 17462549 TI - Nutritional effect of nandrolone decanoate in predialysis patients with chronic kidney disease. AB - OBJECTIVE: The study objective was to examine the nutritional effect of nandrolone decanoate, an androgen derivative, in predialysis patients with chronic kidney disease (CKD). DESIGN: This was a prospective and experimental study. SETTING: The study was performed at the institutional level of clinical care. PATIENTS: Twenty-nine predialysis patients with CKD, with a glomerular filtration rate between 5 and 30 mL/min and moderate to severe malnutrition, were included and randomly divided into control (n = 13) and nandrolone decanoate (NAN, n = 16) groups. INTERVENTION: Patients in the control group received optimally conventional treatment of CKD. Patients in the NAN group, in addition to the conventional treatment, were intramuscularly injected with nandrolone decanoate at the dose of 100 mg per for 3 months. MAIN OUTCOME MEASURE: Nutritional markers, including lean body mass (LBM), normalized protein catabolic rate, serum albumin, and lipids, were determined at baseline and 3-month periods. RESULTS: Baseline parameters in both groups were not different. After 3 months, the patients in the NAN group had increased LBM (P < .01) and decreased serum albumin levels (P < .05), but no changes in the values of normalized protein catabolic rate, serum lipids, hematocrit, and glomerular filtration rate. No alterations in all parameters were identified in the control group. Changes in LBM in the NAN group were significantly higher than in the control group (P < .05). Minor adverse effects were observed in a few patients in the NAN group. CONCLUSION: Nandrolone decanoate expresses an anabolic effect on LBM without altering the renal function and thus would provide nutritional benefit in predialysis patients with CKD. PMID- 17462550 TI - Effects of a supplemented hypoproteic diet in chronic kidney disease. AB - OBJECTIVE: We assessed the effect of a severe hypoproteic diet supplemented with ketoanalogues (SVLPD) for 48 weeks on certain metabolic disorders of chronic kidney disease (CKD). DESIGN: We performed a prospective, open-label, parallel, randomized, controlled trial. SETTING: The study took place in the Nephrology Department at the Dr Carol Davila Teaching Hospital of Nephrology, Bucharest, Romania. PATIENTS: A total of 53 nondiabetic patients with CKD with an estimated glomerular filtration rate less than 30 mL/min/1.73 m(2) (Modification of Diet in Renal Disease formula), proteinuria less than 1 g/g urinary creatinine, good nutritional status, and anticipated good compliance with the diet were randomly assigned to two groups. INTERVENTION: Group I (n = 27) received the SVLPD (0.3 g/kg/d of vegetable proteins and ketoanalogues, 1 capsule for every 5 kg of ideal body weight per day). Group II (n = 26) continued a conventional low mixed protein diet (0.6 g/kg/d). OUTCOME MEASURES: Nitrogen waste products retention and calcium-phosphorus and acid-base disturbances were primary efficacy parameters, and "death" of the kidney or the patient and the estimated glomerular filtration rate were secondary efficacy parameters. The nutritional status and compliance with the diet were predefined as safety variables. There were no differences between groups in any parameter at baseline. RESULTS: In the SVLPD group, serum urea significantly decreased (56 +/- 7.9 mmol/L vs. 43.2 +/- 10 mmol/L), and significant improvements in serum bicarbonate (23.4 +/- 2.1 mmol/L vs. 18.1 +/- 1.5 mmol/L), serum calcium (1.10 +/- 0.17 mmol/L vs. 1.00 +/- 0.15 mmol/L at baseline), serum phosphates (1.45 +/- 0.66 mmol/L vs. 1.91 +/- 0.68 mmol/L), and calcium-phosphorus product (1.59 +/- 0.11 mmol(2)/L(2) vs. 1.91 +/- 0.10 mmol(2)/L(2)) were noted after 48 weeks. No death was registered in any group. Significantly lower percentages of patients in group I required renal replacement therapy initiation (4% vs. 27%). After 48 weeks, estimated glomerular filtration rate did not significantly change in patients receiving SVLPD (0.26 +/ 0.08 mL/s vs. 0.31 +/- 0.08 mL/s at baseline), but significantly decreased in controls (0.22 +/- 0.09 mL/s vs. 0.30 +/- 0.07 mL/s). The compliance with the keto-diet was good in enrolled patients. No significant changes in any of the parameters of the nutritional status and no adverse reactions were noted. CONCLUSION: SVLPD seems to ameliorate the nitrogen waste products retention and acid-base and calcium-phosphorus metabolism disturbances and to postpone the renal replacement therapy initiation, preserving the nutritional status in patients with CKD. PMID- 17462551 TI - Evaluation of nutrition assessment tools compared with body cell mass for the assessment of malnutrition in chronic kidney disease. AB - OBJECTIVE: We compared the subjective global assessment (SGA) and a range of SGA based assessment tools with body cell mass (BCM) in patients with stage IV and V predialysis chronic kidney disease (CKD). STUDY DESIGN: This was a cross sectional, observational study. SETTING: The study took place at a public tertiary hospital predialysis outpatient clinic. PATIENTS: A total of 56 consecutive consenting patients with CKD (61% were male; age [mean +/- standard deviation] 70.2 +/- 11.6 years; glomerular filtration rate 22.2 +/- 6.8 mL/min). MAIN OUTCOME MEASURE: Nutrition status was the main outcome measure. RESULTS: In this population, the prevalence of malnutrition was 19.6% (n = 11, SGA B; no C ratings). Malnutrition was associated with lower BCM (mean BCM, 26.3 vs. 33.4 kg, P = .007, measured by total body potassium), body weight (64.8 vs. 76.1 kg, P = .042), body mass index (23.7 vs. 27.6 kg/m(2), P = .015), and greater weight loss over the previous 6 months (-6.2 vs. -0.1 kg, P = .004). BCM had a weak relationship with 7-point SGA (P = .267), malnutrition inflammation score (r = 0.27 P = .063), and patient-generated SGA (r = -0.27 P = .060). There was no association for either measure of nutrition status (SGA or BCM) with albumin, glomerular filtration rate, or C-reactive protein. CONCLUSION: SGA in its original form most accurately delineated malnutrition by depleted BCM and is the most appropriate tool for cross-sectional assessment of nutrition status in patients with predialysis CKD. PMID- 17462552 TI - Body composition and physical activity in end-stage renal disease. AB - OBJECTIVE: The study objective was to examine the relationship between visceral and somatic protein stores and physical activity in individuals with end-stage renal disease. DESIGN: This was a prospective single-center study. SETTING: The study took place at the Vanderbilt University Outpatient Dialysis Unit and General Clinical Research Center. PATIENTS: Fifty-five patients with prevalent chronic hemodialysis (CHD) were included: 33 males, 22 females, 45 African Americans, 9 Caucasians, and 1 Asian. The mean age was 47.0 +/- 1.6 years, height was 166.4 +/- 13.9 cm, and weight was 83.1 +/- 2.6 kg. METHODS: Body composition was measured by dual-energy x-ray absorptiometry. Minute-by-minute physical activity was assessed over a 7-day period with a triaxial accelerometer. Participants were interviewed by a trained registered dietitian for two 24-hour diet recalls (one from a hemodialysis day; one from a nonhemodialysis day). Laboratory values for serum concentrations of albumin, prealbumin, C-reactive protein, and creatinine were also collected. MAIN OUTCOME MEASURE: Predictors of somatic protein stores were the main outcome measure. RESULTS: Serum albumin was negatively and significantly correlated with the percentage of fat mass (P = .016) and kg of fat mass (P = .044). C-reactive protein was positively and significantly correlated with body weight (P = .006), percentage of fat mass (P = .017), kg of fat mass (P = .006), and body mass index (P = .004). Physical activity and total daily protein intake were the strongest predictors of the amount of lean body mass (P = .01 and .003, respectively). CONCLUSION: The association between somatic protein and visceral protein stores is weak in patients with CHD. Whereas increased levels of physical activity and total daily protein intake are associated with higher lean body mass in patients with CHD, higher adiposity is associated with higher C-reactive protein and lower albumin values. PMID- 17462553 TI - Effect of a sardine supplement on C-reactive protein in patients receiving hemodialysis. AB - OBJECTIVE: The study evaluated the effect of a canned sardine supplement in C reactive protein (CRP) in patients on hemodialysis (HD) and the compliance and adherence to this supplement. DESIGN: This was a quasi-experimental study: Participants with a serum CRP of 5 mg/dL or less volunteered to consume a sardine supplement or were maintained on the usual cheese/ham sandwich supplement. SETTING: The study took place in two outpatient dialysis units in Lisbon, Portugal. PATIENTS: The study comprised 63 patients receiving maintenance HD three times per week for at least 6 months and an initial CRP concentration of 5 mg/dL or less. Exclusion criteria included the presence of graft vascular access or history of cancer. INTERVENTION: After a 4-week washout period, the nutritional intervention included a canned sardine sandwich for the case group (n = 31) and a cheese or ham sandwich for the control group (n = 32), to be ingested during each routine HD session, 3 times per week, for 8 weeks. MAIN OUTCOME MEASURE: Serum levels of high-sensitivity CRP were the outcome measure. RESULTS: Only 65 patients from the invited 186 patients met the inclusion criteria and agreed to eat the sardine sandwich supplement three times per week and were involved in the study. A significant proportion of 48% (n = 31, case group) consumed the sardine sandwich supplement three times per week for 8 weeks, fulfilling the requirements and completing the study. The present investigation showed that a sardine sandwich supplement had no effect on CRP levels among patients on HD. However, when participants were stratified according to tertiles of CRP distribution values at baseline, a reduction in CRP levels was found for those in the higher tertile, being higher for the case group (P = .047). Although diabetic patients were excluded from the analysis (eight in the sardine supplementation group and seven in the control group) a significant CRP reduction was found (P = .034). CONCLUSION: Although a supplement of low-dose n-3 long chain polyunsaturated fatty acids had no effect on the plasma high-sensitivity CRP of the supplemented group, a reduction in CRP levels was found when patients were stratified for tertiles of CRP (for the upper tertile) and diabetic status (for nondiabetic patients). These findings need to be further confirmed. This canned sardine supplement was accepted by an important proportion of patients, enhancing diet variety and contributing for a greater n-3 long-chain polyunsaturated fatty acids eicosapentaenoic acid and docosahexaenoic acid intake. PMID- 17462554 TI - Protein supplementation with sports protein bars in renal patients. AB - BACKGROUND: Malnutrition prevalence in patients on dialysis is well established. The protein requirements for both hemodialysis and peritoneal dialysis have been documented elsewhere, including the Kidney Disease Outcomes Quality Initiative Clinical Practice Guidelines for Nutrition in Chronic Renal Failure. The clinical challenge is to assist patients in meeting these targets, especially in those with anorexia. Traditional supplements have included fluid, which is an issue for patients who are fluid restricted. OBJECTIVES: The study objectives were to (1) investigate the range of sports protein supplements that may be suitable for patients on hemodialysis to use and (2) trial nonfluid protein supplements in patients on hemodialysis. METHODS AND DESIGN: Known manufacturers of sports protein bars and other sports supplements available in Australia were contacted for the nutrient breakdown of high-protein products, specifically potassium, protein, and phosphorus contents. As a result, selected high-protein sports bars (Protein FX, Aussie Bodies, Port Melbourne, Victoria, Australia) were used as an alternative to the more commonly used renal-specific fluid supplements (Nepro, Abbott Laboratories, Abbott Park, IL; Novasource Renal, Novartis Nutrition Corporation, Fremont, MI; and Renilon, Nutricia, Wiltshire, UK) in patients with poor nutritional status requiring supplementation. Patient satisfaction and clinical nutrition markers were investigated. SETTING: The study took place at inpatient, in-center, and satellite hemodialysis settings in Adelaide, South Australia. PARTICIPANTS: A total of 32 patients (16 females and 16 males) with an average age of 62.9 years (range 32-86 years) undergoing hemodialysis (acute and maintenance) were included. Subjects were selected by the author as part of routine clinical nutrition care. INTERVENTION: Patients trialed sports protein bars as a protein supplement alone or in conjunction with other supplementary products. RESULTS: All patients were in favor of the trial, with 22 of 32 patients continuing with the protein bars as the preferred supplement. The major reasons for not continuing were taste and chewability, especially in older patients with dentures. Serum potassium and phosphate levels were not increased with supplementation. Measured serum albumin and protein catabolic rate were considered unreliable indicators because not all patients were medically stable. CONCLUSIONS: Sports protein bars are an acceptable protein and energy supplement for patients on hemodialysis. Sports protein bars are well accepted by patients except when dentures limit chewability. Sports protein bars have advantages over fluid-based supplements in patients with fluid restrictions. PMID- 17462555 TI - Notes from the CKD kitchen: a variety of salt-free seasonings. AB - One of the challenges with renal diets is how to make flavorful food while maintaining the low sodium restrictions. I have found three spice companies that have created seasoning mixtures using a mixture of herbs that do not add sodium or potassium chloride in their flavors. The recipe Ginger Roasted Chicken with an Asian Slaw is an example of how you can use three different salt-free seasonings to create a flavorful meal. I know from personal experience that I feel better, have more energy, and sleep better if I restrict my sodium intake. It is easy to stop using the salt shaker and replace the garlic salt and onion salt with garlic powder and onion powder in the kitchen. It takes a dedicated shopper to find the hidden sodium in foods. I find myself reading more and more labels in the aisles of the grocery store before I put any foods in my grocery cart. I also find myself studying the spice selections looking for salt-free seasonings. Mrs. Dash is great and very popular, but there must be more options for us patients with chronic kidney disease. After doing some culinary research, I was pleased to find a much larger section of salt-free seasonings than I expected. I have listed a few of the seasoning combinations below and a table of three major spice companies along with their contact information for purchasing their products. PMID- 17462556 TI - Dialysis laughing. PMID- 17462559 TI - Pulmonary vascular remodeling. AB - The maladaptive response of the pulmonary vasculature that occurs in patients with congenital diaphragmatic hernia significantly impacts outcome. Muscularized distal pulmonary arterioles inhibit the ability of the neonate to adjust to extrauterine circulation, resulting in severe pulmonary hypertension. This review summarizes the current state of knowledge regarding normal and abnormal development of the lung vascular system and identifies current and potential therapies directed toward preserving or restoring proper pulmonary vascular function. PMID- 17462560 TI - Genetics of congenital diaphragmatic hernia. AB - Congenital diaphragmatic hernia (CDH) is a common structural birth defect that affects approximately 1 in 2500 live births. Although the exact etiology of most cases of CDH remains unknown, it is becoming increasingly clear that genetic factors play an important role in many cases of CDH. In this paper, we review critical findings in the areas of clinical and basic research that highlight the importance of genetics in the development of CDH. We also provide practical information that can aid physicians and surgeons as they evaluate and care for patients with isolated, nonisolated, and syndromic forms of CDH and their families. PMID- 17462561 TI - Diaphragm development and congenital diaphragmatic hernia. AB - Advances in the understanding of normal diaphragm embryogenesis have provided the necessary foundation for novel insights into the pathogenesis of congenital diaphragmatic hernia (CDH). Although diaphragm formation is still not completely understood, we have identified key structures and periods of development that are clearly abnormal in animal models of CDH. The pleuroperitoneal fold (PPF) is a transient structure which is the target for the neuromuscular component of the diaphragm. The PPF has been shown to be abnormal in multiple animal models of Bochdalek CDH; specifically, a malformation of the nonmuscular component of this tissue is thought to underlie the later defect in the complete diaphragm. Based on data from animal models and the examination of human postmortem tissue, we hypothesize that abnormal PPF development underlies Bochdalek CDH. Further, the understanding of the pathogenesis of rarer subtypes of CDH will be advanced by the study of various new animal models discussed in this review. PMID- 17462562 TI - Prenatal intervention for congenital diaphragmatic hernia. AB - Advances in prenatal ultrasound have revealed the poor natural history of fetal congenital diaphragmatic hernia (CDH) and its hidden mortality during gestation and immediately after birth. Attempts to improve this poor outcome led to the development of prenatal surgical intervention for severe CDH by Harrison and his colleagues at the University of California San Francisco. Prenatal surgical intervention for CDH has seen four phases: open fetal surgical repair, open surgical tracheal occlusion, endoscopic external tracheal occlusion, and endoscopic endoluminal tracheal occlusion. After extensive work in the laboratory, prenatal intervention has been applied in humans since 1984. With the most recent techniques, maternal risk is significantly reduced as is the incidence of preterm labor. In the meantime, the survival rate of fetuses with CDH without fetal intervention has improved mainly due to the minimization of iatrogenic lung injury by gentle ventilation, first described in 1985. However, the morbidity of the survivors with severe CDH remains high. Prenatal intervention for CDH will be justified if improvement in survival or morbidity can be demonstrated when compared to planned delivery and postnatal management with gentle ventilation strategy. PMID- 17462563 TI - Surgical management of neonates with congenital diaphragmatic hernia. AB - Congenital diaphragmatic hernia (CDH) is one of the most challenging and complex pediatric abnormalities to manage, both medically and surgically. The care of these neonates has seen significant evolution, from previous aggressive ventilation and emergent operation to current permissive hypercapnea, physiologic stabilization, and elective surgical repair, all in less than two decades. These changes have led to many centers reporting survival rates near 80%, a dramatic improvement from the 50% survival reported in the 1970s. This review covers the current principles guiding the surgical management of CDH in the neonate, including preoperative stabilization, operative timing, extracorporeal membrane oxygenation, surgical approach, and management of recurrence. Although many clinical challenges remain, multi-institutional collaboration and ongoing research efforts will hopefully improve the clinical care of these patients. PMID- 17462564 TI - Mechanical ventilation strategies in the management of congenital diaphragmatic hernia. AB - Most infants with congenital diaphragmatic hernia (CDH) require respiratory support. The goal of this report is to present an overview of mechanical ventilation strategies in the management of infants with CDH. The anatomic and physiologic limitations in the lungs of infants with diaphragmatic hernia make decisions on the best strategy and use of mechanical ventilation challenging. We will briefly review lung development in infants with CDH, identifying factors that provide a basis for lung protection strategies. Background on the use of specific mechanical ventilation modes and the rationale for each are provided. Finally, we review mechanical ventilation practices described in published case series of successful CDH management, with a brief review of additional treatments, including inhaled nitric oxide and extracorporeal membrane oxygenation. Although details of a single specific best strategy for mechanical ventilation for CDH infants cannot be identified from current literature, a lung protection ventilation approach, regardless of the device used, appears to reduce mortality risk. PMID- 17462565 TI - Pulmonary hypertension in congenital diaphragmatic hernia. AB - Clinically significant pulmonary hypertension (PHTN) is a common finding in newborn infants with congenital diaphragmatic hernia (CDH) resulting in right to left shunting at pre- and postductal level, hypoxemia, and acute right heart failure in those most severely affected. Even in those without clinical manifestations of ductal shunting, cardiac echo studies would suggest that increased pulmonary vascular resistance and right ventricular pressures are almost a universal finding in this disease, and in some instances, may persist well into the postnatal period. The lung is small and structurally abnormal, and the pulmonary vascular bed is not only reduced in size, but responds abnormally to vasodilators. During the last 20 years, "gentle" ventilation, delayed surgery, and improved peri-operative care have made the greatest impact in decreasing mortality in this condition. Use of PGE1 should be considered early if there is hemodynamically significant PHTN, right ventricular dysfunction, and the patent ductus arteriosus (PDA) is becoming restrictive. In individual patients, inhaled nitric oxide (iNO) might be helpful, but the response to iNO should be confirmed using echocardiography. In patients who survive operation and leave the hospital, there are chronic causes of morbidity that need to be looked for and managed in a multi-disciplinary follow-up clinic. PMID- 17462566 TI - Long-term follow up of infants with congenital diaphragmatic hernia. AB - With improving treatment strategies for congenital diaphragmatic hernia (CDH) infants, an increase in survival of more severely affected patients can be expected. Consequently, more attention is now focused on long-term follow up of these patients. Many reports have emphasized associated morbidity, including pulmonary sequelae, neurodevelopmental deficits, gastrointestinal disorders, and other abnormalities. Therefore, survivors of CDH remain a complex patient population to care for throughout infancy and childhood, thus requiring long-term follow up. Much information has been provided from many centers regarding individual institutional improvements in overall survival. Few of these, however, have reported long-term follow up. The aim of this review is to describe the long term outcome of survivors with CDH and to suggest a possible follow-up protocol for these patients. PMID- 17462567 TI - Neuropathic pain treatment gives Avigen new lease on life and Neuromed follows the snail trail. PMID- 17462568 TI - A direct inhibitor of HMGB1 cytokine. AB - HMGB1 and the recently described interleukin-33 (IL-33) are abundant chromatin associated nuclear factors with potent proinflammatory cytokine activities. In this issue of Chemistry & Biology, Mollica et al. [1] report the identification of the first direct small-molecule inhibitor of HMGB1. PMID- 17462569 TI - Chemical tools from biology-oriented synthesis. AB - Lessmann et al. present a chiral natural-product-derived library of alpha,beta unsaturated delta-lactones obtained through stereoselective synthesis on a solid support [1]. Phenotypic screening of this compound collection provided new modulators of cell cycle progression and of viral cell entry with high hit rates. PMID- 17462570 TI - Rapid identification of antibacterial agents effective against Staphylococcus aureus using small-molecule macroarrays. AB - There is an urgent, global need for the development of new antibacterial agents. We have applied the small-molecule macroarray approach to the synthesis and screening of antibacterial compounds active against the Gram-positive pathogen Staphylococcus aureus. Several macroarrays of 1,3-diphenyl-2-propen-1-ones (chalcones), cyanopyridines, and pyrimidines were synthesized on a planar cellulose support system on the order of days. This support system was found to be highly compatible with antibacterial assay formats, including disk-diffusion and agar-overlay visualization methods. Further, sufficient compound was isolated from each spot of the macroarray for both compound characterization and minimum inhibitory concentration (MIC) estimation. Analysis of the small-molecule macroarrays in these assays uncovered a set of antibacterial agents with in vitro MIC values against methicillin-resistant S. aureus comparable to certain antibacterial drugs in use today. PMID- 17462571 TI - Structural insight into chain-length control and product specificity of pentaketide chromone synthase from Aloe arborescens. AB - The crystal structures of a wild-type and a mutant PCS, a novel plant type III polyketide synthase from a medicinal plant, Aloe arborescens, were solved at 1.6 A resolution. The crystal structures revealed that the pentaketide-producing wild type and the octaketide-producing M207G mutant shared almost the same overall folding, and that the large-to-small substitution dramatically increases the volume of the polyketide-elongation tunnel by opening a gate to two hidden pockets behind the active site of the enzyme. The chemically inert active site residue 207 thus controls the number of condensations of malonyl-CoA, solely depending on the steric bulk of the side chain. These findings not only provided insight into the polyketide formation reaction, but they also suggested strategies for the engineered biosynthesis of polyketides. PMID- 17462572 TI - Identification of a small-molecule inhibitor of class Ia PI3Ks with cell-based screening. AB - The mammalian target of rapamycin (mTOR) signaling network is central to the regulation of cell growth in response to both growth factors and nutrients. We developed a high-throughput, cell-based assay to identify small-molecule modulators of the mTOR signaling network. One such compound, which we name quinostatin, potently inhibits this network by directly targeting the lipid kinase activity of the catalytic subunits of class Ia PI3Ks. This study illustrates the power of unbiased, phenotypic screening as a means for illuminating cell circuitry, and resulted in the identification of a chemotype for selective inhibition of the class Ia PI3Ks. PMID- 17462573 TI - Biosynthesis of butirosin: transfer and deprotection of the unique amino acid side chain. AB - Butirosin, an aminoglycoside antibiotic produced by Bacillus circulans, bears the unique (S)-4-amino-2-hydroxybutyrate (AHBA) side chain, which protects the antibiotic from several common resistance mechanisms. The AHBA side chain is advantageously incorporated into clinically valuable antibiotics such as amikacin and arbekacin by synthetic methods. Therefore, it is of significant interest to explore the biosynthetic origins of this useful moiety. We report here that the AHBA side chain of butirosin is transferred from the acyl carrier protein (ACP) BtrI to the parent aminoglycoside ribostamycin as a gamma-glutamylated dipeptide by the ACP:aminoglycoside acyltransferase BtrH. The protective gamma-glutamyl group is then cleaved by BtrG via an uncommon gamma-glutamyl cyclotransferase mechanism. The application of this pathway to the in vitro enzymatic production of novel AHBA-bearing aminoglycosides is explored with encouraging implications for the preparation of unnatural antibiotics via directed biosynthesis. PMID- 17462574 TI - High-density targeting of a viral multifunctional nanoplatform to a pathogenic, biofilm-forming bacterium. AB - Nanomedicine directed at diagnosis and treatment of infections can benefit from innovations that have substantially increased the variety of available multifunctional nanoplatforms. Here, we targeted a spherical, icosahedral viral nanoplatform to a pathogenic, biofilm-forming bacterium, Staphylococcus aureus. Density of binding mediated through specific protein-ligand interactions exceeded the density expected for a planar, hexagonally close-packed array. A multifunctionalized viral protein cage was used to load imaging agents (fluorophore and MRI contrast agent) onto cells. The fluorescence-imaging capability allowed for direct observation of penetration of the nanoplatform into an S. aureus biofilm. These results demonstrate that multifunctional nanoplatforms based on protein cage architectures have significant potential as tools for both diagnosis and targeted treatment of recalcitrant bacterial infections. PMID- 17462575 TI - Conotoxins containing nonnatural backbone spacers: cladistic-based design, chemical synthesis, and improved analgesic activity. AB - Disulfide-rich neurotoxins from venomous animals continue to provide compounds with therapeutic potential. Minimizing neurotoxins often results in removal of disulfide bridges or critical amino acids. To address this drug-design challenge, we explored the concept of disulfide-rich scaffolds consisting of isostere polymers and peptidic pharmacophores. Flexible spacers, such as amino-3 oxapentanoic or 6-aminohexanoic acids, were used to replace conformationally constrained parts of a three-disulfide-bridged conotoxin, SIIIA. The peptide polymer hybrids, polytides, were designed based on cladistic identification of nonconserved loci in related peptides. After oxidative folding, the polytides appeared to be better inhibitors of sodium currents in dorsal root ganglia and sciatic nerves in mice. Moreover, the polytides appeared to be significantly more potent and longer-lasting analgesics in the inflammatory pain model in mice, when compared to SIIIA. The resulting polytides provide a promising strategy for transforming disulfide-rich peptides into therapeutics. PMID- 17462576 TI - Spin-labeled analogs of CMP-NeuAc as NMR probes of the alpha-2,6 sialyltransferase ST6Gal I. AB - Structural data on mammalian proteins are often difficult to obtain by conventional NMR approaches because of an inability to produce samples with uniform isotope labeling in bacterial expression hosts. Proteins with sparse isotope labels can be produced in eukaryotic hosts by using isotope-labeled forms of specific amino acids, but structural analysis then requires information from experiments other than nuclear Overhauser effects. One source of alternate structural information is distance-dependent perturbation of spin relaxation times by nitroxide spin-labeled analogs of natural protein ligands. Here, we introduce spin-labeled analogs of sugar nucleotide donors for sialyltransferases, specifically, CMP-TEMPO (CMP-4-O-[2,2,6,6-tetramethylpiperidine-1-oxyl]) and CMP 4carboxyTEMPO (CMP-4-O-[4-carboxy-2,2,6,6-tetramethylpiperidinine-1-oxyl]). An ability to identify resonances from active site residues and produce distance constraints is illustrated on a (15)N phenylalanine-labeled version of the structurally uncharacterized, alpha-2,6-linked sialyltransferase, ST6Gal I. PMID- 17462577 TI - LMP2-specific inhibitors: chemical genetic tools for proteasome biology. AB - The immunoproteasome, having been linked to neurodegenerative diseases and hematological cancers, has been shown to play an important role in MHC class I antigen presentation. However, its other pathophysiological functions are still not very well understood. This can be attributed mainly to a lack of appropriate molecular probes that can selectively modulate the immunoproteasome catalytic subunits. Herein, we report the development of molecular probes that selectively inhibit the major catalytic subunit, LMP2, of the immunoproteasome. We show that these compounds irreversibly modify the LMP2 subunit with high specificity. Importantly, LMP2-rich cancer cells compared to LMP2-deficient cancer cells are more sensitive to growth inhibition by the LMP2-specific inhibitor, implicating an important role of LMP2 in regulating cell growth of malignant tumors that highly express LMP2. PMID- 17462578 TI - Glycyrrhizin binds to high-mobility group box 1 protein and inhibits its cytokine activities. AB - High-mobility group box 1 protein (HMGB1) is a nuclear component, but extracellularly it serves as a signaling molecule involved in acute and chronic inflammation, for example in sepsis and arthritis. The identification of HMGB1 inhibitors is therefore of significant experimental and clinical interest. We show that glycyrrhizin, a natural anti-inflammatory and antiviral triterpene in clinical use, inhibits HMGB1 chemoattractant and mitogenic activities, and has a weak inhibitory effect on its intranuclear DNA-binding function. NMR and fluorescence studies indicate that glycyrrhizin binds directly to HMGB1 (K(d) approximately 150 microM), interacting with two shallow concave surfaces formed by the two arms of both HMG boxes. Our results explain in part the anti inflammatory properties of glycyrrhizin, and might direct the design of new derivatives with improved HMGB1-binding properties. PMID- 17462579 TI - Natural product-derived modulators of cell cycle progression and viral entry by enantioselective oxa Diels-Alder reactions on the solid phase. AB - The underlying frameworks of natural product classes with multiple biological activities can be regarded as biologically selected and prevalidated starting points in vast chemical structure space in the development of compound collections for chemical biology and medicinal chemistry research. For the synthesis of natural product-derived and -inspired compound collections, the development of enantioselective transformations in a format amenable to library synthesis, e.g., on the solid support, is a major and largely unexplored goal. We report on the enantioselective solid-phase synthesis of a natural product inspired alpha,beta-unsaturated delta-lactone collection and its investigation in cell-based screens monitoring cell cycle progression and viral entry into cells. The screens identified modulators of both biological processes at a high hit rate. The screen for inhibition of viral entry opens up avenues of research for the identification of compounds with antiviral activity. PMID- 17462580 TI - Examination of the role of intestinal fatty acid-binding protein in drug absorption using a parallel artificial membrane permeability assay. AB - Transcellular diffusion across the absorptive epithelial cells (enterocytes) of the small intestine is the main route of absorption for most orally administered drugs. The process by which lipophilic compounds transverse the aqueous environment of the cytoplasm, however, remains poorly defined. In the present study, we have identified a structurally diverse group of lipophilic drugs that display low micromolar binding affinities for a cytosolic lipid-binding protein - intestinal fatty acid-binding protein (I-FABP). Binding to I-FABP significantly enhanced the transport of lipophilic drug molecules across a model membrane, and the degree of transport enhancement was related to both drug lipophilicity and I FABP binding affinity. These data suggest that intracellular lipid-binding proteins such as I-FABP may enhance the membrane transport of lipophilic xenobiotics and facilitate drug access to the enterocyte cytoplasm and cytoplasmic organelles. PMID- 17462581 TI - Removal of polyethylene glycols from protein samples using titanium dioxide. PMID- 17462582 TI - Understanding the mechanisms of zinc-sensing by metal-response element binding transcription factor-1 (MTF-1). AB - The regulation of divalent zinc has been observed in a wide range of organisms. Since this metal is an essential nutrient, but also toxic in excess, zinc homeostasis is crucial for normal cellular functioning. The metal-responsive element-binding transcription factor-1 (MTF-1) is a key regulator of zinc in higher eukaryotes ranging from insects to mammals. MTF-1 controls the expression of metallothioneins (MTs) and a number of other genes directly involved in the intracellular sequestration and transport of zinc. Although the diverse functions of MTF-1 extend well beyond zinc homeostasis to include stress-responses to heavy metal toxicity, oxidative stress, and selected chemical agents, in this review we focus on the recent advances in understanding the mechanisms whereby MTF-1 regulates MT gene expression to protect the cell from fluctuations in environmental zinc. Particular emphasis is devoted to recent studies involving the Cys2His2 zinc finger DNA-binding domain of MTF-1, which is an important contributor to the zinc-sensing and metal-dependent transcriptional activation functions of this protein. PMID- 17462583 TI - Study of the interaction of an alpha-helical transmembrane peptide with phosphatidylcholine bilayer membranes by means of densimetry and ultrasound velocimetry. AB - We applied precise densimetry and ultrasound velocimetry methods to study the interaction of a synthetic alpha-helical transmembrane peptide, acetyl-K(2)-L(24) K(2)-amide (L(24)), with model bilayer lipid membranes. The large unilamellar vesicles (LUVs) utilized were composed of a homologous series of n-saturated diacylphosphatidylcholines (PCs). PCs whose hydrocarbon chains contained from 13 to 16 carbon atoms, thus producing phospholipid bilayers of different thicknesses and gel to liquid-crystalline phase transition temperatures. This allowed us to analyze how the difference between the hydrophobic length of the peptide and the hydrophobic thickness of the lipid bilayer influences the thermodynamical and mechanical properties of the membranes. We showed that the incorporation of L(24) decreases the temperature and cooperativity of the main phase transition of all LUVs studied. The presence of L(24) in the bilayer also caused an increase of the specific volume and of the volume compressibility in the gel state bilayers. In the liquid crystalline state, the peptide decreases the specific volume at relatively higher peptide concentration (mole ratio L(24):PC=1:50). The overall volume compressibility of the peptide-containing lipid bilayers in the liquid crystalline state was in general higher in comparison with pure membranes. There was, however, a tendency for the volume compressibility of these lipid bilayers to decrease with higher peptide content in comparison with bilayers of lower peptide concentration. For one lipid composition, we also compared the thermodynamical and mechanical properties of LUVs and large multilamellar vesicles (MLVs) with and without L(24). As expected, a higher cooperativity of the changes of the thermodynamical and mechanical parameters took place for MLVs in comparison with LUVs. These results are in agreement with previously reported DSC and (2)H NMR spectroscopy study of the interaction of the L(24) and structurally related peptides with phosphatidylcholine bilayers. An apparent discrepancy between (2)H NMR spectroscopy and compressibility data in the liquid crystalline state may be connected with the complex and anisotropic nature of macroscopic mechanical properties of the membranes. The observed changes in membrane mechanical properties induced by the presence of L(24) suggest that around each peptide a distorted region exists that involves at least 2 layers of lipid molecules. PMID- 17462584 TI - Substitution of the leucine zipper sequence in melittin with peptoid residues affects self-association, cell selectivity, and mode of action. AB - Melittin (ME), a non-cell-selective antimicrobial peptide, contains the leucine zipper motif, wherein every seventh amino acid is leucine or isolucine. Here, we attempted to generate novel cell-selective peptides by substituting amino acids in the leucine zipper sequence of ME with peptoid residues. We generated a series of ME analogues by replacing Leu-6, Lue-13 and Ile-20 with Nala, Nleu, Nphe, or Nlys, and we examined their secondary structure, self-association activity, cell selectivity and mode of action. Circular dichroism spectroscopy indicated that the substitutions disrupt the alpha-helical structure of ME in micelles of sodium dodecyl sulfate and on negatively charged and zwitterionic phospholipid vesicles. Substitution by Nleu, Nphe, or Nlys but not Nala disturbed the self-association in an aqueous environment, interaction with zwitterionic membranes, and toxicity to mammalian cells of ME but did not affect the interaction with negatively charged membranes or antibacterial activity. Notably, peptides with Nphe or Nlys substitution had the highest therapeutic indices, consistent with their lipid selectivity. In addition, all of peptoid residue-containing ME analogues had little or no ability to induce membrane disruption, membrane depolarization and lipid flip-flop. Taken together, our studies indicate that substitution of the leucine zipper motif in ME with peptoid residues increases its selectivity against bacterial cells by impairing self-association activity and changes its mode of antibacterial action from membrane-targeting mechanism to possible intracellular targeting mechanism. Furthermore, our ME analogues especially those with Nleu, Nphe, or Nlys substitutions, may be therapeutically useful antimicrobial peptides. PMID- 17462585 TI - Chemical specificity in short-chain fatty acids and their analogues in increasing osmotic fragility in rat erythrocytes in vitro. AB - We examined the role of the chemical specificity of short-chain fatty acids (SCFAs) and their derivatives in increasing osmotic fragility (OF) in rat red blood cells (RBCs). Except for formic acid, normal SCFAs with 2 to 8 carbons increased the OF in rat RBCs with increasing number of hydrocarbons in a dose dependent manner. Replacement of the carboxylic group with sulfonic group inhibited, but did not abolish, the SCFA-mediated increase in OF. Introduction of another carboxylic group (dicarboxylic acids) completely abolished the SCFA mediated increase in OF. Transformation of the hydrocarbon chains in SCFAs from straight to branched or cyclic chains affected the degree of the OF-increasing effect. Introduction of double- or triple-carbon bonds to the hydrocarbon chain in parent SCFAs did not affect the increase in OF. Both hydrophilic (carboxylic group) and hydrophobic elements (hydrocarbons) at opposite sides of a molecule were required to affect the RBC membrane, and the size and form of hydrophobic element were important factors in determining the SCFA-mediated increase in OF. The hydrocarbon chains probably enter the plasma membrane, with the hydrophilic carboxylic base remaining outside of the membrane, and interact with phospholipid in cell membrane and disturb the structure of lipid layer resulting in the increase in OF in the rat RBCs. PMID- 17462586 TI - Inhibition of malaria parasite blood stages by tyrocidines, membrane-active cyclic peptide antibiotics from Bacillus brevis. AB - Tyrothricin, a complex mixture of antibiotic peptides from Bacillus brevis, was reported in 1944 to have antimalarial activity rivalling that of quinine in chickens infected with Plasmodium gallinaceum. We have isolated the major components of tyrothricin, cyclic decapeptides collectively known as the tyrocidines, and tested them against the human malaria parasite Plasmodium falciparum using standard in vitro assays. Although the tyrocidines differ from each other by conservative amino acid substitutions in only three positions, their observed 50% parasite inhibitory concentrations (IC(50)) spanned three orders of magnitude (0.58 to 360 nM). Activity correlated strictly with increased apparent hydrophobicity and reduced total side-chain surface area and the presence of ornithine and phenylalanine in key positions. In contrast, mammalian cell toxicity and haemolytic activities of the respective peptides were considerably less variable (2.6 to 28 microM). Gramicidin S, a structurally analogous antimicrobial peptide, was less active (IC(50)=1.3 microM) and selective than the tyrocidines. It exerted its parasite inhibition by rapid and selective lysis of infected erythrocytes as judged by fluorescence and light microscopy. The tyrocidines, however, did not cause an overt lysis of infected erythrocytes, but an inhibition of parasite development and life-cycle progression. PMID- 17462587 TI - Fructans from oat and rye: composition and effects on membrane stability during drying. AB - Fructans have been implicated in the abiotic stress tolerance of many plant species, including grasses and cereals. To elucidate the possibility that cereal fructans may stabilize cellular membranes during dehydration, we used liposomes as a model system and isolated fructans from oat (Avena sativa) and rye (Secale cereale). Fructans were fractionated by preparative size exclusion chromatography into five defined size classes (degree of polymerization (DP) 3 to 7) and two size classes containing high DP fructans (DP>7 short and long). They were characterized by high performance liquid chromatography (HPLC) and matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS). The effects of the fructans on liposome stability during drying and rehydration were assessed as the ability of the sugars to prevent leakage of a soluble marker from liposomes and liposome fusion. Both species contain highly complex mixtures of fructans, with a DP up to 17. The two DP>7 fractions from both species were unable to protect liposomes, while the fractions containing smaller fructans were protective to different degrees. Protection showed an optimum at DP 4 and the DP 3, 4, and 5 fractions from oat were more protective than all other fractions from both species. In addition, we found evidence for synergistic effects in membrane stabilization in mixtures of low DP with DP>7 fructans. The data indicate that cereal fructans have the ability to stabilize membranes under stress conditions and that there are size and species dependent differences between the fructans. In addition, mixtures of fructans, as they occur in living cells may have protective properties that differ significantly from those of the purified fractions. PMID- 17462588 TI - Effectiveness of treatment of childhood memories in cognitive therapy for personality disorders: a controlled study contrasting methods focusing on the present and methods focusing on childhood memories. AB - This study tested the hypothesis that treatment of childhood memories is an effective way to change personality disorder related schemas and psychopathology in cognitive therapy for personality disorders. To test this hypothesis, a crossover design was used to compare the effectiveness of methods focusing on the present and methods focusing on childhood memories. After the exploration period, the therapist focused either first on the present during 24 sessions and than for 24 sessions on childhood memories, or followed the reverse order. Twenty-one patients with one or more Axis II disorders were included. Participants were randomly assigned to order of focus. Results indicate that CT methods focusing on childhood memories produce good outcomes, comparable to those of methods focusing on the present. There was no significant effect of order, but both patients and therapists preferred the past-present order. Total effects of the package were large and were maintained till one-year follow-up (d's 0.97-1.90). Experience of the therapist with CT for personality disorders was related to better outcome (d=0.73). PMID- 17462589 TI - Enhanced perceived responsibility decreases metamemory but not memory accuracy in obsessive-compulsive disorder (OCD). AB - Mixed findings have been obtained in prior research with respect to the presence and severity of memory and metamemory deficits in obsessive-compulsive disorder (OCD). We tested the hypothesis that experimentally induced increments of subjective responsibility would lead to a disproportionately strong decline of memory confidence and enhanced response latencies in OCD while leaving memory accuracy unaffected. Twenty-eight OCD patients and 28 healthy controls were presented a computerized memory test framed with two different scenarios. In the neutral scenario, the participant was requested to imagine purchasing 15 items from a do-it-yourself store. In the recognition phase, the 15 needed items were presented along with 15 distractor items. The participant was asked to decide whether items were on his or her shopping list or not, graded by subjective confidence. In the responsibility scenario, the general experimental setup was analogous except that the participant now had to envision that he or she was a helper in a region recently struck by an earthquake, dispatched to provide 15 urgently needed goods from a nearby town. In line with prior work by our group, samples did not differ in either condition on memory accuracy in a subsequent recognition task. As hypothesized, OCD participants were less certain in their responses for the high responsibility condition than controls. Whereas patients and controls did not differ in their subjective estimates for memorized items, patients expressed stronger doubt that their earthquake mission was successful. The findings indicate that low memory confidence in OCD may only be elicited in situations where perceived responsibility is high and that patients may share higher performance standards ("good is not good enough") than controls when perceived responsibility is inflated. PMID- 17462590 TI - Skin conductance responses as predictor of emotional responses to stressful life events. AB - Preferential preattentive processing towards threat has been proposed to play a casual role in the development of anxiety, mainly because it reliably predicts emotional responding to stressful events. However, in the investigation of the predictive value of preferential preattentive processing towards threat, the emotional Stroop task has exclusively being used. This study was designed to prospectively investigate the predictive value of skin conductance reactivity in response to masked (i.e., preattentively perceived) threatening pictures on emotional responding following stressful life events. To collect skin conductance data, the picture perception task was administered to 136 police recruits due to enroll into active duty within 2-4 months. To assess psychological distress following emotionally stressful life events, the impact of event scale were administered approximately 24 months later. Controlling for trait anxiety, analyses indicated that enhanced skin conductance reactivity in response to masked threatening pictures (relative to neutrals) was a strong predictor of emotional responding to stressful life events. This finding support preferential preattentive processing towards threat as a relatively independent factor predictive of emotional responding. PMID- 17462591 TI - Characterization of vacuolar-ATPase and selective inhibition of vacuolar-H(+) ATPase in osteoclasts. AB - V-ATPase plays important roles in controlling the extra- and intra-cellular pH in eukaryotic cell, which is most crucial for cellular processes. V-ATPases are composed of a peripheral V(1) domain responsible for ATP hydrolysis and integral V(0) domain responsible for proton translocation. Osteoclasts are multinucleated cells responsible for bone resorption and relate to many common lytic bone disorders such as osteoporosis, bone aseptic loosening, and tumor-induced bone loss. This review summarizes the structure and function of V-ATPase and its subunit, the role of V-ATPase subunits in osteoclast function, V-ATPase inhibitors for osteoclast function, and highlights the importance of V-ATPase as a potential prime target for anti-resorptive agents. PMID- 17462592 TI - Discrete functions of rictor and raptor in cell growth regulation in Drosophila. AB - The TOR signaling pathway regulates cell growth and metabolism in response to various nutrient signals by forming complexes with either rictor or raptor. To distinguish the physiological roles of the complexes formed by the two different TOR partners, we compared the in vivo functions of rictor and raptor in Drosophila. In rictor-null mutants, Akt-induced tissue hyperplasia was reduced and Akt-Ser-505 phosphorylation was decreased. Furthermore, FOXO-dependent apoptosis, which is inhibited by Akt, was augmented in the rictor-null background, indicating that rictor is essential for the Akt-FOXO signaling module. We found that neither S6K-dependent cell growth nor S6K-Thr-398 phosphorylation was affected in rictor-null mutants. However, the knockdown of another TOR binding partner, raptor, decreased S6K-Thr-398 phosphorylation and inhibited S6K-induced cell overgrowth. Collectively, our findings strongly support that the association of TOR with rictor or raptor plays pivotal roles in TOR-mediated cell apoptosis and growth control by differentially regulating Akt- and S6K-dependent signaling pathways, respectively. PMID- 17462593 TI - Effects of VDAC isoforms on CuZn-superoxide dismutase activity in the intermembrane space of Saccharomyces cerevisiae mitochondria. AB - Copper and zinc containing superoxide dismutase (CuZnSOD) is located primarily in the cytosol but a small amount of the enzyme has also been identified in the intermembrane space of mitochondria (termed here IMS CuZnSOD). Using Saccharomyces cerevisiae mutants depleted of either isoform of VDAC (voltage dependent anion-selective channel), we have shown that the activity of IMS CuZnSOD coincides with the presence of a given VDAC isoform and changes in a growth phase dependent way. Moreover, the IMS CuZnSOD activity correlates with the levels of O2*- release from mitochondria and the cytosol redox state. The latter in turn seems to influence the levels of the mitochondrial outer membrane channel protein other than VDAC. Thus, we conclude that in the case of S. cerevisiae both VDAC isoforms influence the IMS CuZnSOD activity and subsequently the expression levels of some mitochondrial proteins. PMID- 17462594 TI - Functional interaction between dopamine receptor subtypes for the regulation of c fos expression. AB - Dopaminergic drugs increase the expression of the proto-oncogene, c-fos, in the brain, which is involved in the coordination of neurobiological changes caused by repeated cocaine or amphetamine use. This study examined the roles of five dopamine receptor subtypes on the c-fos promoter activity. D(1)R or D(5)R significantly increased the expression of c-fos promoter by activating protein kinase A. However, D(2)R, D(3)R, or D(4)R did not show any noticeable effects. The co-expression of D(1)R/D(3)R or D(1)R/D(2)R synergistically activated the basal and agonist-induced expression of the c-fos promoter, respectively. The Ral guanine-nucleotide-dissociation-stimulator-like, which was found to interact with the 3rd cytoplasmic loop of D(3)R, mediated the inhibitory activity of D(3)R in c fos expression. In summary, the expression of the c-fos promoter was increased by the D1-like receptors and enhanced synergistically by the D2-like receptors via the modulation of cellular cAMP. D(3)R inhibited the expression of the c-fos promoter through an interaction with RGL. PMID- 17462595 TI - Endocardiogenesis in embryoid bodies: novel markers identified by gene expression profiling. AB - Endocardial cells and cardiomyocytes differentiate from the cardiogenic mesoderm at about the same time during development. Although in vitro embryonic stem (ES) cell systems have been used to study the differentiation of various types of cell lineages, including cardiomyocytes, smooth muscle cells, and vascular endothelial cells, differentiation of endocardial cells, or endocardiogenesis, has not been well reported, because of a lack of specific molecular markers. In our search for cardiogenesis-associated genes expressed in embryoid bodies, we found several genes expressed in the heart region of mouse embryos, but not in cardiomyocytes. To identify the cell types expressing these genes, CD31(+) cells were taken from mouse embryos on embryonic day (E)8.5 and E9.5 and sorted, then their transcripts were analyzed using quantitative RT-PCR analyses. In those embryos, Gata4 and Nfatc1, as well as newly identified Cgnl1 and Dok4 were found to be preferentially expressed in endocardial cells, but not in yolk sac endothelial cells, while Cdh5 and Kdr were expressed in both cardiac and yolk sac endothelial cells. Immunohistochemical analyses of embryoid bodies revealed that some CD31(+) cells co-expressing Gata4 and Nfatc1 were located in close proximity to cardiomyocytes. These results suggest that embryoid bodies express endocardial specific genes and likely generate endocardial cells along with cardiomyocytes. Further, they indicate that these new marker genes are useful to study the origin and induction of endocardial cells, and identify other endocardial markers. PMID- 17462596 TI - Megalin-mediated endocytosis of cystatin C in proximal tubule cells. AB - Serum levels of cystatin C, an endogenous cysteine proteinase inhibitor, are often used as an indicator of glomerular filtration rate. Although it is known that cystatin C is filtered by glomeruli and metabolized in proximal tubule cells (PTC), the precise molecular mechanism underlying this process is undetermined. Using quartz-crystal microbalance analyses, we demonstrate that cystatin C binds directly to megalin, an endocytic receptor in PTC, in a Ca(+)-dependent manner. We also find that cystatin C is endocytosed specifically via megalin in rat yolk sac epithelium-derived L2 cells which share a variety of characteristics with PTC. Finally, in vivo studies using kidney-specific megalin knockout mice provide evidence that megalin mediates proximal tubular uptake of cystatin C. We conclude that megalin is an endocytic receptor of cystatin C in PTC. PMID- 17462597 TI - Human CD4+CD25+ regulatory T cells inhibit the differentiation of osteoclasts from peripheral blood mononuclear cells. AB - Regulatory T cell (Treg) is a subset of CD4+ T lymphocytes expressing CD25 with immunosuppressive activity. However the function of Tregs onto osteoclastogenesis remains unknown. We investigated the effect and regulatory mechanism of Treg focusing on osteoclastogenesis from PBMCs. Tregs were isolated from PBMCs by magnetic cell sorting-column and analyzed by flow cytometry. RT-PCR was performed to identify Foxp3 mRNA. Using PBMCs and Tregs coculture system, we could find that Tregs inhibited osteoclasts differentiation from PBMCs and reduced the resorbed areas on pit assay (p <0.01). This suppression of osteoclast differentiation was cytokine-dependent, not cell-to-cell direct contact proved by Transwell system. Tregs-induced osteoclast differentiation was blocked by anti TGF-beta or anti-IL-4 antibody treatment. These results suggest that Tregs inhibit osteoclast differentiation from PBMCs in a cytokine-dependent manner, not by cell-to-cell contact manner and that TGF-beta and IL-4 may be the key cytokines for this suppressive function of Tregs. PMID- 17462598 TI - Glucagon receptor expression and glucagon stimulation of ghrelin secretion in rat stomach. AB - The present study was performed to evaluate the role of glucagon in the regulation of ghrelin secretion from the rat stomach. mRNA for ghrelin and glucagon receptor was expressed predominantly in the lower body and pylorus of stomach, but little or not in the upper body and cardia. Ghrelin- and glucagon receptor-immunoreactive cells were detected in lamina propria mucosae of stomach and some cells expressed both. Intravenous administration of glucagon caused transient increases in both acyl- and desacyl-ghrelin levels in the gastric vein within 10 min, which was followed by gradual increases in desacyl-ghrelin levels until 60 min. Steady state levels of ghrelin mRNA in the stomach were increased by 1.9-fold 20 min after glucagon administration, but not at 5 or 120 min. These results suggest that glucagon stimulates acute release of both forms of ghrelin and thereafter upregulates synthesis and release of desacyl-ghrelin in the rat stomach. PMID- 17462599 TI - A novel highly acidic polysaccharide with inhibitory activity on calcification from the calcified scale "coccolith" of a coccolithophorid alga, Pleurochrysis haptonemofera. AB - Coccolith, a calcified scale with species-specific fine structure produced by marine unicellular coccolithophorid algae, consists of calcium carbonate (CaCO(3)) crystals and a small amount of organic matrices. A novel polysaccharide named coccolith matrix acidic polysaccharide (CMAP) was isolated from the coccolith of a coccolithophorid alga, Pleurochrysis haptonemofera. The structure of CMAP was determined by chemical analysis and NMR spectroscopy including COSY, TOCSY, HMQC, and HMBC to be a polysaccharide composed of the following unit: - >4) l-iduronic acid (alpha1-->2) meso-tartaric acid (3-->1) glyoxylic acid (1-->. It has four carboxyl groups per a disaccharide unit as observed in another polysaccharide PS-2 characterized previously in Pleurochrysis carterae. CMAP showed a strong inhibitory activity on CaCO(3) precipitation. These results suggest that CMAP serves as a regulator in the calcification of the coccolith. PMID- 17462600 TI - Involvement of Rabring7 in EGF receptor degradation as an E3 ligase. AB - Rab7, a member of the Rab family small G proteins, is involved in the late stage of the endocytic pathway. We previously identified a Rab7 target protein, Rabring7, which contains a RING finger domain at its C termini, but the precise role of Rabring7 remains unknown. In this study, we demonstrate using an in vitro ubiquitination assay with recombinant E1 and E2 proteins that Rabring7 has E3 ligase activity and that it preferentially reacts with Ubc4 and Ubc5 as its E2 proteins. Rabring7 ubiquitinated itself but not Rab7, and a mutation at Cys-229 in the RING finger domain (Rabring7C229S) completely diminished its E3 ligase activity. In the ligand-induced degradation of EGF receptor (EGFR), Rabring7 accelerated the degradation of EGFR, whereas Rabring7C229S inhibited the degradation induced by cCbl, another E3 ligase. These results suggest that Rabring7 is involved in the endocytic trafficking of EGFR through its E3 ligase activity. PMID- 17462601 TI - An antibody directed against PDGF receptor beta enhances the antitumor and the anti-angiogenic activities of an anti-VEGF receptor 2 antibody. AB - Platelet-derived growth factor (PDGF) and its receptors (PDGFR) play important roles in tumorigenesis through stimulating tumor growth and promoting angiogenesis via enhancing pericyte recruitment and vessel maturation. Here we produced a neutralizing antibody, 1B3, directed against mouse PDGFRbeta. 1B3 binds to PDGFRbeta with high affinity (9x10(-11)M) and blocks PDGF-BB from binding to the receptor with an IC(50) of approximately 1.2 nM. The antibody also blocks ligand-stimulated activation of PDGFRbeta and downstream signaling molecules, including Akt and MAPK p42/44, in tumor cells. In animal studies, 1B3 significantly enhanced the antitumor and the anti-angiogenic activities of DC101, an antibody directed against mouse vascular endothelial growth factor receptor 2, in a pancreatic (BxPC-3) and a non-small cell lung (NCI-H460) tumor xenograft models. Treatment with the combination of 1B3 and DC101 in BxPC-3 xenograft bearing mice resulted in tumor regression in 58% of mice compared to that in 18% of mice treated with DC101 alone. Taken together, these results lend great support to use PDGFRbeta antagonists in combinations with other antitumor and/or anti-angiogenic agents in the treatment of a variety of cancers. PMID- 17462602 TI - A novel peptide-nucleotide dual vaccine of human telomerase reverse transcriptase induces a potent cytotoxic T-cell response in vivo. AB - Human telomerase reverse transcriptase (hTERT) is highly expressed in over 85% of human cancers, which makes it a broadly applicable molecular target for cancer therapy. Several groups have demonstrated that hTERT can efficiently evoke specific cytotoxic T lymphocytes (CTL) responses for malignant tumors. In the present study, we developed a novel virus-like particulate peptide-nucleotide dual vaccine (PNDV) of hTERT, which was composed of a low-affinity epitope variant with encoding full-length gene in the same virus-size particulate. We verified the formation of PNDV by DNA retarding assay, DNase I protection assay and transmission electron microscopy, and confirmed its immunogenicity and transfection activities in mammalian cells. Furthermore, in vivo immunization of HLA-A2.1 transgenic mice generated efficient IFN-gamma secretion and hTERT specific CTLs which are known to cause selective cell death of telomerase positive gastrointestinal cancer cells. To our knowledge, this represents the first report on collocating a low-affinity epitope variant with a full-length hTERT gene for anti-cancer vaccine design. This novel strategy for vaccine design not only enables enhanced immunity to a universal tumor antigen, but also has the potential to generate CTLs effective in telomerase-positive tumor cells of diverse tissue origins. Therefore, our findings bear significant implications for immunotherapy of human cancers. PMID- 17462603 TI - Wnt3a binds to several sFRPs in the nanomolar range. AB - Secreted Frizzled-related proteins (sFRPs) are modulators of the Wnt signaling pathway that plays important roles in both embryogenesis and oncogenesis. sFRPs have been proposed to antagonize Wnt activity by binding to Wnts. However, the affinity of this binding is unknown. Here we show, using surface plasmon resonance and purified proteins, that sFRP1, sFRP2, sFRP4, and Frzb bind directly to Wnt3a with affinities in the nanomolar range. However, only sFRP1 and sFRP2 antagonize Wnt3a activity by blocking Wnt3a induced beta-catenin accumulation in L cells. Furthermore, sFRP2, but not Frzb, antagonizes Wnt3a signaling in an ES cell model of mesoderm differentiation. These results provide the first measurement of binding affinity of sFRPs for a Wnt, which together with the measurement of antagonistic activity of sFRPs could help understand how sFRPs regulate Wnt signaling. PMID- 17462604 TI - Interaction and transport characteristics of mycophenolic acid and its glucuronide via human organic anion transporters hOAT1 and hOAT3. AB - The immunosuppressant mycophenolate mofetil (MMF) is frequently administered with calcineurin inhibitors and corticosteroids to recipients of organ transplantations. However, the renal handling of the active metabolite mycophenolic acid (MPA) and 7-O-MPA-glucuronide (MPAG) has been unclear. The purpose of the present study was to assess the interaction of MPA and MPAG with the human renal organic anion transporters hOAT1 (SLC22A6) and hOAT3 (SLC22A8), by conducting uptake experiments using HEK293 cells stably expressing these transporters. MPA and MPAG inhibited the time-dependent uptake of p [(14)C]aminohippurate by hOAT1 and that of [(3)H]estrone sulfate by hOAT3. The apparent 50% inhibitory concentration (IC(50)) of MPA for hOAT1 and hOAT3 was estimated at 10.7 and 1.5 microM, respectively. In the case of MPAG, the IC(50) values were calculated at 512.3 microM for hOAT1 and 69.1 microM for hOAT3. Eadie Hofstee plot analyses showed that they inhibited hOAT1 noncompetitively and hOAT3 competitively. No inhibitory effects of tacrolimus, cyclosporin A and azathioprine on transport of p-[(14)C]aminohippurate by hOAT1 and of [(3)H]estrone sulfate by hOAT3 were observed. No transport of MPA by these transporters was observed. On the other hand, the uptake of MPAG into cells was stimulated by the expression of hOAT3, but not hOAT1. These findings propose the possibility that the administration of MMF decreases the renal clearance of drugs which are substrates of hOAT1 and hOAT3. Present data suggest that hOAT3 contributes to the renal tubular secretion of MPAG. PMID- 17462605 TI - Activity of "reversed" diamidines against Trypanosoma cruzi "in vitro". AB - Chagas' disease is an important parasitic illness caused by the flagellated protozoan Trypanosoma cruzi. The disease affects nearly 17 million individuals in endemic areas of Latin America and the current chemotherapy is quite unsatisfactory based on nitroheterocyclic agents (nifurtimox and benznidazol). The need for new compounds with different modes of action is clear. Due to the broad-spectrum antimicrobial activity of the aromatic dicationic compounds, this study focused on the activity of four such diamidines (DB811, DB889, DB786, DB702) and a closely related diguanidine (DB711) against bloodstream trypomastigotes as well as intracellular amastigotes of T. cruzi in vitro. Additional studies were also conducted to access the toxicity of the compounds against mammalian cells in vitro. Our data show that the four diamidines compounds presented early and high anti-parasitic activity (IC50 in low micromolecular range) exhibiting trypanocidal dose-dependent effects against both trypomastigote and amastigote forms of T. cruzi 2h after drug treatment. Most of the diamidines compounds (except the DB702) exerted high anti-parasitic activity and low toxicity to the mammalian cells. Our results show the activity of reversed diamidines against T. cruzi and suggested that the compounds merit in vivo studies. PMID- 17462606 TI - Kinetics of glyburide metabolism by hepatic and placental microsomes of human and baboon. AB - Glyburide (glibenclamide) is under investigation for treatment of gestational diabetes. Two metabolites of glyburide have been previously identified in patients, namely, 4-trans-(M1) and 3-cis-(M2) hydroxycyclohexyl glyburide. Recently, the metabolism of glyburide by microsomes of liver and placenta from humans and baboons revealed the formation of four additional metabolites: 4-cis (M2a), 3-trans-(M3), and 2-trans-(M4) hydroxycyclohexyl glyburide, and ethyl hydroxy glyburide (M5). The aim of this investigation was to determine the kinetics for the metabolism of glyburide by cytochrome P450 (CYP) isozymes of human and baboon placental and hepatic microsomes. The metabolism of glyburide by microsomes from the four organs revealed saturation kinetics and apparent K(m) values between 4 and 12 microM. However, the rates for formation of the metabolites varied between organs and species. M1 was the major metabolite (36% of total), formed by human hepatic microsomes with V(max) of 80+/-13 pmol mg protein(-1)min(-1), and together with M2, accounted for only 51% of the total. M5 was the major metabolite (87%) formed by human placental microsomes with V(max) of 11 pmol mg protein(-1)min(-1). In baboon liver, M5 had the highest rate of formation (V(max) 135+/-32 pmol mg protein(-1)min(-1), 39% of total), and in its placenta, was M4 (V(max) 0.7+/-0.1 pmol mg protein(-1)min(-1), 65%). The activity of human and baboon hepatic microsomes in metabolizing glyburide was similar, but the activity of human and baboon placental microsomes was 7% and 0.3% of their respective hepatic microsomes. The data obtained suggest that more than 1 CYP isozyme is responsible for catalyzing the hydroxylation of glyburide. PMID- 17462607 TI - Failure of estradiol to ameliorate global ischemia-induced CA1 sector injury in middle-aged female gerbils. AB - Global forebrain ischemia arising from brief occlusion of the carotid arteries in gerbils produces selective hippocampal CA1 neuronal loss. Pre-treatment with 17beta-estradiol ameliorates, in part, ischemia-induced damage in young animals. Because stroke and cardiac arrest are more likely to occur among elderly individuals, neuroprotective studies in older animals have compelling clinical relevance. We investigated whether estradiol would attenuate ischemia-induced hippocampal neuronal injury in middle-aged (12-14 months) male, intact female, ovariectomized (OVX) female and OVX females treated for 14 days with estradiol. Core temperature telemetry probes were also implanted at the time that estradiol was initiated. Ischemia was induced by bilateral occlusion of the common carotid arteries (5 min), during which time skull temperature was maintained under normothermic conditions. Estradiol blocked the modest spontaneous hyperthermia that normally follows ischemia. However, all four groups exhibited substantial neuronal cell loss in the CA1, assessed at 7 after ischemia. These findings indicate that estradiol pre-treatment under conditions that produce neuroprotection in young animals does not protect against ischemia-induced CA1 cell loss in middle-aged female gerbils. PMID- 17462608 TI - Therapeutic effects of hyperbaric oxygen in a rat model of endothelin-1-induced focal cerebral ischemia. AB - It has been established that hyperbaric oxygen (HBO) treatment reduces brain edema, decreases infarct volume, contributes to neurological functional recovery and suppresses apoptosis in suture-induced focal cerebral ischemic animal models. In the present study, we evaluated the therapeutic effect of HBO in an endothelin 1-induced focal cerebral ischemia in rats and explored the associated mechanisms of HBO-induced brain protection. One hundred twenty male Sprague-Dawley rats (280 to 320 g) were randomly assigned to sham, focal cerebral ischemia and focal cerebral ischemia treated with HBO groups. Brain water content, neurological function, morphology and molecular biological markers were assessed. HBO (100% O2, 2.5 atmosphere absolute for 2 h) was initiated at 1 h after focal cerebral ischemia. Rats were killed at 24 h to harvest tissues for Western blot or for histology. In HBO-treated animals, an enhanced ratio of Bcl-2 and Bax and a reduced expression of hypoxia-inducible factor-1alpha (HIF-1alpha) in the hippocampus after focal cerebral ischemia were observed. These results indicate that HBO provides brain protection that is probably associated with the inhibition of HIF-1alpha and the elevation of Bcl-2. PMID- 17462609 TI - Secretory PLA2-IIA and ROS generation in peripheral mitochondria are critical for neuronal death. AB - In this study the role of mitochondrial secretory PLA2-IIA in glutamate-induced cell death in cultured cerebellar granule neurons has been investigated. Inhibition of secretory PLA2-IIA blocked glutamate-induced cell death. Since PLA2 may generate reactive oxygen species (ROS), we have investigated ROS production, detected as dihydrorhodamine 123 oxidation and nitrotyrosine modifications of proteins, following glutamate treatment in the absence or presence of an inhibitor of secretory PLA2-IIA. There was an increased generation of ROS in both glutamate- and buffer-treated neurons compared to untreated neurons. Scavenging with dihydrorhodamine 123 reduced glutamate-induced death (60%), showing that ROS detected in glutamate-treated neurons were associated with cell death. However, ROS detected in buffer-treated neurons were not associated with toxicity. Glutamate treatment led to ROS production predominantly in peripheral mitochondria, whereas buffer treatment led to ROS production in somal mitochondria. Inhibition of secretory PLA2-IIA (i) reduced the generation of ROS after glutamate treatment, (ii) reduced the ROS production in peripheral mitochondria in glutamate-treated neurons, consistent with the fact that calcium entry through glutamate (NMDA) receptors has a privileged access to peripheral mitochondria, and (iii) did not reduce the generation of ROS after buffer treatment. In conclusion, activation of NMDA receptors induces ROS, which is critical for neuronal death, due to secretory PLA2-IIA associated with peripheral mitochondria. PMID- 17462610 TI - Seeing yourself in a positive light: brain correlates of the self-positivity bias. AB - Individuals are found to have better recall for self-referent information than other types of information. However, attribution research has shown that self reference is highly correlated with emotional valence. The present study attempted to identify and separate the processing of self-reference and emotional valence using ERPs. Participants performed a two-choice task, judging the self referential content of positive and negative words. Reaction times revealed an interaction between self-reference and emotional valence. Faster responses occurred after self-positive and non-self negative words as compared to self negative and non-self-positive words. A similar interaction was identified in ERP waveforms in the time range of the N400 component at fronto-central electrode sites, with larger N400 amplitudes for words outwith the self-positivity bias. Thus, the size of the N400 may indicate the extent to which information is discrepant with the individual's self-concept. PMID- 17462611 TI - Regulation of the Na+/Ca2+ exchanger (NCX) in the murine embryonic heart. AB - OBJECTIVE: The Na+/Ca2+ exchanger (NCX) is involved in embryonic heart development and function demonstrated by the abnormal myofibrillar organization, defects in heartbeat, and early embryonic death of NCX-null embryos. It was therefore the aim of our study to identify key functional regulators of the embryonic NCX. METHODS: NCX current (I(NCX)) density was measured as the Ni2+ (5 mM)-sensitive current applying the whole-cell patch-clamp technique in early (EDS, E10.5V) and late developmental stage (LDS, E16.5V) mouse ventricular cardiomyocytes. RESULTS: Compared to LDS, cardiomyocytes derived from EDS showed a significantly higher basal I(NCX) density for the I(NCX) forward (-120 mV: 4.1+/-1 pA/pF, n=15 versus -1.7+/-0.4, n=11, p<0.05) and reverse modes (+60 mV: 4.0+/-0.9 pA/pF, n=15 versus 1.8+/-0.4, n=11, p<0.05). There was 2-3-fold elevation of forward and reverse current in LDS on application of ATP-gamma-S (2 mM) together with forskolin (1 microM) as well as intracellular application of the catalytic subunit of cAMP-dependent protein kinase (cPKA, 200 U/mL), cAMP (200 microM), phorbol 12-myristate-13-acetate (PMA), a direct activator of protein kinase C (PKC), and 8-Br-cGMP, a membrane permeable analog of cGMP. The specific PKC inhibitor Ro 31-8220 significantly reduced I(NCX) by 70%. Co application of 20 microM PKA inhibitor Fragment 14-22 (PKI), a specific inhibitor of PKA, and cAMP significantly reduced the exchanger activity by approx 60%. Despite these obvious effects in LDS we could not detect a significant impact of these compounds on I(NCX) in EDS-derived cardiomyocytes. Application of the alkaline phosphatase to test for constitutive phosphorylation of NCX did not affect I(NCX) density in LDS but led to an approx 80% reduction of I(NCX) in EDS. CONCLUSION: In EDS cardiomyocytes I(NCX) density is upregulated, at least in part by the high phosphorylation of the exchanger protein. At LDS, embryonic cardiomyocytes showed a strong increase of I(NCX) density upon stimulation by PKC and PKA-dependent signalling pathways. PMID- 17462612 TI - Validation of a quality assessment system for blood gas and electrolyte testing. AB - BACKGROUND: A recently-introduced quality assessment system (Intelligent Quality Management: iQM), was evaluated in routine clinical use at four different hospitals. The iQM technology is designed to replace conventional external liquid controls with software, Process Control (PC) Solutions and Calibration Validation components that continually assess the function of the GEM Premier 3000 (GEM) analyzer and automatically initiate and document corrective actions. METHODS: We validated the performance claims of iQM by monitoring quality control (QC) materials at 4 clinical sites while analyzing approximately 10,550 patient samples. We compared iQM-measured QC values to traditional QC results, evaluating the number and type of error flags for patient samples, and used data from control results to calculate the average time to detect an error (ADT) for each analyte. RESULTS: The calculated ADT was approximately 3 min for all analytes except for sodium (17 min), glucose (11 min), and lactate (5.9 min). Precision of control materials in iQM cartridges was better than from external controls run on traditional analyzers. The iQM system detected errors in 0.46% of actual clinical samples. CONCLUSIONS: The findings from our study confirm that (a) iQM precision in a clinical setting is comparable to that found in previous studies done in a research setting, (b) the improved precision of control material on the iQM is likely because the internal control fluids are sealed and not susceptible to exposure from handling, and (c) the system detects and often corrects errors in specific samples that might not be reported by traditional analytical systems. PMID- 17462613 TI - Immunoreactive circulating oxidized HDL concentrations do not increase in patients undergoing carotid endarterectomy: a comparative study for oxidized HDL and oxidized LDL concentrations in plasma. PMID- 17462614 TI - Column-switching LC-MS/MS analysis for quantitative determination of testosterone in human serum. AB - BACKGROUND: An accurate measurement of testosterone is needed in many clinical applications for correct diagnosis and appropriate treatment. Our aim was to develop a fast and robust high-throughput LC-MSMS method for quantification of serum testosterone in women. METHODS: Testosterone was derivatized by oximation and extracted with methyl tert-butyl ether from 200 microL of serum. Further matrix elimination was achieved on-line using a column-switching LC-method. The instrumental analysis was performed on an API4000 tandem mass spectrometer equipped with an Agilent series 1312A binary pump and an Agilent series 1311A quaternary pump. The MRM transitions were 304-->124 and 304-->112 for testosterone and 307-->124 and 307-->112 for d(3)-testosterone. RESULTS: The total analysis time of the column-switching method was 3 min. Linear calibration curves were obtained in the concentration range from 0.035 nmol/L (0.01 microg/L) to 6.92 nmol/L (2 microg/L). Within-day and between-day precision, expressed as the relative standard deviation at four different concentrations ranged from 4.70% to 9.35%. Correlation with the in-house method (solvent-extraction RIA) showed r(2)=0.920. CONCLUSIONS: The presented column-switching method offers a simple, fast and economical analysis of testosterone in human serum. The procedure requires only small sample volumes and is well suited for quantification of testosterone in serum from women and children. PMID- 17462615 TI - Simultaneous determination and pharmacokinetic study of roxithromycin and ambroxol hydrochloride in human plasma by LC-MS/MS. AB - BACKGROUND: Although roxithromycin and ambroxol HCl were often administered concomitantly for the treatment of respiratory infections, the pharmacokinetic interactions between them have not been reported. We investigated the interactions between these drugs in health male Chinese volunteers by LC-MS/MS in human plasma. METHODS: The pharmacokinetics were studied in 12 healthy male Chinese volunteers after an overnight fast by a single oral dose, 4-way crossover design with a period of 7-day washout. Each subjects was randomized to receive a single oral dose of 1 compound roxithromycin (150 mg) and ambroxol HCl (30 mg) dispersible tablet (test formulation, treatment A), one 150 mg roxithromycin dispersible tablet together with one 30 mg ambroxol HCl tablet (combined reference formulations, treatment B), one 150 mg roxithromycin dispersible tablet (reference formulation I, treatment C), or one 30 mg ambroxol HCl tablet (reference formulation II, treatment D) with 250 ml of water. Venous blood was collected at pre-dose (0 h) and 0.25, 0.5, 1, 1.5, 2, 2.5, 3, 4, 6, 8, 12, 24, 48, 72 h after dosing. The plasma concentrations of roxithromycin and ambroxol HCl were simultaneously determined by using a validated internal standard LC MS/MS method. RESULTS: No significant differences were observed for the major pharmacokinetic parameters such as C(max), T(max), t(1/2) and AUC of both roxithromycin and ambroxol HCl between different treatments. CONCLUSION: The pharmacokinetics of both roxithromycin and ambroxol HCl are not affected by their concomitant oral administration. Therefore, there are no obvious pharmacokinetic interactions between roxithromycin and ambroxol HCl after oral administration. Roxithromycin and ambroxol HCl dispersible tablets were bioequivalent with reference to the roxithromycin dispersible tablets and ambroxol HCl tablets in combination usage. PMID- 17462616 TI - Mutual interference between serum thyroglobulin and antithyroglobulin antibody in an automated chemiluminescent immunoassay. AB - OBJECTIVES: To investigate the analytical interference between serum Tg and TgAb. DESIGN AND METHODS: Tg and TgAb were measured on an automated chemiluminescent immunoassay system in mixed sera from DTC patients and individual samples spiked with exogenous Tg. RESULTS: Tg and TgAb recoveries in mixed patient samples were inversely correlated with expected TgAb or Tg concentrations, respectively. Impaired TgAb recoveries were also found in 50% (10/20) samples with high Tg in the exogenous recovery tests. CONCLUSIONS: Mutual but not equal analytical interference between Tg and TgAb is present and concentration-dependent with interpatient variability. PMID- 17462617 TI - Human erythrocyte delta-aminolevulinate dehydratase activity and oxidative stress in hemodialysis patients. AB - BACKGROUND: Oxidative stress is a complicating factor in chronic renal failure, especially in hemodialysis (HD) patients. Also, aluminum intoxication may occur during hemodialysis treatment. Aluminum has been shown to inhibit the sulfhydryl containing enzyme delta-aminolevulinate dehydratase (ALA-D). Thus, the involvement of -SH oxidation in ALA-D inhibition and its relationship with serum Al levels and lipid peroxidation in HD patients were evaluated. METHODS: Blood ALA-D activity, plasma thiobarbituric acid reactive substances (TBARS), and serum aluminum levels were measured in HD patients (n=37) and healthy controls (n=20). RESULTS: TBARS and Al levels were higher in HD patients than in controls (p<0.01), while ALA-D activity was lower (p<0.05). The sulfhydryl-reducing agent dithiothreitol (DTT) reactivated ALA-D of HD patients, but activity was still lower than that of controls. ALA-D activity was negatively correlated with TBARS (r=-0.63, p<0.01) and aluminum levels (r=-0.31, p<0.05). CONCLUSIONS: Reduced ALA D activity in HD patients was found to be related to the oxidation of -SH groups essential for enzyme activity. Our results suggest that increased oxidative stress may have contributed to enzyme inhibition in HD patients. PMID- 17462618 TI - Comparative analysis of zinc protoporphyrin and blood lead levels in lead-exposed Andean children. AB - OBJECTIVES: The objective of this study was to investigate the zinc protoporphyrin/heme ratio (ZPP/heme) as a biomarker for chronic lead (Pb) poisoning in children with a history of high Pb exposure. DESIGN AND METHODS: ZPP/heme ratio was measured in blood samples from 78 children (44 females and 34 males) with persistent Pb exposure from Pb glazing of ceramics in a local cottage industry in the Andes Mountains of Ecuador. RESULTS: Mean blood lead (PbB) level was 26.4 microg/dL (SD: 23.2; range: 4.0-107.0), and the mean ZPP/heme ratio was 152.4 micromol/mol (SD: 190.6; range: 36.0-1064.0). A regression analysis of PbB level and ZPP/heme ratio revealed a significant association (r=0.761, p= <0.0001), with the logZPP showing a higher correlation with PbB (r=0.869, p= <0.0001). The ZZP/heme ratio decreased significantly with increasing age (ANOVA, p=0.030). The mean ZPP/heme ratios for females and males were 139.6 and 169.0 micromol/mol, respectively, and were not statistically different (t-test, p=0.504). CONCLUSION: The elevated ZPP/heme ratios, coupled with high PbB levels observed in this cohort of Andean children of Pb-glazing workers, suggest chronic Pb intoxication and probable iron deficiency. PMID- 17462619 TI - Variability in apo(a) gene regulatory sequences, compound genotypes, and association with Lp(a) plasma levels. AB - OBJECTIVES: Lipoprotein(a) is an independent risk factor of atherosclerosis. DESIGN AND METHODS: We assigned frequencies of six polymorphic sites from apo(a) gene transcription control regions, linkage disequilibrium, and 5-polymorphic compound genotypes association with Lp(a) levels. RESULTS: Significant linkage disequilibrium between polymorphic sites was detected. Compound genotypes were significantly associated with Lp(a) levels (P<0.0001). Major 5-polymorphic genotypes were distributed in a broad range of concentrations. CONCLUSIONS: Major 5-polymorphic compound genotypes are not associated with restricted range of Lp(a) levels. PMID- 17462620 TI - Representational change and children's numerical estimation. AB - We applied overlapping waves theory and microgenetic methods to examine how children improve their estimation proficiency, and in particular how they shift from reliance on immature to mature representations of numerical magnitude. We also tested the theoretical prediction that feedback on problems on which the discrepancy between two representations is greatest will cause the greatest representational change. Second graders who initially were assessed as relying on an immature representation were presented feedback that varied in degree of discrepancy between the predictions of the mature and immature representations. The most discrepant feedback produced the greatest representational change. The change was strikingly abrupt, often occurring after a single feedback trial, and impressively broad, affecting estimates over the entire range of numbers from 0 to 1000. The findings indicated that cognitive change can occur at the level of an entire representation, rather than always involving a sequence of local repairs. PMID- 17462621 TI - Ultrastructural observations of cryoinjury in kangaroo spermatozoa. AB - Macropod spermatozoa have proven difficult to cryopreserve such that empirical studies using high concentrations of glycerol and/or DSMO have resulted in only 10% post-thaw motility. We examined the ultrastructure and freeze-fracture of caput and cauda epididymal macropod spermatozoa at 35, 4 degrees C and following cryopreservation with and without 20% glycerol. The addition of 20% glycerol resulted in significant damage to the sperm plasma membrane and mitochondria compared to no glycerol at the same temperatures (P<0.05). Following cryopreservation, 20% glycerol significantly improved the preservation of the cauda epididymal sperm plasma membrane and mitochondria and reduced the incidence of axonemal damage and axonemal spaces. For caput epididymal spermatozoa, glycerol only improved the preservation of the plasma membrane following cryopreservation (P<0.05). Freeze fracture microscopy revealed a pattern of helically wound intramembranous particles in the plasma membrane over the fibre network of the mid piece of the sperm tail. The fibre network is an interconnecting cytoskeletal structure found underneath the plasma membrane of the kangaroo sperm midpiece and is thought to add rigidity to the proximal portion of the sperm tail. After thawing, the plasma membrane was damaged such that this structure was missing in patches, and the helical rows of particles were mal-aligned. On the principal piece, particles were arranged randomly at physiological temperatures; however, upon cooling to 4 degrees C with 20% glycerol, the particles become aggregated. Once rewarmed (35 degrees C), particles over the principal piece resumed their random organisation. This finding is further evidence of a reversible phase transition of the macropod sperm plasma membrane during cooling that is not associated with a loss of motility or membrane integrity. PMID- 17462622 TI - Divergent evolution of the vertebrate polysialyltransferase Stx and Pst genes revealed by fish-to-mammal comparison. AB - Polysialic acid (PSA) is a developmentally regulated carbohydrate attached to the neural cell adhesion molecule (NCAM). PSA is involved in dynamic processes like cell migration, neurite outgrowth and neuronal plasticity. In mammals, polysialylation of NCAM is catalyzed independently by two polysialyltransferases, STX (ST8Sia II) and PST (ST8Sia IV), with STX mainly acting during early development and PST at later stages and into adulthood. Here, we functionally characterize zebrafish Stx and Pst homolog genes during fish development and evaluate their catalytic affinity for NCAM in vitro. Both genes have the typical gene architecture and share conserved synteny with their mammalian homologues. Expression analysis, gene-targeted knockdown experiments and in vitro catalytic assays indicate that zebrafish Stx is the principal--if not unique- polysialyltransferase performing NCAM-PSA modifications in both developing and adult fish. The knockdown of Stx exclusively affects PSA synthesis, producing defects in axonal growth and guidance. Zebrafish Pst is in principle capable of synthesizing PSA, however, our data argue against a fundamental function of the enzyme during development. Our findings reveal an important divergence of Stx and Pst enzymes in vertebrates, which is also characterized by a differential gene loss and rapid evolution of Pst genes within the bony-fish class. PMID- 17462623 TI - Ambient particulate matter (PM2.5): physicochemical characterization and metabolic activation of the organic fraction in human lung epithelial cells (A549). AB - To contribute to complete the knowledge of the underlying mechanisms of action involved in air pollution particulate matter (PM)-induced cytotoxicity, an aerosol was collected in Dunkerque, a French seaside City heavily industrialized. In this work, we focused our attention on its physical and chemical characteristics, its cytotoxicity, and its role in the induction of the volatile organic compound (VOC) and/or polycyclic aromatic hydrocarbon (PAH)-metabolizing enzymes in human lung epithelial cells (A549). Size distribution showed that 92.15% of the collected PM were PM2.5 and the specific surface area was 1 m2/g. Inorganic (i.e. Fe, Al, Ca, Na, K, Mg, Pb, etc.) and organic (i.e. VOC, PAH, etc.) chemicals were found in collected PM, revealing that much of them derived from wind-borne dust from the industrial complex and the heavy motor vehicle traffic. The thermal desorption study indicated that organic chemicals were not only adsorbed onto the surface but also highly incrusted in the structure of PM. The lethal concentrations at 10% and 50% of collected PM were 23.72 microg/mL (or 6.33microg/cm2) and 118.60 microg/mL (or 31.63 microg/cm2), respectively. The VOC and/or PAH-coated onto PM induced significant increases in mRNA expressions of cytochrome P450 (cyp) 1a1, cyp2e1, cyp2f1, nadph quinone oxydo-reductase-1, and glutathione s-transferase-pi 1, versus controls. Hence, we concluded that the metabolic activation of the very low doses of VOC and/or PAH-coated onto the inorganic condensation nuclei from Dunkerque City's PM is one of the underlying mechanisms of action closely involved in its cytotoxicity in human lung epithelial cells. PMID- 17462624 TI - Effects of acute and chronic treatment with fluoxetine on stress-induced hyperthermia in telemetered rats and mice. AB - Preclinical and clinical evidence suggests that anxiolytic effects are observed after chronic administration of the selective serotonin reuptake inhibitor fluoxetine. In contrast, acute treatment may increase signs of anxiety. The present study examined the effects of acute and chronic administration of fluoxetine on a physiological measure of anxiety, stress-induced hyperthermia, in rats and mice using radiotelemetry to record core temperature and locomotor activity and ethologically relevant stressors to evoke the hyperthermic response. In both species, the benzodiazepine agonist chlordiazepoxide reduced stress induced hyperthermia at doses (5 mg/kg i.p. rat, 10 mg/kg p.o. mouse) that had no significant effect on locomotor activity. Similarly, in both species, chronic (21 days) treatment with fluoxetine attenuated the hyperthermic response without significantly affecting locomotor activity. However, acute fluoxetine elicited species-specific effects. Thus in mice, stress-induced hyperthermia and activity were unaffected by fluoxetine (20 mg/kg p.o.) consistent with a lack of anxiolytic or anxiogenic activity. In contrast, in rats, fluoxetine (10 mg/kg i.p.) caused a significant baseline hypothermia in the absence of stress, confounding further interpretation. In conclusion, stress-induced hyperthermia in mice was unaffected by acute treatment and significantly reduced by chronic treatment with fluoxetine. However, in rats chronic administration of fluoxetine significantly reduced stress-induced hyperthermia while the effects of acute treatment were confounded by a decrease in body temperature in the absence of stress. Together, these observations support the view that chronic administration of fluoxetine is anxiolytic; however, the stress-induced hyperthermia assay does not reveal anxiogenic effects of acute administration of fluoxetine in rats or mice. PMID- 17462625 TI - Anti-fibrotic effect of meloxicam in a murine lung fibrosis model. AB - A murine lung fibrosis model has been induced by challenging male Swiss albino mice with a fibrotic dose of bleomycin (10 mg/kg body weight, s.c.) twice weekly for 6 weeks. The model has been characterized and confirmed biochemically, histologically and morphometrically. Keeping in mind that inflammation is the forerunner of lung fibrosis, we have investigated the possible anti-fibrotic effect of meloxicam; a selective COX-2 inhibitor, in this lung fibrosis paradigm. When administered ahead of bleomycin challenge, meloxicam significantly reduced the lung content of hydroxyproline; the backbone of collagen matrix. This was further confirmed by the lower collagen deposition as revealed by histochemical examination of lung sections. Meloxicam had also anti-oxidant effect as shown by increase in lung reduced glutathione (GSH) level and decreases in lung malonedialdehyde (MDA) content and myeloperoxidase (MPO) activity. Besides, meloxicam has shown an apparent angiostatic activity. Histologically, meloxicam lessened lung inflammation and fibrotic changes induced by bleomycin. Taken together, one could conclude that meloxicam has shown anti-fibrotic effect in the bleomycin lung fibrosis model. Apart from its well-known anti-inflammatory potential, this anti-fibrotic action of meloxicam resides most probably, at least partly, in its anti-oxidant and angiostatic effects. PMID- 17462626 TI - Functional analysis of RalGPS2, a murine guanine nucleotide exchange factor for RalA GTPase. AB - RalGPS2 is a murine guanine nucleotide exchange factor of the RalGPS family; it contains a Cdc25-like GEF domain and does not exhibit a Ras-binding domain. The main characteristic of RalGPS2 is its pleckstrin homology (PH) domain, present at the C terminus, that preferentially binds phosphatidylinositol-4,5-biphosphate and in HEK 293 cells localized in membranes, causing ruffling and vesiculation. Moreover, RalGPS2 contains a PxxP motif in the central part of the molecule. This motif binds in vitro and in vivo SH3 domains of Grb2 and PLCgamma. RalGPS2 and its GEF domain activate RalA in vivo while the PH-PxxP domains inhibited it behaving as a dominant negative for the RalA pathway; this activation was not inhibited by co-expression of a dominant negative Ras. RalGPS2 is physiologically expressed in testis and brain; when overexpressed, the whole RalGPS2 causes considerable morphological changes in HEK 293 cells, suggesting its possible role on cytoskeleton reorganization. This is further strengthened by data obtained in NIH3T3 cells where expression of PH-PxxP domain promotes actin depolymerization. Finally, RalGPS2 and its GEF domain induce Ras-independent transcriptional activation of the c-fos promoter in NIH3T3 cells. PMID- 17462627 TI - Distinct functional domains in nesprin-1alpha and nesprin-2beta bind directly to emerin and both interactions are disrupted in X-linked Emery-Dreifuss muscular dystrophy. AB - Emerin and specific isoforms of nesprin-1 and -2 are nuclear membrane proteins which are binding partners in multi-protein complexes spanning the nuclear envelope. We report here the characterisation of the residues both in emerin and in nesprin-1alpha and -2beta which are involved in their interaction and show that emerin requires nesprin-1 or -2 to retain it at the nuclear membrane. Using several protein-protein interaction methods, we show that residues 368 to 627 of nesprin-1alpha and residues 126 to 219 of nesprin-2beta, which show high homology to one another, both mediate binding to emerin residues 140-176. This region has previously been implicated in binding to F-actin, beta-catenin and lamin A/C suggesting that it is critical for emerin function. Confirmation that these protein domains interact in vivo was shown using GFP-dominant negative assays. Exogenous expression of either of these nesprin fragments in mouse myoblast C2C12 cells displaced endogenous emerin from the nuclear envelope and reduced the targeting of newly synthesised emerin. Furthermore, we are the first to report that emerin mutations which give rise to X-linked Emery-Dreifuss muscular dystrophy, disrupt binding to both nesprin-1alpha and -2beta isoforms, further indicating a role of nesprins in the pathology of Emery-Dreifuss muscular dystrophy. PMID- 17462628 TI - Membrane expression of DR4, DR5 and caspase-8 levels, but not Mcl-1, determine sensitivity of human myeloma cells to Apo2L/TRAIL. AB - The improved recombinant form of the death ligand Apo2L/TRAIL (Apo2L/TRAIL.0) is not cytotoxic for normal human cells and is a good candidate for the therapy of multiple myeloma (MM), a B-cell neoplasia that remains incurable. We have analyzed the molecular determinants of myeloma sensitivity to Apo2L/TRAIL.0 in a number of MM cell lines, the mechanisms of resistance and a possible way of overcoming it. Expression of one death receptor for Apo2L/TRAIL (DR4 or DR5) is sufficient to transduce death signals, though DR5 was more efficient when both receptors were present. Membrane expression of decoy receptors (DcR1, DcR2) and intracellular levels of c-FLIP(L), XIAP and Mcl-1 were not predictive of resistance to Apo2L/TRAIL. Inhibition of Mcl-1 degradation did not prevent Apo2L/TRAIL-induced apoptosis. In IM-9 cells, resistance was associated to a reduced caspase-8 expression. U266 cells, though expressing significant levels of DR4 and caspase-8, were nevertheless resistant to Apo2L/TRAIL. This resistance could be overcome by co-treatment with valproic acid (VPA), a histone deacetylase inhibitor. VPA caused the redistribution of DR4 to plasma membrane lipid rafts and restored DR4 signaling. Overexpression of Mcl-1 in U266 cells did not prevent Apo2L/TRAIL cytotoxicity in VPA-sensitized cells. These results, taken together, support the possible use of Apo2L/TRAIL.0 in the treatment of MM. PMID- 17462629 TI - Involvement of BTBD1 in mesenchymal differentiation. AB - BTBD1 is a recently cloned BTB-domain-containing protein particularly expressed in skeletal muscle and interacting with DNA topoisomerase 1 (Topo1), a key enzyme of cell survival. We have previously demonstrated that stable overexpression of a N-terminal truncated BTBD1 inhibited ex vivo myogenesis but not adipogenesis of pluripotent C2C12 cells. Here, BTBD1 expression was studied in three models of cellular differentiation: myogenesis (C2C12 cells), adipogenesis (3T3-L1 cells) and osteogenesis (hMADS cells). BTBD1 mRNA was found to be upregulated during myogenesis. At the opposite, we have not observed BTBD1 upregulation in an altered myogenesis cellular model and we observed a downregulation of BTBD1 mRNA expression in adipogenesis. Interestingly, amounts of Topo1 protein, but not Topo1 mRNA, were found to be modulated at the opposite of BTBD1 mRNA. No variation of BTBD1 expression was measured during osteogenesis. Taken together, these results indicate that BTBD1 mRNA is specifically regulated during myogenic and adipogenic differentiation, in relation with Topo1 expression. Moreover, they corroborate observations made previously with truncated BTBD1 and show that BTBD1 is a key protein of balance between adipogenesis and myogenesis. Finally, a transcriptome analysis gave molecular clues to decipher BTBD1 role, with an emphasis on the involvement in ubiquitin/proteasome degradation pathway. PMID- 17462630 TI - Central neural distribution of immunoreactive Fos and CRH in relation to plasma ACTH and corticosterone during sepsis in the rat. AB - Although brain pathways activated by sepsis may respond acutely to endotoxin administration, the long-term central response to sepsis is not known. We prepared male rats for hormonal sampling at the circadian nadir (AM) and peak (PM) after cecal ligation and puncture (CLP) or sham surgery. Diurnal variation of corticosterone was present on postoperative day (D) 3 and D4 after sham surgery but not after CLP. CLP increased Fos immunostaining in the nucleus of tractus solitarius (NTS), ventrolateral medulla, medullary raphe, parabrachial nucleus, hypothalamus, amygdala, bed nucleus of stria terminalis, and preoptic region. Fos responses were generally greatest on D1 but persisted to the AM of D4. The number of Fos-positive cell nuclei in the NTS on D3 and D4 did not differ but had greater variance on D3 than on D4 (P<0.01) with a divergent response in the PM of D3 that was correlated with plasma ACTH (r=0.927, P<0.01) but not with corticosterone. CLP increased CRH-staining intensity in the hypothalamic paraventricular neurons uniformly from D1 through D4 (P<0.01). Similar to Fos in NTS, this response was correlated with plasma ACTH (r=0.738, P<0.05) and adrenal size (r=0.730, P<0.05) in the PM of D3. Neuronal CRH became detectable after CLP in specific medullary areas on D1 and in the preoptic region on D3 and D4. Thus, the suppression of circadian variation by CLP was associated with central neural responses that increased in relation to plasma ACTH without apparent influence on the release of corticosterone. PMID- 17462631 TI - Echinostoma caproni: identification of enolase in excretory/secretory products, molecular cloning, and functional expression. AB - In order to investigate molecules that could be involved in host-trematode relationships, we have analysed the excretory/secretory products (ESP) of Echinostoma caproni following a proteomic approach. Actin, Gluthathione S transferase (GST) and enolase have been identified in the ESP. Enolase, observed to be one of the most abundant proteins, was further characterized. The molecular cloning and in vitro expression in Escherichia coli of E. caproni enolase allowed us to determine that the protein contains 431 amino acids and a theoretical MW of 46272 Da. E. caproni enolase shows high homology to other trematode enolases. The recombinant protein binds specifically to human plasminogen in vitro, as observed for the native protein, confirming its properties as a host-interacting molecule. PMID- 17462632 TI - Phytomonas: transport of amino acids, hexoses and polyamines. AB - Phytomonas cells (Phytomonas Jma) isolated from the latex of Jatropha macrantha were assayed for amino acid, hexose and polyamine transport. Results showed high transport rates for glucose and fructose (193 and 128 pmol min(-1) 10(-7) cells, respectively) and lower, but significant rates, for proline, arginine, cysteine and glutamate (between 1.7 and 5.8 pmol min(-1) 10(-7) cells). Minor transport activities were observed for serine, glycine and aspartate (<1 pmol min(-1) 10( 7) cells). Amino acid transport processes do not seem to be regulated by starvation or during the growth phases. Polyamine transport was also evaluated showing a clear preference for spermidine over putrescine (3.4 and 0.4 pmol min( 1) 10(-7) cells, respectively). This work represents the first report on metabolite transport in phytomonads. PMID- 17462633 TI - Somatostatin induces rapid contraction of neuroendocrine cells. AB - The peptide hormone somatostatin, as well as the somatostatin analog octreotide, induces rapid morphological changes in neuroendocrine cells. The effect can be detected in less than 2 min: retraction fibers are formed, cells round up and cell-cell contacts are broken. Somatostatin-dependent cell contraction is inhibited by Y-27632, indicating that this effect is dependent on Rho kinase. In BON1 cells, the somatostatin-induced inhibition of forskolin-induced secretion of chromogranin A is not blocked by Y-27632. It is therefore concluded that the inhibitory effect of somatostatin in forskolin-stimulated cells is not dependent on cell contraction. PMID- 17462634 TI - Extensive selection for the enrichment of G4 DNA motifs in transcriptional regulatory regions of warm blooded animals. AB - A comprehensive analysis of potential G4 DNA motifs (G4Ms) in genomic regions flanking transcription start sites (TSS) was performed across 13 animal species. We found that G4Ms are significantly enriched in the transcriptional regulatory regions (TRRs) of warm-blooded animals. Further analysis of human genes in different temporal groups reveals that the enrichment is not specific to genes found only in warm-blooded species but instead exist in a wide range of genes. Our findings therefore suggest that the high prevalence of G4Ms in TRRs is extensively selected in warm-blooded animals, supporting the hypothesis that G4Ms are involved in the regulation of gene transcription. PMID- 17462635 TI - Transition metal transport. AB - Transition metal transporters are of central importance in the plant metal homeostasis network which maintains internal metal concentrations within physiological limits. An overview is given of the functions of known transition metal transporters in the context of the unique chemical properties of their substrates. The modifications of the metal homeostasis network associated with the adaptation to an extreme metalliferous environment are illustrated in two Brassicaceae metal hyperaccumulator model plants based on cross-species transcriptomics studies. In a comparison between higher plants and unicellular algae, hypotheses are generated for evolutionary changes in metal transporter complements associated with the transition to multicellularity. PMID- 17462636 TI - Roles of ion channels and transporters in guard cell signal transduction. AB - Stomatal complexes consist of pairs of guard cells and the pore they enclose. Reversible changes in guard cell volume alter the aperture of the pore and provide the major regulatory mechanism for control of gas exchange between the plant and the environment. Stomatal movement is facilitated by the activity of ion channels and ion transporters found in the plasma membrane and vacuolar membrane of guard cells. Progress in recent years has elucidated the molecular identities of many guard cell transport proteins, and described their modulation by various cellular signal transduction components during stomatal opening and closure prompted by environmental and endogenous stimuli. PMID- 17462637 TI - Serum anti-Mullerian hormone and inhibin B in disorders of spermatogenesis. AB - This study investigates circulating concentrations of AMH and inhibin B in men with azoospermia. Serum AMH and inhibin B are significantly lower in the men with nonobstructive azoospermia compared to the controls and the men with obstructive azoospermia, suggesting that these hormones could be markers of nonobstructive azoospermia. PMID- 17462638 TI - Limiting access to letrozole--is it justified? AB - Letrozole is an effective drug for ovulation induction and superovulation. Limiting its access is a disservice to our infertile patients and to the advancement in ovulation-related research. PMID- 17462639 TI - The use of gonadotropin-releasing hormone (GnRH) agonist to induce oocyte maturation after cotreatment with GnRH antagonist in high-risk patients undergoing in vitro fertilization prevents the risk of ovarian hyperstimulation syndrome: a prospective randomized controlled study. AB - OBJECTIVE: To determine whether there are any differences in the incidence of ovarian hyperstimulation syndrome (OHSS) and implantation rates in high-risk patients undergoing IVF using a protocol consisting of GnRH agonist trigger after cotreatment with GnRH antagonist or hCG trigger after dual pituitary suppression protocol. DESIGN: Prospective randomized controlled trial. SETTING: University based tertiary fertility center. PATIENT(S): Sixty-six patients under 40 years of age with polycystic ovarian syndrome, polycystic ovarian morphology, or previous high response undergoing IVF. INTERVENTION(S): Patients were randomized to an ovarian stimulation protocol consisting of either GnRH agonist trigger after cotreatment with GnRH antagonist (study group) or hCG trigger after dual pituitary suppression with a GnRH agonist (control group). Both groups received luteal phase and early pregnancy supplementation with IM progesterone (P), and patients in the study group also received E(2) patches and their doses were adjusted according to the serum levels. MAIN OUTCOME MEASURE(S): Incidence of OHSS and implantation rate. RESULT(S): None of the patients in the study group developed any form of OHSS compared with 31% (10/32) of the patients in the control group. There were no significant differences in the implantation (22/61 [36.0%] vs. 20/64 [31.0%]), clinical pregnancy (17/30 [56.7%] vs. 15/29 [51.7%]), and ongoing pregnancy rates (16/30 [53.3%] vs. 14/29 [48.3%]) between the study and control groups, respectively. CONCLUSION(S): The use of a protocol consisting of GnRH agonist trigger after GnRH antagonist cotreatment combined with adequate luteal phase and early pregnancy E(2) and P supplementation reduces the risk of OHSS in high-risk patients undergoing IVF without affecting implantation rate. PMID- 17462640 TI - Whole genome deoxyribonucleic acid microarray analysis of gene expression in ectopic versus eutopic endometrium. AB - OBJECTIVE: To use DNA microarrays to identify differentially expressed genes in eutopic endometrium compared with ectopic endometrium. DESIGN: Prospective, cross sectional, observational study. SETTING: University Medical Center and Research Laboratory. PATIENT(S): Eleven women with endometriosis. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Differential gene expression. RESULT(S): Seven hundred seventeen of the 53,000 probes on the whole human DNA microarrays were changed by twofold or greater in ectopic versus eutopic endometrium. Families of genes that were expressed differentially include genes that code for proteins associated with the immune system and inflammatory pathways, cell adhesion, cell-cell junctions, the extracellular matrix and its remodeling, cytoskeletal proteins, and signal transduction pathway components, among others. CONCLUSION(S): The altered immune environment may allow survival of endometriotic cells that enter the abdominal cavity. Alterations of cell adhesion-associated genes may contribute to the adhesive and invasive properties of ectopic endometrium, and changes in signal transduction pathways support a change in the communication among cells of the endometrial explant compared with eutopic endometrium. These families of differentially expressed genes provide multiple opportunities for the development and testing of new hypotheses regarding endometriosis. PMID- 17462641 TI - Risk of metabolic complications in the new PCOS phenotypes based on the Rotterdam criteria. AB - OBJECTIVE: To determine the risk of metabolic complications, primarily metabolic syndrome, in all polycystic ovary syndrome (PCOS) phenotypes compared with control subjects. DESIGN: Retrospective chart review. SETTING: University practice. PATIENT(S): Women with PCOS (Rotterdam definition; n = 258) and women without PCOS seen during the same time period for an annual exam used as controls (n = 110). INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Metabolic syndrome. RESULT(S): Three PCOS phenotypes had a significantly higher prevalence of metabolic syndrome compared with the control subjects: oligomenorrhea/oligo ovulation (O) + hyperandrogenism (H) + polycystic ovaries (P), age-adjusted odds ratio [OR] 6.3 (95% confidence interval 2.1-18.9); O+H, OR 7.8 (2.2-27.5); and H+P, OR 8.2 (2.3-29.3). There was no significant difference in the prevalence of metabolic syndrome between women with O+P and control subjects, even in obese women. The prevalence of insulin resistance and glucose intolerance was not significantly different between POCS phenotypes CONCLUSION(S): The risk of metabolic syndrome may vary among the four phenotypes of PCOS based on the Rotterdam criteria. This new information may be of relevance in counseling women with PCOS although larger studies may be needed to validate our findings. PMID- 17462642 TI - Intelligent, impedance-regulated, pulsed coagulation in a porcine renal artery model. AB - OBJECTIVE: To compare the efficacy of conventional pulsed coagulation (CPC) and newly developed intelligent, impedance-regulated, pulsed coagulation (IPC) in the sealing of porcine renal arteries. DESIGN: Prospective, randomized experimental study. SETTING: Isolated porcine artery model in an academic research environment. ANIMAL(S): Female Swabian Hall pigs. INTERVENTION(S): Renal arteries were harvested from Swabian pigs, flushed with saline, and sealed with bipolar open forceps by using high-frequency modulations of CPC (CPC-I: 800-ms pulse, 30 ms pause; CPC-II: 800-ms pulse, 300-ms pause) or IPC (self-regulation of the current flow to tissue impedance during thermal alteration). Additional vessels underwent multiple CPC. Burst pressure and seal failure were measured by increasing the pressure in the sealed arteries with saline infusion until rupture of the seal or the vessel wall. MAIN OUTCOME MEASURE(S): Mean burst pressure, number of instant and secondary seal failures, and relation of burst pressure to vessel diameter. RESULT(S): Mean burst pressure after IPC (585.5 +/- 56.8 mm Hg) was statistically significantly higher than that after CPC (CPC-I: 372.6 +/- 40.0 mm Hg; CPC-II: 334.2 +/- 44.2 mm Hg). Only 5.0% of the vessel seals after IPC, but 34.0% and 39.5% after CPC-I and CPC-II, showed instant or secondary seal failures, which also was a statistically significant difference. Seal quality after multiple CPC was comparable to that observed after the single IPC application (burst pressure, 597.3 +/- 60.1 [MCPC-I] mm Hg and 656.2 +/- 56.5 mm Hg [MCPC-II]; seal failure rate, 0). CONCLUSION(S): In an isolated porcine renal artery model, self-regulating modulation of energy-based vessel coagulation achieved superior thermal fusion of vascular tissue than did CPC. This promising novel technique should be analyzed further to determine its in vivo efficacy in long-term studies. PMID- 17462643 TI - Serum 17-hydroxyprogesterone strongly correlates with intratesticular testosterone in gonadotropin-suppressed normal men receiving various dosages of human chorionic gonadotropin. AB - OBJECTIVE: To determine if serum concentrations of testosterone precursors would correlate with intratesticular testosterone (ITT) concentration measured directly by testicular aspiration and allow for a less invasive means of inferring ITT. DESIGN: Controlled clinical study. SETTING: Healthy volunteers in an academic research environment. PATIENT(S): Twenty-nine normal men. INTERVENTION(S): We determined ITT concentration by testicular aspiration before and after treatment in men receiving exogenous T to block endogenous gonadotropin production and randomly assigned to one of four doses of hCG (0, 125 IU, 250 IU, or 500 IU every other day) for 3 weeks. MAIN OUTCOME MEASURE(S): The association between serum 17 hydroxyprogesterone (17OH-P), androstenedione, and DHEA and ITT. RESULT(S): With T administration alone, serum 17OH-P decreased significantly and increased significantly when 500 IU hCG was administered. End-of-treatment ITT strongly correlated with serum 17OH-P. Moreover, serum 17OH-P, but not androstenedione or DHEA, was independently associated with end-of-treatment ITT by multivariate linear regression. CONCLUSION(S): Serum 17OH-P is highly correlated with ITT in gonadotropin-suppressed normal men receiving T and stimulated with hCG. Serum 17OH-P is a surrogate biomarker of ITT and may be useful in research and in men receiving gonadotropin therapy for infertility. PMID- 17462644 TI - The influence of social status on the rate of growth, eye color pattern and insulin-like growth factor-I gene expression in Nile tilapia, Oreochromis niloticus. AB - Many aspects of teleost physiology are subject to regulation by social interactions. To evaluate the relationship of social status with growth, eye color pattern and hepatic Insulin-like Growth Factor-I (IGF-I) mRNA expression, 30 Oreochromis niloticus were isolated for 10 days and were used in a social pair study. Results revealed that growth of both dominant (except 1 day after social interaction) and subordinate individuals was suppressed, but growth suppression was greater in the subordinates. The dominant fish completely inhibited the feeding of the subordinate individuals during and 1 day after they were introduced into the aquaria together. After that, a pattern of highly aggressive attacks by dominant fish only partially inhibited feeding by the subordinates. Differential alterations in growth rate between dominants and subordinates were attributed more to behavioral changes (i.e., feeding) as transduced by physiological regulators (i.e., IGF-I level and possibly serotonin and/or neuropeptide Y) but may also be due to changes in metabolism. The fish's relative position in the social hierarchy consistently influenced the levels of IGF-I mRNA in the liver and the eye color pattern. Lower social status depressed hepatic IGF I levels while dominant status stimulated hepatic IGF-I production, possibly in response to inhibition of somatostatin release in the hypothalamus, leading to greater secretion of pituitary growth hormone (GH). A significant positive association was detected between the IGF-I mRNA expression of the dominant fish and the level of aggression (number of attacks) during the encounter. Social status also influenced the eye color pattern of the fish. During aggressive interactions, most of the fish (22 out of 24) displayed decreased eye darkening. At the later part of the encounter, all subsequent subordinates displayed eye darkening patterns which acted as a social signal announcing social submission. After the encounter dominant fish had paler eye color pattern than subordinates. PMID- 17462645 TI - Elevated rates of testosterone-related disorders in women with autism spectrum conditions. AB - The androgen theory of autism proposes that autism spectrum conditions (ASC) are in part due to elevated fetal testosterone (FT) levels, which are positively correlated with a number of autistic traits and inversely correlated with social development and empathy. A medical questionnaire was completed by n=54 women with ASC, n=74 mothers of children with ASC, and n=183 mothers of typically developing children to test whether women with ASC have an increased rate of testosterone related medical conditions, and to see whether mothers of children with ASC show similar abnormalities, as part of the 'broader autism phenotype'. Compared to controls, significantly more women with ASC reported (a) hirsutism, (b) bisexuality or asexuality, (c) irregular menstrual cycle, (d) dysmenorrhea, (e) polycystic ovary syndrome, (f) severe acne, (g) epilepsy, (h) tomboyism, and (i) family history of ovarian, uterine, and prostate cancers, tumors, or growths. Compared to controls, significantly more mothers of ASC children reported (a) severe acne, (b) breast and uterine cancers, tumors, or growths, and (c) family history of ovarian and uterine cancers, tumors, or growths. These results suggest current hormone abnormalities in women with ASC and their mothers. Direct investigations of serum testosterone levels and genetic susceptibility to high testosterone production or sensitivity in women with ASC would illuminate the origin of these conditions. The relationship between FT and current testosterone levels also needs to be clarified. The present results may be relevant to understanding the increased male risk to developing autism. PMID- 17462646 TI - Evidence for the influence of testosterone in the performance of spatial navigation in a virtual water maze in women but not in men. AB - Testosterone (T) may be associated with enhanced spatial navigation in a number of rodent species, although the nature of the relation is equivocal. Similarly, numerous studies in humans generally have found that T is associated with enhanced spatial ability on a variety of paper and pencil tasks that may relate to navigational ability. However, relatively few studies have reported effects of T on navigational ability in humans. We investigated the relationship between endogenous T and performance on a virtual water maze (vWM) and mental rotations test (MROT). ELISA for T was performed on salivary samples that were obtained from participants before and after completion of both spatial tasks. Results indicated that women with low T required more time to locate the hidden platform in the vWM than either group of men or women with high T. Significant negative correlations were found for the entire sample between vWM performance and T, and between vWM latency to escape and MROT. Similar significant correlations were found in women but not men. Thus, our data support the position that T improves performance in the vWM in a linear fashion, most strongly in women. However, further work is needed to confirm this hypothesis in humans. PMID- 17462647 TI - Individual differences in novelty-seeking and emotional reactivity correlate with variation in maternal behavior. AB - Numerous studies have demonstrated that Sprague-Dawley rats exhibit a wide range of locomotor reactivity when placed in a novel environment. High Responder (HR) rats show exaggerated locomotor response to novelty, enhanced neuroendocrine stress reactivity, decreased anxiety-like behavior, and propensity to self administer psychostimulants, compared to the less active Low Responder (LR) animals. Few studies have explored the early environmental factors which may underlie the HR-LR differences in emotional reactivity. Considering the enormous impact of maternal care on rodent neurodevelopment, we sought to examine maternal behavior in HR-LR dams to determine whether they exhibit differences which could contribute to their offspring's differential temperaments. Females, like males, can be classified as HR versus LR, showing marked differences in novelty-induced locomotor activity and anxiety-like behavior. HR-LR mothers behaved differently with their pups during the first two postpartum weeks. LR dams spent greater time licking and nursing their pups compared to HR dams, with the most prominent differences occurring during the second postpartum week. By contrast, when non lactating HR-LR females were presented with orphaned pups, the pattern of maternal response was reversed. HR females were more responsive and showed greater maternal care of the novel pups compared to LR females, which were probably inhibited due to fear of the unfamiliar pups. This underscores the critical interplay between the female's emotional phenotype, her hormonal status and her familiarity with the pup as key factors in determining maternal behavior. Future work should explore neural and hormonal mechanisms which drive these HR-LR differences in maternal behavior and their impact on the development of the offspring. PMID- 17462648 TI - Celiac disease in intrauterine growth restriction. PMID- 17462649 TI - Attitudes of doctors toward establishing safe abortion units in Ghana. AB - OBJECTIVES: To assess physician knowledge of the current legal status of abortion in Ghana and determine the proportion physicians in favor of establishing units where safe abortion would be provided, and the proportion of those willing to offer such services. METHODS: A survey conducted at Komfo Anokye Teaching Hospital, Ghana, in August 2003. Using a self-administered questionnaire, 74 randomly selected physicians expressed their views on whether abortion units should be established within national health facilities in Ghana, and what role they would play in them. RESULTS: Of the 59 physicians (80%) who favored the establishment of safe abortion units within national health facilities, 27 (36%) indicated a willingness to take part in counseling only, 33 (45%) were prepared to carry out abortions, and 14 (19%) said they would play no role in these units. CONCLUSION: Most physicians participating in this study conducted in Ghana were willing to play some role in the provision of safe abortion services. PMID- 17462650 TI - HPV-related vulvar intraepithelial neoplasia: outcome of different management modalities. AB - OBJECTIVE: To evaluate the outcome of various management schemes for HPV-related vulvar intraepithelial neoplasia (VIN, usual type). METHODS: Retrospective chart review of patients with histologically diagnosed grade 2/3-VIN who had at least one year of follow-up. The variables that were collected included patient characteristics, management modalities, and clinical outcome. RESULTS: Fifty patients with a median age of 45 years old were evaluated. The median duration of follow-up was 43.5 months (12-186). Complete response (CR) and partial response occurred in 28 (56%) and 4 (8%), respectively. Nineteen of 28 patients with CR recurred with VIN. Surgical excision yielded higher CR (77%) than did either ablational techniques (21-33%) or topical immunotherapy (33%). CONCLUSION: In this experience, surgical excision for VIN, usual type, resulted in better therapeutic success rates than other treatment modalities. Management schemes should be individualized based on extent of disease and patient compliance. PMID- 17462651 TI - In vitro miconazole susceptibility and clinical outcome in vulvovaginal candidiasis. PMID- 17462652 TI - The echogenecity of the intima-media complex in the common carotid artery is closely related to the echogenecity in plaques. AB - OBJECTIVE: The echogenecity measured by ultrasound of atherosclerotic plaques is related to future cardiovascular events. The aim of the present study is to relate the grey scale median of the intima-media complex (IM-GSM) of the common carotid artery (CCA) to the echogenecity of carotid plaques. MATERIAL AND RESULTS: In the Prospective Study of the Vasculature in Uppsala Seniors (PIVUS) study, a population-based study of 1016 subjects aged 70, carotid artery intima media thickness (IMT) and IM-GSM were evaluated by ultrasound and computerized analysis. Also the occurrence of plaque and plaque GSM were measured. The echogenecity of the plaques was also visually estimated by the Gray-Weale classification. In subjects with a carotid plaque (n=582), IM-GSM in CCA was correlated to GSM in the plaque (r=0.60, p<0.0001) independently of plaque size and IMT. IM-GSM in CCA was also correlated to the visually estimated echogenecity (p<0.0001 for trend). CONCLUSION: IM-GSM of the CCA is closely related to the echogenecity in overt carotid plaques, regardless if evaluated by the same computerized method or evaluated visually. This finding suggests that IM-GSM of CCA could be an important and easily measurable characteristic of the carotid artery wall that could be obtained in almost all subjects and not only those with an overt plaque. PMID- 17462653 TI - Genetic contribution to brachial artery flow-mediated dilation: the Northern Manhattan Family Study. AB - BACKGROUND: Brachial artery flow-mediated dilation (FMD) is a non-invasive measure of endothelial function. Endothelial dysfunction has been associated with traditional vascular risk factors and increased risk of cardiovascular disease. The importance of genetic contribution to FMD and baseline brachial artery diameter has not been shown in Hispanic populations. The purpose of this study was to estimate the heritability of FMD. METHODS: Flow mediated dilation and brachial artery diameter were measured in a subset of Caribbean Hispanic families from the ongoing Northern Manhattan Family Study (NOMAFS), which studies the contribution of genetics to stroke and cardiovascular risk factors. The age- and sex-adjusted heritability of FMD was estimated using variance component methods. RESULTS: The current data include 620 subjects (97 probands and 523 relatives) from 97 families. The age and sex-adjusted heritability of brachial artery diameter was 0.57 (p<0.01). The age- and sex-adjusted heritability of FMD was 0.20 (p=0.01). After additional adjustment for systolic and diastolic blood pressure, body mass index, smoking, lipid, diabetes mellitus, medication, and baseline brachial artery diameter, the heritability of FMD was 0.17 (p=0.01). CONCLUSIONS: We found modest heritability of FMD. FMD might be a reasonable phenotype for further investigation of genetic contribution to atherosclerosis. PMID- 17462654 TI - Remarkable increase of apolipoprotein B48 level in diabetic patients with end stage renal disease. AB - Apolipoprotein (apo) B48 is a structural protein of chylomicrons. Fasting serum levels of apoB48 suggest the presence of small number of remnant chylomicron particles which are thought to be an atherogenic lipoprotein. In view of the high incidence of coronary heart disease (CHD) in patients with diabetic nephropathy, we decided to measure the plasma apoB48 level in type 2 diabetics with diabetic nephropathy at various stages to ascertain how apoB48 relates to the progression of diabetic nephropathy. Patients with type 2 diabetes (n=105) were stratified into four groups: normo-albuminuria, micro-albuminuria, overt-proteinuria, and patients with end-stage renal disease (ESRD) receiving hemodialysis. Age-matched diabetic hypertensive patients (n=24) and non-diabetic ESRD patients on hemodialysis (n=47) were also enrolled. Plasma triglyceride (TG) levels rose as diabetic nephropathy progressed to overt-proteinuria. No further elevation in TG was observed in diabetic ESRD, however, and the TG levels were normal in non diabetic ESRD. A similar pattern was observed for remnant-like particle cholesterol (RLP-C). In contrast to the changes observed for TG and RLP-C, the levels of apoB48 increased steadily as the diabetic nephropathy progressed (control, 3.7; normo, 5.7; micro, 6.9; overt, 10.6 mg/l, respectively). ApoB48 peaked in the diabetic ESRD (19 mg/l) and was also markedly elevated in non diabetic ESRD (10.1mg/l). The apoB48/TG and apoB48/total-apoB ratios were substantially elevated in both diabetic and non-diabetic ESRD. These results are the first to demonstrate remarkable elevations of plasma apoB48 in patients with both diabetic and non-diabetic ESRD. The remarkably high level of apoB48 in diabetic ESRD seems to be attributable to dyslipidemia induced by both diabetic nephropathy and ESRD. PMID- 17462655 TI - Soft-tissue artefact assessment during step-up using fluoroscopy and skin-mounted markers. AB - When measuring knee kinematics with skin-mounted markers, soft tissue and structures surrounding the knee hide the actual underlying segment kinematics. Soft-tissue artefacts can be reduced when plate-mounted markers or marker trees are used instead of individual unconstrained mounted markers. The purpose of this study was to accurately quantify the soft-tissue artefacts and to compare two marker cluster fixation methods by using fluoroscopy of knee motion after total knee arthroplasty during a step-up task. Ten subjects participated 6 months after their total knee arthroplasty. The patients were randomised into (1) a plate mounted marker group and (2) a strap-mounted marker group. Fluoroscopic data were collected during a step-up motion. A three-dimensional model fitting technique was used to reconstruct the in vivo 3-D positions of the markers and the implants representing the bones. The measurement errors associated with the thigh were generally larger (maximum translational error: 17mm; maximum rotational error 12 degrees ) than the measurement errors for the lower leg (maximum translational error: 11mm; maximum rotational error 10 degrees ). The strap-mounted group showed significant more translational errors than the plate-mounted group for both the shank (respectively, 3+/-2.2 and 0+/-2.0mm, p = 0.025) and the thigh (2+/-2.0 and 0+/-5.9mm, p = 0.031). The qualitative conclusions based on interpretation of the calculated estimates of effects within the longitudinal mixed-effects modelling evaluation of the data for the two groups (separately) were effectively identical. The soft-tissue artefacts across knee flexion angle could not be distinguished from zero for both groups. For all cases, recorded soft-tissue artefacts were less variable within subjects than between subjects. The large soft-tissue artefacts, when using clustered skin markers, irrespective of the fixation method, question the usefulness of parameters found with external movement registration and clinical interpretation of stair data in small patient groups. PMID- 17462656 TI - MRI-derived body segment parameters of children differ from age-based estimates derived using photogrammetry. AB - Body segment parameters are required when researching joint kinetics using inverse dynamics models. However, the only regression equations for estimating pediatric body segment parameters across a wide age range were developed, using photogrammetry, based on 12 boys and have not been validated to date (Jensen, R.K., 1986. Body segment mass, radius and radius of gyration proportions of children. Journal of Biomechanics 19, 359-368). To assess whether these equations could validly be applied to girls, we asked whether body segment parameters estimated by the equations differ from parameters measured using a validated magnetic resonance imaging (MRI) method. If so, do the differences cause significant differences in joint kinetics during normal gait? Body segment parameters were estimated from axial MRIs of the left thigh and shank of 10 healthy girls (9.6 +/- 0.9 years) and compared to those from Jensen's equations. Kinematics and kinetics were collected for 10 walking trials. Extrema in hip and knee moments and powers were compared between the two sets of body segment parameters. With the exception of the shank mass center and radius of gyration, body segment parameters measured using MRI were significantly different from those estimated using regression equations. These systematic differences in body segment parameters resulted in significant differences in sagittal-plane joint moments and powers during gait. Nevertheless, it is doubtful that even the greatest differences in kinetics are practically meaningful (0.3% BW x HT and 0.7% BW x HT/s for moments and power at the hip, respectively). Therefore, body segment parameters estimated using Jensen's regression equations are a suitable substitute for more detailed anatomical imaging of 8-10-year-old girls when quantifying joint kinetics during gait. PMID- 17462657 TI - An alternative approach to normalization and evaluation for gait patterns: Procrustes analysis applied to the cyclograms of sprinters and middle-distance runners. AB - In order to compare gait patterns, a common procedure is to normalize strides both in time and magnitude. The stride duration is usually normalized to a time percentage before averaging curves. As the timing of event occurrences may shift across strides, the shape of the averaged curves is distorted and therefore the standard deviation is overvalued. Stride magnitude normalization is performed by means of dimensionless numbers. However, there is little agreement on which body size correction methods should be used. The Procrustes method describes curve shape and shape change in a mathematical and statistical framework, independently of time and size factors. The present study aims to explore how this technique may be used for time- and magnitude-stride normalization to reflect individual and group mean responses. The Procrustes method, which combines quantitative and visual features, is applied to the shape of the ankle and knee cyclograms. Superimposition of 25 cyclograms (10 for sprinters (SP) and 15 for middle distance runners (MDR)) was supplemented by statistical procedures (principal component analysis, discriminant function) to extract the main key events, which vary according to the athletic specialities. In comparison with the MDR (poulaine shaped cyclogram), the ovoid cyclogram of SP reveals the following gait indicators: a short braking phase, a rapid initial lower limb swing in the forward direction, a fast upward movement of the knee and ankle, and an active foot contact. The Procrustes approach could be used to describe other quasi periodic movements through relative motion plots (e.g., cyclograms, angle-angle diagrams, phase plane portraits). PMID- 17462658 TI - Antibody neutralization of TGF-beta enhances the deterioration of collagen fascicles in a tissue-cultured tendon matrix with ex vivo fibroblast infiltration. AB - A tissue-cultured tendon matrix infiltrated with cultured fibroblasts can be regarded as an ideal tissue-engineered tendon model. To clarify the role of TGF beta in a tissue-cultured tendon matrix during ex vivo cellular infiltration, the present ex vivo study was conducted to test the following hypothesis that antibody neutralization of TGF-beta enhances weakening of the collagen fascicles of the patellar tendon matrix in response to ex vivo fibroblast infiltration. In skeletally mature female rabbits, fibroblasts were isolated from the right patellar tendons using an explant culture technique, and the left patellar tendons underwent multiple freeze/thaw treatment with liquid nitrogen to obtain an acellular tendon matrix. Each acellular tendon was placed in a collagen gel containing cultured fibroblasts and then incubated with or without anti-TGF-beta1 antibody for 6 weeks. We found that antibody neutralization of TGF-beta enhanced the decrease in the tensile strength and tensile modulus of the collagen fascicles of the patellar tendon matrix in response to ex vivo fibroblast infiltration. The present study indicates a possibility that TGF-beta may have a role in suppressing the material deterioration of the fascicles in the tendon during ex vivo cellular infiltration. PMID- 17462659 TI - In vivo knee moments and shear after total knee arthroplasty. AB - Tibiofemoral loading is very important in cartilage degeneration as well as in component survivorship after total knee arthroplasty. We have previously reported the axial knee forces in vivo. In this study, a second-generation force-sensing device that measured all six components of tibial forces was implanted in a 74 kg, 83-year-old male. Video motion analysis, ground reaction forces, and knee forces were measured during walking, stair climbing, chair-rise, and squat activities. Peak total force was 2.3 times body weight (BW) during walking, 2.5 x BW during chair rise, 3.0 x BW during stair climbing, and 2.1 x BW during squatting. Peak anterior shear force at the tibial tray was 0.30 x BW during walking, 0.17 x BW during chair rise, 0.26 x BW during stair climbing, and 0.15 x BW during squatting. Peak flexion moment at the tray was 1.9% BW x Ht (percentage of body weight multiplied by height) for chair-rise activity and 1.7% BW x Ht for squat activity. Peak adduction moment at the tray was -1.1% BW x Ht during chair rise, -1.3% BW x Ht during squatting. External knee flexion and adduction moments were substantially greater than flexion and adduction moments at the tray. The axial component of forces predominated especially during the stance phase of walking. Shear forces and moments at the tray were very modest compared to total knee forces. These findings indicate that the soft tissues around the knee absorbed most of the external shear forces. Our results highlight the importance of direct measurements of knee forces. PMID- 17462660 TI - Preparation of stationary phases for reversed-phase high-performance liquid chromatography using thermal treatments at high temperature. AB - Batches of poly(methyloctylsiloxane) (PMOS)-loaded silica were prepared by deposition from a solution of PMOS into the pores of HPLC silica. Portions of PMOS-loaded silica were subjected to a thermal treatment at 100 degrees C for 24h (condition 1) in a tube furnace under a nitrogen atmosphere. After that, the material was heated for 4h at higher temperatures (150-400 degrees C) (condition 2). Heating at higher temperatures produces polymer bilayers. Non-immobilized and thermally treated stationary phases were characterized by percent carbon, (29)Si cross-polarization magic angle spinning nuclear magnetic resonance spectroscopy and reversed-phase chromatographic performance. The results show that thermal treatment between 150 and 300 degrees C accelerates the immobilization process, possibly due to some bond breaking of the polysiloxane, with formation of strong linkages to the surface of the support, resulting in more complete coverage of the silica. The chromatographic results show an improvement of efficiency with the increase of the temperature of condition 2 up to 300 degrees C and an increase in the resolution of the components, mainly for the phase heated at 300 degrees C. Such results demonstrate that a two-step thermal treatment (100 degrees C then 150-300 degrees C) produces stationary phases with good properties for use in reversed-phase high-performance liquid chromatography. PMID- 17462661 TI - Use of headspace solid-phase microextraction for the quantification of poly(3 hydroxybutyrate) in microbial cells. AB - Biopolymers such as poly(hydroxyalkanoates) (PHAs) have received much attention due to their physico-chemical properties, biodegradability, and biocompatibility that make them good candidates for industrial and medical applications. Produced by some microorganisms PHAs accumulate within the cells of these organisms. The optimization of microbial processes to produce PHAs at a lower cost requires rapid and accurate techniques for quantification of the biopolymer in biomass. The present study describes a method based on solid-phase microextraction (SPME) coupled to gas chromatography (GC) for the determination of poly(3 hydroxybutyrate) (PHB) in Alcaligenes latus cells. First PHB was depolymerized by either methanolic or hydrolytic digestion into methyl 3-hydroxybutyrate (Me-3-HB) or crotonic acid (CA), respectively. The resulting analytes were then subjected to analysis by headspace SPME/GC with flame ionization detection (FID). The two depolymerization/SPME/GC-FID methods were optimized and applied to the analysis of PHB in bacterial biomass harvested from a fermentation process that uses A. latus. Results were compared with those obtained using GC-FID analysis of MeOH/CHCl(3) digested samples. Excellent agreement was found between the three methods but the two SPME-based methods were environmentally friendly and easier to perform. PMID- 17462662 TI - Separation of common organic and inorganic anions in atmospheric aerosols using a piperazine buffer and capillary electrophoresis. AB - The ability to monitor and quantify anionic components of aerosols is important for developing a better fundamental understanding of temporal and spatial variations in aerosol composition. Of the many methods that can be used to detect anions, capillary electrophoresis is among the most attractive ones because of its high separation efficiency, high resolving power for ionic compounds, and ability to be miniaturized for in-field monitoring. Here we present a method to baseline resolve common aerosol components nitrate, sulfate, chloride, and over two dozen organic acids in a single separation. A capillary electrophoresis separation utilizing a pH 5.78 piperazine buffer with 1,5-naphthalenedisulfonic acid as a probe for indirect UV absorbance detection was developed for this analysis. Previously, two different buffers were required to adequately separate all of these compounds. Electrophoretic mobilities, limits of detection, and migration time reproducibilities were measured for 38 organic and 8 inorganic anions. For solutions of low conductivity, detection limits for electrokinetic injections were found to be up to two orders of magnitude lower (0.2-0.4 microM) than those for pressure injection (1-45 microM). This separation was optimized and used for routine analysis of aqueous extracts of ambient atmospheric aerosols, but may be extended to other samples containing similar mixtures of anions. PMID- 17462663 TI - Direct measurement of isotopomer of intracellular metabolites using capillary electrophoresis time-of-flight mass spectrometry for efficient metabolic flux analysis. AB - We have developed a metabolic flux analysis method that is based on (13)C labeling patterns of the intracellular metabolites directly measured by capillary electrophoresis time-of-flight mass spectrometry (CE-TOFMS). The flux distribution of the central carbon metabolism in Escherichia coli was determined by this new approach and the results were compared with findings obtained by conventional GC-MS analysis based on isotopomer of the proteinogenic amino acids. There were some differences in estimation results between new approach using CE TOFMS and conventional approach using GC-MS. These were thought to be attributable to variations in measured mass distributions between amino acids and the corresponding precursors and to differences in the sensitivity of the exchange coefficients to mass distributions. However, our CE-TOFMS method facilitates high-throughput flux analysis without requiring complicated sample preparation such as hydrolysis of proteins and derivatization of amino acids. PMID- 17462664 TI - Chromatographic classification and comparison of commercially available reversed phase liquid chromatographic columns containing phenyl moieties using principal component analysis. AB - Twenty-one commercially available phenyl type RPLC packing materials have been characterized in terms of their surface coverage, hydrophobic selectivity, shape selectivity, hydrogen bonding capacity, ion exchange capacity at pH 2.7 and 7.6 and aromatic selectivity (i.e. pi-pi interaction). The phases have been compared to their corresponding C-alkyl phases, three pentafluorophenyl phases and a series of experimental phenyl phases of known bonding chemistry. Principal component analysis has been used to provide a graphical comparison of the differences and similarities between the phases. The phase's aromatic selectivity was found to be dependent on the length of the alkyl spacer between the silicon atom and the phenyl ring. PMID- 17462665 TI - Emulsification in turbulent flow 1. Mean and maximum drop diameters in inertial and viscous regimes. AB - Systematic experimental study of the effects of several factors on the mean and maximum drop sizes during emulsification in turbulent flow is performed. These factors include: (1) rate of energy dissipation, epsilon; (2) interfacial tension, sigma; (3) viscosity of the oil phase, eta(D); (4) viscosity of the aqueous phase, eta(C); and (5) oil volume fraction, Phi. The emulsions are prepared by using the so-called "narrow-gap homogenizer" working in turbulent regime of emulsification. The experiments are performed at high surfactant concentration to avoid the effect of drop-drop coalescence. For emulsions prepared in the inertial turbulent regime, the mean and the maximum drop sizes increase with the increase of eta(D) and sigma, and with the decrease of epsilon. In contrast, Phi and eta(C) affect only slightly the mean and the maximum drop sizes in this regime of emulsification. These results are described very well by a theoretical expression proposed by Davies [Chem. Eng. Sci. 40 (1985) 839], which accounts for the effects of the drop capillary pressure and the viscous dissipation inside the breaking drops. The polydispersity of the emulsions prepared in the inertial regime of emulsification does not depend significantly on sigma and epsilon. However, the emulsion polydispersity increases significantly with the increase of oil viscosity, eta(D). The experiments showed also that the inertial turbulent regime is inappropriate for emulsification of oils with viscosity above ca. 500 mPa s, if drops of micrometer size are to be obtained. The transition from inertial to viscous turbulent regime of emulsification was accomplished by a moderate increase of the viscosity of the aqueous phase (above 5 mPa s in the studied systems) and/or by increase of the oil volume fraction, Phi>0.6. Remarkably, emulsions with drops of micrometer size are easily formed in the viscous turbulent regime of emulsification, even for oils with viscosity as high as 10,000 mPa s. In this regime, the mean drop size rapidly decreases with the increase of eta(C) and Phi (along with the effects of epsilon, sigma, and eta(D), which are qualitatively similar in the inertial and viscous regimes of emulsification). The experimental results are theoretically described and discussed by using expressions from the literature and their modifications (proposed in the current study). PMID- 17462666 TI - Current status and prospects on microbial control in Japan. AB - Historically in Japan, studies on the diseases of the silkworm, Bombyx mori, as a factor affecting the well-being of the silk industry, have dominated insect pathology. However, work by Hidaka in 1933 demonstrated the possibility of controlling the pine moth, Dendrolimus spectabilis, with the fungus Beauveria bassiana and since then, various attempts have been made to develop a method to control insect pests using insect pathogens. The cypovirus product, Matsukemin, was the first microbial control product to be registered in 1974, and inactive and live Bacillus thuringiensis products were also registered and put on the market as pesticides in 1981 and 1982, respectively. Currently, there are 25 microbial insecticides on the market that constitute slightly less than 2% of all insecticides used in Japan. Adoption of biopesticides is likely to increase in the near future due to scientific advances and several new government policies that encourage the use of alternative pest control products. PMID- 17462667 TI - Acute peristome edema disease in juvenile and adult sea cucumbers Apostichopus japonicus (Selenka) reared in North China. AB - Acute peristome edema disease (APED) is a new disease that broke out in cultured sea cucumber along the Shangdong and Liaoning province coasts in China, PR, and has caused a great deal of death in Apostichopus japonicus (Selenka) since 2004. Here we report virus-like particles found in intestine epithelium of sea cucumbers reared in North China. It is the first time that sea cucumbers are reported to be infected by virus. Histological examinations showed that the viral inclusion bodies existed in intestine epithelium cells. Electron microscopic examinations show that the virions were spherical, 80-100nm in diameter, and composed of a helical nucleocapsid within an envelope with surface projections. Detailed studies on the morphogenesis of these viruses found many characteristics previously described for coronaviruses. Virus particles always congregated, and formed a virus vesicle with an encircling membrane. The most obvious cellular pathologic feature is large granular areas of cytoplasm, relatively devoid of organelles. Tubular structures within virus-containing vesicles, nucleocapsid inclusions, and double-membrane vesicles are also found in the cytopathic cells. No rickettsia, chlamydia, bacteria, or other parasitic organisms were found. PMID- 17462668 TI - A scFv antibody mutant isolated in a genetic screen for improved export via the twin arginine transporter pathway exhibits faster folding. AB - Proteins destined for export across the cytoplasmic membrane via the post translational Sec-dependent route have to be maintained in a largely unfolded state within the cytoplasm. In sharp contrast, only proteins that have folded into a native-like state within the cytoplasm are competent for export via the twin arginine translocation (Tat) pathway. Proteins that contain disulfide bonds, such as scFv antibody fragments, can be translocated via Tat only when expressed in Escherichia coli trxB gor mutant strains having an oxidizing cytoplasm. However, export is poor with the majority of the protein accumulating in the cytoplasm and only a fraction exported to the periplasmic space. Using a high throughput fluorescence screen, we isolated a mutant of the anti-digoxin 26-10 scFv from a large library of random mutants that is exported with a higher yield into the periplasm. In vitro refolding experiments revealed that the mutant scFv exhibits a 250% increase in the rate constant of the critical second phase of folding. This result suggests that Tat export competence is related to the protein folding rate and could be exploited for the isolation of faster folding protein mutants. PMID- 17462669 TI - Paxillin and ponsin interact in nascent costameres of muscle cells. AB - Muscle differentiation requires the transition from motile myoblasts to sessile myotubes and the assembly of a highly regular contractile apparatus. This striking cytoskeletal remodelling is coordinated with a transformation of focal adhesion-like cell-matrix contacts into costameres. To assess mechanisms underlying this differentiation process, we searched for muscle specific-binding partners of paxillin. We identified an interaction of paxillin with the vinexin adaptor protein family member ponsin in nascent costameres during muscle differentiation, which is mediated by an interaction of the second src homology domain 3 (SH3) domain of ponsin with the proline-rich region of paxillin. To understand the molecular basis of this interaction, we determined the structure of this SH3 domain at 0.83 A resolution, as well as its complex with the paxillin binding peptide at 1.63 A resolution. Upon binding, the paxillin peptide adopts a polyproline-II helix conformation in the complex. Contrary to the charged SH3 binding interface, the peptide contains only non-polar residues and for the first time such an interaction was observed structurally in SH3 domains. Fluorescence titration confirmed the ponsin/paxillin interaction, characterising it further by a weak binding affinity. Transfection experiments revealed further characteristics of ponsin functions in muscle cells: All three SH3 domains in the C terminus of ponsin appeared to synergise in targeting the protein to force transducing structures. The overexpression of ponsin resulted in altered muscle cell-matrix contact morphology, suggesting its involvement in the establishment of mature costameres. Further evidence for the role of ponsin in the maintenance of mature mechanotransduction sites in cardiomyocytes comes from the observation that ponsin expression was down-regulated in end-stage failing hearts, and that this effect was reverted upon mechanical unloading. These results provide new insights in how low affinity protein-protein interactions may contribute to a fine tuning of cytoskeletal remodelling processes during muscle differentiation and in adult cardiomyocytes. PMID- 17462670 TI - Mitochondria, metabolic disturbances, oxidative stress and the kynurenine system, with focus on neurodegenerative disorders. AB - The mitochondria have several important functions in the cell. A mitochondrial dysfunction causes an abatement in ATP production, oxidative damage and the induction of apoptosis, all of which are involved in the pathogenesis of numerous disorders. This review focuses on mitochondrial dysfunctions and discusses their consequences and potential roles in the pathomechanism of neurodegenerative disorders. Other pathogenetic factors are also briefly surveyed. The second part of the review deals with the kynurenine metabolic pathway, its alterations and their potential association with cellular energy impairment in certain neurodegenerative diseases. During energy production, most of the O(2) consumed by the mitochondria is reduced fully to water, but 1-2% of the O(2) is reduced incompletely to give the superoxide anion (O(2)(-)). If the function of one or more respiratory chain complexes is impaired for any reason, the enhanced production of free radicals further worsens the mitochondrial function by causing oxidative damage to macromolecules, and by opening the mitochondrial permeability transition pores thereby inducing apoptosis. These high-conductance pores offer a pathway which can open in response to certain stimuli, leading to the induction of the cells' own suicide program. This program plays an essential role in regulating growth and development, in the differentiation of immune cells, and in the elimination of abnormal cells from the organism. Both failure and exaggeration of apoptosis in a human body can lead to disease. The increasing amount of superoxide anions can react with nitric oxide to yield the highly toxic peroxynitrite anion, which can destroy cellular macromolecules. The roles of oxidative, nitrative and nitrosative damage are discussed. Senescence is accompanied by a higher degree of reactive oxygen species production, and by diminished functions of the endoplasmic reticulum and the proteasome system, which are responsible for maintenance of the normal protein homeostasis of the cell. In the event of a dysfunction of the endoplasmic reticulum, unfolded proteins aggregate in it, forming potentially toxic deposits which tend to be resistant to degradation. Cells possess adaptive mechanisms with which to avoid the accumulation of incorrectly folded proteins. These involve molecular chaperones that fold proteins correctly, and the ubiquitin proteasome system which degrades misfolded, unwanted proteins. Both the endoplasmic reticulum and the ubiquitin proteasome system fulfill cellular protein quality control functions. The kynurenine system: Tryptophan is metabolized via several pathways, the main one being the kynurenine pathway. A central compound of the pathway is kynurenine (KYN), which can be metabolized in two separate ways: one branch furnishing kynurenic acid, and the other 3-hydroxykynurenine and quinolinic acid, the precursors of NAD. An important feature of kynurenic acid is the fact that it is one of the few known endogenous excitatory amino acid receptor blockers with a broad spectrum of antagonistic properties in supraphysiological concentrations. One of its recently confirmed sites of action is the alpha7-nicotinic acetylcholine receptor and interestingly, a more recently identified one is a higher affinity positive modulatory binding site at the AMPA receptor. Kynurenic acid has proven to be neuroprotective in several experimental settings. On the other hand, quinolinic acid is a specific agonist at the N-methyl-d-aspartate receptors, and a potent neurotoxin with an additional and marked free radical producing property. There are a number of neurodegenerative disorders whose pathogenesis has been demonstrated to involve multiple imbalances of the kynurenine pathway metabolism. These changes may disturb normal brain function and can add to the pathomechanisms of the diseases. In certain disorders, there is a quinolinic acid overproduction, while in others the alterations in brain kynurenic acid levels are more pronounced. A more precise knowledge of these alterations yields a basis for getting better therapeutic possibilities. The last part of the review discusses metabolic disturbances and changes in the kynurenine metabolic pathway in Parkinson's, Alzheimer's and Huntington's diseases. PMID- 17462671 TI - Variations in the coding and regulatory sequences of the angiogenin (ANG) gene are not associated to ALS (amyotrophic lateral sclerosis) in the Italian population. AB - Potentially causative missense variations in the ANG gene and a positive association with the synonymous rs11701-G substitution was detected mainly in Irish and Scottish ALS patients. We screened 262 Italian SOD1 negative ALS patients (250 sporadic) and 415 matched controls for sequence variations in the coding, 3'/5' UTR and 5' flanking (642 bp) regions of the ANG gene. We identified 53 sequence variations of which 46 new, 20 with a minor allele frequency (MAF) >or=0.01 and only three localised in the coding sequence, namely the missense I46V, identified in one patient and two controls, and the synonymous G86G and T97T corresponding to rs11701 and rs2228653. None of the detected SNPs or of their haplotypic combinations was significantly associated with ALS susceptibility or clinical features. In conclusion, we did not detect the association with rs11701-G or with any other newly detected variation in the ANG regulatory region. Furthermore we did not identify potentially causal mutations in the coding region. PMID- 17462672 TI - Bias in the gradient-sensing response of chemotactic cells. AB - We apply linear stability theory and perform perturbation studies to better characterize, and to generate new experimental predictions from, a model of chemotactic gradient sensing in eukaryotic cells. The model uses reaction diffusion equations to describe 3(') phosphoinositide signaling and its regulation at the plasma membrane. It demonstrates a range of possible gradient sensing mechanisms and captures such characteristic behaviors as strong polarization in response to static gradients, adaptation to differing mean levels of stimulus, and plasticity in response to changing gradients. An analysis of the stability of polarized steady-state solutions indicates that the model is most sensitive to off-axis perturbations. This biased sensitivity is also reflected in responses to localized external stimuli, and leads to a clear experimental prediction, namely, that a cell which is polarized in a background gradient will be most sensitive to transient point-source stimuli lying within a range of angles that are oblique with respect to the polarization axis. Stimuli at angles below this range will elicit responses whose directions overshoot the stimulus angle, while responses to stimuli applied at larger angles will undershoot the stimulus angle. We argue that such a bias is likely to be a general feature of gradient sensing in highly motile cells, particularly if they are optimized to respond to small gradients. Finally, an angular bias in gradient sensing might lead to preferred turn angles and zigzag movements of cells moving up chemotactic gradients, as has been noted under certain experimental conditions. PMID- 17462673 TI - The different limits of weak selection and the evolutionary dynamics of finite populations. AB - Evolutionary theory often resorts to weak selection, where different individuals have very similar fitness. Here, we relate two ways to introduce weak selection. The first considers evolutionary games described by payoff matrices with similar entries. This approach has recently attracted a lot of interest in the context of evolutionary game dynamics in finite populations. The second way to introduce weak selection is based on small distances in phenotype space and is a standard approach in kin-selection theory. Whereas both frameworks are interchangeable for constant fitness, frequency-dependent selection shows significant differences between them. We point out the difference between both limits of weak selection and discuss the condition under which the differences vanish. It turns out that this condition is fulfilled by the popular parametrization of the prisoner's dilemma in benefits and costs. However, for general payoff matrices differences between the two frameworks prevail. PMID- 17462674 TI - Mechanisms of calcium sequestration during facilitation at active zones of an amphibian neuromuscular junction. AB - The calcium transients (Delta[Ca(2+)](i)) at active zones of amphibian (Bufo marinus) motor-nerve terminals that accompany impulses, visualized using a low affinity calcium indicator injected into the terminal, are described and the pathways of subsequent sequestration of the residual calcium determined, allowing development of a quantitative model of the sequestering processes. Blocking the endoplasmic reticulum calcium pump with thapsigargin did not affect Delta[Ca(2+)](i) for a single impulse but increased its amplitude during short trains. Blocking the uptake of calcium by mitochondria with CCCP had little effect on Delta[Ca(2+)](i) of a single impulse but greatly increased its amplitude during short trains. This present compartmental model is compatible with our previous Monte Carlo diffusion model of Ca(2+) sequestration during facilitation [Bennett, M.R., Farnell, L., Gibson, W.G., 2004. The facilitated probability of quantal secretion within an array of calcium channels of an active zone at the amphibian neuromuscular junction. Biophys. J. 86(5), 2674-2690], with the single plasmalemma pump in that model now replaced by separate pumps for the plasmalemma and endoplasmic reticulum, as well as the introduction of a mitochondrial uniporter. PMID- 17462675 TI - On Haldane's formulation of Luria and Delbruck's mutation model. AB - Two formulations of Luria and Delbruck's mutation model have been in common use since the 1940s. While mathematicians focused their attention on the formulation of Lea and Coulson that assumes asynchronous cell growth, biologists found more appealing the formulation of Haldane that assumes synchronous cell growth. This article attempts to solve several outstanding issues for the latter formulation. First, it provides an exact, closed-form expression for the mutant distribution by correcting a minor error in the literature. Second, it presents a novel algorithm for computing the mutant distribution, which leads to novel methods for computing point and interval estimates of mutation rates based on the maximum likelihood principle. Third, it critically examines existing methods based on the mean number of mutants. Finally, it compares the two formulations to underline their strengths and shortcomings. PMID- 17462676 TI - Mathematical modeling of the malignancy of cancer using graph evolution. AB - We report a novel computational method based on graph evolution process to model the malignancy of brain cancer called glioma. In this work, we analyze the phases that a graph passes through during its evolution and demonstrate strong relation between the malignancy of cancer and the phase of its graph. From the photomicrographs of tissues, which are diagnosed as normal, low-grade cancerous and high-grade cancerous, we construct cell-graphs based on the locations of cells; we probabilistically generate an edge between every pair of cells depending on the Euclidean distance between them. For a cell-graph, we extract connectivity information including the properties of its connected components in order to analyze the phase of the cell-graph. Working with brain tissue samples surgically removed from 12 patients, we demonstrate that cell-graphs generated for different tissue types evolve differently and that they exhibit different phase properties, which distinguish a tissue type from another. PMID- 17462677 TI - Glycine-induced long-term synaptic potentiation is mediated by the glycine transporter GLYT1. AB - The negative symptoms of schizophrenia are reverted by treatment with glycine or other agonists of the glycine-B site which facilitate NMDA receptor function. On the other hand, there are experimental observations showing that exogenous application of glycine (0.5-10mM) results in a long-lasting potentiation of glutamatergic synaptic transmission (LTP-GLY). The characterization of the mechanisms underlying LTP-GLY could be useful to develop new therapies for schizophrenia. The main goal of this work is to deepen the understanding of this potentiation phenomenon. The present study demonstrates in rat hippocampal slices that superfusion of glycine 1mM during 30 min produces a potentiation of excitatory postsynaptic potentials in CA3-CA1 pathway lasting at least 1h. Glycine application does not modify neither presynaptic fiber volley nor paired pulse facilitation of synaptic potentials. This LTP-GLY is independent of both strychnine-sensitive glycine receptors and nifedipine-sensitive calcium channels. Interestingly, LTP-GLY is not inhibited but strengthened by NMDA receptors antagonists such as AP-5 or MK-801. In contrast, LTP-GLY is partially or totally blocked with the antagonists of glycine transporter GLYT1, sarcosine or ALX-5407, respectively. These results indicate that LTP-GLY requires the activation of GLYT1, a glycine transporter co-localized and associated to NMDA receptors. In addition, the fact that NMDA receptor inhibition increases LTP-GLY magnitude, opens the possibility that these receptors could have a negative control on GLYT1 activity. PMID- 17462678 TI - Visual elements of subjective preference modulate amygdala activation. AB - What are the basic visual cues that determine our preference towards mundane everyday objects? We previously showed that a highly potent cue is the nature of the object's contour: people generally like objects with a curved contour compared with objects that have pointed features and a sharp-angled contour. This bias is hypothesized here to stem from an implicit perception of potential threat conveyed by sharp elements. Using human neuroimaging to test this hypothesis, we report that the amygdala, a brain structure that is involved in fear processing and has been shown to exhibit activation level that is proportional to arousal in general, is significantly more active for everyday sharp objects (e.g., a sofa with sharp corners) compared with their curved contour counterparts. Therefore, our results indicate that a preference bias towards a visual object can be induced by low-level perceptual properties, independent of semantic meaning, via visual elements that on some level could be associated with threat. We further present behavioral results that provide initial support for the link between the sharpness of the contour and threat perception. Our brains might be organized to extract these basic contour elements rapidly for deriving an early warning signal in the presence of potential danger. PMID- 17462679 TI - Flavonol tetraglycosides and other constituents from leaves of Styphnolobium japonicum (Leguminosae) and related taxa. AB - Two flavonol tetraglycosides comprising a trisaccharide at C-3 and a monosaccharide at C-7 were isolated from the leaves of Styphnolobium japonicum (L.) Schott and characterised as the 3-O-alpha-rhamnopyranosyl(1-->2)[alpha rhamnopyranosyl(1-->6)]-beta-glucopyranoside-7-O-alpha-rhamnopyranosides of quercetin and kaempferol. The 3-O-alpha-rhamnopyranosyl(1-->2)[alpha rhamnopyranosyl(1-->6)]-beta-galactopyranoside-7-O-alpha-rhamnopyranoside of kaempferol, the 3-O-alpha-rhamnopyranosyl(1-->2)[alpha-rhamnopyranosyl(1-->6)] beta-glucopyranosides of kaempferol and quercetin and the 3-O-alpha rhamnopyranosyl(1-->2)[alpha-rhamnopyranosyl(1-->6)]-beta-galactopyranoside of kaempferol were also obtained from this species for the first time. Some or all of these flavonol tetra- and triglycosides were detected in 17 of 18 specimens of S. japonicum examined from living and herbarium material, although the most abundant flavonoid in the leaves was generally quercetin 3-O-alpha rhamnopyranosyl(1-->6)-beta-glucopyranoside (rutin). The triglycosides, but not the tetraglycosides, were detected in herbarium specimens of Styphnolobium burseroides M. Sousa, Rudd & Medrano and Styphnolobium monteviridis M. Sousa & Rudd, but specimens of Styphnolobium affine (Torrey & A. Gray) Walp. contained a different profile of flavonol glycosides. The flavonol tetra- and triglycosides of S. japonicum were also present in leaves of Cladrastis kentukea (Dum. Cours.) Rudd, a representative of a genus placed close to Styphnolobium in current molecular phylogenies. An additional constituent obtained from leaves of Styphnolobium japonicum was identified as the maltol derivative, 3-hydroxy-2 methyl-4H-pyran-4-one 3-O-(4'-O-p-coumaroyl-6'-O-(3-hydroxy-3-methylglutaroyl)) beta-glucopyranoside. PMID- 17462680 TI - Missed opportunities? Locating health promotion within multidisciplinary public health. AB - This paper examines the complex interconnections between the development of health promotion and multidisciplinary public health, respectively. Health promotion takes a distinctive interdisciplinary and multiprofessional perspective on health. Historically, it has brought together practitioners from varied disciplinary backgrounds, education and training. It therefore brings real advantages to the public health enterprise, where the goal is to bridge organisations, professions and partners to collectively address key determinants of health in the most effective manner. This paper debates the contribution health promotion has made to the development of multidisciplinary public health over the past 30 years and explores the principles, values, professional bases and practices of both. It is argued that health promotion's contribution to the development of 'the new public health' was critical, while its status and role within multidisciplinary public health remain problematic and unresolved. The nature of these dilemmas is discussed, reflecting on missed opportunities and possible resolutions. PMID- 17462681 TI - Integrating services for public health: challenges facing multidisciplinary partnership working. PMID- 17462682 TI - Fringe benefits? The impact of the Multidisciplinary Public Health Forum in the UK outside of England. AB - The impact of the UK Multidisciplinary Public Health Forum in developing the public health workforce has been the subject of some discussion. However, much of the work to date has focused on the impact of the Forum in the English context. This paper uses the same techniques used to examine the impact of the Forum in England to describe the situation in Scotland. The key themes that emerge are that the impact of the Multidisciplinary Public Health Forum in Scotland was marginal and that a multi-professional approach was the more likely route of development. The adoption of regulatory policy by the UK Government in Westminster had the effect of producing a multidisciplinary approach in Scotland. The relationship of these findings to the experiences of Wales and Northern Ireland is discussed. PMID- 17462683 TI - The role of the Faculty of Public Health (Medicine) in developing a multidisciplinary public health profession in the UK. AB - The Faculty of Public Health (FPH) started out its life as the Faculty of Community Medicine. Its initial membership was drawn from the ranks of Medical Officers of Health (MOsH), medical administrators and consultants working in communicable disease. Born under the aegis of the three Royal Colleges of Physicians, it was de facto for members of the medical profession. This was despite the intention of some of the prime movers in its creation that its membership should reflect the multidisciplinary nature of specialist public health. As such, whilst the Faculty's establishment was indeed a triumph, the triumph was only partial, since many senior public health professionals were precluded from full membership. Over the years this situation has changed, but the road to the Faculty's current open policy, based on achieving public health excellence rather than holding a professional badge, has not been a smooth one. The fears of many medical members that the specialty would be down graded through opening up its membership posed successive presidents with many, often justifiable, challenges. In this article we, former presidents, reflect on the key events during our successive tenures. PMID- 17462684 TI - [218 tooth extraction in patients taking platelet aggregation inhibitors]. AB - INTRODUCTION: The aim of this survey was to show that tooth extraction could be performed in patients taking antiplatelet agents. The main indication of antiplatelet agents is to reduce the thrombotic disease. MATERIAL AND METHOD: The authors made a descriptive and retrospective analysis of 52 patients taking antiplatelet agents in their department between February 2003 and January 2005. Two hundred and eighteen tooth extractions were performed. For each extraction, a protocol of local hemostasis (filling, suture, compression) was applied. RESULTS: Three hemorrhagic sockets were reported out of 218 extractions performed without stopping the antiplatelet agent treatment (1.3%). One patient presented with persistent bleeding out of 52 cases (1.9%). No hemostasis had been performed on this patient; a local hemostasis was performed during surgical revision, which stopped the persistent bleeding. DISCUSSION: These results show that the hemorrhagic risk can be controlled by a local hemostasis protocol. PMID- 17462685 TI - Involvement of CYP 2E1 enzyme in ovotoxicity caused by 4-vinylcyclohexene and its metabolites. AB - 4-Vinylcyclohexene (VCH) is bioactivated by hepatic CYP 2A and 2B to a monoepoxide (VCM) and subsequently to an ovotoxic diepoxide metabolite (VCD). Studies suggest that the ovary can directly bioactivate VCH via CYP 2E1. The current study was designed to evaluate the role of ovarian CYP 2E1 in VCM-induced ovotoxicity. Postnatal day 4 B6C3F(1) and CYP 2E1 wild-type (+/+) and null (-/-) mouse ovaries were cultured (15 days) with VCD (30 microM), 1,2-VCM (125-1000 microM), or vehicle. Twenty-eight days female CYP 2E1 +/+ and -/- mice were dosed daily (15 days; ip) with VCH, 1,2-VCM, VCD or vehicle. Following culture or in vivo dosing, ovaries were histologically evaluated. In culture, VCD decreased (p<0.05) primordial and primary follicles in ovaries from all three groups of mice. 1,2-VCM decreased (p<0.05) primordial follicles in B6C3F(1) and CYP 2E1 +/+ ovaries, but not in CYP 2E1 -/- ovaries in culture. 1,2-VCM did not affect primary follicles in any group of mouse ovaries. Conversely, following in vivo dosing, primordial and primary follicles were reduced (p<0.05) by VCD and VCM in CYP2E1 +/+ and -/-, and by VCH in +/+ mice. The data demonstrate that, whereas in vitro ovarian bioactivation of VCM requires CYP 2E1 enzyme, in vivo CYP 2E1 plays a minimal role. Thus, the findings support that hepatic metabolism dominates the contribution made by the ovary in bioactivation of VCM to its ovotoxic metabolite, VCD. This study also demonstrates the use of a novel ovarian culture system to evaluate ovary-specific metabolism of xenobiotics. PMID- 17462686 TI - Skin: major target organ of allergic reactions to small molecular weight compounds. AB - Skin is a major target organ for allergic reactions to small molecular weight compounds. Drug allergic reactions may be life-threatening such as in the case of anaphylactic reactions or bullous drug reactions and occur in about 5% of all hospitalized patients. Allergic contact dermatitis has an enormous influence on the social life of the patient because it is the most frequent reason for occupational skin diseases and the treatment and prevention of this disease cost approximately euro 3 billion per year in Germany. The different proposed pathophysiological pathways leading to a drug eruption are discussed in this paper. All major enzymes which are involved in the metabolism of xenobiotica were shown to be present in skin. Evidence supporting the role of metabolism in the development of drug allergy and allergic contact dermatitis is demonstrated in the example of sulphonamides and fragrances. PMID- 17462687 TI - Poisoning by Cnestis ferruginea in Casamance (Senegal): an etiological approach. AB - Since several years, in the area of Kabrousse in Casamance (Senegal), a neurotoxic syndrome has caused more than 50 human deaths. Field studies showed that epidemic could be due to consumption of leave decoction of Cnestis ferruginea, a tropical plant belonging to the Connaraceae family. An ethnobotanical study has been conducted in order to investigate the traditional uses of C. ferruginea, and describe the circumstances and the symptoms of this plant poisoning. As a first experimental approach, the leave decoction was tested for its ability to induce cytotoxic effects using the XTT method. A phytochemical approach revealed the presence of methionine sulfoximine (MSX), a neurotoxic amino acid, in the plant extract by gas chromatography-mass spectrometry (GC-MS). The description of this poisoning, the cytotoxic activity of the decoction and the occurence of MSX in leaves of C. ferruginea constituted the first etiological data on this poisoning. PMID- 17462688 TI - Use of ultrasound for the determination of flour quality. AB - Within the baking industry, the control of dough properties is required to achieve final product quality and consistency. Traditional methods for dough testing are slow and off-line and do not provide fundamental rheological information. There is therefore a need for the development of fast and on-line instruments capable of providing relevant data for baking. Ultrasonics provide a non-destructive, rapid and low cost technique for the measurement of physical food characteristics. In this work, the water content of dough is investigated using ultrasonic techniques. The capability of ultrasound measurements for discriminating flours for different purposes is also studied. Doughs from more than 30 flours were characterised rheologically using a Chopin Alveograph and a Brabender Extensograph. Ultrasound measurements on the doughs prepared from these flours were also performed. The measurements were correlated, showing that ultrasound was an alternative measurement method to discriminate types of flours for different purposes. PMID- 17462689 TI - Fundamental study on ultrasonic measurement system to detect penetration of boulders using cepstrum analysis. AB - Generally, ultrasonic method and an elastic impact wave method are mentioned as examples representing the nondestructive test method. However, the ultrasonic method is known for the investigation of shallow depth becoming very shallow because of its small input. And the elastic impact wave method uses for example, a hammer for the discrimination of the reflection wave produced by minute cracks existing in the object, which is difficult. Theoretically, if it becomes possible to input the vibration of an ultrasonic domain into artificial structures such as a concrete and a boulder with strong amplitude, deeper investigation of depth and higher precision will be attained. The authors have developed a new ultrasonic method by using ultrasonic wave of the chirp signal, using the input of this system and using auto correlation and cepstrum as the analysis methods. In this paper, the experiment is classified into Pattern 1 and Pattern 2. In Pattern 1, the experiment of receiving the reflection wave was carried out using the concrete block with a width of 1300 mm and the down chirp signal from 50 kHz to 40 kHz. In Pattern 1, a reflection wave was clearly detected, and the length of the concrete block was calculated using auto correlation analysis, and was obtained as 1311.76 mm. In Pattern 2, the experiment of receiving the reflection wave was carried out using the same concrete block and the down chirp signal from 150 kHz to 130 kHz. In Pattern 2, the direct wave and the reflection wave were mixed, therefore it was not possible to presume the length from the obtained waveform. So cepstrum analysis method was used, and the traveling time of the reflection wave was obtained. In this result, the length of the concrete block was given as 1278.25 mm. PMID- 17462690 TI - Numerical predictions and experiments for optimizing hidden corrosion detection in aircraft structures using Lamb modes. AB - To increase the sensitivity of Lamb waves to hidden corrosion in aircraft structures, a preliminary step is to understand the phenomena governing this interaction. A hybrid model combining a finite element approach and a modal decomposition method is used to investigate the interaction of Lamb modes with corrosion pits. The finite element mesh is used to describe the region surrounding the corrosion pits while the modal decomposition method permits to determine the waves reflected and transmitted by the damaged area. Simulations make easier the interpretation of some parts of the measured waveform corresponding to superposition of waves diffracted by the corroded area. Numerical results permit to extract significant information from the transmitted waveform and thus to optimize the signal processing for the detection of corrosion at an early stage. Now, we are able to detect corrosion pits down to 80 mum depth distributed randomly on a square centimeter of an aluminum plate. Moreover, thickness variations present on aircraft structures can be discriminated from a slightly corroded area. Finally, using this experimental setup, aircraft structures have been tested. PMID- 17462691 TI - Calcium enhances mouse keratinocyte differentiation in vitro to differentially regulate expression of papillomavirus authentic and codon modified L1 genes. AB - Here, we first wished to establish for mouse primary keratinocytes (KCs) the Ca(2+) concentrations that were associated with KC differentiation in vitro. Using the range of Ca(2+) concentrations (0-6 mM) to differentiate primary KCs in culture to varying extents for 2 days, we then examined how KC differentiation impacted on expression of papillomavirus (PV) native (Nat) and codon modified (Mod) L1 genes. L1 mRNAs transcribed from either Nat or Mod L1 genes were present in similar amounts in KCs exposed to six Ca(2+) concentrations. However, expression of the L1 proteins from two Mod L1 genes were down-regulated, with no L1 signal detected in KCs exposed to 6 mM Ca(2+). In contrast, L1 proteins expressed from the two Nat L1 genes were not detectable in KCs without Ca(2+), but dramatically up-regulated as the KC cultures exposed to Ca(2+) from 0.5 to 2 mM, then down-regulated in KCs exposed to Ca(2+) from 4 to 6 mM. The different regulatory roles of the Ca(2+) in L1 protein expression from Nat and Mod L1 genes in cultured KCs were confirmed by TGF-beta1 experiments. We observed that aminoacyl-tRNAs (aa-tRNAs) from the 2 mM Ca(2+)-treated KCs only significantly enhanced the Nat L1 mRNAs translation in vitro, suggesting that aa-tRNAs play a differentially regulatory role in translations of the PV Nat and Mod L1 mRNAs. Importantly, the Ca(2+) experimental model provides evidence that mouse primary KCs could be transiently infected by BPV1 virus to express L1 mRNA and protein, which is very useful for future HPV virus infection study. PMID- 17462692 TI - Myristoylation signal transfer from the large to the middle or the small HBV envelope protein leads to a loss of HDV particles infectivity. AB - A myristate linked to the N-terminus of the large hepatitis B virus (HBV) envelope protein was found to be required for infectivity of the hepatitis delta virus (HDV). Myristoylation of the large HBV envelope protein being known as indispensable for HBV infectivity, this result further demonstrates the similarities between the HBV and HDV entry pathways. In addition, the transfer of the N-myristoylation signal from the large to the middle or the small HBV envelope protein led in both cases to a loss of HDV infectivity. Hence, it is suggested that viral entry could depend on a physical link, or a spatial association, between the N-terminal receptor-binding polypeptide of the large protein and the myristoyl anchor linked to glycine-2. PMID- 17462693 TI - The role of the downstream signal sequences in the maturation of the HBV middle surface glycoprotein: development of a novel therapeutic vaccine candidate. AB - The signal sequences that mediate entry of the hepatitis B virus (HBV) envelope proteins into the endoplasmic reticulum (ER) are located within the S domain at positions 11-32 and at positions 80-98 (from the start of the S domain). In addition, hydrophobic patches at positions 160-184 and 189-210 of the S domain may also be involved in entry into the ER. The role of each of these domains in the entry of the HBV M glycoprotein into the ER was studied by deletion mutations of each of the signal sequences. Glycosylation of proteins was used as a marker of entry into the ER. Our results indicate that association with the ER could not be prevented by the deletion of either individual or combinations of the HBV signal sequences. M protein lacking signal sequence I was able to enter the ER and had limited secretion. In contrast, M protein lacking signal sequence II could not be secreted but still entered the ER. M protein lacking signal sequences I and II, while still associated with the ER, was rapidly degraded by the cytosolic proteasome. The potential use of such a vector as a CTL vaccine was tested through an in vitro antigen presentation assay. In this assay, a DNA vaccine candidate lacking signal sequences I and II lead to a >6-fold increase in CTL activation, as compared to the vector expressing wild type M protein. These results suggest that increased degradation of the HBV envelope proteins can lead to enhanced antigen presentation. PMID- 17462694 TI - Variola virus topoisomerase: DNA cleavage specificity and distribution of sites in Poxvirus genomes. AB - Topoisomerase enzymes regulate superhelical tension in DNA resulting from transcription, replication, repair, and other molecular transactions. Poxviruses encode an unusual type IB topoisomerase that acts only at conserved DNA sequences containing the core pentanucleotide 5'-(T/C)CCTT-3'. In X-ray structures of the variola virus topoisomerase bound to DNA, protein-DNA contacts were found to extend beyond the core pentanucleotide, indicating that the full recognition site has not yet been fully defined in functional studies. Here we report quantitation of DNA cleavage rates for an optimized 13 bp site and for all possible single base substitutions (40 total sites), with the goals of understanding the molecular mechanism of recognition and mapping topoisomerase sites in poxvirus genome sequences. The data allow a precise definition of enzyme-DNA interactions and the energetic contributions of each. We then used the resulting "action matrix" to show that favorable topoisomerase sites are distributed all along the length of poxvirus DNA sequences, consistent with a requirement for local release of superhelical tension in constrained topological domains. In orthopox genomes, an additional central cluster of sites was also evident. A negative correlation of predicted topoisomerase sites was seen relative to early terminators, but no correlation was seen with early or late promoters. These data define the full variola virus topoisomerase recognition site and provide a new window on topoisomerase function in vivo. PMID- 17462695 TI - Abutilon mosaic virus DNA B component supports mechanical virus transmission, but does not counteract begomoviral phloem limitation in transgenic plants. AB - Different Nicotiana benthamiana lines stably transformed with Abutilon mosaic virus (AbMV) dimeric DNA B were capable of systemically spreading complete bipartite AbMV genomes, following agroinoculation of DNA A alone. Constitutively expressed viral movement protein (BC1) did not induce any persistent disease phenotype, but plants developed transient morphological abnormalities such as radially symmetric leaves after kanamycin withdrawal. Systemic AbMV infection produced symptoms and virus titers indistinguishable from those in non-transgenic plants. In systemically invaded leaves, the begomovirus remained phloem-limited, whereas the plants' susceptibility to mechanical transmission of AbMV was enhanced by a factor of three to five, as compared to non-transgenic controls. Hence, DNA B-encoded movement functions can complement local movement to the phloem after mechanical transmission, but fail to support viral invasion of non phloem cells in systemically infected organs, indicating that the phloem restriction of AbMV does not result predominantly from a lack of transport competence in mesophyll tissues. PMID- 17462696 TI - Second-order optic flow processing. AB - Optic flow-large-field rotational and radial motion-is processed as efficiently as translational motion for first-order (luminance-defined) stimuli. However, it has been suggested recently that the same pattern does not hold for second-order (e.g. contrast-defined) stimuli. We used random dot kinematogram (RDK) stimuli to determine whether global processing of optic flow is as efficient as processing of global translational motion for both first- and second-order stimuli. For first-order stimuli, we found that coherence thresholds for radial and rotational motion were equivalent to thresholds for translational motion, supporting previous findings. For second-order stimuli we found, firstly, that given sufficient contrast, second-order optic flow can be processed as efficiently as first-order optic flow and, secondly, that rotational and translational second order motion are processed with equal efficiency. This contradicts the suggestion that there is a loss of efficiency between integration of second-order global motion and second-order optic flow. The third interesting finding was that the processing of radial second-order motion appears to suffer from a deficit that is dependent upon both the contrast and spatial extent of the stimulus. Further experiments discounted the possibility that the observed deficit is caused by a centrifugal or centripetal bias, but demonstrated that a longer temporal integration period for radial second-order motion is responsible for the observed difference. For durations of approximately 850ms, all three types of motion are processed with equal efficiency. PMID- 17462697 TI - Intrinsic growth rate: a new approach to evaluate the effects of temperature, photoperiod and phosphorus-nitrogen concentrations on duckweed growth under controlled eutrophication. AB - A comprehensive model based on a new approach was developed to simulate the duckweed growth under controlled conditions. Contrary to other approaches which use the specific growth rate, this approach uses the intrinsic growth rate which permits to differentiate the effect of duckweed mat density from that of temperature, photoperiod and phosphorus-nitrogen concentrations. The model was calibrated using data from laboratory experiments carried out during the present study and validated using other data from two literature sources. In both cases, the results demonstrated that the model was capable of predicting duckweed growth with a reliability of 95%. PMID- 17462698 TI - Visualisation of transient processes in biofilms by optical coherence tomography. AB - Biofilms occur in natural and engineered water systems. In technical processes, biofouling lowers the water quality and increases the frictional resistance in tubes. In wastewater treatment plants, biofilms are used for the removal of organic and inorganic pollutants. For improvement of antifouling strategies and for process optimisation in wastewater treatment plants, analytical techniques for online monitoring of biofilms are needed. Optical coherence tomography (OCT) is a non-invasive optical tomography technique, which is increasingly applied in medical diagnostics. It reveals photon-reflecting structures in tissue with lateral and axial resolution in the range of 10 microm. In this paper, we demonstrate the capabilities of OCT for the monitoring of biofilm structures and their detachment. OCT is able to reveal spatially resolved structural information on biofilm without staining. A main focus of this work is set on the ability of OCT to monitor transient processes with temporal resolution in a second to minute scale. These key features of OCT allow online monitoring of biofilm growth and detachment in a flow channel. Three-dimensional (3D) imaging quality, spatial resolution, and temporally resolved profiling of biofilms are demonstrated. The results give rise to the hope that OCT may evolve to a standard tool for monitoring of biofilm density. PMID- 17462699 TI - Organic polyelectrolytes in water treatment. AB - The use of polymers in the production of drinking water is reviewed, with emphasis on the nature of the impurities to be removed, the mechanisms of coagulation and flocculation, and the types of polymers commonly available. There is a focus on polymers for primary coagulation, their use as coagulant aids, in the recycling of filter backwash waters, and in sludge thickening. Practicalities of polymer use are discussed, with particular attention to polymer toxicity, and the presence of residual polymer in the final drinking water. The questions of polymer degradation and the formation of disinfection by-products are also addressed. PMID- 17462700 TI - Concentrations of p,p'-DDE in plasma of nestling Wood Storks from Georgia. AB - We determined the concentrations of organochlorine pesticides and total PCBs in plasma of nestling Wood Storks (Mycteria americana) from two colonies in Georgia in 2000. Of 20 compounds analyzed for, only p,p'-DDE was quantified in reportable concentrations, ranging from <2.0-174.0 ng/g wet weight (ww). Concentrations of p,p'-DDE were significantly greater (P<0.0001) in plasma of nestlings from an inland colony, Chew Mill (x 16.9 ng/g,ww) than a coastal colony, Harris Neck (x 1.2 ng/g,ww). Concentrations from nestlings from Chew Mill were significantly different among sampling weeks (P=0.01), with week 8 being significantly greater than weeks 1, 2, and 5. While the Chew Mill colony had much greater concentrations of p,p'-DDE in plasma of nestlings than the Harris Neck colony, reproduction was greater there (1.8+/-1.0 SD fledged young per nesting attempt) than Harris Neck (1.4+/-1.0 SD fledged young per nesting attempt). While concentrations of p,p'-DDE determined in plasma of nestling Wood Storks are an indirect measure of adult exposure to environmental toxicants, concentrations reported would not be considered detrimental to reproduction in these colonies. PMID- 17462701 TI - Salting-in and salting-out effects of ionic and neutral osmotica on limonene and linalool Henry's law constants and octanol/water partition coefficients. AB - Foliar emission rates of plant-generated volatile monoterpenes depend on monoterpene partitioning between air, aqueous and lipid-phases in the leaves. While Henry's law constants (H pc, equilibrium gas/water partition coefficient) and octanol/water partition coefficients (K OW) for pure water have been previously used to simulate monoterpene emissions from the leaves, aqueous phase in plants is a complex solution of electrolytes and neutral osmotica. We studied the effects of dissociated compounds KCl and glycine and sugars glucose, sorbitol and sucrose with concentrations between 0 and 1M on H pc and K OW values for limonene and linalool. Linalool with ca. 1500-fold lower H(pc) (2.62 Pa m(3)mol( 1) for pure water at 30 degrees C) and ca. 30-fold lower K OW (955 mol mol(-1) for pure water at 25 degrees C) is the more hydrophilic compound of the two monoterpenes. H pc of both monoterpenes increased with increasing concentration of both ionic compounds and sorbitol, but decreased with increasing glucose and sucrose concentrations. The salting-out coefficients for H pc (kH) were ca. an order of magnitude larger for more hydrophilic compound linalool than for more hydrophobic limonene. For linalool, co-solutes modified H pc by 30-50% at the highest concentration (1M) tested. The effect of temperature on the salting-out coefficient of KCl was minor. As with H pc, K OW increased with increasing the concentration of KCl, glycine and sorbitol, and decreased with increasing glucose and sucrose concentrations. For limonene, co-solutes modified K OW by 20-50% at the highest concentration used. For linalool, the corresponding range was 10-35%. Salting-out coefficients for H pc and K OW were correlated, but the lipid solubility was more strongly affected than aqueous solubility in the case of limonene. Overall, these data demonstrate physiologically important effects of co solutes on H pc and K OW for hydrophilic monoterpenes and on K OW for hydrophobic monoterpenes that should be included in current emission models. PMID- 17462702 TI - Bioaccumulation of Cd by a European lacertid lizard after chronic exposure to Cd contaminated food. AB - Apart from analyses for elemental contaminants in field collected specimens, very little is known about the assimilation, accumulation and toxic effects of inorganic contaminants in reptiles. This study examined the chronic accumulation of Cd in a European lacertid lizard (Podarcis carbonelli) following dietary provision of an environmentally realistic concentration of Cd for 21 weeks. Lizards were provided with Cd that had either been biologically incorporated into crickets, or as Cd(NO3)2 added superficially to crickets just prior to feeding. Among both treatment groups Cd accumulated in tissues in the following order of concentration: gut>liver>kidney>carcass. The majority of the Cd was retained within the gut, and transfer to internal organs was low. Morphological indices, brain and plasma cholinesterase activities, gut and liver metallothionein content, and standard metabolic rate were measured as biomarkers of exposure and effect; however, no differences between control lizards and Cd-treated lizards were observed. PMID- 17462703 TI - Biological responses of wheat (Triticum aestivum) plants to the herbicide chlorotoluron in soils. AB - Chlorotoluron is a phenylurea herbicide that is widely used for controlling grass weeds in the land of cereal, cotton and fruit production. However, extensive use of this herbicide may lead to its accumulation in ecosystems, thus inducing the toxicity to crops and vegetables. To assess chlorotoluron-induced toxicity in plants, we performed the experiment focusing on the metabolic adaptation of wheat plants (Triticum aestivum) to the chlorotoluron-induced oxidative stress. The wheat plants were cultured in the soils with chlorotoluron at concentrations of 0 25mg/kg. Chlorotoluron accumulation in plants was positively correlated with the external chlorotoluron concentrations, but negatively with the plant growth. Treatment with chlorotoluron induced the accumulation of O(2)(-) and H(2)O(2) in leaves and resulted in the peroxidation of plasma membrane lipids in the plant. We measured the endogenous proline level and found that it accumulated significantly in chlorotoluron-exposed roots and leaves. To understand the biochemical responses to the herbicide, activities of the antioxidant enzymes, such as superoxide dismutase (SOD), peroxidase (POD), catalase (CAT) and ascorbate peroxidase (APX) were assayed. Analysis of SOD activity by non denaturing polyacrylamide gel electrophoresis (PAGE) revealed that there were three isoforms in the roots and leaves, but the isoforms in the tissues showed different patterns. Also, using the native PAGE, 6 isoforms of root POD and 10 in leaves were detected. The total activity of POD in roots was significantly enhanced. Activities of APX in roots and leaves showed a similar pattern. The CAT activities were generally suppressed under the chlorotoluron exposure. PMID- 17462704 TI - Advanced oxidation of natural organic matter using hydrogen peroxide and iron coated pumice particles. AB - The oxidative removal of natural organic matter (NOM) from waters using hydrogen peroxide and iron-coated pumice particles as heterogeneous catalysts was investigated. Two NOM sources were tested: humic acid solution and a natural source water. Iron coated pumice removed about half of the dissolved organic carbon (DOC) concentration at a dose of 3000 mg l(-1) in 24 h by adsorption only. Original pumice and peroxide dosed together provided UV absorbance reductions as high as 49%, mainly due to the presence of metal oxides including Al(2)O(3), Fe(2)O(3) and TiO(2) in the natural pumice, which are known to catalyze the decomposition of peroxide forming strong oxidants. Coating the original pumice particles with iron oxides significantly enhanced the removal of NOM with peroxide. A strong linear correlation was found between iron contents of coated pumices and UV absorbance reductions. Peroxide consumption also correlated with UV absorbance reduction. Control experiments proved the effective coating and the stability of iron oxide species bound on pumice surfaces. Results overall indicated that in addition to adsorptive removal of NOM by metal oxides on pumice surfaces, surface reactions between iron oxides and peroxide result in the formation of strong oxidants, probably like hydroxyl radicals, which further oxidize both adsorbed NOM and remaining NOM in solution, similar to those in Fenton-like reactions. PMID- 17462705 TI - Evaluation of the thermal/optical reflectance method for discrimination between char- and soot-EC. AB - Many optical, thermal and chemical methods exist for the measurement of elemental carbon (EC) but are unable or neglect to differentiate between the different forms of EC such as char- or soot-EC. The thermal/optical reflectance (TOR) method applies different temperatures for measuring EC and organic carbon (OC) contents through programmed, progressive heating in a controlled atmosphere, making available eight separate carbon fractions - four OC, one pyrolyzed organic carbon, and three EC. These fractions were defined by temperature protocol, oxidation atmosphere, and laser-light reflectance/transmittance. Stepwise thermal evolutional oxidation of the TOR method makes it possible to distinguish char- from soot-EC. In this study, different EC reference materials, including char and soot, were used for testing it. The thermograms of EC reference materials showed that activation energy is lower for char- than soot-EC. Low-temperature EC1 (550 degrees C in a 98% He/2% O2 atmosphere) is more abundant for char samples. Diesel and n-hexane soot samples exhibit similar EC2 (700 degrees C in a 98% He/2% O2 atmosphere) peaks, while carbon black samples peaks at both EC2 and EC3 (800 degrees C in a 98% He/2% O2 atmosphere). These results supported the use of the TOR method to discriminate between char- and soot-EC. PMID- 17462706 TI - Identification of methyl chloride-emitting plants and atmospheric measurements on a subtropical island. AB - A survey of methyl chloride (CH3Cl)-emitting plants was performed at a subtropical island in Japan (Iriomote Island). Among the 187 species of tropical/subtropical plants investigated, 33 species from a variety of families were identified as CH3Cl-emitting plants. The strongest emitters were Osmunda banksiifolia, Cibotium balometz, Angiopteris palmiformis, Vitex rotundifolia, Vitex trifolia, and Excoecaria agalloch, each with CH3Cl emission rates exceeding 1microg (gdrywt)(-1)h(-1). The first three species are ferns, and the last three are halophilous plants. Based on our results, the character of CH3Cl emission is likely to be shared at the genus level but not always at the family level. The atmospheric CH3Cl distribution measured on Iriomote Island showed significant enhancement in forested sites (up to 2750 ppt) and a higher concentration on the downwind shore than on the upwind shore. As previously reported, our findings provide strong evidence for the high emission of CH3Cl from tropical/subtropical forests. PMID- 17462707 TI - Effect of various cyclodextrins on photodegradation of a hydrophobic herbicide in aqueous suspensions of different soil colloidal components. AB - This paper investigated the photochemical behaviour of the herbicide norflurazon (NFL) in the presence of different soil colloidal components and several cyclodextrins (CDs). The interaction of NFL with CDs yielded the formation of inclusion complexes at 1:1 stoichiometric ratio in solution, with an increase of the herbicide solubility. The irradiation of NFL aqueous solutions in the presence of CDs showed that the higher the formation constant of NFL-CD complexes (Kc) and their solubility, the higher their photocatalytic effects, following the CDs in the order: RAMEB>HPBCD>beta-CD>alpha-CD>gamma-CD. The presence of the different soil colloidal components in aqueous suspension provoked the reduction of the NFL photodegradation rate, due to a screening effect, especially when goethite and humic acids were present. No disappearance of NFL was detected in parallel studies carried out in the dark, except in the case of humic acids, where a 5% adsorption of the initial amount of NFL was adsorbed in the dark control. The presence of the different CDs in such systems showed an inductive photodegradation effect on the herbicide. This could be largely explained by the inclusion effects of CDs in catalyzing interactions between NFL and certain reactive radicals generated by the different colloidal components. Although this work was carried out at laboratory scale and therefore, has limited applications, it reveals that cyclodextrins increase solubilization of hydrophobic herbicides and could lead to their increased photodegradation. This could be a promising method for pesticide-contaminated water remediation. However, it is important to consider the effect of the soil colloidal components in the different aquatic systems and their concentrations, since they can alter the photodegradative effects of the cyclodextrins. PMID- 17462708 TI - In situ immobilization of Cu(II) in soils using a new class of iron phosphate nanoparticles. AB - This study tested the feasibility of using a new class of iron phosphate (vivianite) nanoparticles synthesized using sodium carboxymethyl cellulose (NaCMC) as a stabilizer for in situ immobilization of Cu(II) in soils. Transmission electron microscopy measurements demonstrated that the particle size was about 8.4+/-2.9 nm. Batch tests showed that nano-sized vivianite particles can effectively reduce the leachability and in vitro bioaccessibility of Cu(II) in three representative soils (calcareous, neutral, and acidic) at the low doses of 0.61 and 3.01 mg PO(4) g(-1) soil. The Cu leachability was evaluated by the toxicity characteristic leaching procedure and in vitro bioaccessibility was evaluated by the physiological based extraction test. In the case of soil amendment with nanoparticles in 3.01 mg PO(4) g(-1) soil, Cu leachability reduced 63-87% and Cu concentrations in TCLP extract decreased from 1.74-13.33 mg l(-1) to 0.23-2.55 mg l(-1) after those soils were amended for 56 d. Meanwhile, the bioaccessibility of Cu was reduced by 54-69%. Sequential extraction procedures showed the significant decrease of water soluble/exchangeable Cu(II) and carbonate bound fractions and concomitant increase of Cu residual fraction after the soils were amended with the nanoparticles, suggesting that the formation of copper phosphate minerals through precipitation and adsorption was probably responsible for the decrease of Cu availability in soils. Visual MINTEQ modeling further revealed that Cu(3)(PO(4))(2) and Cu(5)(PO(4))(3)OH were formed in the vivianite-solid Cu(II) system, resulting in the decreased solubility of the Cu(II) in the acidic pH range. PMID- 17462709 TI - Electrochemical treatment of human urine for its storage and reuse as flush water. AB - We proposed the electrochemical treatment of human urine to enable its storage without the accompanying unpleasant odor. This urine can then be reused as flush water in toilets as a means to tackle water shortage problems. In laboratory scale experiments, the time-dependent variation in the pH of human urine, after the addition of urease, could be suppressed by chlorine produced via the electrochemical treatment of diluted human urine. Ureolysis was quantified by pH increase within 100 h. This suppression occurred as a result of an irreversible change in the conformation of urease that resulted in its inactivation at an oxidation-reduction potential (ORP) of ca. 240 mV or above. Due to the electrochemical inactivation of urease during the entire storage period of urine, the hydrolysis of urea in urine, which results in the production of the unpleasant odor due to ammonia formation, can be avoided. Thus, the treatment enables the storage of urine for its reuse as flush water in toilets. PMID- 17462710 TI - Environmental fate of endocrine-disrupting dimethyl phthalate esters (DMPE) under sulfate-reducing condition. AB - Dimethyl phthalate esters (DMPE) can easily be released into the environment from plastic products. As endocrine disruptors, DMPE mimic estrogenic activities in animals and humans. The metabolites of DMPE are suspected to cause even more serious health problems. Among the common sterilization techniques adopted in the study of DMPE degradation, the average loss of the parent DMPE compounds after autoclaving was as high as 21.26%. In contrast, the loss after 0.2 microm filtration was significantly lower at 2.28%. It is suggested that filtration should be used over autoclaving for sterilizing DMPE. The environmental fate of DMPE under sulfate-reducing condition was simulated and studied in microcosm system. It was observed that dimethyl phthalate (DMP), dimethyl isophthalate (DMI) and dimethyl terephthalate (DMT) could not be mineralized over an extended period of 6 months, but with the transformation to the respective monomethyl phthalate and/or phthalic acid. The dominant species of microorganisms utilizing individual DMPE isomer as the sole carbon source were isolated and identified as facultative anaerobe Thauera sp., Xanthobacter sp. and Agrobacterium sp. for DMP, DMI and DMT, respectively. This study illustrates that the detrimental DMPE and their natural metabolites may accumulate in the sulfate-reducing environment. Accordingly, proper surveillance program should be devised to monitor both the parent compounds and degradation intermediates of DMPE in order to protect the aquatic ecosystem and human health. PMID- 17462711 TI - Study of pyrene biodegradation capacity in two types of solid media. AB - Removal of pyrene, a representative PAH, was studied using laboratory tests in two different types of solid media: an organic matter collected on the surface of a vertical flow constructed wetland (VFCW) and a formulated clay silicate sand (inorganic matter). The aim of this study was to evaluate the capacity of pyrene biodegradation in these media in order to use them for treating run-off water. The sorption process, the kinetics of pyrene biodegradation and the influence of selected bacteria were also investigated. The sorption process was evaluated by adsorption isotherms and desorption kinetics using a batch equilibration method. The adsorption coefficient values of 28.8 and 2.1 for the organic and the inorganic matter respectively, confirmed the relationship of adsorption with organic carbon content. A small proportion of the sorbed pyrene was available for desorption (8% and 15% for the organic and the inorganic matter, respectively), indicating that sorption was partially irreversible, with the presence of hysteresis. For the formulated clay silicate sand inoculated with a specific bacteria (Mycobacterium sp.6PY1), selected for its ability to degrade PAHs, pyrene removal was complete in 32 days. With the organic matter, these values ranged from 40% to 95% for the different experiments, following a lag time of 3 weeks before observation of a significant degradation. Indigenous bacterial species in the organic medium had the metabolic capacity to degrade pyrene, and microbial populations pre-exposed to the PAH degraded pyrene faster than similar unexposed populations. Three metabolites of pyrene degradation by Mycobacterium were found. They accumulated in both organic and inorganic matter, indicating that the enzymes catalyzing them have slow kinetics. PMID- 17462712 TI - Application of real-time quantitative PCR for the detection of selected bacterial pathogens during municipal wastewater treatment. AB - Bacteria were detected at five stages of municipal wastewater treatment using TaqMan(R) real-time quantitative PCR (qPCR). Thirteen probe and primer sets were tested for diverse pathogens that may be present in wastewater, including Aeromonas hydrophila, Bacillus cereus, Clostridium perfringens, Enterococcus faecalis, Escherichia coli, E. coli O157:H7, Helicobacter pylori, Klebsiella pneumoniae, Legionella pneumophila, Listeria monocytogenes, Pseudomonas aeruginosa, Salmonella sp., and Staphylococcus aureus. The sensitivity of the assay was 100 fg of genomic DNA (=22 gene copies), based on a standard curve generated using A. hydrophila purified DNA. Samples from five stages of wastewater treatment were collected, including raw wastewater, primary effluents, mixed liquor, waste activated sludge and final effluents. In duplicate samples, E. coli, K. pneumoniae, C. perfringens and E. faecalis were detected throughout the wastewater process, and their numbers decreased by 3.52-3.98, 4.23-4.33, 3.15 3.39, and 3.24 orders of magnitude respectively, between the raw wastewater and final effluent stage. This qPCR method was effective for the detection of pathogens in wastewater and confirmed that the risk of exposure to pathogens in the wastewater discharge was well within the Environment Canada guidelines. PMID- 17462713 TI - Estimating sectoral pollution load in Lagos by Industrial Pollution Projection System (IPPS). AB - Sensitivity to environmental issues brought about increasing pressure from local community, groups, environmental organizations and government regulators on industries to reduce their pollutant emissions. In this study, Industrial Pollution Projection System (IPPS), which was developed by the Infrastructure and Environment Team of the World Bank, was used to estimate pollution load in ton/yr (with respect to employment) of industrial sectors in Lagos. The IPPS was developed to exploit the fact that the scale of industrial activity, its sectorial composition, and the process technologies, employed in production, heavily affect industrial pollution. Available data, from Manufacturer's Association of Nigeria (M.A.N.) for the years 1997-2002 was used for the estimation. From the cumulative ranking of the pollution load (ton/yr) estimate to all media (i.e. air, land, and water), Chemical and Pharmaceutical (CPH) sector is the highest polluting sector, followed by Basic Metal (BML), Domestic and Industrial Plastics (DIP), and Food, Beverage and Tobacco (FBT) sectors. Some of these sectors have the highest number of employees, and also appeared as the most polluting sectors in Lagos. PMID- 17462714 TI - The risk of recurrent cardiovascular events in patients with increased plasma homocysteine levels is reduced by short but not long-term therapy with folate and B vitamins. PMID- 17462715 TI - Electric injury model of murine arterial thrombosis. AB - BACKGROUND AND PURPOSE: Murine models of arterial thrombosis have gained utility with applications in genetically manipulated mice. Implementation of current models requires specialized equipment and provides limited outcome measures. A new murine model of continuously monitored arterial thrombosis was created. METHODS: An electric injury was delivered to the exterior surface of the common carotid artery using the flat end of a 140-mum steel microsurgery needle connected to the anode of a 3-V battery source. Direct current was delivered for 30 s. The developing thrombus was apparent as a white, platelet-dominated region at the site of injury. This region was continuously monitored and recorded by videotape for 30 min. Subsequently, the thrombus area was measured directly on the TV monitor, generating a time course for thrombogenesis. In a further evaluation of the model, three pharmacologically treated groups of mice were evaluated, with drug infusion immediately before thrombus induction: (1) saline (control), (2) heparin (60 units/kg), and (3) GR144053, a GPIIb/IIIa-specific antagonist (10 mg/kg). RESULTS: The basic model showed consistent thrombus development by 7-9 min, occasionally forming an occlusive thrombus. Most of the thrombi underwent one or more cycles of embolization and thrombus regrowth. In the experimental series, the heparin-treated group had a significantly decreased thrombus area versus controls (p<0.0001); the GR144053-treated mice had no apparent thrombus, supporting a dominant role of platelet aggregation in arterial thrombogenesis. CONCLUSION: This new model is simple to do, uses readily available instrumentation, and provides a continuously recorded quantifiable measure of thrombogenesis. PMID- 17462716 TI - Antiplatelet therapy for primary and secondary prevention in Jordanian patients with diabetes mellitus. AB - INTRODUCTION: Guidelines for diabetes treatment recommend the routine use of aspirin in patients with diabetes mellitus (DM) who have cardiovascular disease (CVD), those with at least one CVD risk factor, and for anyone with DM who is>30 years of age or, if contraindicated, an alternative antiplatelet agent. We evaluated the adherence to these recommendations in a subset of diabetic patients in Jordan, and identified physician and patient characteristics associated with the use of antiplatelet therapy. MATERIALS AND METHODS: A cross-sectional observational study conducted in randomly selected outpatient clinics. The study included 199 adult patients with DM who were eligible for antiplatelet prophylaxis. Review of medical charts and direct interviews with patients were performed to obtain information on demographic characteristics of the patients, clinical factors and medication prescription. Bivariate analysis and multivariate logistic regression methods were used to determine what factors were related to antiplatelet drug use. RESULTS: Among diabetic patients eligible for antiplatelet prophylaxis, the prevalence of antiplatelet drug use for primary or secondary prevention was 84%. Factors associated with antiplatelet prophylaxis were male gender (odds ratio, OR 2.97), presence of angina (OR 3.00), history of myocardial infarction (OR 3.75), history of coronary revascularization (OR 3.12), five or more prescription medications (OR 4.61), and physician's specialty other than endocrinology ( OR 4.35). CONCLUSIONS: Adherence rates to the international guidelines regarding routine antiplatelet prophylaxis in adults with DM who are eligible for such prophylaxis are suboptimal. Efforts towards increasing these rates of antiplatelet prophylaxis are necessary. PMID- 17462717 TI - Exploration of P2X3 in the rat stellate ganglia after myocardial ischemia. AB - ATP is implicated in peripheral pain signaling by actions on P2X receptors, especially P2X(3) receptor. Cardiac primary afferents running in the sympathetic nerves are considered to be essential pathways for transmission of cardiac nociception to the central nervous system. Because little is known about P2X(3) involvement in cardiac nociception, this study observed the difference in P2X(3) localization and expression in stellate ganglia (SG) from naive rats and in a pathological model of myocardial ischemic injury induced by repeated subcutaneous isoprenaline injections. Distribution of P2X(3) and morphometry of neurons in SG were investigated by immunohistochemistry, Western blotting, in situ hybridization (ISH) and by sterological study. Diffuse cytoplasmic P2X(3) immunolabelling was observed by light microsocopy. No nuclear labeling was detected. The intensity of P2X(3) labeling in the experimental myocardial ischemic injury group was increased in relation to that of the control group. Numerical densities of stellate ganglion neurons in the experimental group were higher than those of the control group. By Western blotting and ISH, the signals of P2X(3) protein and its mRNA in the myocardial ischemic group were higher than those of the control group. The P2X(3) labeling intensity and the numerical density in SG of the experimental myocardial ischemic injury group were enhanced, suggesting the involvement of P2X(3) receptor for the transmission of pain after myocardial ischemic injury. PMID- 17462718 TI - Pregnancy and dexamethasone: effects on morphometric parameters of gonadotropic cells in rats. AB - The effects of pregnancy and multiple dexamethasone (Dx) treatment on morphometric parameters of adenohypophyseal gonadotropic cells that produce follicle stimulating hormone (FSH) and luteinizing hormone (LH) were investigated in female Wistar rats. The rats in the experimental group received injections of 1.0, 0.5 and 0.5mg Dx/kg b.w. on days 16-18 of pregnancy, while the control group received equal volumes of saline. There was also an age-matched adult virgin control group. The experimental and control animals were sacrificed 24 and 72h after the last injection. Using the peroxidase-anti-peroxidase immunohistochemical labeling procedure, morphometric analyses showed that cell volume and volume density of FSH and LH cells on day 19 of pregnancy, as well as the number of LH cells, were significantly decreased compared to the virgin control values. On day 21 of gestation, the volume of FSH and LH cells remained smaller than in the virgin controls. Moreover, FSH and LH cell volume was significantly decreased 24h after multiple Dx treatment in comparison with the pregnant controls. Thus, during the last days of pregnancy, the morphometric parameters of gonadotropic cells decreased in comparison with the control virgin rats, but Dx treatment of pregnant rats had an inhibitory influence on FSH and LH cell size only 24h after the last dose. PMID- 17462719 TI - The changed immunolocalization of START-domain-containing 6 (StarD6) during the development of testes in rat perinatal hypothyroidism. AB - Thyroid hormones have an essential role in maintaining the normal developmental structure of testes during the neonatal stage. START-domain-containing 6 (StarD6) is exclusively expressed in germ cells during spermatogenesis; however, its biological role in rat perinatal hypothyroidism is not clear. After hypothyroidism was induced by daily administration of 0.05% 6-propyl-2-thiouracil (PTU), the pattern of StarD6 immunolocalization was examined from gestation day 15 to postnatal day 49. In normal rats, the labelling of StarD6 was confined to the germ cells from the third-week postpartum. In contrast, its immunoreactivity in hypothyroidal rats was not detected until the fourth-week postpartum. The immunolocalization pattern of StarD6 differed from that of normal adult rats during the seventh-week postpartum. StarD6 was clearly detected in the Leydig cells of the perinatal hypothyroid rats from the fifth-week postpartum. Therefore, StarD6 may play a pivotal role, not only in the spermatogenesis of normal rats, but also in the steroidogenesis of Leydig cells under perinatal hypothyroidism. PMID- 17462720 TI - An overview of laser microdissection technologies. AB - The development of laser-based tissue microdissection systems has provided the basis for the rapid acquisition of specific morphologically and/or phenotypically distinct types of cells for many types of molecular analysis. Two laser microdissection technologies based on distinct principles have been developed, namely: laser capture microdissection and laser cutting microdissection. This commentary will outline the principles of each system and indicate their main advantages and potential drawbacks. Also discussed will be methods of cell and tissue preparation with particular reference to fixation and staining, which are crucial to both successful laser-based microdissection and also downstream molecular studies. Laser microdissection techniques are powerful technologies which combine morphology and histochemistry with sophisticated molecular analysis. Through their appropriate application they have provided significant new insights into cell biology and pathology. PMID- 17462721 TI - The role of connexins in controlling cell growth and gene expression. AB - The purpose of this paper is to provide a brief overview of current thinking on the role of connexins, in particular Cx43, in growth regulation, and a more detailed discussion as to potential mechanisms involved with an emphasis on gene expression. While the precise molecular mechanism by which connexins can affect the growth of normal or tumor cells remains elusive, a number of exciting reports have expanded our understanding and are presented in some detail. Thus, we will discuss (Section 2): the role of protein-protein interactions in integrating connexins into multiple signal transduction pathways; phosphorylation at specific sites and reversal of growth inhibition; the role of the carboxy-terminal regulatory domain as a signaling molecule. Some of our latest work on the potential functions of endogenously produced carboxy-terminal fragments of Cx43 are also presented (Section 3). Finally, Section 4 will pay tribute to the rapidly emerging realization that connexins such as Cx43 and Cx32 exert important and extensive effects on gene expression, particularly those genes linked to growth regulation. PMID- 17462722 TI - The modulatory effects of connexin 43 on cell death/survival beyond cell coupling. AB - Connexins form a diverse and ubiquitous family of integral membrane proteins. Characteristically, connexins are assembled into intercellular channels that aggregate into discrete cell-cell contact areas termed gap junctions (GJ), allowing intercellular chemical communication, and are essential for propagation of electrical impulses in excitable tissues, including, prominently, myocardium, where connexin 43 (Cx43) is the most important isoform. Previous studies have shown that GJ-mediated communication has an important role in the cellular response to stress or ischemia. However, recent evidence suggests that connexins, and in particular Cx43, may have additional effects that may be important in cell death and survival by mechanisms independent of cell to cell communication. Connexin hemichannels, located at the plasma membrane, may be important in paracrine signaling that could influence intracellular calcium and cell survival by releasing intracellular mediators as ATP, NAD(+), or glutamate. In addition, recent studies have shown the presence of connexins in cell structures other than the plasma membrane, including the cell nucleus, where it has been suggested that Cx43 influences cell growth and differentiation. In addition, translocation of Cx43 to mitochondria appears to be important for certain forms of cardioprotection. These findings open a new field of research of previously unsuspected roles of Cx43 intracellular signaling. PMID- 17462723 TI - Recurrent gestational trophoblastic disease after hCG normalization following hydatidiform mole in The Netherlands. AB - OBJECTIVES: To determine the risk for recurrent trophoblastic disease after spontaneous normalization of human chorionic gonadotropin (hCG) levels in patients with hydatidiform mole and to determine the risk for tumor relapse after apparent remission following chemotherapy in patients with low- and high-risk persistent trophoblastic disease. METHODS: From 1994 until 2004, 355 patients with hydatidiform mole were registered at the Dutch Central Registry of Hydatidiform Mole and were monitored by sequential hCG assays in serum at the department of Chemical Endocrinology of the Radboud University Nijmegen Medical Centre. HCG regression curves were analyzed together with clinical information collected from the Hydatidiform Mole Database. RESULTS: Among the 355 registered hydatidiform mole patients, 265 patients attained spontaneous normalization following evacuation. Of the 265 patients, one patient (0.38%) subsequently required chemotherapeutic treatment for recurrent trophoblastic disease (95% confidence interval 0.0% to 2.1%). HCG levels did not decline to normal (<2.0 ng/ml) spontaneously in 90 patients; those patients were subsequently treated. Relapse rates were 8.1% (6/74) and 6.3% (1/16) for the low- and high-risk category respectively. CONCLUSION: Our analysis indicates that relapse risk in hydatidiform mole patients with spontaneous normalization is extremely low (one in 265 patients) after two normal hCG levels (<2.0 ng/ml) are achieved. Our results support the suggestion that two subsequent normal hCG levels may be sufficient to ensure sustained remission after hydatidiform mole evacuation. In contrary, in order to assure sustained remission, the relapse rates after chemotherapy in the current study emphasize the need for surveillance of trophoblastic tumor patients even after complete remission has apparently been achieved. PMID- 17462725 TI - Risk assessment of transmission of bovine viral diarrhea virus (BVDV) in abattoir derived in vitro produced embryos. AB - Bovine virus diarrhea virus (BVDV) is a pathogen of the bovine reproductive system causing reduced conception rates, abortions and persistently infected calves. Most if not all strains of BVDV are transmissible by natural mating and AI. For international trade, it is recommended that in vitro fertilized embryos be washed according to the IETS Manual. However, BVDV may not be entirely washed out, resulting in possible transmission risks to recipients. Donor cows, donor bulls and biological agents are all possible sources of contamination. The process for producing in vitro produced (IVP) embryos is complex and non standard, and some procedures can contribute to spread of BVDV to uninfected embryos. The structure of the zone pellucida (ZP) of IVP embryos permits adherence of BVDV to the ZP. To estimate the risk of producing infected recipients and persistently infected calves from abattoir-derived IVP embryos, a quantitative risk assessment model using Microsoft Excel and Palisade @Risk was developed. Assumptions simplified some of the complexities of the IVP process. Uncertainties due to incomplete or variable data were addressed by incorporating probability distributions in the model. Model variables included: disease prevalence; the number of donor cows slaughtered for ovaries; the number of oocytes collected, selected and cultured; the BVDV status of ovaries, semen, biological compounds and its behavior in the IVP embryo process. The model used the Monte Carlo method to simulate the IVP process. When co-culture cells derived from donor cows of unknown health status were used for in vitro culture (IVC), the probability of a recipient cow at risk of infection to BVDV per oocyte selected for IVP processing averaged 0.0006. However, when co-culture free from BVDV was used, the probability was 1.2 x 10(-5). Thus, for safe international trade in bovine IVP embryos (i.e. negligible risks of transmission of BVDV), co culture cells, if used during IVC for producing IVP embryos, should be disease free. PMID- 17462724 TI - Circadian mutant Overtime reveals F-box protein FBXL3 regulation of cryptochrome and period gene expression. AB - Using a forward genetics ENU mutagenesis screen for recessive mutations that affect circadian rhythmicity in the mouse, we isolated a long period (approximately 26 hr) circadian mutant named Overtime (Ovtm). Positional cloning and genetic complementation reveal that Ovtm is encoded by the F-box protein FBXL3, a component of the SKP1-CUL1-F-box-protein (SCF) E3 ubiquitin ligase complex. The Ovtm mutation causes an isoleucine to threonine (I364T) substitution leading to a loss of function in FBXL3, which interacts specifically with the CRYPTOCHROME (CRY) proteins. In Ovtm mice, expression of the PERIOD proteins PER1 and PER2 is reduced; however, the CRY proteins CRY1 and CRY2 are unchanged. The loss of FBXL3 function leads to a stabilization of the CRY proteins, which in turn leads to a global transcriptional repression of the Per and Cry genes. Thus, Fbxl3(Ovtm) defines a molecular link between CRY turnover and CLOCK/BMAL1 dependent circadian transcription to modulate circadian period. PMID- 17462726 TI - Relative risks and approaches to biosecurity in the use of embryo technologies in livestock. AB - Embryo technologies have been integrated into production systems for a variety of livestock species. As relates to transmission of infectious diseases, our working hypothesis has been that use of embryo transfer for distribution of germ plasm within and between herds and flocks is likely safer than the movement of postnatal animals. Indeed, research and experience generally have been supportive of this hypothesis. However, the relative risks of transmitting infectious agents via embryo transfer vary among donor species. Further, different methods of producing embryos appear to present different risks. This paper provides a comparative overview of the risks of transmitting infectious diseases via transfer of both in vivo- and in vitro-derived embryos in common domesticated livestock species. Also discussed are universal approaches to biosecurity in embryo production and transfer. PMID- 17462727 TI - Adolescent psychological well-being by identity style. AB - High school-age adolescents (N=262) in two samples completed a measure of identity style, along with measures of self-esteem, hopelessness, delinquent attitudes, educational expectations, and optimism/efficacy. Analysis of variance was used to evaluate the relationship between identity style and the other psychosocial variables. Participants classified as having a diffuse/avoidant orientation had lower self-esteem and higher delinquent attitude scores than those with either an information or normative orientation. At the same time, the diffuse-avoidant style was associated with higher hopelessness and lower optimism/efficacy scores than the normative and information styles. The normative and information styles did not differ significantly in terms of the dependent variables. Results also indicated that males were more likely to be diffused and females more likely to be information-oriented. PMID- 17462728 TI - The performance of a bone-derived scaffold material in the repair of critical bone defects in a rhesus monkey model. AB - The efficacy and safety of a material derived from human bones in the repair of critical segmental bone defects are evaluated in a rhesus monkey model. Frozen human bones were chemically and physically processed into a partially demineralized and deproteinized material in blocks. The complete tissue engineered (TE) bone was constructed of the material preseeded with allogeneic bone marrow mesenchymal stem cells (MSCs). The material alone and the TE bone were, respectively, implanted to bridge 2.5cm-long critical defects in right and left radii of 15 monkeys. At weeks 1, 2, 3, 6 and 12 post-implantation, the grafts were collected from three animals and assessed for the local expression of osteogenic markers, histological and roentgenographic features, and immune reactions. It was shown that defects were well repaired with both treatments whereas the bone defects in 2 additional untreated animals remained the same size after 12 weeks. In radii implanted with the TE bones, the repair processes were approximately 3 weeks faster and new bones were formed in a multipoint way. There was neither observable toxic effect nor overt immune rejection in any animals. Taken together, these observations suggest that the TE bone blocks composited of the allogeneic or xenogeneic bone-derived scaffold and allogeneic MSCs may provide an ideal method for repairing large segmental bone defects. PMID- 17462729 TI - Effects of calcium ion incorporation on bone healing of Ti6Al4V alloy implants in rabbit tibiae. AB - The biocompatibility of calcium ion (Ca)-incorporated Ti6Al4V alloy implants, produced by hydrothermal treatment using a Ca-containing solution, was investigated. The surface characteristics were evaluated by scanning electron microscopy, thin-film X-ray diffractometry, Auger electron spectroscopy, and stylus profilometry. The viability of MC3T3-E1 cells on Ca-incorporated machined Ti6Al4V surfaces with different oxide thicknesses was compared with that on untreated machined Ti6Al4V surfaces with MTT assay. The osteoconductivity of the Ca-incorporated Ti6Al4V implants was evaluated by removal torque testing and histomorphometric analysis after 6 weeks of implantation in rabbit tibiae. Our results show that hydrothermal treatment with a Ca-containing solution produced a crystalline CaTiO(3) layer on Ti6Al4V surfaces, and calcium ions were gradually incorporated throughout the oxide layer. After immersion in Hank's balanced salt solution, a considerable apatite deposition was observed on all surfaces of the Ca-incorporated samples. Significant increases in cell viability (p<0.001), removal torque forces, and bone-to-implant contact values (p<0.05) were observed for Ca-incorporated Ti6Al4V implants compared with those for untreated Ti6Al4V implants. PMID- 17462730 TI - What is the best salvage therapy for treatment of isolated CNS relapse in elderly patients with imatinib-responsive Ph(+) ALL? AB - We report the case of an elderly patient affected by Philadelphia positive Acute Lymphoblastic Leukaemia (Ph(+) ALL) who developed meningeal leukaemia during imatinib monotherapy, despite bone marrow molecular remission. Aggressive central nervous system (CNS)-directed therapy in combination with continued imatinib treatment might be, at the moment, the most effective salvage therapy for imatinib-responsive elderly patients with isolated CNS relapse. In view of the inefficacy of imatinib at preventing meningeal leukaemia for its poor penetration into the CNS, CNS prophylactic therapy should always be an integral part of any imatinib-based treatment strategy for Ph(+) ALL. PMID- 17462731 TI - Guanine nucleotide depletion inhibits pre-ribosomal RNA synthesis and causes nucleolar disruption. AB - Inosine monophosphate dehydrogenase (IMPDH) is a pivotal enzyme in the de novo pathway of guanine nucleotide biosynthesis. Inhibitors of this enzyme decrease intracellular guanine nucleotide levels by 50-80% and have potential as anti neoplastic agents. Both mycophenolic acid (MPA) and AVN-944 are highly specific inhibitors of IMPDH that cause cell cycle arrest or apoptosis in lymphocytes and leukemic cell lines. We have examined the mechanisms by which these two agents cause cytotoxicity. Both MPA and AVN-944 inhibit the growth of K562 cells, and induce apoptosis in Raji B and CCRF-CEM T cells. Both compounds strikingly inhibit RNA synthesis within 2 h of exposure. Depletion of guanine nucleotides by MPA and AVN-944 also causes an early and near-complete reduction in levels of the 45S precursor rRNA synthesis and the concomitant translocation of nucleolar proteins including nucleolin, nucleophosmin, and nucleostemin from the nucleolus to the nucleoplasm. This efflux correlates temporally with the sustained induction of p53 in cell lines with wild-type p53. We conclude that inhibition of IMPDH causes a primary reduction in rRNA synthesis and secondary nucleolar disruption and efflux of nucleolar proteins that most likely mediate cell cycle arrest or apoptosis. The ability of AVN-944 to induce apoptosis in a number of leukemic cell lines supports its potential utility in the treatment of hematologic malignancies. PMID- 17462732 TI - Immunoglobulin light chain repertoire in hairy cell leukemia. AB - Of 166 hairy cell leukemia (HCL) patients, 81 had kappa and 80 had lambda expression. IGKV-J and IGLV-J rearrangement structure was analyzed in 21 HCL patients (11 kappa, 10 lambda). For kappa, IGKV1-5 was most frequent, and the KJ2 gene was over-utilized. For lambda HCL, LJ3 was over-utilized compared to normal. This study significantly adds to previous studies of light chain usage in HCL and is the first to report light chain gene usage. In HCL, we confirm the lack of kappa predominance observed in normal lymphocytes and in chronic lymphocytic leukemia, and note over-representation of several light chain genes. PMID- 17462733 TI - Canine cathelicidin (K9CATH): gene cloning, expression, and biochemical activity of a novel pro-myeloid antimicrobial peptide. AB - Cathelicidins, a group of cationic peptides found in leukocytes and epithelial cells, play a central role in the early innate immune defense against infection. Although these host defense peptides have been reported in several mammalian species, including primates, no cathelicidins have been identified in carnivores. Here we report the cloning, tissue expression and biological activity of a novel canine cathelicidin (K9CATH). The full-length cDNA sequence of K9CATH encodes a predicted 172 amino acid pre-propeptide that is 60-70% similar to other mammalian cathelicidins. Mass spectrometry analysis confirmed that the 38 aa mature K9CATH peptide was present in neutrophil granule contents. Synthetic K9CATH displayed broad antimicrobial activity against Gram-positive bacteria (Listeria monocytogenes, and Staphylococcus aureus; MICs (minimal inhibitory concentrations) 0.5 and 50 microM, respectively), Gram-negative bacteria (Escherichia coli, Klebsiella pneumoniae, Salmonella serotype Typhimurium, Pseudomonas aeruginosa, Proteus mirabilis; MICs 1.25 microM, Salmonella serotype Enteritidis; MIC 0.5 microM, and Neisseria gonorrhoeae; MIC 0.06 microM), and yeast (Candida albicans; MIC 12.5-50 microM). K9CATH demonstrated high antimicrobial activity against Ureaplasma canigenitalium, and lower activity against Ureaplasma urealyticum (MIC 0.06 and 50 microM, respectively). Similar to its ovine congener SMAP-29, K9CATH possesses salt-independent antimicrobial activity and LPS binding capacity. K9CATH displayed minimal hemolytic activity against human, dog and chicken erythrocytes. The potency and broad antimicrobial activity of K9CATH suggest that this peptide may act as a fundamental contributor to the innate immune responses in this carnivore species. PMID- 17462734 TI - The role of mood stabilisers in the treatment of the depressive facet of bipolar disorders. AB - It was previously shown that available mood stabilisers are used to treat bipolar depression. As part of the natural course of illness, patients with bipolar disorder often suffer from episodes of depression more frequently and for longer durations than mania. A major challenge in the treatment of bipolar depression is the tendency for antidepressant medications, particularly tricyclic antidepressants, to precipitate episodes of mania, or to increase cycle frequency or symptom intensity. Thus, exploring the utility of mood stabilisers as monotherapy for bipolar depression is important. The aim of this review it to collate data involving the effects of some mood stabilisers like lithium, carbamazepine, valproate and lamotrigine in depressive aspects of bipolar disorder, but as well using an animal model of depression, to understand their mechanism of action. PMID- 17462736 TI - Stable carbon isotope ratios of POPs--a tracer that can lead to the origins of pollution. AB - An elemental analyzer in combination with stable isotope ratio mass spectrometry (EA-IRMS) was used to demonstrate that two samples of the chloropesticide Melipax discovered in two countries varied only by 0.1 per thousand in their delta 13C values. Gas chromatography interfaced to a stable isotope mass spectrometer (GC IRMS) was used to study the delta 13C values of individual congeners in two technical polybrominated diphenyl ether products, i. e. DE-71 and DE-79, at three different concentration levels. Injections of 4 microg and 5.2 microg of DE-71 and DE-79 were most suited for the determinations. In both products, the delta 13C values were usually the more negative the higher the degree of bromination was. The delta 13C values of three hexabromo isomers BDE 138, BDE 153, and BDE 154 could be determined in both products. In all cases the delta 13C values of the three hexaBDEs were more depleted in delta 13C in the higher brominated technical octabromo diphenyl ether DE-79. PMID- 17462737 TI - Enthalpy-entropy compensation in the transition of a monospecific antibody towards antigen-binding promiscuity. AB - In a recent article published in Molecular Immunology (Dimitrov, J.D., Lacroix Desmazes, S., Kaveri, S.V., Vassilev, T.L., 2007. Transition towards antigen binding promiscuity of a monospecific antibody. Mol. Immunol. 44, 1854-1863.), the authors perform kinetic and thermodynamic analyses using a surface plasmon resonance-based technique to explain polyspecificity of antibodies in terms of molecular mechanisms. They found that after the antibody was exposed to urea, a protein destabilizing agent, it had the same affinity as the native state and that the two forms of the antibody use two distinct thermodynamic pathways for binding to the same antigen. In this letter, we report our analysis of these results in terms of equilibrium thermodynamics to show that the specificity transition of the antibody exhibits enthalpy-entropy compensation. PMID- 17462738 TI - Technical appendix: cost-effectiveness of vitamin therapy for age-related macular degeneration. PMID- 17462739 TI - Laser peripheral iridotomy in eyes with narrow drainage angles: ultrasound biomicroscopy outcomes. The Liwan Eye Study. AB - PURPOSE: To assess the short-term effect of laser peripheral iridotomy (LPI) on anterior segment anatomy in angle-closure suspects using ultrasound biomicroscopy (UBM). DESIGN: Prospective intervention study. PARTICIPANTS: Persons identified as angle-closure suspects aged 50-79 years from a population-based survey in Guangzhou, China. INTERVENTION: Laser peripheral iridotomy was performed on 1 randomly selected eye. Ultrasound biomicroscopy examination was carried out before and 2 weeks after the intervention. MAIN OUTCOME MEASURES: Proportion of eyes with iridotrabecular contact (ITC), as well as changes in UBM parameters including angle opening distance (AOD), iris thickness (IT), iris curvature, iris ciliary process distance, trabecular-ciliary process distance (TCPD), and scleral spur to iris insertion distance (SS-IR). RESULTS: A total of 72 of 101 eligible subjects participated in the study. The proportion of people with UBM-identified ITC in > or =1 quadrant dropped from 95% (68/72) before to 59% (42/72) after LPI. After LPI, the mean AOD at 250 microns increased from 0.064 mm (standard deviation [SD], 0.052) to 0.085 (0.052) mm (P<0.001); angle recess area increased from 0.040 (0.030) to 0.070 (0.036) mm2 (P<0.0001); TCPD increased from 0.537 to 0.561 mm (P = 0.001); IT at 750 microns increased from 0.440 to 0.459 mm (P = 0.094), and IT at 1000 microns increased from 0.471 to 0.488 mm (P = 0.0001). Eyes whose angles remained closed after LPI (pigmented trabecular meshwork not visible in > or =3 quadrants) tended to have shallower AOD both at 250 (0.071 vs. 0.049 mm; P = 0.09) and 500 microns (0.108 vs. 0.052 mm; P = 0.001), a thicker iris (IT at 750 microns, 0.447 vs. 0.415 mm; P = 0.041), a more anterior positioned ciliary body (TCPD, 0.514 vs. 0.562 mm; P = 0.03), and a statistically nonsignificant more anterior iris insertion (SS-IR: 0.085 vs. 0.125 mm; P = 0.061), before LPI. CONCLUSIONS: Laser peripheral iridotomy results in a significant increase in the angle width in Chinese people with narrow angles. However, some iridotrabecular contact was found in 59% of eyes with a patent iridotomy. This was associated with smaller anterior chamber angle dimensions and a thicker iris, both of which may play a causative role in maintaining angle closure after LPI. PMID- 17462740 TI - Cleavage of an RNA analog by Zn(II) macrocyclic catalysts appended with a methyl or an acridine group. AB - Two macrocycles (1 and 2) are prepared that incorporate pendent groups in macrocycle 3 (3=1-oxa-4,7,10-triazacyclododecane) with the goal of studying the effect of these pendent groups on metal ion complexation, solution chemistry and catalysis. Zn(1) contains a macrocyclic ligand with a pendent acridine group and Zn(2) has an appended methyl group. Water ligand pK(a) values for Zn(1) (6.7) and Zn(2) (7.3) are lower than that of Zn(3) (7.7). Zn(II) complexes of 1 and 2 are studied as catalysts for the cleavage of 2-hydroxypropyl 4-nitrophenylphosphate (HpPNP), an RNA analog. Zn(2) has a lower catalytic activity over the pH range 7 10 for cleavage of HpPNP compared to the parent macrocyclic complex, Zn(3). In contrast, Zn(1) has a threefold larger rate constant at pH 7.0 compared to Zn(2), attributed to the presence of a catalytic species which has a protonated acridine amino group. The binding constant of 1.5mM at pH 8.0 for formation of the Zn(2) uridine adduct is similar to that for Zn(3), suggesting that N-alkylation of the macrocyclic ligand does not interfere with binding of the Zn(II) complex to uridine groups. Binding of cytidine to Zn(2) was not detectable under similar conditions up to 25mM nucleoside. Binding experiments under similar conditions could not be carried out for adenosine or guanosine due to their low solubility. PMID- 17462741 TI - Physicochemical study of floranol, its copper(II) and iron(III) complexes, and their inhibitory effect on LDL oxidation. AB - The antioxidant activity of floranol (3,5,7,2'-tetrahydroxy-6-methoxy-8 prenylflavanone), a new flavonoid isolated from the roots of Dioclea grandiflora, was evaluated by the inhibition of human low-density lipoprotein (LDL) oxidation. Floranol increased its oxidation lag-phase significantly in a dose-dependent manner. As the antioxidant mechanism may involve metal coordination, we have undertaken a detailed study of floranol interactions with Cu(II) and Fe(III) by combination of UV-visible (UV-Vis) and mass spectrometries and cyclic voltammetry. The acidity constants of the ligand as well as the stability constants of the metal complexes were calculated. The pKa values of 6.58, 11.97 and 13.87 were determined and the following acidity order is proposed 7-OH>5 OH>2'-OH. The best fit between experimental and calculated spectra was obtained assuming the formation of two Cu(II) complexes: [CuL] logbeta=19.34+/-0.05 and [CuL(2)](2-) logbeta=26.4+/-0.10 and three Fe(III) complexes: [FeL(3)](3-) logbeta=44.72+/-0.09, [FeL(2)](-) logbeta=35.32+/-0.08 and [FeL](+) logbeta=19.51+/-0.04. In addition, copper and iron reduction is less favorable in the presence of floranol. These results indicate that floranol can efficiently bind Cu(II) and Fe(III) ions thus preventing their effect on LDL oxidation. PMID- 17462742 TI - Golgi's contribution to medicine. AB - The historical analysis of Golgi's research work reveals that his contribution to the progress of science is not confined to neurosciences and to cellular biology. In fact, Golgi was a passionate and a skillful student of medical problems, in particular of those posed by infectious diseases. Golgi approached these problems with a rigorous method of observation and experimentation with the aim of understanding the genesis of the symptoms and their significance for the progress of the disease. His fundamental contribution to medicine is to be found in his researches on malaria, in which he clarified the biological cycle of the parasite within the blood cells, and discovered the relation between the biological cycle of the parasite and the onset of the fever, thus providing the basis for therapy. PMID- 17462743 TI - Dissociation of emotional decision-making from cognitive decision-making in chronic schizophrenia. AB - Recent studies have examined the decision-making ability of schizophrenic patients using the Iowa Gambling Task (IGT). These studies, however, were restricted to the assessment of emotional decision-making. Decision-making depends on cognitive functions as well as on emotion. The purpose of this study was to examine the performance of schizophrenic patients on the IGT and the Game of Dice Task (GDT), a decision-making task with explicit rules for gains and losses. In addition, it was intended to test whether poor performance on IGT is attributable to impairments in reversal learning within the schizophrenia group using the Simple Reversal Learning Task (SRLT), which is sensitive to measure the deficit of reversal learning following ventromedial prefrontal cortex damage. A group of 23 stable schizophrenic patients and 28 control subjects performed computerized versions of the IGT, GDT, SRLT and Wisconsin Card Sorting Test (WCST). While schizophrenic patients performed poorly on the IGT relative to normal controls, there was no significant difference between the two groups on GDT performance. The performance of the schizophrenia group on the SRLT was poorer than that of controls, but was not related to IGT performance. These data suggest that schizophrenic patients have impaired emotional decision-making but intact cognitive decision-making, suggesting that these two processes of decision making are different. Furthermore, the impairments in reversal learning did not contribute to poor performance on the IGT in schizophrenia. Therefore, schizophrenic patients have difficulty in making decisions under ambiguous and uncertain situations whereas they make choices easily in clear and unequivocal ones. The emotional decision-making deficits in schizophrenia might be attributable more to another mechanism such as a somatic marker hypothesis than to an impairment in reversal learning. PMID- 17462744 TI - Dissociative disorders among alcohol-dependent inpatients. AB - The aim of this study was to determine the prevalence of dissociative disorders among inpatients with alcohol dependency. The Dissociative Experiences Scale was used to screen 111 alcohol-dependent patients consecutively admitted to the inpatient unit of a dependency treatment center. Subgroups of 29 patients who scored 30.0 or above and 25 patients who scored below 10.0 were then evaluated with the Dissociative Disorders Interview Schedule and the Structured Interview for DSM-IV Dissociative Disorders. The interviewers were blind to the Dissociative Experiences Scale scores. Of the 54 patients evaluated, 10 (9.0% of the original 111) patients had a dissociative disorder. A considerable number of the remaining patients reported a high level of dissociative experiences. Among the dissociative disorder group, nine patients had dissociative disorder not otherwise specified and one patient had depersonalization disorder. Female gender, younger age, history of suicide attempt, childhood emotional and sexual abuse, and neglect were more frequent in the dissociative disorder group than among non-dissociative patients. The dissociative disorder group also had somatization disorder, borderline personality disorder, and lifetime major depression more frequently. For 9 of the 10 dissociative patients, dissociative symptoms started before the onset of alcohol use. Although the probability of having a comorbid dissociative disorder was not higher among alcohol-dependent inpatients than among the general psychiatric inpatients, the dissociative subgroup had distinct features. Many patients without a dissociative disorder diagnosis (predominantly men) provided hints of subtle dissociative psychopathology. Implications of comorbid dissociative disorders and dissociative experiences on prevention and treatment of alcohol dependency and the importance of gender-specific characteristics in this relationship require further study. PMID- 17462745 TI - The proteasome function is required for Mycobacterium leprae-induced apoptosis and cytokine secretion. AB - Previous studies have demonstrated the importance of the ubiquitin-proteasome pathway in the immune response to bacterial pathogens. To investigate the role of this system in the context of leprosy, Mycobacterium leprae-stimulated peripheral blood mononuclear cells (PBMC) were treated with the proteasome inhibitor MG132 to assess the levels of apoptosis and cytokine secretion. The results showed that the inhibition of proteasome activity significantly reduced M. leprae-mediated cell death. In addition, MG132 treatment led to a significant decrease in M. leprae-induced TNF-alpha and IL-10 secretion. Together, these results suggest that modulations of the ubiquitin-proteasome pathway may participate in the human response to M. leprae. PMID- 17462746 TI - Adenylate kinase 5 autoimmunity in treatment refractory limbic encephalitis. AB - We report two men with limbic encephalitis (LE) refractory to corticosteroids, IVIg and plasma exchange. Both patients had serum/CSF antibodies that reacted with the cytoplasm of neurons. Probing of a hippocampal cDNA library resulted in the isolation of adenylate kinase 5 (AK5). Patients' antibodies, but not those of 111 controls, recognized AK5-expressing phage plaques. Human AK5-affinity purified antibodies reproduced the neuronal immunolabeling of patients' antibodies, and co-localized with a rabbit AK5 antibody, confirming that the brain autoantigen was AK5. Detection of antibodies to AK5 in LE patients carries a poor prognosis, and suggests the prompt use of aggressive immunosuppression. PMID- 17462747 TI - Genotyping of hepatitis C virus isolates by a new line probe assay using sequence information from both the 5'untranslated and the core regions. AB - The correct assessment of hepatitis C virus (HCV) genotypes and subtypes by commercial assays is of utmost importance mainly for the therapeutic management of patients suffering from HCV infections. In this study, the performance characteristics of a newly designed genotyping assay were evaluated that does not rely exclusively on sequence information derived from the 5'untranslated region but also takes into account part of the HCV core. One hundred and ten clinical specimens were tested by this new assay prior to its commercialisation. The obtained typing results were compared to those recorded by the 5'UTR-based Versant HCV Genotyping Assay, version 1, the core-related Gen-Eti K DEIA, and phylogenetic analyses of partial HCV core and NS5B sequences. The HCV genotypes and subtypes identified by the newly devised kit were completely in line with the assignments achieved by DEIA and phylogenetic analyses. In particular, all 64 HCV strains belonging to subtypes 1a or 1b were recognised correctly, and HCV 6e and 6f isolates were adequately assigned to subtypes 6c-l. Thus, the second generation of the Versant genotyping assay could overcome the drawbacks of its exclusively 5'UTR-based predecessor and will turn out to be a reliable tool for HCV typing in clinical laboratories. PMID- 17462748 TI - The topographic brain: from neural connectivity to cognition. AB - A hallmark feature of vertebrate brain organization is ordered topography, wherein sets of neuronal connections preserve the relative organization of cells between two regions. Although topography is often found in projections from peripheral sense organs to the brain, it also seems to participate in the anatomical and functional organization of higher brain centers, for reasons that are poorly understood. We propose that a key function of topography might be to provide computational underpinnings for precise one-to-one correspondences between abstract cognitive representations. This perspective offers a novel conceptualization of how the brain approaches difficult problems, such as reasoning and analogy making, and suggests that a broader understanding of topographic maps could be pivotal in fostering strong links between genetics, neurophysiology and cognition. PMID- 17462749 TI - The effect of luminal ghrelin on pancreatic enzyme secretion in the rat. AB - Ghrelin, a 28-amino-acid peptide produced predominantly by oxyntic mucosa has been reported to affect the pancreatic exocrine function but the mechanism of its secretory action is not clear. The effects of intraduodenal (i.d.) infusion of ghrelin on pancreatic amylase outputs under basal conditions and following the stimulation of pancreatic secretion with diversion of pancreato-biliary juice (DPBJ) as well as the role of vagal nerve, sensory fibers and CCK in this process were determined. Ghrelin given into the duodenum of healthy rats at doses of 1.0 or 10.0 microg/kg increased pancreatic amylase outputs under basal conditions or following the stimulation of pancreatic secretion with DPBJ. Bilateral vagotomy as well as capsaicin deactivation of sensory fibers completely abolished all stimulatory effects of luminal ghrelin on pancreatic exocrine function. Pretreatment with lorglumide, a CCK(1) receptor blocker, reversed the stimulation of amylase release produced by intraduodenal application of ghrelin. Intraduodenal ghrelin at doses of 1.0 or 10.0 microg/kg increased plasma concentrations of CCK and ghrelin. In conclusion, ghrelin given into the duodenum stimulates pancreatic enzyme secretion. Activation of vagal reflexes and CCK release as well as central mechanisms could be implicated in the stimulatory effect of luminal ghrelin on the pancreatic exocrine functions. PMID- 17462750 TI - Characterization of the Drosophila insulin receptor promoter. AB - The insulin receptor (InR) signaling pathway is largely conserved in metazoans and it is required for normal growth and development in Drosophila. Despite the importance of this pathway in regulating growth, development and metabolism in Drosophila, little is known about how dInR expression is controlled in flies. Here we report the characterization of the dInR gene promoter and the analysis of its expression during embryo development. Drosophila InR gene has three promoters spanning 40 kb in the genome. These promoters direct the expression of three distinct mRNA transcripts that share common exons downstream of the initiator codon ATG but have different 5'UTRs. All three promoters are differentially regulated, spatially and temporally, contributing to a very complex pattern of expression in the developing embryo. Our results indicate that dInR expression in Drosophila displays an intricate pattern of regulation that assures an adequate control of growth, development and metabolism. PMID- 17462751 TI - Secondary hypertension: current diagnosis and treatment. AB - Secondary hypertension affects a small but significant number of the hypertensive population and, unlike primary hypertension, is a potentially curable condition. The determinant for workup is dependent on the index of suspicion elicited during patient examination and treatment. Specific testing is available and must be balanced depending on the risk and cost of the workup and treatment with the benefits obtained if the secondary cause is eliminated. This article reviews common manifestations, workup, and the current treatments of the common causes of secondary hypertension. PMID- 17462752 TI - Stroke after surgical treatment of sinus venosus type atrial septal defect: percutaneous treatment. AB - BACKGROUND AND PURPOSE: Sinus venosus-type atrial septal defect can be associated with anomalous drainage of the upper right pulmonary vein into the superior vena cava. Surgical correction provides reconstruction of superior vena cava and rerouting of the pulmonary vein into the left atrium. SUMMARY OF CASE: We report the case of a young woman who experienced 2 strokes 11 years after surgical repair of sinus venosus-type atrial septal defect. CONCLUSIONS: Interventional treatment resolved the superior vena cava stenosis, rerouted the pulmonary vein into the left atrium and abolished the right to left shunt. PMID- 17462753 TI - Effect of statins on contrast-induced nephropathy in patients with acute myocardial infarction treated with primary angioplasty. AB - We investigated whether pretreatment with statin may prevent contrast-induced nephropathy in patients who underwent primary coronary intervention for acute myocardial infarction (AMI). A total of 279 consecutive patients who underwent successful primary angioplasty for a first AMI were studied. Contrast-induced nephropathy was defined as an increase in serum creatinine of > or =5 mg/dL after the primary PCI. 56 patients receiving statin treatment before admission had lower incidence of the contrast-induced nephropathy than those without it (7.1% and 20.6%, P<0.01). Multivariable logistic regression analysis revealed that absence of statin pre-treatment was a significant predictor of the reperfusion arrhythmia along with anterior AMI, baseline creatinine value, time-to reperfusion, higher volume of the contrast agent. Pre-treatment with statin could reduce the contrast-induced nephropathy after primary coronary intervention in patients with AMI. PMID- 17462754 TI - Pulmonary artery dissection and conservative medical management. AB - Pulmonary artery dissection is a rare clinical entity, which has been related to pulmonary arterial hypertension. It is frequently presented as cardiogenic shock or sudden death, so diagnosis is often made at autopsy. The management with best results is surgery. We report a case of pulmonary artery dissection associated with previous aortic valve replacement with a favourable outcome, using conservative medical therapy. PMID- 17462755 TI - Snake venom for acute myocardial infarction: natural serendipity or an obstruction to treatment? AB - A 55-year-old man developed an acute myocardial infarction and was bitten by a snake with haemotoxic venom prior to any treatment being given. We discuss our treatment of myocardial infarction in a patient who had been unintentionally anticoagulated. PMID- 17462756 TI - High-frequency QRS electrocardiogram analysis during exercise stress testing for detecting ischemia. AB - INTRODUCTION: ECG stress testing is an inexpensive and non-invasive detector of myocardial ischemia; addition of high-frequency QRS analysis (HFQRS) may improve accuracy. This study compared HFQRS during exercise in patients with and without ischemia as defined by multiple criteria. MATERIAL AND METHODS: High-resolution ECGs were recorded for 139 patients undergoing T99-sestamibi/T201-thallium stress testing. Twenty-three were positive by at least two and 37 were negative for ischemia by all three of the following criteria: nuclear scan, ST-segment analysis and typical angina. Sixty-four not meeting criteria for positive or negative, six with adenosine test and nine patients with ECG recording artifacts were excluded. Mean age of the study group was 62+/-10 years, 83% were male. Ischemic patients had a higher incidence of previous myocardial infarction and coronary intervention than non-ischemic patients (74% vs. 46%; P=0.03 and 70% vs. 43%; P=0.05, respectively), but had a lower body mass index (28.7+/-5 vs. 33.0+/ 8; P=0.015). HFQRS analysis consisting of signal averaging (150-250 Hz) and calculation of root mean squared values for each lead at different time points was performed and was similar between the groups. The relative change in HFQRS (RCQ) was calculated for each lead: {(maxHFQRS-minHFQRS)/maxHFQRS}. For each patient an RCQ index was calculated by averaging the two leads with the greatest RCQ value. The RCQ index was greater in ischemic vs. non-ischemic patients (45% vs. 34%; P=0.0069). CONCLUSION: Maximum decrease in HFQRS, as quantified by RCQ index, was greater in ischemic vs. non-ischemic patients. Use of the RCQ index may improve the diagnosis of ischemia during exercise stress testing. PMID- 17462757 TI - Obesity can disqualify would-be parents from adoption--a novel reason for not getting fat. PMID- 17462758 TI - Documentation of early improvement of left ventricular function in tako-tsubo cardiomyopathy. PMID- 17462759 TI - Unique wall motion abnormalities on stress echocardiogram associated with Wolff Parkinson-White pattern electrocardiogram: a case report. AB - Preexcitation syndromes pose a diagnostic challenge for exercise stress testing. We present stress echocardiographic findings in patient with Wolff-Parkinson White (WPW) pattern on surface electrocardiogram (EKG). Echocardiogram at rest demonstrated paradoxical septal motion which disappeared during peak exercise and reappeared during rest. Preexcitation pattern on EKG also disappeared at peak stress. We tried to explain the underlying mechanisms of the above abnormality and the usefulness of stress testing in this group of patients. PMID- 17462760 TI - Risk factors for prolonged intensive care treatment following atrial septal defect closure in adults. AB - BACKGROUND: Today, percutaneous or surgical closure of atrial septal defects (ASD) in adults are considered effective and safe treatments. However, some cases of severe left ventricular dysfunction after ASD closure were observed. This study aims at identifying predictors for prolonged intensive care unit stay, and postoperative inotropic support after ASD closure. METHODS: Records of 281 adult patients who had undergone surgical closure of a secundum ASD between 1974 and 2000 at an age over 30 years (mean 43.8, maximum 76 years) were reviewed retrospectively. The endpoints were defined as prolonged intensive care unit stay (>2 days), and postoperative inotropic support (Dopamine, Dobutamine or Adrenalin). RESULTS: Thirty-day mortality rate was 0.7% (2 patients). Prolonged intensive care unit stay was observed in 70 patients (25%). Postoperative inotropic support was necessary in 84 patients (30%). Independent risk factors for prolonged intensive care unit stay in multivariate analysis were preoperative atrial fibrillation (p=0.011), and larger ASD (p=0.026). Older age at operation (p<0.001) and longer time on extracorporeal circulation (p<0.001) emerged as independent risk factor for postoperative use of inotropic support in multivariate analysis. CONCLUSIONS: Surgical ASD closure in adults is usually safe. However, a distinct subgroup of patients is at risk for prolonged intensive care treatment. Timely closure of the ASD must be advised since older age emerged as a predictor for postoperative use of inotropic support. Since atrial fibrillation is a strong independent risk factor for prolonged intensive care unit stay the preservation of sinus rhythm must be aimed at. PMID- 17462761 TI - An epsilon wave in arrhythmogenic right ventricular cardiomyopathy/dysplasia. AB - A 53-year-old male presented to our emergency department with a sudden onset of ventricular tachycardia with left bundle branch block and right axis deviation. After the tachyarrhythmia converting to sinus rhythm, the ECG displayed sinus rhythm with a typical epsilon wave in leads V1 and V2. Arrhythmogenic right ventricular dysplasia/cardiomyopathy (ARVD/C) was suspected. The epsilon wave is the most specific hallmark for the diagnosis of ARVD/C, although it is insensitive. For the management of these patients, antiarrhythmic medications appear to be effective. However, implantable cardioverter defibrillators are the only reliable treatment to prevent sudden cardiac death. PMID- 17462762 TI - Premature myocardial infarction: clinical profile and angiographic findings. AB - Cardiovascular disease is a major global cause of mortality, and its fundamental underlying substrate is atherosclerosis. Young and old patients have different risk factor profiles, clinical presentations, angiographic findings and prognosis. We performed a retrospective case-control study in a cohort (cases) of premature coronary disease (45 years, n=200). The proportion of premature coronary disease in our geographic area was 9%. The average age of the case group was 41 and 64 years in controls (p<0.001). The male sex, though majority in both groups, was significantly more prevalent in the young group (92.5%) than in the older group (76.0%, p<0.001). The presence of smoking habit, hyperlipidemia and family history was significantly higher in the case group as well, with smoking habit being the most prevalent risk factor. In contrast, hypertension and diabetes were more frequent in controls. The number of affected vessels in cases was significantly less than the control group (1.4+/-0.8 vs. 1.7+/-0.9; p=0.013). Premature coronary disease affects predominantly the male sex and shows high prevalence of cardiovascular risk factors, mainly tobacco, hyperlipidemia, and family history of ischemic heart disease. In addition, it is characterised by a less extensive coronary atherosclerosis, mainly with the higher presence of single-vessel disease in contrast to older patients, as well as lower initial mortality. PMID- 17462763 TI - Unremitting embolus from cardiac myxoma at circumflex artery trifurcation. AB - Embolisation of coronary artery from cardiac myxoma is very rare and it is not clear what happens with embolic material inside coronary artery after myocardial infarction. The natural course of myxomatous embolus is important because it determines the mode of surgical intervention. Different options of the course of embolus have been speculated, from spontaneous resorption to growth at artery wall. We report a case of embolisation of the circumflex artery trifurcation from a villous left atrial myxoma. The course of the embolus was displayed by coronary angiography repeated 6 months after myocardial infarction. Unlike the previously published case report, we found the embolus to be unremitting. PMID- 17462764 TI - Informal care and Medicare expenditures: testing for heterogeneous treatment effects. AB - We estimate the effect of informal care on Medicare expenditures not only for care provided by children but also by the source of informal care (sons versus daughters, children versus others) and recipient characteristics (marital status). Our conceptual framework predicts heterogeneous effectiveness by source and recipient of informal care. We estimate two-part expenditure models as a function of informal care, controlling for endogeneity. We find that informal care by children reduces Medicare long-term care and inpatient expenditures of single elderly. We find that children are less effective caregivers among recipients who are married. For single elderly, child caregivers are more effective than other types. Gender of a child caregiver does not matter. PMID- 17462765 TI - Quantification of bacterial RubisCO genes in soils by cbbL targeted real-time PCR. AB - Soils harbor a high diversity of ribulose-1,5-bisphosphate carboxylase/oxygenase (RubisCO) large subunit coding genes (cbbL). Real-time PCR was used to quantify this gene in differently managed agricultural soils and soil microhabitats. We developed primers and a TaqMan probe that target the "red-like" RubisCO gene cbbL. Primers and probe were developed based on cbbL sequences of selected bacterial pure cultures and of environmental clones. The amount of cbbL copies in the investigated soils were detected in the range of 6.8x10(6) to 3.4x10(7) "red like" cbbL copies/g soil. The cbbL genes could be located entirely in the clay and silt fraction, while the coarse sand fractions revealed no detectable level of bacterial RubisCO genes. These results indicate that bacteria with RubisCO coding genes are numerous and widespread in soils, however the functional implication of this gene in soils is not yet clear. PMID- 17462766 TI - Development of matrix lysis for concentration of gram positive bacteria from food and blood. AB - The development of a fast, reliable and inexpensive protocol for the concentration of bacteria from food by the removal of fat, carbohydrates and proteins that is compatible with downstream alternative DNA-based quantification methods is described. The protocol was used for dairy products, cooked and smoked fish and meat, carbohydrate-rich cooked products, ready-to-eat sauces, egg and blood. Lysis resulted in pellets of reasonable size for further processing. Starch, plant materials, fungi, tissues such as sinew, and chalaza could not be dissolved. Using L. monocytogenes, S. aureus and B. cereus as model organisms, microscopic analysis of the remaining bacterial pellets revealed that the recovered bacteria remained physically intact, albeit that the viability of the cells was compromised. Using real-time PCR, 7.3 CFU of L. monocytogenes were detected in artificially contaminated ultra-high temperature treated (UHT) milk and raw milk. PMID- 17462767 TI - Cochlear expression of a dominant-negative GJB2R75W construct delivered through the round window membrane in mice. AB - Development of a gene-delivery method to the inner ear is an essential step for studies of hearing function and gene therapy. Application of liposomes or adenoviral vectors onto the intact round window membrane (RWM) offers the possibility of atraumatic exogenous gene transfer. GJB2 encodes the gap junction protein Connexin26, which plays a crucial role in potassium recycling in the inner ear. The R75W allele of GJB is a well-characterized mutation that causes deafness at the DFNA3 through a dominant-negative mechanism of action. In this study, a plasmid vector, pGJB2(R75W)-eGFP, was lipocomplexed with N-[1-(2,3 Dioleoloxy)propyl]N,N,N-trimethylammonium methylsulfate: cholesterol and applied onto mouse RWM. At 3 days (3d) post-treatment, immunohistochemistry demonstrated GJB2(R75W)-eGFP transgene expression in the cochlea in: inner and outer pillar cells, outer hair cells, Claudius cells and, in the spiral limbus and ligament. Significant hearing loss was detected by auditory brainstem response testing after 1, 2 and 3d post-treatment; hearing levels returned to control levels at 5d post-treatment. These data confirm that GJB2(R75W) induces functional impairment in the mature cochlea through a dominant negative effect, and importantly, that RWM application of exogenous genes is a feasible method to test their impact on hearing. PMID- 17462768 TI - Myo-inositol hexakisphosphate degradation by Bifidobacterium infantis ATCC 15697. AB - The ability of five strains of different bifidobacterial species (Bifidobacterium animalis, B. bifidum, B. infantis, B. longum, B. pseudolongum) isolated from human and animal gut to degrade myo-inositol hexaphosphate or phytic acid (InsP(6)) has been evaluated. The disappearance of phytate and the generation of lower myo-inositol phosphates were determined in a complex medium in which phytic acid was the only source of phosphorus. Bifidobacterium infantis ATCC 15697 showed the highest level of phytate-degrading activity. This strain displayed optimal activity at slight acid pH (6.0-6.5) and 50 degrees C, but also retained high activity levels at neutral pH and 37 degrees C. B. infantis ATCC 15697 produced maximum activity during the stationary phase of growth and when 1% lactose was used as carbon source. In contrast, the presence of inorganic phosphate in the growth medium inhibited phytase activity. The ability of B. infantis ATCC 15697 to generate lower myo-inositol phosphates and, particularly, accumulate myo-inositol tri-phosphates (InsP(3)) from partial hydrolysis of InsP(6) could contribute to the reduction of the anti-nutritional properties of InsP(6) and the generation of intermediate compounds with beneficial properties during food processing and gastrointestinal transit. PMID- 17462769 TI - Simultaneous quantification of pathogenic Campylobacter and Salmonella in chicken rinse fluid by a flotation and real-time multiplex PCR procedure. AB - A procedure for simultaneous quantification of Campylobacter and Salmonella spp. in poultry skin rinse fluids by a flotation and real-time multiplex PCR method is described. Flotation of the target organisms in a discontinuous density gradient separated them from background microflora, particles from poultry skin, dead target cells and PCR inhibitors. Variation of the buoyant density between 1.052 to 1.106 g/ml was measured at different times for various Salmonella strains grown over a period of 4 weeks. This, and the results from earlier studies on the buoyant densities of Campylobacter spp., which were between 1.065 and 1.109 g/ml, led to design of an optimal discontinuous flotation method with three density layers, of 1.048, 1.109 and approximately 1.200 g/ml. This method preceded a real time multiplex PCR assay using hybridization probes. The specificity of the PCR assay was confirmed on 73 target and non-target strains, and target organisms were detected at the level of one genome per PCR. Results obtained with the combined flotation and real-time multiplex PCR method showed that quantification in rinse fluids was possible down to 3.0+/-0.3 x 10(3) CFU/ml in the presence of other microorganisms at numbers up to 10(9) CFU/ml. PMID- 17462770 TI - Osmotic stress induced by salt increases cell yield, autolytic activity, and survival of lyophilization of Lactobacillus delbrueckii subsp. lactis. AB - Growth and stress adaptation of an autolytic strain of Lactobacillus delbrueckii subsp. lactis FAM-10991 was studied during pH-controlled batch fermentations. After an initial growth to an optical density at 650 nm of 0.8 under controlled optimal growth conditions (pH 5.5, 37 degrees C, no salt), exponentially growing cells were exposed to salt at concentrations from 1 to 3.5%, and temperatures between 48 and 53.5 degrees C, without pH control or with pH controlled at 5.5 or 4.5. Autolysis was induced by salt concentrations of 2.5 or 3.5% and suppressed at 53.5 degrees C or pH 4.5. Salt at concentrations of 2.5 or 3.5% or a temperature of 53.5 degrees C, without pH control or with pH controlled at 5.5, significantly enhanced (p<0.05) survival of lyophilization as compared with the survival of cells in control cultures or cultures with salt at concentration of 1 and 1.5%. The former conditions increased survival by 125- and 200-fold, respectively. However, no correlation was found between autolytic activity and survival of lyophilization. Cultures grown with salt at 2.5% gave high yields of viable cells in broths before and after lyophilization, with numbers being 27 fold higher than with control cultures, but with autolytic activity that was 2.5 fold higher than in cells from control cultures. PMID- 17462771 TI - Autolysis of propionibacteria: detection of autolytic enzymes by renaturing SDS PAGE and additional buffer studies. AB - Five strains of propionibacteria with 70-90% autolysis in sodium lactate broth (SLB) were studied by renaturing sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Several lytic bands ranging in size between 25 and 143 kDa were detected by using propionibacteria cells or cell walls as substrate in the gel. Four Propionibacterium freudenreichii strains showed similar autolytic-enzyme profiles, consisting of two autolytic bands, one with molecular mass 162 kDa and one in the range 123-143 kDa. However, the Propionibacterium acidipropionici strain showed a completely different profile, consisting of 8 autolytic bands with molecular masses of 122, 97, 71, 55, 43, 39, 31, and 25 kDa. Lytic enzymes from P. freudenreichii INF-alpha, P. freudenreichii ISU P-59, P. freudenreichii ISU P-24, and P. freudenreichii ISU P-50 showed lytic activity against cells from all these four strains, but not against P. acidipropionici ATCC 4965. However, P. acidipropionici ATCC 4965 autolysed only its own cells. Effects of pH, temperature, and ions on autolytic activity were tested by renaturing SDS-PAGE and in buffer systems. Results from the SDS-PAGE electrophoresis showed optimal autolytic activity of P. acidipropionici ATCC 4965 at 37 degrees C and in the pH range 7 to 8.5 and of P. freudenreichii ISU P-59 at 20 degrees C and in the pH range 5 to 7. The autolytic activity of P. acidipropionici ATCC 4965 was extremely heat stable (100 degrees C, 2 h), in contrast to the lytic activity of P. freudenreichii ISU P-59, which was heat labile. The autolytic activities of P. acidipropionici ATCC 4965 were inhibited by divalent cations, however, the lytic activities of P. freudenreichii ISU P-59 were activated by Mn(2+), Ca(2+), and Co(2+). In buffer, optimum autolysis of P. acidipropionici ATCC 4965 was observed at pH 8.5 and at 40 degrees C. P. freudenreichii ISU P-59 showed optimum autolysis in buffer at pH 7.5 and at 30 degrees C. PMID- 17462772 TI - Specific Bifidobacterium strains isolated from elderly subjects inhibit growth of Staphylococcus aureus. AB - Cell-free, pH-controlled supernatants of thirty-eight Bifidobacterium strains isolated from healthy elderly subjects were subjected to antimicrobial activity assay. Bioluminescent indicator strains Staphylococcus aureus RN4220, Escherichia coli K-12, and Salmonella enterica serovar Typhimurium ATCC 14028 were used as targets of antimicrobial activity. The effect of nutrient depletion on the inhibition was eliminated with spent-culture controls. Three out of thirty-eight Bifidobacterium strains were capable of inhibiting the growth of S. aureus. The inhibition was equal to 23.2+/-19.1% to 50.4+/-26.7% of the inhibition caused by 50 IU/ml nisin. Reuterin-producing positive strain Lactobacillus reuteri SD2112 was capable of 86.0+/-24.6% inhibition, but Bifidobacterium lactis Bb-12, a known probiotic strain, showed no inhibition. None of the strains was capable of inhibiting the growth of E. coli or S. enterica. The observed inhibition by bifidobacteria was related to hydrogen peroxide formation and possible production of heat-stable proteinaceous compounds. The results suggest that production of antimicrobial substances other than organic acids is not common among Bifidobacterium strains typical of elderly subjects. However, specific strains were identified which showed considerable inhibitory activity against S. aureus. PMID- 17462773 TI - Temperature-assisted pressure inactivation of Listeria monocytogenes in turkey breast meat. AB - Ready-to-eat turkey breast meat samples were surface-inoculated with a five strain cocktail of Listeria monocytogenes cultures to a final concentration of approximately 10(7) CFU/g. The inoculated meat samples were vacuum-packaged and pressure treated at 300 MPa for 2 min, 400 MPa for 1 min, and 500 MPa for 1 min at initial sample temperatures of 1, 10, 20, 30, 40, 50, and 55 degrees C. L. monocytogenes was most resistant to pressure at temperatures between 10 and 30 degrees C. As temperature decreased below 10 degrees C or increased over 30 degrees C, its pressure sensitivity increased. This enhanced inactivation effect was more pronounced when meat samples were treated at higher temperature than at lower temperature. For example, a 1-min treatment of 500 MPa at 40 degrees C reduced the counts by 3.8 log(10), while at 1 and 20 degrees C the same treatment reduced counts by 1.4 and 0.9 log(10), respectively (P<0.05). The survival curves of L. monocytogenes were obtained at 300 MPa and 55 degrees C, 400 MPa and 50 degrees C, and 500 MPa and 40 degrees C. With increasing treatment time, the three survival curves showed a rapid initial drop in bacteria counts with a diminishing inactivation rate or tailing effect. The survival data were fitted with a linear and a nonlinear, Weibull, models. The Weibull model consistently produced better fit to the survival data than the linear model. PMID- 17462774 TI - Optimum bacteriocin production by Lactobacillus plantarum 17.2b requires absence of NaCl and apparently follows a mixed metabolite kinetics. AB - Bacteriocins from lactic acid bacteria are ribosomally synthesized anti-microbial compounds that may find applications from food preservation to healthcare. Food preservation, using in situ bacteriocin production is the most obvious and simple. Frequently, the best conditions for bacteriocin production are those prevailing during food fermentation but a better understanding of the relationship between growth and bacteriocin production is required. In this work, we evaluate the effects of some environmental factors on bacteriocin production by Lactobacillus plantarum 17.2b. A first screening design showed that NaCl, temperature, pH and the type and concentration of carbon and nitrogen sources were most influents. A moderate stimulatory effect of ethanol and oleuropein was also registered. Two consecutive central composite designs were used to examine the effect of the selected variables and to compute its optimum. The evolution of changes produced by the alterations in environmental factors was further examined trough perturbation plots. Bacteriocin production by L. plantarum 17.2b was very sensitive to environmental conditions and uncoupled from growth. Maximum production required suboptimal growth temperatures, pH values above growth's optimum and no NaCl. A preliminary approach to kinetics showed that bacteriocin production by this strain apparently follows mixed metabolite kinetics. PMID- 17462775 TI - Expression of xyloglucan endotransglycosylases of Gerbera hybrida and Betula pendula in Pichia pastoris. AB - The plant enzyme xyloglucan endotransglycosylase (XET; EC 2.4.1.207, xyloglucan:xyloglucosyl transferase) participates in selective modification of plant cell walls during cell growth. XETs are potential catalysts in various applications. Here, sequences encoding two XETs from Gerbera hybrida and Betula pendula are reported. The encoded proteins, which are 51% identical at the amino acid level, were expressed in the yeast Pichia pastoris in secreted form with the aid of mating factor alpha signal sequence. XET production in shake flask cultivations was better at 22 degrees C than at 30 degrees C. Both the yield of protein of expected molecular mass and the XET activity improved at the lower temperature. Under all cultivation conditions studied, higher amounts of XET from B. pendula (BXET) were expressed than XET from G. hybrida (GXET). Both XET enzymes were produced in 16l fed-batch bioreactor cultures. GXET was produced in methanol-limited fed-batch cultivation in minimal medium, and BXET in temperature limited fed-batch (TLFB) in minimal or complex medium. Production was highest in TLFB in complex medium. BXET was purified from the culture filtrate and characterized. Based on the specific activity of the purified protein, 60-70 mg l(-1) BXET was produced in the TLFB in complex medium. PMID- 17462776 TI - Glucose fluctuation on the progression of diabetic macroangiopathy--new findings from monocyte adhesion to endothelial cells. AB - The aim of this study was to elucidate the effect of repetitive fluctuations in blood glucose concentrations on monocyte adhesion to the aortic endothelium. Streptozotocin induced diabetes rats were fed twice daily to induce repetitive postprandial glucose spikes. Then, we compared the number of monocytes adherent to the endothelium of thoracic aorta in these rats with that in rats fed ad libitum. In addition, we evaluated the effect of N-acetyl-L cystein and probucol which are known as antioxidants on the monocyte adhesion to endothelial cells. Streptozotocin induced diabetes rats fed twice daily showed remarkably lower HbA(1c) than the rats fed ad libitum. However, the former group showed markedly higher number of monocytes adherent to the endothelium than the latter. Both N acetyl-L cystein and probucol could not reduce the number of adherent monocytes in streptozotocin induced diabetes rats fed twice daily. Our data demonstrated that repetitive postprandial fluctuation in glucose concentration evokes monocyte adhesion to endothelial cells that was worse than that induced by stable hyperglycemia in vivo. PMID- 17462777 TI - Pathology and pathogenetic mechanisms of diabetic neuropathy: correlation with clinical signs and symptoms. AB - Drastic increase in diabetic patients poses serious problems in the care of neuropathy so that there needs to explore the pathogenesis and to establish the effective treatment. Recent clinical and basic studies revealed characteristic pathophysiology of diabetic neuropathy and some clue to the direction of the treatment. The pathology of diabetic neuropathy is characterized by progressive nerve fiber loss that gives rise to positive and negative clinical signs and symptoms such as pain, paresthesia and loss of sensation. The nerve fiber loss takes the form of pan-modal pattern with proximo-distal gradient. Endoneurial microangiopathic change is also a constant feature of peripheral nerve pathology and negatively correlates with nerve fiber density. The vascular change and distal nerve fiber loss of small caliber, in particular, at the site of epidermis, commence even in subjects with impaired glucose tolerance and precede loss of nerve fibers in the nerve trunk of lower extremities. Pathogenetic mechanisms underlying the progressive nerve fiber loss seem to be multifactorial, including polyol pathway, glycation, reactive oxygen species, and altered protein kinase C activity. Clinical trials based on this background confirmed that fundamental treatment is in fact beneficial for the prevention and halting of this intractable disorder. PMID- 17462778 TI - The depletion of cellular mitochondrial DNA causes insulin resistance through the alteration of insulin receptor substrate-1 in rat myocytes. AB - Since the bioenergetic capacity of skeletal muscle mitochondria is decreased in type 2 diabetes and obesity, the reduction of mitochondrial DNA (mtDNA) content may be involved in the development of insulin resistance in skeletal muscle. To elucidate the association of cellular mtDNA content and insulin resistance, we produced L6 GLUT4myc myocytes depleted of mtDNA by long-term treatment with ethidium bromide (EtBr). L6 GLUT4myc cells cultured with 0.2 microg/ml EtBr (termed depleted cells) revealed a marked decrease in cellular mtDNA, concomitant with a lack of mRNAs encoded by mtDNA. Interestingly, the mtDNA-depleted cells showed a drastic decrease in basal and insulin-stimulated glucose uptake, indicating that L6 GLUT4myc cells develop impaired glucose utilization and insulin resistance. The repletion of mtDNA normalized basal and insulin stimulated glucose uptake. The plasma membrane (PM) GLUT4 in the basal state was decreased, and the insulin-stimulated GLUT4 translocation to the PM was drastically reduced by mtDNA depletion. Interestingly, the expression of IRS-1 associated with insulin signaling was decreased by 90% in the depleted cells, and the insulin-stimulated phosphorylation of IRS-1 and Akt2/PKB were drastically reduced in the depleted cells. Those changes returned to control levels after mtDNA repletion. Taken together, our data suggest that PM GLUT4 content and insulin signal pathway intermediates are modulated by the alteration of cellular mtDNA content, and the reduction in the expression of IRS-1 and insulin stimulated phosphorylation of IRS-1 and Akt2/PKB are associated with insulin resistance in the mtDNA-depleted L6 GLUT4myc myocytes. PMID- 17462779 TI - Short-chain aldehyde-derived ligands for RAGE and their actions on endothelial cells. AB - The formation and accumulation of advanced glycation endproducts (AGE) have been implicated in the development of diabetic vascular complications. Their biological responses are known to be mediated by the receptor for AGE (RAGE). Recently, AGE have been proposed to be derived not only from the classical Maillard reaction but also from other pathways of sugar autoxidation and metabolism. Here, we report the identification of glyceraldehydes (Gcer)- and glycolaldehyde (Gcol)-derived AGE as RAGE ligands and their presence in vivo. The apparent dissociation constants assessed by surface-plasmon resonance (SPR) analysis with purified human RAGE proteins were 360 nM for Gcer-AGE and 1.35 microM for Gcol-AGE. The radiolabeled-ligand binding assay with RAGE-expressing COS-7 cells revealed similar association kinetics. Competitive SPR assay with antibodies specific to the respective AGE fractions demonstrated abundant existence of both Gcer- and Gcol-AGE in RAGE affinity-purified proteins from human sera. The serum contents of Gcer- and Gcol-AGE in a diabetic patient were about twice as high as those in a healthy control. Functionally, Gcer- and Gcol AGE upregulated the endothelial cell levels of mRNA for vascular endothelial growth factor (VEGF) and the secretion of its protein product into the culture media and DNA synthesis in a dose-dependent manner. Further, these endothelial responses were augmented by RAGE overexpression. The results suggest that RAGE engagement of Gcer- and Gcol-AGE may elicit angiogenesis through the induction of autocrine VEGF, thereby contributing to the development and progression of diabetic angiopathies. PMID- 17462780 TI - The modulating effects of the overexpression of uncoupling protein 2 on the formation of reactive oxygen species in vascular cells. AB - Uncoupling protein 2 (UCP-2) is a newly identified member of the mitochondrial anion carrier family and shares 60% sequence identity with the well-characterized thermogenic UCP-1 from brown adipose tissue. Several lines of evidence suggest that UCP-2 is involved in the control of reactive oxygen species (ROS) production by mitochondria. More recently, a direct role for UCP-2 in the regulation of atherogenesis has been suggested by the observation that bone marrow transplantation from UCP-2-deficient mice to low-density lipoprotein receptor deficient mice markedly increased atherosclerotic lesion size. This review introduces the possible role of UCP-2 in the regulation of atherogenesis in vascular cells. Although the relative contribution of the individual ROS generating systems in the vasculature is still ambiguous, both cell membrane NAD(P)H oxidase and the mitochondrial electron-transport chain have been proposed to play significant roles in the overproduction of ROS. UCP-2 can possibly modify atherosclerotic processes initiated in vascular cells and agents that increase UCP-2 expression in vascular cells may help prevent the development and progression of atherosclerosis in patients with diabetes or hypertension. PMID- 17462781 TI - Oral contraception and the risk of hepatocellular carcinoma. AB - BACKGROUNDS/AIMS: We performed a meta-analysis of observational epidemiological studies to examine the association between oral contraceptives (OC) and hepatocellular carcinoma (HCC). METHODS: Two independent researchers conducted PubMed searches followed by systematic abstraction of studies that compared OC use between patients with HCC and a group of controls. Pooling of ORs was conducted using a random effects model. Heterogeneity and publication bias among studies were examined. RESULTS: Twelve case-control studies that included 739 cases and 5223 controls met the inclusion and exclusion criteria. The pooled estimate of ORs (age- and sex-matched only) from all 12 studies was 1.57 (95% CI=0.96-2.54, p=0.07) with a heterogeneity of I(2)=39.9. Exclusion of one large multi-national European study decreased the heterogeneity to I(2)=16.9 and increased the pooled OR to 1.70 (95% CI=1.12-2.59, p=0.01). Eight studies reported adjusted ORs (in addition to age and sex); the pooled estimate was 1.45 (95% CI=0.93-2.27, p=0.11) with a heterogeneity of I(2)=20.4. Only few studies identified or adjusted for other HCC risk factors. Six studies showed a significant 2- to 20-fold increase in HCC risk with longer durations of OC use; however, the reporting was too inconsistent to allow meta-analysis. CONCLUSIONS: The evidence is inconclusive to establish a relation between oral contraceptives and the risk of hepatocellular carcinoma. Future studies should focus on the duration, intermittency, and recency of OC use. PMID- 17462782 TI - Serum adiponectin is increased in advancing liver fibrosis and declines with reduction in fibrosis in chronic hepatitis B. AB - BACKGROUND/AIMS: Despite the possible role of adiponectin in the pathogenesis of liver cirrhosis, few data have been collected from patients in different stages of liver fibrosis. We studied the role of adiponectin in 2 chronic hepatitis B (CHB)-patient cohorts. METHODS: Serum adiponectin was quantified by enzyme-linked immunosorbent assay. One-hundred liver biopsy specimens from CHB patients with different stages of fibrosis and 38 paired liver biopsies from hepatitis B e antigen-positive patients randomized to lamivudine (n=15), pegylated interferon alfa-2a (n=15) or pegylated interferon alfa-2a plus lamivudine (n=8) therapy for 48 weeks were assessed. RESULTS: Serum adiponectin was detected at levels ranging over fourfold magnitude with advancing fibrosis stage and correlated positively with fibrosis stage [r=0.45, p<0.001]. CHB patients with stage 0-1 fibrosis had higher composition of high molecular weight (HMW) form of adiponectin when compared with CHB patients with liver cirrhosis [mean+/-SEM 51.2+/-2.1% vs. 40.9+/-1.7%, respectively, p=0.001]. After antiviral therapy, patients with fibrosis reduction had marked decline in serum adiponectin level and increase in HMW form of adiponectin [mean+/-SEM 43.5+/-1.2% vs. 37.0+/-3.0%, respectively, p=0.04]. CONCLUSIONS: Serum adiponectin may have a role in fibrosis progression in CHB infection. A marked decline in serum adiponectin after antiviral therapy is associated with fibrosis reduction. PMID- 17462783 TI - A transgenic mouse with beta-Galactosidase as a fetal liver self-antigen for immunotherapy studies. AB - BACKGROUND/AIMS: To optimise vaccination strategies for immunotherapy in the liver, we have generated a line of transgenic mice expressing beta-Galactosidase downstream of the alpha-fetoprotein promoter (AFP/betaGal). METHODS: betaGal expression was documented by qRT-PCR, enzyme activity and immunohistochemistry. betaGal-specific CD8+ T-cell activation in mice immunised with various vectors was measured by interferon-gamma ELISpot. RESULTS: Like AFP, betaGal expression was detected in fetal hepatocytes and disappeared around birth. In adult mice, a CD8+ T-cell response to betaGal was observed after immunisation with betaGal adenovirus or plasmid DNA but not with betaGal protein or after retroviral infection. When betaGal was re-expressed in adult hepatocytes, immunisation with betaGal adenovirus triggered T-cell mediated elimination of betaGal-expressing hepatocytes. However, the response was weaker than in AFP/betaGal animals in which betaGal was only present around birth. CONCLUSIONS: In AFP/betaGal mice, betaGal is a fetal liver self-antigen. Interestingly, the basal tolerance to betaGal displayed by these animals is increased during liver re-expression of the self-antigen in adulthood. Adenoviral immunisation allows complete elimination of betaGal-expressing hepatocytes in spite of this increased peripheral tolerance. These results highlight the importance of tolerance against self-antigens and validate the AFP/betaGal mice as a good background to test immunotherapy strategies in hepatocarcinogenesis models. PMID- 17462784 TI - A statistical analysis of pain relief after surgical operations. AB - OBJECTIVES: Administering pain relief measures requires information about development of post-operative pain. Such information can be obtained from studying the patients' perception of pain. The aim of this study is to develop a statistical model that predicts the level of pain in different periods after the operation and the time effects of analgesics. The results can be used for an effective and timely application of pain medications. METHODS: This paper applies an ordered response model to a sample of 392 observations from 49 patients undergoing orthopedic surgeries. The patients' subjective pain levels have been recorded for several intervals up to 24h after their respective operations. The adopted statistical model accounts for the unobserved heterogeneity among patients through random coefficients. Such heterogeneity could be due to differences in patients' perception of pain as well as their health status and sensitivity. RESULTS: The analysis indicates that post-operative pain gradually increases over time but with a slightly diminishing rate. The results suggest that analgesics are quite effective in containing the development of pain. However, the analgesic effects manifest gradually, at a rate more or less similar to that of post-operative pain. CONCLUSION: Highlighting the importance of pain from both clinical and economic perspectives, the analysis indicates that a timely application of analgesics is crucial for an effective pain relief after surgical operations. In particular, the results indicate that the optimal time of administering analgesics is immediately after the operation. The main policy implication of this finding is that preemptive measures of pain relief could be most effective and should be favored to similar analgesic treatments after the manifestation of pain. The results also show that the post-operative pain risks differ significantly across individual cases, which suggests that case-specific clinical assessments supersede any statistical analysis. The proposed statistical model can be used to identify high-risk categories and explore the variations across different analgesic types. PMID- 17462785 TI - A database for medical image management. AB - MEDIMAN (Medical Image MANagement) is a web information system (WIS) for medical image management and processing currently used by neuroscientists and clinicians at several medical and research centres in Spain for research and clinical trials. While developing the MEDIMAN database (DB) we encountered several design challenges unlike those arising in traditional DBs. This paper describes the development of MEDIMAN focusing on the database and the use of the database development process proposed in Midas, a model-driven framework for WIS development. Special attention is given to the design decisions made at each stage to address the challenges encountered. PMID- 17462786 TI - Therapeutic potential for microRNAs. AB - MiRNAs are a conserved class of non-coding RNAs that negatively regulate gene expression post-transcriptionally. Although their biological roles are largely unknown, examples of their importance in cancer, metabolic disease, and viral infection are accumulating, suggesting that they represent a new class of drug targets in these and likely many other therapeutic areas. Antisense oligonucleotide approaches for inhibiting miRNA function and siRNA-like technologies for replacement of miRNAs are currently being explored as tools for uncovering miRNA biology and as potential therapeutic agents. The next few years should see significant progress in our understanding of miRNA biology and the advancement of the technology for therapeutic modulation of miRNA activity. PMID- 17462788 TI - The association between hospitalisation for pneumonia and the diagnosis of lung cancer. AB - Pneumonia may mask or be the first sign of lung cancer. The literature is, however, sparse. We compared the incidence of lung cancer among patients hospitalised for pneumonia with a general population. The patients (index group, N=7044) were hospitalised for pneumonia from 1988 to 2002 at Akershus University Hospital. The reference group was recruited from a general population survey (N=81,373) performed during 1984-1986 and 1995-1997. We had detailed information on smoking habits in both groups. Data on lung cancer incidence, emigration, and death were obtained from the Cancer Registry of Norway. Data were analyzed using multivariate Poisson regression. There were 177 cases of lung cancer in the index group and 608 cases in the reference group. Current smoking modified the association between pneumonia and lung cancer. Among current smokers and non smokers the incidence density ratio (IDR) was 6.0 (95% confidence interval) (4.6 7.8) and 12.0 (8.6-16.9) in the index group compared with the references, respectively. The crude IDR decreased from 78.4 (43.8-146) 0-3 months after the inclusion to 2.4 (1.4-3.7) 6 years after inclusion. In the index group the IDR was only slightly higher for smokers than non-smokers. Patients hospitalised for pneumonia had an increased incidence of lung cancer compared with the general population. This risk persisted for 6 years after the onset of the lung infection. PMID- 17462789 TI - Effect of a moderate exercise on the regulatory hormones of food intake in rats. AB - Strategies used to counteract overweight include generally endurance exercise. Force-resistance exercise has not been tested yet with this objective. The aim of this study was to investigate the response of the main regulatory hormones of food intake (insulin, adiponectin, leptin, ghrelin) and corticosterone, to a short force resistance exercise. Two groups of 16 rats, 65 days old, weighing 330g, were constituted. A standard diet (containing glucid: 72.2, lipid: 7.7, protid: 20% calories) was given "ad libitum". One group served as control, the second group was submitted to exercise training during 5 weeks. Training reduced the rats body weight by 6.4% and the total food intake during the 5 weeks by 11%. Training lowered the insulin and ghrelin levels, while corticosterone level was increased. Insulin, ghrelin and corticosterone only reached the significant threshold p<0.05. Thus, it seems that exercise, even of low intensity and duration, induces changes on hormones that regulate food intake and limit overweight. PMID- 17462790 TI - A novel VIP signaling pathway in T cells cAMP-->protein tyrosine phosphatase (SHP 2?)-->JAK2/STAT4-->Th1 differentiation. AB - Vasoactive intestinal peptide (VIP) is a potent anti-inflammatory agent. In addition to the deactivation of macrophages, dendritic cells, and microglia, VIP shifts the Th1/Th2 balance, promoting the preferential differentiation and survival of Th2 cells, to the detriment of the proinflammatory Th1 effectors. Several mechanisms operate in the Th1/Th2 shift induced by VIP. Here we report on a novel mechanism for the effect of VIP on T cell differentiation, and show that VIP inhibits Th1 differentiation by interfering directly with the IL-12Jak2/STAT4 signaling pathway in T cells. The effect of VIP is cAMP-dependent, and appears to be mediated through the activation of protein tyrosine phosphatases (PTP), with SHP-2 as a potential target. The activation of PTPs represents a novel cAMP downstream target for the immunomodulatory effects of VIP. PMID- 17462791 TI - Evidence that alpha7 nicotinic receptor modulates glutamate release from mouse neocortical gliosomes. AB - The presence of nicotinic receptors on astrocytes in human and rat brain has been previously demonstrated however their possible functional role is still poorly understood. In this study we investigated on the presence of nicotinic receptors on gliosomes, purified from mouse cortex, and on their role in eliciting glutamate release. Epibatidine significantly increased basal release of [3H]D aspartate and of endogenous glutamate from mouse gliosomes but not from synaptosomes. This effect was prevented by methyllycaconitine, alpha-bungarotoxin and mecamylamine but not by dihydro-beta-erythroidine. Epibatidine provoked also a significant increase of calcium concentration in gliosomes but not in synaptosomes; the increase in [Ca2+]i induced by epibatidine and KCl in gliosomes was very similar to each other. The present results indicate that alpha7 nicotinic receptors exist on mouse cortical glial particles and stimulate glutamate release. PMID- 17462792 TI - NT-3 modulates BDNF and proBDNF levels in naive and kindled rat hippocampus. AB - Both mature and precursor forms of neurotrophins regulate nerve development, survival and plasticity. Brain-derived neurotrophic factor (BDNF) synthesis and secretion in turn are regulated by neuronal activity, such as epilepsy. Further, neurotrophins themselves are regulated by neurotrophin levels. Neurotrophin-3 (NT 3) and BDNF in particular can be co-expressed and each can regulate the levels of the other. This regulation is thought to be mediated through receptor tyrosine kinase (Trk) activity. It is not known whether this neurotrophin-neurotrophin interaction occurs in hippocampal tissue in vivo, or how it is influenced by neuronal activation. In this study, we explored the reciprocal influences of intraventricular infusions of NT-3 and BDNF in naive and kindled hippocampi of rats using Western blotting. We confirm that hippocampal kindling resulted in a significant increase in levels of BDNF both in cytochrome C (control) infused and NT-3 infused kindled rats. However, NT-3 infusion significantly reduced BDNF levels in both kindled and non-kindled hippocampi compared to their cytochrome C infused counterparts. These results are consistent with our earlier studies demonstrating lowered levels of TrkA and TrkC (NGF modulates BDNF levels via TrkA) following chronic NT-3 infusion. Although kindling led to an increase in BDNF, this was not accompanied by any detectable change in the levels of proBDNF. However, there was a significant increase in proBDNF following NT-3 infusions, suggesting NT-3 may reduce proBDNF processing. In contrast, neither NT-3 nor proNT-3 levels were affected by kindling or chronic BDNF infusions, consistent with down-regulation of TrkB by chronic BDNF infusion. Thus, modulation of BDNF by NT-3, likely mediated by Trk receptors, occurs in naive and kindled adult rat hippocampus. PMID- 17462793 TI - [Infective endocarditis: epidemiology and diagnosis]. PMID- 17462794 TI - [New classifications and pathophysiology of the inflammatory myopathies]. AB - SCOPE: Review on new classifications of myositis linked with their different pathophysiology. CURRENT SITUATION AND SALIENT POINTS: The classification of myositis refined recently, taking into account clinical (such as isolated muscle involvement or not, association with cancer...), immunological (presence or absence of auto-antibodies) and pathological criteria. This new classification has the ability to separate different clinical and physiopathological entities, having actually different prognosis factors. The most common inflammatory myopathies include dermatomyositis (DM), polymyositis (PM), inclusion body myositis (IBM), but also, overlap myositis (defined, among others, by the presence of auto-antibodies), and myositis associated to cancers. These myopathies may be also distinguished by their histological features which also reflect their different underlying pathogeneses. The mechanism of DM is complement-mediated microangiopathy, the inflammatory infiltrate being secondary to ischaemic damage. In PM the muscle fibres are damaged by cytotoxic CD8 T lymphocytes. IBM may be a degenerative disease with accumulation of a variety of proteins within the fibres. The inflammatory infiltrate, which is similar to that seen in PM, may be secondary to accumulated proteins. PERSPECTIVES: These diseases with different pathogeny and prognosis should be treated by specific approaches. That is the reason why we initiated specific clinical trials for respectively inclusion body myositis and overlap myositis. PMID- 17462795 TI - [Cell therapy for the heart; from bench to bedside]. PMID- 17462796 TI - 137Cs and 40K in the terrestrial vegetation of the Yenisey Estuary: landscape, soil and plant relationships. AB - Plant species, forming important components of Arctic food chains and of interest from a monitoring perspective, were studied at 36 plots representing flood plain and terrace landscapes of the Yenisey River and Estuary from its upper delta to the gulf. (137)Cs contamination densities at the plots varied from 0.35kBq/m(2) (central delta, sandy riverside plot) to 88kBq/m(2) (the upper delta plot) indicating both global and regional sources of anthropogenic pollution. Cs-137 levels in plants were within the range expected from global fallout inputs and varied from 31 to 140Bq/kg d.w. increasing in dominant groups in the order: grassesG sequence variation in the promoters of both HBG1 and HBG2: convergent evolution of the human gamma-globin genes. AB - We encountered a homozygous -16C>G mutation in cis at identical positions in the promoters of both human gamma-globin genes in a subject who was also homozygous for Hemoglobin C (HbC). Subsequent analysis of normal control individuals of African American ancestry revealed that both mutations were always present in cis with an allelic frequency of 3%. Furthermore, 10 out of 11 HbC subjects carried the -16C>G sequence variations, suggesting an association with HbC. The -16C>G mutation disrupts a putative CACCC box positioned between the TATA box and the transcriptional start site. However, the absence of high levels of HbF in HbC subjects homozygous and heterozygous for the -16C>G sequence variation suggested no effect of this mutation on gamma-globin gene expression in the adult stage of development. Further functional characterization by means of transient transfections in human erythroleukemic K562 cells showed that the -16C>G promoter sequence variation did not have an effect on gamma-globin expression in the fetal stage of development either. We therefore conclude that the -16C>G gamma-globin sequence variations are not beneficial to the clinical phenotype of HbC. The unique concurrent presence of this non-functional sequence variation is likely the result of a gene conversion event, and supports the concept of sequence homogenization between the two human gamma-globin genes. PMID- 17462923 TI - A novel gene MGA1 is required for appressorium formation in Magnaporthe grisea. AB - Insertional mutagenesis is an effective way to study the infection mechanism of fungal pathogens. In an attempt to identify the genes involved in appressorium formation from Magnaporthe grisea, we carried out Agrobacterium tumefaciens mediated transformation (ATMT) of the fungus. Analysis of the region flanking the T-DNA integration site in one of the appressorium mutants showed insertion in a gene coding a 78 amino acid protein (MGA1), showing no significant homology to any of the known proteins. The mutant mga1 caused neither foliar nor root infection. Complementation of the mutated gene with the full length wild type gene restored appressorium formation as well as rice infection demonstrating the involvement of this gene in pathogenicity of M. grisea. In an indirect immunolocalisation assay, the MGA1 expression was seen predominantly in germ tube and appressoria. The mutant was impaired in glycogen and lipid mobilization required for appressorium formation. The glycerol content in the mycelia of the mutant under hyperosmotic stress conditions was less as compared to wild type and was thus unable to tolerate the hyperosmotic stress induced by sorbitol. We hypothesize that MGA1 plays a crucial role in signal transduction leading to the metabolism of glycogen and lipids, which is a part of appressorium differentiation process. PMID- 17462924 TI - The gene coding for a new transcription factor (ftf1) of Fusarium oxysporum is only expressed during infection of common bean. AB - We report the isolation and analysis of the gene encoding ftf1 (Fusarium transcription factor 1), a previously undescribed putative transcription factor from highly virulent strains of Fusarium oxysporum f.sp. phaseoli that is transcribed specifically during early stages of infection of its host common bean (Phaseolus vulgaris L.). The predicted 1080 amino acid ftf1 protein contains a Zn(II)2-Cys6 binuclear cluster DNA-binding motif. ftf1 expression during axenic growth in culture was not detected by either Northern or RT-PCR. On the contrary, in planta transcription of ftf1 is increased about 24h after plant inoculation, as detected by real-time RT-PCR. This result suggests that ftf1 has a role in the establishment of the fungus within the plant and/or the progress of the disease. Multiple copies of ftf1 are present in highly virulent strains of F. oxysporum f.sp. phaseoli. PMID- 17462925 TI - Transverse oriented electric field re-entrant resonator (TERR) with automatic tuning and coupling control for EPR spectroscopy and imaging of the beating heart. AB - Sample motion, particularly that of a beating heart, induces baseline noise and spectral distortion on an EPR spectrum. In order to quench motional noise and restore the EPR signal amplitude and line-width, an L-band transverse oriented electric field re-entrant resonator (TERR) was designed and constructed with provisions for automatic tuning control (ATC) and automatic coupling control (ACC) suited for studies of isolated beating rat hearts. Two sets of electronic circuits providing DC biased voltage to two varactor diodes were implemented to electronically adjust coupling and tuning. The resonator has a rectangular cross sectional sample arm of 25 mm diameter with a Q value of 1100 without sample. Once inserted with lossy aqueous samples of 0.45% NaCl, Q value drops to 400 with a volume of 0.5 ml and 150 with 5 ml. The ATC/ACC functions were tested with a moving phantom and isolated beating rat hearts with the improvement of signal to noise ratio (S/N, peak amplitude of signal over peak amplitude of baseline noise) of 6.7-, and 4 to 6-fold, respectively. With these improvements, EPR imaging could be performed on an isolated beating rat heart. Thus, this TERR resonator with ATC/ACC enables application of EPR spectroscopy and imaging for the measurement and imaging of radical metabolism, redox state, and oxygenation in the isolated beating rat heart. PMID- 17462926 TI - Effects of beta-carotene on adult immune condition and antibacterial activity in the eggs of the Grey Partridge, Perdix perdix. AB - Carotenoids are important dietary constituents in birds. Their functions are numerous and complex, and breeding females are potentially faced with an optimal allocation of these resources between themselves and offspring. We conducted a dietary experiment (low and high supply of beta-carotene) to examine the effect of beta-carotene on health and immune response of 64 reproducing pairs of Grey Partridge (Perdix perdix L.) and on the quality of their eggs, as revealed by the measurement of biochemical components in yolk and albumen, the egg hatching rate and chick survival. We found a beneficial effect of beta-carotene on the erythrosedimentation rate and immune response of females (PHA reaction), while the diet did not significantly affect these variables in males. In both sexes, the plasma level of carotenoids was not related to the quantity of beta-carotene supplied. A higher quantity of beta-carotene in the diet did not induce a variation of egg nutrients (proteins and lipids), nor an increase of yolk beta carotene concentration. We detected a higher concentration of lysozyme, an enzyme with antibacterial activity, in the albumen of eggs laid by females with a high supply of beta-carotene. These eggs showed higher hatching rates. The present study indicates that although carotenoid supplementation does not influence blood and yolk carotenoid levels, it results in better immune conditions of females, eventually translated into increased antibacterial activity of the eggs. The broad range of beneficial effects of carotenoids is discussed. PMID- 17462927 TI - Calcium ATPase expression in the oviducts of the skink, Lampropholis guichenoti. AB - Lampropholis guichenoti is an oviparous lizard that lays eggs with a calcareous outer shell. We used immunofluorescence microscopy to describe the occurrence and distribution of Ca2+ ATPase pumps in the uterus of L. guichenoti at different stages of the reproductive and egg-shelling cycles. Ca2+ ATPase pumps were not demonstrated by immunofluorescent techniques in any uterine tissue until egg shelling had commenced and at least partly calcified eggs were in the uterus. During egg-shelling, Ca2+ ATPase pumps occur on the apical and baso-lateral surfaces of uterine epithelial cells, and those of associated shell glands in the stroma of the uterus. We conclude that Ca2+ ATPase pumps provide a major mechanism for deposition of the calcareous eggshell of L. guichenoti and that the pumps are up-regulated when required in the reproductive cycle. Furthermore, it is likely that specific calcium glands in the stroma of the uterus are involved in the rapid transport required for egg-shelling, but the differential contribution of luminal and glandular epithelial cells is not known. PMID- 17462928 TI - Sleep and wakefulness in the green iguanid lizard (Iguana iguana). AB - The reptile Iguana iguana exhibits four states of vigilance: active wakefulness (AW), quiet wakefulness (QW), quiet sleep (QS) and active sleep (AS). Cerebral activity decreases in amplitude and frequency when passing from wakefulness to QS. Both parameters show a slight increase during AS. Heart rate is at a maximum during AW (43.8+/-7.9 beats/min), decreases to a minimum in QS (25.3+/-3.2 beats/min) and increases in AS (36.1+/-5.7 beats/min). Tonical and phasical muscular activity is present in wakefulness, decreases or disappears in QS and reappears in AS. Single or conjugate ocular movements are observed during wakefulness, then disappear in QS and abruptly reappear in AS. Although these reptiles are polyphasic, their sleep shows a tendency to concentrate between 20:00 and 8:00 h. Quiet sleep occupies the greater percentage of the total sleep time. Active sleep episodes are of very short duration, showing an average of 21.5+/-4.9 (mean+/-SD). Compensatory increment of sleep following its total deprivation was significant only for QS. Reaction to stimuli decreased significantly when passing from wakefulness to sleep. It is suggested that the lizard I. iguana displays two sleep phases behaviorally and somatovegetatively similar to slow wave sleep and paradoxical sleep in birds and mammals. PMID- 17462929 TI - Distribution and characteristics of endogenous digestive enzymes in the red-eared slider turtle, Trachemys scripta elegans. AB - Distribution and properties of the main digestive enzymes including protease and amylase, from stomach, pancreas and the anterior, middle and posterior intestine of the adult red-eared slider turtle Trachemys scripta elegans were studied at various pHs and temperatures. The optimum temperature and pH for protease in stomach, pancreas and the anterior, middle and posterior intestine were 40 degrees C, 2.5; 50 degrees C, 8.0; 50 degrees C, 7.0; 50 degrees C, 8.0; and 50 degrees C, 8.5; respectively. The optimum temperature and pH for amylase in stomach, pancreas and anterior, middle and posterior intestine were 40 degrees C, 8.0; 30 degrees C, 7.5; 40 degrees C, 7.0; 50 degrees C, 8.0; and 50 degrees C, 8.0; respectively. Under the optimum conditions, the order of protease activity from high to low was of pancreas, stomach and the anterior, posterior and middle intestine; the activity of amylase in descending order was of anterior intestine, pancreas, posterior intestine, middle intestine and stomach. PMID- 17462930 TI - Osmotic stress and muscle tissue volume response of a freshwater bivalve. AB - The freshwater bivalve, Corbicula fluminea, when submitted to hyperosmotic solutions, behaves as a hyperosmoconformer; we have observed an increase in osmolality and ions in its extracellular fluid. Osmotic and ionic changes in its watery environment represent a challenge for the tissues of this mollusk. Thus we evaluated, in vitro, muscle tissue volume variations (based on wet weight change) under anisosmotic salines, as well the possible regulatory mechanisms involved in the processes. This tissue did not exhibit complete volume regulation under anisosmotic saline solutions, but showed less variation than would be predicted by Van't Hoff's law, and tissue volume remained essentially stable throughout 90 min of exposure. To minimize tissue swelling in hyposmotic situations, C. fluminea muscle mobilizes organic osmolytes (ninhydrin positive substances) and inorganic ions (K(+) and Cl(-)). While under hyperosmotic stimulus, apparently only inorganic osmolytes (Na(+) and Cl(-)) are mobilized by the tissue. Our results indicate ionic accumulation by the Na(+)-K(+)-2Cl(-) cotransporter and the Na(+)/H(+) coupled to Cl(-)/HCO(3)(-) exchangers. Exposure of the muscle tissue to Ca(2+)-free anisosmotic saline did not result in a detectable inhibition of the mechanisms described above. The Ca(2+) gradient that derives from the absence of this ion, even apparently enhances the regulatory mechanisms. These responses of this freshwater mollusk in hyperosmotic solutions, and the muscle tissue under anisosmotic (hypo and hyperosmotic) saline solutions, have not been previously characterized in the manner and approach as reported here. Specifically, we analyze both organic and inorganic osmolytes mobilized under hyposmotic stress, and can infer the participation of Na(+) and Cl(-) pathways stimulated by hyperosmotic stress. From the perspective gained in this study, tissue volume responses may be used as models for toxicological investigations. PMID- 17462931 TI - Ribosomal protein L24 is differentially expressed in ovary and testis of the marine shrimp Marsupenaeus japonicus. AB - In order to identify genes involved in oogenesis in shrimp, an ovarian cDNA library of Marsupenaeus japonicus was screened using a suppression-subtraction hybridization (SSH)-enriched probe. More than 20 genes were identified as differentially expressed genes between the ovary and the testis. Unexpectedly, one of these genes is a ribosomal protein that is normally considered a housekeeping gene. Northern blot shows that the shrimp ribosomal protein L24 gene (srpl24) is 0.6 kb in length. The expression level of srpl24 in the ovary is much higher than in the testis. Bioinformatics analyses show that srpl24 encodes a protein of 164 aa with a predicted molecular mass of 18.2 kDa, which is a cytoplasmic ribosomal protein. Real time PCR analyses demonstrated that the relative abundance of srpl24 mRNA in the different organs is: ovary >> testis, hepatopancreas, muscle and eye. The highest expression level of srpl24 in the ovary suggests that srpl24 has an important role in oogenesis. It is the first reported rpl24 in crustaceans and is the first reported rpl24 that is differentially expressed between the ovary and the testis in animals. PMID- 17462932 TI - Concentration of carotenoids, retinol and alpha-tocopherol in plasma of six microchiroptera species. AB - To adequately feed species in captivity it is necessary to know their nutritional habits and their natural availability of specific nutrients. Such essential nutrients are vitamin A, vitamin E and selected carotenoids as vitamin-A precursors. Because their blood plasma concentration are valid biomarkers of nutritional status of dietary intake, we determined the concentrations of carotenoids, retinol and alpha-tocopherol by HPLC as well as the transport proteins for retinol, the retinol-binding protein (RBP) and transthyretin (TTR) immunologically in the plasma of six species of microchiroptera from free-ranging animals and compared it in one species (Carollia perspicillata) to a group held in captivity. Plasma concentrations of the investigated components were generally much lower compared to most other mammals. Within the bats, differences were observed for all components. As in other species retinol, RBP and TTR were present but no retinyl esters could be detected. Plasma of the insectivorous bat species Molossus molossus contained carotenoids. Within the group of carotenoids, beta-carotene was dominant and only traces of lutein were present. Phyllostomus hastatus revealed the highest alpha-tocopherol concentration. No differences in the plasma content of the investigated compounds were found between a group of Carollia perspicillata kept in captivity for 20 years and free-ranging individuals from a population in Central America. No sex related differences were obvious. In conclusion, nutritional biomarkers in bats were highly variable due to dietary and possible species-specific differences. PMID- 17462933 TI - Effect of thermal stress on protein expression in the mussel Mytilus galloprovincialis Lmk. AB - The exposure of organisms to stressing agents may affect the level and pattern of protein expression. Certain proteins with an important role in protein homeostasis and in the tolerance to stress, known as stress proteins, are especially affected. Different tissues and cells show a range of sensitivities to stress, depending on the habitat to which organisms have adapted. The response of different tissues and cells from the mussel Mytilus galloprovincialis Lmk. to heat shock has been studied in this work using different exposure times and temperatures. During the assays, protein expression was observed to vary depending on the tissue studied, the temperature or the exposure time used. But maybe the most prominent thing is the different response obtained from the cultured haemocytes and those freshly obtained from stressed mussels, which makes us think that the extraction procedure is the main cause of the response of non cultured cells, although the haemolymph may contain components that modulate haemocyte response. PMID- 17462934 TI - Short stature and decreased insulin-like growth factor I (IGF-I)/growth hormone (GH)-ratio in an adult GH-deficient patient pointing to additional partial GH insensitivity due to a R179C mutation of the growth hormone receptor. AB - OBJECTIVE: Genetic factors play an expanding role in understanding growth hormone (GH) disorders, therefore the German KIMS Pharmacogenetics Study was initiated with the aim of genotyping various GH-/IGF-I-axis-related genes of GH-deficient adult patients to investigate genotype:phenotype relationships and response to GH therapy. PATIENTS AND METHODS: 129 consecutively enrolled GH-deficient adult patients were genotyped for variant 1 (V1) of the alternatively spliced noncoding exons in the 5'-untranslated region and for the nine coding exons of the GH receptor (GHR) gene, which obviously play a striking role in the function of the GH-IGF-I-axis. After detection of a heterozygous, non-synonymous mutation R179C in exon 6 in one single patient with acquired GH-deficiency (GHD) in late adulthood, analysis of her clinical data followed, leading to the diagnosis of mild short stature (-1.5SD). For further endocrine evaluation, five pituitary stimulation tests (arginine) of this patient were statistically compared to stimulation tests (arginine) of ten GH-deficient control patients, retrospectively. RESULTS: The formerly in patients with Laron syndrome and idiopathic short stature reported mutation R179C leads to an amino acid change from an arginine residue (codon CGC) to a cysteine residue (codon TGC) in position 179 of the extracellular domain of the GHR. Statistical analysis revealed significant decreased IGF-I/GH(0) ratio (p=0.004) and IGF-I/GH(max) ratio (p=0.001) of the index patient compared to the control patients, implying growth hormone resistance of the index patient at the level of the GHR, according to the detected R179C mutation. CONCLUSIONS: This study reports on the unusual case of a patient with mild short stature, who acquired GHD in late adulthood due to a non-secreting pituitary adenoma and get additionally diagnosed for pre existing growth hormone insensitivity due to a formerly in two short statured patients described, single, heterozygous, non-synonymous mutation in the GHR. Our findings support the theory that heterozygous mutations in the GHR gene can have mild phenotypical consequences. PMID- 17462935 TI - Glucocerebrosidase mutations in Chinese subjects from Taiwan with sporadic Parkinson disease. AB - BACKGROUND: An association between glucocerebrosidase, the enzyme deficient in Gaucher disease, and the synucleinopathies has been suggested both by the development of parkinsonism in Gaucher probands and carriers, as well as by the presence of mutations in the gene for glucocerebrosidase (GBA) in different series of subjects with synucleinopathies. In this study, an open access Parkinson repository was used to establish the incidence of GBA alterations in a different ethnic cohort with sporadic Parkinson disease (PD). METHODS: The glucocerebrosidase gene was sequenced in samples collected from 92 Chinese Parkinson disease patients from Taiwan along with 92 clinically screened controls, matched for age and ethnicity. FINDINGS: The frequency of GBA mutations among the Chinese PD probands was 4.3%, in contrast to 1.1% in Chinese controls. Mutant alleles identified included two known mutations, L444P and D409H, and two novel mutations, L174P and Q497R. INTERPRETATION: These results, ascertained in subjects from Taiwan collected in a standardized and clinically rigorous open access Parkinson disease repository and screened by direct sequencing of GBA, demonstrate that GBA mutations are also encountered in Chinese subjects with sporadic PD at a higher frequency than many other known PD genes. The study demonstrates that the association of GBA mutations with the development of parkinsonian pathology is not related to ethnic origin. PMID- 17462936 TI - Handling of human short-chain acyl-CoA dehydrogenase (SCAD) variant proteins in transgenic mice. AB - To investigate the in vivo handling of human short-chain acyl-CoA dehydrogenase (SCAD) variant proteins, three transgenic mouse lines were produced by pronuclear injection of cDNA encoding the wild-type, hSCAD-wt, and two disease causing folding variants hSCAD-319C>T and hSCAD-625G>A. The transgenic mice were mated with an SCAD-deficient mouse strain (BALB/cByJ) and, in the second generation, three mouse lines were obtained without endogenous SCAD expression but harboring hSCAD-wt, hSCAD-319C>T, and hSCAD-625G>A transgenes, respectively. All three lines had expression of the transgene at the RNA level in liver, muscle or brain tissues. Expression at the protein level was detected only in the brain tissue of hSCAD-wt mice, but here it was significantly higher than the level of endogenous SCAD protein in control mouse brains--in correlation with expression at the RNA level. The results may indicate that the two hSCAD folding variants are degraded by the mouse mitochondrial protein quality control system. Indeed, pulse-chase studies with isolated mitochondria revealed that soluble variant hSCAD protein was rapidly eliminated. This is in agreement with the fact that no disease phenotype developed for any of the lines transgenic for the hSCAD folding variants. The indicated remarkable efficiency of the mouse protein quality control system in the degradation of SCAD folding variants should be further substantiated and investigated, since it might indicate ways to prevent disease causing effects. PMID- 17462937 TI - [Point about DNA profiling: technologies, applications, and legislation]. AB - Molecular biology progress in DNA fingerprinting has revolutionized forensic science. It's a reliable tool to identify an individual; it provides a true "genetic identity card". It's based of the concept that no two individuals can have an identical DNA pattern except identical twins. This article is a clarification on current technologies used in DNA profiling, their applications and its legislation, according a special place to the national databases (FNAEG), which is in great development for the moment. PMID- 17462938 TI - [ABO incompatibility and non myeloablative allogeneic stem cell transplantation]. AB - ABO incompatibility is not a barrier for allogeneic hematopoietic stem cell transplantation but is associated with specific complications. Major ABO incompatibility is associated with delayed erythroid engraftment, increased transfusion requirement and cases of pure red cell aplasia. Minor ABO incompatibility may be responsible for acute haemolytic reactions in the first months following transplantation. The widely used non myeloablative conditioning regimens might modify the management of ABO incompatibility. They could favour pure red cell aplasia development in the setting of major ABO mismatch since they are associated with a prolonged persistence of host anti-donor isohemagglutinins after allogeneic hematopoietic stem cell transplantation. In the setting of minor ABO incompatibility, the use of peripheral blood stem cells and the nature of graft-versus-host disease prophylaxis regimen may have an impact on the incidence of haemolytic reactions. In that review, the clinical and therapeutic aspects of ABO incompatibility are studied, especially regarding the impact of the conditioning regimen intensity. PMID- 17462939 TI - Sonoluminescence, sonochemistry (H2O2 yield) and bubble dynamics: frequency and power effects. AB - In the present work, comprehensive experimental and numerical investigations on the effects of ultrasound frequency and acoustic power on sonoluminescence (SL) and H(2)O(2) yields have been carried out. The multibubble SL and H(2)O(2) yields have been examined for four frequencies (213, 355, 647 and 1056 kHz) and over a wide range of acoustic powers. The observed experimental results have been discussed with respect to single bubble dynamics and the number of active cavitation bubbles. PMID- 17462940 TI - Synthesis and characterization of copper oxide (I) nanoparticles produced by pulsed sonoelectrochemistry. AB - Cu(2)O nanopowders have been prepared by ultrasound-assisted electrochemistry with a potentiostatic set-up. Their composition has been determined by X-ray diffraction and energy dispersive X-ray spectroscopy. Transmission electron microscopy and centrifugation analyses indicate that the nanopowders consist of agglomerates of variable nanometric diameter grain. Most of particles have a diameter of 8 nm whatever the electrodeposition potential. The influence of the parameters of electrochemical and ultrasonic pulses on the particle diameter was also studied. The specific surface areas determined by Brunauer-Emmet-Teller (BET) model are very high with a value close to 2000 m(2)g(-1). PMID- 17462942 TI - Predominant expression of lysosomal N-acylethanolamine-hydrolyzing acid amidase in macrophages revealed by immunochemical studies. AB - Bioactive N-acylethanolamines, including anandamide (an endocannabinoid), N palmitoylethanolamine (an anti-inflammatory substance), and N-oleoylethanolamine (an anorexic substance) are enzymatically hydrolyzed to fatty acids and ethanolamine. Fatty acid amide hydrolase plays a major role in this reaction. In addition, we cloned cDNA of an isozyme termed "N-acylethanolamine-hydrolyzing acid amidase (NAAA)" [K. Tsuboi, Y.-X. Sun, Y. Okamoto, N. Araki, T. Tonai, N. Ueda, Molecular characterization of N-acylethanolamine-hydrolyzing acid amidase, a novel member of the choloylglycine hydrolase family with structural and functional similarity to acid ceramidase, J. Biol. Chem. 280 (2005) 11082-11092]. Previous biochemical analyses suggested the expression of NAAA in macrophage cells and various rat tissues including lung and brain. To clarify the physiological significance of NAAA, here we immunochemically studied NAAA for the first time. We developed an antibody specific for rat NAAA, and by Western blotting revealed that NAAA is glycosylated and subjected to specific proteolysis. In alveolar macrophages isolated from rat lung, NAAA was immunocytochemically localized in lysosomes. In the whole lung tissue, only alveolar macrophages were immunostained for NAAA. Conformably, the mRNA and protein levels and activity of NAAA in alveolar macrophages were much higher than those in the whole lung tissue. In brain, intraventricular macrophages were positively stained with anti-NAAA antibody, while microglia appeared to be negative. These results strongly suggested the importance of macrophages as an expression site of NAAA in rat tissues. PMID- 17462943 TI - Short latency afferent inhibition is not impaired in mild cognitive impairment. AB - OBJECTIVE: To determine whether cortical cholinergic circuit impairment exists in the mild cognitive impairment (MCI) brain. METHODS: Fifteen healthy elderly controls (NC), 16 amnesic MCI subjects and 12 probable Alzheimer's disease (AD) subjects were recruited. Conditioning stimuli were delivered at the right wrist followed by test transcranial magnetic stimulation (TMS) of the left motor cortex. The center of the linear contiguous segment of the coil was placed over a point 5 cm lateral to the vertex on the interaural line. The interstimulus intervals (ISIs) between the conditioning stimuli and the test stimuli were set at 20, 40, 100, 200 and 600 ms. An inhibitory effect that occurred at ISIs as short as 20 ms was defined as short-latency afferent inhibition (SAI). RESULTS: SAI was significantly reduced in subjects with AD compared with NC, but it was not reduced in subjects with MCI. CONCLUSIONS: A difference in cortical excitability between subjects with AD and subjects with MCI could be captured by an in vivo neurophysiological method. SIGNIFICANCE: The state of the neurotransmitter systems, including the cortical cholinergic system, is thought by some compensatory mechanisms to be kept at the normal level in subjects with MCI. PMID- 17462944 TI - Free copper and resting temporal EEG rhythms correlate across healthy, mild cognitive impairment, and Alzheimer's disease subjects. AB - OBJECTIVE: The present study tested the hypothesis that the serum copper abnormalities were correlated with alterations of resting electroencephalographic (EEG) rhythms across the continuum of healthy elderly (Hold), mild cognitive impairment (MCI), and AD subjects. METHODS: Resting eyes-closed EEG rhythms delta (2-4Hz), theta (4-8Hz), alpha 1 (8-10.5Hz), alpha 2 (10.5-13Hz), beta 1 (13 20Hz), beta 2 (20-30Hz), and gamma (30-40Hz), estimated by LORETA, were recorded in 17 Hold, 19 MCI, 27 AD- (MMSE< or =20), and 27 AD+ (MMSE20) individuals and correlated with copper biological variables. RESULTS: Across the continuum of Hold, MCI and AD subjects, alpha sources in parietal, occipital, and temporal areas were decreased, while the magnitude of the delta and theta EEG sources in parietal, occipital, and temporal areas was increased. The fraction of serum copper unbound to ceruloplasmin positively correlated with temporal and frontal delta sources, regardless of the effects of age, gender, and education. CONCLUSIONS: These results sustain the hypothesis of a toxic component of serum copper that is correlated with functional loss of AD, as revealed by EEG indexes. SIGNIFICANCE: The present study represents the first demonstration that the fraction of serum copper unbound to ceruloplasmin is correlated with cortical delta rhythms across Hold, MCI, and AD subjects, thus unveiling possible relationships among the biological parameter, advanced neurodegenerative processes, and synchronization mechanisms regulating the relative amplitude of selective EEG rhythms. PMID- 17462945 TI - Late auditory evoked potentials asymmetry revisited. AB - OBJECTIVE: To investigate brain asymmetries of the auditory evoked potential (AEP) N100, T-complex, and P200 in response to monaural stimulation. METHODS: Electroencephalographic (EEG) recordings from 68 channels were used to record auditory cortex responses to monaural stimulation from normal hearing participants (N=16). White-noise stimuli and 1000Hz tones were repeatedly presented to either the left or right ear. Source localization of the AEP N100 response was carried out with two symmetric regional sources placed into left and right auditory cortex. Regional source waveform amplitude and latency asymmetries were analyzed for tangential and radial activity explaining the N100, T-complex and P200 AEP components. RESULTS: Regional source waveform analysis showed that early tangential activity in the N100 latency range exhibited larger contralateral amplitudes and shorter latencies for both tone and noise monaural stimuli. Lateralized activity was significantly greater when tones or noise was presented to the left compared to the right ear (p<.001). The ear difference in the degree of lateralization arose due to hemispheric asymmetry. Significantly more tangential activity in the N100 latency range was recorded in the right compared to the left hemisphere in response to stimulation by either tones or noise (p<.001). Neither the radial activity modelling the T-complex, nor activity modelling the P200, showed robust ear or hemisphere differences. CONCLUSIONS: Regional source waveform analysis revealed that the extent of auditory evoked potential asymmetries depends on the ear and hemisphere examined. These findings have implications for future studies utilizing AEP asymmetries to examine normal auditory function or experience-related changes in the auditory cortex. SIGNIFICANCE: The right compared to the left auditory cortex may be more involved in processing monaurally presented tone and noise stimuli. PMID- 17462946 TI - Right ventricular dysfunction as an independent predictor of short- and long-term mortality in patients with heart failure. AB - BACKGROUND: The prognostic importance of right ventricular (RV) dysfunction in heart failure (HF) has been suggested in patients with severe systolic heart failure. Tricuspid annular plane systolic excursion (TAPSE) is a simple echocardiographic measure of RV ejection fraction, but may be affected by co existing chronic obstructive pulmonary disease (COPD). AIMS: To examine the prognostic information from TAPSE adjusted for the potential confounding effects of co-existing cardiovascular and COPD in a large series of patients admitted for new onset or worsening HF. METHODS AND RESULTS: Eight hundred and seventeen patients screened for participation in a large clinical trial by trans-thoracic echocardiography, including measurement of TAPSE, were followed for a median of 4.1 years (maximum 5.5 years). Decreased TAPSE as well as presence of COPD were independently associated with adverse short- and long-term survival, hazard ratio was 0.74 (p=0.004) for every doubling of TAPSE; and 2.4 (p<0.0001) for the presence of COPD. CONCLUSION: Decreased RV systolic function as estimated by TAPSE is associated with increased mortality in patients admitted for HF, and is independent of other risk factors in HF including left ventricular function. The co-existence of COPD is also associated with an adverse prognosis independent of the RV systolic function. PMID- 17462947 TI - Angiotensin-converting enzyme inhibitors and survival in women and men with heart failure. AB - BACKGROUND: Several randomized controlled trials demonstrate that angiotensin converting enzyme (ACE) inhibitors improve survival in patients with congestive heart failure (CHF). However, whether ACE inhibitors benefit both sexes is not adequately addressed. PURPOSE: Our objective was to determine the effectiveness of ACE inhibitors in women with CHF. METHODS: The Quebec hospital discharge database was linked with the physician and drug claims database to identify a cohort with a discharge diagnosis of CHF between January 1998 and March 2003. In this retrospective cohort study, subjects who filled a prescription for ACE inhibitors (19,220 exposed) were compared to those who never filled such prescription (8617 non-exposed). The primary outcome was survival by exposure to ACE inhibitors. MAIN FINDINGS: There were 14,693 women (67% exposed) and 13,144 men (72% exposed). The 1 year mortality was 19.5% and 30% in those exposed and non-exposed, respectively. A significant survival benefit was demonstrated in both sexes exposed to ACE inhibitors [adjusted hazard ratio (95% confidence interval): women 0.80 (0.76-0.85); men 0.71 (0.67-0.75)]. PRINCIPAL CONCLUSIONS: ACE inhibitors improve survival in both sexes with CHF, but the protective effect appears to be greater in men. Our results support the current recommendations for the management of women with CHF. PMID- 17462948 TI - Intrathoracic impedance monitoring to detect chronic heart failure deterioration: relationship to changes in NT-proBNP. AB - BACKGROUND: An alert algorithm, based on intrathoracic impedance monitoring, has been incorporated into a cardiac resynchronisation device (CRT) to detect pulmonary fluid accumulation, and to audibly alert patients to decompensating chronic heart failure (CHF). AIMS: To evaluate this algorithm, alert events were correlated with changes in NT-proBNP concentration and CHF status. METHODS AND RESULTS: In a prospective observational study of 62 patients (89% male, aged 67+/ 1 year), NT-proBNP plasma concentrations, clinical CHF status, and device data were collected at enrolment, during regular follow-up and at device alerts. Over a mean follow-up of 27+/-2 weeks, pooled data indicated a weak, but significant inverse relationship between relative changes in intrathoracic impedance and NT proBNP (r=-0.3; p<0.001). In 52 device alerts from 35 patients, NT-proBNP increased by 66+/-19% from 2039+/-331 pg/ml (p<0.001). The increase in NT-proBNP was higher in alerts with clinical signs of CHF deterioration (n=30, 89+/-25%; p<0.001) than in alert events without clinical signs (n=22, 25+/-15%; p=n.s.). CONCLUSION: Intrathoracic impedance based alert events are associated with a significant increase in NT-proBNP concentration. These data indicate that intrathoracic impedance monitoring might facilitate the outpatient management of CHF patients with implanted CRT devices. PMID- 17462949 TI - Yersinia outer proteins E, H, P, and T differentially target the cytoskeleton and inhibit phagocytic capacity of dendritic cells. AB - Through Yersinia outer proteins (Yops) Yersinia disrupt the actin cytoskeleton of epithelial cells and macrophages, and this leads to a decreased capability of these cells to internalize bacteria. We examined the effects of different Yops of Y. enterocolitica serotype O8 on the cytoskeleton and phagocytic capacity of murine dendritic cells (DCs). DCs were infected with several Yersinia mutant strains deficient in one Yop or translocating only a single Yop. Analyses of infected DCs by microscopy showed that YopE, YopH and YopT cooperate to rapidly damage the actin cytoskeleton of DCs. Furthermore, microscopic analyses and gentamicin killing assays revealed that the maximum reduction of bacterial uptake was achieved by Yersinia mutant strains translocating only a single Yop (YopE or YopH) indicating that these Yops enable Yersinia to inhibit the phagocytic function of DCs. PMID- 17462951 TI - Aspects of genome plasticity in pathogenic Escherichia coli. AB - The species Escherichia coli comprises not only non-pathogenic or commensal variants that belong to the normal intestinal flora of most mammals, but also various pathogenic strains causing diverse intestinal and extraintestinal infections in man and animals. Virulence factors and mechanisms involved in pathogenesis have been successfully analyzed for many years resulting in a wealth of knowledge about many E. coli pathotypes. However, our knowledge on the genome content, diversity and variability between pathogenic and also non-pathogenic subtypes is only slowly accumulating. Pathotypes have been largely defined by the presence or absence of particular DNA segments that in most cases appear to have been acquired via horizontal gene transfer events. As these regions are frequently subjected to excisions, rearrangements, and transfers they contribute to the previously unexpected and underestimated rapid evolution of E. coli variants resulting in the development of novel strains and even pathotypes. In these studies various novel aspects of genome diversity and plasticity in extraintestinal and intestinal pathogenic E. coli pathotypes have been addressed and the results have been directly applied for the improvement of diagnostic methods. PMID- 17462952 TI - Memories are made of this: synergy of T cell receptor and cytokine signals in CD4(+) central memory cell survival. AB - The ability of the immune system to 'remember' a previous encounter with antigen is the hallmark of the adaptive immune response. The signals required for CD4(+) memory T cell survival are not well understood, and a recent paper lends new biochemical insights into how memory T cells might optimally be preserved through the convergence of signals from the engagement of cytokine and antigen-specific T cell receptors. PMID- 17462953 TI - A key role for CCR7 in establishing central and peripheral tolerance. AB - Early studies identified the CC-chemokine receptor (CCR)7 as an important homing molecule controlling the lymph node entry of naive T cells through high endothelial venules and of activated mature dendritic cells through afferent lymphatics. Consequently, these properties initially branded CCR7 as a central organizer of the primary immune response. However, recent studies have demonstrated that a variety of immune cells crucially rely on CCR7-directed migration not only for the induction of protective immunity but also for the establishment of immunological tolerance. In this review, we therefore highlight some of the recent advances in understanding the multiple roles of CCR7 in the induction and maintenance of central and peripheral tolerance. PMID- 17462954 TI - VEGF-targeted cancer therapy strategies: current progress, hurdles and future prospects. AB - Despite setbacks, the clinical development of antiangiogenic agents has accelerated remarkably over the past 3-4 years. Consequently, there are currently three direct inhibitors of the VEGF pathway approved for use in cancer therapy. Other agents that block the VEGF pathway are in advanced stages of clinical development and have shown promising results. With these exciting developments come crucial questions regarding the use of these new molecular-targeted agents, alone or in combination with standard cytotoxic or targeted agents. Importantly, the mechanisms of action of anti-VEGF therapy remain unknown. Here, we discuss several potential mechanisms of action such as tumor vascular normalization, bone marrow-derived cell recruitment blockade and cytostatic effects of anti-VEGF therapy. We review the current progress, the major stumbling blocks and the future directions for anti-cancer therapy using anti-VEGF agents, emphasizing clarification of the underlying molecular mechanisms of action and biomarker identification and validation. PMID- 17462955 TI - Response to Cox and Abeku: Early warning systems for malaria in Africa: from blueprint to practice. PMID- 17462956 TI - Decreased blood dendritic cell counts in type 1 diabetic children. AB - In this study DC numbers, phenotype and DC responses to the Toll-like receptor (TLR)-3 ligand, poly I:C, were examined in new-onset Type 1 diabetes (T1D) patients (ND) and in established T1D patients (ED). Absolute blood myeloid DC (MDC) and plasmacytoid DC (PDC) numbers were decreased in ND and ED patients compared to age-matched controls. The decrease in MDC and PDC counts was less evident in patients with a combination of T1D and coeliac disease (CD) or CD alone. The age-dependent decline in blood DC numbers, found in control children, was not evident in ND patients, such that 2-10 years old ND children had similar MDC and PDC numbers to 15-17 years old controls. In ED patients the t-score of MDC and PDC numbers related to the age of diagnosis but not to disease duration. Blood DC in T1D patients were not distinguished from those of controls by the levels of HLA-DR, CD40 and CD86 expression or the percentage of DC expressing cytokines, IL-12, IL-10, IL-6 and TNF-alpha, in responses to poly I:C. If low DC numbers are shown to contribute to the autoimmunity in T1D, interventions aimed to increase DC numbers may mitigate against beta-cell loss. PMID- 17462957 TI - Risk screening for spontaneous preterm labour. AB - Prevention of viable spontaneous preterm birth before 34 weeks' gestation through screening is one of the key aims of antenatal care as birth below this threshold of prematurity has implications for child, mother and society. If women can be identified to be at high risk of spontaneous preterm birth in early pregnancy, they can be targeted for more intensive antenatal surveillance and prophylactic interventions (primary prevention). However, the disease mechanisms behind preterm birth are not well understood. Consequently, tests for its prediction and treatments for its prevention are not well developed. Additionally, no randomised controlled trial focusing on prevention of spontaneous preterm birth related perinatal morbidity and mortality through a screening programme exists. This chapter describes a generic framework for combining screening information with therapeutic effect to delineate its role in a screening programme. We use test treatment combination of previous history of preterm birth and progestational agents as an example. A decision-making framework is built using: (1) evidence for post-test probabilities; (2) evidence for therapeutic effectiveness; and (3) integration of the two evidences to estimate the effect of the test-treatment combination with numbers needed to treat (NNTs). The NNT to prevent one case of spontaneous preterm birth before 34 weeks' gestation with progesterone is seven in women with a previous history; NNT is 41 in women without a previous history; and it is 28 when previous history was not used to guide a decision about prevention. The proposed framework makes decisions about screening and prevention explicit. PMID- 17462958 TI - The use of contrast echocardiography for the detection of cardiac shunts. AB - Recently, debate has erupted about the clinical significance of cardiovascular shunts. Several major health problems such as stroke and migraine have been associated with patent foramen ovale (PFO) with right-to-left shunt (RLS). The nature of the relationship between these syndromes and PFO is not clearly understood. Technical advances have led to more therapeutic options including device closure of PFO, hence prevention of such a PFO-related stroke has become feasible. Therefore, optimal diagnosis of PFO has become of greater clinical importance. Contrast echocardiography with non-transpulmonary contrast agents has been the cornerstone in diagnosis of PFO with RLS for over four decades. Despite being a relatively invasive procedure, transesophageal echocardiography (TEE) is considered the gold standard for detection of RLS. Several other echocardiographic techniques such as transthoracic echocardiography (TTE) with second harmonic imaging and transcranial Doppler ultrasonography (TCD) have shown increased sensitivity and specificity compared to TEE for the detection of PFO with RLS. Moreover, improvement of skills and techniques used for detection of these shunts has led to greater detection of small and large sized RLS in the echocardiographic laboratory. This review gives and overview of the echocardiographic techniques, contrast agents and manoeuvres used for detection of the major cardiovascular shunts and their clinical relevance to major health problems. PMID- 17462959 TI - Pain and suffering as viewed by the Hindu religion. AB - Religion and spiritual practices are among the resources used by patients to cope with chronic pain. The major concepts of Hinduism that are related to pain and suffering are presented. Ways that Hindu traditions deal with pain and suffering are reviewed, including the concept of acceptance, which has been studied in the pain medicine literature. By becoming more familiar with Hindu views of pain and suffering, pain medicine practitioners can offer potentially helpful concepts to all patients and support Hindus' spirituality as it relates to pain and suffering. PERSPECTIVE: Religion or spirituality is often important to patients. This article will inform the pain medicine practitioner how pain and suffering are viewed in Hinduism, the third largest religion in the world. It is hoped that these concepts will prove helpful when treating not only followers of Hinduism but all patients. PMID- 17462960 TI - Self-efficacy correlates with leg muscle pain during maximal and submaximal cycling exercise. AB - Quadriceps muscle pain is a naturally occurring consequence of moderate- to high intensity cycle ergometry. The present study involved an examination of self efficacy for tolerating muscle pain during exercise as a correlate of muscle pain during maximal and submaximal cycle ergometry. Young adult females (n=16) who were healthy and regularly physically active completed a measure of self-efficacy for tolerating moderate to strong pain in the legs and then undertook a maximal incremental exercise test on a cycle ergometer. Within 1 week, participants completed a 30-minute bout of submaximal exercise (80% VO2peak) on a cycle ergometer. Ratings of quadriceps muscle pain intensity were recorded every minute during the maximal incremental exercise test and every 5 minutes during the submaximal exercise session. Self-efficacy for tolerating pain was moderately inversely correlated with ratings of peak muscle pain during the maximal incremental exercise test (r=-.45) and pain ratings averaged across the submaximal session (r=-.49). Our results are consistent with social-cognitive theory and indicate that self-efficacy for tolerating pain is inversely associated with ratings of muscle pain during maximal and submaximal exercise in healthy and regularly active young adult females. Such findings support a possible examination of strategies for manipulating self-efficacy for tolerating pain and thereby reducing muscle pain during exercise. PERSPECTIVE: Our data provided evidence that self-efficacy for tolerating moderate to strong muscle pain during exercise correlates with muscle pain intensity ratings during bouts of maximal and submaximal exercise. Future research could identify methods of manipulating self-efficacy as a means of decreasing muscle pain during exercise and ultimately enhancing physical activity participation. PMID- 17462961 TI - Development, implementation, and a cognitive evaluation of a definitional question answering system for physicians. AB - The published medical literature and online medical resources are important sources to help physicians make patient treatment decisions. Traditional sources used for information retrieval (e.g., PubMed) often return a list of documents in response to a user's query. Frequently the number of returned documents from large knowledge repositories is large and makes information seeking practical only "after hours" and not in the clinical setting. This study developed novel algorithms, and designed, implemented, and evaluated a medical definitional question answering system (MedQA). MedQA automatically analyzed a large number of electronic documents to generate short and coherent answers in response to definitional questions (i.e., questions with the format of "What is X?"). Our preliminary cognitive evaluation shows that MedQA out-performed three other online information systems (Google, OneLook, and PubMed) in two important efficiency criteria; namely, time spent and number of actions taken for a physician to identify a definition. It is our contention that question answering systems that aggregate pertinent information scattered across different documents have the potential to address clinical information needs within a timeframe necessary to meet the demands of clinicians. PMID- 17462962 TI - Genetic diversity and major spoligotype families of drug-resistant Mycobacterium tuberculosis clinical isolates from different regions of Turkey. AB - To highlight the transmission rate and major phylogenetic clades of drug resistant Mycobacterium tuberculosis isolates, a total of 200 drug-resistant strains isolated in four different regions of Turkey (Marmara n=81; Mediterranean n=39; Aegean n=42; East Anatolia n=38), were typed by spoligotyping and IS6110 restriction fragment length polymorphism (RFLP). The major spoligotyping-defined shared-types (STs) and corresponding lineages were, ST 41 (22.5%, LAM7-TUR), ST53 (19.5%, ill-defined T super-family), ST 50 (6.5%, Haarlem 3), ST 1261 (4.5%, LAM7 TUR), ST 47 (3.5%, Haarlem 1), as well as two STs that belonged to undefined clades (ST 284, 3%, and ST 2067, 2.5%). The global distribution of major M. tuberculosis lineages among drug-resistant strains was as follows: T super-family (29%), Latin-American & Mediterranean (33.5%), Haarlem (14%), and the S lineage (3%). A high number of strains (n=29, 14.5%) showed patterns that did not fall within major clades described so far. A combination of spoligotyping and IS6110 RFLP fingerprinting methods resulted in a final clustering rate of 38.5% and a recent transmission rate of 25.5%. Our results underline the highly diverse nature of drug-resistant tuberculosis in our study population, as well as its ongoing transmission with lineages that are specific to these regions, the most predominant being the LAM7-TUR lineage which shows an enhanced phylogeographical specificity for Turkey. PMID- 17462963 TI - Study of the adsorption and oxidation of antioxidant rutin by cyclic voltammetry voltabsorptometry. AB - The adsorption and oxidation behavior of rutin was studied by in-situ UV spectroelectrochemistry in a long optical-path thin-layer electrochemical cell with a graphite-wax electrode. The dynamic UV spectra of rutin under potentiostatic oxidation were recorded, which indicated the formation of o quinonic structure. During the repetitive cyclic potential scans, cyclic voltabsorptomogram was recorded at the three characteristic wavelengths 346, 254 and 296 nm, respectively. The profiles obtained showed two types of concentration fluctuation of species in solution, resulting from adsorption/desorption and redox reaction, respectively. Using derivative cyclic voltabsorptometry the contribution of the species in solution to the total current was estimated, and then the current of every step involved in the proposed redox mechanism was obtained for the first time. The result shows that rutin underwent a nearly reversible redox reaction in which the total current is mostly due to the contribution of adsorbed species. The present work developed cyclic voltabsorptometry into a useful tool for investigating redox process involving coupled adsorption/desorption steps. PMID- 17462964 TI - Impact of intertrigeminal region AMPA receptor blockade on respiratory responses in rats. AB - We hypothesized that functional glutamatergic AMPA receptors are expressed in the intertrigeminal region (ITR) of the lateral pons, and that blockade of these receptors by a specific AMPA receptor antagonist (NBQX) would alter respiratory responses both to ITR glutamate injections as well as to vagally mediated reflex apnea induced by intravenous infusion of serotonin. Non-selective blockade of ITR glutamate receptors using kynurenic acid previously was shown to alter both of these respiratory responses. Unilateral ITR injections in 19 anaesthetized spontaneously breathing adult Sprague-Dawley rats with the vagi intact demonstrated that NBQX (10mM): (1) shortened glutamate-induced apnea duration (p=0.006), (2) reduced glutamate-induced apnea frequency (p=0.034) and (3) decreased glutamate-induced apnea density (p=0.006). The same dose of NBQX did not affect vagally mediated reflex apnea induced by intravenous infusion of serotonin. These results show that functional glutamate AMPA receptors are expressed in the ITR but fail to directly demonstrate any specific physiologic role for these receptors in respiratory control. In contrast to antagonism of NMDA receptors which completely blocked the central apnea evoked by glutamate, antagonism of AMPA receptors only partially blocked the effects of glutamate. In contrast to kynurenic acid, antagonists to NMDA and AMPA receptors given separately did not potentiate the duration of reflex apnea. PMID- 17462965 TI - Simultaneous determination of alpha-lipoic acid and its reduced form by high performance liquid chromatography with fluorescence detection. AB - The simultaneous determination of alpha-lipoic acid (LA) and DHLA (reduced form of LA) was carried out by HPLC with fluorescence detection. DHLA in the sample was first labeled with ABD-F at room temperature for 10 min and then the LA was labeled with SBD-F at 50 degrees C for 1 h after conversion to DHLA using the reducing agent, TCEP. The resulting fluorophores, ABD-DHLA and SBD-DHLA, were separated by reversed-phase chromatography and detected at 510 nm (excitation at 380 nm). Both fluorophors were completely separated without any interference of endogenous thiols and disulfides in the sample and sensitively detected by fluorimetry. The proposed method was applied to the assay of the LA supplement and the determination in human plasma after the oral administration of LA tablets. The concentration (%) of LA in the tablet was reasonable to the stated amount. Furthermore, the result of a time course study in the plasma after the administration of LA did not differ from a previous report. Thus, the present method seems to be applicable to the simultaneous determination of LA and DHLA in various biological specimens. PMID- 17462966 TI - On-line clean-up and screening of oxacillin and cloxacillin in human urine and plasma with a weak ion exchange monolithic column. AB - A weak ion exchange monolithic column prepared by modifying the GMA-MAA-EDMA (glycidyl methacrylate-methacrylic acid-ethylene glycol dimethacrylate) monoliths with ethylenediamine was applied to remove matrix compounds in biological fluid. Using this monolithic column, on-line clean-up and screening of oxacillin and cloxacillin in human urine and plasma samples had been investigated. Chromatography was performed by reversed-phase HPLC on a C(18) column with ultraviolet detection at 225 nm. Results showed that the ion exchange monolithic column could be used for deproteinization and retaining oxacillin and cloxacillin in human urine and plasma, which provided a simple and fast method for assaying drugs in human urine and plasma. PMID- 17462967 TI - Lupus mastitis in male mimicking a breast lump. PMID- 17462968 TI - Validation of risk assessment scoring systems for an audit of elective surgery for gastrointestinal cancer in elderly patients: an audit. AB - The goal of this study was to validate the usefulness of risk assessment scoring systems for a surgical audit in elective digestive surgery for elderly patients. The validated scoring systems used were the Physiological and Operative Severity Score for enUmeration of Mortality and morbidity (POSSUM) and the Portsmouth predictor equation for mortality (P-POSSUM). This study involved 153 consecutive patients aged 75 years and older who underwent elective gastric or colorectal surgery between July 2004 and June 2006. A retrospective analysis was performed on data collected prior to each surgery. The predicted mortality and morbidity risks were calculated using each of the scoring systems and were used to obtain the observed/predicted (O/E) mortality and morbidity ratios. New logistic regression equations for morbidity and mortality were then calculated using the scores from the POSSUM system and applied retrospectively. The O/E ratio for morbidity obtained from POSSUM score was 0.23. The O/E ratios for mortality from the POSSUM score and the P-POSSUM were 0.15 and 0.38, respectively. Utilizing the new equations using scores from the POSSUM, the O/E ratio increased to 0.88. Both the POSSUM and P-POSSUM over-predicted the morbidity and mortality in elective gastrointestinal surgery for malignant tumors in elderly patients. However, if a surgical unit makes appropriate calculations using its own patient series and updates these equations, the POSSUM system can be useful in the risk assessment for surgery in elderly patients. PMID- 17462969 TI - Modern advances in the understanding of bone structure. PMID- 17462970 TI - How molecular biology can improve clinical management: the MammaPrint experience. PMID- 17462971 TI - The puzzle of genomics. PMID- 17462972 TI - Molecular biology of gastric cancer. AB - Despite its decreasing incidence overall, gastric cancer is still a challenging disease. Therapy is based mainly upon surgical resection when the tumour remains localised in the stomach. Conventional chemotherapy may play a role in treating micrometastatic disease and is effective as palliative therapy for recurrent or advanced disease. However, the knowledge of molecular pathways implicated in gastric cancer pathogenesis is still in its infancy and the contribution of molecular biology to the development of new targeted therapies in gastric cancer is far behind other more common cancers such as breast, colon or lung. This review will focus first on the difference of two well defined types of gastric cancer: intestinal and diffuse. A discussion of the cell of origin of gastric cancer with some intriguing data implicating bone marrow derived cells will follow, and a comprehensive review of different genetic alterations detected in gastric cancer, underlining those that may have clinical, therapeutic or prognostic implications. PMID- 17462973 TI - Genomic instability: on the birth and death of cancer. AB - The presence of an abnormal chromosomal content is probably the most universally conserved hallmark of cancer cells. Predicted at the beginning of the 20th century as the origin of tumours, and extensively documented thereafter, genomic instability lies at the core of neoplastic development. Regardless of this classic model, the actual impact that deficient control of genomic integrity has on human health and particularly on cancer development only started to gain attention from the scientific community two decades ago. From a bird's eye view and with a cancer-oriented perspective, in this work we will try to cover some of the concepts obtained from recent research in genomic instability. The review will end up presenting suggestive evidence which proposes that genomic instability might turn out to be not just the driving force but also the Achilles' heel of cancer. PMID- 17462974 TI - Yeast on drugs: Saccharomyces cerevisiae as a tool for anticancer drug research. AB - The budding yeast Saccharomyces cerevisiae is being widely used as a model for investigating fundamental processes relevant to all living organisms. Many of these processes are affected by genetic and epigenetic alterations in cancer such as cell cycle progression, DNA replication and segregation, maintenance of genomic integrity and stress responses. Therefore, yeast emerges as an attractive model for anticancer drug research. The genetic tractability of budding yeast, its ease of manipulation and the wealth of functional genomics tools available in this organism makes it ideal for genome-wide analysis of biological functions and chemical screenings. The present review will discuss some of the innovative advantages based on yeast genetics and genomics for antitumour drug target identification and drug discovery. PMID- 17462975 TI - A revision on cryoglobulinaemia associated to neoplastic diseases. AB - Cryoglobulinaemia is an infrequent process and has been classified in types I, II and III. Type I corresponds to monoclonal cryoglobulinaemia; types II and III are comprised in the group of mixed cryoglobulinaemias (MC). In MC type II, cryoglobulinaemias are made up of a monoclonal rheumatoid factor vs. a polyclonal IgG. In MC type III all compounds are polyclonal. Monoclonal cryoglobulinaemias are usually associated to malignant haematological processes, while MC are associated to systemic diseases and infection. More research is needed on the natural history, causes, treatment and evolution of cryoglobulinaemia. This revision analyses a more recent knowledge on this entity, as well as its association to neoplastic diseases. PMID- 17462976 TI - Germ cell tumours of the ovary. AB - Germinal cell tumours represent only 2-5% of all cancers of the ovary. However, the characteristics of the tumour and the patients have some special qualities as high rates of healing goes together with a strong desire to keep fertility intact because this condition occurs in female children and adolescent girls. Neither the prognosis nor the treatment of these tumours is homogeneous; the low incidence is the reason it is hard to develop prospective studies for establishing prognostic factors and specific treatments. The introduction of adjuvant chemotherapy into initial surgery has improved the prognosis of these patients. The BEP scheme has proved to be equally efficient and less toxic than PVB, and for this reason it has become the standard scheme despite the fact that no randomised studies have been carried out. The surgical treatment demands the application of the same principles seen in cytoreduction surgery of epithelial cancers of the ovary (maximum possible cytoreduction), though in many cases hysterectomy and double adnexectomy may be obviated. In view of the rarity of these tumours, it is advisable to work within cooperative groups that may have subgroups for the treatment of rare tumours. This will probably be the only way to move forward in the prospective knowledge of prognostic factors for these tumours. PMID- 17462977 TI - Phase II trial: concurrent radio-chemotherapy with weekly docetaxel for advanced squamous cell carcinoma of head and neck. AB - INTRODUCTION: Standard fractionation radiation therapy (RT) combined with concomitant chemotherapy (CT) based on cisplatin schemes is actually the standard treatment for locally advanced non-resectable squamous cell carcinoma of head and neck (SCCHN). The appearance of taxoids has introduced a new kind of treatment with high antitumoral power. The aim of this study is to add more information about the role of this new approach. MATERIALS AND METHODS: Twenty-six patients with locally advanced non-resectable SCCHN were recruited at six institutions in Spain, between January 2001 and January 2003. Docetaxel was administered weekly, for 6 weeks, concurrently with RT. RESULTS: The mean total delivered dose of RT was 70'2 Gy (range 64-74 Gy). The median and mean duration of time were 63 days and 61 days (range 49-103 days) respectively. After a median time control of 19 months (range 3.3-42.2 months), the response rate was 83.4%. The median time to local progression was 16.4 months (95% confidence interval [CI]=4.4-28.4 months). The median survival time was 26.9 months, with one- and two-year overall survival of 66.9% (95% CI=48.1-85.7%) and 57.5% (95% CI=37.3-77.7%) respectively. The median duration time response was 15.1 months (95% CI=3.7-26.5 months). The median time until treatment failure was 9.4 months (95% CI=4.7-14.1). Incidence of grade III-IV mucositis was 88%, neutropenia 72% and skin toxicity 92% (24% grade III-IV). The incidence of severe late toxicity (grade III and IV) due to RT/CT was 31.4%. CONCLUSIONS: Although therapeutics results are equivalent to cisplatin schemes of concurrent CT-RT, mucositis and cutaneous toxicity registered in this trial must be considered as limiting factors to application of this new approach. PMID- 17462978 TI - Low dose rate brachytherapy in lip carcinoma. AB - BACKGROUND: Lip cancer is frequently treated with surgery although radiation therapy offers comparable results. The aim of the study was to evaluate the local cure rate in patients with lip carcinoma treated with 192-Ir low dose rate interstitial brachytherapy. METHODS: Fifty-four patients with a mean age of 70 years (range, 40-90 years) were retrospectively evaluated. The tumour location was the superior lip in 4 (7.4%) and the inferior lip in 50 (92.6%). Tumour stage was T1N0 in 33 patients and T2N0 in 21 patients. The radioactive sources with hypodermic needles in 49 patients (90.7%) and plastic tubes in 5 (9.3%) were placed parallel and equidistant from one another across the tumour volume according to the Paris system rules. RESULTS: The median dose was 61.5 Gy (range, 60-65 Gy). All patients experienced acute brisk skin and mucositis RTOG grade III around the implanted volume, subsiding within 4-6 weeks after the implant. Local control was achieved in 98% of patients. The mean follow-up was 7 years. CONCLUSIONS: Low dose rate interstitial brachytherapy with 192-Iridium is a well established and efficacious way to achieve local control of the tumour in lip cancer. It offers the advantage of avoiding surgery in an elderly population. PMID- 17462979 TI - Disseminated classic Kaposi's sarcoma. AB - Kaposi's sarcoma (KS) is characterised by proliferation of vascular endothelial and lymphoreticular cells, frequently with a multicentric expression developed from a single node and evolving to multiple cutaneous lumps or plaque-like appearance. Four types of KS with similar histological patterns have been described in terms of their clinical and epidemiological features: classic KS, endemic (African) KS, iatrogenic KS and epidemic (AIDS-related) KS. The differences in clinical features are quite relevant: classic KS is usually limited to the lower extremities; whereas immunodeficiency-related diseases frequently involve several organs. A case of a 67-year-old woman with metastatic KS and unproven immunodeficiency is presented. PMID- 17462980 TI - Breast malignant phyllodes tumour metastasising to soft tissues of oral cavity. AB - Metastases from infraclavicular neoplasms to the oral cavity are rare, and the anatomical structures most frequently affected are the maxillae. The primary sites that develop metastases to head and neck include lung, breast, liver, thyroid gland, kidney and melanomas, among others. The breast is one of the usual primary regions that can present them. In the literature, most metastases to these regions correspond to squamous cell carcinomas and adenocarcinomas; few studies report other type of neoplasms such as phyllodes tumour (PT) that present this behaviour. The importance of identifying metastatic disease to the oral cavity, be it as the first manifestation of a neoplasm from unknown origin or as a development in the course of the disease, is that it allows determination of adequate treatment, which has an effect on the patient's prognosis. The usual behaviour of PT is frequent recurrence and, when metastases are present, these are to lungs and bone. Two cases of malignant PT metastasising to tongue and lip are presented, as well as a review of the literature. PMID- 17462981 TI - Successful treatment with vincristine in PHACES syndrome. AB - We report a case of PHACES syndrome witch was effectively treated using vincristine. A female infant was referred at 8 months of age for evaluation with a segmental mandibular haemangioma with rapid growing and facial disfigurement. The infant was initially placed on oral prednisone with clinical response but she developed obstructive sleep apnoea. Vincristine was started at about 24 months and continued for 4 months with marked decrease in the size of the haemangioma. The only side effect was constipation. PMID- 17462982 TI - Parotid gland metastasis of a breast cancer. AB - Parotid gland metastases from malignant tumors are extremely rare. A 61-year-old woman was diagnosed with an early breast cancer with no expression of oestrogen and progesterone receptors. Five years later the patient presented a tumour in parotid gland. After total parotidectomy, microscopic analysis of the gland demonstrated an invasive duct carcinoma (IDC) with positive expression of oestrogen receptor. The patient was treated with chemotherapy followed by complementary local radiotherapy. Diagnosis of a metastasic tumour in parotid gland poses a challenge. In our case an immunohistochemical study of oestrogen receptor was fundamental to establish a diagnosis. PMID- 17462983 TI - Family planning advice and postpartum contraceptive use among low-income women in Mexico. AB - CONTEXT: In Mexico, family planning advice has been incorporated into the clinical guidelines for prenatal care. However, the relationship between women's receipt of family planning advice during prenatal care and subsequent contraceptive use has not been evaluated. METHODS: Data were collected in 2003 and 2004 in 17 Mexican states from 2,238 urban low-income women postpartum. Participating women reported on prenatal services received and contraceptive use. Logistic and multinomial logistic regression models evaluated whether receiving family planning advice during prenatal care predicted current contraceptive use, after quality of care in the community, service utilization, delivery characteristics, household socioeconomic characteristics, and maternal and infant characteristics were controlled for. RESULTS: Overall, 47% of women used a modern contraceptive method. Women who received family planning advice during prenatal care were more likely to use a contraceptive than were those who did not receive such advice (odds ratio, 2.2). Women who received family planning advice had a higher probability of using condoms (relative risk ratio, 2.3) and IUDs (5.2), and of undergoing sterilization (1.4), than of using no method. CONCLUSIONS: Integrating family planning advice into prenatal care may be an important strategy for reaching women when their demand for contraception is high. PMID- 17462984 TI - Factors affecting vasectomy acceptability in Tanzania. AB - CONTEXT: Calls for increased inclusion of men in matters of reproductive health emphasize the need for research into vasectomy acceptability and decision making. Vasectomy is a safe, simple and effective method of contraception, but is underused worldwide. METHODS: Focus group discussions and in-depth interviews were conducted with potential and actual sterilization clients and their partners in the Kigoma Region of Tanzania. Content analysis was used to search for emergent themes related to vasectomy decision making. RESULTS: Six themes emerged as overarching factors contributing to the vasectomy decision-making process: economics, spousal influence, religion, provider reputation and availability, uncertainty about the future, and poor vasectomy knowledge and understanding. There was substantial communication between partners regarding the vasectomy decision, and wives had a strong influence on the outcome; however, men and women agreed that husbands would resist vasectomy if wives initially raised the topic. Vasectomy acceptance is limited by the scarcity of skilled vasectomy providers and by the fact that men and women hold many of the same misunderstandings about vasectomy, including a fear of decreased sexual performance as a result of the procedure. CONCLUSIONS: Spousal discussions are important in the decision to get a vasectomy, but these discussions should be initiated by the male partner. Programs need to educate men about contraceptive options, including vasectomies. Detailed, culturally relevant knowledge of the barriers and facilitators individuals experience during their decision- making process will enable vasectomy promotion programs to more successfully target appropriate populations. PMID- 17462985 TI - Family planning programs in 2004: new assessments in a changing environment. AB - CONTEXT: Periodic assessments between 1972 and 1999 found consistent increases in the intensity and types of effort exerted by national family planning programs in developing countries. An updated evaluation was needed to examine whether these trends have been affected by recent changes in the family planning environment, such as decentralization, the HIV/AIDS pandemic and funding reductions. METHODS: In 2004, informants in 82 developing countries completed a questionnaire that assessed 30 dimensions of program effort and included several new scales to explore current issues. Selected results were compared with findings from prior rounds of the study. RESULTS: Family planning effort increased between 1999 and 2004, both globally and within regions. When the data were weighted by country population size, effort declined slightly overall but increased in four of six regions. Countries with low initial scores improved more than those with high initial scores. Contraceptive access varied by region and was lowest in Sub Saharan Africa. The strongest justifications for programs were improving maternal and child health and preventing unwanted births. Changes in funding were often judged to have had negative effects on programs. Unmarried youth and women receiving postabortion care received the least emphasis among special populations of interest. CONCLUSIONS: Although average program effort scores have risen again, increases in effort, funding and access to contraceptive methods are still needed in many countries, especially in rural areas and among the poor. More emphasis should be placed on providing postpartum and postabortion family planning services. PMID- 17462986 TI - Inconsistencies in the relationship between contraceptive use and fertility in Bangladesh. AB - CONTEXT: Contraceptive prevalence increased by nine percentage points from 1993 to 2000 in Bangladesh, but there was almost no decline in the total fertility rate. METHODS: Data from the 1999-2000 Bangladesh Demographic and Health Survey and from the Matlab Demographic Surveillance System area collected between 1978 and 2001 were analyzed to explain the lack of change in fertility and to examine relationships among contraceptive prevalence, the abortion ratio, desired fertility and total fertility. RESULTS: After a maternal and child health and family planning program was initiated in part of Matlab in 1977, the total fertility rate in the intervention area declined from 4.8 in 1979 to 2.9 in 2000, while fertility in the comparison area dropped from 6.3 to 3.5. Over this period, contraceptive prevalence rose from 30% to 70% and from 16% to 50% in the two areas, respectively; meanwhile, the abortion ratio fell from 4.3 to 3.6 in the intervention area, but rose from around two to 8.2 in the comparison area. Trends in desired fertility in each area were similar, declining from about 4.0 children per woman in 1979 to about 2.5 children in 2000. Among women at each level of parity, fertility generally decreased as the number of sons increased, and fertility was highest for women without sons. CONCLUSIONS: Preference for male children and parental concern over infant and child mortality may partially explain the difference between desired family size and fertility. A reduction in breast-feeding and an increase in use of less-effective contraceptive methods might be responsible for the inconsistency in the relationship between contraceptive use and fertility. PMID- 17462987 TI - Ajulemic acid, a synthetic nonpsychoactive cannabinoid acid, bound to the ligand binding domain of the human peroxisome proliferator-activated receptor gamma. AB - Ajulemic acid (AJA) is a synthetic analog of THC-11-oic acid, a metabolite of tetrahydrocannabinol (THC), the major active ingredient of the recreational drug marijuana derived from the plant Cannabis sativa. AJA has potent analgesic and anti-inflammatory activity in vivo, but without the psychotropic action of THC. However, its precise mechanism of action remains unknown. Biochemical studies indicate that AJA binds directly and selectively to the isotype gamma of the peroxisome proliferator-activated receptor (PPARgamma) suggesting that this may be a pharmacologically relevant receptor for this compound and a potential target for drug development in the treatment of pain and inflammation. Here, we report the crystal structure of the ligand binding domain of the gamma isotype of human PPAR in complex with ajulemic acid, determined at 2.8-A resolution. Our results show a binding mode that is compatible with other known partial agonists of PPAR, explaining their moderate activation of the receptor, as well as the structural basis for isotype selectivity, as observed previously in vitro. The structure also provides clues to the understanding of partial agonism itself, suggesting a rational approach to the design of molecules capable of activating the receptor at levels that avoid undesirable side effects. PMID- 17462988 TI - Arabidopsis VTC2 encodes a GDP-L-galactose phosphorylase, the last unknown enzyme in the Smirnoff-Wheeler pathway to ascorbic acid in plants. AB - The first committed step in the biosynthesis of L-ascorbate from D-glucose in plants requires conversion of GDP-L-galactose to L-galactose 1-phosphate by a previously unidentified enzyme. Here we show that the protein encoded by VTC2, a gene mutated in vitamin C-deficient Arabidopsis thaliana strains, is a member of the GalT/Apa1 branch of the histidine triad protein superfamily that catalyzes the conversion of GDP-L-galactose to L-galactose 1-phosphate in a reaction that consumes inorganic phosphate and produces GDP. In characterizing recombinant VTC2 from A. thaliana as a specific GDP-L-galactose/GDP-D-glucose phosphorylase, we conclude that enzymes catalyzing each of the ten steps of the Smirnoff-Wheeler pathway from glucose to ascorbate have been identified. Finally, we identify VTC2 homologs in plants, invertebrates, and vertebrates, suggesting that a similar reaction is used widely in nature. PMID- 17462989 TI - Transcriptional regulation of Cidea, mitochondrial cell death-inducing DNA fragmentation factor alpha-like effector A, in mouse liver by peroxisome proliferator-activated receptor alpha and gamma. AB - Cidea (cell death-inducing DNA fragmentation factor alpha-like effector A), a member of a novel family of proapoptotic proteins, is expressed abundantly in the brown adipose tissue of the mouse. Although Cidea mRNA is not detectable in the mouse liver, we now show that peroxisome proliferator-activated receptor (PPAR) alpha ligands Wy-14,643 and ciprofibrate increase the Cidea mRNA level in a PPARalpha-dependent manner, whereas Cidea induction in liver by PPARgamma overexpression is PPARalpha independent. Increase in Cidea mRNA content in liver did not alter the expression of uncoupling protein 1 (Ucp1) gene, which regulates thermogenesis, lipolysis, and conservation of energy. Although Cidea is considered to be a proapoptotic factor, Cidea induction in liver did not result in increased apoptosis. To elucidate the mechanism by which PPARalpha and PPARgamma regulate Cidea gene expression in the liver, we analyzed the promoter region of the Cidea gene. Three putative peroxisome proliferator response elements (PPREs) are found in the Cidea gene promoter. Transactivation, gel shift, and chromatin immunoprecipitation assays indicated that the proximal PPRE in Cidea gene (Cidea-PPRE1 at -680/-668) is functional for both PPARalpha and gamma. We conclude that Cidea is a novel target gene for both PPARalpha and gamma in the liver where these two transcription factors utilize the same PPRE region for dual regulation. The induction of Cidea in liver with these PPARalpha and -gamma agonists suggests a possible role for Cidea in energy metabolism and a less likely role in hepatocyte apoptosis. PMID- 17462990 TI - Biochemical and genetic studies of UBR3, a ubiquitin ligase with a function in olfactory and other sensory systems. AB - Our previous work identified E3 ubiquitin ligases, termed UBR1-UBR7, that contain the approximately 70-residue UBR box, a motif important for the targeting of N end rule substrates. In this pathway, specific N-terminal residues of substrates are recognized as degradation signals by UBR box-containing E3s that include UBR1, UBR2, UBR4, and UBR5. The other E3s of this set, UBR3, UBR6, and UBR7, remained uncharacterized. Here we describe the cloning and analyses of mouse UBR3. The similarities of UBR3 to the UBR1 and UBR2 E3s of the N-end rule pathway include the RING and UBR domains. We show that HR6A and HR6B, the E2 enzymes that bind to UBR1 and UBR2, also interact with UBR3. However, in contrast to UBR1 and UBR2, UBR3 does not recognize N-end rule substrates. We also constructed UBR3 lacking mouse strains. In the 129SvImJ background, UBR3-/- mice died during embryogenesis, whereas the C57BL/6 background UBR3-/- mice exhibited neonatal lethality and suckling impairment that could be partially rescued by litter size reduction. The adult UBR3-/- mice had female-specific behavioral anosmia. Cells of the olfactory pathway were found to express beta-galactosidase (LacZ) that marked the deletion/disruption UBR3- allele. The UBR3-specific LacZ expression was also prominent in cells of the touch, vision, hearing, and taste systems, suggesting a regulatory role of UBR3 in sensory pathways, including olfaction. By analogy with functions of the UBR domain in the N-end rule pathway, we propose that the UBR box of UBR3 may recognize small compounds that modulate the targeting, by this E3, of its currently unknown substrates. PMID- 17462991 TI - Distinct uptake mechanisms but similar intracellular processing of two different toll-like receptor ligand-peptide conjugates in dendritic cells. AB - Covalent conjugation of Toll-like receptor ligands (TLR-L) to synthetic antigenic peptides strongly improves antigen presentation in vitro and T lymphocyte priming in vivo. These molecularly well defined TLR-L-peptide conjugates, constitute an attractive vaccination modality, sharing the peptide antigen and a defined adjuvant in one single molecule. We have analyzed the intracellular trafficking and processing of two TLR-L conjugates in dendritic cells (DCs). Long synthetic peptides containing an ovalbumin cytotoxic T-cell epitope were chemically conjugated to two different TLR-Ls the TLR2 ligand, Pam(3)CysSK(4) (Pam) or the TLR9 ligand CpG. Rapid and enhanced uptake of both types of TLR-L-conjugated peptide occurred in DCs. Moreover, TLR-L conjugation greatly enhanced antigen presentation, a process that was dependent on endosomal acidification, proteasomal cleavage, and TAP translocation. The uptake of the CpG approximately conjugate was independent of endosomally-expressed TLR9 as reported previously. Unexpectedly, we found that Pam approximately conjugated peptides were likewise internalized independently of the expression of cell surface-expressed TLR2. Further characterization of the uptake mechanisms revealed that TLR2-L employed a different uptake route than TLR9-L. Inhibition of clathrin- or caveolin-dependent endocytosis greatly reduced uptake and antigen presentation of the Pam-conjugate. In contrast, internalization and antigen presentation of CpG approximately conjugates was independent of clathrin-coated pits but partly dependent on caveolae formation. Importantly, in contrast to the TLR-independent uptake of the conjugates, TLR expression and downstream TLR signaling was required for dendritic cell maturation and for priming of naive CD8(+) T-cells. Together, our data show that targeting to two distinct TLRs requires distinct uptake mechanism but follows similar trafficking and intracellular processing pathways leading to optimal antigen presentation and T-cell priming. PMID- 17462992 TI - Prostaglandin endoperoxide H synthases: peroxidase hydroperoxide specificity and cyclooxygenase activation. AB - The cyclooxygenase (COX) activity of prostaglandin endoperoxide H synthases (PGHSs) converts arachidonic acid and O2 to prostaglandin G2 (PGG2). PGHS peroxidase (POX) activity reduces PGG2 to PGH2. The first step in POX catalysis is formation of an oxyferryl heme radical cation (Compound I), which undergoes intramolecular electron transfer forming Intermediate II having an oxyferryl heme and a Tyr-385 radical required for COX catalysis. PGHS POX catalyzes heterolytic cleavage of primary and secondary hydroperoxides much more readily than H2O2, but the basis for this specificity has been unresolved. Several large amino acids form a hydrophobic "dome" over part of the heme, but when these residues were mutated to alanines there was little effect on Compound I formation from H2O2 or 15-hydroperoxyeicosatetraenoic acid, a surrogate substrate for PGG2. Ab initio calculations of heterolytic bond dissociation energies of the peroxyl groups of small peroxides indicated that they are almost the same. Molecular Dynamics simulations suggest that PGG2 binds the POX site through a peroxyl-iron bond, a hydrogen bond with His-207 and van der Waals interactions involving methylene groups adjoining the carbon bearing the peroxyl group and the protoporphyrin IX. We speculate that these latter interactions, which are not possible with H2O2, are major contributors to PGHS POX specificity. The distal Gln-203 four residues removed from His-207 have been thought to be essential for Compound I formation. However, Q203V PGHS-1 and PGHS-2 mutants catalyzed heterolytic cleavage of peroxides and exhibited native COX activity. PGHSs are homodimers with each monomer having a POX site and COX site. Cross-talk occurs between the COX sites of adjoining monomers. However, no cross-talk between the POX and COX sites of monomers was detected in a PGHS-2 heterodimer comprised of a Q203R monomer having an inactive POX site and a G533A monomer with an inactive COX site. PMID- 17462993 TI - Awaking TIM22, a dynamic ligand-gated channel for protein insertion in the mitochondrial inner membrane. AB - Aqueous channels are at the core of the translocase of the outer membrane (TOM) and the translocase of the inner membrane for the transport of preproteins (TIM23), the translocases mediating the transport of proteins across the outer and inner mitochondrial membranes. Yet, the existence of a channel associated to the translocase of the inner membrane for the insertion of multitopic protein (TIM22) complex has been arguable, as its function relates to the insertion of multispanning proteins into the inner membrane. For the first time, we report conditions for detecting a channel activity associated to the TIM22 translocase in organelle, i.e. intact mitoplasts. An internal signal peptide in the intermembrane space of mitochondria is a requisite to inducing this channel, which is otherwise silent. The channel showed slightly cationic and high conductance activity of 1000 pS with a predominant half-open substate. Despite their different composition, the channels of the three mitochondrial translocases were thus remarkably similar, in agreement with their common task as pores transiently trapping proteins en route to their final destination. The opening of the TIM22 channel was a step-up process depending on the signal peptide concentration. Interestingly, low membrane potentials kept the channel fully open, providing a threshold level of the peptide is present. Our results portray TIM22 as a dynamic channel solely active in the presence of its cargo proteins. In its fully open conformation, favored by the combined action of internal signal peptide and low membrane potential, the channel could embrace the in-transit protein. As insertion progressed and initial interaction with the signal peptide faded, the channel would close, sustaining its role as a shunt that places trapped proteins into the membrane. PMID- 17462994 TI - MicroRNA-124a regulates Foxa2 expression and intracellular signaling in pancreatic beta-cell lines. AB - MicroRNAs (miRNAs) are short non-coding RNAs that have been implicated in fine tuning gene regulation, although the precise roles of many are still unknown. Pancreatic development is characterized by the complex sequential expression of a gamut of transcription factors. We have performed miRNA expression profiling at two key stages of mouse embryonic pancreas development, e14.5 and e18.5. miR 124a2 expression was strikingly increased at e18.5 compared with e14.5, suggesting a possible role in differentiated beta-cells. Among the potential miR 124a gene targets identified by biocomputation, Foxa2 is known to play a role in beta-cell differentiation. To evaluate the impact of miR-124a2 on gene expression, we overexpressed or down-regulated miR-124a2 in MIN6 beta-cells. As predicted, miR-124a2 regulated Foxa2 gene expression, and that of its downstream target, pancreatic duodenum homeobox-1 (Pdx-1). Foxa2 has been described as a master regulator of pancreatic development and also of genes involved in glucose metabolism and insulin secretion, including the ATP-sensitive K(+) (K(ATP)) channel subunits, Kir6.2 and Sur-1. Correspondingly, miR-124a2 overexpression decreased, and anti-miR-124a2 increased Kir6.2 and Sur-1 mRNA levels. Moreover, miR-124a2 modified basal and glucose- or KCl-stimulated intracellular free Ca(2+) concentrations in single MIN6 and INS-1 (832/13) beta-cells, without affecting the secretion of insulin or co-transfected human growth hormone, consistent with an altered sensitivity of the beta-cell exocytotic machinery to Ca(2+). In conclusion, whereas the precise role of microRNA-124a2 in pancreatic development remains to be deciphered, we identify it as a regulator of a key transcriptional protein network in beta-cells responsible for modulating intracellular signaling. PMID- 17462995 TI - Binding of pleomorphic adenoma gene-like 2 to the tumor necrosis factor (TNF) alpha-responsive region of the NCF2 promoter regulates p67(phox) expression and NADPH oxidase activity. AB - NCF2, the gene encoding the NADPH oxidase cytosolic component p67(phox), is up regulated by TNF-alpha, and we recently mapped a region in the NCF2 promoter that was required for this TNF-alpha-dependent response. Because this TNF-alpha responsive region (TRR) lacked recognizable transcription factor binding elements, we performed studies to identify factors involved in regulating NCF2 via the TRR. Using the TRR sequence as bait in a yeast one-hybrid screen, we identified the zinc finger transcription factor Pleomorphic Adenoma Gene-Like 2 (PLAGL2) as a candidate regulator of NCF2 expression. PLAGL2-specific antibodies were generated that detected the native and SUMO1-modified forms of endogenous PLAGL2. EMSA and DNA-binding protein affinity purification analyses demonstrated specific binding of in vitro-translated as well as endogenously expressed PLAGL2 to the TRR, and chromatin immunoprecipitation assays demonstrated enhanced binding of endogenous PLAGL2 to the TRR in vivo with TNF-alpha treatment. Knockdown of PLAGL2 protein inhibited up-regulation of NCF2 transcript, p67(phox) protein expression, and subsequent superoxide production in response to TNF alpha. Furthermore, relative levels of native and SUMO1-modified endogenous PLAGL2 protein were modulated in a time-dependant manner in response to TNF-alpha treatment. These data clearly identify PLAGL2 as a novel regulator of NCF2 gene expression as well as NADPH oxidase activity and contribute to a greater understanding of the transcriptional regulation of NCF2. PMID- 17462996 TI - Caspase-8 and c-FLIPL associate in lipid rafts with NF-kappaB adaptors during T cell activation. AB - Humans and mice lacking functional caspase-8 in T cells manifest a profound immunodeficiency syndrome due to defective T cell antigen receptor (TCR)-induced NF-kappaB signaling and proliferation. It is unknown how caspase-8 is activated following T cell stimulation, and what is the caspase-8 substrate(s) that is necessary to initiate T cell cycling. We observe that following TCR ligation, a small portion of total cellular caspase-8 and c-FLIP(L) rapidly migrate to lipid rafts where they associate in an active caspase complex. Activation of caspase-8 in lipid rafts is followed by rapid cleavage of c-FLIP(L) at a known caspase-8 cleavage site. The active caspase.c-FLIP complex forms in the absence of Fas (CD95/APO1) and associates with the NF-kappaB signaling molecules RIP1, TRAF2, and TRAF6, as well as upstream NF-kappaB regulators PKC theta, CARMA1, Bcl-10, and MALT1, which connect to the TCR. The lack of caspase-8 results in the absence of MALT1 and Bcl-10 in the active caspase complex. Consistent with this observation, inhibition of caspase activity attenuates NF-kappaB activation. The current findings define a link among TCR, caspases, and the NF-kappaB pathway that occurs in a sequestered lipid raft environment in T cells. PMID- 17462997 TI - A systematic and comprehensive combinatorial approach to simultaneously improve the activity, reaction specificity, and thermal stability of p-hydroxybenzoate hydroxylase. AB - We have simultaneously improved the activity, reaction specificity, and thermal stability of p-hydroxybenzoate hydroxylase by means of systematic and comprehensive combinatorial mutagenesis starting from available single mutations. Introduction of random mutations at the positions of four cysteine and eight methionine residues provided 216 single mutants as stably expressed forms in Escherichia coli host cells. Four characteristics, hydroxylase activity toward p hydroxybenzoate (main activity), protocatechuate-dependent NADPH oxidase activity (sub-activity), ratio of sub-activity to main activity (reaction specificity), and thermal stability, of the purified mutants were determined. To improve the above characteristics for diagnostic use of the enzyme, 11 single mutations (C152V, C211I, C332A, M52V, M52Q, M110L, M110I, M213G, M213L, M276Q, and M349A) were selected for further combinatorial mutagenesis. All possible combinations of the mutations provided 18 variants with double mutations and further combinatorial mutagenesis provided 6 variants with triple mutations and 9 variants with quadruple mutations with the simultaneously improved four properties. PMID- 17462998 TI - Myosin Vb is required for trafficking of the cystic fibrosis transmembrane conductance regulator in Rab11a-specific apical recycling endosomes in polarized human airway epithelial cells. AB - Cystic fibrosis transmembrane conductance regulator (CFTR)-mediated Cl(-) secretion across fluid-transporting epithelia is regulated, in part, by modulating the number of CFTR Cl(-) channels in the plasma membrane by adjusting CFTR endocytosis and recycling. However, the mechanisms that regulate CFTR recycling in airway epithelial cells remain unknown, at least in part, because the recycling itineraries of CFTR in these cells are incompletely understood. In a previous study, we demonstrated that CFTR undergoes trafficking in Rab11a specific apical recycling endosomes in human airway epithelial cells. Myosin Vb is a plus-end-directed, actin-based mechanoenzyme that facilitates protein trafficking in Rab11a-specific recycling vesicles in several cell model systems. There are no published studies examining the role of myosin Vb in airway epithelial cells. Thus, the goal of this study was to determine whether myosin Vb facilitates CFTR recycling in polarized human airway epithelial cells. Endogenous CFTR formed a complex with endogenous myosin Vb and Rab11a. Silencing myosin Vb by RNA-mediated interference decreased the expression of wild-type CFTR and DeltaF508-CFTR in the apical membrane and decreased CFTR-mediated Cl(-) secretion across polarized human airway epithelial cells. A recombinant tail domain fragment of myosin Vb attenuated the plasma membrane expression of CFTR by arresting CFTR recycling. The dominant-negative effect was dependent on the ability of the myosin Vb tail fragment to interact with Rab11a. Taken together, these data indicate that myosin Vb is required for CFTR recycling in Rab11a specific apical recycling endosomes in polarized human airway epithelial cells. PMID- 17462999 TI - Homooligomeric and heterooligomeric associations between K+-Cl- cotransporter isoforms and between K+-Cl- and Na+-K+-Cl- cotransporters. AB - Little is known regarding the quaternary structure of cation-Cl- cotransporters (CCCs) except that the Na+-dependent CCCs can exist as homooligomeric units. Given that each of the CCCs exhibits unique functional properties and that several of these carriers coexist in various cell types, it would be of interest to determine whether the four K+-Cl- cotransporter (KCC) isoforms and their splice variants can also assemble into such units and, more importantly, whether they can form heterooligomers by interacting with each other or with the secretory Na+-K+-Cl- cotransporter (NKCC1). In the present work, we have addressed these questions by conducting two groups of analyses: 1) yeast two hybrid and pull-down assays in which CCC-derived protein segments were used as both bait and prey and 2) coimmunoprecipitation and functional studies of intact CCCs coexpressed in Xenopus laevis oocytes. Through a combination of such analyses, we have found that KCC2 and KCC4 could adopt various oligomeric states (in the form of KCC2-KCC2, KCC4-KCC4, KCC2-KCC4, and even KCC4-NKCC1 complexes), that their carboxyl termini were probably involved in carrier assembly, and that the KCC4-NKCC1 oligomers, more specifically, could deploy unique functional features. Through additional coimmunoprecipitation studies, we have also found that KCC1 and KCC3 had the potential of assembling into various types of CCC-CCC oligomers as well, although the interactions uncovered were not characterized as extensively, and the protein segments involved were not identified in yeast two hybrid assays. Taken together, these findings could change our views on how CCCs operate or are regulated in animal cells by suggesting, in particular, that cation-Cl- cotransport achieves higher levels of functional diversity than foreseen. PMID- 17463000 TI - Regulation of GREB1 transcription by estrogen receptor alpha through a multipartite enhancer spread over 20 kb of upstream flanking sequences. AB - Estrogen receptors activate transcription in part through direct interactions with specific DNA motifs, called estrogen response elements (EREs). Here we show that the strong and sustained induction of the gene regulated in breast cancer 1 (GREB1), a gene of unknown function that has been previously suggested to play a role in the effects of estradiol on breast cancer cell proliferation (Rae, J. M., Johnson, M. D., Scheys, J. O., Cordero, K. E., Larios, J. M., and Lippman, M. E. (2005) Breast Cancer Res. Treat 92, 141-149), is mediated by binding of estrogen receptor alpha (ERalpha) to three consensus EREs spread over approximately 20 kb of upstream flanking sequences. In addition to ERalpha, coactivator SRC-3, acetylated histones and phosphorylated RNA polymerase II (P-polII) were detected on all three EREs in the presence of estrogen, while basal recruitment of ERalpha and P-polII was observed only on the proximal element. Chromatin loops were formed between each ERE and the GREB1 transcriptional start site in the presence of estrogen but not of a total antiestrogen. Furthermore, estradiol induced physical association between EREs, suggesting that these elements function as a potent multipartite enhancer to regulate GREB1 transcription. PMID- 17463001 TI - Serine 64 phosphorylation enhances the antiapoptotic function of Mcl-1. AB - Mcl-1 is an antiapoptotic Bcl-2 family member that is highly regulated and when dysregulated contributes to cancer. The Mcl-1 protein is phosphorylated at multiple sites in response to different signaling events. Phosphorylations at Thr163 (by ERK) and Ser159 (by glycogen-synthase kinase 3beta) have recently been shown to slow and enhance, respectively, Mcl-1 protein turnover. Phosphorylation is also known to be stimulated at other, as-yet uncharacterized sites in the G2/M phase of the cell cycle. Using an S peptide-tagged Mcl-1 T163A mutant, Ser64 was identified as a novel Mcl-1 phosphorylation site by mass spectrometry. Immunoblotting demonstrated that phosphorylation at this site was maximal in cells in G2/M phase, was enhanced by tumor necrosis factor-alpha-related apoptosis-inducing ligand (TRAIL) treatment, was blocked by inhibitors of CDK (but not ERK or glycogen-synthase kinase 3beta), and was stimulated in vitro by CDK 1, CDK2, and JNK1. The half-life of a nonphosphorylatable S64A Mcl-1 mutant was indistinguishable from that of the wild type polypeptide. In contrast, this mutant failed to protect cells from TRAIL-mediated apoptosis, whereas reconstitution with the phosphomimetic S64E Mcl-1 mutant rendered cells TRAIL resistant. This anti-apoptotic phenotype of the S64E Mcl-1 mutant was also associated with enhanced binding to the proapoptotic proteins Bim, Noxa, and Bak. A pharmacological CDK inhibitor that reduced Ser64 phosphorylation also sensitized cells to TRAIL cytotoxicity. Collectively, these observations not only identify G2/M-associated phosphorylation at Ser64 as a critical determinant of the antiapoptotic activity of Mcl-1 but also elucidate a novel mechanism by which CDK1/2 inhibitors can enhance the effectiveness of the cytotoxic cytokine TRAIL. PMID- 17463002 TI - Phosphorylation of human CTP synthetase 1 by protein kinase C: identification of Ser(462) and Thr(455) as major sites of phosphorylation. AB - Phosphorylation of human CTP synthetase 1 by mammalian protein kinase C was examined. Using purified Escherichia coli-expressed CTP synthetase 1 as a substrate, protein kinase C activity was time- and dose-dependent and dependent on the concentrations of ATP and CTP synthetase 1. The protein kinase C phosphorylation of the recombinant enzyme was accompanied by a 95-fold increase in CTP synthetase 1 activity. Phosphopeptide mapping and phosphoamino acid analyses showed that CTP synthetase 1 was phosphorylated on multiple serine and threonine residues. The induction of PKC1(R398A)-encoded protein kinase C resulted in a 50% increase for human CTP synthetase 1 phosphorylation in the Saccharomyces cerevisiae ura7Delta ura8Delta mutant lacking yeast CTP synthetase activity. Synthetic peptides that contain the protein kinase C motif for Ser(462) and Thr(455) were substrates for mammalian protein kinase C, and S462A and T455A mutations resulted in decreases in the extent of CTP synthetase 1 phosphorylation that occurred in vivo. Phosphopeptide mapping analysis of S. cerevisiae-expressed CTP synthetase 1 mutant enzymes phosphorylated with mammalian protein kinase C confirmed that Ser(462) and Thr(455) were phosphorylation sites. The S. cerevisiae-expressed and purified S462A mutant enzyme exhibited a 2-fold reduction in CTP synthetase 1 activity, whereas the purified T455A mutant enzyme exhibited a 2-fold elevation in CTP synthetase 1 activity (Choi, M.-G., and Carman, G.M. (2006) J. Biol. Chem. 282, 5367-5377). These data indicated that protein kinase C phosphorylation at Ser(462) stimulates human CTP synthetase 1 activity, whereas phosphorylation at Thr(455) inhibits activity. PMID- 17463003 TI - The crystal structure of the ivy Delta4-16:0-ACP desaturase reveals structural details of the oxidized active site and potential determinants of regioselectivity. AB - The multifunctional acyl-acyl carrier protein (ACP) desaturase from Hedera helix (English ivy) catalyzes the Delta(4) desaturation of 16:0-ACP and the Delta(9) desaturation of 18:0-ACP and further desaturates Delta(9)-16:1 or Delta(9)-18:1 to the corresponding Delta(4,9) dienes. The crystal structure of the enzyme has been solved to 1.95 A resolution, and both the iron-iron distance of approximately 3.2A and the presence of a mu-oxo bridge reveal this to be the only reported structure of a desaturase in the oxidized FeIII-FeIII form. Significant differences are seen between the oxidized active site and the reduced active site of the Ricinus communis (castor) desaturase; His(227) coordination to Fe2 is lost, and the side chain of Glu(224), which bridges the two iron ions in the reduced structure, does not interact with either iron. Although carboxylate shifts have been observed on oxidation of other diiron proteins, this is the first example of the residue moving beyond the coordination range of both iron ions. Comparison of the ivy and castor structures reveal surface amino acids close to the annulus of the substrate-binding cavity and others lining the lower portion of the cavity that are potential determinants of their distinct substrate specificities. We propose a hypothesis that differences in side chain packing explains the apparent paradox that several residues lining the lower portion of the cavity in the ivy desaturase are bulkier than their equivalents in the castor enzyme despite the necessity for the ivy enzyme to accommodate three more carbons beyond the diiron site. PMID- 17463004 TI - Murine ORAI2 splice variants form functional Ca2+ release-activated Ca2+ (CRAC) channels. AB - The stimulation of membrane receptors coupled to the phopholipase C pathway leads to activation of the Ca(2+) release-activated Ca(2+) (CRAC) channels. Recent evidence indicates that ORAI1 is an essential pore subunit of CRAC channels. STIM1 is additionally required for CRAC channel activation. The present study focuses on the genomic organization, tissue expression pattern, and functional properties of the murine ORAI2. Additionally, we report the cloning of the murine ORAI1, ORAI3, and STIM1. Two chromosomal loci were identified for the murine orai2 gene, one containing an intronless gene and a second locus that gives rise to the splice variants ORAI2 long (ORAI2L) and ORAI2 short (ORAI2S). Northern blots revealed a prominent expression of the ORAI2 variants in the brain, lung, spleen, and intestine, while ORAI1, ORAI3, and STIM1 appeared to be near ubiquitously expressed in mice tissues. Specific antibodies detected ORAI2 in RBL 2H3 but not in HEK 293 cells, whereas both cell lines appeared to express ORAI1 and STIM1 proteins. Co-expression experiments with STIM1 and either ORAI1 or ORAI2 variants showed that ORAI2L and ORAI2S enhanced substantially CRAC current densities in HEK 293 but were ineffective in RBL 2H3 cells, whereas ORAI1 strongly amplified CRAC currents in both cell lines. Thus, the capability of ORAI2 variants to form CRAC channels depends strongly on the cell background. Additionally, CRAC channels formed by ORAI2S were strongly sensitive to inactivation by internal Ca(2+). When co-expressed with STIM1 and ORAI1, ORAI2S apparently plays a negative dominant role in the formation of CRAC channels. PMID- 17463005 TI - A six-membrane-spanning topology for yeast and Arabidopsis Tsc13p, the enoyl reductases of the microsomal fatty acid elongating system. AB - The very long chain fatty acids are crucial building blocks of essential lipids, most notably the sphingolipids. These elongated fatty acids are synthesized by a system of enzymes that are organized in a complex within the endoplasmic reticulum membrane. Although several of the components of the elongase complex have recently been identified, little is known about how these proteins are organized within the membrane or about how they interact with one another during fatty acid elongation. In this study the topology of Tsc13p, the enoyl reductase of the elongase system, was investigated. The N and C termini of Tsc13p reside in the cytoplasm, and six putative membrane-spanning domains were identified by insertion of glycosylation and factor Xa cleavage sites at various positions. The N-terminal domain including the first membrane-spanning segment contains sufficient information for targeting to the endoplasmic reticulum membrane. Studies of the Arabidopsis Tsc13p protein revealed a similar topology. Highly conserved domains of the Tsc13p proteins that are likely to be important for enzymatic activity lie on the cytosolic face of the endoplasmic reticulum, possibly partially embedded within the membrane. PMID- 17463007 TI - Septal alcohol ablation in hypertrophic obstructive cardiomyopathy: improving cardiac function by generating a myocardial scar. PMID- 17463008 TI - Seeing is believing: acute haemodynamic response to predict long-term outcome in cardiac resynchronization therapy. PMID- 17463009 TI - Heart failure: are we neglecting the silent majority? PMID- 17463010 TI - Guidelines on the management of valvular heart disease. PMID- 17463011 TI - Association of diarrhea with anemia among children under age five living in rural areas of Indonesia. AB - The high incidence of anemia of infection among children in developing countries is not well characterized. We investigated the relationship between diarrhea, fever and other risk factors for anemia in young children in the community. The relationship between risk factors for anemia was examined in a cross-sectional study of 85 229 children, aged 6-59 months, from impoverished families in rural areas of Indonesia. The prevalence of anemia was 56.1% among the study subjects. Those considered anemic were more likely to be younger, male, stunted, underweight, wasted, to have low maternal and paternal education and to have current diarrhea or history of diarrhea in the previous 7 days compared with children without anemia (all P < 0.0001). In separate multivariate models adjusted for age, sex, stunting, maternal age and education, and weekly per capita household expenditure, current diarrhea (OR 1.15, 95% CI 1.07-1.325, P < 0.0001) and a history of diarrhea in the previous 7 days (OR 1.16, 95% CI 1.09 1.25, P < 0.0001) were associated with an increased risk of anemia. In similar models, current fever had a borderline association with anemia (OR 1.14, 95% CI 0.98-1.32, P = 0.09). We conclude that diarrhea is a contributing factor of anemia among young children living in rural areas in Indonesia. PMID- 17463012 TI - National vitamin A supplementation coverage survey among 6-59 months old children in Guinea (West Africa). AB - BACKGROUND: Vitamin A deficiency (VAD) is a major contributor to child mortality in sub-Saharan Africa. Sustained control of VAD is essential to meet the Millennium Development Goal (MDG) for reduction of child mortality in Guinea. Since, twice yearly vitamin A supplementation (VAS), either coupled with National Immunization Days or stand-alone has been adopted as a key strategy to combat VAD in 6-59 months old children. OBJECTIVES: The objectives of this survey were to describe national VAS coverage rates and related factors affecting VAS coverage among 6-59 months old Guinean children. METHODS: In July 2003, a VAS coverage survey was implemented in Guinea. A cross-sectional random cluster survey was conducted to select Guinean children (n = 1950, 390 mother-child pairs per zone) aged 6-59 months. The country was divided into four agro-ecological zones with 30 clusters chosen per zone. Within each cluster, a random selection of 13 households with at least one child was carried out with random selection of one child per household. Data on characteristics of children, receipt of VAS, caregivers' socio-economic characteristics, vitamin A knowledge and practices of caregivers were collected by questionnaire. RESULTS: The national coverage rate of 68% is much lower than the official coverage rate of 93%. Middle Guinea, the region most affected by VAD, had the lowest coverage rate (58%). CONCLUSION: In order to increase overall VAS coverage and reduce regional disparities, it is suggested that mass VAS be organized on a regional level, prioritizing rural regions (Middle and Upper Guinea) and the city of Conakry. PMID- 17463013 TI - For public service or money: understanding geographical imbalances in the health workforce. AB - Geographical imbalances in the health workforce have been a consistent feature of nearly all health systems, and especially in developing countries. In this paper we investigate the willingness to work in a rural area among final year nursing and medical students in Ethiopia. Analysing data obtained from contingent valuation questions for final year students from three medical schools and eight nursing schools, we find that there is substantial heterogeneity in the willingness to serve in rural areas. Using both ordinary least squares and maximum likelihood regression analysis, we find that household consumption and the student's motivation to help the poor are the main determinants of willingness to work in a rural area. We carry out a simulation on how much it would cost to get a target proportion of health workers to take up a rural post. PMID- 17463014 TI - Reproductive health and midwives: does occupational status differentiate their attitudes on assisted reproduction technologies from those of the general population? AB - BACKGROUND: Advancements within assisted reproduction technologies (ART) raise ethical questions; however, research on health care professionals' attitudes towards their application is limited. This study aimed at assessing certified (CMs) and Student (SMs) midwives' attitudes towards various aspects of ART as well as comparing them with public opinion. METHODS: The final sample included 567 female CMs and 605 women from the general population (age range: 25-62 years), 221 SMs and 209 female non-SMs (age range: 18-24 years). The questionnaire administered included socio-demographic information, items addressing knowledge issues and attitude statements. Data were analysed using principal components analysis, one-way analysis of variance and Friedman's test, as well as multiple linear regression. RESULTS: Four attitudinal factors emerged: 'genetic counselling' (GC), 'application of ART', 'moral dilemmas' and 'socio ethical aspects'; occupational status did not affect attitudes towards GC, however SMs expressed more positive attitudes regarding the latter three factors (P<0.001: 17.49, 14.14 and 11.55). Student groups expressed more negative attitudes for multifetal pregnancy reduction (SMs: 1.88+/-0.83; non-SMs: 2.17+/ 0.77) whereas the other two groups were least favourable towards embryo donation (2.30+/-0.80, CM; 2.32+/-0.83, general population). Sex selection and the use of ART by menopausal or homosexual women were the least acceptable practices for all groups (P<0.001). A high level of relevant knowledge was positively associated with 'application of ART' and acceptability of its use by specific population groups (b=0.469, b=0.19). Findings on factors influencing attitudinal patterns are further discussed. CONCLUSIONS: In this first attempt, it was revealed that CMs express the same conservative attitudes as the general population. PMID- 17463015 TI - A quantitative model for linking two disparate sets of articles in MEDLINE. AB - BACKGROUND: Identifying information that implicitly links two disparate sets of articles is a fundamental and intuitive data mining strategy that can help investigators address real scientific questions. The Arrowsmith two-node search finds title words and phrases (so-called B-terms) that are shared across two sets of articles within MEDLINE and displays them in a manner that facilitates human assessment. A serious stumbling-block has been the lack of a quantitative model for predicting which of the hundreds if not thousands of B-terms computed for a given search are most likely to be relevant to the investigator. METHODOLOGY/PRINCIPAL FINDINGS: Using a public two-node search interface, field testers devised a set of two-node searches under real life conditions and a certain number of B-terms were marked relevant. These were employed as 'gold standards;' each B-term was characterized according to eight complementary features that were strongly correlated with relevance. A logistic regression model was developed that permits one to estimate the probability of relevance for each B-term, to rank B-terms according to their likely relevance, and to estimate the overall number of relevant B-terms inherent in a given two-node search. CONCLUSIONS/SIGNIFICANCE: The model greatly simplifies and streamlines the process of carrying out a two-node search, and may be applicable to a number of other literature-based discovery applications, including the so-called one-node search and related gene-centric strategies that incorporate implicit links to predict how genes may be related to each other and to human diseases. This should encourage much wider exploration of text mining for implicit information among the general scientific community. AVAILABILITY: Two-node searches can be carried out freely at http://arrowsmith.psych.uic.edu. SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online. PMID- 17463016 TI - Faspad: fast signaling pathway detection. AB - Faspad is a user-friendly tool that detects candidates for linear signaling pathways in protein interaction networks based on an approach by Scott et al. (Journal of Computational Biology, 2006). Using recent algorithmic insights, it can solve the underlying NP-hard problem quite fast: for protein networks of typical size (several thousand nodes), pathway candidates of length up to 13 proteins can be found within seconds and with a 99.9% probability of optimality. Faspad graphically displays all candidates that are found; for evaluation and comparison purposes, an overlay of several candidates and the surrounding network context can also be shown. AVAILABILITY: Faspad is available as free software under the GPL license at http://theinf1.informatik.uni-jena.de/faspad/ and runs under Linux and Windows. PMID- 17463017 TI - SciRoKo: a new tool for whole genome microsatellite search and investigation. AB - SciRoKo is a user-friendly software tool for the identification of microsatellites in genomic sequences. The combination of an extremely fast search algorithm with a built-in summary statistic tool makes SciRoKo an excellent tool for full genome analysis. Compared to other already existing tools, SciRoKo also allows the analysis of compound microsatellites. AVAILABILITY: free for use: www.kofler.or.at/Bioinformatics. SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online. PMID- 17463018 TI - The qualitative and time-dependent character of spatial relations in biomedical ontologies. AB - MOTIVATION: The formal representation of mereological aspects of canonical anatomy (parthood relations) is relatively well understood. The formal representation of other aspects of canonical anatomy, such as connectedness and adjacency relations between anatomical parts, their shape and size as well as the spatial arrangement of anatomical parts within larger anatomical structures are, however, much less well understood and represented in existing computational anatomical and bio-medical ontologies only insufficiently. RESULTS: In this article, we provide a methodology of how to incorporate this kind of information into anatomical and bio-medical ontologies by applying techniques of representing qualitative spatial information from Artificial Intelligence. In particular, we focus on how to explicitly take into account the qualitative and time-dependent character of these relations. As a running example, we use the human temporomandibular joint (TMJ). AVAILABILITY: Using the presented methodology, a formal ontology was developed which is accessible on http://www.ifomis.org/bfo/fol. This ontology may help to improve the logical and ontological rigor of bio-medical ontologies such as the OBO relation ontology. PMID- 17463019 TI - SNP detection exploiting multiple sources of redundancy in large EST collections improves validation rates. AB - MOTIVATION: Single nucleotide polymorphism (SNP) detection exploiting redundancy in expressed sequence tag (EST) collections that arises from the presence of transcripts of the same gene from different individuals has been used to generate large collections of SNPs for many species. A second source of redundancy, namely that EST collections can contain multiple transcripts of the same gene from the same individual, can be exploited to distinguish true SNPs from sequencing error. In this article, we demonstrate with Atlantic salmon and pig EST collections that splitting the EST collection in two, detecting SNPs in both subsets, then accepting only cross-validated SNPs increases validation rates. RESULTS: In the pig data set, 676 cross-validated putative SNPs were detected in a collection of 160,689 ESTs. When validating a subset of these by genotyping on MassARRAY 85.1% of SNPs were polymorphic in successful assays. In the salmon data set, 856 cross validated putative SNPs were detected in a collection of 243,674 ESTs. Validation by genotyping showed that 81.0% of the cross-validated putative SNPs were polymorphic in successful assays. AVAILABILITY: Cross-validated SNPs are available at dbSNP (http://www.ncbi.nlm.nih.gov/projects/SNP/), ss69371838 ss69372575 for the salmon SNPs and ss69372587-ss69373226 for the pig SNPs. PMID- 17463020 TI - OSLay: optimal syntenic layout of unfinished assemblies. AB - The whole genome shotgun approach to genome sequencing results in a collection of contigs that must be ordered and oriented to facilitate efficient gap closure. We present a new tool OSLay that uses synteny between matching sequences in a target assembly and a reference assembly to layout the contigs (or scaffolds) in the target assembly. The underlying algorithm is based on maximum weight matching. The tool provides an interactive visualization of the computed layout and the result can be imported into the assembly editing tool Consed to support the design of primer pairs for gap closure. MOTIVATION: To enhance efficiency in the gap closure phase of a genome project it is crucial to know which contigs are adjacent in the target genome. Related genome sequences can be used to layout contigs in an assembly. AVAILABILITY: OSLay is freely available from: http://www ab.informatik.unituebingen.de/software/oslay. PMID- 17463021 TI - Time-varying modeling of gene expression regulatory networks using the wavelet dynamic vector autoregressive method. AB - MOTIVATION: A variety of biological cellular processes are achieved through a variety of extracellular regulators, signal transduction, protein-protein interactions and differential gene expression. Understanding of the mechanisms underlying these processes requires detailed molecular description of the protein and gene networks involved. To better understand these molecular networks, we propose a statistical method to estimate time-varying gene regulatory networks from time series microarray data. One well known problem when inferring connectivity in gene regulatory networks is the fact that the relationships found constitute correlations that do not allow inferring causation, for which, a priori biological knowledge is required. Moreover, it is also necessary to know the time period at which this causation occurs. Here, we present the Dynamic Vector Autoregressive model as a solution to these problems. RESULTS: We have applied the Dynamic Vector Autoregressive model to estimate time-varying gene regulatory networks based on gene expression profiles obtained from microarray experiments. The network is determined entirely based on gene expression profiles data, without any prior biological knowledge. Through construction of three gene regulatory networks (of p53, NF-kappaB and c-myc) for HeLa cells, we were able to predict the connectivity, Granger-causality and dynamics of the information flow in these networks. SUPPLEMENTARY INFORMATION: Additional figures may be found at http://mariwork.iq.usp.br/dvar/. PMID- 17463022 TI - A network-based method for target selection in metabolic networks. AB - MOTIVATION: The lack of new antimicrobials, combined with increasing microbial resistance to old ones, poses a serious threat to public health. With hundreds of genomes sequenced, systems biology promises to help in solving this problem by uncovering new drug targets. RESULTS: Here, we propose an approach that is based on the mapping of the interactions between biochemical agents, such as proteins and metabolites, onto complex networks. We report that nodes and links in complex biochemical networks can be grouped into a small number of classes, based on their role in connecting different functional modules. Specifically, for metabolic networks, in which nodes represent metabolites and links represent enzymes, we demonstrate that some enzyme classes are more likely to be essential, some are more likely to be species-specific and some are likely to be both essential and specific. Our network-based enzyme classification scheme is thus a promising tool for the identification of drug targets. SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online. PMID- 17463023 TI - Meta-analysis of gene expression data: a predictor-based approach. AB - MOTIVATION: With the increasing availability of cancer microarray data sets there is a growing need for integrative computational methods that evaluate multiple independent microarray data sets investigating a common theme or disorder. Meta analysis techniques are designed to overcome the low sample size typical to microarray experiments and yield more valid and informative results than each experiment separately. RESULTS: We propose a new meta-analysis technique that aims at finding a set of classifying genes, whose expression level may be used to answering the classification question in hand. Specifically, we apply our method to two independent lung cancer microarray data sets and identify a joint core subset of genes which putatively play an important role in tumor genesis of the lung. The robustness of the identified joint core set is demonstrated on a third unseen lung cancer data set, where it leads to successful classification using very few top-ranked genes. Identifying such a set of genes is of significant importance when searching for biologically meaningful biomarkers. SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online. PMID- 17463024 TI - An efficient method for the detection and elimination of systematic error in high throughput screening. AB - MOTIVATION: High-throughput screening (HTS) is an early-stage process in drug discovery which allows thousands of chemical compounds to be tested in a single study. We report a method for correcting HTS data prior to the hit selection process (i.e. selection of active compounds). The proposed correction minimizes the impact of systematic errors which may affect the hit selection in HTS. The introduced method, called a well correction, proceeds by correcting the distribution of measurements within wells of a given HTS assay. We use simulated and experimental data to illustrate the advantages of the new method compared to other widely-used methods of data correction and hit selection in HTS. SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online. PMID- 17463025 TI - Group SCAD regression analysis for microarray time course gene expression data. AB - MOTIVATION: Since many important biological systems or processes are dynamic systems, it is important to study the gene expression patterns over time in a genomic scale in order to capture the dynamic behavior of gene expression. Microarray technologies have made it possible to measure the gene expression levels of essentially all the genes during a given biological process. In order to determine the transcriptional factors (TFs) involved in gene regulation during a given biological process, we propose to develop a functional response model with varying coefficients in order to model the transcriptional effects on gene expression levels and to develop a group smoothly clipped absolute deviation (SCAD) regression procedure for selecting the TFs with varying coefficients that are involved in gene regulation during a biological process. RESULTS: Simulation studies indicated that such a procedure is quite effective in selecting the relevant variables with time-varying coefficients and in estimating the coefficients. Application to the yeast cell cycle microarray time course gene expression data set identified 19 of the 21 known TFs related to the cell cycle process. In addition, we have identified another 52 TFs that also have periodic transcriptional effects on gene expression during the cell cycle process. Compared to simple linear regression (SLR) analysis at each time point, our procedure identified more known cell cycle related TFs. CONCLUSIONS: The proposed group SCAD regression procedure is very effective for identifying variables with time-varying coefficients, in particular, for identifying the TFs that are related to gene expression over time. By identifying the TFs that are related to gene expression variations over time, the procedure can potentially provide more insight into the gene regulatory networks. PMID- 17463026 TI - iHMMune-align: hidden Markov model-based alignment and identification of germline genes in rearranged immunoglobulin gene sequences. AB - MOTIVATION: Immunoglobulin heavy chain (IGH) genes in mature B lymphocytes are the result of recombination of IGHV, IGHD and IGHJ germline genes, followed by somatic mutation. The correct identification of the germline genes that make up a variable VH domain is essential to our understanding of the process of antibody diversity generation as well as to clinical investigations of some leukaemias and lymphomas. RESULTS: We have developed iHMMune-align, an alignment program that uses a hidden Markov model (HMM) to model the processes involved in human IGH gene rearrangement and maturation. The performance of iHMMune-align was compared to that of other immunoglobulin gene alignment utilities using both clonally related and randomly selected IGH sequences. This evaluation suggests that iHMMune-align provides a more accurate identification of component germline genes than other currently available IGH gene characterization programs. AVAILABILITY: iHMMune-align cross-platform Java executable and web interface are freely available to academic users and can be accessed at http://www.emi.unsw.edu.au/~ihmmune/. PMID- 17463027 TI - An approximation method for solving the steady-state probability distribution of probabilistic Boolean networks. AB - MOTIVATION: Probabilistic Boolean networks (PBNs) have been proposed to model genetic regulatory interactions. The steady-state probability distribution of a PBN gives important information about the captured genetic network. The computation of the steady-state probability distribution usually includes construction of the transition probability matrix and computation of the steady state probability distribution. The size of the transition probability matrix is 2(n)-by-2(n) where n is the number of genes in the genetic network. Therefore, the computational costs of these two steps are very expensive and it is essential to develop a fast approximation method. RESULTS: In this article, we propose an approximation method for computing the steady-state probability distribution of a PBN based on neglecting some Boolean networks (BNs) with very small probabilities during the construction of the transition probability matrix. An error analysis of this approximation method is given and theoretical result on the distribution of BNs in a PBN with at most two Boolean functions for one gene is also presented. These give a foundation and support for the approximation method. Numerical experiments based on a genetic network are given to demonstrate the efficiency of the proposed method. PMID- 17463028 TI - Identifying the biologically relevant gene categories based on gene expression and biological data: an example on prostate cancer. AB - MOTIVATION: Most gene-expression based studies aim to identify genes with the capability of distinguishing different phenotypes. Although analysis at the genomic level is important, results of the molecular/cellular level are essential for understanding biological mechanisms. To deliver molecular/cellular-level results, a two-stage scheme is widely employed. This scheme just evaluates biological processes/molecular activities individually, totally overlooking the relationship between processes/activities. This treatment conflicts with the fact that most biological processes/molecular activities do not work alone. In order to deliver improved results, this shortcoming should be addressed. RESULTS: We design a selection model from a novel perspective to directly detect important gene functional categories (each category represents a cellular process or a molecular activity). More importantly, the correlations between gene categories are considered. Contributed by this capability, the proposed method shows its advantages over others. AVAILABILITY: the source code in Matlab is accessible via http://www.ee.cityu.edu.hk/~twschow/category_selection/category_selection.htm PMID- 17463029 TI - Two-stage designs applying methods differing in costs. AB - MOTIVATION: Two-stage pilot and integrated designs are powerful tools for investigating large numbers of hypotheses. Asymptotically, optimal two-stage designs controlling the familywise error or false discovery rate are considered when costs and effect sizes per measurement differ between stages and total costs are constrained. RESULTS: Depending on the cost and effect size ratios between the measurements, it is generally more powerful to apply two-stage procedures using one measurement method at both stages. For the practically relevant case that the same method is applied at both stages but designing the second-stage measurements raises extra costs, two-stage designs are more powerful than the single-stage design even for large costs ratios. The power of the optimal pilot and integrated two-stage designs generally are similar, however, the integrated approach is less sensitive even to severe design misspecifications in the planning phase. AVAILABILITY: R-programs (R, 2005) to calculate asymptotically optimal designs are available on: http://statistics.msi.meduniwien.ac.at/index.php?page=ao2stage PMID- 17463030 TI - DITOP: drug-induced toxicity related protein database. AB - MOTIVATION: Drug-induced toxicity related proteins (DITRPs) are proteins that mediate adverse drug reactions (ADRs) or toxicities through their binding to drugs or reactive metabolites. Collection of these proteins facilitates better understanding of the molecular mechanisms of drug-induced toxicity and the rational drug discovery. Drug-induced toxicity related protein database (DITOP) is such a database that is intending to provide comprehensive information of DITRPs. Currently, DITOP contains 1501 records, covering 618 distinct literature reported DITRPs, 529 drugs/ligands and 418 distinct toxicity terms. These proteins were confirmed experimentally to interact with drugs or their reactive metabolites, thus directly or indirectly cause adverse effects or toxicities. Five major types of drug-induced toxicities or ADRs are included in DITOP, which are the idiosyncratic adverse drug reactions, the dose-dependent toxicities, the drug-drug interactions, the immune-mediated adverse drug effects (IMADEs) and the toxicities caused by genetic susceptibility. Molecular mechanisms underlying the toxicity and cross-links to related resources are also provided while available. Moreover, a series of user-friendly interfaces were designed for flexible retrieval of DITRPs-related information. The DITOP can be accessed freely at http://bioinf.xmu.edu.cn/databases/ADR/index.html. SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online. PMID- 17463031 TI - Human recombination rates are increased around accelerated conserved regions- evidence for continued selection? AB - MOTIVATION: We hypothesized that recombination rates might be increased at genetic loci that are subject to more intense selection. Here, we test this hypothesis by using a recently published set of accelerated conserved regions and fine-scale recombination rate estimates provided by the HapMap project. RESULTS: We observed that fine-scale recombination rates are increased around conserved noncoding regions that show accelerated evolution in human or chimp, as compared to noncoding regions showing accelerated evolution in mouse and those being conserved between human and fugu. Recombination rates around hominid accelerated conserved regions (ACRs) are furthermore increased as compared to exonic regions. On the other hand, GC-content is reduced around ACRs, excluding a major confounding influence of GC-content on the observed variation in recombination rate. CONCLUSION: Our observations indicate that selection intensity could be an important determinant of local recombination rate variation and that continued positive selection might act at many ACR loci. Alternatively, a confounding factor needs to be found that causes a congruent signal in recombination rate estimates based on human polymorphism data and in the comparative genomic data. Researchers who consider the explanation involving selection as more likely may expect more common functional sequence variants at ACRs in genetic association studies. PMID- 17463032 TI - Libaffy: software for processing Affymetrix GeneChip data. AB - SUMMARY: Affymetrix GeneChip microarrays are increasingly used in gene expression studies and in greater number. A software library was developed that supports Affymetrix file formats and implements two popular summary algorithms (MAS5.0 and RMA). The library is modular in design for integration into larger systems and processing pipelines. Additionally, a graphical interface (GENE) was developed to allow end-user access to the functionality within the library. AVAILABILITY: libaffy is free to use under the GNU GPL license. The source code and Windows binaries can be freely accessed from the website http://src.moffitt.usf.edu/libaffy. Additional API documentation and user manual are available. PMID- 17463033 TI - A light switch controlling Ca2+-permeable AMPA receptors in the retina. PMID- 17463034 TI - Exciting times in sensory transduction from A(drian) to Z. PMID- 17463035 TI - Heterogeneity of firing properties among rat thalamic reticular nucleus neurons. AB - The thalamic reticular nucleus (TRN) provides inhibitory innervation to most thalamic relay nuclei and receives excitatory innervation from both cortical and thalamic neurons. Ultimately, information transfer through the thalamus to the neocortex is strongly influenced by TRN. In addition, the reciprocal synaptic connectivity between TRN with associated thalamic relay nuclei is critical in generating intrathalamic rhythmic activities that occur during certain arousal states and pathophysiological conditions. Despite evidence suggesting morphological heterogeneity amongst TRN neurons, the heterogeneity of intrinsic properties of TRN neurons has not been systematically examined. One key characteristic of virtually all thalamic neurons is the ability to produce action potentials in two distinct modes: burst and tonic. In this study, we have examined the prevalence of burst discharge within TRN neurons. Our intracellular recordings revealed that TRN neurons can be differentiated by their action potential discharge modes. The majority of neurons in the dorsal TRN (56%) lack burst discharge, and the remaining neurons (35%) show an atypical burst that consists of an initial action potential followed by small amplitude, long duration depolarizations. In contrast, most neurons in ventral TRN (82%) display a stereotypical burst discharge consisting of a transient, high frequency discharge of multiple action potentials. TRN neurons that lack burst discharge typically did not produce low threshold calcium spikes or produced a significantly reduced transient depolarization. Our findings clearly indicate that TRN neurons can be differentiated by differences in their spike discharge properties and these subtypes are not uniformly distributed within TRN. The functional consequences of such intrinsic differences may play an important role in modality-specific thalamocortical information transfer as well as overall circuit level activities. PMID- 17463037 TI - TrkB is necessary for pruning at the climbing fibre-Purkinje cell synapse in the developing murine cerebellum. AB - TrkB, the cognate receptor for brain-derived neurotrophic factor and neurotrophin 4, has been implicated in regulating synapse formation in the central nervous system. Here we asked whether TrkB plays a role in the maturation of the climbing fibre-Purkinje cell (CF-PC) synapse. In rodent cerebellum, Purkinje cells are initially innervated by multiple climbing fibres that are subsequently culled to assume the mature mono-innervated state, and whose contacts translocate from the soma to the dendrites. By employing transgenic mice hypomorphic or null for TrkB expression, our results indicated that perturbation of TrkB in the immature cerebellum resulted in ataxia, that Purkinje cells remained multiply innervated by climbing fibres beyond the normal developmental time frame, and that synaptic transmission at the parallel fibre-Purkinje cell synapse remained functionally unaltered. Mechanistically, we present evidence that attributes the persistence of multiple climbing fibre innervation to an obscured discrimination of relative strengths among competing climbing fibres. Soma-to-dendrite translocation of climbing fibre terminals was unaffected. Thus, TrkB regulates pruning but not translocation of nascent CF-PC synaptic contacts. PMID- 17463036 TI - Neural regulation of rhythmic arm and leg movement is conserved across human locomotor tasks. AB - It has been proposed that different forms of rhythmic human limb movement have a common central neural control ('common core hypothesis'), just as in other animals. We compared the modulation patterns of background EMG and cutaneous reflexes during walking, arm and leg cycling, and arm-assisted recumbent stepping. We hypothesized that patterns of EMG and reflex modulation during cycling and stepping (deduced from mathematical principal components analysis) would be comparable to those during walking because they rely on similar neural substrates. Differences between the tasks were assessed by evoking cutaneous reflexes via stimulation of nerves in the foot and hand in separate trials. The EMG was recorded from flexor and extensor muscles of the arms and legs. Angular positions of the hip, knee and elbow joints were also recorded. Factor analysis revealed that across the three tasks, four principal components explained more than 93% of the variance in the background EMG and middle-latency reflex amplitude. Phase modulation of reflex amplitude was observed in most muscles across all tasks, suggesting activity in similar control networks. Significant correlations between EMG level and reflex amplitude were frequently observed only during static voluntary muscle activation and not during rhythmic movement. Results from a control experiment showed that strong correlation between EMG and reflex amplitudes was observed during discrete, voluntary leg extension but not during walking. There were task-dependent differences in reflex modulation between the three tasks which probably arise owing to specific constraints during each task. Overall, the results show strong correlation across tasks and support common neural patterning as the regulator of arm and leg movement during various rhythmic human movements. PMID- 17463038 TI - Three distinct muscarinic signalling pathways for cationic channel activation in mouse gut smooth muscle cells. AB - Using mutant mice genetically lacking certain subtypes of muscarinic receptor, we have studied muscarinic signal pathways mediating cationic channel activation in intestinal smooth muscle cells. In cells from M2 subtype-knockout (M2-KO) or M3 KO mice, carbachol (100 microM) evoked a muscarinic cationic current (mI(Cat)) as small as approximately 10% of mI(Cat) in wild-type (WT) cells. No appreciable current was evoked in M2/M3 double-KO cells. All mutant type cells preserved normal G-protein-cationic channel coupling. The M3-KO and WT mI(Cat) each showed a U-shaped current-voltage (I-V) relationship, whereas the M2-KO mI(Cat) displayed a linear I-V relationship. Channel analysis in outside-out patches recognized 70-pS and 120-pS channels as the major muscarinic cationic channels. Active patches of M2-KO cells exhibited both 70-pS and 120-pS channel activity usually together, either of which consisted of brief openings (the respective mean open times O(tau) = 0.55 and 0.23 ms). In contrast, active M3-KO patches showed only 70-pS channel activity, which had three open states (O(tau) = 0.55, 3.1 and 17.4 ms). In WT patches, besides the M2-KO and M3-KO types, another type of channel activity was also observed that consisted of 70-pS channel openings with four open states (O(tau) = 0.62, 2.7, 16.9 and 121.1 ms), and patch current of this channel activity showed a U-shaped I-V curve similar to the WT mI(Cat). The present results demonstrate that intestinal myocytes are endowed with three distinct muscarinic pathways mediating cationic channel activation and that the M2/M3 pathway targeting 70-pS channels, serves as the major contributor to mI(Cat) generation. The delineation of this pathway is consistent with the formation of a functional unit by the M2-Go protein and the M3-PLC systems predicted to control cationic channels. PMID- 17463039 TI - PPARdelta expression is influenced by muscle activity and induces slow muscle properties in adult rat muscles after somatic gene transfer. AB - The effects of exercise on skeletal muscle are mediated by a coupling between muscle electrical activity and gene expression. Several activity correlates, such as intracellular Ca(2+), hypoxia and metabolites like free fatty acids (FFAs), might initiate signalling pathways regulating fibre-type-specific genes. FFAs can be sensed by lipid-dependent transcription factors of the peroxisome proliferator activated receptor (PPAR) family. We found that the mRNA for the predominant muscle isoform, PPARdelta, was three-fold higher in the slow/oxidative soleus compared to the fast/glycolytic extensor digitorum longus (EDL) muscle. In histological sections of the soleus, the most oxidative fibres display the highest levels of PPARdelta protein. When the soleus muscle was stimulated electrically by a pattern mimicking fast/glycolytic IIb motor units, the mRNA level of PPARdelta was reduced to less than half within 24 h. In the EDL, a three fold increase was observed after slow type I-like electrical stimulation. When a constitutively active form of PPARdelta was overexpressed for 14 days in normally active adult fibres after somatic gene transfer, the number of I/IIa hybrids in the EDL more than tripled, IIa fibres increased from 14% to 25%, and IIb fibres decreased from 55% to 45%. The level of succinate dehydrogenase activity increased and size decreased, also when compared to normal fibres of the same type. Thus PPARdelta can change myosin heavy chain, oxidative enzymes and size locally in muscle cells in the absence of general exercise. Previous studies on PPARdelta in muscle have been performed in transgenic animals where the transgene has been present during muscle development. Our data suggest that PPARdelta can mediate activity effects acutely in pre-existing adult fibres, and thus is an important link in excitation-transcription coupling. PMID- 17463040 TI - Persistent sodium current in preBotzinger neurons is not necessary for respiratory rhythm generation in the neonatal mouse? PMID- 17463041 TI - On the physiological roles of PIP(2) at cardiac Na+ Ca2+ exchangers and K(ATP) channels: a long journey from membrane biophysics into cell biology. AB - Over the last 10 years we have tried to understand the roles of PIP(2) in regulating cardiac Na(+)-Ca(2+) exchangers and K(ATP) K(+) channels, both of which are directly activated by PIP(2). Up to now, the idea that hormones might physiologically regulate these mechanisms by causing changes of PIP(2) concentrations in the cardiac sarcolemma, either locally or globally, is not well supported. In intact myocardium, but not excised patches, phosphatidylinositol 4 phosphate 5-kinase (PIP5K) activity appears to be Ca(2+) activated and dependent on cardiac activity. Potentially therefore the primary second messenger of the heart, cytoplasmic Ca(2+), may regulate PIP(2) and therewith numerous cardiac membrane processes. In general, however, PIP(2) may simply serve to strongly activate various cardiac channels and transporters when they are inserted in the sarcolemma, while a lack of PIP(2) on internal membranes maintains transporters and channels inactive during trafficking and processing. As in most, if not all, strong regulatory systems of cells, the activating effects of PIP(2) can apparently be countered by strong inactivation mechanisms. In this context, our recent work suggests that internalization of cardiac Na(+)-Ca(2+) exchangers is promoted by increased PIP(2) synthesis, especially in combination with other cell signals. Assuming that multiple adapter-PIP(2) interactions are necessary to initiate the budding of individual membrane vesicles, the dependence of endocytosis on PIP(2) in the surface membrane can potentially be a very steep function. Thus, a better understanding of the regulation of cardiac lipid kinases may be key to understanding when and how cardiac ion transporters and channels are internalized. PMID- 17463042 TI - Presynaptic inhibition differentially shapes transmission in distinct circuits in the mouse retina. AB - Diverse retinal outputs are mediated by ganglion cells that receive excitatory input from distinct classes of bipolar cells (BCs). These classes of BCs separate visual signals into rod, ON and OFF cone pathways. Although BC signalling is a major determinant of the ganglion cell-mediated retinal output, it is not fully understood how light-evoked, presynaptic inhibition from amacrine cell inputs shapes BC outputs. To determine whether differences in presynaptic inhibition uniquely modulate BC synaptic output to specific ganglion cells, we assessed the inhibitory contributions of GABA(A), GABA(C) and glycine receptors across the BC pathways. Here we show that different proportions of GABA(A) and GABA(C) receptor mediated inhibition determined the kinetics of GABAergic presynaptic inhibition across different BC classes. Large, slow GABA(C) and small, fast GABA(A) receptor mediated inputs to rod BCs prolonged light-evoked inhibitory postsynaptic currents (L-IPSCs), while smaller GABA(C) and larger GABA(A) receptor-mediated contributions produced briefer L-IPSCs in ON and OFF cone BCs. Glycinergic inhibition also varied across BC class. In the rod-dominant conditions studied here, slow glycinergic inputs dominated L-IPSCs in OFF cone BCs, attributable to inputs from the rod pathway via AII amacrine cells, while rod and ON cone BCs received little and no glycinergic input, respectively. As these large glycinergic inputs come from rod signalling pathways, in cone-dominant conditions L-IPSCs in OFF cone bipolar cells will probably be dominated by GABA(A) receptor mediated input. Thus, unique presynaptic receptor combinations mediate distinct forms of inhibition to selectively modulate BC outputs, enhancing the distinctions among parallel retinal signals. PMID- 17463043 TI - Selective action of noradrenaline and serotonin on neurones of the spinal superficial dorsal horn in the rat. AB - The superficial dorsal horn of the spinal cord (SDH; laminae I and II) receives strong input from thin primary afferent fibres and is involved in nociception, pain, temperature sensing and other experiences. The SDH also is the target of serotonergic and adrenergic projections from the brain stem. The interaction between descending pathways that utilize particular mediators and the neurone population of the SDH is poorly understood. To explore this issue, in rat spinal cord slices during whole-cell recordings from identified SDH neurones, noradrenaline (NA) or serotonin (5HT) were briefly applied in the superfusing artificial cerebrospinal fluid. The action of these agents proved specifically related to the type of SDH neurone and its dorsal-root afferent input. Vertical, radial and tonic central lamina II cells consistently expressed outward current to both NA and 5HT, but transient central and Substance P (SP)-insensitive lamina I cells were unaffected directly by either NA or 5HT. Extended islet cells responded with outward current to NA and inward current to 5HT. Lamina I SP sensitive cells expressed an outward current regularly to NA. 5HT had inhibitory effects on Adelta and C fibre input to all types of SDH neurones. NA inhibited C fibre input to transient central neurones. The present results support the idea that descending systems may have multiple functions, including but not limited to nociceptive modulation. PMID- 17463044 TI - Ageing and leg postjunctional alpha-adrenergic vasoconstrictor responsiveness in healthy men. AB - Muscle sympathetic vasoconstrictor nerve activity increases with advancing age, but does not result in elevated forearm vasoconstrictor tone because of a selective reduction in alpha1-adrenoceptor responsiveness. In contrast, the leg circulation of older adults is under greater tonic sympathetic vasoconstriction, but it is unclear whether alpha-adrenoceptor responsiveness is altered with age. In the present study, we tested the hypothesis that postjunctional alpha adrenergic vasoconstrictor responsiveness is reduced in the leg circulation with age. We measured femoral blood flow (Doppler ultrasound) and calculated the femoral vascular conductance (FVC) responses to alpha-adrenoceptor stimulation during local blockade of beta-adrenoceptors in 12 young (24 +/- 1 year) and seven healthy older men (62 +/- 2 year). Whole-leg vasoconstrictor responses to local intrafemoral artery infusions of tyramine (evokes noradrenaline (NA) release), phenylephrine (alpha1-agonist) and dexmedetomidine (alpha2-agonist) were assessed. Consistent with previous data, resting femoral blood flow and FVC were approximately 30% lower in older compared with young men (P < 0.05). Maximal vasoconstrictor responses to tyramine (-30 +/- 3 versus -41 +/- 3%), phenylephrine (-25 +/- 4 versus -45 +/- 5%), and dexmedetomidine (-22 +/- 4 versus -44 +/- 3%) were all significantly lower in older compared with young men (all P < 0.05). Our results indicate that human ageing is associated with a reduction in leg postjunctional alpha-adrenoceptor responsiveness to endogenous NA release, and this reduction is evident for both alpha1- and alpha2 adrenoceptors. However, given that basal leg vascular conductance is reduced with age and is primarily mediated by sympathetic vasoconstriction, impaired alpha adrenoceptor responsiveness does not negate the ability of the sympathetic nervous system to evoke greater tonic vasoconstriction in the leg vasculature of older men. PMID- 17463045 TI - Effective release rates at single rat Schaffer collateral-CA1 synapses during sustained theta-burst activity revealed by optical imaging. AB - To understand how information is coded at single hippocampal synapses during high frequency activity, we imaged NMDA receptor-mediated Ca(2+) responses in spines of CA1 neurons using two-photon microscopy. Although discrete quantal events were not readily apparent during continuous theta-burst stimulation (TBS), we found that the steady-state dendritic Ca(2+) response was spatially restricted (half width < 1 microm), voltage dependent and sensitive to MK-801, indicating that that it was mediated by activation of NMDA receptors at single synapses. Partial antagonism of NMDA receptors caused a similar reduction of NMDA EPSCs (measured at the soma) and local dendritic Ca(2+) signals, suggesting that, like EPSCs, the steady-state Ca(2+) signal was made up of a linear addition of quantal events. Statistical analyses of the steady-response suggested that the quantal size did not change dramatically during TBS. Deconvolution of TBS-evoked Ca(2+) responses revealed a heterogeneous population of synapses differing in their capacity to signal high-frequency information, with an average effective steady-state release rate of approximately 2.6 vesicles synapse(-1)s(-1). To assess how the optically determined release rates compare with population measures we analysed the rate of decay of peak EPSCs during train stimulation. From these studies, we estimated a unitary vesicular replenishment rate of 0.02 s(-1), which corresponds to an average release rate of approximately 0.8-2 vesicles s(-1) at individual synapses. Additionally, extracellular recordings from single Schaffer collaterals revealed that spikes propagate reliably during TBS. Hence, during high-frequency activity, Schaffer collaterals conduct spikes with high fidelity, but release quanta with relatively lower efficiency, leaving NMDA receptor function largely intact and synapse specific. Heterogeneity in release rates between synapses suggests that similar patterns of presynaptic action potentials could trigger different forms of plasticity at individual synapses. PMID- 17463046 TI - Mammalian target of rapamycin in the human placenta regulates leucine transport and is down-regulated in restricted fetal growth. AB - Pathological fetal growth is associated with perinatal morbidity and the development of diabetes and cardiovascular disease later in life. Placental nutrient transport is a primary determinant of fetal growth. In human intrauterine growth restriction (IUGR) the activity of key placental amino acid transporters, such as systems A and L, is decreased. However the mechanisms regulating placental nutrient transporters are poorly understood. We tested the hypothesis that the mammalian target of rapamycin (mTOR) signalling pathway regulates amino acid transport in the human placenta and that the activity of the placental mTOR pathway is reduced in IUGR. Using immunohistochemistry and culture of trophoblast cells, we show for the first time that the mTOR protein is expressed in the transporting epithelium of the human placenta. We further demonstrate that placental mTOR regulates activity of the l-amino acid transporter, but not system A or taurine transporters, by determining the mediated uptake of isotope-labelled leucine, methylaminoisobutyric acid and taurine in primary villous fragments after inhibition of mTOR using rapamycin. The protein expression of placental phospho-S6K1 (Thr-389), a measure of the activity of the mTOR signalling pathway, was markedly reduced in placentas obtained from pregnancies complicated by IUGR. These data identify mTOR as an important regulator of placental amino acid transport, and provide a mechanism for the changes in placental leucine transport in IUGR previously demonstrated in humans. We propose that mTOR functions as a placental nutrient sensor, matching fetal growth with maternal nutrient availability by regulating placental nutrient transport. PMID- 17463047 TI - Vitamin B1 (thiamine) uptake by human retinal pigment epithelial (ARPE-19) cells: mechanism and regulation. AB - Retinal abnormality and visual disturbances occur in thiamine-responsive megaloblastic anaemia (TRMA), an autosomal recessive disorder caused by mutations in the human thiamine transporter-1 (hTHTR-1). Human retinal pigment epithelial cells play a pivotal role in supplying thiamine to the highly metabolically active retina but nothing is known about the mechanism, regulation or biological processes involved in thiamine transport in these cells. To address these issues, we used human-derived retinal pigment epithelial ARPE-19 cells to characterize the thiamine uptake process. Thiamine uptake is energy- and temperature dependent, pH-sensitive, Na+-independent, saturable at both the nanomolar (apparent Km, 30 +/- 5 nM) and the micromolar (apparent Km, 1.72 +/- 0.3 microM) concentration ranges, specific for thiamine and sensitive to sulfhydryl group inhibition. The diuretic amiloride caused a concentration-dependent inhibition in thiamine uptake, whereas the anti-trypanosomal drug, melarsoprol, failed to affect the uptake process. Both hTHTR-1 and hTHTR-2 are expressed in ARPE-19 cells as well as in native human retinal tissue with expression of the former being significantly higher than that of the latter. Uptake of thiamine was adaptively regulated by extracellular substrate level via transcriptionally mediated mechanisms that involve both hTHTR-1 and hTHTR-2; it was also regulated by an intracellular Ca2+-calmodulin-mediated pathway. Confocal imaging of living ARPE-19 cells expressing TRMA-associated hTHTR-1 mutants (D93H, S143F and G172D) showed various expression phenotypes. These results demonstrate for the first time the existence of a specialized and regulated uptake process for thiamine in a cellular model of human retinal pigment epithelia that involves hTHTR-1 and hTHTR-2. Further, clinically relevant mutations in hTHTR-1 lead to impaired cell surface expression or function of the transporter in retinal epithelial ARPE-19 cells. PMID- 17463048 TI - The up and down bobbing of human walking: a compromise between muscle work and efficiency. AB - Human walking has a peculiar straight-legged style. Consequently, the body's centre of mass (CM) moves up and down with each step, which is noticeable in their up and down head bobbing while walking. This vertical CM movement enables humans to save energy via a pendulum-like mechanism but is probably a relatively recent locomotor innovation insofar as earliest bipeds may have walked flexed and flat. We investigated the mechanics, energetics, muscle efficiency and optimization of human walking by decreasing and increasing the vertical CM displacement (flat and bouncy walking) in comparison to normal walking at six speeds (1-6 km h(-1)). In both flat and bouncy walking, the pendular mechanism was reduced and the energy cost was increased. However, this increase was unexpectedly much sharper in flat walking where muscles provided normal mechanical work but with a decrease in muscle efficiency. In bouncy walking, muscles provided extra mechanical work in an efficient way. Our results showed that not only do humans bob up and down in normal walking to save energy via a pendulum-like mechanism but also to make their muscles work efficiently. Actually, walking flat makes the muscles work in unfavourable conditions that waste energy. Furthermore, we are still close to a flat CM displacement relative to our current ability to change this displacement, which suggests that reducing vertical CM displacement is indeed important but only to certain limits. Evolution may ultimately have chosen the best compromise between flat locomotion that requires little work to move and bouncy locomotion that improves muscle efficiency to minimize energy consumption. PMID- 17463049 TI - Cell type-specific relationships between spiking and [Ca2+]i in neurons of the Xenopus tadpole olfactory bulb. AB - Multi-neuronal recordings with Ca2+ indicator dyes usually relate [Ca2+]i to action potentials (APs) assuming a stereotypical dependency between the two. However, [Ca2+]i affects and is affected by numerous complex mechanisms that differ from cell type to cell type, from cell compartment to cell compartment. Moreover, [Ca2+]i depends on the specific way a cell is activated. Here we investigate, by combining calcium imaging and on-cell patch clamp recordings, the relationship between APs (spiking) and somatic [Ca2+]i in mitral and granule cells of the olfactory bulb in Xenopus laevis tadpoles. Both cell types exhibit ongoing and odour-modulated [Ca2+]i dynamics. In mitral cells, the occurrence of APs in both spontaneous and odour-evoked situations correlates tightly to step like [Ca2+]i increases. Moreover, odorant-induced suppression of spontaneous firing couples to a decrease in [Ca2+]i. In contrast, granule cells show a substantial number of uncorrelated events such as increases in [Ca2+]i without APs occurring or APs without any effect upon [Ca2+]i. The correlation between spiking and [Ca2+]i is low, possibly due to somatic NMDAR-mediated and subthreshold voltage-activated Ca2+ entries, and thus does not allow a reliable prediction of APs based on calcium imaging. Taken together, our results demonstrate that the relationship between somatic [Ca2+]i and APs can be cell type specific. Taking [Ca2+]i dynamics as an indicator for spiking activity is thus only reliable if the correlation has been established in the system of interest. When [Ca2+]i and APs are precisely correlated, fast calcium imaging is an extremely valuable tool for determining spatiotemporal patterns of APs in neuronal population. PMID- 17463050 TI - Ten largest racial and ethnic health disparities in the United States based on Healthy People 2010 Objectives. AB - A consistent framework has been developed for measuring health disparities and making comparisons across indicators with regard to the public health goals of Healthy People 2010. Disparities are measured as the percent difference from the best group rate, with all indicators being expressed in terms of adverse events. The 10 largest health disparities for each of five US racial and ethnic groups are identified here. There are both similarities and differences in the largest health disparities. New cases of tuberculosis and drug-induced death rates are among the largest health disparities for four of the five racial and ethnic groups. However, drug-induced death is the only indicator among the 10 largest disparities that is shared by both Black and White non-Hispanic populations. PMID- 17463051 TI - A grape berry (Vitis vinifera L.) cation/proton antiporter is associated with berry ripening. AB - We have cloned and characterized VvNHX1, a gene encoding a vacuolar cation/H(+) antiporter from Vitis vinifera cv. Cabernet Sauvignon. VvNHX1 belongs to the vacuolar NHX protein family and showed high similarity to other known vacuolar antiporters. The expression of VvNHX1 partially complements the salt- and hygromycin-sensitive phenotypes of an ena1-4 nhx1 yeast strain. Immunoblots of vacuoles of yeast expressing a VvNHX1, together with the expression of a VvNHX1 GFP (green fluorescent protein) chimera demonstrated that VvNHX1 localized to the vacuoles. VvNHX1 displayed low affinity K(+)/H(+) and Na(+)/H(+) exchange activities (12.8 and 40.2 mM, respectively). The high levels of expression of VvNHX1 during the veraison and post-veraison stages would indicate that the increase in vacuolar K(+) accumulation, mediated by VvNHX1, is needed for vacuolar expansion. This process, together with the rapid accumulation of reducing sugars, would drive water uptake to the berry and the concomitant berry size increase, typical of the post-veraison stage of growth. PMID- 17463052 TI - Functional analysis of the essential bifunctional tobacco enzyme 3-dehydroquinate dehydratase/shikimate dehydrogenase in transgenic tobacco plants. AB - In plants, the shikimate pathway occurs in the plastid and leads to the biosynthesis of aromatic amino acids. The bifunctional 3-dehydroquinate dehydratase/shikimate dehydrogenase (DHD/SHD) catalyses the conversion of dehydroquinate into shikimate. Expression of NtDHD/SHD was suppressed by RNAi in transgenic tobacco plants. Transgenic lines with <40% of wild-type activity displayed severe growth retardation and reduced content of aromatic amino acids and downstream products such as cholorogenic acid and lignin. Dehydroquinate, the substrate of the enzyme, accumulated. However, unexpectedly, so did the product, shikimate. To exclude that this finding is due to developmental differences between wild-type and transgenic plants, the RNAi approach was additionally carried out using a chemically inducible promoter. This approach revealed that the accumulation of shikimate was a direct effect of the reduced activity of NtDHD/SHD with a gradual accumulation of both dehydroquinate and shikimate following induction of gene silencing. As an explanation for these findings the existence of a parallel extra-plastidic shikimate pathway into which dehydroquinate is diverted is proposed. Consistent with this notion was the identification of a second DHD/SHD gene in tobacco (NtDHD/SHD-2) that lacked a plastidic targeting sequence. Expression of an NtDHD/SHD-2-GFP fusion revealed that the NtDHD/SHD-2 protein is exclusively cytosolic and is capable of shikimate biosynthesis. However, given the fact that this cytosolic shikimate synthesis cannot complement loss of the plastidial pathway it appears likely that the role of the cytosolic DHD/SHD in vivo is different from that of the plastidial enzyme. These data are discussed in the context of current models of plant intermediary metabolism. PMID- 17463053 TI - Hepatic gene expression changes in hypothyroid juvenile mice: characterization of a novel negative thyroid-responsive element. AB - The molecular mechanisms involved in the response of developing mice to disruptions in maternal thyroid hormone (TH) homeostasis are poorly characterized. We used DNA microarrays to examine a broad spectrum of genes from the livers of mice rendered hypothyroid by treating pregnant mice from gestational d 13 to postnatal d 15 with 6-propyl-2-thiouracil in drinking water. Twenty-four individuals (one male and one female pup from six litters of control or 6-propyl-2-thiouracil treatment groups, respectively) were profiled using Agilent oligonucleotide microarrays. MAANOVA identified 96 differentially expressed genes (false discovery rate adjusted P < 0.1 and fold change > 2 in at least one gender). Of these, 72 genes encode proteins of known function, 15 of which had previously been identified as regulated by TH. Pathway analysis revealed these genes are involved in metabolism, development, cell proliferation, apoptosis, and signal transduction. An immediate-early response gene, Nr4a1 (nuclear receptor subfamily 4, group A, member 1), was up-regulated by 3-fold in hypothyroid juvenile mouse liver; treatment of HepG2 cells with T(3) resulted in down-regulation of Nr4a1. A potential thyroid response element -1218 to -1188 bp upstream of the promoter region of Nr4a1 was identified and demonstrated to bind TH receptor (TR)-alpha and TRbeta. Point mutation or deletion of the sequence containing the potential Nr4a1-thyroid response element in transient gene expression studies resulted in both higher basal expression and loss of T(3) regulatory capacity, suggesting that this site is responsible for the negative regulation of gene expression by TR and TH. PMID- 17463054 TI - Androgen-dependent regulation of brain-derived neurotrophic factor and tyrosine kinase B in the sexually dimorphic spinal nucleus of the bulbocavernosus. AB - Castration of adult male rats causes the dendrites of androgen-sensitive motoneurons of the spinal nucleus of the bulbocavernosus (SNB) to retract. Brain derived neurotrophic factor (BDNF), via activation of tyrosine receptor kinase B (trkB), has been implicated in mediating androgen effects on SNB dendrites. We used in situ hybridization to demonstrate that SNB motoneurons in gonadally intact adult male rats contain mRNA for both BDNF and trkB. Two weeks after gonadectomy, both transcripts were significantly decreased in SNB motoneurons but not in the non-androgen-responsive motoneurons of the adjacent retrodorsolateral nucleus (RDLN). In a second experiment, target perineal and foot muscles of SNB and RDLN motoneurons, respectively, were injected with the retrograde tracer Fluorogold, and then immunocytochemistry was performed to examine the distribution of BDNF and trkB proteins in SNB and RDLN motoneurons and their glutamatergic afferents. Confocal analysis revealed that gonadectomy induces a loss of BDNF protein in SNB dendrites but not in RDLN dendrites. Testosterone treatment of castrates prevented the loss of BDNF from SNB dendrites. Confocal analysis also revealed trkB protein in SNB and RDLN dendrites and in their glutamatergic afferents. Gonadectomy had no discernable effect on trkB protein in SNB or RDLN motoneurons or in their glutamatergic afferents. These results suggest that androgen maintains a BDNF-signaling pathway in SNB motoneurons that may underlie the maintenance of dendritic structure and synaptic signaling. PMID- 17463055 TI - Female mice haploinsufficient for an inactivated androgen receptor (AR) exhibit age-dependent defects that resemble the AR null phenotype of dysfunctional late follicle development, ovulation, and fertility. AB - The role of classical genomic androgen receptor (AR) mediated actions in female reproductive physiology remains unclear. Female mice homozygous for an in-frame deletion of exon 3 of the Ar (AR(-/-)) were subfertile, exhibiting delayed production of their first litter (AR(+/+) = 22 d vs. AR(-/-) = 61 d, P < 0.05) and producing 60% fewer pups/litter (AR(+/+): 8.1 +/- 0.4 vs. AR(-/-): 3.2 +/- 0.9, P < 0.01). Heterozygous females (AR(+/-)) exhibited an age-dependent 55% reduction (P < 0.01) in pups per litter, evident from 6 months of age (P < 0.05), compared with AR(+/+), indicating a significant gene dosage effect on female fertility. Ovulation was defective with a significant reduction in corpora lutea numbers (48-79%, P < 0.01) in 10- to 12- and 26-wk-old AR(+/-) and AR(-/-) females and a 57% reduction in oocytes recovered from naturally mated AR(-/-) females (AR(+/+): 9.8 +/- 1.0 vs. AR(-/-): 4.2 +/- 1.2, P < 0.01); however, early embryo development to the two-cell stage was unaltered. The delay in first litter, reduction in natural ovulation rate, and aromatase expression in AR(+/-) and AR(-/-) ovaries, coupled with the restored ovulation rate by gonadotropin hyperstimulation in AR(-/-) females, suggest aberrant gonadotropin regulation. A 2.7-fold increase (AR(+/+): 35.4 +/- 13.4 vs. AR(-/-): 93.9 +/- 6.1, P < 0.01) in morphologically unhealthy antral follicles demonstrated deficiencies in late follicular development, although growing follicle populations and growth rates were unaltered. This novel model reveals that classical genomic AR action is critical for normal ovarian function, although not for follicle depletion and that haploinsufficiency for an inactivated AR may contribute to a premature reduction in female fecundity. PMID- 17463056 TI - Ligand-activated peroxisome proliferator activated receptor gamma alters placental morphology and placental fatty acid uptake in mice. AB - The nuclear receptor peroxisome proliferator activated receptor gamma (PPARgamma) is essential for murine placental development. We previously showed that activation of PPARgamma in primary human trophoblasts enhances the uptake of fatty acids and alters the expression of several proteins associated with fatty acid trafficking. In this study we examined the effect of ligand-activated PPARgamma on placental development and transplacental fatty acid transport in wild-type (wt) and PPARgamma(+/-) embryos. We found that exposure of pregnant mice to the PPARgamma agonist rosiglitazone for 8 d (embryonic d 10.5-18.5) reduced the weights of wt, but not PPARgamma(+/-) placentas and embryos. Exposure to rosiglitazone reduced the thickness of the spongiotrophoblast layer and the surface area of labyrinthine vasculature, and altered expression of proteins implicated in placental development. The expression of fatty acid transport protein 1 (FATP1), FATP4, adipose differentiation related protein, S3-12, and myocardial lipid droplet protein was enhanced in placentas of rosiglitazone treated wt embryos, whereas the expression of FATP-2, -3, and -6 was decreased. Additionally, rosiglitazone treatment was associated with enhanced accumulation of the fatty acid analog 15-(p-iodophenyl)-3-(R, S)-methyl pentadecanoic acid in the placenta, but not in the embryos. These results demonstrate that in vivo activation of PPARgamma modulates placental morphology and fatty acid accumulation. PMID- 17463057 TI - VGF-derived peptide, TLQP-21, regulates food intake and body weight in Siberian hamsters. AB - The Siberian hamster survives winter by decreasing food intake and catabolizing abdominal fat reserves, resulting in a sustained, profound loss of body weight. VGF gene expression is photoperiodically regulated in the hypothalamus with significantly higher expression in lean Siberian hamsters. The aim of this study was to investigate the role of VGF in regulating these seasonal cycles by determining the effects of a VGF-derived peptide (TLQP-21) on food intake and body weight. Acute intracerebroventricular administration of TLQP-21 decreased food intake, and chronic treatment caused a sustained reduction in food intake and body weight and decreased abdominal fat depots. Behavioral analysis revealed that TLQP-21 reduced meal size but not the frequency of feeding bouts, suggesting a primary action on satiety. Hamsters treated with TLQP-21 lost a similar amount of weight as a pair-fed group in which food intake was matched to that of the TLQP-21-treated group. Central or peripheral treatment with TLQP-21 did not produce a significant effect on resting metabolic rate. We conclude that the primary action of TLQP-21 is to decrease food intake rather than increase energy expenditure. TLQP-21 treatment caused a decrease in UCP-1 mRNA in brown adipose tissue, but hypothalamic expression of orexigenic and anorexigenic neuropeptide genes remained unchanged after TLQP-21 treatment, although compensatory increases in NPY and AgRP mRNA were observed in the pair-fed hamsters. The effects of TLQP 21 administration are similar to those in hamsters in short days, suggesting that increased VGF activity may contribute to the hypophagia that underlies the seasonal catabolic state. PMID- 17463058 TI - Involvement of neuropeptide Y Y1 receptors in the regulation of neuroendocrine corticotropin-releasing hormone neuronal activity. AB - The neuroendocrine parvocellular CRH neurons in the paraventricular nucleus (PVN) of the hypothalamus are the main integrators of neural inputs that initiate hypothalamic-pituitary-adrenal (HPA) axis activation. Neuropeptide Y (NPY) expression is prominent within the PVN, and previous reports indicated that NPY stimulates CRH mRNA levels. The purpose of these studies was to examine the participation of NPY receptors in HPA axis activation and determine whether neuroendocrine CRH neurons express NPY receptor immunoreactivity. Infusion of 0.5 nmol NPY into the third ventricle increased plasma corticosterone levels in conscious rats, with the peak of hormone levels occurring 30 min after injection. This increase was prevented by pretreatment with the Y1 receptor antagonist BIBP3226. Immunohistochemistry showed that CRH-immunoreactive neurons coexpressed Y1 receptor immunoreactivity (Y1r-ir) in the PVN, and a majority of these neurons (88.8%) were neuroendocrine as determined by ip injections of FluoroGold. Bilateral infusion of the Y1/Y5 agonist, [leu(31)pro(34)]NPY (110 pmol), into the PVN increased c-Fos and phosphorylated cAMP response element-binding protein expression and elevated plasma corticosterone levels. Increased expression of c Fos and phosphorylated cAMP response element-binding protein was observed in populations of CRH/Y1r-ir cells. The current findings present a comprehensive study of NPY Y1 receptor distribution and activation with respect to CRH neurons in the PVN. The expression of NPY Y1r-ir by neuroendocrine CRH cells suggests that alterations in NPY release and subsequent activation of NPY Y1 receptors plays an important role in the regulation of the HPA. PMID- 17463059 TI - Lipin1 regulation by estrogen in uterus and liver: implications for diabetes and fertility. AB - Estrogens are essential for fertility and also have important effects on regulation of adiposity and the euglycemic state. We report here that lipin1, a candidate gene for lipodystrophy and obesity that is a phosphatidic acid phosphatase critical in regulation of cellular levels of diacylglycerol and triacylglycerol and a key regulator of lipid utilization, is rapidly and robustly down-regulated in the uterus by estradiol via the estrogen receptor. Lipin1 is expressed predominantly in the uterine luminal and glandular epithelium, and during the estrous cycle, lipin1 is lowest when blood levels of estrogen are highest. Lipin1 is expressed throughout all cells in the liver of ovariectomized female mice, and a sustained down-regulation is observed at the mRNA, protein and immunohistochemical levels after estrogen administration. Because the coupling of proper energy use and availability is central to reproduction, we also investigated expression of lipin1 in the uterus and liver of several mouse models of diabetes. Nonobese diabetic (NOD) mice, which have high blood levels of estrogen and impaired fertility, were severely deficient in lipin1 in the uterus and liver, which, interestingly, could be restored by insulin treatment. By contrast, nonobese diabetic/severe combined immunodeficient (NOD-SCID) mice, which do not develop diabetes, showed normal levels of lipin1. Our findings of lipin1 regulation by estrogen in two key target organs suggest a new role for this lipid-regulating phosphatase not only in central metabolic regulation but also in uterine function and reproductive biology. Estrogen regulation of lipin1 may provide a mechanistic link between estrogens, lipid metabolism, and lipid signaling. PMID- 17463060 TI - Prolactin induces regional vasoconstriction through the beta2-adrenergic and nitric oxide mechanisms. AB - Prolactin has been associated with many effects and has been implicated in the pathogenesis of pregnancy-related hypertensive disorders, although little is known about its vascular effects. The present study was designed to determine the primary effect of prolactin on regional vascular beds and the mechanisms involved. In 37 anesthetized pigs, the infusion of 0.17 mug/kg min of prolactin at constant heart rate and arterial pressure decreased coronary, mesenteric, renal, and iliac blood flow. This response was graded in further five pigs by increasing the infused dose of the hormone between 0.017 and 1 mug/kg min. In 22 of the 37 pigs, blockade of cholinergic receptors (five pigs) and of alpha adrenoceptors (five pigs) did not affect the prolactin-induced vascular response, which was abolished by blockade of beta(2)-adrenoceptors (five pigs) and by blockade of vascular nitric oxide (NO) synthase (seven pigs). In 15 of the 37 pigs the increases in measured blood flows caused by iv infusion of isoproterenol (five pigs) and by intraarterial administration of acetylcholine (five pigs) and of sodium nitroprusside (five pigs) were significantly reduced by infusion of prolactin. Moreover, the treatment of porcine aortic endothelial cells by prolactin caused a reduction of NO production and of the phosphorylation of ERK, Akt, and p38, which was prevented by the concomitant treatment by the beta(2) adrenergic agonist albuterol. The present study showed that iv infusion of prolactin primarily caused coronary, mesenteric, renal, and iliac vasoconstriction. These effects were brought about by the inhibition of a vasodilatory beta(2)-adrenergic receptor-mediated effect related to the NO intracellular pathway. PMID- 17463061 TI - Permethrin may disrupt testosterone biosynthesis via mitochondrial membrane damage of Leydig cells in adult male mouse. AB - Permethrin, a popular synthetic pyrethroid insecticide used to control noxious insects in agriculture, forestry, households, horticulture, and public health throughout the world, poses risks of environmental exposure. Here we evaluate the reproductive toxicity of cis-permethrin in adult male ICR mice that were orally administered cis-permethrin (0, 35, or 70 mg/kg d) for 6 wk. Caudal epididymal sperm count and sperm motility in the treated groups were statistically reduced in a dose-dependent manner. Testicular testosterone production and plasma testosterone concentration were significantly and dose-dependently decreased with an increase in LH, and a significant regression was observed between testosterone levels and cis-permethrin residues in individual mice testes after exposure. However, no significant changes were observed in body weight, reproductive organ absolute and relative weights, sperm morphology, and plasma FSH concentration after cis-permethrin treatment. Moreover, cis-permethrin exposure significantly diminished the testicular mitochondrial mRNA expression levels of peripheral benzodiazepine receptor (PBR), steroidogenic acute regulatory protein (StAR), and cytochrome P450 side-chain cleavage (P450scc) and enzyme and protein expression levels of StAR and P450scc. At the electron microscopic level, mitochondrial membrane damage was found in Leydig cells of the exposed mouse testis. Our results suggest that the insecticide permethrin may cause mitochondrial membrane impairment in Leydig cells and disrupt testosterone biosynthesis by diminishing the delivery of cholesterol into the mitochondria and decreasing the conversion of cholesterol to pregnenolone in the cells, thus reducing subsequent testosterone production. PMID- 17463062 TI - Corticotropin-releasing hormone receptor type 1 and type 2 mediate differential effects on 15-hydroxy prostaglandin dehydrogenase expression in cultured human chorion trophoblasts. AB - Throughout gestation, the chorion laeve controls the levels of biologically active prostaglandins (PGs) by its high level of nicotinamide adenine dinucleotide-dependent 15-hydroxy PG dehydrogenase (PGDH). In this study, we investigate the effects mediated by CRH receptors on the expression of PGDH in the chorion. We found that both CRHR1 and CRHR2 were localized in cultured chorion trophoblast cells, with CRH-R1alpha, R1beta, R1c, R1e, and R1f and CRHR2beta isoforms identified in these cells. To block the actions of endogenous CRH and its related peptides, cultured chorion trophoblasts were treated with an increasing concentration of alpha-helical CRH 9-41, the nonselective CRH receptor antagonist, which resulted in decreased mRNA and protein expression as well as the activity of PGDH. To investigate the individual role of CRHR1 and CRHR2, cell cultures were treated with the specific CRHR1 antagonist antalarmin and CRHR2 antagonist astressin2B, respectively. The results showed that antalarmin increased whereas astressin2B decreased mRNA and protein expression as well as the activity of PGDH in chorion cells. When the cells were treated with an exclusive CRHR2 agonist, urocortin II, elevated expression and activity of PGDH was exhibited. However, cells treated with either exogenous CRH or urocortin I showed significantly increased PGDH expression, and these effects could be blocked by astressin2B but not by antalarmin. We suggest that, in chorion trophoblast cells, CRHR1 and CRHR2 mediate divergent effects on PGDH expression, and this may provide a precise regulation of PGs levels from chorion to myometrium during pregnancy. PMID- 17463063 TI - Administration of adrenocorticotropic hormone during chicken embryonic development prematurely induces pituitary growth hormone cells. AB - Treatment of fetal rats and embryonic chickens with exogenous glucocorticoids induces premature GH cell differentiation. However, it is unknown whether the developing adrenal gland is capable of mounting this response autonomously. The present study determined whether stimulation of the adrenal gland in developing chicken embryos through administration of ACTH could induce a premature increase in GH cells. We found that plasma corticosterone and ACTH levels increased between embryonic day (e) 11 and e17, consistent with GH cell (somatotroph) ontogeny. Injection of ACTH into eggs on e9, e10, or e11 increased somatotrophs on e14. In contrast, thyroid-stimulating hormone, CRH, alpha-MSH, GHRH, and TRH were ineffective. Culture of e11 pituitary cells with ACTH failed to induce somatotrophs, suggesting an indirect action of ACTH on GH cells in vivo. Intravenous administration of ACTH dramatically increased plasma levels of corticosterone within 1 h and increased the percentage of pituitary somatotrophs within 24 h. Although ACTH administration increased the relative abundance of pituitary GH cells, there was no effect on plasma levels of GH, IGF-I, or IGF-II, or in hepatic expression of IGF-I or IGF-II mRNA. We conclude that ACTH administration can increase the population of GH cells in the embryonic pituitary. However, this treatment alone does not lead to downstream activation of hepatic IGF production. These findings indicate that the embryonic adrenal gland, and ultimately anterior pituitary corticotrophs, may function to regulate pituitary GH cell differentiation during embryonic development. PMID- 17463064 TI - A NavX-guided ablation of a nodo-fascicular fibre. AB - The NavX system was used to guide mapping and successful ablation of a reciprocating antidromic tachycardia involving anterograde conduction over a nodo fascicular fibre and retrograde conduction over the His bundle and AV node. This novel mapping system allowed visualization of the lower insertion site of the nodo-fascicular accessory pathway approximately 1.5-2 cm away from the tricuspid annulus in a three-dimensional reconstruction of the right atrium and right ventricle. PMID- 17463065 TI - The Arg389Gly beta1-adrenergic receptor gene polymorphism and susceptibility to faint during head-up tilt test. AB - AIMS: To investigate the association of the Gly389 allele with positive head-up tilt test (HUT) in a Mexican Mestizo population. METHODS AND RESULTS: HUT results were compared between carriers (one or two copies of the Gly389 allele) and non carriers (Arg389Arg genotype) of the Gly389 allele of the beta(1)AR gene in 50 patients with unexplained syncope. Thirty-three patients (66%) had a positive HUT. Patients with a positive HUT had a higher Gly389 allele frequency compared with those with a negative test (30.3 vs. 3%; OR 13; pC = 0.012). Moreover, when comparing positive HUT in passive drug-free phase, positive HUT in pharmacological (nitrate) phase, and negative (both phases), a decreasing gradient in the frequencies of the Gly389 allele was found among the three groups: 45.4, 22.7, and 3%, respectively. CONCLUSION: An association of positive tilt table testing to a single nucleotide polymorphism with a Gly to Arg switch at position 389 of the beta(1)AR was found. This polymorphism may contribute to susceptibility to faint during orthostatic challenge. PMID- 17463066 TI - Multiple sclerosis immunopathic trait and HLA-DR(2)15 as independent risk factors in multiple sclerosis. AB - We analysed HLA haplotypes in pairs of 78 sporadic multiple sclerosis (MS) patients and 78 healthy siblings. The presence of 2 oligoclonal IgG bands, detected by immunoblotting of the cerebrospinal fluid in healthy siblings, has previously been defined as MS immunopathic trait (MSIT), based on a cut-off derived from healthy unrelated volunteers. The frequency of MSIT was 17.9% (n=14/78 siblings). The HLA-DR(15)2 allelle was present in 21.4% (n=3/14) of the siblings with MSIT, in 40.6% (n =26/64) of the siblings without MSIT, and in 59% (n =46/78) of the patients with clinically-definite (CD) MS. The distribution of zero, one or two HLA-DR(2)15 alleles was significantly skewed towards a lower allelle count in the siblings with MSIT compared with the group of unrelated siblings with MS (P=0.002), and also lower than their related siblings with MS (P=0.1). These results suggest that the MS susceptibility gene, HLA-DR(2)15 type, does not induce MSIT, and conceivably these are two separate risk factors in the development of MS. The effect of HLA-DR(2)15 and MSIT in sporadic MS appears to be synergistic. PMID- 17463067 TI - Glyoxalase I A111E, paraoxonase 1 Q192R and L55M polymorphisms: susceptibility factors of multiple sclerosis? AB - Multiple sclerosis (MS) is characterized by chronic inflammation and demyelination of the central nervous system (CNS). Accumulating data indicate that oxidative stress, leading to reactive oxygen species (ROS) production and lipid peroxidation, as well as elevated levels of advanced glycation end products (AGE) in CNS neurons, might play a pivotal role in the pathogenesis of a number of diseases with a neurodegenerative aspect, such as MS. Therefore, polymorphisms of genes encoding endogenous free-radical scavenging systems, such as paraoxonase 1 (PON1), and anti-glycation defences, such as glyoxalase I (GI), could influence susceptibility to MS. In the present study, we have undertaken a case-control study to investigate the possible association of GI A111E, PON1 Q192R and L55M polymorphisms with the risk of MS. The three polymorphisms were characterized in 209 patients with relapsing-remitting MS (RRMS) and in 213 healthy controls by PCR/RFLP methods using DNA from lymphocytes. We found that individuals with the GI/AE-EE genotypes and PON55/LM-MM genotypes had a significantly higher risk of MS compared with the other genotypes. The two polymorphisms appear to be common genetic traits that are associated with an increased risk for MS--the analysis of both, in each single case, may be a revealing predictable factor for MS risk. PMID- 17463068 TI - UCP2 and mitochondrial haplogroups as a multiple sclerosis risk factor. AB - In the actual scenario of the search for further genetic susceptibility factors, a recent paper noted an SNP in the UCP2 gene as a multiple sclerosis (MS) risk factor. UCP2 is a member of the mitochondrial proton transport family, which uncouples proton entry in the mitochondrial matrix from ATP synthesis. mtDNA haplogroups are also associated with ATP production, and are linked with mitochondrial proton transport. In this work, we studied the UCP2 SNP and the mitochondrial haplogroups distribution in a Spanish MS population, with a population sub-group of Basque-origin patients. Our results confirm the link between UCP2 SNP and MS, and show a slight relation between this SNP and mitochondrial haplogroups. PMID- 17463069 TI - Interferon-beta regulates cytokines and BDNF: greater effect in relapsing than in progressive multiple sclerosis. AB - The mechanism of action of interferon (IFN)-beta therapy in multiple sclerosis (MS) is only partially known, and its efficacy changes with disease stage. In different forms of MS, we determined how IFN-beta regulates mononuclear cell production of the important anti-inflammatory Th2 cytokine - IL-10, the Th1 cytokine - IFN-gamma, and the brain-derived neurotrophic protein - BDNF. Activated T cells and monocytes from therapy-naive patients secreted more IL-10 than healthy controls. During IFN-beta therapy, however, T cells produced less IL 10. In vitro, IFN-beta stimulated IL-10 production by activated T cells, but inhibited IL-10 secretion by activated monocytes, a richer source of IL-10 than T cells. The form of MS also affected cytokine production. IL-10 and BDNF levels in MNC were high during relapsing/remitting (RR) MS, but low in progressive MS. Surprisingly, IFN-beta therapy increased BDNF levels in antidepressant-naive patients, but BDNF was lower during concurrent antidepressant drug therapy, suggesting an interaction between MS, depression, and neurodegeneration. IFN-beta in vitro strongly induced IL-10 and IFN-gamma in activated T cells in RRMS, but not in progressive MS, suggesting IFN resistance. IFN-beta effects are specific for disease state and immune subsets, possibly explaining why IFN-beta therapy is most effective in early T cell-regulated RRMS, but less beneficial in progressive MS, where chronic plaques contain few T cells and high numbers of monocytes. PMID- 17463070 TI - Histopathology and serial, multimodal magnetic resonance imaging in a multiple sclerosis variant. AB - Defining tools in magnetic resonance imaging (MRI) representing specific pathological processes is needed to understand the complex relationship between inflammation, myelin breakdown, axonal injury and clinical symptoms in multiple sclerosis (MS) and its variants. Here, we describe a case of histologically defined MS, in which the radiological appearance of the lesion and clinical course support the diagnosis of Balo's concentric sclerosis. Serial magnetization transfer, diffusion tensor imaging and 1H-magnetic resonance spectroscopy, from 14 days to 13 months after biopsy, allow the contextual interpretation of specific pathological changes. In our case, acute inflammation was sensitively traced by fractional anisotropy and increased lactate in spectroscopy. In contrast, magnetization transfer ratio and the apparent diffusion coefficient monitor the sequential loss of tissue in selected rings of the lesion. The delay from the peak of symptoms in a dramatic clinical course to the maximum tissue destruction indicated through MRI suggests that compromise of axonal function may be decisive for the acute clinical situation. This is the first report comparing 1H-magnetic resonance spectroscopy, magnetization transfer and diffusion tensor imaging with histopathology in a patient with Balo's concentric sclerosis. PMID- 17463071 TI - Voxel-based analysis of grey matter magnetization transfer ratio maps in early relapsing remitting multiple sclerosis. AB - Previous studies using magnetization transfer ratio (MTR) histogram analysis have demonstrated the existence of global grey matter (GM) abnormalities in patients with early relapsing-remitting multiple sclerosis (RRMS). However, MTR histogram analysis does not provide any information on the localization of the morphological changes within the GM. The aim of this study was to investigate the localization of GM injury in early RRMS, performing voxel-based analysis of GM MTR maps. Statistical mapping analysis of GM MTR maps was performed in a group of 38 patients with early RRMS and 45 healthy controls. Between-group comparisons (P<0.05, corrected for multiple comparisons) demonstrated significant GM MTR decrease in patients located in the bilateral lenticular nuclei, the bilateral insula, the left posterior cingulate cortex, and the right orbitofrontal cortex. To limit the potential confounding effect of regional GM atrophy, the percentages of GM were assessed in the regions showing significant MTR decrease, and no GM atrophy was evidenced in these regions. This study demonstrates that several GM regions are commonly affected in patients with early RRMS. Predominant involvement of these structures may be partly related to their vulnerability to anterograde or retrograde degeneration from transected axons in the white matter and/or to the predominant localization of GM demyelinating lesions in such regions. PMID- 17463072 TI - Interferon beta-1a slows progression of brain atrophy in relapsing-remitting multiple sclerosis predominantly by reducing gray matter atrophy. AB - BACKGROUND: Brain atrophy, as assessed by magnetic resonance imaging (MRI), has been correlated with disability in patients with multiple sclerosis (MS). Recent evidence indicates that both white matter (WM) and gray matter (GM) are subject to atrophy in patients with MS. Although neurological deficiencies in MS are primarily due to loss of WM, the clinical significance of GM atrophy has not been fully explored in MS. METHODS: We have undertaken a three-year, open-label study, comparing 26 patients who elected to receive intramuscular interferon beta-1a (IFN beta-1a) therapy, with 28 patients who elected not to receive therapy. Both groups had quantitative cranial MRI scans at study entry and after three years, and standardized clinical assessments every six months. Brain parenchymal fraction (BPF), GM fraction (GMF), and WM fraction (WMF) percent changes were calculated, and T2- and T1-lesion volumes (LVs) assessed. RESULTS: After three years, mean percent (%) change in BPF favored the IFN beta-1a treatment group (IFN beta-1a -1.3% versus the control group -2.5%, P=0.009), as did the mean percent change in GMF (+0.2 versus -1.4%, P=0.014), and the mean percent change in T1-LV (-9.3 versus +91.6%, P=0.011). At the end of the study, there was a significant within-patient decrease in BPF for both groups (P=0.02 for the IFN beta-1a treatment group, and P<0.001 for the control group), a significant within patient decrease in WMF for the IFN beta-1a treatment group (P=0.01), and a significant decrease in GMF for the control group (P=0.013) when compared with baseline. CONCLUSION: Over a three-year period, treatment with IFN beta-1a significantly slowed the progression of whole-brain and GM atrophy, and of T1 hypointense LV accumulation, when compared with the control group. PMID- 17463073 TI - Pooled historical MRI data as a basis for research in multiple sclerosis--a statistical evaluation. AB - Pooled data from placebo groups of different trials can serve as historical control for ongoing or future therapeutic studies and as a reference for power calculations. In order to assess their usefulness for this purpose, we investigated the degree of heterogeneity of placebo arm data from 14 controlled studies included in the database of the Sylvia Lawry Centre for Multiple Sclerosis Research. Since different criteria for the inclusion/exclusion of patients were used in these studies, an attempt was made to adjust the distribution of magnetic resonance imaging (MRI) measures for the differences in the study populations. The analyses showed that, even after adjustment, significant differences remained. This heterogeneity does not reduce the usefulness of the database for statistical analysis of inter-relationships between variables, provided that it is explicitly taken into account as a stratification factor. However, care must be taken when trying to compare the results of a newly treated group with the patients of this pool. Heterogeneity in some MRI variables was greatly reduced when only studies from the same image analysis centre were compared. PMID- 17463074 TI - The clinical response to minocycline in multiple sclerosis is accompanied by beneficial immune changes: a pilot study. AB - Minocycline has immunomodulatory and neuroprotective activities in vitro and in an animal model of multiple sclerosis (MS). We have previously reported that minocycline decreased gadolinium-enhancing activity over six months in a small trial of patients with active relapsing-remitting MS (RRMS). Here we report the impact of oral minocycline on clinical and magnetic resonance imaging (MRI) outcomes and serum immune molecules in this cohort over 24 months of open-label minocycline treatment. Despite a moderately high pretreatment annualized relapse rate (1.3/year pre-enrolment; 1.2/year during a three-month baseline period) prior to treatment, no relapses occurred between months 6 and 24. Also, despite very active MRI activity pretreatment (19/40 scans had gadolinium-enhancing activity during a three-month run-in), the only patient with gadolinium-enhancing lesions on MRI at 12 and 24 months was on half-dose minocycline. Levels of the p40 subunit of interleukin (IL)-12, which at high levels might antagonize the proinflammatory IL-12 receptor, were elevated over 18 months of treatment, as were levels of soluble vascular cell adhesion molecule-1. The activity of matrix metalloproteinase-9 was decreased by treatment. Thus, clinical and MRI outcomes are supported by systemic immunological changes and call for further investigation of minocycline in MS. PMID- 17463075 TI - The effects of transcutaneous electrical nerve stimulation (TENS) on spasticity in multiple sclerosis. AB - Spasticity is a common and often disabling symptom associated with multiple sclerosis (MS). Transcutaneous electrical nerve stimulation (TENS) has been found effective in reducing spasticity in conditions such as stroke, but there is little evidence to support its use in MS. The aim of this study was to evaluate the effectiveness of TENS on spasticity in MS and, furthermore, to compare two different application times. Thirty-two subjects were randomized into two groups, and a single, blind, crossover design was used to compare two weeks of 60 minutes and 8 hours daily of TENS applications (100 Hz and 0.125 ms pulse width). Outcomes were examined using the Global Spasticity Score (GSS), the Penn Spasm Score (PSS), and a visual analogue scale (VAS) for pain. The results of the study demonstrated that there were no statistically significant differences in the GSS following either 60 minutes or 8 hours daily of TENS (P=0.433 and 0.217, respectively). The 8-hour application time led to a significant reduction in muscle spasm (P=0.038) and pain (P = 0.008). Thus, this study suggests that, whilst TENS does not appear to be effective in reducing spasticity, longer applications may be useful in treating MS patients with pain and muscle spasm. PMID- 17463076 TI - Health, disability, and life insurance experiences of working-age persons with multiple sclerosis. AB - Working-age Americans with multiple sclerosis (MS) may face considerable financial insecurities when they become unable to work and lack the health, disability, and life insurance typically offered through employers. In order to estimate the rates of having these insurance policies, as well as how insurance status affects reports of financial stress, we conducted half-hour telephone interviews with 983 working-age persons across the US, who reported being diagnosed with MS. The interviews occurred from May through November 2005, and among the sampled individuals contacted and confirmed eligible, 93.2% completed the interview. The study population was largely female (78.9%), Caucasian (86.4%), married (68.6%), with at least some college education (71.5%), and unemployed (60.2%). Overall, 96.3% had some health insurance (40.3% with public health insurance, primarily Medicare), 56.7% had long-term disability insurance (36.4% with public programs), and 68.3% had life insurance. Notably, 27.4% indicated that, since being diagnosed with MS, health insurance concerns had significantly affected employment decisions. In addition, 16.4% reported considerable difficulty paying for health care, 27.4% put off or postponed seeking needed health care because of costs, and 22.3% delayed filling prescriptions, skipped medication doses, or split pills because of costs. Overall, 26.6% reported considerable worries about affording even basic necessities, such as food, utilities, and housing. PMID- 17463077 TI - Access to health care for people with multiple sclerosis. AB - The Sonya Slifka Longitudinal Multiple Sclerosis (MS) Study follows a population based cohort of approximately 2000 people with MS to study demographic and clinical characteristics, use and cost of health services, provider and treatment characteristics, neurological, economic, and psychosocial outcomes. We examined key indicators of access to health care and found that the majority of participants had health insurance, a usual source of care, and access to specialty care. Nevertheless, 3.8% did not have health insurance which, with application of sampling weights, corresponds to approximately 7000 people with MS in the US population. Even with insurance, population-based estimates indicated that substantial numbers of people with MS have plans that pay nothing toward prescription medication, limit their access to specialists, and restrict their choice of hospitals and providers. Some 9% of the sample, corresponding to 15,800 people with MS, did not have a usual source of MS care; 11.8% or 17,300 people did not have a usual source of general health care; and 31% or 57,400 people did not see the specialists that they or their physicians wanted them to see. Further, 10.5% or 19,400 people reported difficulty obtaining prescription medication, 4.1% or 7600 people encountered obstacles accessing medical care, and 2.4% or 4500 people could not obtain the mental health services they needed. Finally, out-of-pocket health care expenditures were twice those found for the general population. Two-thirds of study participants (representing almost 70,000 people) chose their MS care providers because they were neurologists or MS specialists, creating a demand that almost certainly exceeds current supply. PMID- 17463078 TI - Anaphylactic reaction to methylprednisolone in multiple sclerosis: a practical approach to alternative corticosteroids. PMID- 17463079 TI - Matrix metalloproteinases (MMP): determination of different forms by different techniques may require different preanalytical strategies. PMID- 17463080 TI - Mapping allosteric connections from the receptor to the nucleotide-binding pocket of heterotrimeric G proteins. AB - Heterotrimeric G proteins function as molecular relays that mediate signal transduction from heptahelical receptors in the cell membrane to intracellular effector proteins. Crystallographic studies have demonstrated that guanine nucleotide exchange on the Galpha subunit causes specific conformational changes in three key "switch" regions of the protein, which regulate binding to Gbetagamma subunits, receptors, and effector proteins. In the present study, nitroxide side chains were introduced at sites within the switch I region of Galphai to explore the structure and dynamics of this region throughout the G protein cycle. EPR spectra obtained for each of the Galpha(GDP), Galpha(GDP)betagamma heterotrimer and Galpha(GTPgammaS) conformations are consistent with the local environment observed in the corresponding crystal structures. Binding of the heterotrimer to activated rhodopsin to form the nucleotide-free (empty) complex, for which there is no crystal structure, causes prominent changes relative to the heterotrimer in the structure of switch I and contiguous sequences. The data identify a putative pathway of allosteric changes triggered by receptor binding and, together with previously published data, suggest elements of a mechanism for receptor-catalyzed nucleotide exchange. PMID- 17463081 TI - Metabolic engineering of Escherichia coli for the production of L-valine based on transcriptome analysis and in silico gene knockout simulation. AB - The L-valine production strain of Escherichia coli was constructed by rational metabolic engineering and stepwise improvement based on transcriptome analysis and gene knockout simulation of the in silico genome-scale metabolic network. Feedback inhibition of acetohydroxy acid synthase isoenzyme III by L-valine was removed by site-directed mutagenesis, and the native promoter containing the transcriptional attenuator leader regions of the ilvGMEDA and ilvBN operon was replaced with the tac promoter. The ilvA, leuA, and panB genes were deleted to make more precursors available for L-valine biosynthesis. This engineered Val strain harboring a plasmid overexpressing the ilvBN genes produced 1.31 g/liter L valine. Comparative transcriptome profiling was performed during batch fermentation of the engineered and control strains. Among the down-regulated genes, the lrp and ygaZH genes, which encode a global regulator Lrp and L-valine exporter, respectively, were overexpressed. Amplification of the lrp, ygaZH, and lrp-ygaZH genes led to the enhanced production of L-valine by 21.6%, 47.1%, and 113%, respectively. Further improvement was achieved by using in silico gene knockout simulation, which identified the aceF, mdh, and pfkA genes as knockout targets. The VAMF strain (Val DeltaaceF Deltamdh DeltapfkA) overexpressing the ilvBN, ilvCED, ygaZH, and lrp genes was able to produce 7.55 g/liter L-valine from 20 g/liter glucose in batch culture, resulting in a high yield of 0.378 g of L-valine per gram of glucose. These results suggest that an industrially competitive strain can be efficiently developed by metabolic engineering based on combined rational modification, transcriptome profiling, and systems-level in silico analysis. PMID- 17463082 TI - Cyclophilin D inactivation protects axons in experimental autoimmune encephalomyelitis, an animal model of multiple sclerosis. AB - Multiple sclerosis (MS) is the leading cause of neurological disability in young adults, affecting some two million people worldwide. Traditionally, MS has been considered a chronic, inflammatory disorder of the central white matter in which ensuing demyelination results in physical disability [Frohman EM, Racke MK, Raine CS (2006) N Engl J Med 354:942-955]. More recently, MS has become increasingly viewed as a neurodegenerative disorder in which neuronal loss, axonal injury, and atrophy of the CNS lead to permanent neurological and clinical disability. Although axonal pathology and loss in MS has been recognized for >100 years, very little is known about the underlying molecular mechanisms. Progressive axonal loss in MS may stem from a cascade of ionic imbalances initiated by inflammation, leading to mitochondrial dysfunction and energetic deficits that result in mitochondrial and cellular Ca2+ overload. In a murine disease model, experimental autoimmune encephalomyelitis (EAE) mice lacking cyclophilin D (CyPD), a key regulator of the mitochondrial permeability transition pore (PTP), developed EAE, but unlike WT mice, they partially recovered. Examination of the spinal cords of CyPD-knockout mice revealed a striking preservation of axons, despite a similar extent of inflammation. Furthermore, neurons prepared from CyPD-knockout animals were resistant to reactive oxygen and nitrogen species thought to mediate axonal damage in EAE and MS, and brain mitochondria lacking CyPD sequestered substantially higher levels of Ca2+. Our results directly implicate pathological activation of the mitochondrial PTP in the axonal damage occurring during MS and identify CyPD, as well as the PTP, as a potential target for MS neuroprotective therapies. PMID- 17463083 TI - Targeted delivery of proteins across the blood-brain barrier. AB - Treatment of many neuronal degenerative disorders will require delivery of a therapeutic protein to neurons or glial cells across the whole CNS. The presence of the blood-brain barrier hampers the delivery of these proteins from the blood, thus necessitating a new method for delivery. Receptors on the blood-brain barrier bind ligands to facilitate their transport to the CNS; therefore, we hypothesized that by targeting these receptors, we may be able to deliver proteins to the CNS for therapy. Here, we report the use of the lentivirus vector system to deliver the lysosomal enzyme glucocerebrosidase and a secreted form of GFP to the neurons and astrocytes in the CNS. We fused the low-density lipoprotein receptor-binding domain of the apolipoprotein B to the targeted protein. This approach proved to be feasible for delivery of the protein and could possibly be used as a general method for delivery of therapeutic proteins to the CNS. PMID- 17463084 TI - NKG2D recognition mediates Toll-like receptor 3 signaling-induced breakdown of epithelial homeostasis in the small intestines of mice. AB - Toll-like receptors (TLRs) and NK receptors are the two most important receptor families in innate immunity. Although it has been observed that TLR signaling can induce or up-regulate the expression of the ligands for stimulatory NK receptors on monocytes or muscle cells, there is not yet a report indicating whether TLR signaling can break down self-tolerance through NK receptors. The present work reports that TLR3 signaling by polyinosinic-polycytidylic acid stimulation induces intestinal epithelial cells (IECs) to express retinoic acid early inducible-1 (a ligand for NKG2D) and to induce NKG2D expression on CD8alphaalpha intestinal intraepithelial lymphocytes by IL-15 derived from TLR3-activated IECs. The blockade of interaction between NKG2D and Rae1 inhibits the cytotoxicity of intraepithelial lymphocytes against IECs in a cell-cell contact-dependent manner and therefore alleviates polyinosinic-polycytidylic acid-induced epithelial destruction and acute mucosal injury of small intestine. These results demonstrate that TLR signaling induces tissue injury through the NKG2D pathway, suggesting that TLR signaling may break down self-tolerance through induction of abnormal expression of ligands for stimulatory NK receptors. PMID- 17463085 TI - Enzymatic analysis of a rhomboid intramembrane protease implicates transmembrane helix 5 as the lateral substrate gate. AB - Intramembrane proteolysis is a core regulatory mechanism of cells that raises a biochemical paradox of how hydrolysis of peptide bonds is accomplished within the normally hydrophobic environment of the membrane. Recent high-resolution crystal structures have revealed that rhomboid proteases contain a catalytic serine recessed into the plane of the membrane, within a hydrophilic cavity that opens to the extracellular face, but protected laterally from membrane lipids by a ring of transmembrane segments. This architecture poses questions about how substrates enter the internal active site laterally from membrane lipid. Because structures are static glimpses of a dynamic enzyme, we have taken a structure-function approach analyzing >40 engineered variants to identify the gating mechanism used by rhomboid proteases. Importantly, our analyses were conducted with a substrate that we show is cleaved at two intramembrane sites within the previously defined Spitz substrate motif. Engineered mutants in the L1 loop and active-site region of the GlpG rhomboid protease suggest an important structural, rather than dynamic, gating function for the L1 loop that was first proposed to be the substrate gate. Conversely, three classes of mutations that promote transmembrane helix 5 displacement away from the protease core dramatically enhanced enzyme activity 4- to 10-fold. Our functional analyses have identified transmembrane helix 5 movement to gate lateral substrate entry as a rate-limiting step in intramembrane proteolysis. Moreover, our mutagenesis also underscores the importance of other residue interactions within the enzyme that warrant further scrutiny. PMID- 17463086 TI - Inhibition of lysine-specific demethylase 1 by polyamine analogues results in reexpression of aberrantly silenced genes. AB - Epigenetic chromatin modification is a major regulator of eukaryotic gene expression, and aberrant epigenetic silencing of gene expression contributes to tumorigenesis. Histone modifications include acetylation, phosphorylation, and methylation, resulting in a combination of histone marks known collectively as the histone code. The chromatin marks at a given promoter determine, in part, whether specific promoters are in an open/active conformation or closed/repressed conformation. Dimethyl-lysine 4 histone H3 (H3K4me2) is a transcription activating chromatin mark at gene promoters, and demethylation of this mark by the lysine-specific demethylase 1 (LSD1), a homologue of polyamine oxidases, may broadly repress gene expression. We now report that novel biguanide and bisguanidine polyamine analogues are potent inhibitors of LSD1. These analogues inhibit LSD1 in human colon carcinoma cells and affect a reexpression of multiple, aberrantly silenced genes important in the development of colon cancer, including members of the secreted frizzle-related proteins (SFRPs) and the GATA family of transcription factors. Furthermore, we demonstrate by chromatin immunoprecipitation analysis that the reexpression is concurrent with increased H3K4me2 and acetyl-H3K9 marks, decreased H3K9me1 and H3K9me2 repressive marks. We thus define important new agents for reversing aberrant repression of gene transcription. PMID- 17463087 TI - Cell proliferation and survival induced by Toll-like receptors is antagonized by type I IFNs. AB - TRIF is an adaptor protein associated with the signaling by Toll-like receptor (TLR)3 and TLR4 for the induction of type I IFNs. Here, we demonstrate a mechanism by which TLR signaling controls cell proliferation and survival. We show that TLR3 and TLR4 can induce cell cycle entry via TRIF, which targets the cell cycle inhibitor p27(kip1) for relocalization, phosphorylation by cyclin/cdk complexes, and proteasome degradation. These events are antagonized by type I IFN induced by the TRIF pathway. Furthermore, in human dendritic cells treated with TLR3, TLR4, or TLR5 ligands, we demonstrate that IFN signaling modulates p27(kip1) degradation and apoptosis, identifying an immunoregulatory "switching" function of type I IFNs. These findings reveal a previously uncharacterized function of TLR signaling in cell proliferation and survival. PMID- 17463088 TI - GATA6 is an astrocytoma tumor suppressor gene identified by gene trapping of mouse glioma model. AB - Malignant astrocytomas are the most common and lethal adult primary brain tumor. Retroviral gene trapping of nontransformed neonatal astrocytes from a glial fibrillary acidic protein (GFAP):(V12)Ha-Ras murine astrocytoma model led to isolation of the transcription factor Gata6. Loss of Gata6 resulted in enhanced proliferation and transformation of astrocytes. Human malignant astrocytoma cell lines, explant xenografts, and operative specimens demonstrated loss of GATA6 expression. Loss-of-function GATA6 mutations with loss of heterozygosity of the GATA6 locus were found in human malignant astrocytoma specimens but not in lower grade astrocytomas or normal adult astrocytes. Knockdown of Gata6 expression in (V12)Ha-Ras or p53-/- astrocytes, but not in parental murine or human astrocytes, led to acceleration of tumorgenesis. Knockin GATA6 expression in human malignant astrocytoma cells reduced their tumorgenic growth with decreased VEGF expression. Collectively, these data demonstrate that GATA6, isolated from a murine astrocytoma model, is a novel tumor suppressor gene that is a direct target of mutations during malignant progression of murine and human astrocytomas. This work also demonstrates the utility of random mutagenesis strategies, such as gene trapping, on murine cancer models toward discovery of novel genetic alterations in corresponding human cancers. PMID- 17463089 TI - Thousands of human mobile element fragments undergo strong purifying selection near developmental genes. AB - At least 5% of the human genome predating the mammalian radiation is thought to have evolved under purifying selection, yet protein-coding and related untranslated exons occupy at most 2% of the genome. Thus, the majority of conserved and, by extension, functional sequence in the human genome seems to be nonexonic. Recent work has highlighted a handful of cases where mobile element insertions have resulted in the introduction of novel conserved nonexonic elements. Here, we present a genome-wide survey of 10,402 constrained nonexonic elements in the human genome that have all been deposited by characterized mobile elements. These repeat instances have been under strong purifying selection since at least the boreoeutherian ancestor (100 Mya). They are most often located in gene deserts and show a strong preference for residing closest to genes involved in development and transcription regulation. In particular, constrained nonexonic elements with clear repetitive origins are located near genes involved in cell adhesion, including all characterized cellular members of the reelin-signaling pathway. Overall, we find that mobile elements have contributed at least 5.5% of all constrained nonexonic elements unique to mammals, suggesting that mobile elements may have played a larger role than previously recognized in shaping and specializing the landscape of gene regulation during mammalian evolution. PMID- 17463090 TI - Selective chemical probe inhibitor of Stat3, identified through structure-based virtual screening, induces antitumor activity. AB - S3I-201 (NSC 74859) is a chemical probe inhibitor of Stat3 activity, which was identified from the National Cancer Institute chemical libraries by using structure-based virtual screening with a computer model of the Stat3 SH2 domain bound to its Stat3 phosphotyrosine peptide derived from the x-ray crystal structure of the Stat3beta homodimer. S3I-201 inhibits Stat3.Stat3 complex formation and Stat3 DNA-binding and transcriptional activities. Furthermore, S3I 201 inhibits growth and induces apoptosis preferentially in tumor cells that contain persistently activated Stat3. Constitutively dimerized and active Stat3C and Stat3 SH2 domain rescue tumor cells from S3I-201-induced apoptosis. Finally, S3I-201 inhibits the expression of the Stat3-regulated genes encoding cyclin D1, Bcl-xL, and survivin and inhibits the growth of human breast tumors in vivo. These findings strongly suggest that the antitumor activity of S3I-201 is mediated in part through inhibition of aberrant Stat3 activation and provide the proof-of-concept for the potential clinical use of Stat3 inhibitors such as S3I 201 in tumors harboring aberrant Stat3. PMID- 17463091 TI - Separate oscillating cell groups in mouse suprachiasmatic nucleus couple photoperiodically to the onset and end of daily activity. AB - The pattern of circadian behavioral rhythms is photoperiod-dependent, highlighted by the conservation of a phase relation between the behavioral rhythm and photoperiod. A model of two separate, but mutually coupled, circadian oscillators has been proposed to explain photoperiodic responses of behavioral rhythm in nocturnal rodents: an evening oscillator, which drives the activity onset and entrains to dusk, and a morning oscillator, which drives the end of activity and entrains to dawn. Continuous measurement of circadian rhythms in clock gene Per1 expression by a bioluminescence reporter enabled us to identify the separate oscillating cell groups in the mouse suprachiasmatic nucleus (SCN), which composed circadian oscillations of different phases and responded to photoperiods differentially. The circadian oscillation in the posterior SCN was phase-locked to the end of activity under three photoperiods examined. On the other hand, the oscillation in the anterior SCN was phase-locked to the onset of activity but showed a bimodal pattern under a long photoperiod [light-dark cycle (LD)18:6]. The bimodality in the anterior SCN reflected two circadian oscillatory cell groups of early and late phases. The anterior oscillation was unimodal under intermediate (LD12:12) and short (LD6:18) photoperiods, which was always phase lagged behind the posterior oscillation when the late phase in LD18:6 was taken. The phase difference was largest in LD18:6 and smallest in LD6:18. These findings indicate that three oscillating cell groups in the SCN constitute regionally specific circadian oscillations, and at least two of them are involved in photoperiodic response of behavioral rhythm. PMID- 17463092 TI - Parasite-intrinsic factors can explain ordered progression of trypanosome antigenic variation. AB - Pathogens often persist during infection because of antigenic variation in which they evade immunity by switching between distinct surface antigen variants. A central question is how ordered appearance of variants, an important determinant of chronicity, is achieved. Theories suggest that it results directly from a complex pattern of transition connectivity between variants or indirectly from effects such as immune cross-reactivity or differential variant growth rates. Using a mathematical model based only on known infection variables, we show that order in trypanosome infections can be explained more parsimoniously by a simpler combination of two key parasite-intrinsic factors: differential activation rates of parasite variant surface glycoprotein (VSG) genes and density-dependent parasite differentiation. The model outcomes concur with empirical evidence that several variants are expressed simultaneously and that parasitaemia peaks correlate with VSG genes within distinct activation probability groups. Our findings provide a possible explanation for the enormity of the recently sequenced VSG silent archive and have important implications for field transmission. PMID- 17463093 TI - From the liver to the brain across the blood-brain barrier. PMID- 17463094 TI - Toxicity from different SOD1 mutants dysregulates the complement system and the neuronal regenerative response in ALS motor neurons. AB - Global, age-dependent changes in gene expression from rodent models of inherited ALS caused by dominant mutations in superoxide-dismutase 1 (SOD1) were identified by using gene arrays and RNAs isolated from purified embryonic and adult motor neurons. Comparison of embryonic motor neurons expressing a dismutase active ALS linked mutant SOD1 with those expressing comparable levels of wild-type SOD1 revealed the absence of mutant-induced mRNA changes. An age-dependent mRNA change that developed presymptomatically in adult motor neurons collected by laser microdissection from mice expressing dismutase active ALS-linked mutants was dysregulation of the d/l-serine biosynthetic pathway, previously linked to both excitotoxic and neurotrophic effects. An unexpected dysregulation common to motor neurons expressing either dismutase active or inactive mutants was induction of neuronally derived components of the classic complement system and the regenerative/injury response. Alteration of these mutant SOD1-induced pathways identified a set of targets for therapies for inherited ALS. PMID- 17463095 TI - A comparative study of simple bone cysts of the jaw and extracranial bones. AB - OBJECTIVES: To improve the interpretation of simple bone cyst (SBC) lesions of the jaw. METHODS: A comparative study of SBC lesions of the jaw and extracranial bones was performed through a literature survey. RESULTS: In extracranial SBC, the cavities were always filled with fluid, and a high recurrence rate was shown through extensive research. Aneurysmal bone cyst (ABC) was included in the differential diagnosis owing to some clinicopathologic similarities. Fluid, gas and blood were found in the cavity in jawbone SBC, and recurrence was believed to be rare. Differential diagnosis was rarely discussed in the literature. CONCLUSIONS: Based on reports, the cavity did not normally contain gas because no air-fluid level was observed on panoramic radiographs and no density/intensity area indicating gas was seen on CT or MRI. A blood-filled cavity should be examined carefully, and the possibility of an ABC should be considered. The recurrence rate needs to be re-estimated because an extensive survey has not been performed to clarify the treatment outcomes of jawbone SBC. PMID- 17463096 TI - Density value means in the evaluation of external apical root resorption: an in vitro study for early detection in orthodontic case simulations. AB - OBJECTIVES: The purpose of this study was to develop an alternative diagnostic tool for the early detection of external apical root resorption (EARR). METHODS: Mandibular incisors (n=36) with and without simulated EARR lesions were used. 18 teeth with facial and proximal windows, each with a range of 2 sizes, were placed in 6 N hydrochloric acid (HCl) baths for 10 min. A sample of the acid solution was analysed for calcium concentration by atomic absorption spectrophotometry. Incisors were imaged at 80 degrees, 90 degrees and 100 degrees under 3 test conditions (bracketed, non-bracketed and with subtraction registration templates (SRTs)). The images were reconstructed and subtracted to determine the accuracy and sensitivity of the method. Quantified histograms for each subtracted image were constructed. RESULTS: At either an angle of 80 degrees or 100 degrees, the bracketed group had the largest mean standard deviation of the subtraction histograms while the SRT group had the smallest. Density values as a function of total calcium removed were plotted indicating a linear relationship between subtraction density units and calcium loss. CONCLUSION: The use of the SRTs was significantly more accurate than the use of the brackets alone for digital subtraction radiography reconstructions. This model shows promise for detecting EARR prior to a noticeable decrease in root length. It may be useful for early detection of resorptive lesions during routine orthodontic treatment. PMID- 17463097 TI - In vitro comparison of conventional film and direct digital imaging in the detection of approximal caries. AB - OBJECTIVES: To compare the diagnostic accuracy of conventional film, unenhanced direct digital and inversion grayscale direct digital imaging in the detection of approximal caries. METHODS: 150 approximal surfaces of extracted permanent molars and premolars were selected for the study on the basis of varying lesion depth. The teeth were radiographed using Ektaspeed Plus film; digital images were made with a Schick CMOS-APS sensor. 7 examiners evaluated 58 randomized images of each modality. Histological sectioning of the teeth was used to verify the presence and extent of decay. RESULTS: No significant difference was found between the diagnostic accuracies of the three imaging modalities (P=0.226). Analysis of the diagnostic accuracy of the three modalities on lesion depth showed no statistically significant interaction; however, the main effect of the lesion depth was significant (P<0.001, eta(2)=0.936). CONCLUSIONS: The overall diagnostic accuracy of the three modalities in the detection of approximal carious lesions was comparable. All three modalities performed poorly in the detection of enamel lesions. PMID- 17463098 TI - Risk of vertebral osteoporosis in post-menopausal women with alterations of the mandible. AB - BACKGROUND: Previous studies have suggested that a thin or eroded cortex of the mandible detected on dental panoramic radiographs is associated with low vertebral bone mineral density (BMD) or osteoporosis. However, those studies did not estimate the multivariate-adjusted risk for low vertebral BMD or osteoporosis associated with alterations of the mandible. METHODS: BMD of the lumbar vertebrae (L2-L4) was compared among quartiles of cortical width and among three cortical shape categories in 450 post-menopausal women (mean age, 57.2 years), adjusted for potential confounders. The odds ratios for low BMD or osteoporosis according to cortical width and shape were also calculated. RESULTS: Significant associations were found between cortical width and shape, and vertebral BMD. The odds ratios for low vertebral BMD associated with the second, third and lowermost quartiles of cortical width were 1.71 (95% confidence interval (CI), 0.96-3.05), 2.30 (95% CI, 1.29-4.11) and 5.43 (95% CI, 2.16-10.71), respectively, compared with the uppermost quartile. The odds ratios for osteoporosis according to cortical width category were similar to those for low BMD. The odds ratios for low BMD associated with mildly to moderately and severely eroded cortices were 3.85 (95% CI, 2.37-6.25) and 7.84 (95% CI, 2.57-23.90), respectively, compared with normal cortex. The odds ratios for osteoporosis associated with mildly to moderately and severely eroded cortices were 4.73 (95% CI, 2.54-8.80) and 14.73 (95% CI, 6.14-35.47), respectively. CONCLUSIONS: Post-menopausal women with alterations of the mandible may have an increased risk for low vertebral BMD or osteoporosis. PMID- 17463099 TI - Identification of post-menopausal women at risk of osteoporosis by trained general dental practitioners using panoramic radiographs. AB - BACKGROUND: An eroded inferior cortex of the mandible on panoramic radiographs may be useful for identifying post-menopausal women with low bone mineral density (BMD), or osteoporosis. The purpose of this study was to assess whether trained general dental practitioners (GDPs) can identify post-menopausal women with undetected low skeletal BMD as well as spinal fractures by panoramic radiographs in their clinics. METHODS: Out of 455 women aged 50 years and older who visited the dental clinics of 22 trained GDPs and had panoramic radiographic assessment for the examination of dental diseases between June and December 2004, 168 post menopausal women were diagnosed as having low skeletal BMD based on cortical erosion findings. Of these women, 39 women aged 50-84 years (mean age (SD, standard deviation), 64.8 (7.4) years) with no previous diagnosis of osteoporosis participated in this study. BMD at the lumbar spine and femoral neck was measured using dual energy X-ray absorptiometry (DXA). Spine fractures were assessed on lateral radiographs obtained at the time of the DXA assessment. RESULTS: Two women (5.1%) had normal BMD (BMD T-score>-1.0), 21 women (53.9%) had osteopenia (BMD T-score of -2.5 to -1.0) and 16 women (41.0%) had osteoporosis (BMD T-score< 2.5). Eight women (20.5%) had fractures at the thoracic spine, lumbar spine, or both. CONCLUSIONS: Our results suggest that a high percentage of post-menopausal women with undetected low skeletal BMD as well as spinal fractures may be identified based on trained GDPs' analyses of their panoramic radiographs. PMID- 17463100 TI - Baseline clinical and radiographic features are associated with long-term (8 years) signs/symptoms for subjects with diseased temporomandibular joint. AB - OBJECTIVES: The purpose of this study was to determine the extent to which baseline clinical and radiographic features were associated with long-term outcomes in patients with temporomandibular joint disorder (TMJD). METHODS: 49 patients with unilateral radiographically proven TMJD were available in this study. Self-reported long-term (mean 96.2 months) outcomes (current joint pain, maximum mouth opening and joint noise) after TMJD treatments were assessed by questionnaire. The impact of multiple initial clinical/radiographic findings (gender, age at first visit, time interval between first visit and questionnaire survey, treatment method, disc displacement, disc morphology, disc mobility, condylar bony change and morphology of the articular eminence) on the long-term outcomes was assessed using stepwise multiple regression and logistic regression analysis. RESULTS: Patient age at the first visit was significantly correlated with current joint pain. Disc mobility and morphology of the articular eminence were significantly correlated with current range of maximal mouth opening. CONCLUSIONS: The results of this study suggest that patients who appeared symptomatic at a younger age or who initially had a fixed disc were the most likely to have recurrent or persisting clinical signs/symptoms of TMJD after 8 years. PMID- 17463101 TI - Radiographic examination of 41 cases of odontogenic myxomas on the basis of conventional radiographs. AB - OBJECTIVE: There are some different radiographic appearances of odontogenic myxoma that have not been reported in the published literature. The purpose of this article is to offer rare views of odontogenic myxomas and analyse the characteristics of the radiographic appearances. MATERIALS AND METHODS: We reviewed the radiographic appearances of 41 cases of odontogenic myxomas on conventional radiographs, recruited from patients who visited the Department of Oral Radiology, West China College of Stomatology, Sichuan University between 1964 and 2005. The diagnoses of the cases were verified by histopathology. RESULTS: On conventional radiographs, odontogenic myxomas presented varying radiographic appearances, which could be divided into six types as follows: Type I-unilocular; Type II-multilocular (including honeycomb, soap bubble and tennis racquet patterns); Type III-involvement of local alveolar bone; Type IV involvement of the maxillary sinus; Type V-osteolytic destruction and Type VI-a mix of osteolytic destruction and osteogenesis. CONCLUSION: Odontogenic myxoma has variable radiographic appearances on conventional radiographs. This classification system helps us better understand the radiographic appearances of odontogenic myxoma on conventional radiographs. PMID- 17463102 TI - Micronucleus test in buccal epithelium cells from patients subjected to panoramic radiography. AB - OBJECTIVES: To evaluate the possible genotoxic effect of radiation exposure for dental diagnostic purposes as measured by the formation of micronuclei. METHODS: The micronucleus test was applied to buccal epithelium cells, which are target cells for dental radiography. Specimens of exfoliated buccal cells were collected from patients subjected to panoramic radiography. Samples were obtained from 32 patients, 12 male and 20 female, aged from 24 years to 73 years, before and 10+/ 2 days after panoramic radiation exposure. RESULTS: No significant increase in the frequency of cells with micronuclei and total number of micronuclei after panoramic tomography was detected. Mean values of buccal cells with micronuclei+/ standard deviation (SD) before and after radiation examination were 2.34+/-1.49% and 2.81+/-1.64%, respectively. A significant correlation between the age of investigated subjects and the initial frequency of micronuclei in buccal cells was observed (r=0.60, P<0.01). CONCLUSION: Panoramic radiographic examination does not induce micronuclei in target buccal epithelium cells. PMID- 17463103 TI - Facial subcutaneous calcinosis and mandibular resorption in systemic sclerosis: a case report. AB - Systemic sclerosis (SSc) is an autoimmune disease characterized by widespread fibrosis of subcutaneous connective tissue, causing serious complications with the involvement of other systems. Tight, firm skin is the main characteristic of SSc, causing extrinsic pressure and the obliteration of the vessels. This then leads to ischaemia and destruction of the underlying bone. This mechanism may also lead to bone resorption of the mandibular body as it affects other bones of the skeleton. To date, only two cases of reported calcinosis in the facial area have been observed. In this report, a rare case of SSc with both subcutaneous calcinosis and mandibular bone resorption is presented. The role of dental practitioners in the diagnosis of the disease with respect to the orofacial symptoms is also discussed. PMID- 17463104 TI - Ultrasonographic and arthrographic diagnoses of synovial chondromatosis. AB - Synovial chondromatosis (SC) is rare in the temporomandibular joint (TMJ). Plain radiographs, CT, MRI and arthrography of the TMJ have been used for the diagnosis of SC. The purpose of this paper is to present ultrasonographic and arthrographic diagnoses of a case of SC of the TMJ. PMID- 17463105 TI - Peripheral ossifying fibroma in the oral cavity: CT and MR findings. AB - We report a case of peripheral ossifying fibroma arising from the anterior oral cavity in a 12-year-old boy. CT and MR scans showed a large exophytic soft tissue mass overlying the anterior hard palate and maxillary alveolar ridge. The tumour revealed peripheral calcification without adjacent bone changes. PMID- 17463106 TI - Peripheral ameloblastoma: a case report. AB - Peripheral ameloblastoma, a rare and unusual variant of odontogenic tumour, comprises about 1% of all ameloblastomas. The extraosseous location is the peculiar feature of this type of tumour, which is otherwise similar to the classical ameloblastoma. An additional case of this infrequent tumour is described on the lingual alveolar mucosa of the right mandibular premolar region in a 60-year-old man. Clinical evidence, microscopic findings, CT imaging and recurrence for this tumour are discussed. PMID- 17463107 TI - The anabolic and catabolic responses in bone repair. AB - The literature on fracture repair has been reviewed. The traditional concepts of delayed and nonunion have been examined in terms of the phased and balanced anabolic and catabolic responses in bone repair. The role of medical manipulation of these inter-related responses in the fracture healing have been considered. PMID- 17463108 TI - The prophylaxis and treatment of heterotopic ossification following lower limb arthroplasty. AB - Heterotopic ossification following joint replacement in the lower limb occurs in 3% to 90% of cases. Higher grades of heterotopic ossification can result in significant limitation of function and can negate the benefits of joint replacement. The understanding of the pathophysiology of this condition has improved in recent years. It would appear to be related to a combination of systemic and local factors, including over-expression of bone morphogenetic protein-4. There is currently little evidence to support the routine use of prophylaxis for heterotopic ossification in arthroplasty patients, but prophylaxis is recommended by some for high-risk patients. Radiotherapy given as one dose of 7 Gy to 8 Gy, either pre-operatively (< four hours before) or post operatively (within 72 hours of surgery), appears to be more effective than indometacin therapy (75 mg daily for six weeks). In cases of prophylaxis against recurrent heterotopic ossification following excision, recent work has suggested that a combination of radiotherapy and indometacin is effective. Advances in our understanding of this condition may permit the development of newer, safer treatment modalities. PMID- 17463109 TI - Chromium and cobalt ion release following the Durom high carbon content, forged metal-on-metal surface replacement of the hip. AB - We evaluated the concentrations of chromium and cobalt ions in blood after metal on-metal surface replacement arthroplasty using a wrought-forged, high carbon content chromium-cobalt alloy implant in 64 patients. At one year, mean whole blood ion levels were 1.61 microg/L (0.4 to 5.5) for chromium and 0.67 microg/L (0.23 to 2.09) for cobalt. The pre-operative ion levels, component size, female gender and the inclination of the acetabular component were inversely proportional to the values of chromium and/or cobalt ions at one year postoperatively. Other factors, such as age and level of activity, did not correlate with the levels of metal ions. We found that the levels of the ions in the serum were 1.39 and 1.37 times higher for chromium and cobalt respectively than those in the whole blood. The levels of metal ions obtained may be specific to the hip resurfacing implant and reflect its manufacturing process. PMID- 17463110 TI - Long-term survivorship of the Charnley Elite Plus femoral component in young patients. AB - We studied prospectively the long-term results of the Charnley Elite-Plus femoral stem in 184 consecutive young patients (194 hips). There were 130 men and 54 women with a mean age of 49.1 years (21 to 60). The predominant diagnosis was osteonecrosis of the femoral head (63.6%, 117 patients). Clinical and radiological evaluation was undertaken at each follow-up. The mean follow-up was 11.2 years (10 to 12). The mean pre-operative Harris hip score was 43.4 (12 to 49) which improved to 91 (59 to 100) at the final follow-up. The survival of the femoral stem at 12 years was 99% with revision as the end-point. The mean annual linear wear of the polyethylene liner was 0.17 mm (0.13 to 0.22). The prevalence of acetabular osteolysis was 10.8% (21 hips) and osteolysis of the calcar femorale 12.9% (25 hips). A third-generation cementing technique, accurate alignment of the stem and the use of a 22 mm zirconia head were important factors in the prevention of aseptic loosening of the Elite Plus femoral stem in these high-risk young patients. PMID- 17463111 TI - Mid-term results of cementless total hip replacement using a ceramic-on-ceramic bearing with and without computer navigation. AB - We have developed a CT-based navigation system using infrared light-emitting diode markers and an optical camera. We used this system to perform cementless total hip replacement using a ceramic-on-ceramic bearing couple in 53 patients (60 hips) between 1998 and 2001. We reviewed 52 patients (59 hips) at a mean of six years (5 to 8) postoperatively. The mid-term results of total hip replacement using navigation were compared with those of 91 patients (111 hips) who underwent this procedure using the same implants, during the same period, without navigation. There were no significant differences in age, gender, diagnosis, height, weight, body mass index, or pre-operative clinical score between the two groups. The operation time was significantly longer where navigation was used, but there was no significant difference in blood loss or navigation-related complications. With navigation, the acetabular components were placed within the safe zone defined by Lewinnek, while without, 31 of the 111 components were placed outside this zone. There was no significant difference in the Merle d'Aubigne and Postel hip score at the final follow-up. However, hips treated without navigation had a higher rate of dislocation. Revision was performed in two cases undertaken without navigation, one for aseptic acetabular loosening and one for fracture of a ceramic liner, both of which showed evidence of neck impingement on the liner. A further five cases undertaken without navigation showed erosion of the posterior aspect of the neck of the femoral component on the lateral radiographs. These seven impingement-related mechanical problems correlated with malorientation of the acetabular component. There were no such mechanical problems in the navigated group. We conclude that CT-based navigation increased the precision of orientation of the acetabular component and control of limb length in total hip replacement, without navigation-related complications. It also reduced the rate of dislocation and mechanical problems related to impingement. PMID- 17463112 TI - Mortality and mobility after hip fracture in Japan: a ten-year follow-up. AB - We studied prospectively the change over ten years in mortality, walking ability and place of residence after a hip fracture in 753 patients in Japan. We compared the deaths observed in these patients with those expected in the general population, matched for age, gender and calender year at the time of fracture. The survival rate decreased dramatically for two years after the event and the mortality risk remained higher for ten years. This risk was approximately double that of the general population, even at ten years after fracture. The risk was higher, and remained so for longer, in younger rather than in older patients. The proportion of patients who were able to walk outdoors alone, with or without an assistive device, was 68% (514) before fracture. This decreased to 56% (340) by one year after and remained stable at approximately 63% (125) until ten years. The proportion of patients living in their own home was 84% (629) before fracture, 81% (491) one year later, and then remained stable at approximately 86% (171) until ten years after the event. PMID- 17463113 TI - Clinical outcome and rate of complications after primary total knee replacement performed with quadriceps-sparing or standard arthrotomy. AB - We performed a prospective, randomised study to compare the results and rates of complications of primary total knee replacement performed using a quadriceps sparing technique or a standard arthrotomy in 120 patients who had bilateral total knee replacements carried out under the same anaesthetic. The clinical results, pain scales, surgical and hospital data, post-operative complications and radiological results were compared. No significant differences were found between the two groups with respect to the blood loss, knee score, function score, pain scale, range of movement or radiological findings. In contrast, the operating time (p = 0.0001) and the tourniquet time (p < 0.0001) were significantly longer in the quadriceps-sparing group, as was the rate of complications (p = 0.0468). We therefore recommend the use of a standard arthrotomy with the shortest possible skin incision for total knee replacement. PMID- 17463114 TI - Alignment and orientation of the components in total knee replacement with and without navigation support: a prospective, randomised study. AB - Bilateral sequential total knee replacement was carried out under one anaesthetic in 100 patients. One knee was replaced using a CT-free computer-assisted navigation system and the other conventionally without navigation. The two methods were compared for accuracy of orientation and alignment of the components. There were 85 women and 15 men with a mean age of 67.6 years (54 to 83). Radiological and CT imaging was carried out to determine the alignment of the components. The mean follow-up was 2.3 years (2 to 3). The operating and tourniquet times were significantly longer in the navigation group (p < 0.001). There were no significant pre- or post-operative differences between the knee scores of the two groups (p = 0.288 and p = 0.429, respectively). The results of imaging and the number of outliers for all radiological parameters were not statistically different (p = 0.109 to p = 0.920). In this series computer assisted navigated total knee replacement did not result in more accurate orientation and alignment of the components than that achieved by conventional total knee replacement. PMID- 17463115 TI - Computer navigation versus conventional total knee replacement: no difference in functional results at two years. AB - We previously compared the component alignment in total knee replacement using a computer-navigated technique with a conventional jig-based method. We randomly allocated 71 patients to undergo either computer-navigated or conventional replacement. An improved alignment was seen in the computer-navigated group. The patients were then followed up post-operatively for two years, using the Knee Society score, the Short Form-36 health survey, the Western Ontario and McMaster Universities osteoarthritis index, the Bartlett Patellar pain questionnaire and the Oxford knee score, to assess functional outcome. At two years post operatively 60 patients were available for assessment, 30 in each group and 62 patients completed a postal survey. No patient in either group had undergone revision. All variables were analysed for differences between the groups either by Student's t-test or the Mann-Whitney U test. Differences between the two groups did not reach significance for any of the outcome measures at any time point. At two years postoperatively, the frequency of mild to severe anterior pain was not significantly different (p = 0.818), varying between 44% (14) for the computer-navigated group, and 47% (14) for the conventionally-replaced group. The Bartlett Patellar score and the Oxford knee score were also not significantly different (t-test p = 0.161 and p = 0.607, respectively). The clinical outcome of the patients with a computer-navigated knee replacement appears to be no different to that of a more conventional jig-based technique at two years post operatively, despite the better alignment achieved with computer-navigated surgery. PMID- 17463116 TI - Embolic phenomena during computer-assisted and conventional total knee replacement. AB - Systemic emboli released during total knee replacement have been implicated as a cause of peri-operative morbidity and neurological dysfunction. We undertook a prospective, double-blind, randomised study to compare the cardiac embolic load sustained during computer-assisted and conventional, intramedullary-aligned, total knee replacement, as measured by transoesophageal echocardiography. There were 26 consecutive procedures performed by a single surgeon at a single hospital. The embolic load was scored using the modified Mayo grading system for echogenic emboli. Fourteen patients undergoing computer-assisted total knee replacement had a mean embolic score of 4.89 (3 to 7) and 12 undergoing conventional total knee replacement had a mean embolic score of 6.15 (4 to 8) on release of the tourniquet. Comparison of the groups using a two-tailed t-test confirmed a highly significant difference (p = 0.004). This study demonstrates that computer-assisted knee replacement results in the release of significantly fewer systemic emboli than the conventional procedure using intramedullary alignment. PMID- 17463117 TI - Early results of the Acclaim elbow replacement. AB - The Acclaim total elbow replacement is a modular system which allows implantation in both unlinked and linked modes. The results of the use of this implant in primary total elbow replacement in 36 patients, operated on between July 2000 and August 2002, are presented at a mean follow-up of 36 months (24 to 49). Only one patient did not have good relief of pain, but all had improved movement and function. No implant showed clinical or radiological loosening, although one had a lucent area in three of seven humeral zones. The short-term results of the Acclaim total elbow replacement are encouraging. However, 11 patients (30.5%) suffered an intra-operative fracture of the humeral condyle. This did not affect the outcome, or the requirement for further surgery, except in one case where the fracture failed to unite. This problem has hopefully been addressed by redesigning the humeral resection guide. Other complications included three cases of ulnar neuropathy (8.3%) and one of deep infection (2.8%). PMID- 17463118 TI - A three-dimensional quantitative analysis of carpal deformity in rheumatoid wrists. AB - We have measured the three-dimensional patterns of carpal deformity in 20 wrists in 20 rheumatoid patients in which the carpal bones were shifted ulnarwards on plain radiography. Three-dimensional bone models of the carpus and radius were created by computerised tomography with the wrist in the neutral position. The location of the centroids and rotational angle of each carpal bone relative to the radius were calculated and compared with those of ten normal wrists. In the radiocarpal joint, the proximal row was flexed and the centroids of all carpal bones translocated in an ulnar, proximal and volar direction with loss of congruity. In the midcarpal joint, the distal row was extended and congruity generally well preserved. These findings may facilitate more positive use of radiocarpal fusion alone for the deformed rheumatoid wrist. PMID- 17463119 TI - Balloon kyphoplasty as a single or as an adjunct procedure for the management of symptomatic vertebral haemangiomas. AB - Vertebral haemangiomas are usually asymptomatic and discovered fortuitously during imaging. A small proportion may develop variable degrees of pain and neurological deficit. We prospectively studied six patients who underwent eight surgical procedures on 11 vertebral bodies. There were 11 balloon kyphoplasties, six lumbar and five thoracic. The mean follow-up was 22.3 months (12 to 36). The indications for operation were pain in four patients, severe back pain with Frankel grade C paraplegia from cord compression caused by soft-tissue extension from a thoracic vertebral haemangioma in one patient, and acute bleeding causing Frankel grade B paraplegia from an asymptomatic vascular haemangioma in one patient. In four patients the exhibited aggressive vascular features, and two showed lipomatous, non-aggressive, characteristics. One patient who underwent a unilateral balloon kyphoplasty developed a recurrence of symptoms from the non treated side of the vertebral body which was managed by a further similar procedure. Balloon kyphoplasty was carried out successfully and safely in all patients; four became asymptomatic and two showed considerable improvement. Neurological recovery occurred in all cases but bleeding was greater than normal. To avoid recurrence, complete obliteration of the lesion with bone cement is indicated. For acute bleeding balloon kyphoplasty should be combined with emergency decompressive laminectomy. For intraspinal extension with serious neurological deficit, a combination of balloon kyphoplasty with intralesional alcohol injection is effective. PMID- 17463120 TI - Two-level reconstruction of comminuted posterior-wall fractures of the acetabulum. AB - Our aim was to evaluate the efficacy of a two-level reconstruction technique using subchondral miniscrews for the stabilisation of comminuted posterior-wall marginal acetabular fragments before applying lag screws and a buttress plate to the main overlying posterior fragment. Between 1995 and 2003, 29 consecutive patients with acute comminuted displaced posterior-wall fractures of the acetabulum were treated operatively using this technique. The quality of reduction measured from three standard plain radiographs was graded as anatomical in all 29 hips. The clinical outcome at a mean follow-up of 35 months (24 to 90) was considered to be excellent in five patients (17%), very good in 16 (55%), good in six (21%) and poor in two (7%). The use of the two-level reconstruction technique appears to provide stable fixation and is associated with favourable results in terms of the incidence of post-traumatic osteoarthritis and the clinical outcome. However, poor results may occur in patients over the age of 55 years. PMID- 17463121 TI - Fractures of the distal humeral articular surface. AB - Fractures of the distal humeral articular surface which do not involve the medial and lateral columns are often more extensive than is apparent from plain radiographs. This retrospective study describes the epidemiology of this injury using modern classification systems and compares pre-operative radiography with operative findings. The study group included 79 patients with a mean age of 47 years (13 to 91). The annual incidence was 1.5 per 100,000 population, and was highest in women over the age of 60. The majority of the fractures (59; 75%) were sustained in falls from standing height. Young males tended to sustain more high energy injuries with more complex fracture patterns. In 24% of cases (19) there was a concomitant radial head fracture. Classification from plain radiographs often underestimates the true extent of the injury and computed tomography may be of benefit in pre-operative planning, especially in those over 60 years of age. PMID- 17463122 TI - Reverse shoulder arthroplasty for the treatment of three- and four-part fractures of the proximal humerus in the elderly: a prospective review of 43 cases with a short-term follow-up. AB - We used an inverted shoulder arthroplasty in 43 consecutive patients with a mean age of 78 years (65 to 97) who had sustained a three- or four-part fracture of the upper humerus. All except two were reviewed with a mean follow-up of 22 months (6 to 58). The clinical outcome was satisfactory with a mean active anterior elevation of 97 degrees (35 degrees to 160 degrees ) and a mean active external rotation in abduction of 30 degrees (0 degrees to 80 degrees ). The mean Constant and the mean modified Constant scores were respectively 44 (16 to 69) and 66% (25% to 97%). Complications included three patients with reflex sympathetic dystrophy, five with neurological complications, most of which resolved, and one with an anterior dislocation. Radiography showed peri prosthetic calcification in 36 patients (90%), displacement of the tuberosities in 19 (53%) and a scapular notch in ten (25%). Compared with conventional hemiarthroplasty, satisfactory mobility was obtained despite frequent migration of the tuberosities. However, long-term results are required before reverse shoulder arthroplasty can be recommended as a routine procedure in complex fractures of the upper humerus in the elderly. PMID- 17463123 TI - Endoprosthetic replacement of the distal femur for bone tumours: long-term results. AB - We investigated whether improvements in design have altered the outcome for patients undergoing endoprosthetic replacement of the distal femur after resection of a tumour. Survival of the implant and 'servicing' procedures have been documented using a prospective database, review of the design of the implant and case records. In total, 335 patients underwent a distal femoral replacement, 162 having a fixed-hinge design and 173 a rotating-hinge. The median age of the patients was 24 years (interquartile range 17 to 48). A total of 192 patients remained alive with a mean follow-up of 12 years (5 to 30). The risk of revision for any reason was 17% at five years, 33% at ten years and 58% at 20 years. Aseptic loosening was the main reason for revision of the fixed-hinge knees while infection and fracture of the stem were the most common for the rotating-hinge implant. The risk of revision for aseptic loosening was 35% at ten years with the fixed-hinge knee, which has, however, been replaced by the rotating-hinge knee with a hydroxyapatite collar. The overall risk of revision for any reason fell by 52% when the rotating-hinge implant was used. Improvements in the design of distal femoral endoprostheses have significantly decreased the need for revision operations, but infection remains a serious problem. We believe that a cemented, rotating-hinge prosthesis with a hydroxyapatite collar offers the best chance of long-term survival of the prosthesis. PMID- 17463124 TI - Reverse V osteotomy of the distal humerus for the correction of cubitus varus. AB - We reviewed 22 children with cubitus varus who had been treated by a reverse V osteotomy and fixation by cross-pinning and wiring. The mean pre-operative humeral-elbow-wrist angle was -16.9 degrees (-25 degrees to +9 degrees ) and at the latest follow-up it was +7.3 degrees (-2 degrees to +14 degrees ). No child had a lateral prominence greater than 5 mm after correction. An excellent result was achieved in 20 children and a good result in two. We believe that this osteotomy has the advantages of better inherent stability, the avoidance of a prominent lateral condyle after correction and firm fixation allowing early movement. PMID- 17463125 TI - The effect of tourniquet padding on the efficiency of tourniquets of the upper limb. AB - We report the effect of padding on the efficiency of the pneumatic tourniquet for the upper limb. Varying thicknesses of two commercially-available types of orthopaedic padding (Cellona and Velband) were applied to the arms of 20 volunteers, with three pressure transducers placed directly beneath the padding. A tourniquet was positioned over the padding and inflated to 220 mmHg. Significant reductions in the transmitted pressure were recorded from the transducers with both padding materials. With eight layers of padding, reductions in pressure of 13% (1% to 26%) and 18% (7% to 35%) were seen with Cellona and Velband, respectively. The reduction in pressure with Velband padding correlated with increasing arm circumference (Pearson's correlation coefficient 0.711, p < 0.001). Studies to date have examined how arm circumference affects the required tourniquet inflation pressure. Our study is the first to investigate the effect of the padding and the findings suggest that using more than two layers results in a significant reduction in the transmitted pressure. PMID- 17463126 TI - Surface damage to an Oxinium femoral head prosthesis after dislocation. AB - During open reduction of an irreducible anterior dislocation of a total hip replacement with an Oxinium femoral head, it was observed that the head had been significantly damaged. Gross and scanning electron microscopic examination revealed cracking, gouging, and delamination of the surface. Because of the risk which this poses for damaging the polyethylene acetabular liner, it is strongly recommended that patients with this type of prosthetic head be carefully monitored after a dislocation. PMID- 17463127 TI - Metal ion levels in a triathlete with a metal-on-metal resurfacing arthroplasty of the hip. AB - A prospective study of serum and urinary ion levels was undertaken in a triathlete who had undergone a metal-on-metal resurfacing arthroplasty of the hip four years previously. The one month study period included the final two weeks of training, the day of the triathlon, and the two weeks immediately post-race. Serum cobalt and chromium levels did not vary significantly throughout this period, including levels recorded on the day after the 11-hour triathlon. Urinary excretion of chromium increased immediately after the race and had returned to pre-race levels six days later. The clinical implications are discussed. PMID- 17463128 TI - Osteosarcoma metastasising to the duodenum and pancreas. AB - The incidence of metastatic osteosarcoma is increasing because of improved results following multi-agent chemotherapy and resection of the primary tumour. Metastases occur most commonly in the lungs, whereas bowel metastases are rare. We describe a 25-year-old female who presented with melaena six years after successful resection of an osteosarcoma of her right femur, and one year after resection of a solitary pulmonary metastasis. Imaging revealed a lesion arising within both the duodenum and the pancreas for which a Whipple's pancreatoduodenectomy was carried out, achieving complete resection. Histological examination confirmed the diagnosis of metastatic osteosarcoma. We believe this is only the second such case reported. At 11 months post-operatively she had no detectable disease. Although rare, osteosarcoma can metastasise to the intestine. The surgeon must be aware of this complication, and that bowel metastases are potentially resectable. PMID- 17463129 TI - The early history of tendo Achillis and its rupture. PMID- 17463130 TI - The initial stability and contact mechanics of a press-fit resurfacing arthroplasty of the hip. AB - Finite element analysis was used to examine the initial stability after hip resurfacing and the effect of the procedure on the contact mechanics at the articulating surfaces. Models were created with the components positioned anatomically and loaded physiologically through major muscle forces. Total micromovement of less than 10 mum was predicted for the press-fit acetabular components models, much below the 50 mum limit required to encourage osseointegration. Relatively high compressive acetabular and contact stresses were observed in these models. The press-fit procedure showed a moderate influence on the contact mechanics at the bearing surfaces, but produced marked deformation of the acetabular components. No edge contact was predicted for the acetabular components studied. It is concluded that the frictional compressive stresses generated by the 1 mm to 2 mm interference-fit acetabular components, together with the minimal micromovement, would provide adequate stability for the implant, at least in the immediate post-operative situation. PMID- 17463131 TI - The accuracy of image-free computer navigation in the placement of the femoral component of the Birmingham Hip Resurfacing: a cadaver study. AB - A cadaver study using six pairs of lower limbs was conducted to investigate the accuracy of computer navigation and standard instrumentation for the placement of the Birmingham Hip Resurfacing femoral component. The aim was to place all the femoral components with a stem-shaft angle of 135 degrees . The mean stem-shaft angle obtained in the standard instrumentation group was 127.7 degrees (120 degrees to 132 degrees ), compared with 133.3 degrees (131 degrees to 139 degrees ) in the computer navigation group (p = 0.03). The scatter obtained with computer assisted navigation was approximately half that found using the conventional jig. Computer navigation was more accurate and more consistent in its placement of the femoral component than standard instrumentation. We suggest that image-free computer-assisted navigation may have an application in aligning the femoral component during hip resurfacing. PMID- 17463132 TI - The role of the Pirani scoring system in the management of club foot by the Ponseti method. PMID- 17463133 TI - A score for predicting salvage and outcome in Gustilo type-IIIA and type-IIIB open tibial fractures. PMID- 17463134 TI - Rectal cancer: MR imaging before neoadjuvant chemotherapy and radiation therapy for prediction of tumor-free circumferential resection margins and long-term survival. AB - PURPOSE: To retrospectively evaluate the prognostic importance of involvement of the circumferential resection margin predicted by using magnetic resonance (MR) imaging before neoadjuvant treatment in patients with rectal cancer. MATERIALS AND METHODS: The local institutional review board approved the retrospective analysis of the data and waived informed consent. Sixty-eight patients (52 men, 16 women; mean age +/- standard deviation, 58.9 years +/- 9.4) with cT3 NX M0 tumors were included. T2-weighted MR images were analyzed in consensus by two radiologists with respect to the shortest distance between the outermost parts of the tumor to the adjacent mesorectal fascia (as the potential circumferential resection margin in total mesorectal excision). Histopathologic and follow-up data were available for all patients (mean follow-up time, 54 months; range, 31 77 months). To compare local recurrence and survival rates, the population was divided into three groups categorized according to the minimum distance of the tumor to the mesorectal fascia (group 1, 1 to 5 mm; group 3, >5 mm). Univariate Cox and multivariate proportional hazard regression models were used to test the prognostic importance of clinical, histopathologic regression, and histopathologic tumor parameters. RESULTS: MR imaging led to accurate prediction of a histologically involved circumferential resection margin (sensitivity, 100%; specificity, 88%). The rates for local recurrence (group 1, 33%; group 2, 5%; group 3, 6%; P<.02) and 5-year overall survival (group 1, 39%; group 2, 70%; group 3, 90%; P<.001) differed significantly among the predefined groups. The distance to the mesorectal fascia was an independent prognostic parameter in multivariate analysis (P<.001), and histopathologic response to treatment provided no additional information. CONCLUSION: Prediction of the tumor free circumferential resection margin assessed with MR imaging before initiation of neoadjuvant chemotherapy and radiation therapy proved to be a prognostic factor in rectal cancer. PMID- 17463135 TI - Functional MR imaging during hypercapnia and hyperoxia: noninvasive tool for monitoring changes in liver perfusion and hemodynamics in a rat model. AB - PURPOSE: To prospectively assess functional magnetic resonance (MR) imaging during hypercapnia and hyperoxia for monitoring changes in liver perfusion and hemodynamics in rats. MATERIALS AND METHODS: All experiments were performed with approval of an animal care and use committee. Functional T2*-weighted gradient echo MR images of the rat liver were acquired during hyperoxia and graded hypercapnia (n=24). Additional images were acquired during portal vein ligation (n=4), induced hypovolemia (n=5), and 70% hepatectomy (n=5). Hypercapnic effects were confirmed with Doppler ultrasonography and with gadopentetate dimeglumine. Differences between groups were analyzed by using Wilcoxon rank sum test, except for the graded hypercapnia, for which one-way analysis of variance was used. RESULTS: Liver signal intensity (SI) increased due to hyperoxia; the percentage change in SI was seven times greater than that in muscle tissue; this reflects higher vascularity of the liver. Liver SI decreased due to hypercapnia; the percentage change in SI was negative in the liver but positive in the muscle (P<.001). Induced hypovolemia resulted in considerable decreases in functional MR imaging response; this reflects lower liver perfusion. Clinical applicability of the functional MR imaging method was proved by monitoring changes in liver perfusion that resulted from liver resection. CONCLUSION: In the liver, the magnitude of the percentage change in SI induced by hypercapnia and hyperoxia reflects changes in total blood volume; whereas percentage change in SI values induced by hypercapnia from a negative to a positive value reflects relative changes in portal-to-arterial blood flow ratio. PMID- 17463136 TI - Thymic hyperplasia and thymus gland tumors: differentiation with chemical shift MR imaging. AB - PURPOSE: To prospectively evaluate chemical shift magnetic resonance (MR) imaging for differentiating thymic hyperplasia from tumors of the thymus gland. MATERIALS AND METHODS: The institutional review board approved this study; informed consent was obtained and patient confidentiality was protected. The authors assessed 41 patients (17 male, 24 female; age range, 16-78 years) in whom thymic lesions were seen at chest computed tomography. Patients were assigned to a hyperplasia group (n=23) (18 patients with hyperplastic thymus associated with Graves disease and five with rebound thymic hyperplasia) and a tumor group (n=18) (seven patients with thymomas, four with invasive thymomas, five with thymic cancers, and two with malignant lymphomas). T2-weighted fast spin-echo and T1-weighted in-phase and opposed-phase MR images were obtained in all patients and visually assessed. A chemical shift ratio (CSR), determined by comparing the signal intensity of the thymus gland with that of the paraspinal muscle, was calculated for quantitative analysis. Mean CSRs for the patient groups and subgroups were analyzed by using Welch t and Newman-Keuls tests. P<.05 indicated a significant difference. RESULTS: The thymus gland had homogeneous signal intensity in all 23 patients in the hyperplasia group and in 12 of the 18 patients in the tumor group. The mean CSR (+/- standard deviation) was 0.614 +/- 0.130 in the hyperplasia group and 1.026 +/- 0.039 in the tumor group. Mean CSRs in the patients with a hyperplastic thymus and Graves disease, rebound thymic hyperplasia, thymoma, invasive thymoma, thymic cancer, and malignant lymphoma were 0.594 +/- 0.120, 0.688 +/- 0.154, 1.033 +/- 0.043, 1.036 +/- 0.040, 1.020 +/- 0.044, and 0.997 +/- 0.010, respectively. The difference in CSR between the hyperplasia and tumor groups was significant (P<.001). Mean CSRs in the hyperplasia subgroups were lower than those in the tumor subgroups (P<.001). All hyperplasia group patients had an apparent decrease in thymus gland signal intensity at chemical shift MR imaging; no tumor group patients had a decrease in thymus gland signal intensity. CONCLUSION: Chemical shift MR imaging can be used to differentiate thymic hyperplasia from thymic tumors. PMID- 17463137 TI - Influence of needle tip position on injectate spread in 406 interlaminar lumbar epidural steroid injections. AB - PURPOSE: To retrospectively evaluate flow patterns of interlaminar lumbar epidural steroid injections and compare these patterns to needle tip position. MATERIALS AND METHODS: This study was approved by the institutional review board and complied with HIPAA guidelines. The requirement for informed consent was waived. Fluoroscopic images from 701 consecutive lumbar epidural steroid injections in 485 patients were reviewed. Vertebral level, needle placement, and injectate flow patterns were determined from procedural images. History of previous spinal operations was determined from procedural records. A subset of patients with three or more injections at the same site was also examined. Patterns in the data were evaluated with chi(2) tests and t tests. RESULTS: Images from 406 lumbar epidural steroid injections, one per patient (214 women, 192 men; mean age, 59.8 years), were used for final analysis. Midline injections were less likely to result in unilateral flow than a more lateral approach (P>.001). Flow within only the posterior epidural space was associated with unilateral flow (P=.001). Patients with previous spinal operations were more likely to have cephalad or caudad flow of less than one vertebral level than patients without a history of spinal operations (P<.001). Flow varied considerably within a series of injections in a single patient. CONCLUSION: Epidural injectate flow is highly variable, both among patients and between injections in a single patient. Fluoroscopic monitoring and administration of contrast material mixed with medication, allowing visualization of the full extent of medication flow, are essential to ensure adequate coverage of the target area. PMID- 17463138 TI - Effects of CT irradiation on implantable cardiac rhythm management devices. AB - PURPOSE: To prospectively measure the response of a variety of models of implantable cardiac rhythm management devices (ICRMDs) to the radiation delivered by computed tomography (CT), for both maximum and typical dose levels. MATERIALS AND METHODS: Twenty-one ICRMDs (13 pacemakers, eight cardioverter-defibrillators) manufactured by Medtronic (Minneapolis, Minn) were exposed to ionizing radiation from CT systems in both spiral and dynamic acquisition modes at maximum and typical dose levels. Devices were monitored during exposure to check for any operational abnormalities and were interrogated after exposure to check for any residual abnormalities. Total radiation dose and peak dose rate were measured, and the volume CT dose index was recorded. RESULTS: Oversensing was observed in 20 of 21 devices at maximum doses and in 17 of 20 devices at typical doses. Oversensing most often manifested as inhibition, although it occasionally manifested as tracking or safety pacing. Two devices inhibited for more than 4 seconds in spiral mode at clinical dose levels. Oversensing was transient and ceased as soon as the device stopped moving through the x-ray beam or the beam was turned off. The partial electrical reset (PER) safety feature was activated in two models, InSync 8040 and Thera DR. With the exception of PER, programming was not altered. Effects occurred only if the x-ray beam passed directly over the ICRMD. CONCLUSION: CT irradiation at typical clinical doses results in oversensing of ICRMDs in the majority of devices tested, although the identified effects were predominantly transient. PMID- 17463139 TI - Effects of parenteral fish-oil emulsion (Omegaven) on cutaneous wound healing in rats treated with dexamethasone. AB - BACKGROUND: The aim was to assess wound healing when parenteral fish-oil emulsion is given to rats receiving dexamethasone. METHODS: For 5 days after skin wounding, group S (control; n = 7) received saline 1 mL/kg intraperitoneal (IP); group D (n = 7), dexamethasone 0.2 mg/kg IP; and group DO (n = 9), dexamethasone 0.2 mg/kg IP plus 1 mL/kg Omegaven (Fresenius Kabi, Austria). Wound specimens were assessed for hydroxyproline level, wound depth, histology (epidermal/dermal regeneration, granulation tissue thickness, and angiogenesis), and expression of transforming growth factor-beta (TGF-beta) and platelet-derived growth factor-AA (PDGF-AA). RESULTS: Compared with D and DO specimens, controls had higher hydroxyproline (p < .01), deeper wounds (p < .05), and better histologic scores (p < .01 angiogenesis; others p < .05). There were no significant differences between the group D and DO means for hydroxyproline level, wound depth, or histologic scores (p > .05 for all). Controls had higher TGF-beta expression scores than the other groups (p < .01 for both) and a higher PDGF-AA expression score than group DO (p < .01). Groups D and DO had statistically similar TGF-beta scores, but group D had a higher PDGF-AA score (2.71 +/- 0.75 vs 1.55 +/- 0.72, respectively; p < .05). CONCLUSIONS: According to the parameters we studied, adding parenteral omega-3 and omega-6 fatty acids to the nutrition regimen of rats treated with dexamethasone does not seem to have adverse effects on wound healing, and effects on wound healing may not need to be considered when determining if these agents should be supplemented in nutrition support regimens. PMID- 17463140 TI - A short-term long-chain triglycerides infusion has no influence on immune function of adult patients undergoing gastrointestinal surgery. AB - BACKGROUND: Parenteral nutrition (PN) support containing long-chain triglycerides (LCT) plays a critical supportive role in surgical patients' management. This study aims to investigate the effects of intravenous (IV) LCT emulsion on human immune function in adult patients receiving a gastrointestinal surgical procedure. METHODS: Sixty adult patients were randomly assigned either to the LCT treatment group (n = 32) or to the control group (n = 28). After an abdominal operation, the subjects received PN treatment with or without LCT for 5 days. Neutrophil, peripheral blood mononuclear cell (PBMC), lymphocyte and CD4/CD8, serum immunoglobulin A (IgA), IgG, IgM, complement C3 and C4, interleukin (IL)-2, IL-4, IL-10, IL-12, tumor necrosis factor (TNF)-alpha, and interferon (IFN)-gamma were measured and statistically analyzed. RESULTS: The LCT and control groups did not differ significantly at entry in terms of general features. Except for a significant increase of neutrophil number at 24 hours after the surgery in both groups (p < .01), all parameters representing the patients' immune function had no significant difference between the LCT and the control groups with respect to neutrophil and PBMC count, lymphocyte, CD4/CD8, serum IgA, IgG, IgM, complement C3, C4, IL-2, IL-4, IL-10, IL-12, TNF-alpha, and IFN-gamma (p > .05, respectively) 24 hours before the operation, and 24 hours and 120 hours after the operation. CONCLUSIONS: The regimens of LCT administration may have diverse effects on human immune function in different patient populations. However, LCT emulsion at an appropriate dose and infusion speed does not alter human immune function of adult patients undergoing moderate gastrointestinal surgery. PMID- 17463141 TI - Glycine does not add to the beneficial effects of perioperative oral immune enhancing nutrition supplements in high-risk cardiac surgery patients. AB - BACKGROUND: Elderly patients and patients with a poor cardiac function have increased morbidity rates when undergoing cardiac surgery. The aim of this study was to determine whether addition of glycine to a standard preoperative oral immune-enhancing nutrition supplement (OIENS) improves outcome. Glycine-enriched OIENS was compared with 2 formulas: standard OIENS and control. METHODS: In this double-blind, 3-armed study, patients scheduled to undergo cardiac surgery with the use of extracorporeal circulation received either the glycine-enriched OIENS (OIENS + glyc, n = 24), standard OIENS (OIENS, n = 25), or control formula (Control, n = 25) for minimally 5 preoperative days. Patients were included if they were aged 70 years or older, had a compromised left ventricular function, or were planned for mitral valve surgery. Main outcome measures were postoperative infectious morbidity, organ function, and postoperative recovery. RESULTS: Infectious morbidity was significantly lower in both treatment groups compared with the control group (p = .02). An infection was diagnosed in 5 and 4 patients in the OIENS + glyc and OIENS groups, respectively, and in 12 control patients. Less supportive therapy was necessary to stabilize circulation in both treatment groups compared with the control group. Median length of hospital stay was 7.0, 6.5, and 8.0 days in the OIENS + glyc, OIENS, and control groups, respectively. Inflammatory responses, as measured by systemic levels of proinflammatory cytokines and surface markers on polymorphonuclear cells, were comparable for all groups. CONCLUSIONS: Preoperative OIENS reduces postoperative infectious morbidity and results in a more stable circulation; the addition of glycine does not result in any beneficial effect over standard OIENS. PMID- 17463142 TI - An antioxidative nutrient-rich enteral diet attenuates lethal activity and oxidative stress induced by lipopolysaccharide in mice. AB - BACKGROUND: Oxidative stress is related to various diseases, such as diabetes, cancer, inflammatory disease, and arteriosclerosis. The aim of this study is to evaluate enhancement effect in serum antioxidant capacity obtained from an antioxidative nutrient-rich enteral diet (AO diet). We also investigated the ability of the AO diet to attenuate lethality, the production of oxidized products, the production of inflammatory cytokines, and liver injury using lipopolysaccharide (LPS)-injected mice. LPS mice were used as a model to represent critically ill patients that have experienced a septicemia. METHODS: The AO diet contained polyphenol and enhanced vitamin C, vitamin E, and trace elements. Total antioxidant activities of the control enteral diet (Control diet) and the AO diet were measured by 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2' azinobis-(3-ethylbenzthiazoline sulphonic acid; ABTS) radical-scavenging activities. Male BALB/c mice were fed either of these diets for 7 days and were injected with 5 mg/kg LPS. The survival of mice was monitored from day 0 to day 8. To evaluate oxidative stress, inflammation, and liver injury, blood and liver samples were collected, and tumor necrosis factor-alpha (TNF-alpha), interleukin 6, thiobarbituric acid-reactive substances (TBARS), protein carbonyl contents, aspartate aminotransferase, alanine aminotransferase, and radical-scavenging activities were measured. RESULTS: The survival rate of mice receiving the AO diet or the Control diet was 73.9% and 33.3%. In the AO diet group, levels of serum TNF-alpha, serum protein carbonyl contents, plasma, and liver TBARS were significantly lower than in the Control diet group. DPPH and ABTS radical scavenging activities of the AO diet itself were significantly higher than that of the Control diet, and serum activities in the AO diet group were also higher. CONCLUSIONS: The antioxidative nutrient supplementation of an enteral diet may be useful and offer relief from septic symptoms. PMID- 17463143 TI - Diethylhexylphthalate extracted by typical newborn lipid emulsions from polyvinylchloride infusion systems causes significant changes in histology of rabbit liver. AB - BACKGROUND: Looking for a candidate substance inducing hepatobiliary dysfunction under parenteral nutrition (PN) in newborns, we recently discovered that newborn infusions extract large amounts of the plasticizer diethylhexylphthalate (DEHP) from commonly used polyvinylchloride (PVC) infusion lines. This plasticizer is well known to be genotoxic and teratogenic in animals and to cause changes in various organs and enzyme systems even in humans. The aim of this study was to examine the effect of DEHP, extracted in the same way and in the same amount as in newborns, on livers of young rabbits. METHODS: Prepubertal rabbits received lipid emulsion through central IV lines continuously for 3 weeks either via PVC or polyethylene (PE) infusion systems. Livers were examined after 1 and 3 weeks by light and electron microscopy. RESULTS: By light microscopy, hydropic degeneration, single-cell necrosis, fibrosis, and bile duct proliferation were observed more in the PVC group. Electron microscopy revealed multiple nuclear changes, clusters and atypical forms of peroxisomes, proliferation of smooth endoplasmic reticulum, increased deposition of lipofuscin, and a mild perisinusoidal fibrosis only in the PVC group. These changes, which are generally regarded as reaction upon a toxic stimulus, could be exclusively attributed to DEHP. CONCLUSIONS: This investigation proved that DEHP produces toxin-like changes in livers of young rabbits in the same dose, duration, and method of administration as in newborn infants. For this reason, it is likely that DEHP is the substance that causes hepatobiliary dysfunction in newborns under PN. Possible modes of action of DEHP are proposed. PMID- 17463144 TI - Lactulose feeding lowers cecal densities of clostridia in piglets. AB - BACKGROUND: In order to understand the consequences of persistent enteral feeding in patients with carbohydrate malabsorption, we fed piglets lactulose in sufficient dosage to produce osmotic diarrhea or inulin, using a conventional dose, to determine if this prebiotic can modulate the effects of lactulose. Feeding lactulose increases cecal luminal synthesis of butyrate, with inulin having an intermediate effect. Because clostridia may be a major source of colonic butyrate production, we hypothesized that feeding piglets lactulose or inulin would increase cecal densities of clostridia. METHODS: Piglets were assigned to 3 formula study groups for 6 days: (1) control, fed only sow milk replacer (n = 12); (2) inulin, inulin supplement (3 g/L; n = 11); and (3) lactulose, lactulose supplement (66.7 g/L; n = 6). Cecal fluid for bacteriological studies was sampled intraoperatively. RESULTS: The wet/dry ratio of the cecal contents (mean +/- SEM) was 8.2 +/- 0.5, 6.2 +/- 0.5, and 18.8 +/- 5.5, respectively, in the control, inulin, and lactulose groups (p = .049, Kruskal-Wallis). There were no differences among the diet groups for cecal densities (10(6) colony-forming units [CFU]/g dry wt cecal contents) of total anaerobes, total aerobes, bifidobacteria, or lactobacilli. Densities of clostridia were markedly reduced in the lactulose group (1.14 +/- 0.41) vs the control (18.39 +/- 4.44; p = .001) or inulin groups (8.87 +/- 2.20; p = .04). CONCLUSIONS: In piglets, feeding lactulose at a dose known to cause diarrhea reduces cecal densities of clostridia. PMID- 17463145 TI - Glutathione decreased parenteral nutrition-induced hepatocyte injury in infant rabbits. AB - BACKGROUND: This study was designed to explore the mechanisms in parenteral nutrition (PN)-associated hepatic dysfunction, and the possible effectiveness of glutathione (GSH) to alleviate this injury. METHODS: Thirty 1-week-old New Zealand rabbits were divided into 3 groups: 10 in the control group (maternal fed); 10 in the PN group (PN for 10 days); and 10 in the GSH + PN group (PN plus glutathione for 10 days). At the end of the study, blood biochemistry analysis and liver histologic examination were performed by light and electronic microscope; malondialdehyde (MDA) content of liver tissues and apoptotic hepatocytes were also measured. RESULTS: Direct bilirubin and bile acid in the PN group were significantly higher than that in the control group and in the GSH + PN group (p < .05, for both). In the PN group, there were some cholestatic or steatotic changes. In the GSH + PN group, histologic changes were reduced compared with the PN group. The electron microscopy appearances were in agreement with the histologic findings. MDA value was higher in the PN group than in the control group and in the GSH + PN group (p < .05, respectively). Terminal deoxynucleotidyl transferase mediated nick end labeling (TUNEL) assays showed that the rate of apoptotic hepatocytes in the PN group was the highest and the control group was the lowest among 3 groups (comparison between groups, p < .01, individually.) CONCLUSIONS: The study showed that PN can induce hepatic dysfunction in infant rabbits. GSH can effectively reduce this injury. The study implies that oxidative stress and apoptosis contribute to PN-associated hepatic dysfunction. PMID- 17463146 TI - Assessment by a multidisciplinary clinical nutrition team before percutaneous endoscopic gastrostomy placement reduces early postprocedure mortality. AB - BACKGROUND: The purpose of this study was to determine whether preassessment by a multidisciplinary nutrition team before percutaneous endoscopic gastrostomy (PEG) placement can reduce postprocedure mortality. This was a prospective single center audit. METHODS: Patients who had been referred to the Gastroenterology Department for consideration of PEG placement between 1995 and 2004 were included. In the index year, 2003-2004, where a formal nutrition team assessment was commenced, 79 patients were enrolled into our study group on a consecutive basis. These patients were subdivided into 3 groups; group A, PEG placed (51 patients); group B, PEG not placed due to severe comorbidity (19 patients); and group C, PEG not placed as deemed unnecessary (9 patients). Comparison was made with previous years where no formal preassessment had occurred. At Staffordshire General Hospital, a comparison of mortality post-PEG placement was made between the index group and previous years. Secondary measures included complication rates and frequency of biochemical monitoring. RESULTS: One week post-PEG mortality fell from 10%-20% in previous years to 0% in the index year (p < .02). This improved survival extended to 3 months postprocedure (p < .016). Three patients (6%) had biochemical evidence of refeeding syndrome postplacement. Biochemical monitoring was inadequate, with only 27/51 (53%) patients being completely monitored. No complications pertaining to the endoscopy were reported. CONCLUSIONS: This study demonstrates that early post-PEG mortality can be reduced by preassessment of patients by a multidisciplinary nutrition team and is evidence supporting the recommendations of the National Confidential Enquiry into Patient Outcome and Death report. PMID- 17463147 TI - Percutaneous endoscopic gastrostomy (PEG) placement in the overweight and obese patient. AB - BACKGROUND: Traditionally, percutaneous endoscopic gastrostomy (PEG) placement in the obese patient has been considered a relative contraindication due to the impedance of gastric transillumination and inability to approximate the abdominal and gastric wall. The aim of this study was to determine the overall success rate, morbidity, and PEG-related mortality in overweight and obese patients. METHODS: Three hundred fifty-five consecutive patients were retrospectively evaluated over a 1-year period at the Virginia Commonwealth University Medical Center for postprocedure-related PEG and PEG/jejunostomy (J) complications. One hundred thirty-four patients were considered overweight (body mass index [BMI] > or = 27 kg/m(2)). Of those, 80 patients were found to be obese (BMI > or = 30 kg/m(2)) with a BMI ranging 30-63 kg/m(2). RESULTS: Gastrostomy placement was successful in 130 of 134 (97%) overweight patients (p < .05). The overall procedure-related mortality was 0%. The rate of significant complications in overweight and obese patients remained 0% when compared with those patients with a normal BMI. Out of 355 patients, 14 failed to receive a PEG; 3 of these were obese and 1 was overweight. These 4 procedures were aborted due to a paucity of anatomical landmarks and failure to transilluminate the abdominal wall. CONCLUSIONS: We believe that PEG placement in the overweight and obese patient can be a technically safe procedure according to our success rate of 100%, with a procedure-related mortality rate of 0%. In those overweight and obese patients who require specialized long-term enteral nutrition support, PEG placement should be considered earlier and more frequently. PMID- 17463148 TI - Predicting energy expenditure in extremely obese women. AB - BACKGROUND: The most common clinical method for resting energy expenditure (REE) assessment is prediction equations. The purpose of this study was to elucidate which prediction equation is most accurate for REE assessment in extremely obese women. METHODS: Fourteen extremely obese women (mean +/- SD body mass index: 49.8 +/- 6.2 kg/m(2); age: 49 +/- 10 years) were measured for height and weight and REE via indirect calorimetry (IC) by a metabolic cart system. Predicted REE was evaluated by several equations, including Harris-Benedict with actual body weight, Harris-Benedict with several adjustments to body weight, Cunningham, Mifflin-St Jeor, Owen, World Health Organization (WHO), and Bernstein equations. Accuracy was determined by mean difference data (IC REE - equation REE; Student's paired t-test), correlation coefficients, and agreement between methods by Bland Altman plots. Accuracy was also evaluated on an individual basis, defined by the percentage of individuals within +/-10% of IC REE. RESULTS: The Mifflin-St Jeor, Harris-Benedict with actual body weight, and the WHO equations were the most accurate in terms of mean predicted REE. The mean predicted REE values by all other equations were different from the IC REE values (p < .1). According to the individual data, the Mifflin-St Jeor was most accurate (14% outside +/-10% IC REE). The Harris-Benedict with actual body weight and WHO equations were less accurate on individual terms, with 29% and 42% of the predicted REE values, respectively, falling outside +/-10% of IC REE. CONCLUSIONS: The Mifflin-St Jeor equation was most accurate method for REE assessment in extremely obese women. PMID- 17463149 TI - Treatment of moderate to severe acute hypocalcemia in critically ill trauma patients. AB - BACKGROUND: Our recent data indicate that 21% of critically ill, adult, multiple trauma patients receiving specialized nutrition support experience hypocalcemia. However, evidence-based methods for the treatment of moderate to severe acute hypocalcemia (ionized calcium concentration [iCa] <1 mmol/L) are lacking. METHODS: The efficacy of an infusion of 4 g of calcium gluconate was evaluated in 20 critically ill, adult, multiple-trauma patients with moderate to severe hypocalcemia (iCa <1 mmol/L). The calcium gluconate was infused at a rate of 1 g/h in a small volume admixture. A serum iCa determination was obtained on the following day. RESULTS: Calcium gluconate infusion significantly increased serum iCa from 0.90 +/- 0.08 mmol/L to 1.16 +/- 0.11 mmol/L (p < .001) on the following day. This dosage regimen was successful for achieving a serum iCa >1 mmol/L for 19 of 20 (95%) hypocalcemic patients and achieved a concentration >1.12 mmol/L in 14 (70%) of the patients. Two patients developed mild hypercalcemia (iCa of 1.34 mmol/L and 1.38 mmol/L) postinfusion. CONCLUSIONS: A short-term infusion of 4 g of intravenous (IV) calcium gluconate for the treatment of moderate to severe hypocalcemia appears to be a promising regimen for critically ill, adult, multiple-trauma patients. PMID- 17463150 TI - Telehealth videoconferencing: improving home parenteral nutrition patient care to rural areas of Ontario, Canada. AB - BACKGROUND: Telehealth videoconferencing is a medium for health care professionals to communicate and care for patients living in remote areas. The aim of this study was to provide a survey to examine management outcome of home parenteral nutrition (HPN) patients when followed by telehealth as an alternative modality of care. METHODS: Twenty-six individuals who were identified to benefit from tele-health were invited to participate in a satisfaction survey. The survey was sent to patients by postal mail. The survey also documented the incidence of line sepsis and other medical HPN complications. A cost analysis was also performed according to technology, human resources, and infrastructure. RESULTS: Eighty-one telehealth videoconference sessions have been held since the inception of telehealth in 2002. Of the current telehealth patients, 13 were eligible for the survey. The satisfaction survey response rate was 11/13 (84.6%). The average line sepsis rate for the 13 patients was 0.89/1000 catheter-days. All patients were generally satisfied with videoconferencing as an alternative method of communication and care for new consultation, patient and family education, and follow-up. Travel time and costs to the patients, their families, and the health care system were significantly less. For example, a patient who resides 611 km from Toronto would cost CDN (Canadian) 724.00 dollars for flight and accommodation to meet with the team at the HPN clinic in Toronto. CONCLUSION: Telehealth incorporated the cost-saving ability for HPN patients to maintain proper medical care, support, and collaboration of specialists inaccessible to their local community. Thus, its strongly positive role in HPN care deserves further consideration for a national application. PMID- 17463151 TI - Noninvasive measurement of transit time in short bowel syndrome. AB - BACKGROUND: Patients with short bowel syndrome (SBS) have limited absorption of nutrients, in part because of the rapidity with which chyme passes through the bowel. We sought to evaluate noninvasive measurements of intestinal transit time to aid in research questions where transit is measured repeatedly. METHODS: Three methods were compared in 8 patients with SBS: the time for blue food color to appear in ostomy effluent or stool, lactulose breath hydrogen testing, and the first radiopaque ring from a capsule in timed stool samples. RESULTS: Median blue dye fasted transit was 96.3 minutes, with breakfast was 117.3 minutes, and with lactulose was 72 minutes. By breath hydrogen testing, transit time was 52.5 minutes but detectible in only 5 of 13 tests. Transit time by radiopaque marker was 967 minutes, but no markers passed in 7 of 14 tests. CONCLUSIONS: Visualization of blue food color in ostomy effluent is a simple, inexpensive approach that can be applied to fasted subjects or after specific meals. Breath hydrogen testing should only be applied in the rare patient with SBS who has an intact ileocecal valve. Radiopaque markers are less useful, most likely because they become trapped due to anatomic changes. PMID- 17463152 TI - Is there evidence that the gut contributes to mucosal immunity in humans? AB - BACKGROUND: Our understanding of the common mucosal immune system derives from animal studies. Antigen-sensitized lymphocytes in the gut-associated lymphoid tissue (GALT) migrate via the blood to mucosal tissues to generate the mucosal associated lymphoid tissue (MALT). In these sites, B cells differentiate into plasma cells and produce antigen-specific secretory IgA, the principal specific immune antiviral and antibacterial defense of moist mucosal surfaces. Responses to oral intake seem necessary to actively maintain this system in health. Experimentally, lack of enteral stimulation with parenteral feeding alters GALT and MALT size and function. These alterations disturb intestinal and extraintestinal mucosal immunity. METHODS: This review is an overview of current and classical studies demonstrating the human mucosal immune system and interactions with nutrition. RESULTS: Human evidence of the mucosal immune system exists, although most data are indirect. Gut stimulation after oral intake induces a generalized immune response in the human MALT through a mucosal-immune network. Examples include neonatal development of GALT influenced by enteral feeding, the presence of antigen-specific IgA and antigen-specific IgA-secreting plasma cells in distant mucosal effector sites such as the breast after gut luminal antigen exposure, and isolation of IgA-producing cells from circulating blood. CONCLUSIONS: It is unlikely that clinical studies will ever completely define the effect of route of feeding in all patient populations. This may be possible, however, if investigators understand, define and characterize nutrition dependent immunologic mechanisms, allowing clinicians to examine clinical responses to nutrition in specific patient populations. This might allow generation of new approaches to protect mucosal immunity. PMID- 17463153 TI - On scientific writing. PMID- 17463155 TI - Oxidative stress, mitochondrial DNA mutation, and apoptosis in aging. AB - A wide spectrum of alterations in mitochondria and mitochondrial DNA (mtDNA) with aging has been observed in animals and humans. These include (i) decline in mitochondrial respiratory function; (ii) increase in mitochondrial production of reactive oxygen species (ROS) and the extent of oxidative damage to DNA, proteins, and lipids; (iii) accumulation of point mutations and large-scale deletions of mtDNA; and (iv) enhanced apoptosis. Recent studies have provided abundant evidence to substantiate the importance of mitochondrial production of ROS in aging. On the other hand, somatic mtDNA mutations can cause premature aging without increasing ROS production. In this review, we focus on the roles that ROS play in the aging-associated decline of mitochondrial respiratory function, accumulation of mtDNA mutations, apoptosis, and alteration of gene expression profiles. Taking these findings together, we suggest that mitochondrial dysfunction, enhanced oxidative stress, subsequent accumulation of mtDNA mutations, altered expression of a few clusters of genes, and apoptosis are important contributors to human aging. PMID- 17463156 TI - Peloruside A, an antimitotic agent, specifically decreases tumor necrosis factor alpha production by lipopolysaccharide-stimulated murine macrophages. AB - Peloruside A (peloruside) is a naturally occurring compound isolated from a New Zealand marine sponge that, like the anticancer drug paclitaxel, stabilizes microtubules and inhibits mitosis. Paclitaxel is known to induce a proinflammatory response in murine macrophages; whereas, peloruside has never been tested for its immunomodulatory effects in these cells. Although the antimitotic effects of the two drugs appear to be similar, we found that peloruside, unlike paclitaxel, does not induce murine macrophages to produce the proinflammatory mediators interleukin-12p40 (IL-12p40), tumor necrosis factor alpha (TNF-alpha), and nitric oxide. The 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide (MTT) reduction assay confirmed that the absence of cytokine production was not caused by cytotoxicity in these nondividing cells. Additionally, there was no effect on unstimulated splenocytes; whereas, both compounds inhibited proliferation after concanalavin A (Con A) stimulation. Finally, there was a significant decrease in TNF-alpha and nitric oxide but not IL-12p40 when macrophages were cultured with lipopolysaccharide (LPS) and either paclitaxel or peloruside. These results suggest that peloruside may prove to be an effective anti-inflammatory treatment, since it does not induce the production of proinflammatory mediators yet can downregulate TNF-alpha and nitric oxide production by LPS-stimulated macrophages, as well as inhibit lymphocyte proliferation. PMID- 17463157 TI - TNF-alpha induces hepatic steatosis in mice by enhancing gene expression of sterol regulatory element binding protein-1c (SREBP-1c). AB - We investigated the effect of tumor necrosis factor-alpha (TNF-alpha), a member of the proinflammatory cytokine family, on steatosis of the mouse liver by analyzing morphological changes and hepatic triglyceride content in response to TNF-alpha. We also examined expression of the sterol regulatory element binding protein-1c gene. Intraperitoneal injection of TNF-alpha acutely and dramatically accelerated the accumulation of fat in the liver, as evidenced by histological analysis and hepatic triglyceride content. This treatment increased liver weight, increased serum levels of free fatty acids, and increased fatty acid synthase and sterol regulatory element binding protein-1c mRNA expression. Furthermore, intraperitoneal injection of lipopolysaccaride (LPS) to induce TNF-alpha expression also accelerated hepatic fat accumulation. Pretreatment with anti-TNF alpha antibody attenuated the development of LPS-induced fatty change in the liver. Antibody pretreatment not only decreased sterol regulatory element binding protein-1c expression in LPS-treated mice but also attenuated the expression of suppressors of cytokine signaling-3 mRNA. This study suggests that TNF-alpha, acting downstream of LPS, increases intrahepatic fat deposition by affecting hepatic lipogenetic metabolism involving sterol regulatory element binding protein-1c. PMID- 17463158 TI - Preservation of hepatocyte nuclear factor-4alpha is associated with zinc protection against TNF-alpha hepatotoxicity in mice. AB - Hepatocyte nuclear factor-4alpha (HNF-4alpha), a zinc finger protein, is the most abundant transcription factor in the liver. HNF-4alpha regulates a large number of genes involved in most aspects of hepatocyte functions. The present study was undertaken to determine the role of HNF-4alpha in zinc protection against tumor necrosis factor-alpha (TNF-alpha) hepatotoxicity. Mice were treated with murine TNF-alpha via intravenous injection at 20 mug/kg body wt 30 mins after d galactosamine (d-Gal) sensitization (800 mg/kg body wt). Two doses of zinc sulfate (5 mg elemental zinc/kg body wt) were administered at 36 and 12 hrs before TNF-alpha treatment via subcutaneous injection. TNF-alpha treatment after sensitization induced liver injury as detected by plasma alanine aminotransferase activity and apoptotic cell death in the liver. Zinc pretreatment attenuated TNF alpha-induced liver injury. Furthermore, TNF-alpha-induced activations of caspase 3 and caspase 8 in the liver were significantly inhibited by zinc pretreatment. The mRNA and protein levels of HNF-4alpha in the liver were remarkably decreased by TNF-alpha treatment, which was suppressed by zinc. To determine if HNF-4alpha depletion is involved in d-Gal sensitization to TNF-alpha toxicity, mice were administered either d-Gal or TNF-alpha. Immunohistochemistry demonstrated that HNF-4alpha depletion in the liver is associated with d-Gal sensitization but not TNF-alpha treatment. To define the link between HNF-4alpha depletion and TNF alpha-induced cell death, the effect of silencing the HNF-4alpha gene by siRNA transfection on TNF-alpha cytotoxicity was determined in HepG2 cells. A lactate dehydrogenase cytotoxicity assay showed that neither TNF-alpha nor HNF-4alpha siRNA transfection had a toxic effect, but TNF-alpha treatment after HNF-4alpha siRNA transfection caused HepG2 cell death. These results suggest that zinc protects against TNF-alpha hepatotoxicity, at least partially, through preservation of the zinc finger protein HNF-4alpha. PMID- 17463159 TI - Effect of glycosaminoglycans on matrix metalloproteinases in type II collagen induced experimental arthritis. AB - Matrix metalloproteinases (MMPs) are a family of neutral proteinases that are involved in tissue remodeling by mediating degradation of extracellular matrix components in both physiology and pathology. As MMPs appear to play a key role in the degradation of cartilage matrix in the progression of arthritic disease, MMPs are considered as potential therapeutic targets. The effect of chondroitin sulfate A (CSA) on MMPs in type II collagen-induced experimental arthritis was studied. The anti-arthritic effect of CSA was evidenced by a decrease in marker activities like lysosomal beta-hexosaminidase and beta-glucuronidase. Arthritic animals showed significantly higher activity of MMP2 and MMP9 and increased levels of other MMPs, including MMP3 and MT-1 MMP in cartilage and serum. Treatment with CSA significantly decreased the activity of MMPs, particularly MMP9 in serum and synovial effusate and cartilage. The effect of CSA was further studied by fragmenting CSA into low-molecular-weight oligosaccharides. The oligosaccharide-treated animals showed considerably lower MMP activity (particularly MMP9) compared with arthritic controls. The CSA (and the oligosaccharides derived from it) not only reduced the activity of MMPs but also decreased the protein level expression of MMPs, indicating that the production of MMPs is affected. These results indicate that the antiarthritic effect of CSA involves down-regulation of MMPs, which are critically involved in the progression of arthritic disease. PMID- 17463160 TI - Correlation between mRNA expression level of the mutant COL4A5 gene and phenotypes of XLAS females. AB - Alport syndrome (AS) is a progressive hereditary glomerulonephritis presented with hematuria, progressive renal failure, sensorineural deafness, and ocular lesions. Females with X-linked Alport syndrome (XLAS) have variable phenotypes, from asymptomatic hematuria to renal failure. In order to understand the possible mechanism of different phenotypes in female XLAS, we analyzed mRNA expression level of the mutant COL4A5 gene in fibroblasts, the X-inactivation pattern in peripheral blood DNA, and the phenotype variability of XLAS females. Total RNA was isolated from cultured skin fibroblasts in five females with XLAS and confirmed deletion mutations of COL4A5 mRNA. Reverse transcription-polymerase chain reaction (PCR) was performed to amplify the fragment, including the mutation sequences of the COL4A5 gene. The PCR products were electrophoresed with 8% polyacrylamide gel. Messenger RNA expression level of the mutant COL4A5 gene was analyzed with the optical density of PCR product revealed under polyacrylamide gel. The X-inactivation analysis was performed using HpaII predigestion of peripheral blood DNA followed by PCR of the highly polymorphic CAG repeat of the androgen receptor (AR) gene. All patients in the study had persistent microscopic hematuria. Two of them had gross hematuria. Three cases had persistent and severe proteinuria of 2+~3+, and the others had discontinuous and milder proteinuria of - ~+. The patients whose mRNA expression level of the mutant COL4A5 gene was higher had persistent and more severe proteinuria (r = 0.975, P = 0.005). None of them had skewed X inactivation. Our preliminary results demonstrate that the quantity of mRNA expression level of the mutant COL4A5 gene was correlated with the phenotypic severity of females with XLAS, and this could not be explained by X-inactivation pattern in peripheral blood leukocytes. PMID- 17463161 TI - Celecoxib decreases fatty acid synthase expression via down-regulation of c-Jun N terminal kinase-1. AB - Nonsteroidal anti-inflammatory drugs (NSAIDs) are inhibitors of cyclooxygenase (COX). Our previous observations that celecoxib, a COX-2-specific inhibitor, not only inhibits rat mammary carcinogenesis, but also decreases fat deposition in rats fed a high-fat diet, prompted us to determine whether celecoxib affects lipid metabolism. At 57 days of age, two groups of 10 female Sprague Dawley rats were pair-fed a high-fat diet with or without 1500 ppm celecoxib for 15 weeks. Compared with controls, celecoxib-treated rats had 44.4% less hepatic triglycerides and 22.6% less intra-abdominal adipose tissue mass. In the liver and adipose tissue of several genes involved in fat metabolism and mobilization that we measured, only fatty acid synthase (FAS) was significantly down-regulated by celecoxib treatment. There were no differences in the level of prostaglandin E(2) in these tissues, indicating that celecoxib decreases fat accumulation by down-regulating FAS through a COX-2-independent mechanism. Among the potential molecular targets by which celecoxib may regulate FAS expression, only c-Jun N terminal kinase-1 (JNK1) was significantly down-regulated. Furthermore, a known inhibitor of JNK suppressed FAS expression in rat hepatocytes. Our observations suggest that celecoxib suppresses FAS expression and decreases fat accumulation by down-regulating JNK1. PMID- 17463162 TI - Vanadate elevates fetal hemoglobin in human erythroid precursors by inhibiting cell maturation. AB - Increased fetal hemoglobin (HbF) in erythroid precursors of patients with beta hemoglobinopathies (sickle cell anemia and beta-thalassemia), in which adult hemoglobin synthesis is defective, ameliorates the clinical symptoms of the underlying diseases. The production of erythroid precursors depends on the action of erythropoietin (EPO), which prevents their apoptosis and stimulates their proliferation. EPO binds to its surface receptor, induces its homodimerization, and initiates a cascade of phosphorylation and dephosphorylation of a series of proteins by kinases and phosphatases, respectively. Vanadate inhibits various phosphatases, including those that are involved in the EPO pathway, thereby intensifying the signal. In this study, we investigated the effect of vanadate on the proliferation and maturation of human erythroid precursors in culture. When vanadate was added to cells derived from normal donors, cell maturation was delayed, as indicated by cell morphology, cell growth kinetics, the rate of appearance of hemoglobin-containing cells, and the expression of surface antigens (CD117, CD71, and glycophorin A). Analysis by high-performance liquid chromatography and flow cytometry of the hemoglobin profile of vanadate-treated normal cells revealed a higher proportion of HbF than was found in untreated cells. When vanadate was added to cells derived from patients with beta thalassemia, a significant increase in HbF was observed. The results suggest that intensification of the EPO signal by vanadate results in maturation arrest and increased HbF production. Thus, inhibitors that are more specific and less toxic than vanadate may present a novel option for elevating HbF in patients with beta hemoglobinopathies, as well as for intensifying the EPO response in other forms of anemia. PMID- 17463163 TI - Novel localization of NMDA receptors within neuroendocrine gonadotropin-releasing hormone terminals. AB - About 1000 hypothalamic neurons synthesize and release gonadotropin-releasing hormone (GnRH), the master molecule of reproduction in all mammals. At the level of the median eminence at the base of the brain, where GnRH and other hypothalamic releasing hormones are secreted into the capillary system leading to the anterior pituitary gland, there is non-synaptic regulation of neurohormone release by a number of central neurotransmitters. For example, glutamate, the major excitatory amino acid in the brain, directly regulates GnRH release from nerve terminals via NMDA receptors (NMDARs). Moreover, the effects of glutamate action on GnRH secretion are potentiated by estrogens, and this relates to the physiologic control of ovulation by the hypothalamus. We sought to determine the ultrastructural relationship between GnRH neuroterminals and NMDARs, and this regulation by estradiol. Using immunofluorescent confocal microscopy, postembedding immunogold electron microscopy, fractionation, and Western blotting, we demonstrated: (i) GnRH is localized in large dense-core vesicles of neurosecretory profiles/terminals, (ii) the NMDAR1 subunit is found primarily on large dense-core vesicles of neurosecretory profiles/terminals, (iii) there is extensive colocalization of GnRH and NMDAR1 on the same vesicles, and (iv) estradiol modestly but significantly alters the distribution of NMDAR1 in GnRH neuroterminals by increasing expression of NMDAR1 on large dense-core vesicles. Western blots of fractionated median eminence support the presence of NMDAR1 in subcellular fractions containing large dense-core vesicles. These data are the first to show the presence of the NMDAR on neuroendocrine secretory vesicles, its co-expression with GnRH, and its regulation by estradiol. The results provide a novel anatomical site for the NMDAR and may represent a new mechanism for the regulation of GnRH release. PMID- 17463164 TI - The soy effect in the disease models of nonbacterial prostatitis and obstructive voiding. AB - The goal of this study was to improve the understanding of the potential significance of dietary soy for human health by investigating its effects in the animal models of nonbacterial prostatitis and urethral obstruction. Nonbacterial prostatitis was induced in adult Noble rats with the combined treatment of testosterone and 17beta-estradiol. The inflammatory foci categorized into three forms were counted and correlated with expression of an estrogen-responsive gene, progesterone receptor (PR), in the dorsolateral lobes of the rats on soy (+) and soy (-) diets. Development of obstructive voiding after neonatal estrogenization of Noble rats (NeoDES rats) was followed with urodynamic measurements in rats on soy (+) and soy (-) diets. The amounts of genistein and daidzein, two major soy derived isoflavones, were measured in the urine of Noble rats by the high performance liquid chromatography-photodiodearray method. Dietary soy decreased the total number of inflammatory foci while no demonstrable effects were seen on the cellular composition of the infiltrates. Soy did not increase the weights of the pituitary gland, testes, or sex accessory glands, but it did increase the number of PR-positive epithelial cells in the dorsolateral prostate. It also decreased the bladder pressures in NeoDES rats but did not increase the flow rates. The soy effects may be mediated by the strong estrogen influence involved in the animal models. Dietary soy had anti-inflammatory effects in the prostate but only marginal effects on the development of obstructive voiding in Noble rats. The anti-inflammatory effects of soy may contribute to the lower prevalence of prostatitis-like symptoms and the historically lower risk of benign prostatic hyperplasia in Japan; however, no evidence was found that regular consumption of soy influences the age-related development of lower urinary tract symptoms or decline of flow rate. PMID- 17463165 TI - Zinc inhibits doxorubicin-activated calcineurin signal transduction pathway in H9c2 embryonic rat cardiac cells. AB - Elevation of the zinc-binding protein metallothionein (MT) in the heart inhibits doxorubicin (DOX)-induced myocardial apoptosis and heart hypertrophy. Zinc release from MT in response to oxidative stress has been suggested as a mechanism of action of MT protection from DOX toxicity, and calcineurin is involved in the signaling pathways leading to myocardial apoptosis and heart hypertrophy. The present study was undertaken to determine if zinc can modulate the DOX-activated calcineurin signaling pathway. H9c2 cells were treated with 1 muM DOX, and zinc release was monitored by a zinc ion-specific fluorophore, zinquin ethyl ester. Additionally, DOX-activated calcineurin signaling was detected by a calcineurin dependent nuclear factor of activated T-cell reporter. DOX treatment induced an increase in intracellular labile zinc and activated calcineurin signaling. Pretreatment of H9c2 cells with a zinc-specific, membrane-permeable chelating agent, N,N,N',N'-tetrakis(2-pyridylmethyl)ethylenediamine (TPEN), inhibited the increase in intracellular labile zinc and increased the DOX-activated calcineurin signaling. Pretreatment of H9c2 cells with exogenously added zinc attenuated the DOX-activated calcineurin signaling in a dose-dependent manner. However, neither TPEN nor addition of exogenous zinc affected DOX-induced cellular hypertrophy or DOX-induced decrease in cell viability. Additionally, inhibition of DOX-induced calcineurin signaling with the calcineurin inhibitors cyclosporine A or tacrolimus (FK506) failed to restrict the DOX-induced decrease in cell viability. These results indicate that zinc suppresses DOX-induced calcineurin signaling in H9c2 cells; however, calcineurin signaling is not involved in the DOX-induced decrease in cell viability in H9c2 cells. (It had been shown previously that calcineurin is also not necessary for DOX-induced H9c2 cell hypertrophy.). PMID- 17463166 TI - Role of NO and COX pathways in mediation of adenosine A1 receptor-induced renal vasoconstriction. AB - The mechanism of adenosine A1 receptor-induced intrarenal vasoconstriction is unclear; it depends on sodium intake and may be mediated by changing the intrarenal activity of the nitric oxide (NO) and/or cyclooxygenase (COX) pathway of arachidonic acid metabolism. The effects of 2-chloro-N(6)-cyclopentyl adenosine (CCPA), a selective A1 receptor agonist, on renal hemodynamics were examined in anesthetized rats maintained on high sodium (HS) or low sodium (LS) diet. Total renal (i.e., cortical) blood flow (RBF) as well as superficial cortical (CBF), outer medullary (OMBF), and inner medullary (IMBF) flows were determined by laser-Doppler. In HS rats, suprarenal aortic infusions of 8-40 nmol/kg/hr CCPA decreased IMBF (15%) and other perfusion indices (22%-27%); in LS rats, IMBF increased 3% (insignificant) and other indices decreased 13%-24%. In LS rats, pretreatment with N-nitro-L-arginine methyl ester prevented the A1 receptor-mediated decrease in RBF and CBF but not OMBF; the response in IMBF was not altered. Pretreatment with indomethacin prevented the decreases in RBF, CBF, and OMBF and did not change the response of IMBF. Thus, within the cortex the vasoconstriction that follows A1 receptor activation results both from inhibition of NO synthesis and from stimulation of vasoconstrictor products of the COX pathway. In the outer medulla, the latter products seem exclusively responsible for CCPA-induced vasoconstriction. The observation that in LS rats IMBF was not affected by stimulation of adenosine A1 receptors suggests that limiting salt intake may help protect medullary perfusion against vasoconstrictor stimuli which have the potential to disturb long-term control of arterial pressure. PMID- 17463167 TI - The novel tetratricopeptide repeat domain 7 mutation, Ttc7fsn-Jic, with deletion of the TPR-2B repeat causes severe flaky skin phenotype. AB - We carried out molecular analyses of the novel flaky skin mutation, Ttc7(fsn-Jic )(a synonym for fsn(Jic)), which we found in a previous study. It was revealed that this mutation involved a genomic in-frame deletion including exons 9 and 10 of the Ttc7 gene, and that the genomic deletion in Ttc7 (fsn-Jic )may disrupt the tetratricopeptide repeat-2B domain of the TTC7 protein. Based on a comparison of three Ttc7 mutations, including Ttc7(fsn-J )(a synonym for fsn) and Ttc7(fsn-hea )(a synonym for hea), it was suggested that either exon 9 or exon 10 or both may play a more important role than the other exons of the Ttc7 gene. Ttc7 gene expression analyses using Northern blotting revealed that Ttc7 mRNA is expressed in 11 tissues, except muscle. In conclusion, we confirmed that the Ttc7 (fsn-Jic )mutation, as well as the Ttc7(fsn-J )and Ttc7 (fsn-hea )mutations, is responsible for abnormal phenotypes observed in various tissues of mice with the flaky skin mutation. PMID- 17463168 TI - Protective role of connexin 32 in steady-state hematopoiesis, regeneration state, and leukemogenesis. AB - The role of gap junctions formed by connexins (Cxs) has been implicated in the homeostatic regulation of multicellular systems. Primitive hematopoietic progenitor cells form a multicellular system, but a previous report states that Cx32 is not expressed in the bone marrow. Thus, a question arises as to why Cx molecules are not detected in the hematopoietic tissue other than in stromal cells. Based on our preliminary study, which suggested a potential impairment of hematopoiesis in Cx32-knockout (KO) mice, the objectives of the present study were to determine whether Cx32 functions in the bone marrow during steady-state hematopoiesis and to examine its possible protective roles during regeneration after chemical abrasions and during leukemogenesis after the administration of a secondary genotoxic chemical, methyl nitrosourea (MNU). As a result, the Cx32 molecule, functioning in the hematopoietic stem cell (HSC) compartment during steady-state hematopoiesis, was observed for the first time; the expressions of Cx32 at the mRNA level, as determined by polymerase chain reaction analysis, and at the protein level, determined using an anti-Cx32 antibody, were observed only in the lin(-)c-kit(+) HSC fraction, using a combination of immunobead-density gradient and immunomagnetic bead separation. Hematopoiesis was impaired in the absence of Cx32, and it was delayed during regeneration after chemical abrasion with 5-fluorouracil at 150 mg/kg body wt in Cx32-KO mice. Cx32-KO mice showed increased leukemogenicity compared with wild-type mice after MNU injection; furthermore, in a competitive assay for leukemogenicity in mice that had been lethally irradiated and repopulated with a mixed population of bone marrow cells from Cx32-KO mice and wild-type mice, the resulting leukemias originated predominantly from Cx32-KO bone marrow cells. In summary, the role of Cx32 in hematopoiesis was not previously recognized, and Cx32 was expressed only in HSCs and their progenitor cells. The results indicate that Cx32 in wild-type mice protects HSCs from chemical abrasion and leukemogenic impacts. PMID- 17463169 TI - Deficient CD4+ CD25+ FOXP3+ T regulatory cells in acquired aplastic anemia. AB - Regulatory T cells are believed to control the development and progression of autoimmunity by suppressing autoreactive T cells. Decreased numbers of CD4(+)CD25(+) FOXP3(+) T cells (Tregs) are associated with impaired immune homeostasis and development of autoimmune diseases. The transcription factors FOXP3 and NFAT1 have key roles in regulatory T-cell development and function. We show that Tregs are decreased at presentation in almost all patients with aplastic anemia; FOXP3 protein and mRNA levels also are significantly lower in patients with aplastic anemia and NFAT1 protein levels are decreased or absent. Transfection of FOXP3-deficient CD4(+)CD25(+) T cells from patients with a plasmid encoding wild-type NFAT1 resulted in increased FOXP3 expression in these cells. By NFAT1 knockdown in CD4(+)CD25(+) T cells, FOXP3 expression was decreased when NFAT1 expression was decreased. Our findings indicate that decreased NFAT1 could explain low FOXP3 expression and diminished Treg frequency in aplastic anemia. Treg defects are now implicated in autoimmune marrow failure. PMID- 17463170 TI - Role of CD28 in fatal autoimmune disorder in scurfy mice. AB - Scurfy mice develop CD4 T-cell-mediated lymphoproliferative disease leading to death within 4 weeks of age. The scurfy mutation causes loss of function of the foxp3 gene (foxp3(sf)), which is essential for development and maintenance of naturally occurring regulatory CD4 T cells (nTregs). In humans, mutations of the foxp3 gene cause immune dysregulation, polyendocrinopathy, enteropathy, and X linked syndrome (IPEX). In most patients with IPEX and also in scurfy mice, T cells show hyperreactivity and levels of Th1- and Th2-associated cytokines are substantially elevated. We report that removal of CD28 expression rescued scurfy mice from early death. Longer-term surviving CD28-deficient scurfy mice still had lymphoproliferative disorder, but their CD4 T cells showed decreased interferon gamma and no sign of interleukin-4 or interleukin-10 hyperproduction. Furthermore, injection of CTLA4-Ig to block CD28-B7 interactions substantially improved the survival of scurfy mice by blocking effector T-cell differentiation. These data support the hypothesis that CD28-B7 interactions play a critical role in the etiology of lethal autoimmune disease in scurfy mice by stimulating the differentiation of antigen-activated naive T cells into effector T cells. PMID- 17463171 TI - Inhibition of glycogen synthase kinase-3 activity leads to epigenetic silencing of nuclear factor kappaB target genes and induction of apoptosis in chronic lymphocytic leukemia B cells. AB - Chronic lymphocytic leukemia (CLL) is commonly defined as a disease of failed apoptosis of B cells and remains an incurable disease. The mechanism of resistance to apoptosis in CLL is complex and influenced by numerous factors, including nuclear factor kappaB (NFkappaB)-mediated expression of antiapoptotic molecules. Recent evidence indicates that glycogen synthase kinase-3beta (GSK 3beta) positively regulates NFkappaB-mediated gene transcription and cell survival. Using malignant B cells collected from patients with CLL, we find that both GSK-3beta and NFkappaB accumulate in the nucleus of CLL B cells, and pharmacologic inhibition of GSK-3 results in decreased expression of two NFkappaB target genes Bcl-2 and XIAP and a subsequent increase in CLL B-cell apoptosis ex vivo. Furthermore, we observed that inhibition of GSK-3 leads to a decrease in NFkappaB-mediated gene transcription but does not affect the nuclear accumulation of NFkappaB in CLL B cells. Last, using chromatin immunoprecipitation, we show that GSK-3 inhibition abrogates NFkappaB binding to its target gene promoters (XIAP, Bcl-2), in part through epigenetic modification of histones. Our results establish that inhibition of GSK-3 abrogates NFkappaB binding to its target gene promoters through an epigenetic mechanism, enhances apoptosis in CLL B cells ex vivo and identifies GSK-3 as a potential therapeutic target in the treatment of CLL. PMID- 17463172 TI - Comparison of bromcresol green and agarose protein electrophoresis for quantitation of serum albumin in multiple myeloma. AB - BACKGROUND: The International Staging System for multiple myeloma has increased the importance of accurate measurement of serum albumin. Two common albumin assays, bromcresol green (BCG) and agarose gel protein electrophoresis (PEL), frequently yield discordant results, creating confusion regarding which assay is superior for use in myeloma. METHODS: We measured albumin by BCG on a Roche Modular system, by PEL with a Helena SPIFE SPE Vis agarose gel, and by immunonephelometry performed on a Dade Behring BNII nephelometer. BCG and PEL were used to measure albumin in 5777 patient samples, and all 3 methods were used in an additional 252 samples. The clinical impact was assessed on 698 myeloma patient samples. RESULTS: For sera with zero/low monoclonal immunoglobulin protein (M)-spike (0 to <15 g/L), results for both BCG and PEL correlated well to nephelometry, although median PEL results were 8 g/L lower than corresponding BCG measurements. Correlation between PEL and nephelometry or BCG diminished with increasing M-spike, with PEL eventually overestimating albumin compared with both other assays. IgG and IgA M-spikes showed significantly different effects on albumin discordance. For 35% of myeloma patients, discrepancy between BCG and PEL had a potentially clinically significant effect on staging, but no difference in group survival was found. CONCLUSIONS: Both BCG and PEL correlate well to nephelometry in sera with zero/low M-spikes. In the presence of larger M-spikes, PEL correlates poorly to nephelometry or BCG, whereas BCG compares well with nephelometry regardless of M-spike. Thus, albumin measurement can be performed reliably in myeloma patient sera by use of inexpensive, automated BCG assays. PMID- 17463173 TI - Aptamers evolved from cultured cancer cells reveal molecular differences of cancer cells in patient samples. AB - BACKGROUND: Molecular-level differentiation of neoplastic cells is essential for accurate and early diagnosis, but effective molecular probes for molecular analysis and profiling of neoplastic cells are not yet available. We recently developed a cell-based SELEX (systematic evolution of ligands by exponential enrichment) strategy to generate aptamers (designer DNA/RNA probes) as molecular probes to recognize neoplastic cells. METHODS: We tested 6 cell-SELEX-generated aptamers with equilibrium dissociation constants in the nanomolar to subnanomolar range: sgd5, selected from Toledo cells, a human diffuse large-cell lymphoma cell line (B-cell), and sgc8, sgc3, sgc4, sgd2, and sgd3 from CCRF-CEM cells, a human precursor T cell acute lymphoblastic leukemia (T-ALL) cell line. Aptamers were labeled with fluorescein isothiocyanate fluorophores and then used to recognize, by flow cytometric analysis, neoplastic cells in cultured hematopoietic cell lines and clinical samples. RESULTS: Aptamer sgd5 recognized only its target cells. Aptamers sgc3, sgd2, sgd3, sgc4, and sgc8, selected from a T-cell leukemia cell line, identified all of the cultured T-cell leukemia cell lines with relatively high fluorescence intensity. Aptamers sgc8, sgc3, and sgd3 showed good selectivity toward T-ALL cells and almost no binding to normal hematopoietic cells or lymphoma and myeloma cells. Selected aptamers also detected targets on the cell membranes of neoplastic cells in patient samples. CONCLUSIONS: Aptamers selected against cultured neoplastic cells can effectively be used as molecular probes for recognition of neoplastic cells in patient samples. Cell-based aptamer selection can be used to generate aptamer probes to obtain molecular signatures of neoplastic cells in patient samples. PMID- 17463174 TI - Real-time PCR determination of IMPDH1 and IMPDH2 expression in blood cells. AB - BACKGROUND: Inosine monophosphate dehydrogenase (IMPDH) catalyzes the rate limiting step in de novo guanine nucleotide synthesis and is implicated in cell cycle control. Inhibition of this enzyme is associated with immunosuppressive, antiviral, and antitumor activity. IMPDH basal activity increases after initiation of immunosuppressive therapy. METHODS: A real-time reverse transcription PCR assay was developed and validated for mRNA quantification of the 2 human IMPDH isoforms. Target gene expressions were normalized to the geometric mean of 3 housekeeping genes. Assay utility was tested by analyzing patient samples and cultured cells exposed to immunosuppressive drugs such as the IMPDH inhibitor mycophenolic acid. RESULTS: The assay was linear over 6 logs of cDNA input and demonstrated specific quantification of IMPDH1 and IMPDH2 expression in cultured cells and patient samples. Limits of detection and quantification were 10 and 10(3) copies of cDNA per reaction, respectively. Within-run and total between-day CVs were <15% for normalized expression. Changes in IMPDH1 and 2 expression were observed in patient samples after initiation of an immunosuppressive regimen that included calcineurin inhibitors, mycophenolate mofetil, and steroids. CONCLUSIONS: This assay can be used to study the regulation of IMPDH expression and the involvement of the enzymes in immunological and malignant proliferative conditions. This may contribute to the processes of drug development and to the establishment of monitoring strategies for treatment effect and disease activity. PMID- 17463175 TI - A novel, high-throughput workflow for discovery and identification of serum carrier protein-bound peptide biomarker candidates in ovarian cancer samples. AB - BACKGROUND: Most cases of ovarian cancer are detected at later stages when the 5 year survival is approximately 15%, but 5-year survival approaches 90% when the cancer is detected early (stage I). To use mass spectrometry (MS) of serum proteins for early detection, a seamless workflow is needed that provides an opportunity for rapid profiling along with direct identification of the underpinning ions. METHODS: We used carrier protein-bound affinity enrichment of serum samples directly coupled with MALDI orthagonal TOF MS profiling to rapidly search for potential ion signatures that contained discriminatory power. These ions were subsequently directly subjected to tandem MS for sequence identification. RESULTS: We discovered several biomarker panels that enabled differentiation of stage I ovarian cancer from unaffected (age-matched) patients with no evidence of ovarian cancer, with positive results in >93% of samples from patients with disease-negative results and in 97% of disease-free controls. The carrier protein-based approach identified additional protein fragments, many from low-abundance proteins or proteins not previously seen in serum. CONCLUSIONS: This workflow system using a highly reproducible, high-resolution MALDI-TOF platform enables rapid enrichment and profiling of large numbers of clinical samples for discovery of ion signatures and integration of direct sequencing and identification of the ions without need for additional offline, time-consuming purification strategies. PMID- 17463176 TI - Natural calcium isotopic composition of urine as a marker of bone mineral balance. AB - BACKGROUND: We investigated whether changes in the natural isotopic composition of calcium in human urine track changes in net bone mineral balance, as predicted by a model of calcium isotopic behavior in vertebrates. If so, isotopic analysis of natural urine or blood calcium could be used to monitor short-term changes in bone mineral balance that cannot be detected with other techniques. METHODS: Calcium isotopic compositions are expressed as delta(44)Ca, or the difference in parts per thousand between the (44)Ca/(40)Ca of a sample and the (44)Ca/(40)Ca of a standard reference material. delta(44)Ca was measured in urine samples from 10 persons who participated in a study of the effectiveness of countermeasures to bone loss in spaceflight, in which 17 weeks of bed rest was used to induce bone loss. Study participants were assigned to 1 of 3 treatment groups: controls received no treatment, one treatment group received alendronate, and another group performed resistive exercise. Measurements were made on urine samples collected before, at 2 or 3 points during, and after bed rest. RESULTS: Urine delta(44)Ca values during bed rest were lower in controls than in individuals treated with alendronate (P <0.05, ANOVA) or exercise (P <0.05), and lower than the control group baseline (P <0.05, t-test). Results were consistent with the model and with biochemical and bone mineral density data. CONCLUSION: Results confirm the predicted relationship between bone mineral balance and calcium isotopes, suggesting that calcium isotopic analysis of urine might be refined into a clinical and research tool. PMID- 17463177 TI - Analytical validation of the Oncotype DX genomic diagnostic test for recurrence prognosis and therapeutic response prediction in node-negative, estrogen receptor positive breast cancer. AB - BACKGROUND: Oncotype DX is a clinically validated, high-complexity, multianalyte reverse transcription-PCR genomic test that predicts the likelihood of breast cancer recurrence in early-stage, node-negative, estrogen receptor-positive breast cancer. The Recurrence Score (RS) provides a more accurate, reproducible measure of breast cancer aggressiveness and therapeutic responsiveness than standard measures. Individualized patient management requires strict performance criteria for clinical laboratory tests. We therefore investigated the analytical performance of the assay. METHODS: Assays used a pooled RNA sample from fixed paraffin-embedded tissues to evaluate the analytical performance of a 21-gene panel with respect to amplification efficiency, precision, linearity, and dynamic range, as well as limits of detection and quantification. Performance variables were estimated from assays carried out with sample dilutions. In addition, individual patient samples were used to test the optimized assay for reproducibility and sources of imprecision. RESULTS: Assay results defined acceptable operational performance ranges, including an estimated maximum deviation from linearity of <1 cycle threshold (C(T)) units over a > or =2000 fold range of RNA concentrations, with a mean quantification bias of 0.3% and CVs of 3.2%-5.7%. An analysis of study design showed that assay imprecision contributed by instrument, operator, reagent, and day-to-day baseline variation was low, with SDs of <0.5 C(T). CONCLUSION: The analytical and operational performance specifications defined for the Oncotype DX assay allow the reporting of quantitative RS values for individual patients with an SD within 2 RS units on a 100-unit scale. PMID- 17463178 TI - Platelet resistance to the antiaggregatory cyclic nucleotides in central obesity involves reduced phosphorylation of vasodilator-stimulated phosphoprotein. AB - BACKGROUND: Impairment of platelet response to antiaggregatory agents is seen in individuals with central obesity and may play a role in the increased cardiovascular risk associated with obesity. In this study we evaluated whether this impairment involves the antiaggregatory pathways regulated by cAMP and cGMP. METHODS: We obtained platelet-rich plasma from 12 obese individuals and 12 controls. We investigated the effects of the cyclic nucleotide analogs 8-pCPT cAMP (10-500 micromol/L) and 8-pCPT-cGMP (10-500 micromol/L) on ADP-induced platelet aggregation as assessed by decreased light scattering. We assessed the activation of cAMP- and cGMP-dependent protein kinases by measuring phosphorylation of the vasodilator-stimulated phosphoprotein (VASP) at Ser157 and Ser239. RESULTS: The antiaggregatory effect of both cyclic nucleotide analogs was impaired in obese individuals compared to controls, with mean (SE) half-maximal inhibitory concentrations (IC(50)) (after 20-min incubation) of 123 (33) micromol/L vs 5 (1) micromol/L, respectively, for 8-pCPT-cAMP (P <0.01) and of 172 (43) micromol/L vs 17 (8) micromol/L, respectively, for 8-pCPT-cGMP (P <0.01). The Homeostasis Model Assessment Index of Insulin Resistance was independently correlated with cyclic nucleotide analog IC(50). In obese individuals, VASP phosphorylation at Ser157 and Ser239 in response to cyclic nucleotides was significantly lower than in controls. CONCLUSIONS: In central obesity the reduced ability of cyclic nucleotides to inhibit platelet aggregation is associated with reduced activation of their specific kinases. Because cyclic nucleotides help regulate platelet antiaggregation, alteration of this ability is consistent with platelet hyperactivity in obesity. PMID- 17463179 TI - The homeodomain transcription factors Cdx1 and Cdx2 induce E-cadherin adhesion activity by reducing beta- and p120-catenin tyrosine phosphorylation. AB - The homeodomain transcription factors Cdx1 and Cdx2 are regulators of intestine specific gene expression. They also regulate intestinal cell differentiation and proliferation; however, these effects are poorly understood. Previously, we have shown that expression of Cdx1 or Cdx2 in human Colo 205 cells induces a mature colonocyte morphology characterized by the induction of a polarized, columnar shape with apical microvilli and strong cell-cell adhesion. To elucidate the mechanism underlying this phenomenon, we investigated the adherens junction complex. Cdx1 or Cdx2 expression reduced Colo 205 cell migration and invasion in vitro, suggesting a physiologically significant change in cadherin function. However, Cdx expression did not significantly effect E-cadherin, alpha-, beta-, or gamma-catenin, or p120-catenin protein levels. Additionally, no alteration in their intracellular distribution was observed. Cdx expression did not alter the coprecipitation of beta-catenin with E-cadherin; however, it did reduce p120 catenin-E-cadherin coprecipitation. Tyrosine phosphorylation of beta- and p120 catenin is known to disrupt E-cadherin-mediated cell adhesion and is associated with robust p120-catenin/E-cadherin interactions. We specifically investigated beta- and p120-catenin for tyrosine phosphorylation and found that it was significantly diminished by Cdx1 or Cdx2 expression. We restored beta- and p120 catenin tyrosine phosphorylation in Cdx2-expressing cells by knocking down the expression of protein tyrosine phosphatase 1B and noted a significant decline in cell-cell adhesion. We conclude that Cdx expression in Colo 205 cells induces E cadherin-dependent cell-cell adhesion by reducing beta- and p120-catenin tyrosine phosphorylation. Ascertaining the mechanism for this novel Cdx effect may improve our understanding of the regulation of cell-cell adhesion in the colonic epithelium. PMID- 17463180 TI - Disruption of function and localization of tight junctional structures and Mrp2 in sustained estradiol-17beta-D-glucuronide-induced cholestasis. AB - Estradiol-17beta-D-glucuronide (E(2)17G) induces immediate and profound but transient cholestasis in rats when administered as a single bolus dose. Here, we examined the consequence of sustained E(2)17G cholestasis and assessed the function and localization of the tight junctional proteins zonula occludens-1 (ZO 1) and occludin and of the canalicular transporter multidrug resistance associated protein-2 (Mrp2). An initial dose of E(2)17G (15 mumol/kg iv) followed by five subsequent doses of 7.5 mumol/kg from 60 to 240 min induced a sustained 40-70% decrease in bile flow. Following their biliary retrograde administration, cholera toxin B subunit-FITC or horseradish peroxidase were detected at the sinusoidal domain, indicating opening of the paracellular route; this occurred as early as 15 min after the first dose as well as 15 min after the last dose of E(2)17G, but not following the administration of vehicle in controls. Localization of ZO-1 and occludin was only slightly affected under acute cholestatic conditions but was severely disrupted under sustained cholestasis, with their appearance suggesting a fragmented structure. Endocytic internalization of Mrp2 to the pericanalicular region was apparent 20 min after a single E(2)17G administration; however, Mrp2 was found more deeply internalized and partially redistributed to the basolateral membrane under sustained cholestasis. In conclusion, acute E(2)17G-induced cholestasis increased permeability of the tight junction, while sustained cholestasis provoked a significant redistribution of ZO-1, occludin, and Mrp2 in addition to increased permeability of the tight junction. Altered tight junction integrity likely contributes to impaired bile secretion and may be causally related to changes in Mrp2 localization. PMID- 17463181 TI - Leptin regulates gallbladder genes related to absorption and secretion. AB - Dysregulation of gallbladder ion and water absorption and/or secretion has been linked to cholesterol crystal and gallstone formation. We have recently demonstrated that obese, leptin-deficient (Lep(ob)) mice have enlarged gallbladder volumes and decreased gallbladder contractility and that leptin administration to these mice normalizes gallbladder function. However, the effect of leptin on gallbladder absorption/secretion is not known. Therefore, we sought to determine whether leptin would alter the expression of genes involved in water and ion transport across the gallbladder epithelium. Affymetrix oligonucleotide microarrays representing 39,000 transcripts were used to compare gallbladder gene expression profiles from 12-wk-old control saline-treated Lep(ob) and from leptin treated Lep(ob) female mice. Leptin administration to Lep(ob) mice decreased gallbladder volume, bile sodium concentration, and pH. Leptin repletion upregulated the expression of aquaporin 1 water channel by 1.3-fold and downregulated aquaporin 4 by 2.3-fold. A number of genes involved in sodium transport were also influenced by leptin replacement. Epithelial sodium channel alpha and sodium hydrogen exchangers 1 and 3 were moderately downregulated by 2.0 , 1.6-, and 1.3-fold, respectively. Carbonic anhydrase-IV, which plays a role in the acidification of bile, was upregulated 3.7-fold. In addition, a number of inflammatory cytokines that are known to influence gallbladder epithelial cell absorption and secretion were upregulated. Thus leptin, an adipocyte-derived cytokine involved with satiety and energy balance, influences gallbladder bile volume, sodium, and pH as well as multiple inflammatory cytokine genes and genes related to water, sodium, chloride, and bicarbonate transport. PMID- 17463182 TI - Wnt5a secretion stimulated by the extracellular calcium-sensing receptor inhibits defective Wnt signaling in colon cancer cells. AB - To understand the role of the colonic extracellular calcium-sensing receptor (CaSR) in calcium chemoprotection against colon cancer, we activated the CaSR with 5 mM Ca(2+) on HT-29 cells, an adenocarcinoma cell line. High Ca(2+) stimulated the upregulation (as assessed by RT-PCR) and the secretion of Wnt5a (assessed by Western blot), a noncanonical Wnt family member. Inhibiting CaSR activity with a short interfering RNA (siRNA) duplex against the CaSR reduced CaSR protein and prevented the secretion of Wnt5a. Dominant negative CaSR (R185Q) or siRNA blocked the high Ca(2+)-mediated inhibition of the beta-catenin reporter TOPflash. The CaSR/Wnt5a inhibition of beta-catenin reporter was prevented by dominant negative ubiquitin ligase seven in absentia homolog 2 (Siah2). In low calcium medium, overexpressing Wnt5a increased Siah2 amplicons and protein. Inducing the expression of full-length adenomatous polyposis coli (APC) prevented CaSRmediated increases of Siah2 and Wnt5a. Overexpressing the receptor tyrosine kinase-like orphan receptor 2 (Ror2) increased Wnt5a and CaSR-mediated inhibition of TOPflash. Conditioned medium from Wnt5a-transfected cells added to HT-29 cells in low-Ca(2+) medium inhibited the beta-catenin reporter. This inhibition was blocked dose responsively by Frizzled-8/Fc chimeric antibody. Overexpression of Ror2 in HT-29 cells in low-Ca(2+) medium increased the inhibition of beta-catenin reporter caused by recombinant Wnt5a protein compared with addition of Wnt5a protein alone. Our findings demonstrate that APC status plays a key role as a determinant of Wnt5a secretion and suggest that CaSR-mediated secretion of Wnt5a will inhibit defective Wnt signaling in APC-truncated cells in an autocrine manner. PMID- 17463183 TI - Pathogenic angiogenesis in IBD and experimental colitis: new ideas and therapeutic avenues. AB - Angiogenesis is now understood to play a major role in the pathology of chronic inflammatory diseases and is indicated to exacerbate disease pathology. Recent evidence shows that angiogenesis is crucial during inflammatory bowel disease (IBD) and in experimental models of colitis. Examination of the relationship between angiogenesis and inflammation in experimental colitis shows that initiating factors for these responses simultaneously increase as disease progresses and correlate in magnitude. Recent studies show that inhibition of the inflammatory response attenuates angiogenesis to a similar degree and, importantly, that inhibition of angiogenesis does the same to inflammation. Recent data provide evidence that differential regulation of the angiogenic mediators involved in IBD-associated chronic inflammation is the root of this pathological angiogenesis. Many factors are involved in this phenomenon, including growth factors/cytokines, chemokines, adhesion molecules, integrins, matrix-associated molecules, and signaling targets. These factors are produced by various vascular, inflammatory, and immune cell types that are involved in IBD pathology. Moreover, recent studies provide evidence that antiangiogenic therapy is a novel and effective approach for IBD treatment. Here we review the role of pathological angiogenesis during IBD and experimental colitis and discuss the therapeutic avenues this recent knowledge has revealed. PMID- 17463184 TI - Helicobacter and gastrin stimulate Reg1 expression in gastric epithelial cells through distinct promoter elements. AB - The gastric pathogen Helicobacter pylori accelerates the progression to gastric cancer but the precise mechanisms that mediate carcinogenesis remain unidentified. We now describe how Helicobacter and gastrin stimulate the expression of a putative growth factor, Reg1, in primary gastric epithelial cells. RT-PCR and Western immunoblotting of human gastric corpus and antrum showed significantly increased Reg1alpha in H. pylori-infected patients. Similarly, Reg1 was increased in the stomachs of H. felis-infected INS-GAS mice. To study transcriptional regulation of the Reg1 gene, we transfected primary mouse gastric glands with -2111 bp and -104 bp Reg1 promoter-luciferase reporter constructs. Expression of both constructs was detected in pepsinogen- and VMAT-2 expressing cells, which corresponds to the normal pattern of expression of human and mouse endogenous Reg1. The expression of both constructs was increased in response to gastrin and H. pylori, and there were potentiating interactions between them; in contrast, only the -2111 bp construct responded to H. felis. Mutation of a C-rich putative regulatory element within the -104 bp sequence abolished the response to gastrin but not to H. pylori whereas mutation of the proximal -98 to -93 region of the promoter reduced the response to H. pylori but not to gastrin. Stimulation of Reg1 by H. pylori required the virulence factor CagA. These data indicate that expression of the putative growth factor Reg1 is controlled through separate promoter elements by gastrin and Helicobacter. PMID- 17463185 TI - PPAR-gamma knockout in pancreatic epithelial cells abolishes the inhibitory effect of rosiglitazone on caerulein-induced acute pancreatitis. AB - Peroxisome proliferator-activated receptor-gamma (PPAR-gamma) agonists, such as the thiazolidinediones (TZDs), decrease acute inflammation in both pancreatic cell lines and mouse models of acute pancreatitis. Since PPAR-gamma agonists have been shown to exert some of their actions independent of PPAR-gamma, the role of PPAR-gamma in pancreatic inflammation has not been directly tested. Furthermore, the differential role of PPAR-gamma in endodermal derivatives (acini, ductal cells, and islets) as opposed to the endothelial or inflammatory cells is unknown. To determine whether the effects of a TZD, rosiglitazone, on caerulein induced acute pancreatitis are dependent on PPAR-gamma in the endodermal derivatives, we created a cell-type specific knock out of PPAR-gamma in pancreatic acini, ducts, and islets. PPAR-gamma knockout animals show a greater response in some inflammatory genes after caerulein challenge. The anti inflammatory effect of rosiglitazone on edema, macrophage infiltration, and expression of the proinflammatory cytokines is significantly decreased in pancreata of the knockout animals compared with control animals. However, rosiglitazone retains its effect in the lungs of the pancreatic-specific PPAR gamma knockout animals, likely due to direct anti-inflammatory effect on lung parenchyma. These data show that the PPAR-gamma in the pancreatic epithelia and islets is important in suppressing inflammation and is required for the anti inflammatory effects of TZDs in acute pancreatitis. PMID- 17463186 TI - Apathy and agitation: challenges and future directions. PMID- 17463187 TI - Epidemiology of apathy in older adults: the Cache County Study. AB - OBJECTIVES: The objectives of this study are to describe the distribution of apathy in community-based older adults and to investigate its relationships with cognition and day-to-day functioning. METHODS: Data from the Cache County Study on Memory, Health and Aging were used to estimate the frequency of apathy in groups of elders defined by demographic, cognitive, and functional status and to examine the associations of apathy with impairments of cognition and day-to-day functioning. RESULTS: Apathy was measured with the Neuropsychiatric Inventory. Clinical apathy (Neuropsychiatric Inventory score > or = 4) was found in 1.4% of individuals classified as cognitively normal, 3.1% of those with a mild cognitive syndrome, and 17.3% of those with dementia. Apathy status was associated with cognitive and functional impairments and higher levels of stress experienced by caregivers. Among participants with normal cognition, apathy was associated with worse performance on the Mini-Mental State Examination, the Boston Naming and Animal Fluency tests, and the Trail Making Test-Part B. The association of apathy with cognitive impairment was independent of its association with Neuropsychiatric Inventory depression. CONCLUSIONS: In a cohort of community based older adults, the frequency and severity of apathy is positively correlated with the severity of cognitive impairment. In addition, apathy is associated with cognitive and functional impairments in elders adjudged to have normal cognition. The results suggest that apathy is an early sign of cognitive decline and that delineating phenotypes in which apathy and a mild cognitive syndrome co-occur may facilitate earlier identification of individuals at risk for dementia. PMID- 17463188 TI - Development of a short version of the Apathy Evaluation Scale specifically adapted for demented nursing home residents. AB - OBJECTIVE: Apathy is among the most frequent neuropsychiatric symptoms in dementia, particularly Alzheimer disease. The Apathy Evaluation Scale (AES) has been widely employed for assessing apathy in different patient groups. To further facilitate the usage of the AES, an abbreviated version was constructed. METHOD: On basis of a sample of 356 nursing home residents, a cross-validation procedure was carried out to develop a brief version of the AES. According to a thorough clinical examination, 85% of the residents were demented, 8% presented with mild cognitive impairment, whereas 7% did not present any cognitive deficits. After subdividing the patient group into two matched samples, the first subsample was used to identify problematic items due to defined psychometric and content related criteria. The original 18-item scale was thus reduced to 10 items. Psychometric properties of the shortened version were subsequently reassessed in the second subsample. RESULTS: The short version demonstrated favorable psychometric properties that could be confirmed by cross-validation with the second sample. Correlations with the original full-length version were high (r = 0.97 for both subsamples); the shortened scale yielded no substantial losses regarding internal consistency or construct validity (correlations with the respective subscales of the Neuropsychiatric Inventory). CONCLUSION: The frequency of apathetic symptoms in the nursing home residents included confirms the clinical importance of apathy for understanding dementia. Given this specific patient population, setting, and mode of data collection, the short-version AES seems to be a valuable and time-efficient instrument for assessing apathy. PMID- 17463189 TI - Neuroanatomical characteristics of geriatric apathy and depression: a magnetic resonance imaging study. AB - OBJECTIVE: Apathy is one of the most common late-life neuropsychiatric syndromes. The objective of our study was to examine the neuroanatomical correlates of apathy in older subjects with and without geriatric major depression (MDD). METHODS: Eighty-four subjects (43 patients with MDD and 41 normal comparison subjects) underwent comprehensive neuropsychiatric examination, physical examination, and high-resolution magnetic resonance imaging (MRI) brain scans on a 1.5T GE MRI scanner. Apathy was assessed using the Apathy Evaluation Scale. MRI image analyses included cortical surface extraction, tissue segmentation, and cortical parcellation methods to measure the gray and white matter volumes in two prefrontal subregions: the anterior cingulate and orbitofrontal cortex. RESULTS: The depressed group had smaller orbitofrontal gray matter volumes compared to the age-matched normal comparison group. The severity of apathy was associated with the decreased gray matter volume in the right anterior cingulate gray matter volumes using partial correlation and regression analyses after controlling for age, sex, and diagnosis. CONCLUSION: Apathy and depression were associated with different anatomical correlates in the prefrontal regions implicated in the regulation of cognition and emotion. Our findings offer new understanding of the neuroanatomical characteristics of apathy and depression in late life, and have broad implications for the neurobiology of behavior. PMID- 17463190 TI - Temporal patterns of agitation in dementia. AB - OBJECTIVE: This study investigated the temporal pattern of agitated behaviors in nursing home residents with dementia and compared it with temporal patterns of pleasure, interest in the environment, and sleep. METHODS: Trained research assistants recorded direct observations of agitation and affect by the Agitation Behavior Mapping Instrument and the Lawton's Modified Behavior Stream for 174 participants from 12 nursing homes. RESULTS: The temporal pattern found for total agitation showed a gradual increase from the morning until approximately 4 pm with a decrease thereafter. Only 26% of the sample showed a clear increase of their agitation in the later hours in comparison to the morning hours based on our criterion. Pleasure did not show a consistent pattern, whereas levels of interest peaked during lunch and dinner times. CONCLUSIONS: The results fit with a hypothesis of fatigue of daytime nursing staff members and disruption during shift change and may also be partially accounted for by resident fatigue. Although the literature supports the notion that there is a peak in overall agitation around 4 pm, the term "sundowning" is not supported and should be replaced. The phenomenon of increased levels of agitation during later hours characterizes only a minority of agitated nursing home residents, albeit the most highly agitated. PMID- 17463191 TI - Neither atypical nor conventional antipsychotics increase mortality or hospital admissions among elderly patients with dementia: a two-year prospective study. AB - BACKGROUND: Antipsychotics are widely used to manage behavioral disorders in patients with dementia. Recently, serious concerns have been raised about the stroke and mortality risk of atypical antipsychotics when administered to patients with dementia. AIM: The aim of this study was to examine the impact of atypical and conventional antipsychotics on mortality and hospital admissions among Finnish elderly institutionalized patients with dementia in a two-year follow up and to compare their prognosis with that of nonusers. PATIENTS AND METHODS: The authors examined 254 very frail patients with dementia, mean age 86 years, from seven Finnish nursing homes and two hospitals in 1999-2000. Medical records provided information on the use of daily antipsychotic medication; central registers confirmed mortality for up to two years. RESULTS: Nearly one half (48.4%) of the patients used antipsychotic medication: 37.4% received conventional neuroleptics (N = 95) and 11.0% received atypical antipsychotics (N = 28). The mean number of hospital admissions was higher among the nonusers than among the users of conventional or atypical antipsychotics. Of the users of atypical antipsychotics (risperidone, olanzapine), 32.1% died within 2 years. The respective figures for users of conventional neuroleptics were 45.3%, and for the nonusers, 49.6%. In the Cox proportional hazard model, a high number of medications and the use of physical restraint predicted higher mortality at two years. The use of atypical antipsychotics showed lower risk of mortality, if any. The respective test for conventional antipsychotics was nonsignificant. CONCLUSION: Among these frail and very old patients with dementia, neither the use of atypical antipsychotics nor the use of conventional neuroleptics increased mortality or hospital admissions. The use of restraints, however, doubled the risk of mortality. PMID- 17463192 TI - Impact of in-home behavioral management versus telephone support to reduce depressive symptoms and perceived stress in Chinese caregivers: results of a pilot study. AB - OBJECTIVE: Recent work has shown that Chinese Americans caring for a family member with dementia experience considerable psychological distress. However, few studies evaluate treatments for them. This study evaluated the efficacy of in home intervention, based on cognitive behavior therapy principles, to relieve stress and depression in female Chinese American caregivers (CGs). METHODS: Fifty five CGs who met inclusion criteria were randomly assigned to a telephone support condition (TSC) or to an in-home behavioral management program (IHBMP) for 4 months. In the TSC, biweekly calls were made and relevant material was mailed. In the IHBMP, specific psychological skills were taught to deal with caregiving stress. CGs were assessed before and after treatment. Outcome measures evaluated overall perceived stress, caregiving-specific stress, and depressive symptoms. RESULTS: CGs in IHBMP were less bothered by caregiving-specific stressors and had lower depression levels than CGs in TSC. There was no difference in overall stress. CGs with low baseline level of self-efficacy for obtaining respite benefited from IHBMP, but showed little improvement in the TSC. CGs with higher self-efficacy benefited from both treatments. CONCLUSION: This intervention is promising and warrants replication in future studies. Additional research is needed to evaluate longer-term effects and to identify individual differences associated with improvement. PMID- 17463193 TI - Aggressive behavior and neuroleptic medication are associated with increased number of alpha1-adrenoceptors in patients with Alzheimer disease. AB - OBJECTIVE: Aggressive behavior in dementia is a major clinical management problem. METHOD: Postmortem brain tissue was obtained from 24 patients with Alzheimer disease (AD) and 25 comparison cases. [3H] Prazosin binding to alpha1 AdR was determined. RESULTS: Aggressive behavior was significantly correlated with alpha1-adrenoceptor number in patients with AD (R(s)=0.454, N=24). Furthermore, patients receiving ongoing neuroleptics had significantly higher Bmax for [3H] prazosin (21 +/- 2, N=9) than those who were not (16 +/- 1, N=15). CONCLUSIONS: Upregulation of alpha1-AdR is associated with aggressive behavior and chronic treatment with neuroleptic medication. PMID- 17463194 TI - Depression and its relationship to function and medical status, by dementia status, in nursing home admissions. AB - OBJECTIVE: To determine rates of depression by dementia status in a statewide sample of nursing home admissions, and associations with medical comorbidity and physical functioning. METHODS: Trained interviewers obtained information from nursing home residents, staff, significant others, and medical records. RESULTS: A total of 22.3% were classified depressed in the nondemented status and 23.6% in the demented status. Depression status was significantly associated with more physical dependencies regardless of dementia status. In the nondemented, there was also a significant positive association with number of comorbidities. One interaction, dementia with comorbidity at the highest levels of comorbidity, was significant in looking at association with depression. CONCLUSION: There is significant depressive symptomatology in nursing home admissions, which is also associated with difficulty in physical function and with the number of medical comorbidities in the nondemented. Application of the two measures used in this study represents a strategy to assess depression in all nursing home residents. PMID- 17463195 TI - E-care: a telecommunications technology intervention for family caregivers of dementia patients. AB - OBJECTIVE: This study evaluated the effectiveness of a technology-based psychoeducational intervention for family caregivers of dementia patients. An additional objective was to determine if the intervention could be implemented by a community-based social service agency. METHODS: Forty-six caregivers were randomly assigned to either a technology-based intervention or an information only control condition. RESULTS: Caregivers assigned to the intervention condition reported a significant decrease in burden postintervention and those who evidenced high depression at baseline experienced a significant decline in depression. CONCLUSION: This study provides evidence that technology offers a cost-effective and practical method for delivering interventions to caregivers. PMID- 17463196 TI - Adjunctive transdermal nicotine reduced behavioral agitation in severe dementia. PMID- 17463197 TI - Characterization of endoleak following endovascular repair of abdominal aortic aneurysms. AB - Aneurysm models have been developed to study the pathobiology of abdominal aortic aneurysm and to evaluate the efficacy of endovascular therapy. The purpose of this review is to describe the use and limitations of current animal and experimental models for the characterization of endoleak following endovascular repair of abdominal aortic aneurysms. PMID- 17463198 TI - Is incidental renal arteriography justified in a population of patients with symptomatic peripheral arterial disease? AB - Renal artery stenosis is a consequence of generalized atherosclerosis and many specialists perform routine selective renal angiography to detect and treat renal artery stenosis. The incidence of clinically important renal artery stenosis is not well defined in patients with symptomatic peripheral arterial disease. The purpose of this study was to better delineate the incidence of and the risk factors associated with renal artery stenosis, renovascular hypertension, and ischemic nephropathy incidentally discovered during angiography for symptomatic peripheral arterial disease. Two hundred consecutive patients undergoing angiographic evaluation of symptomatic lower extremity peripheral arterial disease were studied retrospectively. Angiograms were reviewed for the presence of renal artery stenosis (defined as >or= 25% diameter reduction in either renal artery) and findings were then correlated to the clinical diagnosis of renovascular hypertension (> 50% renal artery stenosis and >or= 3-drug resistive hypertension) and ischemic nephropathy (defined as > 50% bilateral renal artery stenosis, 3-drug hypertension, and creatinine >or= 1.5). Angiographic findings were also correlated with risk factors to determine if a relationship correlated to the presence of and degree of renal artery stenosis. Data were analyzed using the Student's t test, Chi-square model, and multiple logistic regression analysis. The overall incidence of any degree of renal artery stenosis in this study population was 26% (52 patients). Only 24 (12%) patients had an incidental finding of >or= 50% stenosis in either renal artery. Six (3%) of these patients were found to have associated renovascular hypertension. Additionally, 9 (4.5%) patients had coexistent renal insufficiency and significant renal artery stenosis; five with end-stage renal disease on chronic hemodialysis. Only one patient with end-stage renal disease had poorly controlled 3-drug hypertension. Thus definitive ischemic nephropathy was present in only one (0.5%) patient. Statistically significant risk factors associated with the presence of renal artery stenosis include hypertension (P < .001), coronary disease (P = .024), female gender (P = .010), diabetes (P = .039), aorto-iliac disease (P = .031), multiple levels of peripheral arterial disease (P < .001), and age over 60 ( P < .001). While the incidence of renal artery stenosis in patients being evaluated for symptomatic peripheral arterial disease is similar to that reported in the cardiology literature, the incidence of renovascular hypertension and ischemic nephropathy is exceedingly low (3% and 0.5%, respectively)-findings similar to data reported in the general hypertensive population. These data suggest that incidental selective renal angiography is not justified in patients with symptomatic peripheral arterial disease. PMID- 17463199 TI - Determination of functional outcome following upper extremity arterial trauma. AB - Upper extremity arterial trauma may lead to significant disability with a poor functional outcome. This study represents a retrospective review of all trauma patients presenting to a university-affiliated medical center. Patients suffering from upper extremity arterial injuries requiring treatment were identified. The injured vessels were identified along with the mechanism of injury and method of repair. The degree of functional disability was evaluated by using a previously validated questionnaire, the Disabilities of the Arm, Shoulder, and Hand (DASH) Outcome Measure. Between September 1999 and December 2004, 17 patients presented with traumatic arterial injury to the upper extremity, with 9 and 8 patients suffering from blunt and penetrating traumas, respectively. One patient required amputation representing a limb salvage rate of 94%. The mean length of hospitalization was significantly shorter for penetrating trauma (5.1 vs 12 days, P = .03), with blunt trauma victims being more prone to coexisting orthopedic injuries (P = .009). Length of follow-up did not differ between the 2 groups and ranged from 1-60 months. Patients with blunt trauma tended, although not statistically significant, to have higher DASH scores (61.8 vs 22.8, P = .08), indicating a greater degree of disability. By utilizing a validated disability questionnaire, this study confirms that patients suffering from blunt injuries to upper extremity arteries are more likely to have greater degrees of disability affecting everyday activities. PMID- 17463200 TI - Comparison of computed tomography angiography to contrast arteriography for patients undergoing evaluation for lower extremity revascularization. AB - In an effort to explore alternatives to contrast arteriography, we compared computed tomography angiography to contrast arteriography for defining anatomic features of patients undergoing lower extremity revascularization. From November 2003 to March 2004, 36 inpatients with chronic lower extremity ischemia underwent contrast arteriography and computed tomography angiography before undergoing lower extremity revascularization procedures. A Siemens 16 slice multiplanar computed tomography device with bolus tracking was used for these exams. The reports of these tests and images were compared prospectively, and the differences in the aorto-iliac segment, femoral-popliteal, and infrapopliteal segments were noted. The vessels were classified as mild disease (<50%), moderate disease (50%-70%), severe (71%-99%), and occluded. The studies and treatment plans based on these data were compared. The mean age was 76 +/- 12 years (SD). Indications for the procedures included gangrene (45%), ischemic ulcer (32%), rest pain (19%), and severe claudication (3%); 69% were diabetics. Accuracy of computed tomography angiography in the aorto-iliac, femoral-popliteal, and infrapopliteal segments was 100%, 81%, and 59%, respectively. Thirteen of 18 (72%) of these disagreements resulted in a different procedure than that suggested by computed tomography angiography. A review of the data obtained in this series indicated that computed tomography angiography appears to be unable to obtain adequate information in this highly selected population at our institution. PMID- 17463201 TI - Thrombus and calcium in aortic aneurysm necks: validation of a scoring system in a Dutch cohort study. AB - This study was conducted to validate a proposed classification system on the characteristics of the abdominal aortic aneurysm neck. The cohort comprised 100 consecutive patients who underwent open or endovascular aneurysm repair. Aneurysm neck characteristics of diameter, calcium, thrombus, and angulation were reviewed. The presence of calcium at the aortic aneurysm wall was associated with lower body mass index. Thrombus was found in 52% of the patients. Hypertension was correlated with the presence of aortic wall thrombus. At the renal artery level, angulation was anterior in 56%, right lateral in 39%, and left lateral in 27%. At the origin of the aneurysm, angulation was posterior in 76%, right lateral in 46%, and left lateral in 42%. A wide variety in these characteristics was found. A scoring system allows comparison of patient characteristics in studies describing the clinical outcome of endovascular aneurysm repair and should be included in study reports. PMID- 17463202 TI - Modular branched endograft system for aortic aneurysm repair: evaluation in a human cadaver circulation model. AB - A circulation model was created in 6 nonaneurysmal human cadavers to evaluate the deliverability, deployment, and acute performance of a modular branched endograft system for treatment of aortic aneurysms containing essential branch vessels. Two fenestrations were created in an appropriately sized aortic main endograft. Under fluoroscopic guidance, the main endograft was advanced to the target site and the fenestrations were aligned with the ostia of the renal arteries. Branch grafts were placed through the fenestrations into the renal arteries. The outcome was evaluated by post implant angiography and autopsy. Eleven branch grafts were deployed at the target site. All targeted renal arteries showed good patency. At autopsy, all main endografts were adequately deployed, and 10 of 11 branch grafts were locked in place. In this model, deliverability and deployment of the modular branch graft system is feasible in a reliable, predictable, and timely fashion. PMID- 17463203 TI - Recanalization of concomitant iliac artery and common femoral artery occlusions using an antegrade hybrid (open/endovascular) approach. AB - The endovascular treatment of patients with bulky iliofemoral occlusions usually requires a femoral endarterectomy, coupled with iliac recanalization. This requires crossing the occlusions with a guidewire, which is usually attempted in a retrograde fashion. If this fails, then a surgical inflow procedure is necessary. Antegrade crossing of an iliac occlusion either from the contralateral femoral or transbrachial approach and retrieval of the guidewire during the ensuing femoral endarterectomy obviates the need for luminal reentry, ensures inflow by endovascular recanalization of the iliac artery, and avoids the need for surgical bypass. This underused technique should be considered in such situations, and its details are described. PMID- 17463204 TI - Large abdominal aortic aneurysm with aortocaval fistula. PMID- 17463205 TI - Systemic thrombolysis using tissue plasminogen activator for a patient with paradoxic embolism: a case report. AB - When thrombotic material that originates from deep venous thrombosis of the lower extremities is washed out into the pulmonary vasculature, pulmonary embolization occurs. Pulmonary embolism and associated acute peripheral ischemia suggest the diagnosis of paradoxic embolism, which is most often associated with a patent foramen ovale. Therapeutic options include anticoagulation, mechanical/chemical thrombus dissolution, inferior vena cava filtration, and closure of the intracardiac defect. The diagnosis and treatment are described of an elderly female who presented with lower extremity deep venous thrombosis and massive pulmonary embolism complicated by paradoxic emboli to the left subclavian artery as well as the celiac artery. PMID- 17463206 TI - Feasibility of renal artery stenting before, during, and after endovascular aneurysm repair with suprarenal fixation endograft. AB - Suprarenal fixation by means of proximal bare stent may help prevent endograft migration and proximal endoleak. It seems not to compromise renal perfusion and function; however, it is still unclear whether its presence is in conflict with stenting of the renal arteries to treat associated renal stenosis. We report five cases of renal artery stenting performed before, at the same time, or after endovascular treatment of abdominal aortic aneurysm with a suprarenal fixation endograft (Zenith; Cook, Biaverskov, Denmark). Suprarenal fixation endograft seems not to affect renal artery stenting regardless the timing of the procedure. PMID- 17463207 TI - Iliac conduit for deploying aortic endografts: an all-autogenous option. AB - Unfavorable iliac anatomy, such as small caliber or circumferential calcification, can cause problems during delivery of aortic endografts and lead to potential complications. An iliac artery conduit "chimney" has been used to deliver the large caliber endoprosthesis using a Dacron graft sewn onto the iliac bifurcation. An all-autogenous revascularization option allows for hypogastric bypass while limiting the use of synthetic graft. PMID- 17463208 TI - Spiral saphenous vein graft for portal vein reconstruction in pancreatic cancer surgery. AB - The curative strategy for most pancreatic cancer is surgical resection. Extensive resection with lymph node dissection is the key to providing long-term survival. However, early diagnosis of pancreatic cancer is not always possible (ie, resectability is limited). One reason for such a nonresectable condition is vascular invasion or encasement. Portal vein involvement has been a contraindication for pancreatic cancer surgery for most general surgeons. Combining oncologic and vascular surgeons in the procedure has been a good solution. A multidisciplinary approach that includes general and vascular surgeons is appropriate in selected patients requiring vascular reconstruction at the time of pancreatectomy. The objective of this paper is to report a case in which spiral saphenous vein was used for portal vein reconstruction during pancreatic cancer resection. PMID- 17463209 TI - Stent graft repair of traumatic vertebral pseudoaneurysm with arteriovenous fistula. AB - Traumatic vertebral pseudoaneurysm is a rare occurrence. Such case with an enlarging symptomatic vertebral arteriovenous (AV) fistula was successfully treated with a covered endostent. Presentation, diagnosis and different acceptable treatment plans to this condition are further discussed. Stentgrafts can play an important and effective role in therapy of traumatic vertebral pseudoaneurysm with a concomitant AV fistula. PMID- 17463210 TI - Evolution of bacterial arteritis into a mycotic aortic aneurysm. AB - Arteritis and mycotic aneurysms have been well described for more than 100 years. The authors report a case of bacterial arteritis that presented with pneumatosis of the aortic wall and that evolved over 1 week into an infected abdominal aortic aneurysm. This case documents the rapid progression from arteritis to mycotic aneurysm, highlighting the need for close radiologic follow-up and aggressive medical and surgical management. PMID- 17463211 TI - Endovascular stent exclusion of a hepatic artery pseudoaneurysm. AB - Presented is a patient with an iatrogenic hepatic artery pseudoaneurysm that was treated by endovascular stent grafting. Endovascular stent grafting of a hepatic artery pseudoaneurysm offers a safe and potentially less morbid alternative to an open repair. The report stresses the necessity of careful preoperative evaluation with angiography to determine the feasibility of the procedure. An aggressive approach to treating hepatic artery pseudoaneurysms is advocated because of the poor correlation between size and their tendency to rupture. With an increasing rate of diagnosis of visceral artery aneurysms in elderly, debilitated patients, endovascular repair is anticipated to have an increasing role and should be considered a first-line therapy in anatomically suitable candidates. PMID- 17463212 TI - Can clopidogrel monotherapy influence vascular remodeling? PMID- 17463213 TI - Effect of aprepitant on the pharmacokinetics of intravenous midazolam. AB - Oral aprepitant 125 mg, an antiemetic and a moderate inhibitor of the metabolism of oral midazolam, was assessed for interaction with intravenous midazolam in 12 subjects randomized to intravenous midazolam 2 mg +/- oral aprepitant 125 mg. The hypothesis was that midazolam AUC would not change by more than 2-fold (consistent with no more than weak inhibition) when midazolam + aprepitant was compared with midazolam alone. An AUC geometric mean ratio (midazolam + aprepitant/midazolam) with 90% confidence interval upper bound < or =2.0 (an increase in midazolam felt to be of modest clinical significance in the highly monitored perioperative period) was prespecified. Aprepitant increased intravenous midazolam AUC(0-infinity) 1.47-fold (90% confidence interval, 1.36 1.59), which fell within the prespecified criterion. PMID- 17463214 TI - Effect of the rate of niacin administration on the plasma and urine pharmacokinetics of niacin and its metabolites. AB - The metabolic profile of niacin is influenced by the rate of niacin administration. This study characterizes the effect of administration rate on the pharmacokinetics of niacin and its metabolites. Twelve healthy males were enrolled in an open-label, dose-rate escalation study and received 2000 mg niacin at 3 different dosing rates. Plasma was analyzed for niacin, nicotinuric acid, nicotinamide, and nicotinamide-N-oxide. Urine was analyzed for niacin and the metabolites nicotinuric acid, nicotinamide, nicotinamide-N-oxide, N methylnicotinamide, and N-methyl-2-pyridone-5-carboxamide. C(max) and AUC(0-t) for niacin and nicotinuric acid increased with an increase in dosing rate. The changes observed in plasma nicotinamide and nicotinamide-N-oxide parameters, however, did not correlate to dosing rate. The total amount of niacin and metabolites excreted in urine was comparable for all 3 treatments. However, with the increase in dosing rate, urine recovery of niacin and nicotinuric acid showed a significant increase, whereas N-methyl-2-pyridone-5-carboxamide and N methylnicotinamide showed a significant decrease. PMID- 17463215 TI - Replicate study design in bioequivalency assessment, pros and cons: bioavailabilities of the antidiabetic drugs pioglitazone and glimepiride present in a fixed-dose combination formulation. AB - An open-label, randomized, 2-sequence, 4-period crossover (7-day washout period between treatment), replicate design study was conducted in 37 healthy subjects to assess intersubject and intrasubject variabilities in the peak (Cmax) and total (AUC) exposures to 2 oral antidiabetic drugs, pioglitazone and glimepiride, after single doses of 30 mg pioglitazone and 4 mg glimepiride, given under fasted state, as commercial tablets coadministered or as a single fixed-dose combination tablet. Variabilities for AUC(infinity) for coadministered and fixed-dose combination treatments were similar: 16% to 19% (intra) and 23% to 25% (inter) for pioglitazone and 18% to 19% (intra) and 29% to 30% for glimepiride (inter, excluding 1 poor metabolizer). Fixed-dose combination/coadministered least squares mean ratios of >or=0.86 and the 90% confidence intervals of these ratios for pioglitazone and glimepiride of between 0.80 and 1.25 for Cmax, AUC(lqc), and AUC(infinity) met the bioequivalency standards. Gender analysis showed that women showed mean of 16% and 30% higher exposure than men for glimepiride (excluding 1 poor metabolizer) and pioglitazone, respectively. There was considerable overlapping in the AUC(infinity) values, making gender-dependent dosing unnecessary. Patients taking pioglitazone and glimepiride as cotherapy may replace their medication with a single fixed-dose combination tablet containing these 2 oral antidiabetic drugs. PMID- 17463216 TI - Effective integration of systems biology, biomarkers, biosimulation, and modeling in streamlining drug development. PMID- 17463217 TI - Coadministration of estradiol/drospirenone and indomethacin does not cause hyperkalemia in healthy postmenopausal women: a randomized open-label crossover study. AB - The effect of drospirenone on plasma potassium when coadministered with nonsteroidal anti-inflammatory drugs, such as indomethacin, is unknown. An open label crossover study investigated the effects of estradiol/drospirenone and indomethacin coadministration on plasma potassium levels in 32 postmenopausal women. Each participant received 2 treatments in random order: indomethacin alone for 5 days and estradiol/drospirenone alone for 12 days, then estradiol/drospirenone plus indomethacin for 5 days. Plasma potassium profiles (24 hours) were measured on the first and last days of indomethacin administration. No difference was seen between treatments in the area under the curve or maximum concentration of plasma potassium. No participant experienced hyperkalemia (potassium >5.5 mmol/L). Twenty-seven participants had at least 1 potassium value above the upper limit of normal (4.4 mmol/L), but these occurred during both treatments. Coadministration of estradiol/drospirenone and a nonsteroidal anti-inflammatory drug such as indomethacin is not expected to result in increased plasma potassium or hyperkalemia in healthy postmenopausal women. PMID- 17463218 TI - Effect of food on the pharmacokinetics of rifalazil, a novel antibacterial, in healthy male volunteers. AB - Rifalazil is a new antibiotic structurally related to rifampin but devoid of the metabolic liabilities typically associated with the rifamycin class of antibiotics. A randomized, 3-way crossover study in healthy male volunteers (n = 12) investigated the safety and pharmacokinetics of a single 25-mg oral rifalazil dose administered under a standard breakfast containing fat as 30% of calories, a high-fat breakfast containing fat as 60% of calories, and an overnight fast of 10 hours with a 21- to 28-day washout between doses. Systemic exposure to rifalazil based on Cmax, AUC(0-Tlast), and AUC(0-infinity) was increased progressively as the fat content of the test breakfast was increased from 30% to 60% compared with fasting. The confidence intervals for both fat-containing breakfasts are outside the limits of 80% to 125% allowed for food effect bioequivalence based on Cmax, AUC(0-Tlast), and AUC(0-infinity). This food effect may be a result of increased fractional absorption with increasing dietary fat content. Another striking finding was the large reduction of the pharmacokinetic intersubject variability after rifalazil administration with food. Rifalazil was safe and well tolerated under fed and fasted conditions. PMID- 17463219 TI - Pharmacokinetics of an oral drug (acetaminophen) administered at various times relative to subcutaneous injection of pramlintide in subjects with type 2 diabetes. AB - Pramlintide, an adjunct treatment to mealtime insulin for patients with type 2 and type 1 diabetes, aids glycemic control by suppressing postprandial glucagon secretion, slowing gastric emptying, and enhancing satiety. Because gastric emptying affects oral medication absorption, this placebo-controlled, single blind, crossover study examined the absorption of 1000 mg of acetaminophen elixir administered -2, -1, 0, +1, and +2 hours relative to pramlintide (120 microg) or 0 hours relative to placebo in 24 patients with type 2 diabetes. When acetaminophen administration occurred 0, +1, or +2 hours relative to pramlintide, the maximum observed plasma concentration of acetaminophen decreased 14% to 29%, and time to maximum observed plasma concentration increased by 0.8 to 1.2 hours compared with administration 0 hours relative to placebo. Pramlintide treatment slowed but did not alter the extent of acetaminophen absorption (area under the concentration-time curve). No serious adverse events or withdrawals were reported. Oral agents should be administered at least 1 hour before or 2 hours after pramlintide injection if rapid onset of action is required for efficacy. PMID- 17463224 TI - Low-density lipoprotein receptor-related protein promotes amyloid precursor protein trafficking to lipid rafts in the endocytic pathway. AB - The major defining pathological hallmark of Alzheimer's disease (AD) is the accumulation of amyloid beta protein (Abeta), a small peptide derived from beta- and gamma-secretase cleavages of the amyloid precursor protein (APP). Recent studies have shown that beta- and gamma-secretase activities of BACE1 and presenilin, respectively, are concentrated in intracellular lipid raft microdomains. However, the manner in which APP normally traffics to lipid rafts is unknown. In this study, using transient transfection and immuno-precipitation assays, we show that the cytoplasmic domain of low-density lipoprotein receptor related protein (LRP) interacts with APP and increases Abeta secretion and APP beta-CTF (C-terminal fragment) generation by promoting BACE1-APP interaction. We also employed discontinuous sucrose density gradient ultracentrifugation to show that the LRP cytoplasmic domain-mediated effect was accompanied by greatly increased localization of APP and BACE1 to lipid raft membranes, where beta- and gamma-secretase activities are highly enriched. Moreover, we provide evidence that endogenous LRP is required for the normal delivery of APP to lipid rafts and Abeta generation primarily in the endocytic but not secretory pathway. These results may provide novel insights to block Abeta generation by targeting LRP mediated delivery of APP to raft microdomains. PMID- 17463225 TI - Functional characterization, tuning, and regulation of visual pigment gene expression in an anadromous lamprey. AB - Lampreys are one of the two surviving groups of jawless vertebrates, whose ancestors arose more than 540 million years ago. Some species, such as Geotria australis, are anadromous, commencing life as ammocoetes in rivers, migrating downstream to the sea, and migrating back into rivers to spawn. Five photoreceptor types and five retinal cone opsin genes (LWS, SWS1, SWS2, RhA, and RhB) have previously been identified in G. australis. This implies that the ancestral vertebrates possessed photopic or cone-based vision with the potential for pentachromacy. Changes in the morphology of photoreceptors and their spectral sensitivity are encountered during differing aquatic phases of the lamprey lifecycle. To understand the molecular basis for these changes, we characterized the visual pigments and measured the relative levels of opsin expression over two lifecycle phases that are accompanied by contrasting ambient light environments. By expressing recombinant opsins in vitro, we show that SWS1, SWS2, RhA, and RhB visual pigments possess lambda(max) values of 359, 439, 497, and 492 nm respectively. For the LWS visual pigment, we predict a lambda(max) value of 560 nm based on key spectral tuning sites in other vertebrate LWS opsins. Quantitative reverse transcriptase-polymerase chain reaction reveals that the retinal opsin genes of G. australis are differentially regulated such that the visual system switches from a broad sensitivity across a wide spectral range to a much narrower sensitivity centered around 490-500 nm on transition from marine to riverine conditions. These quantitative changes in visual pigment expression throughout the lifecycle may directly result from changes in the lighting conditions of the surrounding milieu. PMID- 17463226 TI - The E3 ligase Aip4/Itch ubiquitinates and targets ErbB-4 for degradation. AB - The ErbB-4 receptors are unique in the EGFR/ErbB family for the ability to associate with WW domain-containing proteins. To identify new ligands of the cytoplasmic tail of ErbB-4, we panned a brain cDNA phage library with ErbB-4 peptides containing sequence motifs corresponding to putative docking sites for class-I WW domains. This approach led to identification of AIP4/Itch, a member of the Nedd4-like family of E3 ubiquitin protein ligases, as a protein that specifically interacts with and ubiquitinates ErbB-4 in vivo. Interaction with the ErbB-4 receptors occurs via the WW domains of AIP4/Itch. Functional analyses demonstrate that AIP4/Itch is recruited to the ErbB-4 receptor to promote its polyubiquitination and degradation, thereby regulating stability of the receptor and access of receptor intracellular domains to the nuclear compartment. These findings expand our understanding of the mechanisms contributing to the integrity of the ErbB signaling network and mechanistically link the cellular ubiquitination pathway of AIP4/Itch to the ErbB-4 receptor. PMID- 17463227 TI - Delivery of short interfering RNA using endosomolytic cell-penetrating peptides. AB - Cell-penetrating peptides (CPPs) are peptides able to promote uptake of various cargos, including proteins and plasmids. Advances in recent years imply the uptake to be endocytic, where the current hurdle for efficient intracellular delivery is material being retained in the endosomes. In this study we wanted to compare the ability of various established CPPs to deliver siRNA and induce gene silencing of luciferase, with a novel designed penetratin analog having endosomolytic properties, using a noncovalent strategy. In principal, the penetratin analog EB1 will, upon protonation in the early-late endosomes, be able to form an amphipathic alpha helix resulting in permeabilization of the endosomal membrane. We demonstrate that even though all CPPs evaluated in this study can form complexes with siRNA, there is not a direct relationship between the complex formation ability and delivery efficacy. More important, although all CPPs significantly promote siRNA uptake, in some cases no gene silencing effect can be observed unless endosomal escape is induced. We find the designed endosomolytic peptide EB1 to be far more effective both in forming complexes and transporting biologically active siRNA than its parent peptide penetratin. We believe that developing CPPs with increased endosomolytical properties is a necessary step toward achieving biological effects at low concentrations for future in vivo applications. PMID- 17463228 TI - Perceptual weighting of stop consonant cues by normal and impaired listeners in reverberation versus noise. AB - PURPOSE: To determine if listeners with normal hearing and listeners with sensorineural hearing loss give different perceptual weightings to cues for stop consonant place of articulation in noise versus reverberation listening conditions. METHOD: Nine listeners with normal hearing (23-28 years of age) and 10 listeners with sensorineural hearing loss (31-79 years of age, median 66 years) participated. The listeners were asked to label the consonantal portion of synthetic CV stimuli as either /p/ or /t/. Two cues were varied: (a) the amplitude of the spectral peak in the F4/F5 frequency region of the burst was varied across a 30-dB range relative to the adjacent vowel peak amplitude in the same frequency region, (b) F2/F3 formant transition onset frequencies were either appropriate for /p/, /t/ or neutral for the labial/alveolar contrast. RESULTS: Weightings of relative amplitude and transition cues for voiceless stop consonants depended on the listening condition (quiet, noise, or reverberation), hearing loss, and age of listener. The effects of age with hearing loss reduced the perceptual integration of cues, particularly in reverberation. The effects of hearing loss reduced the effectiveness of both cues, notably relative amplitude in reverberation. CONCLUSIONS: Reverberation and noise conditions have different perceptual effects. Hearing loss and age may have different, separable effects. PMID- 17463229 TI - The consistency of sentence intelligibility across three types of signal distortion. AB - PURPOSE: To examine the extent to which sentences retain their levels of spoken intelligibility relative to other sentences in a set (the sentence effect) across different types of signal distortion. METHOD: The Central Institute for the Deaf (CID) sentences were rendered difficult to understand through the addition of broadband noise. These intelligibility data were compared with those from previous studies in which the sentences were distorted through filtering and visual-only conditions of speechreading. The extent to which the various sentences retained their intelligibility rankings was examined using an analysis of variance model and by correlating individual sentence means across conditions. RESULTS: The sentences accounted for a large portion of the variance, and individual sentence scores were highly correlated across conditions involving a single distortion type. However, correlations were lower when conditions involving noise were compared with those involving filtering. Surprisingly, correlations across auditory distortions were almost identical to those observed across auditory and visual modalities. These comparisons, reflecting the consistency of sentence difficulty independent of presentation characteristics, accounted for approximately 25% of the variance in sentence-recognition performance. CONCLUSION: There exists a sentence effect that holds across various types of signal distortion, but the strongest form is restricted not only within modalities but within particular forms of distortion. PMID- 17463230 TI - Auditory speech recognition and visual text recognition in younger and older adults: similarities and differences between modalities and the effects of presentation rate. AB - PURPOSE: To examine age-related differences in auditory speech recognition and visual text recognition performance for parallel sets of stimulus materials in the auditory and visual modalities. In addition, the effects of variation in rate of presentation of stimuli in each modality were investigated in each age group. METHOD: A mixed-model design was used in which 3 independent groups (13 young adults with normal hearing, 10 elderly adults with normal hearing, and 16 elderly hearing-impaired adults) listened to auditory speech tests (a sentence-in-noise task, time-compressed monosyllables, and a speeded-spelling task) and viewed visual text-based analogs of the auditory tests. All auditory speech materials were presented so that the amplitude of the speech signal was at least 15 dB above threshold through 4000 Hz. RESULTS: Analyses of the group data revealed that when baseline levels of performance were used as covariates in the group analyses the only significant group difference was that both elderly groups performed worse than the young group on the auditory speeded-speech tasks. Analysis of individual data, using correlations, factor analysis, and linear regression, was generally consistent with the group data and revealed significant, moderate correlations of performance for similar tasks across modalities, but stronger correlations across tasks within a modality. This suggests that performance on these tasks was mediated both by a common underlying factor, such as cognitive processing, as well as modality-specific processing. CONCLUSION: Performance on measures of auditory processing of speech examined here was closely associated with performance on parallel measures of the visual processing of text obtained from the same participants. Young and older adults demonstrated comparable abilities in the use of contextual information in each modality, but older adults, regardless of hearing status, had more difficulty with fast presentation of auditory speech stimuli than young adults. There were no differences among the 3 groups with regard to the effects of presentation rate for the visual recognition of text, at least for the rates of presentation used here. PMID- 17463231 TI - Ranking hearing aid input-output functions for understanding low-, conversational , and high-level speech in multitalker babble. AB - PURPOSE: To determine the rankings of 6 input-output functions for understanding low-level, conversational, and high-level speech in multitalker babble without manipulating volume control for listeners with normal hearing, flat sensorineural hearing loss, and mildly sloping sensorineural hearing loss. METHOD: Peak clipping, compression limiting, and 4 wide dynamic range compression (WDRC) input output functions were compared in a repeated-measure design. Interactions among the compression characteristics were minimized. Speech and babble were processed and recorded at 3 input levels: 45, 65, and 90 dB sound pressure level. Speech recognition of 3 groups of listeners (n = 6/group) was tested for speech processed by each input-output function and at each input level. RESULTS: Input output functions that made low-level speech audible and high-level speech less distorted by avoiding peak clipping or severe compression yielded higher speech recognition scores. These results are consistent with previous findings in the literature. CONCLUSION: WDRCs with the low compression ratio region extended to a high input level or with a high compression limiting threshold were the best for speech recognition in babble when the hearing aid user cannot or does not want to manipulate the volume control. Future studies on subjective preferences of different input-output functions are needed. PMID- 17463232 TI - Speech perception in MRI scanner noise by persons with aphasia. AB - PURPOSE: To examine reductions in performance on auditory tasks by aphasic and neurologically intact individuals as a result of concomitant magnetic resonance imaging (MRI) scanner noise. METHOD: Four tasks together forming a continuum of linguistic complexity were developed. They included complex-tone pitch discrimination, same-different discrimination of minimal pair syllables, lexical decision, and sentence plausibility. Each task was performed by persons with aphasia (PWA) and by controls. The stimuli were presented in silence and also in the noise recorded from within the bore of a 3 Tesla MRI scanner at 3 signal-to noise (S/N) ratios. RESULTS: Across the 4 tasks, the PWA scored lower than the controls, and performance fell as a function of decreased S/N. However, the rate at which performance fell was not different across the 2 listener groups in any task. CONCLUSIONS: Depending on the relative levels of the signals and noise, the intense noise accompanying MRI scanning has the potential to severely disrupt performance. However, PWA are no more susceptible to the disruptive influence of this noise than are unimpaired individuals usually employed as controls. Thus, functional MRI data from aphasic and control individuals may be interpreted without complications associated with large interactions between scanner noise and performance reduction. PMID- 17463233 TI - Training effects on speech production using a hands-free electromyographically controlled electrolarynx. AB - PURPOSE: The electrolarynx (EL) is a widely used device for alaryngeal speech, but it requires manual operation and produces voice that typically has a constant fundamental frequency. An electromyographically controlled EL (EMG-EL) was designed and implemented to provide hands-free control with dynamic pitch modulation. METHOD: Three participants who underwent total laryngectomy surgery and 4 participants with normal voice were trained to produce EMG-EL speech through a multiple-baseline, successive-stage protocol. Baseline performance was established through 3 testing probes, followed by multiple hour-long training sessions. RESULTS: At the end of the training, all participants learned to initiate, sustain, and terminate EMG-EL activation in correspondence with articulation, and most were able to modulate the pitch to produce intonational contrasts. After completing the testing/training protocol, 1 of the 3 participants who underwent total laryngectomy was encouraged to independently use the EMG-EL at his residence. This participant sustained his performance for an additional 6 weeks and also used the EMG-EL successfully to communicate over the phone. CONCLUSIONS: Our findings suggest that some participants with laryngectomies and vocally normal individuals can learn to produce hands-free speech using the EMG-EL device within a few hours and that significant additional gains in device control (particularly pitch modulation) are attainable through subsequent training sessions. PMID- 17463234 TI - Physiologic development of tongue-jaw coordination from childhood to adulthood. AB - PURPOSE: This investigation aimed to examine the development of tongue-jaw coordination during speech from childhood to adolescence. METHOD: Electromagnetic articulography was used to track tongue and jaw motion in 48 children and adults (aged 6-38 years) during productions of /t/ and /k/ embedded in sentences. RESULTS: The coordinative organization of the tongue and jaw exhibited changes until the age of 8-11 years and continued to undergo refinement into late adolescence. The tongue-tip and tongue-body were observed to develop unique kinematic relations with the jaw. While tongue-tip movement became increasingly synchronized with jaw movement, tongue-body and jaw retained movement independence but developed a more consistent kinematic relation. CONCLUSION: The present results support the notion that speech motor development is nonuniform, with a refinement period from mid-childhood to late adolescence. PMID- 17463235 TI - Speaking-related dyspnea in healthy adults. AB - PURPOSE: To reveal the qualities and intensity of speaking-related dyspnea in healthy adults under conditions of high ventilatory drive, in which the behavioral and metabolic control of breathing must compete. METHOD: Eleven adults read aloud while breathing different levels of inspired carbon dioxide (CO(2)). After the highest level, participants provided unguided descriptions of their experiences and then selected descriptors from a list. On a subsequent day, participants read aloud while breathing high CO(2) as before, then rated air hunger, physical exertion, and mental effort (with definitions provided). Recordings were made of ventilation (with respiratory magnetometers), end-tidal partial pressure of CO(2), transcutaneous PCO(2), oxygen saturation, noninvasive blood pressure, heart rate, and the speech signal. RESULTS: Unguided descriptions were found to reflect the qualities of air hunger, physical exertion (work), mental effort, and speech-related observations. As CO(2) stimulus strength increased, participants experienced increased perception of air hunger, physical exertion, and mental effort. Simultaneous increases were observed in ventilation, tidal volume, end-inspiratory and end-expiratory volumes, expiratory flow during speaking, nonlinguistic junctures, and nonspeech expirations. CONCLUSION: Two qualities of speaking-related dyspnea--air hunger and physical exertion--are the same as those reported for many other types of nonspeech dyspnea conditions and, therefore, may share the same physiological mechanisms. The mental effort quality associated with speaking-related dyspnea may reflect a conscious drive to balance speech requirements and ventilatory demands. These findings have implications for developing better ways to evaluate and manage clients with respiratory-based speech problems. PMID- 17463236 TI - Electropalatographic assessment of tongue-to-palate contact patterns and variability in children, adolescents, and adults. AB - PURPOSE: To investigate the developmental time course of tongue-to-palate contact patterns during speech from childhood to adulthood using electropalatography (EPG) and a comprehensive profile of data analysis. METHOD: Tongue-to-palate contacts were recorded during productions of /t/, /l/, /s/, and /k/ in 48 children, adolescents and adults (aged 6-38 years) using the Reading Electropalatograph system. RESULTS: A protracted course of development for lingual control was indicated, with significant changes occurring until age 11 years; the adolescent period was in turn characterized by continual refinement of articulatory control. With maturity, a reduction in the amount of palatal contact and an anterior shift in the place of articulation was evident during anterior consonant productions, whereas the tongue-back-to-palate contact pattern became more consistent for the velar stop /k/. CONCLUSION: These results support that maturation of the speech motor system is nonuniform. PMID- 17463238 TI - Speed of processing, working memory, and language impairment in children. AB - PURPOSE: Children with language impairment (LI) often perform below the level of typically developing peers on measures of both processing speed and working memory. This study examined the relationship between these 2 types of measures and attempted to determine whether such measures can account for the LI itself. METHOD: Fourteen-year-old children with LI and their typically developing peers participated in a wide range of processing speed and working memory tasks and were administered a comprehensive language test battery. Confirmatory factor analyses were used to compare 3 nested models designed to examine the dimensionality of the speed and working memory measures. A model that included a general speed factor was also evaluated. RESULTS: The models meeting our evaluation criteria treated speed and working memory as separable factors. Furthermore, nonverbal as well as verbal processing factors emerged from these analyses. Latent variable regression analyses showed that each of the appropriate models accounted for 62% of the variance in the children's concurrent composite language test scores, with verbal working memory making the largest contribution. CONCLUSIONS: These findings shed light on the relationship among different types of processing and suggest that processing factors can contribute to the understanding of language disorders. PMID- 17463237 TI - Profiles of vocal development in young cochlear implant recipients. AB - PURPOSE: The main purpose of this investigation was to examine the effects of cochlear implant experience on prelinguistic vocal development in young deaf children. Procedure A prospective longitudinal research design was used to document the sequence and time course of vocal development in 7 children who were implanted between 10 and 36 months of age. Speech samples were collected twice before implant activation and on a monthly basis thereafter for up to 2 years. Children's vocalizations were classified according to the levels of the Stark Assessment of Early Vocal Development--Revised (SAEVD-R; S. Nathani, D. J Ertmer, & R. E. Stark, 2006). RESULTS: The main findings were (a) 6 of 7 children made advancements in vocal development after implantation; (b) children implanted between 12 and 36 months progressed through SAEVD-R levels in the predicted sequence, whereas a child implanted at a younger age showed a different sequence; (c) milestones in vocal development were often achieved with fewer months of hearing experience than observed in typically developing infants and appeared to be influenced by age at implantation; and (d) in general, children implanted at younger ages completed vocal development at younger chronological ages than those implanted later in life. Specific indicators of benefit from implant use were also identified. CONCLUSION: The time course of vocal development in young cochlear implant recipients can provide clinically useful information for assessing the benefits of implant experience. Studies of postimplantation vocal development have the potential to inform theories of spoken language development. PMID- 17463239 TI - The preschool repetition test: an evaluation of performance in typically developing and clinically referred children. AB - PURPOSE: To determine the psychometric properties of the Preschool Repetition (PSRep) Test (Roy & Chiat, 2004), to establish the range of performance in typically developing children and variables affecting this performance, and to compare the performance of clinically referred children. METHOD: The PSRep Test comprises 18 words and 18 phonologically matched nonwords systematically varied for length and prosodic structure. This test was administered to a typical sample of children aged 2;0-4;0 (n = 315) and a clinic sample of children aged 2;6-4;0 (n = 168), together with language assessments. RESULTS: Performance in the typical sample was independent of gender and socioeconomic status but was affected by age, item length, and prosodic structure and was moderately correlated with receptive vocabulary. Performance in the clinic sample was significantly poorer but revealed similar effects of length and prosody and similar relations to language measures overall, with some notable exceptions. Test-retest reliability and interrater reliability were high. CONCLUSION: The PSRep Test is a viable and informative test. It differentiates within and between typical and clinic samples of children and reveals some unusual profiles within the clinic sample. These findings lay the foundations for a follow-up study of the clinic sample for investigation of the predictive value of the test. PMID- 17463240 TI - Relations between segmental and motor variability in prosodically complex nonword sequences. AB - PURPOSE: To assess how prosodic prominence and hierarchical foot structure influence segmental and articulatory aspects of speech production, specifically segmental accuracy and variability, and oral movement trajectory variability. METHOD: Thirty individuals participated: 10 young adults, 10 children who are normally developing, and 10 children diagnosed with specific language impairment. Segmental error and segmental variability and movement trajectory variability were compared in low and high prosodic prominence conditions (i.e., strong and weak syllables) and in different prosodic foot structures. RESULTS: Between participants findings were that both groups of children showed more segmental error and segmental variability and more movement trajectory variability than did adults. A similar within-participant pattern of results was observed for all 3 groups. Prosodic prominence influenced both segmental and motor levels of analysis, with weak syllables produced less accurately and with more lip and jaw movement trajectory variability than strong syllables. However, hierarchical foot structure affected segmental but not motor measures of speech production accuracy and variability. CONCLUSIONS: Motor and segmental variables were not consistently aligned. This pattern of results has clinical implications because inferences about motor variability may not directly follow from observations of segmental variability. PMID- 17463241 TI - Exploring the utility of narrative analysis in diagnostic decision making: picture-bound reference, elaboration, and fetal alcohol spectrum disorders. AB - PURPOSE: To evaluate classification accuracy and clinical feasibility of a narrative analysis tool for identifying children with a fetal alcohol spectrum disorder (FASD). METHOD: Picture-elicited narratives generated by 16 age-matched pairs of school-aged children (FASD vs. typical development [TD]) were coded for semantic elaboration and reference strategy by judges who were unaware of age, gender, and group membership of the participants. Receiver operating characteristic (ROC) curves were used to examine the classification accuracy of the resulting set of narrative measures for making 2 classifications: (a) for the 16 children diagnosed with FASD, low performance (n = 7) versus average performance (n = 9) on a standardized expressive language task and (b) FASD (n = 16) versus TD (n = 16). RESULTS: Combining the rates of semantic elaboration and pragmatically inappropriate reference perfectly matched a classification based on performance on the standardized language task. More importantly, the rate of ambiguous nominal reference was highly accurate in classifying children with an FASD regardless of their performance on the standardized language task (area under the ROC curve = .863, confidence interval = .736-.991). CONCLUSION: Results support further study of the diagnostic utility of narrative analysis using discourse level measures of elaboration and children's strategic use of reference. PMID- 17463242 TI - Discourse skills of boys with fragile X syndrome in comparison to boys with Down syndrome. AB - PURPOSE: This study compared the conversational discourse skills of boys who have fragile X syndrome with and without autism spectrum disorder (ASD) with those of boys with Down syndrome and boys who are typically developing. METHOD: Participants were boys who have fragile X syndrome with (n = 26) and without (n = 28) ASD, boys with Down syndrome (n = 29), and boys who are typically developing (n = 22). Turns during an examiner-child interaction consisting of structured and semistructured activities were coded for the boys' ability to maintain a topic of conversation and the frequency of perseveration. RESULTS: The results revealed that boys who had both fragile X and ASD produced significantly more noncontingent discourse than did boys who had only fragile X, boys with Down syndrome, or typically developing boys. This finding was observed regardless of whether the topic was maintained or changed and whether the turn type was a response or initiation. Regardless of autism status, boys with fragile X used more perseveration than did boys in the other groups. CONCLUSION: These findings suggest that some aspects of the conversational discourse difficulties attributed to fragile X syndrome may be a function of the high rate of comorbidity between fragile X and autism, whereas some difficulties may be characteristic of fragile X syndrome. PMID- 17463243 TI - Assessing cortisol reactivity to a linguistic task as a marker of stress in individuals with left-hemisphere stroke and aphasia. AB - PURPOSE: In this study, the authors explore a method of measuring physiologic and perceived stress in individuals with aphasia by investigating salivary cortisol reactivity and subjectively perceived stress in response to a standardized linguistic task. METHOD: Fifteen individuals with aphasia and 15 age-matched healthy controls participated in a linguistic task involving speaking to an unfamiliar listener and a nonlinguistic task consisting of the Mirror Drawing Test (Starch, 1910). Salivary cortisol samples were taken following a 30-min baseline period, at the beginning and end of each task, and at 10-min intervals throughout the posttask period. Perceptions of stress also were assessed. RESULTS: No significant difference was found in cortisol levels over time within the group with aphasia between the linguistic and nonlinguistic task; however, the control group demonstrated greater cortisol reactivity during the linguistic task than during the nonlinguistic task. For the linguistic task only, the control group demonstrated greater cortisol reactivity than did the group with aphasia. Both groups perceived greater stress posttask than pretask, although the aphasia group perceived greater stress than did the control group. CONCLUSION: Adults with aphasia perceived greater stress than did healthy controls; however, this paradigm did not stimulate salivary cortisol reactivity in the aphasia group. A potential reason for this may be differences in the type or degree of psychosocial variables that are important in modulating stress in this population. Other considerations when developing methods for assessing physiologic stress include habituation and hypothalamic-pituitary-adrenal (HPA) axis dysregulation related to the neurological changes poststroke. PMID- 17463244 TI - Longitudinal relationships between lexical and grammatical development in typical and late-talking children. AB - PURPOSE: This study examined the longitudinal relationships between lexical and grammatical development in typically developing (TD) and late-talking children for the purposes of testing the single-mechanism account of language acquisition and comparing the developmental trajectories of lexical and grammatical development in late-talking and TD children. METHOD: Participants included 30 children identified as late talkers (LTs) at 2;0 (years;months), and 30 TD children matched on age, nonverbal cognition, socioeconomic status, and gender. Data were collected at 5 points between 2;0 and 5;6. RESULTS: Cross-lagged correlational analyses indicated that TD children showed evidence of bidirectional bootstrapping between lexical and grammatical development between 2;0 and 3;6. Compared with the TD group, LTs exhibited less evidence of syntactic bootstrapping. Linear mixed-effects modeling of language sample data suggested that the relationship between lexical and grammatical growth was similar for the 2 groups. CONCLUSION: Lexical and grammatical development were strongly related in both groups, consistent with the single-mechanism account of language acquisition. The results were mixed in terms of finding longitudinal differences in lexical-grammatical relationships between the TD and late-talking children; however, several analyses suggested that for late-talking children, syntactic growth may be less facilitative of lexical development. PMID- 17463245 TI - Predicting language outcomes for internationally adopted children. AB - PURPOSE: Language and speech are difficult to assess in newly arrived internationally adopted children. The purpose of this study was to determine if assessments completed when toddlers were first adopted could predict language outcomes at age 2. Local norms were used to develop early intervention guidelines that were evaluated against age 2 outcomes. Patterns of language emergence were also analyzed. METHOD: Twenty-seven children between 11 and 23 months of age adopted from Eastern Europe were followed from adoption through the 1st year home. Results from initial assessments using the Communication and Symbolic Behavior Scales-Developmental Profile (CSBS-DP; A. Wetherby & B. Prizant, 2002) and MacArthur Communicative Development Inventory-Words and Gestures (MCDI-WG; L. Fenson et al., 1993) were compared against speech and language outcomes 1 year later when the children were 2 years of age. RESULTS: By age 2, receptive language and articulation were developing well; expressive language was still emerging. Initial assessment using the CSBS-DP Behavior Sample and MCDI-WG Words Understood Developmental Quotient predicted age 2 language outcomes. Early intervention guidelines based on these 2 measures had strong positive and negative likelihood ratios (LR) when using age 2 outcomes as the criteria (LR+ = 21.00; LR- = .00). Six of the 27 children (22%) had slow language development in comparison to their peers. CONCLUSION: Newly adopted children with delays on prelinguistic and vocabulary comprehension measures were highly likely to have slow language development at age 2. Initial assessments of these abilities should be used to make early intervention decisions. PMID- 17463246 TI - Genome-wide association analysis identifies loci for type 2 diabetes and triglyceride levels. AB - New strategies for prevention and treatment of type 2 diabetes (T2D) require improved insight into disease etiology. We analyzed 386,731 common single nucleotide polymorphisms (SNPs) in 1464 patients with T2D and 1467 matched controls, each characterized for measures of glucose metabolism, lipids, obesity, and blood pressure. With collaborators (FUSION and WTCCC/UKT2D), we identified and confirmed three loci associated with T2D-in a noncoding region near CDKN2A and CDKN2B, in an intron of IGF2BP2, and an intron of CDKAL1-and replicated associations near HHEX and in SLC30A8 found by a recent whole-genome association study. We identified and confirmed association of a SNP in an intron of glucokinase regulatory protein (GCKR) with serum triglycerides. The discovery of associated variants in unsuspected genes and outside coding regions illustrates the ability of genome-wide association studies to provide potentially important clues to the pathogenesis of common diseases. PMID- 17463247 TI - Revisiting the role of the mother centriole in centriole biogenesis. AB - Centrioles duplicate once in each cell division cycle through so-called templated or canonical duplication. SAK, also called PLK4 (SAK/PLK4), a kinase implicated in tumor development, is an upstream regulator of canonical biogenesis necessary for centriole formation. We found that overexpression of SAK/PLK4 could induce amplification of centrioles in Drosophila embryos and their de novo formation in unfertilized eggs. Both processes required the activity of DSAS-6 and DSAS-4, two molecules required for canonical duplication. Thus, centriole biogenesis is a template-free self-assembly process triggered and regulated by molecules that ordinarily associate with the existing centriole. The mother centriole is not a bona fide template but a platform for a set of regulatory molecules that catalyzes and regulates daughter centriole assembly. PMID- 17463248 TI - A genome-wide association study of type 2 diabetes in Finns detects multiple susceptibility variants. AB - Identifying the genetic variants that increase the risk of type 2 diabetes (T2D) in humans has been a formidable challenge. Adopting a genome-wide association strategy, we genotyped 1161 Finnish T2D cases and 1174 Finnish normal glucose tolerant (NGT) controls with >315,000 single-nucleotide polymorphisms (SNPs) and imputed genotypes for an additional >2 million autosomal SNPs. We carried out association analysis with these SNPs to identify genetic variants that predispose to T2D, compared our T2D association results with the results of two similar studies, and genotyped 80 SNPs in an additional 1215 Finnish T2D cases and 1258 Finnish NGT controls. We identify T2D-associated variants in an intergenic region of chromosome 11p12, contribute to the identification of T2D-associated variants near the genes IGF2BP2 and CDKAL1 and the region of CDKN2A and CDKN2B, and confirm that variants near TCF7L2, SLC30A8, HHEX, FTO, PPARG, and KCNJ11 are associated with T2D risk. This brings the number of T2D loci now confidently identified to at least 10. PMID- 17463249 TI - Replication of genome-wide association signals in UK samples reveals risk loci for type 2 diabetes. AB - The molecular mechanisms involved in the development of type 2 diabetes are poorly understood. Starting from genome-wide genotype data for 1924 diabetic cases and 2938 population controls generated by the Wellcome Trust Case Control Consortium, we set out to detect replicated diabetes association signals through analysis of 3757 additional cases and 5346 controls and by integration of our findings with equivalent data from other international consortia. We detected diabetes susceptibility loci in and around the genes CDKAL1, CDKN2A/CDKN2B, and IGF2BP2 and confirmed the recently described associations at HHEX/IDE and SLC30A8. Our findings provide insight into the genetic architecture of type 2 diabetes, emphasizing the contribution of multiple variants of modest effect. The regions identified underscore the importance of pathways influencing pancreatic beta cell development and function in the etiology of type 2 diabetes. PMID- 17463250 TI - MET amplification leads to gefitinib resistance in lung cancer by activating ERBB3 signaling. AB - The epidermal growth factor receptor (EGFR) kinase inhibitors gefitinib and erlotinib are effective treatments for lung cancers with EGFR activating mutations, but these tumors invariably develop drug resistance. Here, we describe a gefitinib-sensitive lung cancer cell line that developed resistance to gefitinib as a result of focal amplification of the MET proto-oncogene. inhibition of MET signaling in these cells restored their sensitivity to gefitinib. MET amplification was detected in 4 of 18 (22%) lung cancer specimens that had developed resistance to gefitinib or erlotinib. We find that amplification of MET causes gefitinib resistance by driving ERBB3 (HER3) dependent activation of PI3K, a pathway thought to be specific to EGFR/ERBB family receptors. Thus, we propose that MET amplification may promote drug resistance in other ERBB-driven cancers as well. PMID- 17463251 TI - SCFFbxl3 controls the oscillation of the circadian clock by directing the degradation of cryptochrome proteins. AB - One component of the circadian clock in mammals is the Clock-Bmal1 heterodimeric transcription factor. Among its downstream targets, two genes, Cry1 and Cry2, encode inhibitors of the Clock-Bmal1 complex that establish a negative-feedback loop. We found that both Cry1 and Cry2 proteins are ubiquitinated and degraded via the SCF(Fbxl3) ubiquitin ligase complex. This regulation by SCF(Fbxl3) is a prerequisite for the efficient and timely reactivation of Clock-Bmal1 and the consequent expression of Per1 and Per2, two regulators of the circadian clock that display tumor suppressor activity. Silencing of Fbxl3 produced no effect in Cry1-/-;Cry2-/- cells, which shows that Fbxl3 controls clock oscillations by mediating the degradation of CRY proteins. PMID- 17463252 TI - The after-hours mutant reveals a role for Fbxl3 in determining mammalian circadian period. AB - By screening N-ethyl-N-nitrosourea-mutagenized animals for alterations in rhythms of wheel-running activity, we identified a mouse mutation, after hours (Afh). The mutation, a Cys(358)Ser substitution in Fbxl3, an F-box protein with leucine-rich repeats, results in long free-running rhythms of about 27 hours in homozygotes. Circadian transcriptional and translational oscillations are attenuated in Afh mice. The Afh allele significantly affected Per2 expression and delayed the rate of Cry protein degradation in Per2::Luciferase tissue slices. Our in vivo and in vitro studies reveal a central role for Fbxl3 in mammalian circadian timekeeping. PMID- 17463253 TI - The biofuels conundrum. PMID- 17463254 TI - Stem cells. Stem cell president quits after acrimonious meeting. PMID- 17463255 TI - Geochemistry. Humongous eruptions linked to dramatic environmental changes. PMID- 17463256 TI - Lunar science. Congress restores funds for NASA robotic landers. PMID- 17463257 TI - Exoplanets. Habitable, but not much like home. PMID- 17463258 TI - Biodefense. Proposed biosecurity review plan endorses self-regulation. PMID- 17463259 TI - Genetics. Erasing microRNAs reveals their powerful punch. PMID- 17463260 TI - Big facilities. Researchers get in synch down under. PMID- 17463261 TI - Marine biology. Killing whales for science? PMID- 17463262 TI - Cross-cultural research. Pentagon asks academics for help in understanding its enemies. PMID- 17463263 TI - Carbon emissions. Improved monitoring of rainforests helps pierce haze of deforestation. PMID- 17463264 TI - American Physical Society meeting. Gravity probe researchers report 'glimpses' of long-awaited payoff. PMID- 17463265 TI - American Physical Society meeting. Neutrino study finds four's a crowd. PMID- 17463266 TI - American Physical Society meeting. Snapshots from the meeting. PMID- 17463267 TI - Health clues from polar regions. PMID- 17463268 TI - Science, religion, and climate change. PMID- 17463269 TI - Clarifying a quote on women in science. PMID- 17463270 TI - Notes on modeling light water reactors. PMID- 17463271 TI - The evolution of eukaryotes. PMID- 17463272 TI - Environment. Building a "green" railway in China. PMID- 17463273 TI - The pipeline. Benefits of undergraduate research experiences. PMID- 17463274 TI - Genetics. Getting closer to the whole picture. PMID- 17463275 TI - Neuroscience. How to fill a synapse. PMID- 17463276 TI - Chemistry. A promising mimic of hydrogenase activity. PMID- 17463277 TI - Physics. Factoring numbers with waves. PMID- 17463278 TI - Physics. The end of an entanglement. PMID- 17463279 TI - Evolution. Oxygen and evolution. PMID- 17463280 TI - The problem with determining atomic structure at the nanoscale. AB - Emerging complex functional materials often have atomic order limited to the nanoscale. Examples include nanoparticles, species encapsulated in mesoporous hosts, and bulk crystals with intrinsic nanoscale order. The powerful methods that we have for solving the atomic structure of bulk crystals fail for such materials. Currently, no broadly applicable, quantitative, and robust methods exist to replace crystallography at the nanoscale. We provide an overview of various classes of nanostructured materials and review the methods that are currently used to study their structure. We suggest that successful solutions to these nanostructure problems will involve interactions among researchers from materials science, physics, chemistry, computer science, and applied mathematics, working within a "complex modeling" paradigm that combines theory and experiment in a self-consistent computational framework. PMID- 17463281 TI - Fast routing in road networks with transit nodes. AB - When you drive to somewhere far away, you will leave your current location via one of only a few important traffic junctions. Starting from this informal observation, we developed an algorithmic approach, transit node routing, that allows us to reduce quickest path queries in road networks to a small number of table lookups. For road maps of Western Europe and the United States, our best query times improved over the best previously published figures by two orders of magnitude. This is also more than one million times faster than the best known algorithm for general networks. PMID- 17463282 TI - Revisiting carbon flux through the ocean's twilight zone. AB - The oceanic biological pump drives sequestration of carbon dioxide in the deep sea via sinking particles. Rapid biological consumption and remineralization of carbon in the "twilight zone" (depths between the euphotic zone and 1000 meters) reduce the efficiency of sequestration. By using neutrally buoyant sediment traps to sample this chronically understudied realm, we measured a transfer efficiency of sinking particulate organic carbon between 150 and 500 meters of 20 and 50% at two contrasting sites. This large variability in transfer efficiency is poorly represented in biogeochemical models. If applied globally, this is equivalent to a difference in carbon sequestration of more than 3 petagrams of carbon per year. PMID- 17463283 TI - A selective activity-dependent requirement for dynamin 1 in synaptic vesicle endocytosis. AB - Dynamin 1 is a neuron-specific guanosine triphosphatase thought to be critically required for the fission reaction of synaptic vesicle endocytosis. Unexpectedly, mice lacking dynamin 1 were able to form functional synapses, even though their postnatal viability was limited. However, during spontaneous network activity, branched, tubular plasma membrane invaginations accumulated, capped by clathrin coated pits, in synapses of dynamin 1-knockout mice. Synaptic vesicle endocytosis was severely impaired during strong exogenous stimulation but resumed efficiently when the stimulus was terminated. Thus, dynamin 1-independent mechanisms can support limited synaptic vesicle endocytosis, but dynamin 1 is needed during high levels of neuronal activity. PMID- 17463284 TI - Environment-induced sudden death of entanglement. AB - We demonstrate the difference between local, single-particle dynamics and global dynamics of entangled quantum systems coupled to independent environments. Using an all-optical experimental setup, we showed that, even when the environment induced decay of each system is asymptotic, quantum entanglement may suddenly disappear. This "sudden death" constitutes yet another distinct and counterintuitive trait of entanglement. PMID- 17463285 TI - A dinuclear Ni(mu-H)Ru complex derived from H2. AB - Models of the active site in [NiFe]hydrogenase enzymes have proven challenging to prepare. We isolated a paramagnetic dinuclear nickel-ruthenium complex with a bridging hydrido ligand from the heterolytic cleavage of H2 by a dinuclear NiRu aqua complex in water under ambient conditions (20 degrees C and 1 atmosphere pressure). The structure of the hexacoordinate Ni(mu-H)Ru complex was unequivocally determined by neutron diffraction analysis, and it comes closest to an effective analog for the core structure of the proposed active form of the enzyme. PMID- 17463286 TI - Paleocene-Eocene thermal maximum and the opening of the Northeast Atlantic. AB - The Paleocene-Eocene thermal maximum (PETM) has been attributed to a sudden release of carbon dioxide and/or methane. 40Ar/39Ar age determinations show that the Danish Ash-17 deposit, which overlies the PETM by about 450,000 years in the Atlantic, and the Skraenterne Formation Tuff, representing the end of 1 +/- 0.5 million years of massive volcanism in East Greenland, are coeval. The relative age of Danish Ash-17 thus places the PETM onset after the beginning of massive flood basalt volcanism at 56.1 +/- 0.4 million years ago but within error of the estimated continental breakup time of 55.5 +/- 0.3 million years ago, marked by the eruption of mid-ocean ridge basalt-like flows. These correlations support the view that the PETM was triggered by greenhouse gas release during magma interaction with basin-filling carbon-rich sedimentary rocks proximal to the embryonic plate boundary between Greenland and Europe. PMID- 17463287 TI - NOV (CCN3) functions as a regulator of human hematopoietic stem or progenitor cells. AB - Clinically successful hematopoietic cell transplantation is dependent on hematopoietic stem and progenitor cells. Here we identify the matricellular protein Nephroblastoma Overexpressed (Nov, CCN3) as being essential for their functional integrity. Nov expression is restricted to the primitive (CD34) compartments of umbilical vein cord blood, and its knockdown in these cells by lentivirus-mediated RNA interference abrogates their function in vitro and in vivo. Conversely, forced expression of Nov and addition of recombinant Nov protein both enhance primitive stem and/or progenitor activity. Taken together, our results identify Nov (CCN3) as a regulator of human hematopoietic stem or progenitor cells. PMID- 17463288 TI - Modeling the initiation and progression of human acute leukemia in mice. AB - Our understanding of leukemia development and progression has been hampered by the lack of in vivo models in which disease is initiated from primary human hematopoietic cells. We showed that upon transplantation into immunodeficient mice, primitive human hematopoietic cells expressing a mixed-lineage leukemia (MLL) fusion gene generated myeloid or lymphoid acute leukemias, with features that recapitulated human diseases. Analysis of serially transplanted mice revealed that the disease is sustained by leukemia-initiating cells (L-ICs) that have evolved over time from a primitive cell type with a germline immunoglobulin heavy chain (IgH) gene configuration to a cell type containing rearranged IgH genes. The L-ICs retained both myeloid and lymphoid lineage potential and remained responsive to microenvironmental cues. The properties of these cells provide a biological basis for several clinical hallmarks of MLL leukemias. PMID- 17463289 TI - Regulation of the germinal center response by microRNA-155. AB - MicroRNAs are small RNA species involved in biological control at multiple levels. Using genetic deletion and transgenic approaches, we show that the evolutionarily conserved microRNA-155 (miR-155) has an important role in the mammalian immune system, specifically in regulating T helper cell differentiation and the germinal center reaction to produce an optimal T cell-dependent antibody response. miR-155 exerts this control, at least in part, by regulating cytokine production. These results also suggest that individual microRNAs can exert critical control over mammalian differentiation processes in vivo. PMID- 17463290 TI - Requirement of bic/microRNA-155 for normal immune function. AB - MicroRNAs are a class of small RNAs that are increasingly being recognized as important regulators of gene expression. Although hundreds of microRNAs are present in the mammalian genome, genetic studies addressing their physiological roles are at an early stage. We have shown that mice deficient for bic/microRNA 155 are immunodeficient and display increased lung airway remodeling. We demonstrate a requirement of bic/microRNA-155 for the function of B and T lymphocytes and dendritic cells. Transcriptome analysis of bic/microRNA-155 deficient CD4+ T cells identified a wide spectrum of microRNA-155-regulated genes, including cytokines, chemokines, and transcription factors. Our work suggests that bic/microRNA-155 plays a key role in the homeostasis and function of the immune system. PMID- 17463291 TI - Distinct pathways of antigen uptake and intracellular routing in CD4 and CD8 T cell activation. AB - The mechanisms that allow antigen-presenting cells (APCs) to selectively present extracellular antigen to CD8+ effector T cells (cross-presentation) or to CD4+ T helper cells are not fully resolved. We demonstrated that APCs use distinct endocytosis mechanisms to simultaneously introduce soluble antigen into separate intracellular compartments, which were dedicated to presentation to CD8+ or CD4+ T cells. Specifically, the mannose receptor supplied an early endosomal compartment distinct from lysosomes, which was committed to cross-presentation. These findings imply that antigen does not require intracellular diversion to access the cross-presentation pathway, because it can enter the pathway already during endocytosis. PMID- 17463292 TI - PCR-based strategy to detect and identify species of Phaeoacremonium causing grapevine diseases. AB - Species of Phaeoacremonium (especially Phaeoacremonium aleophilum) are associated with two severe diseases in grapevines, Petri disease in young plants and Esca disease in adult plants. Phaeoacremonium species grow slowly on culture medium, and it is difficult to identify these species on the basis of morphological characteristics. Primers Pm1 and Pm2 were designed in the ribosomal DNA internal transcribed spacer (ITS) regions ITS1 and ITS2, respectively. They yielded a single amplicon of 415 bp for nine species of Phaeoacremonium that may occur in grapevines. A nested PCR (using general fungal primers ITS1F/ITS4 in the primary reaction) was developed to detect Phaeoacremonium directly in grapevine wood. Molecular detection was more sensitive than the traditional method of culturing in growth medium was. Identification of Phaeoacremonium species was achieved by digesting the PCR-amplified fragment with the restriction enzymes BssKI, EcoO109I, and HhaI. It was possible to distinguish these species by their restriction fragment length polymorphism patterns, except for Phaeoacremonium viticola and Phaeoacremonium angustius, which had 100% similarity in their ITS region sequences. A species-specific PCR amplification of the partial beta tubulin gene using the primer pair Pbr4_1/T1 and Pbr8/T1 was necessary to differentiate P. angustius from P. viticola, respectively. An easy and fast protocol was developed to detect and identify species of Phaeoacremonium in a few hours. Primers defined here can be used in a plant nursery sanitation program to produce plants free of Phaeoacremonium spp. Use of healthy grapevine plants in new plantations is the most effective measure to manage Petri disease. PMID- 17463293 TI - Ischemic preconditioning does not protect via blockade of electron transport. AB - Ischemic preconditioning (IPC) before sustained ischemia decreases myocardial infarct size mediated in part via protection of cardiac mitochondria. Reversible blockade of electron transport at complex I immediately before sustained ischemia also preserves mitochondrial respiration and decreases infarct size. We proposed that IPC would attenuate electron transport from complex I as a potential effector mechanism of cardioprotection. Isolated, Langendorff-perfused rat hearts underwent IPC (3 cycles of 5-min 37 degrees C global ischemia and 5-min reperfusion) or were perfused for 40 min without ischemia as controls. Subsarcolemmal (SSM) and interfibrillar (IFM) populations of mitochondria were isolated. IPC did not decrease ADP-stimulated respiration measured in intact mitochondria using substrates that donate reducing equivalents to complex I. Maximally expressed complex I activity measured as rotenone-sensitive NADH:ubiquinone oxidoreductase in detergent-solubilized mitochondria was also unaffected by IPC. Thus the protection of IPC does not occur as a consequence of a partial decrease in complex I activity leading to a decrease in integrated respiration through complex I. IPC and blockade of electron transport both converge on mitochondria as effectors of cardioprotection; however, each modulates mitochondrial metabolism during ischemia by different mechanisms to achieve cardioprotection. PMID- 17463294 TI - Thermic effect of food and beta-adrenergic thermogenic responsiveness in habitually exercising and sedentary healthy adult humans. AB - The thermic effect of food (TEF) is an important physiological determinant of total daily energy expenditure (EE) and energy balance. TEF is believed to be mediated in part by sympathetic nervous system activation and consequent beta adrenergic receptor (beta-AR) stimulation of metabolism. TEF is greater in habitually exercising than in sedentary adults, despite similar postprandial sympathetic nervous system activation. We determined whether augmented TEF in habitually exercising adults is associated with enhanced peripheral thermogenic responsiveness to beta-AR stimulation. In separate experiments in 22 sedentary and 29 habitually exercising adults, we measured the increase in EE (indirect calorimetry, ventilated hood) during beta-AR stimulation (intravenous isoproterenol: 6, 12, and 24 ng x kg fat-free mass(-1) x min(-1)) and EE before and after a liquid meal (40% of resting EE; 53% carbohydrate, 32% fat, 15% protein). The increase in EE during incremental isoproterenol administration was greater (P = 0.01) in habitual exercisers (0.34 +/- 0.03, 0.54 +/- 0.04, 0.81 +/- 0.05 kJ/min; means +/- SE) than in sedentary adults (0.26 +/- 0.03, 0.40 +/- 0.03, 0.64 +/- 0.04 kJ/min). The area under the TEF response curve was also greater (P = 0.04) in habitual exercisers (160 +/- 9 kJ) than in sedentary adults (130 +/- 11 kJ) and was positively related to beta-AR thermogenic responsiveness (r = 0.32, P = 0.02). We conclude that TEF is related to beta-AR thermogenic responsiveness and that the greater TEF in habitual exercisers is attributable in part to their augmented beta-AR thermogenic responsiveness. Our results also suggest that peripheral thermogenic responsiveness to beta-AR stimulation is a physiological determinant of TEF and hence energy balance in healthy adult humans. PMID- 17463295 TI - Role of the altitude level on cerebral autoregulation in residents at high altitude. AB - Cerebral autoregulation is impaired in Himalayan high-altitude residents who live above 4,200 m. This study was undertaken to determine the altitude at which this impairment of autoregulation occurs. A second aim of the study was to test the hypothesis that administration of oxygen can reverse this impairment in autoregulation at high altitudes. In four groups of 10 Himalayan high-altitude dwellers residing at 1,330, 2,650, 3,440, and 4,243 m, arterial oxygen saturation (Sa(O(2))), blood pressure, and middle cerebral artery blood velocity were monitored during infusion of phenylephrine to determine static cerebral autoregulation. On the basis of these measurements, the cerebral autoregulation index (AI) was calculated. Normally, AI is between zero and 1. AI of 0 implies absent autoregulation, and AI of 1 implies intact autoregulation. At 1,330 m (Sa(O(2)) = 97%), 2,650 m (Sa(O(2)) = 96%), and 3,440 m (Sa(O(2)) = 93%), AI values (mean +/- SD) were, respectively, 0.63 +/- 0.27, 0.57 +/- 0.22, and 0.57 +/- 0.15. At 4,243 m (Sa(O(2)) = 88%), AI was 0.22 +/- 0.18 (P < 0.0005, compared with AI at the lower altitudes) and increased to 0.49 +/- 0.23 (P = 0.008, paired t-test) when oxygen was administered (Sa(O(2)) = 98%). In conclusion, high altitude residents living at 4,243 m have almost total loss of cerebral autoregulation, which improved during oxygen administration. Those people living at 3,440 m and lower have still functioning cerebral autoregulation. This study showed that the altitude region between 3,440 and 4,243 m, marked by Sa(O(2)) in the high-altitude dwellers of 93% and 88%, is a transitional zone, above which cerebral autoregulation becomes critically impaired. PMID- 17463296 TI - Turning on the central contribution to contractions evoked by neuromuscular electrical stimulation. AB - Neuromuscular electrical stimulation can generate contractions through peripheral and central mechanisms. Direct activation of motor axons (peripheral mechanism) recruits motor units in an unnatural order, with fatigable muscle fibers often activated early in contractions. The activation of sensory axons can produce contractions through a central mechanism, providing excitatory synaptic input to spinal neurons that recruit motor units in the natural order. Presently, we quantified the effect of stimulation frequency (10-100 Hz), duration (0.25-2 s of high-frequency bursts, or 20 s of constant-frequency stimulation), and intensity [1-5% maximal voluntary contraction (MVC) torque generated by a brief 100-Hz train] on the torque generated centrally. Electrical stimulation (1-ms pulses) was delivered over the triceps surae in eight subjects, and plantar flexion torque was recorded. Stimulation frequency, duration, and intensity all influenced the magnitude of the central contribution to torque. Central torque did not develop at frequencies < or = 20 Hz, and it was maximal at frequencies > or = 80 Hz. Increasing the duration of high-frequency stimulation increased the central contribution to torque, as central torque developed over 11 s. Central torque was greatest at a relatively low contraction intensity. The largest amount of central torque was produced by a 20-s, 100-Hz train (10.7 +/- 5.5 %MVC) and by repeated 2-s bursts of 80- or 100-Hz stimulation (9.2 +/- 4.8 and 10.2 +/- 8.1% MVC, respectively). Therefore, central torque was maximized by applying high frequency, long-duration stimulation while avoiding antidromic block by stimulating at a relatively low intensity. If, as hypothesized, the central mechanism primarily activates fatigue-resistant muscle fibers, generating muscle contractions through this pathway may improve rehabilitation applications. PMID- 17463297 TI - Preexercise sodium loading aids fluid balance and endurance for women exercising in the heat. AB - This study was conducted during the high-hormone phase of both natural and oral contraceptive pill (OCP)-mediated menstrual cycles to determine whether preexercise ingestion of a concentrated sodium beverage would increase plasma volume (PV), reduce physiological strain, and aid endurance of moderately trained women cycling in warm conditions. Thirteen trained cyclists [peak O(2) uptake 52 ml x kg(-1) x min(-1) (SD 2), age 26 yr (SD 6), weight 60.8 kg (SD 5)] who were oral contraceptive users (n = 6) or not (n = 7) completed this double-blind, crossover experiment. Cyclists ingested a concentrated-sodium (High Na(+): 164 mmol Na(+)/l) or low-sodium (Low Na(+): 10 mmol Na(+)/l) beverage (10 ml/kg) before cycling to exhaustion at 70% Peak O(2) uptake in warm conditions (32 degrees C, 50% relative humidity, air velocity 4.5 m/s). Beverage (approximately 628 ml) was ingested in seven portions across 60 min beginning 105 min before exercise, with no additional fluid given until the end of the trial. Trials were separated by one to two menstrual cycles. High Na(+) increased PV (calculated from hematocrit and hemoglobin concentration) before exercise, whereas Low Na(+) did not [-4.4 (SD 1.1) vs. -1.9% (SD 1.3); 95% confidence interval: for the difference 5.20, 6.92; P < 0.0001], and it involved greater time to exhaustion [98.8 (SD 25.6) vs. 78.7 (SD 24.6) min; 95% confidence interval: 13.3, 26.8; P < 0.0001]. Core temperature rose more quickly with Low Na(+) [1.6 degrees C/h (SD 0.2)] than High Na(+) [1.2 degrees C/h (SD 0.2); P = 0.04]. Plasma [AVP], [Na(+)] concentration, and osmolality, and urine volume, [Na(+)], and osmolality decreased with sodium loading (P < 0.05) independent of pill usage. Thus preexercise ingestion of a concentrated sodium beverage increased PV, reduced thermoregulatory strain, and increased exercise capacity for women in the high hormone phase of natural and oral contraceptive pill-mediated menstrual cycles, in warm conditions. PMID- 17463298 TI - Hemodynamic consequences of rapid changes in posture in humans. AB - Tolerance to +G(z) gravitational stress is reduced when +G(z) stress is preceded by exposure to hypogravity (fraction, 0, or negative G(z)). For example, there is an exaggerated fall in eye-level arterial pressure (ELAP) early on during +G(z) stress (head-up tilt; HUT) when this stress is immediately preceded by -G(z) stress (head-down tilt; HDT). The aims of the present study were to characterize the hemodynamic consequences of brief HDT on subsequent HUT and to test the hypothesis that an elevation in leg vascular conductance induced by -G(z) stress contributes to the exaggerated fall in ELAP. Young healthy subjects (n = 3 men and 4 women) were subjected to 30 s of 30 degrees HUT from a horizontal position and to 30 s of 30 degrees HUT when HUT was immediately preceded by 20 s of -15 degrees HDT. Four bouts of HDT-HUT were alternated between five bouts of HUT in a counterbalanced designed to minimize possible time effects of repeated exposure to gravitational stress. One minute was allowed for recovery between tilts. Brief exposure to HDT elicited an exaggerated fall in ELAP during the first seconds of the subsequent HUT (-17.9 +/- 1.4 mmHg) compared with HUT alone (-12.4 +/- 1.2 mmHg, P <0.05) despite a greater rise in stroke volume (Doppler ultrasound) and cardiac output over this brief time period in the HDT-HUT trials compared with the HUT trials (thereafter stroke volume fell under both conditions). The greater fall in ELAP was associated with an exaggerated increase in leg blood flow (femoral artery Doppler ultrasound) and was therefore largely (70%) attributable to an exaggerated rise in estimated leg vascular conductance, confirming our hypotheses. Thus brief exposure to -G(z) stress leads to an exaggerated fall in ELAP during subsequent HUT, owing to an exaggerated increase in estimated leg vascular conductance. PMID- 17463300 TI - Influence of noninvasive peripheral arterial blood pressure measurements on assessment of dynamic cerebral autoregulation. AB - Assessment of dynamic cerebral autoregulation (CA) requires continuous recording of arterial blood pressure (ABP). In humans, noninvasive ABP recordings with the Finapres device have often been used for this purpose. We compared estimates of dynamic CA derived from Finapres with those from invasive recordings in the aorta. Measurements of finger noninvasive ABP (Finapres), intra-aortic ABP (Millar catheter), surface ECG, transcutaneous CO2, and bilateral cerebral blood flow velocity (CBFV) in the middle cerebral arteries were simultaneously and continuously recorded in 27 patients scheduled for percutaneous coronary interventions. Phase, gain, coherence, and CBFV step response from both the Finapres and intra-arterial catheter were estimated by transfer function analysis. A dynamic autoregulation index (ARI) was also calculated. For both hemispheres, the ARI index and the CBFV step response recovery at 4 s were significantly greater for the Finapres-derived estimates than for the values obtained from aortic pressure. The transfer function gain for frequencies <0.1 Hz was significantly smaller for the Finapres estimates. The phase frequency response was significantly greater for the Finapres estimates at frequencies >0.1 Hz, but not at lower frequencies. The Finapres gives higher values for the efficiency of dynamic CA compared with values derived from aortic pressure measurements, as indicated by biases in the ARI index, CBFV step response, gain, and phase. Despite the significance of these biases, their relatively small amplitude indicates a good level of agreement between indexes of CA derived from the Finapres compared with corresponding estimates obtained from invasive measurements of aortic ABP. PMID- 17463301 TI - Chronic intermittent hypoxia modulates eosinophil- and neutrophil-platelet aggregation and inflammatory cytokine secretion caused by strenuous exercise in men. AB - Although acclimatization to intermittent hypoxia (IH) improves exercise performance by increasing oxygen delivery and utilization, the effects of chronic IH on platelet-leukocyte interaction and inflammation-related cytokine secretion caused by strenuous exercise remain unclear. This investigation elucidates how two intensities of IH influence eosinophil- and neutrophil-platelet aggregation (EPA and NPA) as well as pro- and anti-inflammatory cytokines mediated by strenuous exercise. Twenty healthy sedentary men were randomly divided into severe (SIH) and moderate (MIH) IH groups; groups were exposed to 12% O2 (SIH) and 15% O2 (MIH) for 1 h/day, respectively, for 5 days/wk for 8 wk in a normobaric hypoxia chamber. Before IH intervention, 1) exercise up to maximal oxygen consumption promoted shear stress-, LPS-, and N-formyl-methionyl-leucyl phenylalanine-induced EPA, increased IL-1beta and malondialdehyde levels, and decreased total antioxidant levels in plasma and 2) exposure to 12% O2, but not to 15% O2 for 1 h, enhanced LPS-induced EPA and reduced plasma total antioxidant levels. After IH for 8 wk, hypoxia- and exercise-promoted EPA, IL-1beta, or malondialdehyde levels were suppressed in both MIH and SIH groups, and plasma IL 6 and IL-10 levels in the SIH group were increased. However, the NPA induced by the shear force and chemical agonists was not changed under the two IH regimens. Therefore, both MIH and SIH regimens ameliorate eosinophil- and platelet-related thrombosis, proinflammatory IL-1beta secretion, and lipid peroxidation enhanced by strenuous exercise. Furthermore, SIH simultaneously increases circulatory anti inflammatory IL-6 and IL-10 concentrations. These findings can help to develop effective IH regimens that improve aerobic fitness and minimize risk of thromboinflammation. PMID- 17463302 TI - Sustained contraction at very low forces produces prominent supraspinal fatigue in human elbow flexor muscles. AB - During sustained maximal voluntary contractions (MVCs), most fatigue occurs within the muscle, but some occurs because voluntary activation of the muscle declines (central fatigue), and some of this reflects suboptimal output from the motor cortex (supraspinal fatigue). This study examines whether supraspinal fatigue occurs during a sustained submaximal contraction of 5% MVC. Eight subjects sustained an isometric elbow flexion of 5% MVC for 70 min. Brief MVCs were performed every 3 min, with stimulation of the motor point, motor cortex, and brachial plexus. Perceived effort and pain, elbow flexion torque, and surface EMGs from biceps and brachioradialis were recorded. During the sustained 5% contraction, perceived effort increased from 0.5 to 3.9 (out of 10), and elbow flexor EMG increased steadily by approximately 60-80%. Torque during brief MVCs fell to 72% of control values, while both the resting twitch and EMG declined progressively. Thus the sustained weak contraction caused fatigue, some of which was due to peripheral mechanisms. Voluntary activation measured by motor point and motor cortex stimulation methods fell to 90% and 80%, respectively. Thus some of the fatigue was central. Calculations based on the fall in voluntary activation measured with cortical stimulation indicate that about two-thirds of the fatigue was due to supraspinal mechanisms. Therefore, sustained performance of a very low-force contraction produces a progressive inability to drive the motor cortex optimally during brief MVCs. The effect of central fatigue on performance of the weak contraction is less clear, but it may contribute to the increase in perceived effort. PMID- 17463303 TI - AMP deaminase deficiency is associated with lower sprint cycling performance in healthy subjects. AB - AMP deaminase (AMPD) deficiency is an inherited disorder of skeletal muscle found in approximately 2% of the Caucasian population. Although most AMPD-deficient individuals are asymptomatic, a small subset has exercise-related cramping and pain without any other identifiable neuromuscular complications. This heterogeneity has raised doubts about the physiological significance of AMPD in skeletal muscle, despite evidence for disrupted adenine nucleotide catabolism during exercise in deficient individuals. Previous studies have evaluated the effect of AMPD deficiency on exercise performance with mixed results. This study was designed to circumvent the perceived limitations in previous reports by measuring exercise performance during a 30-s Wingate test in 139 healthy, physically active subjects of both sexes, with different AMPD1 genotypes, including 12 AMPD-deficient subjects. Three of the deficient subjects were compound heterozygotes characterized by the common c.34C>T mutation in one allele and a newly discovered AMPD1 mutation, c.404delT, in the other. While there was no significant difference in peak power across AMPD1 genotypes, statistical analysis revealed a faster power decrease in the AMPD-deficient group and a difference in mean power across the genotypes (P = 0.0035). This divergence was most striking at 15 s of the 30-s cycling. Assessed by the fatigue index, the decrease in power output at 15 s of exercise was significantly greater in the deficient group compared with the other genotypes (P = 0.0006). The approximate 10% lower mean power in healthy AMPD-deficient subjects during a 30-s Wingate cycling test reveals a functional role for the AMPD1 enzyme in sprint exercise. PMID- 17463304 TI - The influence of anti-inflammatory medication on exercise-induced myogenic precursor cell responses in humans. AB - The consumption of nonsteroidal anti-inflammatory drugs (NSAIDs) is widespread among athletes when faced with muscle soreness or injury, but the effects of NSAIDs on satellite cell activity in humans are unknown. To investigate this, 14 healthy male endurance athletes (mean peak oxygen consumption 62 ml x kg(-1) x min(-1)) volunteered for the study, which involved running 36 km. They were divided into two groups and received either 100 mg indomethacin per day or placebo. Muscle biopsies collected before the run and on days 1, 3, and 8 afterward were analyzed for satellite cells by immunohistochemistry with the aid of neural cell adhesion molecule (NCAM) and fetal antigen-1 (FA1) antibodies. Muscle biopsies were also collected from untrained individuals for comparison. Compared with preexercise levels, a 27% increase in the number of NCAM+ cells was observed on day 8 postexercise in the placebo group (P < 0.05), while levels remained similar at all time points in the NSAID group. No change was seen in the proportion of FA1+ cells, although lower levels were found in the muscle of endurance-trained athletes compared with untrained individuals (P < 0.05). These results suggest that ingestion of anti-inflammatory drugs attenuates the exercise induced increase in satellite cell number, supporting the role of the cyclooxygenase pathway in satellite cell activity. PMID- 17463305 TI - Hierarchy of individual calibration levels for heart rate and accelerometry to measure physical activity. AB - Combining accelerometry with heart rate (HR) monitoring may improve precision of physical activity measurement. Considerable variation exists in the relationships between physical activity intensity (PAI) and HR and accelerometry, which may be reduced by individual calibration. However, individual calibration limits feasibility of these techniques in population studies, and less burdensome, yet valid, methods of calibration are required. We aimed to evaluate the precision of different individual calibration procedures against a reference calibration procedure: a ramped treadmill walking-running test with continuous measurement of PAI by indirect calorimetry in 26 women and 25 men [mean (SD): 35 (9) yr, 1.69 (0.10) m, 70 (14) kg]. Acceleration (along the longitudinal axis of the trunk) and HR were measured simultaneously. Alternative calibration procedures included treadmill testing without calorimetry, submaximal step and walk tests with and without calorimetry, and nonexercise calibration using sleeping HR and gender. Reference accelerometry and HR models explained >95% of the between-individual variance in PAI (P < 0.001). This fraction dropped to 73 and 81%, respectively, for accelerometry and HR models calibrated with treadmill tests without calorimetry. Step-test calibration captured 62-64% (accelerometry) and 68% (HR) of the variance between individuals. Corresponding values were 63-76% and 59-61% for walk-test calibration. There was only little benefit of including calorimetry during step and walk calibration for HR models. Nonexercise calibration procedures explained 54% (accelerometry) and 30% (HR) of the between-individual variance. In conclusion, a substantial proportion of the between-individual variance in relationships between PAI, accelerometry, and HR is captured with simple calibration procedures, feasible for use in epidemiological studies. PMID- 17463306 TI - Herbal medicine in stroke: does it have a future? PMID- 17463307 TI - Carotid angioplasty and stent placement after EVA-3S trial. PMID- 17463308 TI - Stroke units in their natural habitat: systematic review of observational studies. AB - BACKGROUND AND PURPOSE: Within clinical trials, stroke patients allocated to receive organized inpatient (stroke unit) care are more likely to survive, return home, and regain independence than those allocated to conventional care. However, there are concerns that the benefits seen in clinical trials may not be replicated in routine practice. We carried out a systematic review of observational studies of stroke unit implementation. METHODS: We searched (up to January 2006) MEDLINE, EMBASE, CINAHL, Cochrane Library, British Nursing Index, Cochrane Stroke Group register, and recent conference abstracts for observational studies that compared the outcomes of stroke patients managed in a stroke unit versus non-stroke unit care. We excluded studies that did not describe either matching for baseline prognostic factors or adjustment for case-mix characteristics. The primary outcome was death within 1 year. We also recorded poor outcome (death, institutional care, or dependency). Data analysis used the generic inverse variance method in Revman 4.2. Where raw data were provided, effect sizes and variances were calculated accordingly. We used a random-effects model and explored for sources of heterogeneity. RESULTS: We identified 72 articles describing stroke unit outcomes; 25 were eligible for review; and 18 provided data on case fatality or poor outcome. Stroke unit care was associated with significantly reduced odds of death (odds ratio=0.79, 95% CI=0.73 to 0.86; P<0.00001) and of death or poor outcome (odds ratio=0.87, 95% CI=0.80 to 0.95; P=0.002) within 1 year of stroke. Results were complicated by significant heterogeneity (P<0.05), mainly in single-center studies. CONCLUSIONS: Although these results are complicated by potential bias and heterogeneity, the observed benefit associated with stroke unit care in routine practice is comparable to that in clinical trials. PMID- 17463309 TI - Power M-Mode Doppler and single-gate spectral analysis using a 2-MHz pulsed-wave Doppler transducer to directly detect cervical internal carotid artery stenosis: use of the continuity principle: report of a novel technique. AB - BACKGROUND AND PURPOSE: We hypothesized that direct cervical investigation with Power M-Mode Doppler (PMD) combined with single-gate Doppler spectral analysis (SGDSA) using a 2-MHz pulsed-wave Doppler transducer would show reasonable accuracy parameters when compared with standard color-coded carotid duplex ultrasound (CDU). METHODS: We prospectively screened for cervical internal carotid artery (ICA) stenosis by direct observation using a 2 MHz PMD/SGDSA device. PMD identified the artery (location, depth, flow direction) and SGDSA assessed waveform; peak systolic, end diastolic, and mean flow velocities (MFV) of the common carotid artery; cervical ICA proximally and distally; and external carotid artery. Diagnostic accuracy was compared with concurrent carotid duplex ultrasound. The continuity principle was applied using the proximal/distal cervical ICA MFV ratio. RESULTS: We examined 456 vessels (228 patients). Using ICA proximally/ICA distally MFV ratio of 1.5 or greater or absence of ICA signature, for 40% to 59% or greater stenosis (including occlusions), sensitivity was 75.4%, specificity 99.8%, positive predictive value 97.7%, negative predictive value 96.6%, and accuracy 96.7%. For MFV ratio 1.6 or greater or absence of ICA signature and 60% to 79% or greater stenosis (including occlusions), sensitivity was 92.3%, specificity 98.1%, positive predictive value 81.8%, negative predictive value 99.3%, and accuracy 97.6%. CONCLUSIONS: Use of combined PMD and SGDSA to directly observe the extracranial ICA is reasonably accurate compared with carotid duplex ultrasound. Using the MFV ratio of proximal/distal extracranial ICA improves accuracy parameters and provides a quick and effective bedside screen for ICA stenosis. This novel technique should be considered part of the standard PMD/transcranial Doppler examination. PMID- 17463310 TI - GPx-3 gene promoter variation and the risk of arterial ischemic stroke. PMID- 17463311 TI - Reproducibility of high-resolution MRI for the identification and the quantification of carotid atherosclerotic plaque components: consequences for prognosis studies and therapeutic trials. AB - BACKGROUND AND PURPOSE: Although MRI is increasingly proposed to investigate composition of carotid atherosclerosis, its reproducibility has rarely been addressed. We assessed the reproducibility of MRI for the identification and quantification of carotid atherosclerotic plaque components. METHODS: Using published criteria, 2 readers independently analyzed the carotid MRI (1.5-T MR units with a 4-channel phased-array surface coil, Machnet) of 85 consecutive patients with symptomatic (40% to 69% according to NASCET method) or asymptomatic (60% or greater) carotid artery stenosis enrolled in an ongoing prognostic study. One reader reevaluated all images. Fibrous cap was also secondarily identified independently on T2-weighted and time-of-flight (TOF) images. RESULTS: Intraobserver agreement was substantial for the identification of calcifications (kappa [kappa]=0.70; 95% CI: 0.54 to 0.86) and lipid-rich/necrotic core (LR/NC) (kappa=0.69; 0.31 to 0.86), almost perfect for hemorrhages (kappa=0.82; 0.68 to 0.96), and moderate (kappa=0.58; 0.27 to 0.88) and fair (kappa=0.33; 0.09 to 0.56) for fibrous cap identification on T2-weighted and TOF images, respectively. Interobserver agreement was substantial for the identification of calcifications (kappa=0.74; 0.59 to 0.89) and hemorrhages (kappa=0.62; 0.43 to 0.81), and moderate for LR/NC (kappa=0.58; 0.20 to 0.95). Agreement was fair for fibrous cap identification on both T2-weighted (kappa=0.28; -0.03 to 0.59) and on TOF images (kappa=0.26; 0.04 to 0.48). Agreement between T2 and TOF images for fibrous cap identification was slight (kappa=0.16; 0.01 to 0.31). Intra- and interobserver reproducibility for quantitative area measurements of vessel, lumen, plaque, LR/NC, and fibrous components was high with intraclass correlation coefficients ranging from 0.73 to 0.99. However, for the LR/NC, the interval delimited by the Bland-Altman graphs was wide in comparison to the mean. CONCLUSIONS: Vessel and plaque quantification is reproducible. Reproducibility of MRI for identifying and quantifying carotid plaque components is overall acceptable, but there is still significant variability that should be taken into account in the design of prognosis studies and clinical trials. Reproducibility for fibrous cap identification needs to be improved. PMID- 17463312 TI - Sex hormones and stroke rehabilitation in men and women. PMID- 17463313 TI - Infrared laser therapy for ischemic stroke: a new treatment strategy: results of the NeuroThera Effectiveness and Safety Trial-1 (NEST-1). AB - BACKGROUND AND PURPOSE: The NeuroThera Effectiveness and Safety Trial-1 (NEST-1) study evaluated the safety and preliminary effectiveness of the NeuroThera Laser System in the ability to improve 90-day outcomes in ischemic stroke patients treated within 24 hours from stroke onset. The NeuroThera Laser System therapeutic approach involves use of infrared laser technology and has shown significant and sustained beneficial effects in animal models of ischemic stroke. METHODS: This was a prospective, intention-to-treat, multicenter, international, double-blind, trial involving 120 ischemic stroke patients treated, randomized 2:1 ratio, with 79 patients in the active treatment group and 41 in the sham (placebo) control group. Only patients with baseline stroke severity measured by National Institutes of Health Stroke Scale (NIHSS) scores of 7 to 22 were included. Patients who received tissue plasminogen activator were excluded. Outcome measures were the patients' scores on the NIHSS, modified Rankin Scale (mRS), Barthel Index, and Glasgow Outcome Scale at 90 days after treatment. The primary outcome measure, prospectively identified, was successful treatment, documented by NIHSS. This was defined as a complete recovery at day 90 (NIHSS 0 to 1), or a decrease in NIHSS score of at least 9 points (day 90 versus baseline), and was tested as a binary measure (bNIH). Secondary outcome measures included mRS, Barthel Index, and Glasgow Outcome Scale. Primary statistical analyses were performed with the Cochran-Mantel-Haenszel rank test, stratified by baseline NIHSS score or by time to treatment for the bNIH and mRS. Logistic regression analyses were conducted to confirm the results. RESULTS: Mean time to treatment was >16 hours (median time to treatment 18 hours for active and 17 hours for control). Time to treatment ranged from 2 to 24 hours. More patients (70%) in the active treatment group had successful outcomes than did controls (51%), as measured prospectively on the bNIH (P=0.035 stratified by severity and time to treatment; P=0.048 stratified only by severity). Similarly, more patients (59%) had successful outcomes than did controls (44%) as measured at 90 days as a binary mRS score of 0 to 2 (P=0.034 stratified by severity and time to treatment; P=0.043 stratified only by severity). Also, more patients in the active treatment group had successful outcomes than controls as measured by the change in mean NIHSS score from baseline to 90 days (P=0.021 stratified by time to treatment) and the full mRS ("shift in Rankin") score (P=0.020 stratified by severity and time to treatment; P=0.026 stratified only by severity). The prevalence odds ratio for bNIH was 1.40 (95% CI, 1.01 to 1.93) and for binary mRS was 1.38 (95% CI, 1.03 to 1.83), controlling for baseline severity. Similar results held for the Barthel Index and Glasgow Outcome Scale. Mortality rates and serious adverse events (SAEs) did not differ significantly (8.9% and 25.3% for active 9.8% and 36.6% for control, respectively, for mortality and SAEs). CONCLUSIONS: The NEST-1 study indicates that infrared laser therapy has shown initial safety and effectiveness for the treatment of ischemic stroke in humans when initiated within 24 hours of stroke onset. A larger confirmatory trial to demonstrate safety and effectiveness is warranted. PMID- 17463314 TI - Has free radical release across the brain after carotid endarterectomy traditionally been underestimated? Significance of reperfusion hemodynamics. AB - BACKGROUND AND PURPOSE: Ischemia-reperfusion is an established paradigm for the induction of neuro-oxidative stress. The present report highlights the limitations associated with the measurement of free radical exchange across the human brain after carotid endarterectomy if reperfusion hemodynamics are not taken into account. Summary of Report- Only 2 human studies have reported local changes in the arterio-jugular bulb venous concentration difference (a-v(diff)) of free radicals during carotid endarterectomy. The authors reported either no change or only a very minor trans-cerebral release during the course of reperfusion, which was unexpected. However, consistent with other surgical models of ischemia-reperfusion, reperfusion would have been expected to increase plasma volume consistent with reflow-hemodilution. This would artifactually dilute the local concentration of free radicals, attenuate the a-v(diff) and thus underestimate the "true" magnitude of cerebral free radical release. CONCLUSIONS: After correction for reflow-hemodilution, the cerebral generation of free radicals after carotid endarterectomy is likely to be significantly more pronounced than previously documented in humans. PMID- 17463315 TI - New pediatric model of ischemic stroke in infant piglets by photothrombosis: acute changes in cerebral blood flow, microvasculature, and early histopathology. AB - BACKGROUND AND PURPOSE: The etiology and pathophysiology of acute ischemic stroke in children differ greatly from those in adults. The purpose of this study was to establish a new pediatric model of ischemic stroke in infant piglets for use in future studies of the response of the developing brain to focal ischemic injury. METHODS: Ischemic stroke was produced in male infant piglets (2 to 4 weeks old) by photothrombotic occlusion of the middle cerebral artery. Regional cerebral blood flow was measured with radiolabeled microspheres up to 4 hours after occlusion. Early histopathology, including caspase-3 immunohistochemistry for apoptosis, was examined 4 hours after ischemia. The nature of the thrombus and its interaction with vascular endothelium were assessed by electron microscopy. RESULTS: Severe ischemia (0 to 15 mL/100 g per min) occurred rapidly in 1.4+/-0.2 g of tissue at 15 minutes and increased to 2.4+/-0.7 g at 4 hours. Similarly, moderate ischemia (16 to 30 mL/100 g per min) was measured in 1.2+/-0.3 g of tissue at 15 minutes and increased to 2.0+/-0.6 g at 4 hours. These regional cerebral blood flow values represent ischemic levels of blood flow in 20% to 25% of the volume of the ischemic hemisphere at 4 hours after ischemia. Ischemic infarction occurred in both gray and white matter, and cerebral microvessels in the ischemic hemisphere contained large numbers of inflammatory leukocytes. Caspase-3-positive cells were few in number and were found in the periphery of the infarct; cell death appeared to occur primarily by necrosis rather than apoptosis at 4 hours. Electron microscopy revealed a pure platelet thrombus firmly attached to the vascular endothelium, which in some areas appeared to be detached from the basement membrane. CONCLUSIONS: Ischemic stroke can be produced in infant piglets by middle cerebral artery photothrombosis. The stroke involved both gray and white matter and exhibited a robust inflammatory component. The mean infarct volume determined histopathologically amounted to 9.6+/-2.4% of the affected (ipsilateral) hemisphere, which was correlated well with the mass equivalent of tissue (12.0+/-3.5%), in which severe declines in regional cerebral blood flow were observed at 4 hours. PMID- 17463316 TI - Can we improve the statistical analysis of stroke trials? Statistical reanalysis of functional outcomes in stroke trials. AB - BACKGROUND AND PURPOSE: Most large acute stroke trials have been neutral. Functional outcome is usually analyzed using a yes or no answer, eg, death or dependency versus independence. We assessed which statistical approaches are most efficient in analyzing outcomes from stroke trials. METHODS: Individual patient data from acute, rehabilitation and stroke unit trials studying the effects of interventions which alter functional outcome were assessed. Outcomes included modified Rankin Scale, Barthel Index, and "3 questions". Data were analyzed using a variety of approaches which compare 2 treatment groups. The results for each statistical test for each trial were then compared. RESULTS: Data from 55 datasets were obtained (47 trials, 54,173 patients). The test results differed substantially so that approaches which use the ordered nature of functional outcome data (ordinal logistic regression, t test, robust ranks test, bootstrapping the difference in mean rank) were more efficient statistically than those which collapse the data into 2 groups (chi(2); ANOVA, P<0.001). The findings were consistent across different types and sizes of trial and for the different measures of functional outcome. CONCLUSIONS: When analyzing functional outcome from stroke trials, statistical tests which use the original ordered data are more efficient and more likely to yield reliable results. Suitable approaches included ordinal logistic regression, t test, and robust ranks test. PMID- 17463317 TI - Meta-analysis of traditional Chinese patent medicine for ischemic stroke. AB - BACKGROUND AND PURPOSE: A large number of traditional Chinese patent medicine (TCPM) are widely used for ischemic stroke in China. The aim of this study was to systematically review the existing clinical evidence on TCPM for ischemic stroke. METHODS: We identified all TCPM that were listed in the Chinese National Essential Drug list of 2004 and those commonly used TCPM in current clinical practice for ischemic stroke. Fifty-nine TCPM were identified for further evaluation. We applied Cochrane systematic review methods. We searched for reports of randomized controlled trials and controlled clinical trials on any of the 59 TCPM for ischemic stroke comparing one TCPM with control. PRIMARY OUTCOMES included death or dependency at the end of follow-up (at least 3 months) and adverse events. Effects on neurological impairments were a secondary outcome. RESULTS: One-hundred ninety-one trials (19,338 patients) on 22 TCPM were available and included, of which 120 were definite or possible randomized controlled trials and 71 were controlled clinical trials. The methodological quality of included trials was generally "poor." Few trials reported methods of randomization. Three trials were randomized, double blind, and placebo controlled. PRIMARY OUTCOMES: one trial on Puerarin and one trial on Shenmai injection assessed death or dependency at the end of long-term follow-up (at least 3 months) and found no statistically significant difference between 2 groups. The reported adverse events including allergic reaction, headache, nausea, diarrhea, bellyache, blood pressure change, and subcutaneous ecchymosis. Most of the adverse events were not severe. SECONDARY OUTCOMES: analysis of the secondary outcome, "marked improvement in neurological deficit," showed apparent benefits of about the same magnitude for all the TCPM studied. Of the 22 TCPM, 8 drugs (Milk vetch, Mailuoning, Ginkgo biloba, Ligustrazine, Danshen agents, Xuesetong, Puerarin, and Acanthopanax) had relatively more studies and patient numbers. CONCLUSIONS: There was insufficient good quality evidence on the effects of TCPM in ischemic stroke on the primary outcome (death or dependency). We considered the apparent benefit on neurological impairment was as likely to be attributable to bias from poor methodology as to a real treatment effect. However, because the agents assessed appeared potentially beneficial and nontoxic, further randomized controlled trials are justified. Eight drugs could be further research priorities. PMID- 17463318 TI - Size matters: hemorrhage volume as an objective measure to define significant intracranial hemorrhage associated with thrombolysis. AB - BACKGROUND AND PURPOSE: This study defines significant thrombolysis associated intracranial hemorrhage (ICH) by identifying an objective threshold volume that predicts clinical deterioration attributable to ICH. METHODS: Prospectively collected clinical and radiographic information, from 103 consecutive patients who underwent intraarterial thrombolysis for acute ischemic stroke, was reviewed. Multiple paired comparisons between stratified hematoma volume and change in National Institutes of Health Stroke Scale (NIHSS) score by 24 to 36 hours and by time of hospital discharge was used to identify significant differences. Associations between hemorrhage volume and infarct volume in relation to clinical outcomes were examined. Rates of hemorrhagic transformation (HT), symptomatic hemorrhage, and parenchymal hematoma involving over 30% of the infarct were compared with hemorrhage volume. Multivariate regression analysis was used to determine the relationship between change in discharge NIHSS score and hemorrhage volume adjusting for known predictors of clinical outcomes. RESULTS: Multiple paired comparisons indicate that hemorrhage greater than 25 mL (HV25) had a more distinct impact on NIHSS score by time of hospital discharge than at 24 to 36 hours. Twenty-seven (26.2%) patients had HT and 12 (11.7%) had HV25. Among symptomatic hemorrhage, parenchymal hematoma involving over 30% of the infarct, and HV25, HV25 appeared more reflective of clinical deterioration from ICH. Hemorrhage volume increased with infarct volume but they were independently associated with change in NIHSS score on regression analysis. CONCLUSIONS: Clinical deterioration from ICH and ischemic injury are more effectively distinguished at time of hospital discharge. The authors propose to define significant hemorrhage associated with thrombolysis as hemorrhage volume greater than 25 mL. PMID- 17463319 TI - Angiotensin II increases expression of alpha1C subunit of L-type calcium channel through a reactive oxygen species and cAMP response element-binding protein dependent pathway in HL-1 myocytes. AB - Angiotensin II (Ang II) is involved in the pathogenesis of atrial fibrillation (AF). L-type calcium channel (LCC) expression is altered in AF remodeling. We investigated whether Ang II modulates LCC current through transcriptional regulation, by using murine atrial HL-1 cells, which have a spontaneous calcium transient, and an in vivo rat model. Ang II increased LCC alpha1C subunit mRNA and protein levels and LCC current density, which resulted in an augmented calcium transient in atrial myocytes. An approximately 2-kb promoter region of LCC alpha1C subunit gene was cloned to the pGL3 luciferase vector. Ang II significantly increased promoter activity in a concentration- and time-dependent manner. Truncation and mutational analysis of the LCC alpha1C subunit gene promoter showed that cAMP response element (CRE) (-1853 to -1845) was an important cis element in Ang II-induced LCC alpha1C subunit gene expression. Transfection of dominant-negative CRE binding protein (CREB) (pCMV-CREBS133A) abolished the Ang II effect. Ang II (1 micromol/L, 2 hours) induced serine 133 phosphorylation of CREB and binding of CREB to CRE and increased LCC alpha1C subunit gene promoter activity through a protein kinase C/NADPH oxidase/reactive oxygen species pathway, which was blocked by the Ang II type 1 receptor blocker losartan and the antioxidant simvastatin. In the rat model, Ang II infusion increased LCC alpha1C subunit expression and serine 133 phosphorylation of CREB, which were attenuated by oral losartan and simvastatin. In summary, Ang II induced LCC alpha1C subunit expression via a protein kinase C-, reactive oxygen species-, and CREB-dependent pathway and was blocked by losartan and simvastatin. PMID- 17463320 TI - Thin filament disinhibition by restrictive cardiomyopathy mutant R193H troponin I induces Ca2+-independent mechanical tone and acute myocyte remodeling. AB - Inherited restrictive cardiomyopathy (RCM) is a debilitating disease characterized by a stiff heart with impaired ventricular relaxation. Mutations in cardiac troponin I (cTnI) were identified as causal for RCM. Acute genetic engineering of adult cardiac myocytes was used to identify primary structure/function effects of mutant cTnI. Studies focused on R193H cTnI owing to the poor prognosis of this allele. Compared with wild-type cTnI, R193H mutant cTnI more effectively incorporated into the sarcomere, where it exerted dose dependent effects on basal and dynamic contractile function. Under loaded conditions, permeabilized myocyte Ca(2+) sensitivity of tension was increased, whereas the passive tension-extension relationship was not altered by R193H cTnI. Normal rod-shaped myocyte morphology acutely transitioned to a "short-squat" phenotype in concert with progressive stoichiometric incorporation of R193H in the absence of altered diastolic Ca(2+). The specific myosin inhibitor blebbistatin fully blocked this transition. Heightened Ca(2+) buffering by the R193H myofilaments, and not alterations in Ca(2+) handling by the sarcoplasmic reticulum, slowed the decay rate of the Ca(2+) transient. Incomplete mechanical relaxation conferred by R193H was exacerbated at increasing pacing frequencies independent of elevated diastolic Ca(2+). R193H cTnI-dependent mechanical tone caused acute remodeling to a quasicontracted state not elicited by other Ca(2+) sensitizing proteins and is a direct correlate of the stiff heart characteristic of RCM in vivo. These results point toward targets downstream of Ca(2+) handling, notably thin filament regulation and actin-myosin interaction, in designing therapeutic strategies to redress the primary cell morphological and mechanical underpinnings of RCM. PMID- 17463321 TI - Statins activate peroxisome proliferator-activated receptor gamma through extracellular signal-regulated kinase 1/2 and p38 mitogen-activated protein kinase-dependent cyclooxygenase-2 expression in macrophages. AB - Both statins and peroxisome proliferator-activated receptor (PPAR)gamma ligands have been reported to protect against the progression of atherosclerosis. In the present study, we investigated the effects of statins on PPARgamma activation in macrophages. Statins increased PPARgamma activity, which was inhibited by mevalonate, farnesylpyrophosphate, or geranylgeranylpyrophosphate. Furthermore, a farnesyl transferase inhibitor and a geranylgeranyl transferase inhibitor mimicked the effects of statins. Statins inhibited the membrane translocations of Ras, RhoA, Rac, and Cdc42, and overexpression of dominant-negative mutants of RhoA (DN-RhoA) and Cdc42 (DN-Cdc42), but not of Ras or Rac, increased PPARgamma activity. Statins induced extracellular signal-regulated kinase (ERK)1/2 and p38 mitogen-activated protein kinase (MAPK) activation. However, DN-RhoA and DN-Cdc42 activated p38 MAPK, but not ERK1/2. ERK1/2- or p38 MAPK-specific inhibitors abrogated statin-induced PPARgamma activation. Statins induced cyclooxygenase (COX)-2 expression and increased intracellular 15-deoxy-Delta(12,14) prostaglandin J(2) (15d-PGJ(2)) levels through ERK1/2- and p38 MAPK-dependent pathways, and inhibitors or small interfering RNA of COX-2 inhibited statin induced PPARgamma activation. Statins also activate PPARalpha via COX-2-dependent increases in 15d-PGJ(2) levels. We further demonstrated that statins inhibited lipopolysaccharide-induced tumor necrosis factor alpha or monocyte chemoattractant protein-1 mRNA expression, and these effects by statins were abrogated by the PPARgamma antagonist T0070907 or by small interfering RNA of PPARgamma or PPARalpha. Statins also induced ATP-binding cassette protein A1 or CD36 mRNA expression, and these effects were suppressed by small interfering RNAs of PPARgamma or PPARalpha. In conclusion, statins induce COX-2-dependent increase in 15d-PGJ(2) level through a RhoA- and Cdc42-dependent p38 MAPK pathway and a RhoA- and Cdc42-independent ERK1/2 pathway, thereby activating PPARgamma. Statins also activate PPARalpha via COX-2-dependent pathway. These effects of statins may explain their antiatherogenic actions. PMID- 17463322 TI - Exact assessment of perfusion and collateral vessel proliferation in small animal models. PMID- 17463323 TI - Patient and cellular characteristics determine efficacy of cell therapy. PMID- 17463324 TI - Human PON3, effects beyond the HDL: clues from human PON3 transgenic mice. PMID- 17463325 TI - Therapeutic challenge to adiposity of the heart. PMID- 17463326 TI - G proteins: more than transducers of receptor-generated signals? PMID- 17463327 TI - Stem cells and the regeneration of the aging cardiovascular system. AB - It is well established that cardiovascular repair mechanisms become progressively impaired with age and that advanced age is itself a significant risk factor for cardiovascular disease. Although therapeutic developments have improved the prognosis for those with cardiovascular disease, mortality rates have nevertheless remained virtually unchanged in the last twenty years. Clearly, there is a need for alternative strategies for the treatment of cardiovascular disease. In recent years, the idea that the heart is capable of regeneration has raised the possibility that cell-based therapies may provide such an alternative to conventional treatments. Cells that have the potential to generate cardiomyocytes and vascular cells have been identified in both the adult heart and peripheral tissues, and in vivo experiments suggest that these cardiovascular stem cells and cardiovascular progenitor cells, including endothelial progenitor cells, are capable of replacing damaged myocardium and vascular tissues. Despite these findings, the endogenous actions of cardiovascular stem cells and cardiovascular progenitor cells appear to be insufficient to protect against cardiovascular disease in older individuals. Because recent evidence suggests that cardiovascular stem cells and cardiovascular progenitor cells are subject to age-associated changes that impair their function, these changes may contribute to the dysregulation of endogenous cardiovascular repair mechanisms in the aging heart and vasculature. Here we present the evidence for the impact of aging on cardiovascular stem cell/cardiovascular progenitor cell function and its potential importance in the increased severity of cardiovascular pathophysiology observed in the geriatric population. PMID- 17463328 TI - Endothelial mitochondria: contributing to vascular function and disease. AB - Disturbances in vascular function contribute to the development of several diseases of increasing prevalence and thereby contribute significantly to human mortality and morbidity. Atherosclerosis, diabetes, heart failure, and ischemia with attendant reperfusion injury share many of the same risk factors, among the most important being oxidative stress and alterations in the blood concentrations of compounds that influence oxidative stress, such as oxidized low-density lipoprotein. In this review, we focus on endothelial cells: cells in the frontline against these disturbances. Because ATP supplies in endothelial cells are relatively independent of mitochondrial oxidative pathways, the mitochondria of endothelial cells have been somewhat neglected. However, they are emerging as agents with diverse roles in modulating the dynamics of intracellular calcium and the generation of reactive oxygen species and nitric oxide. The mitochondria may also constitute critical "targets" of oxidative stress, because survival of endothelial cells can be compromised by opening of the mitochondrial permeability transition pore or by mitochondrial pathways of apoptosis. In addition, evidence suggests that endothelial mitochondria may play a "reconnaissance" role. For example, although the exact mechanism remains obscure, endothelial mitochondria may sense levels of oxygen in the blood and relay this information to cardiac myocytes as well as modulating the vasodilatory response mediated by endothelial nitric oxide. PMID- 17463329 TI - Seven-transmembrane receptors and ubiquitination. AB - Regulation of protein function by posttranslational modification plays an important role in many biological pathways. The most well known among such modifications is protein phosphorylation performed by highly specific protein kinases. In the past decade, however, covalent linkage of the low-molecular weight protein ubiquitin to substrate proteins (protein ubiquitination) has proven to be yet another widely used mechanism of protein regulation playing a crucial role in virtually all aspects of cellular functions. This review highlights some of the recently discovered and provocative roles for ubiquitination in the regulation of the life cycle and signal transduction properties of 7-transmembrane receptors that serve to integrate many biological functions and play fundamental roles in cardiovascular homeostasis. PMID- 17463330 TI - Inducible cAMP early repressor inhibits growth of vascular smooth muscle cell. AB - OBJECTIVE: The role of inducible cAMP early repressor (ICER), a transcriptional repressor, in the vascular remodeling process has not been determined. We examined whether ICER affects growth of vascular smooth muscle cells (VSMCs). METHODS AND RESULTS: Semi-quantitative RT-PCR and Western blot analysis showed that expression of ICER was increased in beraprost (a prostaglandin I2 analogue) stimulated VSMCs in a time- and dose-dependent manner. The induction of ICER was inhibited by pretreatment with H89, a protein kinase A (PKA) inhibitor, suggesting that PKA mediates the induction of ICER expression. Beraprost suppressed platelet-derived growth factor-induced thymidine incorporation in VSMCs, which was reversed by transfection of short interfering RNA for ICER, not by scramble RNA. Overexpression of ICER by an adenovirus vector attenuated neointimal formation (intima/media ratio) by 50% compared with overexpression of LacZ. The number of terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling-positive cells was increased and the number of Ki-67-positive cells was decreased in ICER-transduced artery. CONCLUSION: These results suggest that ICER induces apoptosis and inhibits proliferation of VSMCs, and plays a critical role in beraprost-mediated suppression of VSMC proliferation. ICER may be an important endogenous inhibitor of vascular proliferation. PMID- 17463331 TI - Endothelial fibrinolytic capacity predicts future adverse cardiovascular events in patients with coronary heart disease. AB - OBJECTIVE: The endothelium-derived fibrinolytic factor tissue plasminogen activator (t-PA) is a major determinant of vessel patency after coronary plaque rupture and thrombosis. We assessed whether endothelial fibrinolytic capacity predicts atherothrombotic events in patients with coronary heart disease. METHODS AND RESULTS: Plasma t-PA and plasminogen activator inhibitor (PAI)-1 concentrations were measured during intrabrachial substance P infusion in 98 patients with angiographically proven stable coronary heart disease. Forearm blood flow was measured during infusion of substance P and sodium nitroprusside. Cardiovascular events (cardiovascular death, myocardial infarction [MI], ischemic stroke [CVA], and emergency hospitalization for unstable angina) were determined during 42 months of follow-up. Patients experiencing a cardiovascular event (n=19) had similar baseline characteristics to those free of events. Substance P caused a dose-dependent increase in plasma t-PA concentrations (P<0.001). However, net t-PA release was 72% lower in the patients who experienced death, MI, or CVA, and 48% lower in those who suffered death, MI, CVA or hospitalization for unstable angina (P<0.05). Major adverse cardiovascular events were most frequent in those with the lowest fibrinolytic capacity (P=0.03 for trend); patients with the lowest quartile of t-PA release had the highest rate of adverse events (P=0.01). CONCLUSION: Endothelial fibrinolytic capacity, as measured by stimulated t-PA release, predicts the future risk of adverse cardiovascular events in patients with coronary heart disease. We suggest that endothelial fibrinolytic capacity is a powerful novel determinant of cardiovascular risk. PMID- 17463332 TI - Venous identity is lost but arterial identity is not gained during vein graft adaptation. AB - OBJECTIVES: Ephrin ligands and Eph receptors are signaling molecules that are differentially expressed on arteries and veins during development. We examined whether Eph-B4, a venous marker, and Ephrin-B2, an arterial marker, are regulated during vein graft adaptation in humans and aged rats. METHODS AND RESULTS: Eph-B4 transcripts and immunodetectable protein are downregulated in endothelial and smooth muscle cells of patent vein grafts in both humans and in aged rats, whereas Ephrin-B2 transcripts and protein are not strongly induced. Other markers of arterial identity, including dll4 and notch-4, are also not induced during vein graft adaptation in aged rats. Because VEGF-A is upstream of the Ephrin-Eph pathway, and expression of VEGF-A is induced only at early time points after exposure of the vein to the arterial environment, we inhibited VEGF-A in vein grafts using an siRNA-based approach. Vein grafts treated with siRNA directed against VEGF-A demonstrated a thicker intima-media containing alpha-actin, consistent with arterialization, but did not contain Eph-B4 or Ephrin-B2. CONCLUSIONS: Venous identity is preserved in the veins of aged animals, but is lost during adaptation to the arterial circulation; arterial markers are not induced. Markers of vessel identity are plastic in adults and their selective regulation may mediate vein graft adaptation to the arterial environment in aged animals and humans. PMID- 17463333 TI - A specific role for eNOS-derived reactive oxygen species in atherosclerosis progression. AB - OBJECTIVE: When the availability of tetrahydrobiopterin (BH4) is deficient, endothelial nitric oxide synthase (eNOS) produces superoxide rather than NO (uncoupled eNOS). We have shown that the atherosclerotic lesion size was augmented in apolipoprotein E-deficient (ApoE-KO) mice overexpressing eNOS because of the enhanced superoxide production. In this study, we addressed the specific importance of uncoupled eNOS in atherosclerosis, and the potential mechanistic role for specific versus nonspecific antioxidant strategies in restoring eNOS coupling. METHODS AND RESULTS: We crossed mice overexpressing eNOS in the endothelium (eNOS-Tg) with mice overexpressing GTP-cyclohydrolase I (GCH), the rate-limiting enzyme in BH4 synthesis, to generate ApoE-KO/eNOS-Tg/GCH-Tg mice. As a comparison, ApoE-KO/eNOS-Tg mice were treated with vitamin C. Atherosclerotic lesion formation was increased in ApoE-KO/eNOS-Tg mice compared with ApoE-KO mice. GCH overexpression in ApoE-KO/eNOS-Tg/GCH-Tg mice increased vascular BH4 levels and reduced plaque area. This reduction was associated with decreased superoxide production from uncoupled eNOS. Vitamin C treatment failed to reduce atherosclerotic lesion size in ApoE-KO/eNOS-Tg mice, despite reducing overall vascular superoxide production. CONCLUSION: In contrast to vitamin C treatment, augmenting BH4 levels in the endothelium by GCH overexpression reduced the accelerated atherosclerotic lesion formation in ApoE-KO/eNOS-Tg mice, associated with a reduction of superoxide production from uncoupled eNOS. PMID- 17463334 TI - Platelet receptor proteolysis: a mechanism for downregulating platelet reactivity. AB - The platelet plasma membrane is literally at the cutting-edge of recent research into proteolytic regulation of the function and surface expression of platelet receptors, revealing new mechanisms for how the thrombotic propensity of platelets is controlled in health and disease. Extracellular proteolysis of receptors irreversibly inactivates receptor-mediated adhesion and signaling, as well as releasing soluble fragments into the plasma where they act as potential markers or modulators. Platelet-surface sheddases, particularly of the metalloproteinase-disintegrin (ADAM) family, can be regulated by many of the same mechanisms that control receptor function, such as calmodulin association or activation of signaling pathways. This provides layers of regulation (proteinase and receptor), and a higher order of control of cellular function. Activation of pathways leading to extracellular shedding is concomitant with activation of intracellular proteinases such as calpain, which may also irreversibly deactivate receptors. In this review, platelet receptor shedding will be discussed in terms of (1) the identity of proteinases involved in receptor proteolysis, (2) key platelet receptors regulated by proteolytic pathways, and (3) how shedding might be regulated in normal physiology or future therapeutics. In particular, a focus on proteolytic regulation of the platelet collagen receptor, glycoprotein (GP)VI, illustrates many of the key biochemical, cellular, and clinical implications of current research in this area. PMID- 17463335 TI - Impact of serum amyloid A on tissue factor and tissue factor pathway inhibitor expression and activity in endothelial cells. AB - OBJECTIVE: Although serum amyloid A (SAA) is a useful biomarker of coronary artery disease (CAD), its direct role in procoagulation is obscure. This study investigates the impact of SAA on the expression and activity of tissue factor (TF) and tissue factor pathway inhibitor (TFPI) in endothelial cells. METHODS AND RESULTS: SAA was found to disturb the balance of TF and TFPI expression and activity in human endothelial cells. SAA (20 microg/mL) markedly induced TF expression between 4 to 8 hours in both protein and mRNA levels, as well as TF activity. Conversely, incubation of SAA (20 microg/mL) for 24 and 48 hours was found to significantly inhibit TFPI secretion, transcription, and activity. Pretreatment with formyl peptide receptor-like 1 (FPRL1) inhibitors (Pertussis toxin and WRWWWW) could block the SAA effects on TF and TFPI. Furthermore, pretreatment with the respective specific mitogen-activated protein kinase (MAPK) inhibitors (SB203580, PD98059, and SP600125) and NFkappaB inhibitor (Bay-11 to 7082) could block SAA-dependent TF induction. SAA also directly induced activation of MAP kinases and NFkappaB. CONCLUSIONS: The stimulating effect of SAA was faster-acting on the expression and activity of TF and the inhibitory effect was slower-acting on TFPI. The effects are mediated through FPRL1, MAP kinases and NFkappaB. PMID- 17463336 TI - Improving the quality of care for children: let's dance. PMID- 17463337 TI - Psychiatrists and primary caring: what are our boundaries of responsibility? AB - This column provides a framework for considering the extent of psychiatrists' responsibility for patients' medical conditions. Psychiatrists have the greatest responsibility for medical conditions that occur as a result of their own actions. Next on the continuum of responsibility is psychiatrists' obligation to remain alert for medical conditions that can cause, trigger, or exacerbate psychiatric conditions or interfere with treatment. Another potential responsibility is for preventive monitoring, screening, and education for medical conditions that disproportionately affect psychiatric patients. Characteristics of the setting, practitioner, and patient that affect how such responsibilities are fulfilled are also discussed. PMID- 17463338 TI - School characteristics and behavior problems of U.S. fifth-graders. PMID- 17463339 TI - Culture, children, and mental health treatment: special section on the national stigma study-children. PMID- 17463340 TI - Stigmatizing attitudes and beliefs about treatment and psychiatric medications for children with mental illness. AB - OBJECTIVES: Data on community responses to two treatment issues critical for children and adolescents with mental health problems are addressed: stigma associated with receiving mental health care and the willingness to use psychiatric medication. METHODS: Using a representative sample of the U.S. population, the investigators interviewed 1,393 noninstitutionalized adult participants in the National Stigma Study-Children (NSS-C) (response rate 70%; margin of error +/- 4%). RESULTS: Many respondents believed that stigma results from mental health treatment during childhood (45% reported likely rejection at school) and that stigma continues to have negative ramifications into adulthood (43%). More than half (57%) were skeptical about confidentiality, and more than one-third (35%) expected parents of children with mental illness to experience self-stigma. Most respondents believed that psychiatric medications affect development (68%), give children a flat, "zombie"-like affect (53%), and delay solving "real" behavior-related problems (66%). Most (86%) believed that physicians overmedicate children for common behavioral problems. Women and persons with more education rather than less perceived less stigma resulting from treatment but reported more negative views on medication. Beliefs in medication efficacy and trust in physicians were associated with more positive cultural beliefs. CONCLUSIONS: Data on the contemporary cultural context surrounding treatment for children's mental health issues revealed substantial stigma concerns, particularly surrounding medication options. These beliefs and attitudes cannot be easily inferred from adults' sociodemographic characteristics. PMID- 17463341 TI - Perceived dangerousness of children with mental health problems and support for coerced treatment. AB - OBJECTIVE: This study examined the public's beliefs regarding the potential for harm to self and others and the public's willingness to invoke coercive or legal means to ensure treatment of children. METHODS: Using data from the National Stigma Study-Children (NSS-C), which presented vignettes to 1,152 individuals, the investigators compared public perceptions of the dangerousness of children with attention-deficit hyperactivity disorder (ADHD), major depression, asthma, and "daily troubles." Multivariate analyses were used to examine the predictors of perceptions of dangerousness and the willingness to support legally enforced treatment of these conditions. RESULTS: Children with ADHD and children with major depression were perceived (by 33% and 81% of the sample, respectively) as somewhat likely or very likely to be dangerous to themselves or others, compared with children with asthma (15%) or those with "daily troubles" (13%). Over one third of the sample (35%) were willing to use legal means to force children with depression to see a clinician. However, even more (42%) endorsed forced treatment for a child with asthma. Furthermore, individuals who labeled the child as "mentally ill" were approximately twice as likely to report a potential for violence and five times as likely to support forced treatment. CONCLUSIONS: Large numbers of people in the United States link children's mental health problems, particularly depression, to a potential for violence and support legally mandated treatment. These evaluations appear to reflect the stigma associated with mental illness and the public's concern for parental responsibility. PMID- 17463342 TI - Public knowledge, beliefs, and treatment preferences concerning attention-deficit hyperactivity disorder. AB - OBJECTIVE: This study aimed to understand the level of public knowledge about attention-deficit hyperactivity disorder (ADHD), treatment preferences for the disorder, and their sociodemographic correlates. METHODS: A short battery of questions about ADHD was included in the 2002 General Social Survey (N=1,139). In face-to-face interviews, respondents answered questions about whether they had heard of ADHD, what they knew about ADHD, their beliefs about whether ADHD is a "real" disease, and opinions about whether children with ADHD should be offered counseling or medication. RESULTS: Just under two-thirds of respondents (64%) had heard of ADHD; most could not provide detailed information about the disorder. Women and those with higher levels of education were more likely to have heard of ADHD; African Americans, members of other nonwhite racial and ethnic groups, and older respondents were less likely to have heard of ADHD. Among respondents who had heard of ADHD, 78% said they believed ADHD to be a real disease; women, white respondents, and persons with higher income most often endorsed that belief. Most respondents (65%) endorsed the use of both counseling and medication, although counseling was endorsed as a sole treatment more often than medication. There were few sociodemographic differences in treatment preferences. CONCLUSIONS: The public is not well informed about ADHD. Future media and educational efforts should seek to provide accurate information about ADHD, with a special effort to reach specific populations such as men, nonwhite minority groups, and older Americans. PMID- 17463343 TI - Comparison of public attributions, attitudes, and stigma in regard to depression among children and adults. AB - OBJECTIVE: This study compared public attributions and attitudes toward adult and child depression, with a focus on problem recognition, medical and social causes, help-seeking recommendations, perceptions of violence, and the use of coercion. METHODS: The investigators compared data from two special modules of the 1996 and 2002 nationally representative General Social Survey on public response to mental illness. Respondents answered questions regarding a vignette in which an adult had depression (N=193) or one in which a child had depression (N=312). RESULTS: Respondents evaluated childhood depression as more serious than adult depression (83% versus 51%, respectively) and saw a greater potential for violence toward others among children with depression (40% for children versus 30% for adults). More respondents endorsed treatment of all types, including coerced care, for children with depression. However, significantly fewer recommended talking to family and friends about a child's mental health problem. CONCLUSIONS: Americans are more concerned about children's depression than adults' depression and reveal more prejudice regarding perceptions of dangerousness. More respondents endorsed formal care than informal care and advice. However, the heightened stigma surrounding childhood depression poses unique challenges for youths with depression and their families. PMID- 17463344 TI - Coming to terms with ADHD: how urban African-American families come to seek care for their children. AB - OBJECTIVE: This study investigated how parents' interpretations of their child's disruptive or inattentive behaviors led them to seek medical care that resulted in a diagnosis of attention-deficit hyperactivity disorder (ADHD). METHODS: Qualitative, semistructured telephone interviews were conducted with parents of children who had been newly diagnosed as having ADHD (96% of the children were African American). The 26 participants were recruited from primary care, developmental and behavioral, and specialty mental health pediatric clinics affiliated with a large, urban teaching hospital. The analysis followed a grounded theory approach. RESULTS: By the time that parents sought treatment for their child's ADHD, they had been through an extensive process to pinpoint their child's problems. Parents' conceptualizations emerged as they described their child's behavior, explained the situation, described how ADHD affected their children, and explained how they sought answers. Their reactions to the behavior and visions they had for their child's future reflected their motivation to manage the situation. Parents' conceptualization and management of the behaviors described the process of coming to terms with their child's ADHD and the need for care. Four distinct patterns describing this process emerged from the analysis: immediate resolution, pragmatic management, attributional ambivalence, and coerced conformance. CONCLUSIONS: It is likely that clinicians' awareness of the different approaches by which families come to the decision to seek care for their child's ADHD will allow clinicians to provide more responsive care and better tailor interventions to improve therapeutic outcomes for children receiving mental health treatments. PMID- 17463345 TI - Americans' attitudes toward mental health treatment seeking: 1990-2003. AB - OBJECTIVE: This study examined recent trends in Americans' attitudes toward mental health treatment seeking and beliefs about the effectiveness of such treatment. METHODS: Data on attitudes and beliefs from two representative surveys of the U.S. general population were compared. Samples included 5,388 participants from the National Comorbidity Survey (NCS) in 1990-1992 and 4,319 from the National Comorbidity Survey-Replication (NCS-R) in 2001-2003. RESULTS: Participants in the 2001-2003 survey were more willing than those in the 1990 1992 survey to seek professional help for mental health problems (41.4% reported that they would "definitely go" for professional help in 2001-2003, compared with 35.6% in 1990-1992). Participants in the more recent survey were also more comfortable talking with a professional about personal problems (32.4% in 2001 2003 reported feeling "very comfortable," compared with 27.1% in 1990-1992) and were less likely to say that they would be embarrassed if others found out about it (40.3% reported being "not at all embarrassed" in 2001-2003, compared with 33.7% in 1990-1992). Attitudes of younger participants improved more than attitudes of middle-aged participants. Public beliefs about the effectiveness of mental health treatment and the likelihood of recovery without treatment changed little across surveys. CONCLUSIONS: Mental health treatment seeking has become more acceptable over the past decade, and perceived stigma associated with it has declined. These changes in public attitudes have likely contributed to the growing demand for mental health services in the United States and will continue to do so in the coming years. PMID- 17463346 TI - Association of chronic work stress, psychiatric disorders, and chronic physical conditions with disability among workers. AB - OBJECTIVE: There appear to be links between psychiatric disorders and work related stress as well as between psychiatric disorders and physical conditions. This study explores the relationships between chronic work stress, psychiatric disorders, and chronic physical conditions and disability among workers. By doing so, this study sought to understand how these factors are associated with worker disability when they are experienced alone versus in combination with one another. METHODS: The study population was drawn from the Canadian Community Health Survey 1.2, a national population-based survey that gathered cross sectional data on health status from 22,118 working respondents. The relationship between chronic work stress, chronic physical conditions, and psychiatric disorders and disability in the past 14 days was examined for working respondents by using logistic regressions controlling for sociodemographic characteristics, region, and occupation. RESULTS: Thirty-one percent of respondents experienced chronic work stress either alone or in combination with a chronic physical condition, a psychiatric disorder, or both. Forty-six percent reported at least one chronic physical condition either alone or in combination. Finally, 11% had a psychiatric disorder. Compared with the group with none of the factors, those with an increasing number of combined conditions had increasing odds of disability after the analysis controlled for sociodemographic characteristics, occupation, and region. CONCLUSIONS: The presence of chronic work stress seems to amplify effects of psychiatric disorders and chronic physical conditions on disability. In addition, psychiatric disorders co-occurring with physical illness seem to be associated with significantly higher odds of disability. PMID- 17463347 TI - Association of mood, anxiety, and substance use disorders with occupational status and disability in a community sample. AB - OBJECTIVE: This study examined associations between mood disorders, anxiety disorders, substance dependence or harmful alcohol use, and occupational status and disability in a general population sample. METHODS: Data from the Canadian Community Health Survey Cycle 1.2-Mental Health and Well-Being (CCHS-1.2), a representative cross-sectional survey, were analyzed. The total sample was narrowed to individuals between the ages of 18 and 64 years, the age range most likely to be working. RESULTS: Of the 27,332 persons surveyed, 946 had a mood disorder only, 831 had an anxiety disorder only, 730 had substance dependence only, and 966 had more than one disorder. Twenty-three percent reported that during the previous week they were not at a job or were permanently unable to work (27% with mood disorder only, 30% with anxiety disorder only, and 20% with substance dependence only, and 34% with more than one disorder). In unadjusted analyses, mood and anxiety disorders were associated with absence from work during the week preceding the interview, whereas substance dependence was not. After adjustment for other variables using logistic regression, an association of substance dependence and work absence emerged. Each category of disorder was strongly associated with a greater likelihood of disability days or days spent in bed for mental health reasons. CONCLUSIONS: On a population level, mood and anxiety disorders and substance dependence were associated both with not working during the week preceding the interview as well as an increase in reported disability or bed days. The strength of association appears to be stronger for mood and anxiety disorders. PMID- 17463348 TI - Unintended consequences of regionalizing specialized VA addiction services. AB - OBJECTIVE: From 1995 to 2000 the Department of Veterans Affairs (VA) dramatically reduced addiction treatment funding and regionalized specialized services to urban centers. By using New York State as an example, this study examined whether regionalization disproportionately affected rural versus urban veterans' use of VA and non-VA inpatient addiction services. METHODS: By using a comprehensive data set of VA and non-VA hospitalizations for 294,748 VA enrollees who were residents of New York State from 1998 to 2000, this study examined admission rates for addiction treatment to VA and non-VA centers to determine how rates differed between rural veterans and urban veterans. RESULTS: Between 1998 and 2000 rural veterans obtained 67% of their inpatient addiction care from the VA, compared with 54% for urban veterans (p<.001). Compared with 1998 levels, the odds ratios of admission to VA facilities for inpatient detoxification fell for both rural and urban veterans to .80 in 1999 and .65 in 2000 (both p<.05). Although odds ratios of non-VA inpatient admission for addiction treatment were stable over time for urban veterans, those for rural veterans fell from 1998 values, falling to .76 in 1999 (not significant) and .62 in 2000 (p<.001) for detoxification and to .66 in 1999 (not significant) and .51 in 2000 for rehabilitation (p<.05). Odds ratios for urban veterans' admission to VA facilities for rehabilitation fell to .51 in terms of 1998 rates in 1999 and .38 in 2000, but rural veterans' odds ratios fell more, to .31 and .16, respectively (p<.001 for all). CONCLUSIONS: In New York regionalization of VA addiction services disproportionately affected rural veterans. Rural veterans experienced concurrent reductions in VA and non-VA inpatient addiction services. The VA and other health care policy makers should consider the potential unintended consequences to rural populations of resource reallocation. PMID- 17463349 TI - A longitudinal investigation of intimate partner violence among mothers with mental illness. AB - OBJECTIVE: Severe mental illness, substance use, and intimate partner violence have emerged as major intersecting public health problems that adversely and disproportionately impact the lives of women in the United States. This longitudinal study investigated the demographic and clinical correlates of intimate partner violence in a sample of 324 mothers with severe mental illness. METHODS: A secondary analysis of longitudinal data was conducted by using multiple logistic regression. Participants were part of a longitudinal, community based study of mothers with severe mental illness, which was aimed at understanding how these mothers viewed motherhood. The women were interviewed initially at baseline (interviews were conducted between 1995 and 1996) and then about 20 months later at follow-up (interviews were conducted between 1997 and 1998). RESULTS: At follow-up the prevalence rate of intimate partner violence was 19%. Multiple logistic regression analyses showed a significant positive relationship between alcohol and drug misuse at baseline and intimate partner violence at follow-up, indicating that women with a co-occurring diagnosis of a substance use disorder (dual diagnosis) were more likely than women without such a diagnosis to report intimate partner violence. The number of lifetime psychiatric hospitalizations and the number of symptoms related to psychiatric disability exhibited at baseline were positively associated with intimate partner violence at follow-up, and age was inversely associated with intimate partner violence. CONCLUSIONS: Mental health professionals serving mothers with mental health problems need to be aware of and prepared to assess the significant correlation between these intersecting public health problems in order to influence successful interventions. Particular attention must be given to the special treatment needs related to dual diagnosis and victimization and the impact of these factors on this vulnerable population. PMID- 17463350 TI - Differences in HIV care between patients with and without severe mental illness. AB - OBJECTIVE: This study explored how HIV care differs for infected persons with and without severe mental illness. METHODS: Data were obtained through interviews with and chart review of 295 patients with severe mental illness and HIV from public mental health agencies in Los Angeles County and New York City. Data were compared with data from 1,294 HIV patients without severe mental illness from a separate national probability sample. Measures were difficulty obtaining care, whether patients recommend their HIV care provider, hospital problem score, functional health status, and disability days. RESULTS: In Los Angeles, HIV patients with severe mental illness were more likely than those without severe mental illness to have difficulty obtaining care (p<.001); to not recommend their provider (10% versus 5%, p=.007); and to have problematic hospital care (p=.001), poor health status (p=.001), and more disability days (p<.001). In New York City, HIV patients with severe mental illness were more likely than patients without severe mental illness to have difficulty obtaining care (p=.002) and not recommend their provider (p=.02). The relationship between severe mental illness and health status in Los Angeles and access in New York City became insignificant after adjustment for sociodemographic factors, drug use, and CD4 cell count. Further adjustment for higher case management rates among HIV patients with severe mental illness reduced disparities only in the West. CONCLUSIONS: Patients with severe mental illness experienced more problems with HIV care than patients without severe mental illness, although high case management rates for patients with severe mental illness may have offset some problems. PMID- 17463351 TI - Social networks and their relationship to mental health service use and expenditures among Medicaid beneficiaries. AB - OBJECTIVE: This study examined the relationship between social networks and mental health services utilization and expenditures. METHODS: A sample of 522 Medicaid mental health consumers was randomly selected from the administrative records of Colorado's Department of Health Care Policy and Financing. The administrative records contain information on utilization of services and expenditures of Medicaid beneficiaries within Colorado's Mental Health Services. In addition to the administrative records, social network and psychosocial data were gathered through longitudinal survey interviews. The interviews were conducted at six-month intervals between 1994 and 1997. Measures used in the regression analysis included demographic characteristics, clinical diagnoses, the social network index, expenditures, and utilization variables. RESULTS: The social network index was positively associated with utilization of and expenditures for inpatient services in local hospitals but negatively associated with expenditures for inpatient services in state hospitals or outpatient services. Relationships with family were negatively related to expenditures for outpatient services. Relationships with friends were positively associated with utilization of and expenditures for psychiatric inpatient services in local hospitals. CONCLUSIONS: Consumers who had higher social network index scores utilized more inpatient psychiatric services in local hospitals and had higher expenditures than those who had lower scores. Consumers who had higher social network index scores also had lower expenditures for inpatient services in state hospitals and outpatient services than those who have lower scores. Findings suggest that social network is associated with mental health utilization and expenditures in various ways, associations that need to be researched further. PMID- 17463352 TI - Matching services to patients' beliefs about depression in Dublin, Ireland. AB - OBJECTIVES: This study explored tensions between biomedical and psychosocial illness models and between the public health goal of stigma reduction and patients' perceptions of helplessness. Patients' etiological beliefs and use of language are considered as possible pathways to improved care. METHODS: Qualitative and quantitative data were obtained from 18 adults attending a mood disorders support group in Dublin, Ireland. RESULTS: Respondents reported feelings of helplessness concerning the occurrence of depressive episodes. When describing episodes of wellness and depression, respondents used a "light switch" metaphor to indicate this lack of control. For several respondents, acceptance of biomedical causality was linked to helplessness. In contrast, spiritual beliefs offered hope. CONCLUSIONS: Alleviating patients' sense of responsibility may reduce their guilt, but patients' perceptions of illness uncontrollability may diminish their sense of agency, reinforce depressive attributional styles, and undermine overall treatment outcomes. Communication and the patients' involvement in decisions are vital to treatment. PMID- 17463353 TI - Clinical presentation and treatment outcome of African Americans with complicated grief. AB - OBJECTIVE: This study sought to examine whether ethnic differences occur in the presentation of patients with complicated grief or their treatment outcome. METHODS: Analyses of a randomized controlled trial comparing a novel psychotherapy for complicated grief with interpersonal psychotherapy contrasted the clinical presentation, treatment alliance, and rates of treatment completion and response for 19 African Americans with complicated grief and 19 Caucasian Americans with complicated grief matched by sex, age, and baseline grief severity. Participants were randomly assigned to receive 16 sessions of either standard interpersonal psychotherapy or interpersonal psychotherapy enhanced with focused complicated grief components. RESULTS: No differences were found in any clinical or treatment-related measure. CONCLUSIONS: African Americans and Caucasian Americans with complicated grief did not differ significantly in clinical presentation, treatment alliance, treatment completion, and outcome. The results suggest that standard treatment for complicated grief can be provided successfully for different racial and economic groups. PMID- 17463354 TI - Clinical characteristics and health service use of veterans with comorbid bipolar disorder and PTSD. AB - OBJECTIVE: This study aimed to increase the understanding of the clinical characteristics and utilization of health services among veterans with comorbid bipolar disorder and posttraumatic stress disorder (PTSD). METHODS: A retrospective chart review was conducted that examined the clinical and health service use data of 139 male veterans with bipolar disorder (N=49), PTSD (N=49), or comorbid bipolar disorder and PTSD (N=41) who obtained services over two years from the Department of Veterans Affairs Salt Lake City Health Care System. RESULTS: Compared with patients with bipolar disorder or those with PTSD alone, those with both conditions had significantly higher mean Clinical Global Impression-Severity scores and required more frequent inpatient psychiatric treatment. Patients with both conditions and those with bipolar disorder were significantly less likely to have received psychotherapy and antidepressant pharmacotherapy than patients with PTSD. CONCLUSIONS: Patients with both conditions have a more severe illness course than those with bipolar disorder alone. Psychotherapy and antidepressant pharmacotherapy may be underutilized among patients with comorbid bipolar disorder and PTSD. PMID- 17463355 TI - An update on the South Carolina trauma initiative. AB - Seven years ago the South Carolina Department of Mental Health developed a statewide effort to improve its response to consumers who are trauma survivors. The initiative's goals included sensitizing stakeholders to the impact of trauma, influencing policies, educating and training clinicians, addressing concerns about the safety and dignity of psychiatric settings (for example, sanctuary trauma), and increasing knowledge by supporting a strong empirical research platform. In the brief report presented here, the authors describe the past seven years in terms of obstacles encountered, progress, and future directions of this initiative in the hope of providing information and guidance to other state systems. PMID- 17463356 TI - Use of CBT to treat depression among patients on hemodialysis. PMID- 17463357 TI - CBT for psychosis for long-term inpatients with a forensic history. PMID- 17463358 TI - Let's get organized: an intervention for persons with co-occurring disorders. PMID- 17463360 TI - Consumer-directed behavioral health care. PMID- 17463361 TI - Proceedings of Reproductive Biomechanics Track of the 5th World Congress of Biomechanics, July 29-August 4, 2006, Munich, Germany. PMID- 17463362 TI - Sexuality, health, and human rights. PMID- 17463363 TI - The roles of abstinence and contraception in declining pregnancy rates. PMID- 17463364 TI - Recently arrived immigrant Latino men identify community approaches to promote HIV prevention. PMID- 17463365 TI - Male-to-female transgender and transsexual clients of HIV service programs in Los Angeles County, California. AB - Data on HIV risk were collected with the Countywide Risk Assessment Survey from 2126 participants; 92 were male-to-female transgender persons (i.e., cross dressers, and those who identify with the opposite sex), and 9 were male-to female transsexual individuals (i.e., those who have undergone gender reassignment surgery or other procedures). Transgender-identified individuals were more likely than the rest of the sample to have received hormone injections, offensive comments, and HIV testing; injected hormones with a used needle; been Asian or American Indian; been paid for sexual intercourse; and lived in unstable housing but less likely to have used heroin. Transgender-identified individuals are at high risk for HIV infection because of reuse of needles and being paid for sexual intercourse. PMID- 17463366 TI - Modern marriage, men's extramarital sex, and HIV risk in southeastern Nigeria. AB - For women in Nigeria, as in many settings, simply being married can contribute to the risk of contracting HIV. I studied men's extramarital sexual behavior in the context of modern marriage in southeastern Nigeria. The results indicate that the social organization of infidelity is shaped by economic inequality, aspirations for modern lifestyles, gender disparities, and contradictory moralities. It is men's anxieties and ambivalence about masculinity, sexual morality, and social reputation in the context of seeking modern lifestyles--rather than immoral sexual behavior and traditional culture--that exacerbate the risks of HIV/AIDS. PMID- 17463367 TI - Men's extramarital sexuality in rural Papua New Guinea. AB - Married women in rural Papua New Guinea are at risk for HIV primarily because of their husbands' extramarital relationships. Labor migration puts these men in social contexts that encourage infidelity. Moreover, many men do not view sexual fidelity as necessary for achieving a happy marriage, but they view drinking and "looking for women" as important for male friendships. Although fear of HIV infection is increasing, the concern that men most often articulated about the consequences of extramarital infidelity was possible violent retaliation for "stealing" another man's wife. Therefore, divorced or separated women who exchange sex for money are considered to be "safe" partners. Interventions that promote fidelity will fail in the absence of a social and economic infrastructure that supports fidelity. PMID- 17463368 TI - The inevitability of infidelity: sexual reputation, social geographies, and marital HIV risk in rural Mexico. AB - Marriage presents the single greatest risk for HIV infection among women in rural Mexico. We drew on 6 months of participant observation, 20 marital case studies, 37 key informant interviews, and archival research to explore the factors that shape HIV risk among married women in one of the country's rural communities. We found that culturally constructed notions of reputation in this community lead to sexual behavior designed to minimize men's social risk (threats to one's social status or relationships), rather than viral risk and that men's desire for companionate intimacy may actually increase women's risk for HIV infection. We also describe the intertwining of reputation-based sexual identities with structurally patterned sexual geographies (i.e. the social spaces that shape sexual behavior). We propose that, because of the structural nature of men's extramarital sexual behavior, intervention development should concentrate on sexual geographies and risky spaces rather than risky behaviors or identities. PMID- 17463369 TI - Overweight and obesity in sexual-minority women: evidence from population-based data. AB - OBJECTIVE: We sought to determine whether lesbians have higher rates of overweight and obesity than women of other sexual orientations. METHODS: We compared population estimates of overweight and obesity across sexual orientation groups, using data from the 2002 National Survey of Family Growth. RESULTS: Adjusted multinomial logistic regression analyses showed lesbians have more than twice the odds of overweight (odds ratio [OR]=2.69; 95% confidence interval [CI]=1.40, 5.18) and obesity (OR=2.47; 95% CI=1.19, 5.09) as heterosexual women. Bisexuals and women who reported their sexual orientation as "something else" (besides heterosexual, lesbian, or bisexual) showed no such increase in the odds of overweight and obesity. CONCLUSIONS: Lesbian women have a higher prevalence of overweight and obesity than all other female sexual orientation groups. This finding suggests that lesbians are at greater risk for morbidity and mortality linked to overweight and obesity. This finding also highlights the need for interventions within this population. PMID- 17463370 TI - Racial/ethnic and age disparities in HIV prevalence and disease progression among men who have sex with men in the United States. AB - OBJECTIVES: We examined HIV diagnosis rates and disease progression among men who have sex with men (MSM) according to race/ethnicity and age. METHODS: Using data obtained from the national HIV/AIDS surveillance system, we examined trends in HIV diagnosis rates for 2001 through 2004 using Poisson regression. We used a standardized Kaplan-Meier method to determine differences in time of progression from HIV to AIDS and AIDS survival. RESULTS: HIV diagnosis rates were higher for Black and Hispanic than for White MSM, but trends within age groups from 2001 to 2004 did not differ by race/ethnicity. Diagnosis rates increased among MSM aged 13 to 19 years (14% per year), 20 to 24 years (13%), 25 to 29 years, and 40 to 54 years (3%-6%; P< or = .01 for each). The percentage of MSM who did not have AIDS 3 years after HIV diagnosis was lower among Black (66.8%; 95% confidence interval [CI]=66.1, 67.4) and Hispanic (68.1%; 95% CI=67.5, 68.8) than among White MSM (74.7%; 95% CI=74.2, 75.1). Three-year survival after AIDS diagnosis was lower for Black than for White or Hispanic MSM. CONCLUSIONS: HIV prevention efforts should target young and middle-aged MSM and must offer early diagnosis and treatment for all MSM. PMID- 17463371 TI - Physical health complaints among lesbians, gay men, and bisexual and homosexually experienced heterosexual individuals: results from the California Quality of Life Survey. AB - OBJECTIVES: We examined evidence that minority sexual orientation is associated with more-frequent reports of physical health complaints. We also investigated the possible role of HIV infection among gay men and higher rates of psychological distress among lesbians, gay men, and bisexually and homosexually experienced heterosexual individuals in generating these health disparities. METHODS: We used data from the California Quality of Life Survey (N=2272 adults) to examine associations between sexual orientation and self-reports about physical health status, common health conditions, disabilities, and psychological distress. RESULTS: Prevalent HIV infection was reported by nearly 18% of gay, bisexual, and homosexually experienced heterosexual men. Gay men and bisexual and homosexually experienced heterosexual individuals had higher levels of psychological distress compared with exclusively heterosexual individuals. Self reported physical health status varied by gender and by sexual orientation. CONCLUSIONS: Lesbians and bisexual and homosexually experienced heterosexual women reported a greater variety of health conditions and limitations compared with exclusively heterosexual women; however, these differences mostly disappeared when distress levels were taken into account. Among men, differences in health complaints appeared to reflect the ongoing burden of HIV and other sexually transmitted diseases in the gay male community. PMID- 17463372 TI - Women who report having sex with women: British national probability data on prevalence, sexual behaviors, and health outcomes. AB - OBJECTIVES: We estimated the prevalence of same-sex experience among women and compared women reporting sex with women and men and women reporting sex exclusively with women with women reporting sex exclusively with men, in terms of sociodemographics and sexual, reproductive, and general health risk behaviors and outcomes. METHODS: We used a British probability survey (n=6399 women, aged 16 to 44 years) conducted from 1999 to 2001 with face-to-face interviewing and computer assisted self-interviewing. RESULTS: We found that 4.9% of the women reported same-sex partner(s) ever; 2.8% reported sex with women in the past 5 years (n=178); 85.0% of these women also reported male partner(s) in this time. Compared with women who reported sex exclusively with men, women who reported sex with women and men reported significantly greater male partner numbers, unsafe sex, smoking, alcohol consumption, and intravenous drug use and had an increased likelihood of induced abortion and sexually transmitted infection diagnoses (age adjusted odds ratios=3.07 and 4.41, respectively). CONCLUSIONS: For women, a history of sex with women may be a marker for increased risk of adverse sexual, reproductive, and general health outcomes compared with women who reported sex exclusively with men. A nonjudgmental review of female patients' sexual history should help practitioners discuss risks that women may face. PMID- 17463373 TI - Research, curricula, and resources related to lesbian, gay, bisexual, and transgender health in US schools of public health. AB - To assess the extent to which public health schools conduct research, offer planned curricula, and provide resources related to lesbian, gay, bisexual, and transgender health, we mailed a self-administered questionnaire to individual department chairpersons at each school. Survey results suggested that departmental lesbian, gay, bisexual, and transgender research and curricular activities extending beyond HIV and AIDS were uncommon in most public health school programs. Expanding lesbian, gay, bisexual, and transgender health research and curricula may help health professionals improve their response to lesbian, gay, bisexual, and transgender health disparities. PMID- 17463374 TI - The effect of social support on mental and behavioral outcomes among adolescents with parents with HIV/AIDS. AB - OBJECTIVES: We examined the associations between social support and mental and behavioral outcomes among adolescents whose parents were infected with or died of HIV/AIDS. METHODS: Families (parents who were HIV infected and their adolescent children) were randomly assigned to a coping skills intervention or a standard care group. After completing the intervention, the parents and adolescents were assessed for 2 years. RESULTS: Adolescents who had more social support providers reported significantly lower levels of depression and fewer conduct problems; adolescents who had more negative influence from role models reported more behavior problems. Reductions in depression, multiple problem behaviors, and conduct problems were significantly associated with better social support. CONCLUSIONS: Our findings underscore the complex relations between social support and mental and behavioral outcomes among adolescents affected by HIV/AIDS. Future prevention programs must focus on increasing social support to reduce negative outcomes among adolescents affected by HIV/AIDS as well as the need to reduce influence from negative role models. PMID- 17463375 TI - Directly observed highly active antiretroviral therapy for HIV-infected children in Cambodia. AB - Antiretroviral medications are becoming available for HIV-infected children in resource-limited settings. Maryknoll, an international Catholic charity, provided directly observed antiretroviral therapy to HIV-infected children in Phnom Penh, Cambodia. Child care workers administered generic antiretroviral drugs twice daily to children, ensuring adherence. Treatment began with 117 late-stage HIV infected children; 22 died of AIDS during the first 6 months. The rest were treated for at least 6 months and showed CD4 count increases comparable to those achieved in US and European children. Staffing cost for this program was approximately US $5 per child per month, or 15% more than the price of the medications. Drug toxicities were uncommon and easily managed. Directly observed antiretroviral therapy appears to be a promising, low-cost strategy for ensuring adherent treatment for HIV-infected children in a resource-limited setting. PMID- 17463376 TI - Bisexual Latino men and HIV and sexually transmitted infections risk: an exploratory analysis. AB - OBJECTIVES: We sought to determine whether there were differences in sexual risk among behaviorally and self-identified bisexual men, men who reported having sex with both men and women without reporting a bisexual identity and men who self identified as bisexual but reported only recent homosexual behavior over the past 6 months. METHODS: secondary data analysis, we conducted stepwise linear regression equation modeling to determine which factors were significant predictors of sexual risk among various subgroups of bisexual Latino men. RESULTS: Having sex with women, regardless of sexual identity, increased the likelihood of insertive unprotected anal intercourse with men. Bisexual identity was not statistically associated with unprotected anal intercourse with men. CONCLUSIONS: Future studies should begin to examine differences within groups rather than focusing on group comparisons that yield limited insights into sexual risk predictors for homosexually and bisexually active men. Further research that explores risk and protective factors in the sexual lives of Latino bisexual men is also needed. PMID- 17463377 TI - "He won't use condoms": HIV-infected women's struggles in primary relationships with serodiscordant partners. AB - We investigated the sexual behaviors of 55 HIV-infected women in Wisconsin who narrated their lives in 10 interviews over 2 years during 2000 to 2003. We sought to examine the interpersonal situations in which sexual risk occurred. During the prospective period, 58% (32) were abstinent and 24% (13) practiced safe sex exclusively. The remaining 18% (10) engaged in unprotected sexual intercourse, but only in primary partnerships, almost all of which were with serodiscordant partners. We focused on experiential detail and narrative depth of 10 women who had sex without condoms. These narratives demonstrate how the women attempted to initiate condom use but engaged in unprotected sexual intercourse regularly at the insistence of their partners. Consequently, these women lived in trepidation of causing their partners' sickness and death. PMID- 17463378 TI - Tip of the Iceberg: young men who have sex with men, the Internet, and HIV risk. AB - OBJECTIVES: We examined the prevalence of Internet use for meeting sexual partners (Internet partners) and HIV risk behaviors associated with this use among young men who have sex with men (aged 16-24 years). METHODS: A sample of 270 young men who have sex with men completed a computer-assisted survey. We used bivariate chi(2) analyses and hierarchical logistic regression to assess factors associated with Internet-facilitated sexual encounters. RESULTS: Using the Internet to meet sexual partners was common; 48% of our sample had sexual relations with a partner they met online. Of these, only 53% used condoms consistently, and 47% reported having sexual partners older (>4 years) than themselves. Regression analyses showed increased age, White race/ethnicity, history of unprotected anal intercourse, multiple anal intercourse partners, and engaging in sexual activity at a sex club or a bathhouse were associated with meeting sexual partners through the Internet. Only history of unprotected anal intercourse was associated with risky sexual behaviors with Internet partners (P<0.025). CONCLUSIONS: Young men who have sex with men and who seek partners online also engage in other behaviors that place them at risk for HIV and other sexually transmitted infections. PMID- 17463379 TI - Growth trajectories of sexual risk behavior in adolescence and young adulthood. AB - OBJECTIVES: Adolescence and young adulthood (ages 18-25 years) are periods of development and change, which include experimentation with and adoption of new roles and behaviors. We investigated longitudinal trajectories of sexual risk behaviors across these time periods and how these trajectories may be different for varying demographic groups. METHODS: We developed multilevel growth models of sexual risk behavior for a predominantly African American sample (n=847) that was followed for 8 years, from adolescence to young adulthood. We investigated differences in growth parameters by race/ethnicity and gender and their interactions. RESULTS: The final model included linear and quadratic terms for both adolescence and young adulthood, indicating acceleration of sexual risk behaviors during adolescence and a peak and deceleration during young adulthood. African American males exhibited the highest rate of sexual risk behavior in ninth grade, yet had the slowest rate of growth. Compared with their White peers, African American males and females exhibited less sexual risk behavior during young adulthood. CONCLUSIONS: Our results suggest that youths of different races/ethnicities and genders exhibit varying sexual risk behavior trajectories. PMID- 17463380 TI - Mycoplasma genitalium among young adults in the United States: an emerging sexually transmitted infection. AB - OBJECTIVES: We sought to determine the prevalence of and risk factors associated with Mycoplasma genitalium infection in a nationally representative sample of young adults in the United States. METHODS: Urine specimens from 1714 women and 1218 men who participated in Wave III of the National Longitudinal Study of Adolescent Health (N=14322) were tested for M genitalium. Poststratification sampling weights were used to generate nationally representative estimates. RESULTS: The prevalence of M genitalium was 1.0% compared with 0.4%, 4.2%, and 2.3% for gonococcal, chlamydial, and trichomonal infections, respectively. No M genitalium-positive individuals reported symptoms of discharge. M genitalium prevalence among those who reported vaginal intercourse was 1.1% compared with 0.05% among those who did not. In multivariate analyses, M genitalium prevalence was 11 times higher among respondents who reported living with a sexual partner, 7 times higher among Blacks, and 4 times higher among those who used condoms during their last vaginal intercourse. Prevalence of M genitalium increased by 10% for each additional sexual partner. CONCLUSIONS: M genitalium was more prevalent than Neisseria gonorrhoeae but less prevalent than Chlamydia trachomatis, and it was strongly associated with sexual activity. PMID- 17463381 TI - Ethics in public health research: minding the gaps: a reassessment of the challenges to safe motherhood. AB - Maternal and perinatal mortality reduction has remained a priority on the international health agenda for nearly 2 decades. During this time, strategies for achieving these goals have shifted in emphasis from prevention of pregnancies to provision of care. Robust evidence is limited, particularly regarding what works best in delivering care in specific health system settings and at the population level. We describe the limited evidence base using a framework that highlights the consequences of the major gaps in measurement, evidence, and action, and we discuss existing opportunities for bridging these gaps at the policy level. Capitalizing on current global policy interests and generating demand-driven evidence is a priority for enabling documentation of progress toward reaching the United Nations Millennium Development Goals for 2015. PMID- 17463382 TI - REAL men: a group-randomized trial of an HIV prevention intervention for adolescent boys. AB - OBJECTIVES: We tested the efficacy of an intervention among 11- to 14-year-old adolescent boys to promote delay of sexual intercourse, condom use among those who were sexually active, and communication on sexuality between fathers (or father figures) and sons. METHODS: Sites were randomly assigned to the intervention and control groups. Assessments were conducted prior to the intervention and at 3-, 6-, and 12-month follow-up interviews. RESULTS: A total of 277 fathers and their sons completed baseline assessments. Most participants were African American, and most fathers lived with their sons. Significantly higher rates of sexual abstinence and condom use and of intent to delay initiation of sexual intercourse were observed among adolescent boys whose fathers participated in the intervention. Fathers in the intervention group reported significantly more discussions about sexuality and greater intentions to discuss sexuality than did control-group fathers. CONCLUSIONS: The study demonstrates that fathers can serve as an important educator on HIV prevention and sexuality for their sons. PMID- 17463383 TI - The effectiveness of state and national policy on the implementation of perinatal HIV prevention interventions. AB - OBJECTIVES: The 1994 and 1995 US Public Health Service Guidelines regarding HIV testing and treatment for pregnant women and the resulting 1995 California law mandating an HIV test and treatment offer to every pregnant woman aim to reduce perinatal HIV transmission. However, the effectiveness of such policies after implementation is often unclear. We analyzed the association between these policies and offers of HIV tests and treatment to HIV-infected women in California. METHODS: Data from active, population-based surveillance of 496 HIV infected women and their infants, collected from 1987 to 2002, were analyzed to compare rates of offers of HIV tests and treatment before and after 1996. RESULTS: We found significant increases in offers of HIV tests (P<.001) and offers of treatment (P<.001) when we compared women who delivered between 1987 and 1995 with those who delivered between 1996 and 2002. Receipt of prenatal care was the major predictor of both test and treatment offer. A significant shift in reported HIV risk factors was also evident between the 2 groups. CONCLUSIONS: Our findings of increased offers of HIV tests and treatment to HIV-infected pregnant women suggest that the national guidelines and the 1996 California law improved health care for these women, which may lessen the risk of perinatal HIV transmission. PMID- 17463384 TI - Prevalence of HIV infection and predictors of high-transmission sexual risk behaviors among men who have sex with men. AB - OBJECTIVES: We sought to determine the prevalence of HIV and novel cofactors of high-transmission-risk behavior in a probability sample of men who have sex with men (MSM). METHODS: We performed a cross-sectional telephone survey of 1976 adult MSM in San Francisco. RESULTS: We found an HIV prevalence of 25.2%. Predictors of unprotected insertive anal intercourse with a serodiscordant (not having the same HIV/AIDS serostatus) partner among HIV-infected men included use of Viagra and a greater number of partners in the past 12 months. Unprotected receptive anal intercourse with a serodiscordant partner among men not known to be HIV infected was independently associated with having lived in San Francisco for less than 1 year, use of crystal methamphetamine and amyl nitrites, a greater number of partners, and agreement with the statement, "You are less careful about being safe with sex or drugs than you were several years ago because there are better treatments for HIV now." CONCLUSIONS: Strategies to prevent HIV for urban MSM should focus on new predictors of HIV transmission. PMID- 17463385 TI - Reproductive health of adolescent girls perinatally infected with HIV. AB - OBJECTIVES: We sought to describe the reproductive health of adolescent girls perinatally infected with HIV. METHODS: We estimated the incidence of first pregnancy, genital infections, and abnormal cervical cytology for 638 girls aged 13 years and older in the Pediatric AIDS Clinical Trials Group protocol 219C. RESULTS: Thirty-eight girls became pregnant, for a first pregnancy rate of 18.8/1000 person-years; 7 of these girls had additional pregnancies (95% confidence interval [CI]=13.3, 25.7). Thirty-two pregnancies resulted in live births. All girls received antiretroviral therapy during pregnancy. One infant was HIV infected, 29 were uninfected, and 2 had unknown infection status, for a rate of mother-to-child transmission of HIV in infants with known infection status of 3.3% (95% CI=0.1, 18.6). Condylomata and trichomoniasis were the most frequent genital infections. Forty-eight (47.5%) of 101 girls with Papanicolaou test examinations had abnormal cervical cytology, including atypical cells of undetermined significance (n=18), low-grade squamous intraepithelial lesions (SIL; n=27), and high-grade SIL (n=3). Many abnormalities persisted despite intervention. CONCLUSIONS: Pregnancy rates were lower and cervical abnormalities were higher than among non-HIV-infected adolescents. These findings underscore the importance of Papanicolaou tests and promotion of safer sexual practices in this population. PMID- 17463386 TI - History of childhood sexual abuse and HIV risk behaviors in homosexual and bisexual men. AB - OBJECTIVES: We examined the prevalence and frequency of childhood sexual abuse and their association with sexual risk among a sample of gay and bisexual men. Methods. Cross-sectional data were collected by survey from randomly selected gay and bisexual men who attended the 1997 and 1998 Minneapolis/St. Paul Gay, Lesbian, Bisexual, and Transgender Pride Festivals. Data included demographics, sexual activity, history of childhood sexual abuse, HIV status, history of sexually transmitted infection, use of sex-related drugs (such as crack, cocaine, Ecstasy, amyl nitrate, crystal methamphetamine, and Special K), and history of exchanging sex for payment. Results. childhood sexual abuse was reported by 15.5% of the survey respondents (n = 134). Those who reported experiencing abuse regularly were more likely to (1) be HIV positive, (2) have exchanged sex for payment, and (3) be a current user of sex-related drugs. Neither unsafe sex nor sexually transmitted infections were associated with childhood sexual abuse. CONCLUSIONS: These findings show that more than 1 in 7 gay and bisexual men in a non-clinical, festival-based setting were victims of childhood sexual abuse and that childhood sexual abuse was associated with alarmingly high rates of men who were HIV infected and antecedent risk behaviors. PMID- 17463387 TI - Trends in primary and secondary syphilis among men who have sex with men in the United States. AB - OBJECTIVES: We assessed the epidemiology of primary and secondary syphilis in the United States and estimated the percentages of cases occurring among men who have sex with men (MSM). METHODS: We reviewed US syphilis surveillance data from 1990 through 2003. We estimated the number of cases occurring among MSM by modeling changes in the ratio of syphilis cases among men to cases among women. RESULTS: During 1990 through 2000, the rate of primary and secondary syphilis decreased 90% overall, declining 90% among men and 89% among women. The overall rate increased 19% between 2000 and 2003, reflecting a 62% increase among men and a 53% decrease among women. In 2003, an estimated 62% of reported cases occurred among MSM. CONCLUSIONS: Increasing syphilis cases among MSM account for most of the recent overall increase in rates and may be a harbinger of increasing rates of HIV infection among MSM. National efforts are under way to improve monitoring of syphilis trends, better understand factors associated with the observed increases, and improve efforts to prevent syphilis transmission. PMID- 17463388 TI - Association between condom use at sexual debut and subsequent sexual trajectories: a longitudinal study using biomarkers. AB - OBJECTIVES: We compared subsequent sexual behaviors and risk of sexually transmitted infections among adolescents who did and did not use a condom at their sexual debut. METHODS: We derived data from the National Longitudinal Study of Adolescent Health, which followed a sample of 4018 sexually active adolescents between 1994 and 2002. During waves I, II, and III of the study, data on sexual behavior were gathered, and at wave III urine specimens were collected to test for sexually transmitted infections. RESULTS: Among interviewed adolescents, those who reported condom use at their debut were more likely than those who did not use condoms at their debut to report condom use at their most recent intercourse (on average 6.8 years after sexual debut), and they were only half as likely to test positive for chlamydia or gonorrhea (adjusted odds ratio=0.50; 95% confidence interval=0.26, 0.95). Reported lifetime numbers of sexual partners did not differ between the 2 groups. CONCLUSIONS: Adolescents who use condoms at their sexual debut do not report more sexual partners, are more likely to engage in subsequent protective behaviors, and experience fewer sexually transmitted infections than do adolescents who do not use condoms at their sexual debut. PMID- 17463389 TI - Managing Medicare's HIV caseload in the era of suppressive therapy. AB - OBJECTIVES: The 1996 introduction of antiretroviral medications changed Medicare's role in providing HIV care. We analyzed Medicare's patient database in an effort to document the new HIV therapies' effects on expenditures and outcomes. METHODS: We examined the medical billing records of a 5% national Medicare sample from 1997 through 2003. The cohort was stratified by year and categorized by age, race/ethnicity, gender, and Medicare status. Population summaries were categorized according to presence of major chronic diseases and HIV-related conditions. RESULTS: The number of Medicare beneficiaries with HIV increased from 42520 in 1997 to 76500 in 2003, whereas mortality among this group fell by 35%. HIV-associated infections declined by as much as 43% (mycoses). Heart and liver disease and diabetes increased by more than 50%. Adjusted annual per person Medicare expenditures fell 28%; expenditures were 49% higher for Blacks than for Whites. CONCLUSIONS: Improved HIV medical management has led to fewer deaths and has shifted treatment toward chronic care. However, successful management is complicated by conditions that have not been historically linked to HIV and whose effects vary according to race/ethnicity. PMID- 17463390 TI - Changes in prevalence of HIV infection and sexual risk behavior in men who have sex with men in San Francisco: 1997 2002. AB - OBJECTIVES: We assessed differences in HIV prevalence and sexual risk behavior among men who have sex with men (MSM) between 1997 and 2002 in San Francisco. METHODS: We used 2 population-based random-digit-dial telephone surveys of MSM households in San Francisco in 1997 (n=915) and 2002 (n=879). RESULTS: Estimated HIV prevalence increased from 19.6% in 1997 to 26.8% in 2002. Measures of sexual risk also increased. Unprotected anal intercourse with a partner of different or unknown HIV serostatus increased from 9.3% to 14.6%. Mean number of male partners increased from 10.7 to 13.8. The largest reported increase was 18.9% to 26.8% for "serosorting," or choosing unprotected anal intercourse partners believed to have the same HIV serostatus as oneself. Men aged 30 to 50 reported the largest increase in unprotected anal intercourse, whereas men aged 18 to 29 reported the largest increase in serosorting. Changes in the age distribution did not explain the increase in risky behavior. CONCLUSIONS: Both HIV prevalence and sexual risk increased substantially among MSM in San Francisco between 1997 and 2002. Serosorting is being adopted more frequently than condom use by young MSM, but its effectiveness as a harm reduction strategy is not known. PMID- 17463391 TI - Effects on sexual risk behavior and STD rate of brief HIV/STD prevention interventions for African American women in primary care settings. AB - OBJECTIVES: We tested the efficacy of brief HIV/sexually transmitted disease (STD) risk-reduction interventions for African American women in primary care settings. METHODS: In a randomized controlled trial, 564 African American women recruited at a Newark, NJ, inner-city women's health clinic were assigned to a 20 minute one-on-one HIV/STD behavioral skill-building intervention, 200-minute group HIV/STD behavioral skill-building intervention, 20-minute one-on-one HIV/STD information intervention, 200-minute group HIV/STD information intervention, or 200-minute health intervention control group. Primary outcomes were self-reported sexual behaviors in the previous 3 months; secondary outcome was STD incidence. RESULTS: At 12-month follow-up, participants in the skill building interventions reported less unprotected sexual intercourse than did participants in the information interventions (Cohen's d [d]=0.23, P=.02), reported a greater proportion of protected sexual intercourse than did information intervention participants (d=0.21, P=.05) and control participants (d=0.24, P=.03), and were less likely to test positive for an STD than were control participants (d=0.20, P=.03). CONCLUSIONS: This study suggests that brief single-session, one-on-one or group skill-building interventions may reduce HIV/STD risk behaviors and STD morbidity among inner-city African American women in primary care settings. PMID- 17463392 TI - Airway hyperresponsiveness, remodeling, and smooth muscle mass: right answer, wrong reason? AB - We quantified the effects of airway wall remodeling upon airway smooth muscle (ASM) shortening. Isolated ASM from sheep was attached to a servo-controller that applied a physiologic load. This load could be altered to reflect specified changes of airway wall geometry, elasticity, parenchymal tethering, transpulmonary pressure (P(L)), and fluctuations in P(L) associated with breathing. Starting at a reference length (L(ref)), ASM was stimulated with acetlycholine and held at constant P(L) of 4 cm H(2)O for 2 h. When all compartments were thickened to simulate the asthmatic airway but P(L) was held fixed, ASM shortened much more than that in the normal airway (to 0.52 L(ref) versus 0.66 L(ref)). When breathing with deep inspirations (DIs) was initiated, within the first three DIs the ASM in the normal airway lengthened to 0.84 L(ref), whereas that in the asthmatic airway remained stuck at 0.53 L(ref). Thickening of the smooth muscle layer alone produced the greatest muscle shortening (to 0.47 L(ref)) when compared with thickening of only submucosal (to 0.67 L(ref)) or only adventitial (to 0.62 L(ref)) compartments. With increased ASM mass, the ASM failed to lengthen in response to DIs, whereas in the airway with thickened submucosal and adventitial layers ASM lengthened dramatically (to 0.83 L(ref)). These findings confirm the long-held conclusion that increased muscle mass is the functionally dominant derangement, but mechanisms accounting for this conclusion differ dramatically from those previously presumed. Furthermore, increased ASM mass explained both hyperresponsiveness and the failure of a DI to relax the asthmatic airway. PMID- 17463393 TI - A potentiating effect of endogenous NO in the physiologic secretion from airway submucosal glands. AB - It is known that several second messengers, such as Ca(2+) or cAMP, play important roles in the intracellular pathway of electrolyte secretion in tracheal submucosal gland. However, the participation of cGMP, and therefore nitric oxide (NO), is not well understood. To investigate the physiologic role of NO, we first examined whether tracheal glands can synthesize NO in response to acetylcholine (ACh), and then whether endogenous NO has some effects on the ACh-triggered ionic currents. From the experiments using the NO-specific fluorescent indicator 4,5 diaminofluorescein diacetate salt (DAF-2DA), we found that a physiologically relevant low dose of ACh (100 nM) stimulated the endogenous NO synthesis, and it was almost completely suppressed in the presence of the nonspecific NO synthase (NOS) inhibitor Nomega-Nitro-L-arginine Methyl Ester Hydrochloride (L-NAME) or the neuronal NOS (nNOS)-specific inhibitor 7-Nitroindazole (7-NI). Patch-clamp experiments revealed that both the NOS inhibitors (L-NAME or 7-NI) and cGK inhibitors (KT-5823 or Rp-8-Br-cGMP) partially decreased ionic currents induced by 30 nM of ACh, but not in the case of 300 nM of ACh. Our results indicate that NO can be synthesized through the activation of nNOS endogenously and has potentiating effects on the gland secretion, under a physiologically relevant ACh stimulation. When cells were stimulated by an inadequately potent dose of ACh, which caused an excess elevation in [Ca(2+)](i), the cells were desensitized. Therefore, due to NO, gland cells become more sensitive to calcium signaling and are able to maintain electrolyte secretion without desensitization. PMID- 17463394 TI - Role of the SDF-1/CXCR4 axis in the pathogenesis of lung injury and fibrosis. AB - Stromal cell-derived factor-1 (SDF-1) participates in mobilizing bone marrow derived stem cells, via its receptor CXCR4. We studied the role of the SDF 1/CXCR4 axis in a rodent model of bleomycin-induced lung injury in C57BL/6 wild type and matrix metalloproteinase (MMP)-9 knockout mice. After intratracheal instillation of bleomycin, SDF-1 levels in serum and bronchial alveolar lavage fluid increased. These changes were accompanied by increased numbers of CXCR4(+) cells in the lung and a decrease in a population of CXCR4(+) cells in the bone marrow that did not occur in MMP-9(-)/(-) mice. Both SDF-1 and lung lysates from bleomycin-treated mice induced migration of bone marrow-derived stem cells in vitro that was blocked by a CXCR4 antagonist, TN14003. Treatment of mice with TN14003 with bleomycin-induced lung injury significantly attenuated lung fibrosis. Lung tissue from patients with idiopathic pulmonary fibrosis had higher numbers of cells expressing both SDF-1 and CXCR4 than did normal lungs. Our data suggest that the SDF-1/CXCR4 axis is important in the complex sequence of events triggered by bleomycin exposure that eventuates in lung repair. SDF-1 participates in mobilizing bone marrow-derived stem cells, via its receptor CXCR4. PMID- 17463396 TI - Gonadotropins and ovarian cancer. AB - Ovarian epithelial cancer (OEC) accounts for 90% of all ovarian cancers and is the leading cause of death from gynecological cancers in North America and Europe. Despite its clinical significance, the factors that regulate the development and progression of ovarian cancer are among the least understood of all major human malignancies. The two gonadotropins, FSH and LH, are key regulators of ovarian cell functions, and the potential role of gonadotropins in the pathogenesis of ovarian cancer is suggested. Ovarian carcinomas have been found to express specific receptors for gonadotropins. The presence of gonadotropins in ovarian tumor fluid suggests the importance of these factors in the transformation and progression of ovarian cancers as well as being prognostic indicators. Functionally, there is evidence showing a direct action of gonadotropins on ovarian tumor cell growth. This review summarizes the key findings and recent advances in our understanding of these peptide hormones in ovarian cancer development and progression and their role in potential future cancer therapy. We will first discuss the supporting evidence and controversies in the "gonadotropin theory" and the use of animal models for exploring the involvement of gonadotropins in the etiology of ovarian cancer. The role of gonadotropins in regulating the proliferation, survival, and metastasis of OEC is next summarized. Relevant data from ovarian surface epithelium, which is widely believed to be the precursor of OEC, are also described. Finally, we will discuss the clinical applications of gonadotropins in ovarian cancer and the recent progress in drug development. PMID- 17463395 TI - Central role of Muc5ac expression in mucous metaplasia and its regulation by conserved 5' elements. AB - Mucus hypersecretion contributes to morbidity and mortality in many obstructive lung diseases. Gel-forming mucins are the chief glycoprotein components of airway mucus, and elevated expression of these during mucous metaplasia precedes the hypersecretory phenotype. Five orthologous genes (MUC2, MUC5AC, MUC5B, MUC6, and MUC19) encode the mammalian gel-forming mucin family, and several have been implicated in asthma, cystic fibrosis, and chronic obstructive pulmonary disease pathologies. However, in the absence of a comprehensive analysis, their relative contributions remain unclear. Here, we assess the expression of the entire gel forming mucin gene family in allergic mouse airways and show that Muc5ac is the predominant gel-forming mucin induced. We previously showed that the induction of mucous metaplasia in ovalbumin-sensitized and -challenged mouse lungs occurs within bronchial Clara cells. The temporal induction and localization of Muc5ac transcripts correlate with the induced expression and localization of mucin glycoproteins in bronchial airways. To better understand the tight regulation of Muc5ac expression, we analyzed all available 5'-flanking sequences of mammalian MUC5AC orthologs and identified evolutionarily conserved regions within domains proximal to the mRNA coding region. Analysis of luciferase reporter gene activity in a mouse transformed Clara cell line demonstrates that this region possesses strong promoter activity and harbors multiple conserved transcription factor binding motifs. In particular, SMAD4 and HIF-1alpha bind to the promoter, and mutation of their recognition motifs abolishes promoter function. In conclusion, Muc5ac expression is the central event in antigen-induced mucous metaplasia, and phylogenetically conserved 5' noncoding domains control its regulation. PMID- 17463404 TI - The Medicare approach: political choice and American institutions. AB - The most significant health reform in American history was the passage of Medicare in 1965, but this was an accomplishment born of defeat. Medicare was designed and understood by its early promoters as an approach to health reform, not simply as a discrete program for a distinct target population. Although Medicare incrementalism has tended to be shunted aside when the opportunities for health reform are most promising, the final years of the Johnson administration reveal previously underappreciated efforts to expand Medicare eligibility to large new population groups and offer insights into the continuing potential of Medicare incrementalism in our own time. PMID- 17463405 TI - Through the looking glass: the politics of the Medicare Prescription Drug, Improvement, and Modernization Act. AB - The 2003 Medicare Prescription Drug, Improvement, and Modernization Act (MMA) represents a watershed event in Medicare politics. This essay explores the political lessons and policy legacies of the MMA, focusing on how the MMA fits with and deviates from established patterns in Medicare politics and how it will affect future reform. PMID- 17463406 TI - Protecting Medicare: the best defense is a good offense. AB - Traditional Medicare is being threatened from two political directions. The current Republican coalition, on the right, simply dislikes social insurance in principle. It seeks privatization for its own sake. Another perspective, centrist and well established among political and economic elites, worries that the program is "unaffordable," whatever its basic merits. Defenders of traditional Medicare need to address both threats by explaining why the budgetary fears are misconceived and why privatization is simply a bad idea. In order to do this, they need to take the budgetary high ground, argue more strenuously for short term cost controls, and criticize the extra spending that the Bush administration has used to encourage private plans within Medicare. Defenders of social insurance should also seek good policy and political allies by proposing that Medicare's network of providers, prices, and administration be made available to employers (and other pools) in much the way that self-insured groups currently rent networks from private insurers. PMID- 17463407 TI - Secret weapon: the "new" Medicare as a route to health security. AB - Over the past twenty years, Medicare has been transformed from a single-payer insurer into a hybrid of complementary public and private insurance arrangements. Despite creating ongoing controversy, these changes have resulted in an ironic and largely overlooked strategic potential: Medicare's evolving hybrid form makes it the most promising vehicle for overcoming the historical obstacles to universal health insurance in the United States. To make this surprising case, we first explore the distinctive political dynamics of programs that, like today's Medicare, are hybrids of public and private arrangements. We then consider how these political dynamics might circumvent past barriers to universal health insurance. Finally, we discuss the strengths and weaknesses of alternative pathways through which Medicare could be expanded to promote health security. PMID- 17463408 TI - Medicare as incubator for innovation in payment policy. PMID- 17463409 TI - Medicare's politics. PMID- 17463410 TI - Improving the design of competitive bidding in Medicare Advantage. AB - In 2003, Congress passed the Medicare Prescription Drug, Improvement, and Modernization Act, which required that in 2006 the Centers for Medicare and Medicaid Services (CMS) implement a system of competitive bids to set payments for the Medicare Advantage program. Managed care plans now bid for the right to enroll Medicare beneficiaries. Data from the first year of bidding suggest that imperfect competition is limiting the success of the bidding system. This article offers suggestions to improve this system based on findings from auction theory and previous government-run auctions. In particular, CMS can benefit by adjusting its system of competitive bids in four ways: credibly committing to regulations governing bidding; limiting the scope for collusion, entry deterrence, and predatory behavior among bidders; adjusting how benchmark reimbursement rates are set; and accounting for asymmetric information among bidders. PMID- 17463411 TI - The effect of urban air pollution on inflammation, oxidative stress, coagulation, and autonomic dysfunction in young adults. AB - RATIONALE: The biological mechanisms linking air pollution to cardiovascular events still remain largely unclear. OBJECTIVES: To investigate whether biological mechanisms linking air pollution to cardiovascular events occurred concurrently in human subjects exposed to urban air pollutants. METHODS: We recruited a panel of 76 young, healthy students from a university in Taipei. Between April and June of 2004 or 2005, three measurements were made in each participant of high-sensitivity C-reactive protein (hs-CRP), 8-hydroxy-2' deoxyguanosine (8-OHdG), plasminogen activator fibrinogen inhibitor-1 (PAI-1), tissue-type plasminogen activator (tPA) in plasma, and heart rate variability (HRV). Gaseous air pollutants were measured at one air-monitoring station inside their campus, and particulate air pollutants were measured at one particulate matter supersite monitoring station 1 km from their campus. We used linear mixed effects models to associate biological endpoints with individual air pollutants averaged over 1- to 3-day periods before measurements were performed. MEASUREMENTS AND MAIN RESULTS: We found that increases in hs-CRP, 8-OHdG, fibrinogen, and PAI-1, and decreases in HRV indices were associated with increases in levels of particles with aerodynamic diameters less than 10 microm and 2.5 microm, sulfate, nitrate, and ozone (O(3)) in single-pollutant models. The increase in 8-OHdG, fibrinogen, and PAI-1, and the reduction in HRV remained significantly associated with 3-day averaged sulfate and O(3) levels in two pollutant models. There were moderate correlations (r = -0.3) between blood markers of hs-CRP, fibrinogen, PAI-1, and HRV indices. CONCLUSIONS: Urban air pollution is associated with inflammation, oxidative stress, blood coagulation and autonomic dysfunction simultaneously in healthy young humans, with sulfate and O(3) as two major traffic-related pollutants contributing to such effects. PMID- 17463412 TI - Effect of 1-year treatment with roflumilast in severe chronic obstructive pulmonary disease. AB - RATIONALE: The oral phosphodiesterase-4 (PDE4) inhibitor, roflumilast, can improve lung function in moderate chronic obstructive pulmonary disease (COPD). Whether treatment is effective in more severe COPD (GOLD [Global Initiative for Chronic Obstructive Lung Disease] stages III and IV) over a longer period is unknown. OBJECTIVES: To determine whether roflumilast improves lung function and decreases exacerbation frequency over 1 year in patients with stable COPD. METHODS: We conducted a randomized, placebo-controlled, double-blind, parallel group trial for 1 year. We recruited 1,513 patients (mean post-bronchodilator FEV1 41% predicted), 760 receiving oral 500 microg roflumilast and 753 receiving placebo once daily. MEASUREMENTS AND MAIN RESULTS: We recorded post bronchodilator FEV1, exacerbation rate, St. George's Respiratory Questionnaire total score at the study end point, and number and type of reported adverse events during treatment. Post-bronchodilator FEV1 increased by 39 ml with roflumilast compared with placebo by 52 weeks (p=0.001). The mean exacerbation rate was low and comparable in both treatment groups (0.86 vs. 0.92 exacerbations/patient/yr for roflumilast and placebo, respectively). In a retrospective analysis, the exacerbation rate in patients in GOLD stage IV disease was 36% lower in patients treated with roflumilast than in those treated with placebo (1.01 vs. 1.59 exacerbations/patient/year, respectively; p=0.024). The St. George's Respiratory Questionnaire total score did not differ between treatments. The commonest adverse events related to roflumilast treatment were diarrhea, nausea, and headache, which usually subsided during continued treatment. However, roflumilast resulted in more withdrawals within the first 3 to 4 weeks of administration. CONCLUSIONS: In severe, stable COPD, PDE4 inhibition with roflumilast produced a modest but significant improvement in lung function without changing the exacerbation rate or health status. However, patients with very severe disease experienced fewer exacerbations with roflumilast. PMID- 17463413 TI - Airway distensibility in adults with asthma and healthy adults, measured by forced oscillation technique. AB - RATIONALE: Reduced airway distensibility in subjects with asthma compared with control subjects may be related to differences in lung elastic recoil and bronchomotor tone. OBJECTIVES: To examine the contribution of lung elastic recoil and bronchomotor tone to airway distensibility. METHODS: We compared airway distensibility in 18 subjects with asthma with 19 control subjects before and after bronchodilator administration and, in a subgroup of 7 subjects with asthma and 8 control subjects, correlated distensibility with pressure-volume parameters. MEASUREMENTS AND MAIN RESULTS: Distensibility was measured, using the forced oscillation technique, as the linear slope of conductance versus volume between total lung capacity (TLC) and 75% TLC and between 75% TLC and FRC. Transpulmonary pressure was recorded concurrently with distensibility, using an esophageal balloon. Pressure-conductance data were described using linear regressions and pressure-volume data were described using exponential equations. Subjects with asthma had lower baseline FEV1 (p=0.0003) and conductance (p=0.002) than did control subjects. Distensibility above 75% TLC was less in subjects with asthma than in control subjects (p<0.0001), but there was no difference below 75% TLC. Bronchodilator administration did not alter distensibility despite increases in FEV1 (p=0.0002) and conductance (p<0.0001) in subjects with asthma, and conductance (p=0.0004) in control subjects. After bronchodilator administration, subjects with asthma had reduced lung elastic recoil compared with control subjects (p=0.03) and a reduced pressure-conductance slope (p=0.01), but there were no correlations between pressure-volume characteristics and airway distensibility. CONCLUSIONS: Airway distensibility measured by forced oscillation technique is reduced in subjects with asthma compared with subjects without asthma, is not related to lung elastic recoil, and is unchanged by bronchodilator administration. Airway wall remodeling remains the most likely cause of reduced airway distensibility in asthma. PMID- 17463414 TI - Epithelial cell proliferation contributes to airway remodeling in severe asthma. AB - RATIONALE: Despite long-term therapy with corticosteroids, patients with severe asthma develop irreversible airway obstruction. OBJECTIVES: To evaluate if there are structural and functional differences in the airway epithelium in severe asthma associated with airway remodeling. METHODS: In bronchial biopsies from 21 normal subjects, 11 subjects with chronic bronchitis, 9 subjects with mild asthma, and 31 subjects with severe asthma, we evaluated epithelial cell morphology: epithelial thickness, lamina reticularis (LR) thickness, and epithelial desquamation. Levels of retinoblastoma protein (Rb), Ki67, and Bcl-2 were measured, reflecting cellular proliferation and death. Terminal deoxynucleotidyl-mediated dUTP nick end labeling (TUNEL) was used to study cellular apoptosis. MEASUREMENTS AND MAIN RESULTS: Airway epithelial and LR thickness was greater in subjects with severe asthma compared with those with mild asthma, normal subjects, and diseased control subjects (p=0.009 and 0.033, respectively). There was no significant difference in epithelial desquamation between groups. Active, hypophosphorylated Rb expression was decreased (p=0.002) and Ki67 was increased (p<0.01) in the epithelium of subjects with severe asthma as compared with normal subjects, indicating increased cellular proliferation. Bcl-2 expression was decreased (p<0.001), indicating decreased cell death suppression. There was a greater level of apoptotic activity in the airway biopsy in subjects with severe asthma as compared with the normal subjects using the TUNEL assay (p=0.002), suggesting increased cell death. CONCLUSIONS: In subjects with severe asthma, as compared with subjects with mild asthma, normal subjects, and diseased control subjects, we found novel evidence of increased cellular proliferation in the airway contributing to a thickened epithelium and LR. These changes may contribute to the progressive decline in lung function and airway remodeling in patients with severe asthma. PMID- 17463416 TI - IkappaB genetic polymorphisms and invasive pneumococcal disease. AB - RATIONALE: Increasing evidence supports a key role for the transcription factor nuclear factor (NF)-kappaB in the host response to pneumococcal infection. Control of NF-kappaB activity is achieved through interactions with the IkappaB family of inhibitors, encoded by the genes NFKBIA, NFKBIB, and NFKBIE. Rare NFKBIA mutations cause immunodeficiency with severe bacterial infection, raising the possibility that common IkappaB gene polymorphisms confer susceptibility to common bacterial disease. OBJECTIVES: To determine whether polymorphisms in NFKBIA, NFKBIB, and NFKBIE associate with susceptibility to invasive pneumococcal disease (IPD) and thoracic empyema. METHODS: We studied the frequencies of 62 single-nucleotide polymorphisms (SNPs) across NFKBIA, NFKBIB, and NFKBIE in individuals with IPD and control subjects (n=1,060). Significantly associated SNPs were then studied in a group of individuals with thoracic empyema and a second control group (n=632). MEASUREMENTS AND MAIN RESULTS: Two SNPs in the NFKBIA promoter region were associated with protection from IPD in both the initial study group and the pneumococcal empyema subgroup. Significant protection from IPD was observed for carriage of mutant alleles at these two loci on combining the groups (SNP rs3138053: Mantel-Haenszel 2x2 chi2=13.030, p=0.0003; odds ratio [OR], 0.60; 95% confidence interval [CI], 0.45-0.79; rs2233406: Mantel Haenszel 2x2 chi2=18.927, p=0.00001; OR, 0.55; 95% CI, 0.42-0.72). An NFKBIE SNP associated with susceptibility to IPD but not pneumococcal empyema. None of the NFKBIB SNPs associated with IPD susceptibility. CONCLUSIONS: NFKBIA polymorphisms associate with susceptibility to IPD. Genetic variation in an inhibitor of NF kappaB therefore not only causes a very rare immunodeficiency state but may also influence the development of common infectious disease. PMID- 17463415 TI - Arhgef1 is required by T cells for the development of airway hyperreactivity and inflammation. AB - RATIONALE: Arhgef1 is an intracellular protein, expressed by hematopoietic cells, that regulates signaling by both G protein-coupled receptors and RhoA, and, consequently, is required for appropriate migration and adhesion of diverse leukocyte populations. OBJECTIVES: To evaluate a possible contribution for Arhgef1 in the development of airway inflammation and airway hyperreactivity. METHODS: Arhgef1-deficient (Arhgef1-/-) and wild-type (WT) mice were sensitized and airway challenged, followed by measurement of airway responsiveness to inhaled methacholine. Inflammation was assessed by several parameters that included flow cytometric analysis and histology. Arhgef1-deficient recipients were reconstituted with WT T lymphocytes before sensitization and challenge, and again measured for airway responsiveness and inflammation. Cytokine production in response to specific antigen was measured in cultures of isolated leukocytes from lung and spleen and compared with the levels generated in lung and spleen explant cultures. MEASUREMENTS AND MAIN RESULTS: Arhgef1-/- mice display significantly reduced airway hyperreactivity, Th2 cytokine production, and lung inflammation, despite intact systemic immunity. After airway challenge of Arhgef1-/- mice, antigen-specific T cells were present in mutant lungs, but were found to interact with CD11c+ cells at a significantly reduced frequency. Adoptive transfer of WT T cells into Arhgef1-/- mice restored airway hyperreactivity and inflammation. CONCLUSIONS: These data demonstrate that T cells depend on Arhgef1 to promote lung inflammation. Moreover, a deficiency in Arhgef1 results in reduced T cell CD11c+ antigen-presenting cell interaction, and likely underscores the inability of Arhgef1-/- mice to mount an adaptive immune response to airway challenge. PMID- 17463417 TI - Proangiogenic activity in bronchoalveolar lavage fluid from patients with asthma. AB - RATIONALE: Asthmatic airways have an increased number and size of vascular structures, which contribute to airflow obstruction and hyperresponsiveness. OBJECTIVES: We examined whether proangiogenic mediators are elevated in bronchoalveolar lavage fluid (BALF) from subjects with asthma and if this translated to induction of angiogenesis. METHODS: Angiogenic activity in BALF from 12 healthy, nonatopic subjects and 10 atopic subjects with mild asthma was evaluated by examining tubule formation at 11 days in cocultures of human endothelial cells with dermal fibroblasts. Vascular structures were visualized by anti-CD31 labeling and quantified by image analysis. Angiogenic growth factors in BALF from healthy subjects and subjects with asthma were identified using antibody arrays and by ELISA. MEASUREMENTS AND MAIN RESULTS: Angiogenic activity induced by BALF from healthy subjects was not different from basal tubule formation (p>0.05). However, induction of tubular structures by asthmatic BALF was 2.5-fold greater (p<0.001) compared with healthy samples. Similarly, levels of proangiogenic growth factors (angiogenin, vascular endothelial growth factor [VEGF], monocyte chemotactic protein-1) were increased approximately 2.5-fold (p<0.05) in BALF from subjects with asthma, whereas antiangiogenic factors (endostatin, Ang-2) were unchanged. A blocking anti-VEGF antibody abolished tubule formation induced by BALF from either healthy subjects or subjects with asthma (p<0.01). Immunodepletion of VEGF had no effect on basal tubule formation induced by healthy BALF but abrogated enhanced tubule formation by asthmatic BALF (p<0.01). CONCLUSIONS: BALF collected from subjects with asthma but not healthy subjects is functionally active in promoting angiogenesis in vitro. The proangiogenic capacity of BALF from subjects with asthma resides in elevated VEGF derived from asthmatic airways. This observation supports VEGF as a key factor in vascular remodeling in asthma. PMID- 17463418 TI - A single dose of vitamin D enhances immunity to mycobacteria. AB - RATIONALE: Vitamin D was used to treat tuberculosis (TB) in the preantibiotic era. Prospective studies to evaluate the effect of vitamin D supplementation on antimycobacterial immunity have not previously been performed. OBJECTIVES: To determine the effect of vitamin D supplementation on antimycobacterial immunity and vitamin D status. METHODS: A double-blind randomized controlled trial was conducted in 192 healthy adult TB contacts in London, United Kingdom. Participants were randomized to receive a single oral dose of 2.5 mg vitamin D or placebo and followed up at 6 weeks. MEASUREMENTS AND MAIN RESULTS: The primary outcome measure was assessed with a functional whole blood assay (BCG-lux assay), which measures the ability of whole blood to restrict luminescence, and thus growth, of recombinant reporter mycobacteria in vitro; the readout is expressed as a luminescence ratio (luminescence postinfection/baseline luminescence). IFN gamma responses to the Mycobacterium tuberculosis antigens early secretory antigenic target-6 and culture filtrate protein 10 were determined with a second whole blood assay. Vitamin D supplementation significantly enhanced the ability of participants' whole blood to restrict BCG-lux luminescence in vitro compared with placebo (mean luminescence ratio at follow-up, 0.57, vs. 0.71, respectively; 95% confidence interval for difference, 0.01-0.25; p=0.03) but did not affect antigen-stimulated IFN-gamma secretion. CONCLUSIONS: A single oral dose of 2.5 mg vitamin D significantly enhanced the ability of participants' whole blood to restrict BCG-lux luminescence in vitro without affecting antigen-stimulated IFN gamma responses. Clinical trials should be performed to determine whether vitamin D supplementation prevents reactivation of latent TB infection. Clinical trial registered with www.clinicaltrials.gov (NCT 00157066). PMID- 17463419 TI - Standard reference for the six-minute-walk test in healthy children aged 7 to 16 years. AB - RATIONALE: We have previously reported that the six-minute-walk test (6MWT) is a reliable and valid functional test for assessing exercise tolerance and endurance. There is a lack of pediatric standard reference for the 6MWT. OBJECTIVES: To construct height-specific standards for the 6MWT for children aged 7 to 16 years. METHODS: The anthropometric data, spirometric lung function, and six-minute-walk distance (6MWD) of Chinese children aged 7 to 16 years were prospectively measured using standardized protocols. The findings were used to construct height-specific standards for the 6MWT. The least mean square (LMS) method using maximum penalized likelihood was used to facilitate model fitting. Factors significantly associated with 6MWD were also determined. MEASUREMENTS AND MAIN RESULTS: From January 2005 to April 2006, a total of 1,445 subjects were studied. The measured variables showed a normal distribution. Height-specific reference standards for 6MWT were constructed for both male and female children. Forward stepwise multiple regression analysis revealed height and difference in heart rate before and after the walk test to be important clinical variables associated with 6MWD. CONCLUSIONS: These 6MWD standards will provide useful references for the care of pediatric patients. PMID- 17463420 TI - Salt and cardiovascular disease. PMID- 17463421 TI - Postmenopausal hormone therapy. PMID- 17463422 TI - Preventing ventilator associated pneumonia. PMID- 17463423 TI - Public health policy must be based on sound evidence. PMID- 17463424 TI - For debate: Should prostitution be legalised and regulated? PMID- 17463425 TI - BMA public health doctor's claims lack evidence. PMID- 17463426 TI - Shift work: The view from Denmark. PMID- 17463427 TI - Coeliac disease: Decision tool needs to be developed for children. PMID- 17463428 TI - Drug management of COPD. PMID- 17463430 TI - Exodus of medical staff strains Iraq's health facilities. PMID- 17463431 TI - New contract has meant higher pay but not better care, say doctors. PMID- 17463433 TI - US Supreme Court approves ban on "partial birth abortion". PMID- 17463435 TI - Abstinence education has no effect on US teenagers' sexual activity. PMID- 17463434 TI - Report recommends tighter legislation and better road design to reduce traffic injuries and deaths. PMID- 17463436 TI - Hospital patients should be assessed for risk of thromboembolism. PMID- 17463437 TI - Inquiry will study claims that Sellafield workers' body parts were removed without families' consent. PMID- 17463440 TI - UK leads initiative to drive down cost of drugs in poor countries. PMID- 17463442 TI - Group of British doctors call for a boycott of the Israel Medical Association. PMID- 17463444 TI - UK stillbirth rate stops declining as age of mothers rises. PMID- 17463445 TI - NICE tells employers how to help staff quit smoking. PMID- 17463446 TI - First steps are taken towards creation of a European digital library. PMID- 17463447 TI - Working within industry's silken but firm embrace. PMID- 17463456 TI - Global health: Great expectations. PMID- 17463457 TI - NHS finances: Farewell to dodging and weaving. PMID- 17463458 TI - Bad blood. PMID- 17463459 TI - Why single-payer health systems spark endless debate. PMID- 17463460 TI - Waking up from the DREAM of preventing diabetes with drugs. PMID- 17463461 TI - Anorexia nervosa. PMID- 17463462 TI - Acute dystonic reaction caused by metoclopramide, versus tetanus. PMID- 17463463 TI - Dipyridamole with aspirin is better than aspirin alone in preventing vascular events after ischaemic stroke or TIA. PMID- 17463464 TI - Statins, saving lives, and shibboleths. PMID- 17463465 TI - Myths and realities. PMID- 17463466 TI - Medicine: more than a career for my daughter. PMID- 17463467 TI - Wit. PMID- 17463469 TI - In vitro and in silico analysis of annexin V binding to lymphocytes as a biomarker in emergency department sepsis studies. AB - BACKGROUND: Peripheral blood lymphocyte apoptosis is a recognized feature of serious infection and sepsis and can be easily quantified by flow cytometric measurement of annexin V binding to the cell surface. Use of apoptosis as a biomarker in emergency department (ED) studies of sepsis is potentially difficult because of sample processing requirements and limited availability of a research cytometer with which to measure patient samples. OBJECTIVES: To assess, in vitro and in simulation, the relationship between sample stability, timing of patient enrollment, and diagnostic performance of a flow cytometric assay for sepsis in patients evaluated in EDs. METHODS: Assuming any clinical trial would require daily sample batching, the authors measured the stability of lymphocyte samples over time, noting the rate at which annexin V-negative cells became positive as ED processing delays increased. With these data, they then optimized a study design that could evaluate lymphocyte apoptosis as a sepsis biomarker by using a series of Monte Carlo-based simulated clinical trials. RESULTS: The authors found that annexin V-negative lymphocytes become positive during storage delays that would be encountered in an ED sepsis trial. The extent of this deterioration was least among cells left as whole blood at room temperature until just before analysis or when lymphocytes were isolated early and stored in culture media at 4 degrees C until analysis. When the expected rate of sample deterioration was considered in simulated clinical trials, an inverse relationship was found between the rate at which patients are enrolled and the best achievable receiver operating characteristic curve a study could produce. CONCLUSIONS: Peripheral blood samples being analyzed for lymphocyte apoptosis degrade at a rate relevant to the design of ED trials of sepsis. Because of sample processing delays inherent in studying unscheduled septic patients, the performance of annexin V binding as a biomarker for sepsis can approach, but not be expected to exceed, its performance in a comparable intensive care unit-based study. PMID- 17463470 TI - Abstracts of the 2007 Society of Academic Emergency Medicine (SAEM) Annual Meeting. PMID- 17464044 TI - Nonribosomal peptide synthesis in Aspergillus fumigatus and other fungi. AB - In fungi, nonribosomal peptide synthetases (NRP synthetases) are large multi functional enzymes containing adenylation, thiolation (or peptidyl carrier protein, PCP) and condensation domains. These enzymes are often encoded within gene clusters. Multiple NRP synthetase ORFs have also been identified in fungi (14 in Aspergillus fumigatus). LeaA, a methyltransferase, is involved in secondary metabolite gene cluster regulation in Aspergillus spp. The NRP synthetases GliP and FtmA respectively direct the biosynthesis of the toxic metabolites gliotoxin and brevianamide F, a precursor of bioactive prenylated alkaloids. The NRP synthetase Pes1 has been shown to mediate resistance to oxidative stress, and in plant-pathogenic ascomycetes (e.g. Cochliobolus heterostrophus) an NRP synthetase, encoded by the NPS6 gene, significantly contributes to virulence and resistance to oxidative stress. Adenylation (A) domains within NRP synthetases govern the specificity of amino acid incorporation into nonribosomally synthesized peptides. To date there have only been limited demonstrations of A domain specificity (e.g. A. fumigatus GliP and in Beauveria bassiana) in fungi. Indeed, only in silico prediction data are available on A domain specificity of NRP synthetases from most fungi. NRP synthetases are activated by 4'-phosphopantetheinylation of serine residues within PCP domains by 4'-phosphopantetheinyl transferases (4'-PPTases). Coenzyme A acts as the 4' phosphopantetheine donor, and labelled coenzyme A can be used to affinity-label apo-NRP synthetases. Emerging fungal gene disruption and gene cluster expression strategies, allied to proteomic strategies, are poised to facilitate a greater understanding of the coding potential of NRP synthetases in fungi. PMID- 17464045 TI - Differential gene expression profiling of Streptococcus mutans cultured under biofilm and planktonic conditions. AB - Streptococcus mutans often adopts a sessile biofilm lifestyle that differs greatly from that of free-living cells. Biofilm formation represents a protected mode of growth that allows cells to survive in hostile environments. In this study, in vitro comparative transcriptome analysis was carried out to identify genes that are differentially expressed in biofilm of S. mutans compared with free-living cells. DNA-microarray analyses indicated that about 12 % of genes showed significant differential expression: 139 were activated and 104 were repressed in biofilm vs the planktonic environment. The differential expression of 20 selected genes was confirmed by real-time RT-PCR. In addition, regulation of expression of these genes during biofilm development was tested in 100 and 400 microm deep biofilms. Direct comparison of optical images consistently demonstrated that changes in biofilm thickness are accompanied by significant shifts in cell viability. From evaluation of gene expression patterns, it was shown that the majority of the genes tested were significantly down-regulated in 400 vs 100 microm deep biofilms. This study provides a genome-scale synopsis and adds important insights into gene expression in biofilm development processes of S. mutans, which are strongly associated with the pathogenesis of dental diseases. PMID- 17464046 TI - Effects of iron on DNA release and biofilm development by Pseudomonas aeruginosa. AB - Extracellular DNA is one of the major matrix components in Pseudomonas aeruginosa biofilms. It functions as an intercellular connector and plays a role in stabilization of the biofilms. Evidence that DNA release in P. aeruginosa PAO1 biofilms is controlled by the las-rhl and pqs quorum-sensing systems has been previously presented. This paper provides evidence that DNA release in P. aeruginosa PAO1 biofilms is also under iron regulation. Experiments involving cultivation of P. aeruginosa in microtitre trays suggested that pqs expression, DNA release and biofilm formation were favoured in media with low iron concentrations (5 microM FeCl(3)), and decreased with increasing iron concentrations. Experiments involving cultivation of P. aeruginosa in a flow chamber system suggested that a high level of iron (100 microM FeCl(3)) in the medium suppressed DNA release, structural biofilm development, and the development of subpopulations with increased tolerance toward antimicrobial compounds. Experiments with P. aeruginosa strains harbouring fluorescent reporters suggested that expression of the pqs operon was induced in particular subpopulations of the biofilm cells under low-iron conditions (1 microM FeCl(3)), but repressed in the biofilm cells under high-iron conditions (100 microM FeCl(3)). PMID- 17464047 TI - Rapid necrotic killing of polymorphonuclear leukocytes is caused by quorum sensing-controlled production of rhamnolipid by Pseudomonas aeruginosa. AB - Quorum sensing (QS) denotes a density-dependent mode of inter-bacterial communication based on signal transmitter molecules. Active QS is present during chronic infections with the opportunistic pathogen Pseudomonas aeruginosa in immunocompromised patients. The authors have previously demonstrated a QS regulated tolerance of biofilm bacteria to the antimicrobial properties of polymorphonuclear leukocytes (PMNs). The precise QS-regulated effect on the PMNs is, however, unknown. Incubation of human PMNs with supernatants from dense P. aeruginosa cultures showed that the QS-competent P. aeruginosa induced rapid necrosis of the PMNs. This mechanism was also observed in mouse lungs infected with P. aeruginosa, and in sputum obtained from P.-aeruginosa-infected patients with cystic fibrosis. Evidence is presented that the necrotic effect was caused by rhamnolipids, production of which is QS controlled. The results demonstrate the potential of the QS system to facilitate infections with P. aeruginosa by disabling the PMNs, which are a major first line of defence of the host. Furthermore, the study emphasizes the inhibition of QS as a target for the treatment of infections with P. aeruginosa. PMID- 17464048 TI - An aflagellate mutant Yersinia enterocolitica biotype 1A strain displays altered invasion of epithelial cells, persistence in macrophages, and cytokine secretion profiles in vitro. AB - Despite being classically defined as non-pathogenic, there is growing evidence that biotype 1A Yersinia enterocolitica isolates may be aetiological agents of disease in humans. In previous studies, a potential link between motility and the ability of biotype 1A strains to invade cultured epithelial cells was observed. In an attempt to further investigate this finding, a flagella mutant was constructed in a human faecal Y. enterocolitica biotype 1A isolate. The flagella mutation abolished the ability of the strain to invade cultured human epithelial cells, although adherence was not affected. The aflagellate mutant was also attenuated in its ability to survive within cultured macrophages, being cleared after 3 h, whilst the wild-type persisted for 24 h after infection. Examination of cytokine secretion by infected macrophages also suggested that the flagella of biotype 1A strains act as anti-inflammatory agents, decreasing production of tumour necrosis factor (TNF)-alpha whilst increasing secretion of interleukin (IL)-10. Preliminary studies using porcine in vitro organ culture (IVOC) tissue suggested that the flagella mutant was also attenuated in its ability to colonize intestinal tissue. PMID- 17464049 TI - Analysis of the expression, regulation and export of NleA-E in Escherichia coli O157 : H7. AB - Previous work has shown that locus of enterocyte effacement (LEE)-encoded effector proteins such as Tir and Map can be exported via the type III secretion system (T3SS) of Escherichia coli O157 : H7. Additionally, a family of non-LEE encoded (Nle) effector proteins has been shown to be secreted from Citrobacter rodentium, homologues of which are located on the E. coli O157 chromosome. While NleA has been shown to be secreted from pathogenic E. coli, the secretion of other Nle effector proteins has only been detected under induced conditions, or using a mutated T3SS. This study aimed to determine: (1) which nle genes are expressed in E. coli O157 : H7 under secretion-permissive conditions; (2) if Nle proteins are secreted from wild-type E. coli O157 : H7 under secretion-permissive conditions; and (3) if nle gene expression is regulated co-ordinately with other LEE-encoded effectors. Using data generated from a combination of transcriptome arrays, reporter fusions and proteomics, it was demonstrated that only nleA is expressed co-ordinately with the LEE. Secretion and expression of NleA were regulated directly or indirectly by ler, a key activator of the LEE. MS confirmed the secretion of NleA into the culture supernatant, while NleB-F were not detected. PMID- 17464050 TI - Regulated synthesis of the Borrelia burgdorferi inner-membrane lipoprotein IpLA7 (P22, P22-A) during the Lyme disease spirochaete's mammal-tick infectious cycle. AB - Results of previous immunological studies suggested that Borrelia burgdorferi regulates synthesis of the IpLA7 lipoprotein during mammalian infection. Through combined use of quantitative reverse transcription PCR, immunofluorescence analyses, ELISA and immunoblotting, it is now demonstrated that IpLA7 is actually expressed throughout mammalian infection, as well as during transmission both from feeding ticks to naive mice and from infected mice to naive, feeding ticks. However, proportions of IpLA7-expressing B. burgdorferi within tick midguts declined significantly with time following completion of blood feeding. Cultured bacteria differentially expressed IpLA7 in response to changes in temperature, pH and concentration of 4,5-dihydroxy-2,3-pentanedione, the precursor of autoinducer 2, indicative of mechanisms governing IpLA7 expression. Previous studies also reported mixed results as to the cellular localization of IpLA7. It is now demonstrated that IpLA7 localizes primarily to the borrelial inner membrane and is not surface-exposed, consistent with the ability of these bacteria to produce IpLA7 throughout mammalian infection despite being the target of a robust immune response. PMID- 17464051 TI - Disruption of the Candida albicans ATC1 gene encoding a cell-linked acid trehalase decreases hypha formation and infectivity without affecting resistance to oxidative stress. AB - In Candida albicans, the ATC1 gene, encoding a cell wall-associated acid trehalase, has been considered as a potentially interesting target in the search for new antifungal compounds. A phenotypic characterization of the double disruptant atc1Delta/atc1Delta mutant showed that it was unable to grow on exogenous trehalose as sole carbon source. Unlike actively growing cells from the parental strain (CAI4), the atc1Delta null mutant displayed higher resistance to environmental insults, such as heat shock (42 degrees C) or saline exposure (0.5 M NaCl), and to both mild and severe oxidative stress (5 and 50 mM H(2)O(2)), which are relevant during in vivo infections. Parallel measurements of intracellular trehalose and trehalose-metabolizing enzymes revealed that significant amounts of the disaccharide were stored in response to thermal and oxidative challenge in the two cell types. The antioxidant activities of catalase and glutathione reductase were triggered by moderate oxidative exposure (5 mM H(2)O(2)), whereas superoxide dismutase was inhibited dramatically by H(2)O(2), where a more marked decrease was observed in atc1Delta cells. In turn, the atc1Delta mutant exhibited a decreased capacity of hypha and pseudohypha formation tested in different media. Finally, the homozygous null mutant in a mouse model of systemic candidiasis displayed strongly reduced pathogenicity compared with parental or heterozygous strains. These results suggest not only a novel role for the ATC1 gene in dimorphism and infectivity, but also that an interconnection between stress resistance, dimorphic conversion and virulence in C. albicans may be reconsidered. They also support the hypothesis that Atc1p is not involved in the physiological hydrolysis of endogenous trehalose. PMID- 17464052 TI - Structural analysis of a non-ribosomal halogenated cyclic peptide and its putative operon from Microcystis: implications for evolution of cyanopeptolins. AB - The structure of the major peptide produced by Microcystis cf. wesenbergii NIVA CYA 172/5, the halogenated heptapeptide cyanopeptolin-984, was determined using LC/MS/MS. A gene cluster encoding a peptide synthetase putatively producing a cyanopeptolin was cloned from the same strain and sequenced. The cluster consists of four genes encoding peptide synthetases and one gene encoding a halogenase. Two additional ORFs transcribed in the opposite direction were found in the 5' flanking sequence; one of these encodes an ABC transporter. The overall organization of the cyanopeptolin synthetase operon (mcn) resembles a previously analysed anabaenopeptilide synthetase operon (apd) from Anabaena strain 90. Phylogenetic analyses of the individual domains from Mcn, Apd and other cyanobacterial peptide synthetases showed clustering of the adenylation domains according to function irrespective of operon origin - indicating strong functional constraints across peptide synthetases. In contrast, the condensation and thiolation domains to a large extent grouped according to operon affiliation or position in the respective operons. Phylogenetic analyses of condensation domains indicated that N-terminal domains and domains that condense L-amino acids and D-amino acids, respectively, form three separate groups. Although recombination events are likely to be involved in the evolution of mcn, no clear evidence of genetic recombination between the two cyanopeptolin gene clusters was found. Within the genus Microcystis, microcystin and cyanopeptolin synthetases have an evolutionary history of genomic coexistence. However, the data indicated that the two classes of peptide synthetase gene clusters have evolved independently. PMID- 17464053 TI - ScbA from Streptomyces coelicolor A3(2) has homology to fatty acid synthases and is able to synthesize gamma-butyrolactones. AB - gamma-Butyrolactones play an important role in the regulation of antibiotic production and differentiation in Streptomyces. However the biosynthetic pathway for these small molecules has not yet been determined, and in vitro synthesis has not been reported. The function of the AfsA family of proteins, originally proposed to catalyse gamma-butyrolactone synthesis, has been in debate. To clarify the function of the AfsA family, and to understand the synthesis of the gamma-butyrolactones, we performed in silico analysis of this protein family. AfsA proteins consist of two divergent domains, each of which has similarity to the fatty acid synthesis enzymes FabA and FabZ. The two predicted active sites in ScbA, which is the AfsA orthologue found in Streptomyces coelicolor, were mutated, and gamma-butyrolactone biosynthesis was abolished in all four constructed mutants, strongly suggesting that ScbA has enzymic activity. PMID- 17464054 TI - MbtH-like protein-mediated cross-talk between non-ribosomal peptide antibiotic and siderophore biosynthetic pathways in Streptomyces coelicolor M145. AB - MbtH-like proteins are a family of small proteins encoded by genes found in many, but not all, non-ribosomal peptide synthetase-encoding gene clusters that direct the biosynthesis of peptide antibiotics and siderophores. Studies published to date have not elucidated the function of MbtH-like proteins, nor have they clarified whether they are required for metabolite biosynthesis. Here it is shown that only one of two genes (cdaX or cchK) encoding MbtH-like proteins in Streptomyces coelicolor is required for biosynthesis of the peptide siderophore coelichelin and the calcium-dependent peptide antibiotic (CDA). The cdaX and cchK genes can functionally complement each other in trans, suggesting that CdaX and CchK can cross-talk with the coelichelin and CDA biosynthetic pathways, respectively. Transcriptional analyses of wild-type S. coelicolor and a double cchK/cdaX replacement mutant indicate that CchK and CdaX may not be involved in transcriptional regulation of coelichelin and CDA biosynthetic gene clusters. PMID- 17464055 TI - Effects of deletions of mbtH-like genes on clorobiocin biosynthesis in Streptomyces coelicolor. AB - In the biosynthetic gene cluster of the aminocoumarin antibiotic clorobiocin, the small ORF cloY encodes a 71 aa protein which shows significant sequence similarity to mbtH from the mycobactin biosynthetic gene cluster of Mycobacterium tuberculosis. mbtH-like genes are frequently found in the biosynthetic gene clusters of peptide antibiotics and siderophores, but their function has remained enigmatic. In a recent publication it has been suggested that these genes may have no function for secondary metabolite biosynthesis. An in-frame deletion of cloY in the clorobiocin cluster has now been carried out. When the modified cluster was expressed in the heterologous host Streptomyces coelicolor M512, clorobiocin was still formed. However, when the two further mbtH-like genes from elsewhere in the host genome were inactivated as well, clorobiocin formation was reduced dramatically. Complementation with cloY or with any of three other mbtH like genes restored clorobiocin formation. This is the first report proving the requirement of an mbtH-like gene for secondary metabolite formation, and the first proof that different mbtH-like genes can functionally replace each other. Feeding of an mbtH-defective triple mutant strain with an intact 3-amino-4,7 dihydroxy-coumarin moiety restored antibiotic production, showing that cloY is specifically required for the formation of this moiety of the clorobiocin molecule. PMID- 17464056 TI - The key role of the mycolic acid content in the functionality of the cell wall permeability barrier in Corynebacterineae. AB - Recently, it has been shown that trehalose and mycolic acids are essential for the growth of Mycobacterium tuberculosis, the causative agent of tuberculosis, and Mycobacterium smegmatis, and important but not indispensable to the survival of Corynebacterium glutamicum. Therefore, to investigate the function of mycolic acids in both the permeability of the cell wall to small nutrients and antibiotics, and the excretion of amino acids by C. glutamicum, a trehalose deficient mutant of the L-lysine producer ATCC 21527, designated LP Delta treS Delta otsA Delta treY, was constructed. By using different carbon sources in either the presence or the absence of external trehalose, a set of endogenously trehalose-free LP Delta treS Delta otsA Delta treY cells that exhibited various mycolate contents was generated. The results showed that the structure of the arabinogalactan of these different cell types of LP Delta treS Delta otsA Delta treY was not affected when the mycolic acid layer was either missing or impaired. Nevertheless, cells were more susceptible to antibiotics, and the permeability of their cell walls to glycerol was increased. Interestingly, a concomitant increase in the excretion of both L-lysine and L-glutamate was also observed, indicating that the mycolic acid content of the permeability barrier (and not only the peptidoglycan and/or the arabinogalactan) is implicated in the glutamate excretion process. PMID- 17464057 TI - Lipid composition and transcriptional response of Mycobacterium tuberculosis grown under iron-limitation in continuous culture: identification of a novel wax ester. AB - The low level of available iron in vivo is a major obstacle for microbial pathogens and is a stimulus for the expression of virulence genes. In this study, Mycobacterium tuberculosis H37Rv was grown aerobically in the presence of limited iron availability in chemostat culture to determine the physiological response of the organism to iron-limitation. A previously unidentified wax ester accumulated under iron-limited growth, and changes in the abundance of triacylglycerol and menaquinone were also observed between iron-replete and iron-limited chemostat cultures. DNA microarray analysis revealed differential expression of genes involved in glycerolipid metabolism and isoprenoid quinone biosynthesis, providing some insight into the underlying genetic changes that correlate with cell-wall lipid profiles of M. tuberculosis growing in an iron-limited environment. PMID- 17464058 TI - Biological cost of hypermutation in Pseudomonas aeruginosa strains from patients with cystic fibrosis. AB - The high prevalence of hypermutable (mismatch repair-deficient) Pseudomonas aeruginosa strains in patients with cystic fibrosis (CF) is thought to be driven by their co-selection with adaptive mutations required for long-term persistence. Whether the increased mutation rate of naturally hypermutable strains is associated with a biological benefit or cost for the colonization of secondary environments is not known. Thirty-nine P. aeruginosa strains were collected from ten patients with CF during their course of chronic lung infections and screened for hypermutability. Seven hypermutable P. aeruginosa strains (18 %) isolated from six patients with CF (60 %) were identified and assigned to five different genotypes. Complementation and sequence analysis in the mutS, mutL and uvrD genes of these hypermutable P. aeruginosa strains revealed novel mutations. To understand the consequences of hypermutation for the fitness of the organisms, five pairs of clinical wild-type/hypermutable, clonally related P. aeruginosa strains and the laboratory strains PAO1/PAO1DeltamutS were subjected to competition in vitro and in the agar-beads mouse model of chronic airway infection. When tested in competition assay in vitro, the wild-type outcompeted four clinical hypermutable strains and the PAO1DeltamutS strain. In vivo, all of the hypermutable strains were less efficient at establishing lung infection than their wild-type clones. These results suggest that P. aeruginosa hypermutation is associated with a biological cost, reducing the potential for colonization of new environments and therefore strain transmissibility. PMID- 17464059 TI - The Mycoplasma gallisepticum OsmC-like protein MG1142 resides on the cell surface and binds heparin. AB - Mycoplasma gallisepticum is an avian pathogen that causes a chronic respiratory disease of chickens and results in significant economic losses to the poultry industry worldwide. Colonization of the host and the establishment of chronic disease are initiated by the cytadherence of M. gallisepticum to the host respiratory epithelium. While several proteins involved in cytadhesion have been characterized, molecules that interact with components of the host extracellular matrix, a process that is central to pathogenesis, are only now being identified. In this study, M. gallisepticum whole cells were shown to bind heparin in a specific and saturable manner. Heparin also significantly inhibited the binding of M. gallisepticum to the human lung fibroblast cell line MRC-5, suggesting a potential role for glycosaminoglycans (GAGs) in cytadherence. M. gallisepticum protein MG1142 (encoded by mga 1142), which displays homology to the osmotically induced (OsmC) family of proteins, binds strongly to heparin, is highly expressed during in vitro culture, and is surface accessible. Recombinant MG1142 bound heparin in a dose-dependent and saturable manner with a dissociation constant (K(d)) of 10+/-1.8 nM, which is within a physiologically significant range, compared to that of other heparin-binding proteins. Binding to heparin was inhibited by the heavily sulfated polysaccharide fucoidan, but not by mucin or chondroitin sulfate A or B, suggesting that electrostatic interactions between the sulfate groups of heparin and the positively charged basic residues of the MG1142 protein are important in binding. The ability of M. gallisepticum to bind GAGs may contribute to host adherence and colonization. PMID- 17464060 TI - Phenotypic characterization of a virulence-associated protein, VagH, of Yersinia pseudotuberculosis reveals a tight link between VagH and the type III secretion system. AB - Recently, a number of attenuated mutants of Yersinia pseudotuberculosis have been identified using a bioinformatics approach. One of the target genes identified in that study was vagH, which the authors now characterized further. VagH shows homology to HemK of Escherichia coli, possessing methyltransferase activity similar to that of HemK, and targeting release factors 1 and 2. Microarray studies comparing the wild-type and the vagH mutant revealed that the mRNA levels of only a few genes were altered in the mutant. By proteome analysis, expression of the virulence determinant YopD was found to be increased, indicating a possible connection between VagH and the virulence plasmid-encoded type III secretion system (T3SS). Further analysis showed that Yop expression and secretion were repressed in a vagH mutant. This phenotype could be suppressed by trans-complementation with the wild-type vagH gene or by deletion of the negative regulator yopD. Also, in a similar manner to a T3SS-negative mutant, the avirulent vagH mutant was rapidly cleared from Peyer's patches and could not reach the spleen after oral infection of mice. In a manner analogous to that of T3SS mutants, the vagH mutant could not block phagocytosis by macrophages. However, a vagH mutant showed no defects in the T3SS-independent ability to proliferate intracellularly and replicated to levels similar to those of the wild type in macrophages. In conclusion, the vagH mutant exhibits a virulence phenotype similar to that of a T3SS-negative mutant, indicating a tight link between VagH and type III secretion in Y. pseudotuberculosis. PMID- 17464061 TI - Interaction between the Helicobacter pylori accessory proteins HypA and UreE is needed for urease maturation. AB - Several accessory proteins are required for the maturation of two nickel containing enzymes in the gastric pathogen Helicobacter pylori. These two enzymes are hydrogenase and urease. Among the accessory/maturation proteins, the nickel binding HypA protein has been previously shown to be required for the full activity of both the hydrogenase and the urease enzymes, while another nickel binding protein, UreE, is known to be solely involved in the urease maturation process. In this study, UreE was shown to be required under all nickel levels for full activation of the apourease. By use of cross-linking studies, an interaction between purified HypA and UreE proteins was identified, leading to the formation of a 34 kDa heterodimer complex. The cross-linked adduct was detected by immunoblotting with either anti-HypA or anti-UreE antiserum. By using a two plasmid system in Escherichia coli, the highest urease activity was achieved under low nickel conditions only when the UreE protein was expressed along with the wild-type HypA protein, but not with its nickel-binding-deficient variant HypA H2A. Addition of only 1 microM NiCl(2) into minimal medium abolished the need for HypA to activate the urease. Although various attempts to show direct nickel transfer from HypA to UreE failed, these results suggest that interactions between the nickel-binding accessory proteins HypA and UreE are required to allow nickel transfer from HypA eventually to the apourease in H. pylori. PMID- 17464062 TI - Analysis of the Mycobacterium ulcerans genome sequence reveals new loci for variable number tandem repeats (VNTR) typing. AB - Screening of the genome sequence of the Mycobacterium ulcerans strain Agy99 from Ghana with tandem repeats finder software revealed 34 novel non-degenerate tandem repeats containing loci suitable for variable number tandem repeats (VNTR) typing. All loci revealed polymorphism within M. ulcerans isolates of geographically diverse origins. The results confirm the evolutionary scenario suggested by multi-locus sequence typing in which a progenitor of all M. ulcerans lineages emerged from the environmental species Mycobacterium marinum and subsequently diverged into several geographical lineages. For further attempts to develop a VNTR-based genetic fingerprinting tool for M. ulcerans, it is suggested that the focus should rather be on M. marinum than on the African M. ulcerans Agy99 genome sequence as a starting point. PMID- 17464064 TI - The Escherichia coli yhjA gene, encoding a predicted cytochrome c peroxidase, is regulated by FNR and OxyR. AB - The Escherichia coli FNR protein is an oxygen-responsive global transcription factor, and OxyR is a key regulator of the peroxide stress response. Here both FNR and OxyR are shown to regulate expression of the E. coli yhjA gene. The yhjA gene encodes a predicted cytochrome c peroxidase, a bacterial haem-containing protein involved in the peroxide stress response through its ability to convert hydrogen peroxide to water. It is shown that the yhjA gene of E. coli possesses a class II FNR site and an OxyR site upstream of the yhjA transcript start. Expression of yhjA was found to be dependent on this unusual combination of FNR and OxyR under conditions of oxygen starvation. Phenotypic analysis of the yhjA mutant revealed increased sensitivity to exogenous hydrogen peroxide and organic peroxides during growth under anaerobic conditions, consistent with the observed regulation and predicted function of the yhjA gene product. PMID- 17464065 TI - Ferripyochelin uptake genes are involved in pyochelin-mediated signalling in Pseudomonas aeruginosa. AB - In response to iron starvation, Pseudomonas aeruginosa produces the siderophore pyochelin. When secreted to the extracellular environment, pyochelin chelates iron and transports it to the bacterial cytoplasm via its specific outer-membrane receptor FptA and the inner-membrane permease FptX. Exogenously added pyochelin also acts as a signal which induces the expression of the pyochelin biosynthesis and uptake genes by activating PchR, a cytoplasmic regulatory protein of the AraC/XylS family. The importance of ferripyochelin uptake genes in this regulation was evaluated. The fptA and fptX genes were shown to be part of the fptABCX ferripyochelin transport operon, which is conserved in Burkholderia sp. and Rhodospirillum rubrum. The fptB and fptC genes were found to be dispensable for utilization of pyochelin as an iron source, for signalling and for pyochelin production. By contrast, mutations in fptA and fptX not only interfered with pyochelin utilization, but also affected signalling and diminished siderophore production. It is concluded from this that pyochelin-mediated signalling operates to a large extent via the ferripyochelin transport system. PMID- 17464066 TI - Transduction of centrifugation-induced gravity forces through mitogen-activated protein kinase pathways in the fission yeast Schizosaccharomyces pombe. AB - Centrifugation of cells of Schizosaccharomyces pombe in liquid medium prompted a marked activation of Sty1 and Pmk1, which are the effector mitogen-activated protein kinases (MAPKs) of the stress-activated protein kinase pathway and the cell-integrity pathway, respectively. Transduction of the centrifugation signals showed a sensitivity threshold above which the response was dependent on time and temperature. Centrifugation-induced phosphorylation of Sty1 and Pmk1 required the presence of the main functional components of the respective signalling cascades, i.e. Wak1 or Win1 plus Wis1, and Mkh1 plus Pek1. The transcription factor Atf1 also became phosphorylated in a Sty1-dependent way upon centrifugation. Hypergravity was an important factor in the activation of Sty1 induced by centrifugation, whilst activation of Pmk1 was mostly due to gravity-associated shear forces. Centrifugation did not increase cell survival against other stresses. Rather, the increased gravitational forces produced a delay in the cell cycle, probably related to alterations in the actin-polarization pattern. Phosphorylation of the MAPK Sty1 was needed for the depolarization of actin patches induced by the centrifugation stress. PMID- 17464067 TI - Turgor regulation in the osmosensitive cut mutant of Neurospora crassa. AB - The internal hydrostatic pressure (turgor) of fungal cells is maintained at 400 500 kPa. The turgor is regulated by changes in ion flux and by production of the osmotically active metabolite glycerol. In Neurospora crassa, there are at least two genetically distinct pathways that function in adaptation to hyperosmotic shock. One involves a mitogen-activated protein (MAP) kinase cascade (kinases OS 4, OS-5 and OS-2 downstream of the osmosensing OS-1); the other is less understood, but involves the cut gene, which encodes a putative phosphatase. This study examined turgor regulation, electrical responses, ion fluxes and glycerol accumulation in the cut mutant. Turgor recovery after hyperosmotic treatment was similar to that in the wild-type, for both time-course ( approximately 40 min) and magnitude. Prior to turgor recovery, the hyperosmotic shock caused a rapid transient depolarization of the membrane potential, followed by a sustained hyperpolarization that occurred concomitant with increased H(+) efflux, indicating that the plasma membrane H(+)-ATPase was being activated. These changes also occurred in the wild-type. Net fluxes of Ca(2+) and Cl(-) during turgor recovery were similar to those in the wild-type, but K(+) influx was attenuated in the cut mutant. The similar turgor recovery can be explained by the ion uptake, since glycerol did not accumulate in the cut mutant within the time frame of turgor recovery (but did accumulate in the wild-type). The results suggest that turgor regulation involves multi-faceted coordination of both ion flux and glycerol accumulation. Ion uptake is activated by a MAP kinase cascade, while CUT is required for glycerol accumulation. PMID- 17464063 TI - Use of chromosomal integron arrays as a phylogenetic typing system for Vibrio cholerae pandemic strains. AB - Approximately 200 serogroups of Vibrio cholerae exist, with only two, O1 and O139, responsible for epidemic and pandemic cholera. Strains from these serogroups have evolved from a common progenitor, with lateral gene transfer largely driving their emergence. These strains are so closely related that separation using single- or multi-locus phylogeny has proven difficult. V. cholerae strains contain a genetic system called the integron that is located in the chromosome and that can integrate and excise DNA elements called mobile gene cassettes (MGCs) by site-specific recombination. Large arrays of MGCs are found in V. cholerae strains. For instance, the O1 El Tor strain N16961 contains 179 MGCs. Since integron arrays are dynamic through recombination and excision of MGCs, it was hypothesized that the MGC composition in a given V. cholerae pandemic strain would be useful as a phylogenetic typing system. To address this, a PCR-based method was used to rapidly characterize the MGC composition of V. cholerae arrays. The results showed that the MGC composition of pandemic V. cholerae cassette arrays is relatively conserved, providing further evidence that these strains have evolved from a common progenitor. Comparison of MGC composition between the V. cholerae pandemic strains was also able to resolve the evolution of O139 from a subgroup of O1 El Tor. This level of differentiation of closely related V. cholerae isolates was more sensitive than conventional single gene phylogeny or multi-locus sequence analysis. Using this method, novel MGCs from an O1 classical strain and an Argentinian O139 isolate were also identified, and a major deletion in the MGC array in all pandemic O139 strains and a subset of O1 El Tor strains was identified. Analysis of sequenced V. cholerae integron arrays showed that their evolution can proceed by rearrangements and deletions/insertions of large portions of MGCs in addition to the insertion or excision of single MGCs. PMID- 17464068 TI - Regulation and characterization of rot transcription in Staphylococcus aureus. AB - The pathogenesis of Staphylococcus aureus infections is dependent upon expression of various virulence factors, which are under the control of multiple regulatory systems, including two-component regulatory systems and transcriptional regulators such as the SarA family of proteins. As a part of a continuing effort to understand the regulatory mechanisms that involve the SarA protein family, the regulation and physical characterization of rot transcription is described here. The rot gene, a member of the sarA family of genes, was previously characterized and has been shown to regulate a large number of genes. The rot locus is composed of multiple overlapping transcripts as determined by primer extension and was proposed to encode an open reading frame of 133 residues. Transcription of rot was significantly increased in the sarA mutant. Gel shift and transcriptional studies revealed that SarA could bind to the rot promoter region, probably acting as a repressor for rot transcription. The data indicate that the expression of rot transcription is significantly repressed only by SarA among the sarA family of mutants tested at the post-exponential phase of growth. PMID- 17464069 TI - A two-component small multidrug resistance pump functions as a metabolic valve during nicotine catabolism by Arthrobacter nicotinovorans. AB - The genes nepAB of a small multidrug resistance (SMR) pump were identified as part of the pAO1-encoded nicotine regulon responsible for nicotine catabolism in Arthrobacter nicotinovorans. When [(14)C]nicotine was added to the growth medium the bacteria exported the (14)C-labelled end product of nicotine catabolism, methylamine. In the presence of the proton-motive force inhibitors 2,4 dinitrophenol (DNP), carbonyl cyanide m-chlorophenylhydrazone (CCCP) or the proton ionophore nigericin, export of methylamine was inhibited and radioactivity accumulated inside the bacteria. Efflux of [(14)C]nicotine-derived radioactivity from bacteria was also inhibited in a pmfR : cmx strain with downregulated nepAB expression. Because of low amine oxidase levels in the pmfR : cmx strain, gamma-N methylaminobutyrate, the methylamine precursor, accumulated. Complementation of this strain with the nepAB genes, carried on a plasmid, restored the efflux of nicotine breakdown products. Both NepA and NepB were required for full export activity, indicating that they form a two-component efflux pump. NepAB may function as a metabolic valve by exporting methylamine, the end product of nicotine catabolism, and, in conditions under which it accumulates, the intermediate gamma-N-methylaminobutyrate. PMID- 17464070 TI - Characterization of environmentally friendly nicotine degradation by Pseudomonas putida biotype A strain S16. AB - Nicotine and some related alkaloids in tobacco and tobacco wastes are harmful to health and the environment, and a major environmental requirement is to remove them from tobacco and tobacco wastes. In this study, an isolated strain, S16, identified as Pseudomonas putida biotype A, was used to investigate nicotine degradation. Possible intermediates were identified based on the results of NMR, Fourier-transform (FT)-IR and UV spectroscopy, GC-MS and high-resolution MS (HR MS) analysis. The pathway of nicotine degradation in P. putida was proposed to be from nicotine to 2,5-dihydroxypyridine through the intermediates N methylmyosmine, 2'-hydroxynicotine, pseudooxynicotine, 3-pyridinebutanal,C-oxo, 3 succinoylpyridine and 6-hydroxy-3-succinoylpyridine. N-Methylmyosmine, 2,5 dihydroxypyridine and succinic acid were detected and satisfactorily verified for the first time as intermediates of nicotine degradation. In addition, an alcohol compound, 1-butanone,4-hydroxy-1-(3-pyridinyl), was found to be a novel product of nicotine degradation. These findings provide new insights into the microbial metabolism of nicotine and the environmentally friendly route of nicotine degradation. PMID- 17464071 TI - Structural characterization and ecological roles of a novel exopolysaccharide from the deep-sea psychrotolerant bacterium Pseudoalteromonas sp. SM9913. AB - Pseudoalteromonas sp. SM9913 is a psychrotolerant bacterium isolated from deep sea sediment. The structural characterization and ecological roles of the exopolysaccharide (EPS) secreted by this strain were studied in this work. The yield of the EPS increased as the culture temperature decreased in the range 30 10 degrees C, and it reached 5.25 g l(-1) (dry weight) under optimal growth conditions (15 degrees C, 52 h). EPS fraction was purified and its structure was identified by the combination of NMR spectra, high-resolution mass spectrometry (HRMS) analysis and methylation analysis. The ratio of the sugar units, the acetyl group and the ethoxyl group was close to 4 : 5 : 1. The major sugar unit of the EPS was 6-linked glucose (61.8 %); other sugar units present included terminal arabinofuranosyl (11.0 %) and glucopyranosyl (11.2 %) residues and a small amount of other sugar derivatives. Its structure was different from EPSs reported for other marine bacteria. Besides the structural elucidation of the EPS, its ecological roles were studied. This EPS could enhance the stability of the cold-adapted protease MCP-01 secreted by the same strain through preventing its autolysis. It could bind many metal ions, including Fe(2+), Zn(2+), Cu(2+), Co(2+). It was also a very good flocculating agent and could conglomerate colloidal and suspended particles. These results indicated that the EPS secreted by strain SM9913 might help this strain enrich the proteinaceous particles and the trace metals in the deep-sea environment, stabilize the secreted cold-adapted proteases and avoid its diffusion. This is believed to be the first report on the structure of the EPS secreted by a deep-sea psychrotolerant bacterium and its ecological roles. According to these results and other studies, a schematic diagram of the lifestyle of the deep-sea psychrotolerant strain SM9913 is suggested. PMID- 17464072 TI - Stabilization of water/gas oil emulsions by desulfurizing cells of Gordonia alkanivorans RIPI90A. AB - It has been previously reported that resting-cells, non-proliferating cells, of Gordonia alkanivorans RIPI90A can convert dibenzothiophene (DBT) to 2 hydroxybiphenyl (2-HBP) via the 4S pathway in a biphasic system. The main goal of the current work was to study the behaviour of resting-cells of this strain in biphasic organic media. Resting-cells showed strong affinity for sulfurous organic substrates and were able to stabilize water/gas oil emulsions by attaching to the interface without decreasing the surface tension of their environment. This was consistent with the behaviour of the whole cells but not the surfactants, suggesting that microbial cell-mediated emulsification occurs. It was found that the emulsion-stabilizing activity of the resting-cells was influenced by the growth stage, but was not directly influenced by the metabolic activity of the resting-cells. This activity may be related to cell-surface hydrophobicity, which results from the unique chemical structure of the cell surface. In some biphasic biodesulfurization (BDS) bioreactors, emulsions are created without addition of any surfactant. Cell surface-mediated stabilization helps prolong the emulsions and therefore overcomes mass-transfer limitations in bioreactors. The simultaneous occurrence of emulsion-stabilizing and desulfurization activities of resting-cells was observed for what is believed to be the first time. The results suggest that this strain may have potential for the BDS of diesel oils. PMID- 17464073 TI - Interaction domains in the Pseudomonas aeruginosa type II secretory apparatus component XcpS (GspF). AB - Pseudomonas aeruginosa is an opportunistic pathogen, which secretes a wide variety of enzymes and toxins into the extracellular medium. Most exoproteins are exported by the type II secretion machinery, the Xcp system, which encompasses 12 different proteins. One of the core components of the Xcp system is the inner membrane protein XcpS (GspF), homologues of which can be identified in type II secretion machineries as well as in type IV piliation systems. In this study, XcpS was shown to be stabilized by co-expression of the XcpR (GspE) and XcpY (GspL) components of the machinery, demonstrating an interaction between these three proteins. By replacing segments of P. aeruginosa XcpS with the corresponding parts of its Pseudomonas putida counterpart, XcpS domains were identified that are important for species-specific functioning and thus represent putative interaction domains. The cytoplasmic loop of XcpS was found to be involved in the stabilization by XcpR and XcpY. PMID- 17464074 TI - The outer membrane secretin PilQ from Neisseria meningitidis binds DNA. AB - Neisseria meningitidis is naturally competent for transformation throughout its growth cycle. Transformation in neisserial species is coupled to the expression of type IV pili, which are present on the cell surface as bundled filamentous appendages, and are assembled, extruded and retracted by the pilus biogenesis components. During the initial phase of the transformation process, binding and uptake of DNA takes place with entry through a presumed outer-membrane channel into the periplasm. This study showed that DNA associates only weakly with purified pili, but binds significantly to the PilQ complex isolated directly from meningococcal membranes. By assessing the DNA-binding activity of the native complex PilQ, as well as recombinant truncated PilQ monomers, it was shown that the N-terminal region of PilQ is involved in the interaction with DNA. It was evident that the binding of ssDNA to PilQ had a higher affinity than the binding of dsDNA. The binding of DNA to PilQ did not, however, depend on the presence of the neisserial DNA-uptake sequence. It is suggested that transforming DNA is introduced into the cell through the outer-membrane channel formed by the PilQ complex, and that DNA uptake occurs by non-specific introduction of DNA coupled to pilus retraction, followed by presentation to DNA-binding component(s), including PilQ. PMID- 17464075 TI - svrA, a multi-drug exporter, does not control agr. AB - The Staphylococcus aureus svrA gene was identified in a signature-tagged mutagenesis screen for Tn917 insertions attenuated for mouse virulence, and subsequently found to be defective in agr expression. Its attenuation of virulence was attributed to its failure to express the agr regulon. In addition to the Tn917 insertion in svrA, the original svrA mutant strain (P6C63) has an adventitious frame-shift in agrC, which results in truncation of the AgrC peptide. Separation of the svrA mutation from the agrC frame-shift revealed that svrA has no detectable affect on agr activation, as assessed by exoprotein profiles and the production of haemolytic toxins. These results indicate that svrA does not play a role in Staphylococcus aureus infections via an agr-mediated pathway. PMID- 17464076 TI - Identification of new genes associated with intermediate resistance of Enterococcus faecalis to divercin V41, a pediocin-like bacteriocin. AB - It has been suggested that resistance to class IIa bacteriocins occurs at either a low or a high level. In listerial strains, low-level resistance (2-4-fold) to class IIa bacteriocins is attributed to alterations in membrane lipid composition. In Listeria monocytogenes and Enterococcus faecalis, high-level resistance (1000-fold) correlates with inactivation of the mptACD operon, which encodes the EII(Man)(t) mannose permease of the phosphotransferase system (PTS). Previous studies reported that in L. monocytogenes, high-level resistance involved the sigma(54) factor and the ManR activator. In this investigation, three genes associated with the resistance of Ent. faecalis JH2-2 to divercin V41, a pediocin-like bacteriocin from Carnobacterium divergens V41, were clearly identified by screening an insertional mutant library of Ent. faecalis JH2-2. These genes correspond to the well-known rpoN gene, which encodes sigma(54) factor, and to genes encoding a glycerophosphoryl diester phosphodiesterase (GlpQ) and a protein with a putative phosphodiesterase function (PDE). Resistance of the three mutants defective in the aforementioned genes appeared to be graduated: the rpoN mutant was more resistant than the glpQ mutant, which was more resistant than the pde mutant. Moreover, this resistance was specific to class IIa bacteriocins. PMID- 17464077 TI - Uberolysin: a novel cyclic bacteriocin produced by Streptococcus uberis. AB - Streptococcus uberis is commonly found in the environment and in association with various bovine body sites and is a major cause of bovine mastitis. Moreover, S. uberis is known to produce a variety of bacteriocin-like inhibitory substances, antimicrobial agents that generally inhibit closely related bacterial species. In this respect, S. uberis strain 42 has previously been shown to produce a novel nisin variant named nisin U. This paper reports that, in addition to nisin U, S. uberis strain 42 produces a second bacteriocin that induces the lysis of metabolically active, susceptible target bacteria and which has therefore been named uberolysin. Isolation of the native active antimicrobial agent revealed that uberolysin is a 7048 Da peptide that is refractory to sequence analysis by Edman degradation. Transposon mutagenesis was used to generate a uberolysin negative mutant of S. uberis 42 and sequencing of DNA flanking the insertion site revealed, in addition to the structural gene (ublA), several open reading frames likely to be involved in post-translational modification, transport and producer self-protection (immunity), and possibly in regulation of the biosynthetic gene cluster. In addition, a pair of direct repeats that may be involved in bacteriocin acquisition were identified; indeed, ublA could be identified in 18 % of tested S. uberis strains. Enzymic hydrolysis of uberolysin was used to confirm that ublA does indeed encode the precursor of uberolysin, that an unusually short leader sequence of only six amino acids is cleaved during processing of the mature peptide and that uberolysin is post-translationally covalently modified to form a head-to-tail monocycle. Thus, uberolysin is a unique cyclic bacteriocin, belonging to the same family of bacteriocins as enterocin AS-48 and circularin A. PMID- 17464078 TI - Isolation and partial characterization of the Streptococcus mutans type AII lantibiotic mutacin K8. AB - Streptococcus mutans strain K8 was shown to produce a newly identified type AII lantibiotic, mutacin K8. The mutacin K8-encoding muk locus consists of 13 ORFs, three of which (mukA1, A2 and A3) have close homology to scnA, the structural gene encoding the Streptococcus pyogenes lantibiotic SA-FF22, and another (mukA') resembles scnA', an ORF in the SA-FF22 locus that has no currently assigned function. Inactivation of the muk locus indicated that mutacin K8 is responsible for most of the inhibitory activity produced by strain K8 in deferred antagonism tests on Columbia blood agar base supplemented with 5 % human blood and 0.1 % CaCO(3). By contrast, on tryptic soy agar plus 2 % yeast extract and 0.5 % CaCO(3) most of the inhibitory activity of strain K8 appeared to be attributable either to mutacin IV or to some other inhibitory peptide(s) exported by the mutacin IV transporter nlmT. An inhibitory peptide purified from a derivative of strain K8 in which nlmT had been inactivated had a mass of 2734 Da and an N terminal sequence identical to the predicted propeptide translation products of mukA1 and mukA3. The muk locus may be widely distributed in S. mutans, since 9 (35 %) of 26 strains tested contained at least part of the locus. In the genome sequence of strain UA159 the muk locus is incomplete, the sole residual components being the ORFs encoding the putative two-component regulatory system mukR (SMU.1815) and mukK (SMU.1814), followed by two transposases (SMU.1813 and SMU.1812) and then the ORFs mukF (SMU.1811), mukE (SMU.1810) and mukG (SMU.1809), thought to encode putative immunity peptides. Strains such as UA159 having incomplete loci did not produce detectable levels of mutacin K8. PMID- 17464079 TI - Identification and functional analysis of a phytoene desaturase gene from the extremely radioresistant bacterium Deinococcus radiodurans. AB - The phytoene-related desaturases are the key enzymes in the carotenoid biosynthetic pathway. The gene encoding phytoene desaturase in the deinoxanthin synthesis pathway of Deinococcus radiodurans was identified and characterized. Two putative phytoene desaturase homologues (DR0861 and DR0810) were identified by analysis of conserved amino acid regions, and the former displayed the highest identity (68 %) with phytoene desaturase of the cyanobacterium Gloeobacter violaceus. DR0861 gene knockout and dinucleotide-binding motif deletion resulted in the arrest of lycopene synthesis and the accumulation of phytoene. The colourless DR0861 knockout mutant became more sensitive to acute ionizing radiation and oxygen stress. Complementation of the mutant with a heterologous or homologous gene restored its pigment and resistance. The desaturase activity of DR0861 (crtI) was further confirmed by the assay of enzyme activity in vitro and heterologous expression in Escherichia coli containing crtE and crtB genes (responsible for phytoene synthesis) from Erwinia uredovora. In addition, the amount of lycopene synthesis in E. coli resulting from the expression of crtI from D. radiodurans was determined, and this had significant dose-dependent effects on the survival rate of E. coli exposed to hydrogen peroxide and ionizing radiation. PMID- 17464080 TI - Modulation of lambda plasmid and phage DNA replication by Escherichia coli SeqA protein. AB - SeqA protein, a main negative regulator of the replication initiation of the Escherichia coli chromosome, also has several other functions which are still poorly understood. It was demonstrated previously that in seqA mutants the copy number of another replicon, the lambda plasmid, is decreased, and that the activity of the lambda p(R) promoter (whose function is required for stimulation of ori lambda) is lower than that in the wild-type host. Here, SeqA-mediated regulation of lambda phage and plasmid replicons was investigated in more detail. No significant influence of SeqA on ori lambda-dependent DNA replication in vitro was observed, indicating that a direct regulation of lambda DNA replication by this protein is unlikely. On the other hand, density-shift experiments, in which the fate of labelled lambda DNA was monitored after phage infection of host cells, strongly suggested the early appearance of sigma replication intermediates and preferential rolling-circle replication of phage DNA in seqA mutants. The directionality of lambda plasmid replication in such mutants was, however, only slightly affected. The stability of the heritable lambda replication complex was decreased in the seqA mutant relative to the wild-type host, but a stable fraction of the lambda O protein was easily detectable, indicating that such a heritable complex can function in the mutant. To investigate the influence of seqA gene function on heritable complex- and transcription-dependent lambda DNA replication, the efficiency of lambda plasmid replication in amino acid-starved relA seqA mutants was measured. Under these conditions, seqA dysfunction resulted in impairment of lambda plasmid replication. These results indicate that unlike oriC, SeqA modulates lambda DNA replication indirectly, most probably by influencing the stability of the lambda replication complex and the transcriptional activation of ori lambda. PMID- 17464081 TI - Sequence analysis of the mobilizable lactococcal plasmid pGdh442 encoding glutamate dehydrogenase activity. AB - A novel plasmid named pGdh442 had previously been isolated from a plant Lactococcus lactis strain. This plasmid encodes two interesting properties with applications in the dairy industry: a glutamate dehydrogenase activity that stimulates amino acid conversion to aroma compounds, and cadmium/zinc resistance that can be used as a selectable marker. Moreover, this plasmid can be transferred naturally to other strains, but appears to be incompatible with certain other lactococcal plasmids. During this study, the complete sequence of pGdh442 (68 319 bp) was determined and analysed. This plasmid contains 67 ORFs that include 20 IS elements that may have mediated transfer events between L. lactis and other genera living in the same biotope, such as Streptococcus, Pediococcus and Lactobacillus. Even though it is a low-copy-number plasmid, it is relatively stable due to a theta replication mode and the presence of two genes involved in its maintenance system. However, pGdh442 is incompatible with pSK08 derived protease/lactose plasmids because both possess the same replication and partition system. pGdh442 is not self-transmissible, but can be naturally transmitted via mobilization by conjugative elements carried by the chromosome or by other plasmids, such as the 712-type sex factor, which is widely distributed in L. lactis. In addition to several genes already found on other L. lactis plasmids, such as the oligopeptide transport and utilization genes, pGdh442 also carries several genes not yet identified in L. lactis. Finally, it does not carry genes that would trigger concern over its presence in human food. PMID- 17464082 TI - Erythropoietin in cancer: presumption of innocence? AB - Erythropoietin emerged as the biggest drug in oncology despite never having demonstrated a survival benefit in patients with cancer. Two phase III clinical trials reported more than 3 years ago that erythropoietin adversely affected cancer survival rates, due mainly to tumor progression. Despite changes to the product label for erythropoietins in 2004, clinical practice remained unchanged until recent weeks when, following reports of three new phase III studies and a phase II trial, a "black box warning" for erythropoietin products was issued by the Food and Drug Administration (FDA). Whether erythropoietin products can be considered safe when used for FDA-approved indications is currently at issue; however, addressing this question will be difficult until the mechanisms of erythropoietin-stimulated tumor progression are understood. A thorough evaluation of materials from clinical trials already completed may shed new light on how erythropoietin promotes cancer progression. Until these issues are resolved, oncologists should inform their patients of erythropoietin's potential adverse impact on cancer progression and survival. Disclosure of potential conflicts of interest is found at the end of this article. PMID- 17464083 TI - Circulating connective tissue precursors: extreme rarity in humans and chondrogenic potential in guinea pigs. AB - Using a variety of cell separation techniques and cultivation conditions, circulating, adherent, connective tissue, clonogenic cells were found in just 3 donors out of 66, demonstrating that these precursors are extremely rare in postnatal human blood. Contrary to humans, guinea pig blood shows much more reproducible connective tissue colony formation; it was therefore chosen to study the differentiation potential of adherent blood-derived clonogenic cells. Out of 22 single colony-derived strains of various morphologies, only 5 spindle-shaped strains showed extensive proliferative capacity in vitro. None of these strains formed bone upon in vivo transplantation, whereas two strains formed cartilage in high-density pellet cultures in vitro. Both chondrogenic strains included cells expressing aggrecan, whereas nonchondrogenic strains did not. Out of four polyclonal strains studied, one formed both cartilage and abundant bone accompanied by hematopoiesis-supporting stroma. Evidently, there are cells in adult guinea pig blood capable of both extensive proliferation and differentiation toward cartilage: circulating chondrogenic precursors. Although some of these cells lack osteogenic potential and therefore represent committed chondrogenic precursors, others may be multipotential and consequently belong to the family of skeletal stem cells. This is the first demonstration of postnatal circulating chondrogenic precursors, as well as of precursor cells with chondrogenic but not osteogenic potential. Disclosure of potential conflicts of interest is found at the end of this article. PMID- 17464084 TI - Clonal analysis of human embryonic stem cell differentiation into teratomas. AB - Differentiation of human embryonic stem cells (HESCs) can be studied in vivo through the induction of teratomas in immune-deficient mice. Cells within the teratomas differentiate into all three embryonic germ layers. However, the exact nature of the proliferation and differentiation of HESCs within the teratoma is not fully characterized, and it is not clear whether the differentiation is cell autonomous or affected by neighboring cells. Here, we establish a genetic approach to study the clonality of differentiation in teratomas using a mixture of HESC lines. We first demonstrate, by means of 5-bromo-2'-deoxyuridine incorporation, that cell proliferation occurs throughout the teratoma, and that there are no clusters of undifferentiated-proliferating cells. Using a combination of laser capture microdissection and DNA fingerprinting analysis, we show that different cell lines contribute mutually to the same distinctive tissue structures. Further support for the nonclonal differentiation within the teratoma was achieved by fluorescence in situ hybridization analysis of sex chromosomes. We therefore suggest that in vivo differentiation of HESCs is polyclonal and, thus, may not be cell autonomous, stressing the need for a three-dimensional growth in order to achieve complex differentiation of HESCs. Disclosure of potential conflicts of interest is found at the end of this article. PMID- 17464085 TI - A novel function of interleukin-10 promoting self-renewal of hematopoietic stem cells. AB - Self-renewal of hematopoietic stem cells (HSCs) is key to their reconstituting ability, but the factors regulating the process remain poorly understood. Here, we show that Interleukin-10 (IL-10), a pleiotropic immune modulating cytokine, can also play a role in regulating HSC self-renewal. First, a quantitative decrease of primitive hematopoietic cell populations, but not more matured cells, was observed in the bone marrows of IL-10 disrupted mice as determined by long term in vitro cultures or in vivo competitive repopulation assays. In contrast, normal HSCs from 5-fluorouracil treated marrows cultured on the IL-10 secreting stroma displayed an enhanced repopulating activity compared with cells grown on control stroma, with ninefold higher numbers of donor-derived HSCs in the reconstituted recipient marrows. Moreover, limiting dilution transplantation assay demonstrated that exogenous addition of IL-10 in the stroma-free cultures of purified Lin- Sca-1+ c-kit+ cells caused three- to fourfold higher frequencies of HSCs in the 5-day short-term culture without indirect inhibitory effect of IL 10 on tumor necrosis factor-alpha or interferon-gamma secretion. Interestingly, primitive hematopoietic cells, including Lin- Sca-1+ c-kit+ or side population cells, expressed the surface receptor for IL-10, and microenvironmental production of IL-10 was sharply increased in the osteoblasts lining the trabecular regions of the radiation-stressed marrow but not in the steady-state marrows. These results show that IL-10 may be a ligand that can stimulate self renewal of HSCs to promote their regeneration in addition to being a ligand for immune regulation. Disclosure of potential conflicts of interest is found at the end of this article. PMID- 17464086 TI - Contribution of bone marrow-derived stem cells to endometrium and endometriosis. AB - Bone marrow-derived cells (BMDCs) can differentiate into nonhematopoietic cells, suggesting that BMDCs may contribute to the maintenance of multiple tissues. Donor-derived bone marrow cells have been identified in human uterine endometrium. Here, two murine models were used to investigate the contribution of nonendometrial stem cells to endometrium. We investigate whether BMDCs can localize to uterine endometrium and to endometriosis. After bone marrow transplantation, male donor-derived bone marrow cells were found in the uterine endometrium of female mice. Although uncommon (<0.01%), these cells can differentiate into epithelial cells. After generation of experimental endometriosis by ectopic endometrial implantation in the peritoneal cavity, bone marrow from LacZ transgenic mice was used for transplantation. LacZ expressing cells were found in the wild-type ectopic endometrium implanted in the peritoneal cavity of hysterectomized LacZ transgenic mice. The repopulation of endometrium with bone marrow-derived stem cells may be important to normal endometrial physiology and also may help to explain the cellular basis for the high long-term failure of conservative alternatives to hysterectomy. The examination of a sexually dimorphic organ such as the uterus demonstrates the ability of male bone marrow, which cannot harbor circulating endometrial cells, to generate endometrium de novo and proves their mesenchymal stem cell origin. Finding Y chromosome bearing endometrial cells demonstrates the potential to recapitulate embryonic developmental pathways that were never activated in males; BMDCs may have vast regenerative capacity. Additionally, the ability of stem cells to engraft endometriosis has implications for the origin and progression of this disease. Ectopic differentiation of stem cells may be a novel mechanism of disease. Disclosure of potential conflicts of interest is found at the end of this article. PMID- 17464087 TI - Complete spinal cord injury treatment using autologous bone marrow cell transplantation and bone marrow stimulation with granulocyte macrophage-colony stimulating factor: Phase I/II clinical trial. AB - To assess the safety and therapeutic efficacy of autologous human bone marrow cell (BMC) transplantation and the administration of granulocyte macrophage colony stimulating factor (GM-CSF), a phase I/II open-label and nonrandomized study was conducted on 35 complete spinal cord injury patients. The BMCs were transplanted by injection into the surrounding area of the spinal cord injury site within 14 injury days (n = 17), between 14 days and 8 weeks (n = 6), and at more than 8 weeks (n = 12) after injury. In the control group, all patients (n = 13) were treated only with conventional decompression and fusion surgery without BMC transplantation. The patients underwent preoperative and follow-up neurological assessment using the American Spinal Injury Association Impairment Scale (AIS), electrophysiological monitoring, and magnetic resonance imaging (MRI). The mean follow-up period was 10.4 months after injury. At 4 months, the MRI analysis showed the enlargement of spinal cords and the small enhancement of the cell implantation sites, which were not any adverse lesions such as malignant transformation, hemorrhage, new cysts, or infections. Furthermore, the BMC transplantation and GM-CSF administration were not associated with any serious adverse clinical events increasing morbidities. The AIS grade increased in 30.4% of the acute and subacute treated patients (AIS A to B or C), whereas no significant improvement was observed in the chronic treatment group. Increasing neuropathic pain during the treatment and tumor formation at the site of transplantation are still remaining to be investigated. Long-term and large scale multicenter clinical study is required to determine its precise therapeutic effect. Disclosure of potential conflicts of interest is found at the end of this article. PMID- 17464088 TI - Simvastatin suppresses self-renewal of mouse embryonic stem cells by inhibiting RhoA geranylgeranylation. AB - Statins, 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors, were originally developed to lower cholesterol. Their pleiotropic (or cholesterol independent) effects at the cellular and molecular levels are highly related to numerous cellular functions, such as proliferation and differentiation. However, they are hardly studied in embryonic stem cells. In this study, we evaluated the effects of statins on mouse ESCs (J1, D3, and RW.4) to enhance our understanding of the molecular basis of ESC self-renewal. Treatment of ESCs with simvastatin, mevastatin, atorvastatin, or pravastatin induced morphological change and decreased cell proliferation. We observed that the use of simvastatin was most effective in all three ESCs. Loss of ESC self-renewal by simvastatin was determined by marked downregulation of ESC markers alkaline phosphatase, Oct4, Nanog, Rex-1, and SSEA-1. Simvastatin effects were selectively reversed by either mevalonate or its metabolite geranylgeranyl pyrophosphate (GGPP) but not by cholesterol or farnesyl pyrophosphate. These results suggest that simvastatin effects were mainly derived from depletion of intracellular pools of GGPP, the substrate required for the geranylgeranylation. Using this approach, we found that GGPP, a derivative of the mevalonate pathway, is critical for ESC self renewal. Furthermore, we identified that simvastatin selectively blocked cytosol to-membrane translocalization of RhoA small guanosine triphosphate-binding protein, known to be the major target for geranylgeranylation, and lowered the levels of Rho-kinase (ROCK)2 protein in ESCs. In addition, simvastatin downregulated the ROCK activity, and this effect was reversed by addition of GGPP. Our data suggest that simvastatin, independently of its cholesterol lowering properties, impairs the ESC self-renewal by modulating RhoA/ROCK dependent cell-signaling. PMID- 17464089 TI - Sphingosine 1-phosphate mediates proliferation and survival of mesoangioblasts. AB - Mesoangioblasts are stem cells capable of differentiating in various mesodermal tissues and are presently regarded as suitable candidates for cell therapy of muscle degenerative diseases, as well as myocardial infarction. The enhancement of their proliferation and survival after injection in vivo could greatly improve their ability to repopulate damaged tissues. In this study, we show that the bioactive sphingolipid sphingosine 1-phosphate (S1P) regulates critical functions of mesoangioblast cell biology. S1P evoked a full mitogenic response in mesoangioblasts, measured by labeled thymidine incorporation and cell counting. Moreover, S1P strongly counteracted the apoptotic process triggered by stimuli as diverse as serum deprivation, C2-ceramide treatment, or staurosporine treatment, as assessed by cell counting, as well as histone-associated fragments and caspase 3 activity determinations. S1P acts both as an intracellular messenger and through specific membrane receptors. Real-time polymerase chain reaction analysis revealed that mesoangioblasts express the S1P-specific receptor S1P3 and, to a minor extent, S1P1 and S1P2. By using S1P receptor subtype-specific agonists and antagonists, we found that the proliferative response to S1P was mediated mainly by S1P2. By contrast, the antiapoptotic effect did not implicate S1P receptors. These findings demonstrate an important role of S1P in mesoangioblast proliferation and survival and indicate that targeting modulation of S1P dependent signaling pathways may be used to improve the efficiency of muscle repair by these cells. Disclosure of potential conflicts of interest is found at the end of this article. PMID- 17464090 TI - The late dividing population of gamma-retroviral vector transduced human mobilized peripheral blood progenitor cells contributes most to gene-marked cell engraftment in nonobese diabetic/severe combined immunodeficient mice. AB - We used the nonobese diabetic/severe combined immunodeficient (NOD/SCID) mouse model to assess the repopulation potential of subpopulations of mobilized human CD34+ peripheral blood progenitor cells (PBPC). First, PBPC were transduced with gamma-retrovirus vector RD114-MFGS-CFP, which requires cell division for successful transduction, at 24 hours, 48 hours, and 72 hours to achieve 96% cyan fluorescent protein (CFP)-positive cells. Cells were sorted 12 hours after the last transduction into CFP-positive (divided cells) and CFP-negative populations. CFP-positive cells were transplanted postsort, whereas the CFP-negative cells were retransduced and injected at 120 hours. The CFP-negative sorted and retransduced cells contained markedly fewer vector copies and resulted in a 32 fold higher overall engraftment and in a 13-fold higher number of engrafted transgene positive cells. To assess cell proliferation as an underlying cause for the different engraftment levels, carboxyfluorescein succinimidyl ester-labeling of untransduced PBPC was performed to track the number of cell divisions. At 72 hours after initiation of culture, when 95% of all cells have divided, PBPC were sorted into nondivided and divided fractions and transplanted into NOD/SCID mice. Nondivided cells demonstrated 45-fold higher engraftment than divided cells. Late dividing PBPC in ex vivo culture retain high expression of the stem cell marker CD133, whereas rapidly proliferating cells lose CD133 in correlation to the number of cell divisions. Our studies demonstrate that late dividing progenitors transduced with gamma-retroviral vectors contribute most to NOD/SCID engraftment and transgene marking. Confining the gamma-retroviral transduction to CD133 positive cells on days 3 and 4 could greatly reduce the number of transplanted vector copies, limiting the risk of leukemia from insertional mutagenesis. Disclosure of potential conflicts of interest is found at the end of this article. PMID- 17464091 TI - Modeling notch signaling in normal and neoplastic hematopoiesis: global gene expression profiling in response to activated notch expression. AB - In normal hematopoiesis, proliferation is tightly linked to differentiation in ways that involve cell-cell interaction with stromal elements in the bone marrow stem cell niche. Numerous in vitro and in vivo studies strongly support a role for Notch signaling in the regulation of stem cell renewal and hematopoiesis. Not surprisingly, mutations in the Notch gene have been linked to a number of types of malignancies. To better define the function of Notch in both normal and neoplastic hematopoiesis, a tetracycline-inducible system regulating expression of a ligand-independent, constitutively active form of Notch1 was introduced into murine E14Tg2a embryonic stem cells. During coculture, OP9 stromal cells induce the embryonic stem cells to differentiate first to hemangioblasts and subsequently to hematopoietic stem cells. Our studies indicate that activation of Notch signaling in flk+ hemangioblasts dramatically reduces their survival and proliferative capacity and lowers the levels of hematopoietic stem cell markers CD34 and c-Kit and the myeloid marker CD11b. Global gene expression profiling of day 8 hematopoietic progenitors in the absence and presence of activated Notch yield candidate genes required for normal hematopoietic differentiation, as well as putative downstream targets of oncogenic forms of Notch including the noncanonical Wnts Wnt4 and 5A. Disclosure of potential conflicts of interest is found at the end of this article. PMID- 17464092 TI - Essential alterations of heparan sulfate during the differentiation of embryonic stem cells to Sox1-enhanced green fluorescent protein-expressing neural progenitor cells. AB - Embryonic stem (ES) cells can be cultured in conditions that either maintain pluripotency or allow differentiation to the three embryonic germ layers. Heparan sulfate (HS), a highly polymorphic glycosaminoglycan, is a critical cell surface coreceptor in embryogenesis, and in this paper we describe its structural transition from an unusually low-sulfated variant in ES cells to a more highly sulfated form in fluorescence-activated cell sorting-purified neural progenitor cells. The characteristic domain structure of HS was retained during this transformation. However, qualitative variations in surface sulfation patterns between ES and differentiated cells were revealed using HS epitope-specific antibodies and the HS-binding growth factor fibroblast growth factor 2 (FGF-2). Expression profiles of the HS modification enzymes indicated that both "early" (N sulfotransferases) and "late" (6O- and 3O-sulfotransferases) sulfotransferases contributed to the alterations in sulfation patterning. An HS-null ES line was used to demonstrate the necessity for HS in neural differentiation. HS is a coreceptor for many of the protein effectors implicated in pluripotency and differentiation (e.g., members of the FGF family, bone morphogenic proteins, and fibronectin). We suggest that the stage-specific activities of these proteins are finely regulated by dynamic changes in sulfation motifs in HS chains. Disclosure of potential conflicts of interest is found at the end of this article. PMID- 17464093 TI - Homocysteine, folate and cobalamin levels in hypothyroid women before and after treatment. AB - Hypothyroidism may result in accelerated atherosclerosis. Hyperhomocysteinaemia is an independent risk factor for premature atherosclerotic vascular disease. The aim of the present study was to assess plasma total homocysteine (tHcy), folate and cobalamin concentrations in hypothyroid patients before and after treatment. Thirty-one hypothyroid and thirty health young women were studied. The hypothyroid patients were investigated in the untreated state and again after restoration of euthyroidism. The levels of homocysteine, folate, cobalamin and thyroid stimulating hormone (TSH), free thyroxine (fT(4)), free triiodothyronine (fT(3)) and renal function were measured before and after treatment. In hypothyroidism tHcy was higher but not statistically significant than in control group. Serum level of folate was higher and serum cobalamin was lower in the hypothyroid state. Following L-thyroxine therapy tHcy significantly decreased as well as the concentration of cobalamin. Level of folate remained unchanged. Univariate analysis in hypothyroid group indicated that tHcy negative correlated with creatinine clearance, fT(3), fT(4), cobalamin and positive with TSH. In multivariate analysis tHcy correlated with creatinine clearance, cobalamin and fT(4). Thyroid status influences the plasma tHcy. Free triiodothyronine and next free thyroxine have the greatest negative influence. This would account for hyperhomocysteinemia in the hypothyroid state and premature atherogenesis. PMID- 17464094 TI - Fibroblast growth factor-23 (FGF23) in patients with transient hypoparathyroidism: its important role in serum phosphate regulation. AB - Hypoparathyroidism is a complication of thyroidectomy that causes hyperphosphatemia primarily due to enhanced reabsorption of phosphate in the kidney resulting from decreased parathyroid hormone (PTH) secretion. Fibroblast growth factor-23 (FGF23) is a hormone-like factor that is thought to play an important role in phosphate homeostasis. However, the changes and role of FGF23 in transient hypoparathyroidism after thyroidectomy are not clear. We examined changes in serum levels of calcium, phosphate, intact PTH, 1,25-dihydroxyvitamin D, and FGF23 in 12 patients (10 women, 2 men; mean age, 51 yr) who developed transient hypoparathyroidism after thyroidectomy. Serum phosphate reached its peak level (5.9 +/- 0.5 mg/dl) approximately 4 days after development of hypoparathyroidism, and this was followed by a peak in the serum FGF23 level (71 +/- 28 ng/l). Serum levels of calcium, phosphate, and FGF23 normalized after recovery of parathyroid function. There was a significant positive correlation between serum phosphate and FGF23 levels (P<0.05). Serum FGF23 was elevated in patients with hypoparathyroidism and hyperphosphatemia and normalized along with normalized phosphate levels after recovery of parathyroid function. The peak level of phosphate always preceded that of FGF23 by several days, suggesting that elevated phosphate is a primary stimulus for release of FGF23. This homeostatic regulation of phosphate differs considerably from that of serum calcium whose change is rapidly corrected within minutes. PMID- 17464095 TI - Effect of curcumin and curcumin copper complex (1:1) on radiation-induced changes of anti-oxidant enzymes levels in the livers of Swiss albino mice. AB - The effect of mononuclear copper (II) complex of curcumin in 1:1 stoichiometry (hereafter referred to as complex) administered 30 min before gamma-irradiation (4.5 Gy) on alterations in antioxidant and Thiobarbituric acid reactive substances (TBARS) levels in livers was studied in comparison to curcumin at a dose of 50 mg/kg. The different antioxidants like GSH, GST, catalase, SOD, TBARS and total thiols were estimated in the liver homogenates excised at different time intervals (1, 2 and 4 h) post irradiation using colorimetric methods. There was a radiation-induced decrease in the levels of all the studied enzymes at 1 h post irradiation, while an increase was observed at later time points. Both curcumin and complex treatment in sham-irradiated mice decreased the levels of GSH and total thiols, whereas there was an increase in the levels of catalase, GST and SOD compared to normal control. Under the influence of irradiation, both curcumin and complex treatment protected the decline in the levels of GSH, GST, SOD, catalase and total thiols, and inhibited radiation-induced lipid peroxidation. Further, the complex was found to be more effective in protecting the enzymes at 1 h post irradiation compared to curcumin treated group. This may be due to the higher rate constants of the complex compared to curcumin for their reactions with various free radicals. PMID- 17464096 TI - Role of enhanced radiosensitivity and the tumor-specific suicide gene vector in gene therapy of nasopharyngeal carcinoma. AB - Nasopharyngeal carcinoma (NPC) is one of the common malignant tumors in China. Radiotherapy and chemotherapy are the main therapy methods for NPC. To enhance the specific antitumor effect, a novel vector with radiosensitivity and tumor specificity was constructed in this study, which enables the reduction of dosage of radiation and chemotherapeutic drugs due to its double killing effect. Four DNA elements, Egr-1 promoter, Cytosine deaminase (CD) gene, hTERT promoter, Survivin antisense oligonucleotides were amplified and constructed in pcDNA3.1 vector. CD and Survivin gene expression in CNE-2 cells were detected by RT-PCR. High performance liquid chromatography (HPLC) was employed to determine the transformation from the prodrugs 5-FC to 5-FU. Hoechst33258 staining of the nuclei and methylthiazolyl tetrazolium(MTT) assay were applied to detect apoptosis and cell survivability, respectively. In addition, the anti-tumor effects were examined in vivo by injecting cells with different vectors into nude mice. Our results revealed a notable killing effect of combined treatment with 5 FC and radiation on CNE-2 cells transfected with vectors in vitro. This effect was especially notable on pEC-TS transferred cells, which showed 57% of cells were killed. In vivo, an obvious suppression of tumor was displayed in pEC-TS group, which was significantly different from other groups (p < 0.05). Consequently, this expression cassette may have a great therapeutic potential for the treatment of NPC. PMID- 17464097 TI - Hormone replacement therapy and cancers: the biological roles of estrogen and progestin in tumorigenesis are different between the endometrium and breast. AB - Hormone replacement therapy (HRT) has become available over the past few decades, but the risk of breast cancer with HRT remains controversial. The Women's Health Initiative Study has recently demonstrated that women receiving estrogen plus progestin (HRT) have an increased risk of invasive breast carcinoma, although women receiving estrogen alone (estrogen replacement therapy) exhibit no increased risk of breast carcinoma. By contrast, the risk of endometrial carcinoma increases with estrogen replacement therapy, while HRT reduces the risk of endometrial carcinoma. These clinical findings suggest that the biological roles of estrogen and progestin in tumorigenesis are certainly different between the endometrium and breast, although both are considered "estrogen-dependent tissues". In this review, I summarize the recent studies and indicate that the enzymes responsible for intratumoral estrogen metabolism and biosynthesis are markedly different between human breast and endometrial carcinomas. 17beta hydroxysteroid dehydrogenases (17-HSDs) are enzymes estrogen replacement therapyinvolved in the formation of active sex steroids. Estrogens are interconverted by two enzymes, 17-HSD types 1 and 2. Type 1 converts estrone to estradiol, and type 2 catalyzes the reverse reaction. 17-HSD type 5 reduces androstenedione to testosterone. 17-HSD type 1 plays an important role in the regulation of high estradiol levels in breast carcinoma tissues, whereas 17-HSD types 2 and 5 appear to be essential for the maintenance of estradiol concentrations in endometrial carcinoma tissues. In addition, the biological significance of progesterone receptor isoforms differs between endometrial and breast carcinomas. These findings may provide new insights into the biology of "estrogen-dependent tissues". PMID- 17464098 TI - Proper assessment of the falling risk in the elderly by a physical mobility test with an obstacle. AB - The Timed "Up & Go" (TUG) is a representative mobility test for assessing the falling risk of the elderly. Although several tests have been developed, including the TUG, these do not include a "tripping" element, and tripping is a major cause of falling. This study examined the influence of various obstacle heights on test performance in the TUG test and test-retest reliability. Twenty two healthy elderly women participated in the TUG test and in the TUG test with an obstacle (TUGO). The obstacle is a box (width 120 cm and depth 20 cm), the height of which varies (0, 5 and 17 cm). In the 0-cm height condition, a thin sheet was laid down instead of the box. In the TUGO, subjects stood up from an armchair, walked 5 m, stepped over the box, turned, stepped over the box again, walked back to the chair and sat down. The reliability of the time required for the motion around the obstacle was high and the total time in the TUGO test increased (intra-class correlation coefficient = 0.74-0.99). These parameters were significantly larger for the height order 17, 5, and 0 cm and exhibited significant correlations with time required for the TUG (r = 0.61 - 0.92) and the height of subjects. An obstacle with 5 cm height prolonged the time during standing on one leg just before the obstacle and the time during turning motion after stepping over it. By adding the obstacle to the TUG, the physical mobility of the elderly is assessed more properly. PMID- 17464099 TI - Long-term cigarette smoking increases the prevalence of carotid artery calcification seen on panoramic dental radiographs in male patients. AB - Panoramic dental radiographs are commonly used in general dentistry and oral and maxillofacial surgery to examine upper and lower teeth, maxilla, mandible and the surroundings simultaneously. Carotid artery calcification, a specific indicator of atherosclerotic change of the carotid arteries, can be seen on the radiographs. Many studies have suggested that cigarette smoking is a risk factor of atherosclerotic change as well as cerebral infarction. We hypothesized that smoking could increase the prevalence of carotid artery calcification, and compared the radiographs of smokers and non-smokers aged 50 years and over: 146 male smokers, 165 male non-smokers, 42 female smokers and 422 female non-smokers. This is the first study to focus on carotid artery calcification seen on panoramic dental radiographs to show the connection between smoking and atherosclerotic change. In male patients, carotid artery calcification was seen in 18 (14.1%) of the smokers, and in 8 (4.8%) of the non-smokers, which clearly shows that male patients aged 50 years old or over are more likely to develop carotid artery calcification if they smoke. However, there is no significant difference between female smokers and female non-smokers in the same age group. Dentists are in a good position to find carotid artery calcification on radiographs. When this is found on a radiograph, the patient should be advised to stop smoking and be referred to a physician for further tests. Clinicians should be aware that this radiographic finding indicates the presence of atherosclerotic change of the carotid arteries. PMID- 17464100 TI - Genetic variations within the insulin gene region are associated with accelerated fetal growth. AB - Size at birth has been proposed to be associated with the risk of type 2 diabetes and cardiovascular disease later in life. It is, however, unclear whether this association is attributed to an unfavorable intrauterine environment or to specific genotypes predisposing both altered fetal growth and common diseases in adult life. The aim of this study was to investigate the associations between the neonatal birth size and the genotypes of polymorphic loci within the insulin gene (INS) region, which is susceptible to diabetes mellitus. We analyzed the genotypes of two polymorphic loci; -23HphI and HUMTH01, in 520 pairs of normal Japanese mothers and their neonates, and compared with the somatoscopic characteristics at birth converted into standard deviation scores (SDS) according to sex, parity and gestational weeks at delivery. It was revealed that neonatal 23HphI T allele and HUMTH01 allele10, which are linked to the INS variable number of tandem repeats (VNTR) class III allele, were associated with increased weight, head circumstance, and length at birth. These associations confirmed that variation within the INS region, most probably at the INS-VNTR, influences fetal growth. Furthermore, the finding that the paternally transmitted -23HphI T allele was exclusively correlated with increased size at birth indicates the involvement of an imprinting mechanism. In conclusion, the INS-VNTR class III allele might accelerate fetal growth in a parent-specific manner. PMID- 17464101 TI - Correlation between clinicopathology and expression of heat shock protein 72 and glycoprotein 96 in human gastric adenocarcinoma. AB - Heat shock protein 72 (HSP72) and glycoprotein 96 (gp96) are highly expressed in cancer tissues. Recent studies indicate the possible roles of HSP72 and gp96 in the development and progression of gastric carcinomas but detailed information is still ambiguous. In this study, we investigated the correlation between clinicopathology and expression of HSP72 and gp96 in human gastric carcinoma. The expression of HSP72 and gp96 was studied in 60 human gastric carcinomas with or without metastasis as well as in mucous membrane adjacent to cancers by way of immunohistochemistry. HSP72 immunoreactivities were detected in 54 of 60 primary tumors (90.0%) and in 22 of 60 mucous membranes adjacent to cancers (36.7%). Likewise, gp96 immunoreactivities were detected in 49 cases of gastric carcinoma (81.7%) and in 15 samples of mucous membrane adjacent to cancer (25.0%). Both HSP72 and gp96 were stained in cytoplasm. HSP72 and gp96 expression in colonic carcinomas with metastasis was significantly higher than those with non metastasis (p < 0.05). The results indicate that there exists a significant correlation between the expression of HSP72 and gp96 and the progression of gastric carcinomas. The high-level expression of HSP72 and gp96 may be used as diagnostic or prognostic markers for gastric carcinoma. PMID- 17464102 TI - Pistachio intake increases high density lipoprotein levels and inhibits low density lipoprotein oxidation in rats. AB - There is increasing evidence that nuts have protective effects against coronary artery disease by improving lipid profile and inhibiting lipid oxidation. However, data about pistachio nuts are limited, and to our knowledge, there is no study investigating the effects of pistachio intake on lipid oxidation and serum antioxidant levels. This study, therefore, sought to determine the effects of pistachio intake on serum lipids and determine whether consumption of pistachio would alter serum antioxidant levels. Rats were randomly divided into three groups (n=12 for each): control group fed basic diet for 10 weeks and treated groups fed basic diet plus pistachio which constituted 20% and 40% of daily caloric intake, respectively. Consumption of pistachio as 20% of daily caloric intake increased high-density lipoprotein (HDL) levels and decreased total cholesterol (TC)/HDL ratio, compared with those not taking pistachio. However, TC, low-density lipoprotein (LDL) cholesterol and triglyceride levels were unaffected by pistachio consumption. Consumption of pistachio as 20% of daily caloric intake increased serum paraoxonase activity by 35% and arylesterase activity by 60%, which are known to inhibit LDL cholesterol oxidation, compared with the control group. However, increased antioxidant activity was blunted when pistachio intake was increased to 40% of daily caloric intake. In conclusion, the present results show that consumption of pistachio as 20% of daily caloric intake leads to significant improvement in HDL and TC/HDL ratio and inhibits LDL cholesterol oxidation. These results suggest that pistachio may be beneficial for both prevention and treatment of coronary artery disease. PMID- 17464103 TI - Drug-induced lymphocyte stimulation test is not useful for the diagnosis of drug induced pneumonia. AB - Diagnosis of drug-induced pneumonia, which represents pulmonary toxicity caused by certain drugs, is difficult, as a large number of different drugs can elicit various immune-mediated diseases with distinct pathomechanisms. The drug-induced lymphocyte stimulation test (DLST) is widely used for diagnosing drug-induced pneumonia in Japan. Recent reports, however, indicate that DLST is not reliable for diagnosis of drug-induced pneumonia. To diagnose drug-induced pneumonia, a provocation test with the suspected drug is the most reliable method of assessing the relationship between the drug and pneumonia. We examined the correlation between the DLST and the provocation test in 6 cases of suspected drug-induced pneumonia. DLST was performed in all of the patients. The causes of pneumonia in all patients were confirmed by a provocation test. The DLST was positive in 3 of 6 cases of suspected drug-induced pneumonia, but the suspected drugs were ruled out by the provocation test. If we had relied solely on the DLST, these 3 cases would have been labeled as false allergy. The results of the DLST did not coincide with the results of the provocation test in any of the cases. Our results suggest that the DLST is not useful for the diagnosis of drug-induced pneumonia. Following provocation with the causative drug, reappearance of pulmonary infiltration was not observed in any of the cases. These findings indicate that a carefully performed provocation test is the safe and most reliable method. PMID- 17464104 TI - Novel anatomical findings of the prostatic gland and the surrounding capsular structures in the normal prostate. AB - With the increase of the patients with prostate cancer, the number of radical prostatectomy increased prominently. Meanwhile, surgeons and pathologists have difficulty regarding appropriate surgical dissection of the prostate and the pathological diagnosis. These problems are derived from uncertainty or misunderstanding about the precise anatomy. In fact, many surgeons are not confident of the structures of the prostatic gland, its surrounding capsules, and the sphincter. Here we investigated the surgical anatomy of the normal prostate to provide beneficial information regarding radical prostatectomy and subsequent pathological diagnosis. A 40 year-old cadaver with a history of sudden cardiac arrest was utilized in this study. Whole pelvic organs were extirpated en bloc and fixed in formalin. Whole mount step sections from the membranous urethra to the seminal vesicle were prepared and histologically examined. It has been reported that the prostatic parenchyma is covered with outside layer (lateral pelvic fascia) and inside layer (prostatic fascia, also known as "capsule"). Here, we show that nearly one third of the anterior surface of the apical region of the prostate (apical prostate) lacks this "capsule". The apical prostate is a mixture of striated muscles, glands, and elastic fibers. Furthermore, the glandular tissue exists within the anterior fibromuscular stroma and some region of the "capsule". Surgeons often try to preserve neurovascular bundles to maintain erectile function; however, other neural tissue was also observed over the entire surface of the prostate. Surgeons must be aware of these complicated anatomical structures when undertaking radical prostatectomy and subsequently diagnosing extra-prostatic extension. PMID- 17464105 TI - Historical trends in human serum levels of perfluorooctanoate and perfluorooctane sulfonate in Shenyang, China. AB - Perfluorooctane sulfonate (PFOS) and perfluorooctanoate (PFOA) are widespread contaminants in the environment, as well as in wildlife and in humans. The PFOS and PFOA concentrations were determined in historical human serum samples collected in Shenyang, China, in 1987 (n=15), 1990 (n=33), 1999 (n=68) and 2002 (n=119). The serum donors were students, faculty members and university workers. Since the serum PFOA and PFOS levels did not follow a normal or log-normal distribution, a nonparametric method was applied to analyze the historical trends. For the total male and female subjects, the median level of serum PFOA increased significantly from 0.08 microg/l in 1987 to 4.3 microg/l in 2002 (p<0.05), while the median level of serum PFOS also increased significantly from 0.03 microg/l in 1987 to 22.4 microg/l in 2002 (p<0.05). Both the serum PFOA and PFOS levels continued to increase from 1999 to 2002, with remarkable increases observed in females: 6.3-fold increase for PFOA and 13-fold increase for PFOS. In 2002, serum PFOA and PFOS concentrations of female subjects have increased to 4.9 microg/l and 22.4 microg/l in median, respectively, which are comparable to those in U.S.A. and Japan. For male subjects, serum PFOA and PFOS concentrations (1.6 microg/l and 8.3 microg/l in median, respectively) are comparable to those in Italy. The data from this study indicate that females are likely to experience higher exposure to these chemicals. PMID- 17464106 TI - Difference in quality of life according to the severity of malocclusion in Japanese orthodontic patients. AB - Although quality of life (QOL) assessment is important in dentistry, it has not been fully investigated in orthodontic patients. We investigated the health related generic QOL (entire body health) and disease specific QOL (oral health) in adult patients with malocclusions at the first visit. One hundred and twenty seven orthodontic patients and 66 persons with normal occlusion were recruited for the study. The subjects were divided into the three following groups based on their treatment: 61 patients in need of surgical correction (SURG), 66 patients in need of non-surgical correction (NONS), and 66 control subjects with normal occlusion. Their dentofacial morphology was assessed using a specific Severity Score (SS), which was set up originally based on their cephalometric radiographs and their plaster models of arrangement of their teeth. The subjects also completed a generic QOL assessment questionnaire, the SF-36, and two disease specific QOL instruments. The patients with malocclusions, especially SURG, had lower disease-specific QOL, although the generic QOL was equal to that of control subjects. Furthermore, in patients with the same severity of dentofacial deformities, especially SS 4 and SS 5, the borderline cases of surgical correction and non-surgical correction, there were differences between SURG and NONS in some items of the QOL. The severity of malocclusion evidently plays an important role in patients' choice of treatment, but also QOL appeared to play a significant role. The QOL assessment may contribute to the selection of the best treatment for improving QOL, especially for borderline cases with moderate degrees of orthodontic abnormality. PMID- 17464107 TI - Development of glomerular endothelial cells, podocytes and mesangial cells in the human fetus and infant. AB - The process of glomerular development consists of four developmental stages: vesicle (V) stage, S-shaped body (S) stage, capillary loop (C) stage and maturation (M) stage. However, the development of glomerular endothelial, mesangial and epithelial cells in fetal and infant kidneys remains unclear. In order to determine the characteristics of human glomerular development, we investigated the process of glomerular development by staining fetal and infant kidneys for CD31, CD34 and FB21, markers for endothelial cells, alpha-smooth muscle actin (alpha-SMA), a marker for mesangial cells, and nephrin, a marker for podocytes. These series of studies were carried out on kidneys obtained at autopsy from 27 fetuses and 5 infants. The fetuses were divided into the following 5 groups according to gestational age; 13-19, 20-24, 25-29, 30-34 and 35-39 weeks. In each group, glomerular development was classified according to the developmental stage and the staining patterns for CD31, CD34, FB21, alpha-SMA and nephrin. The proportion of V-stage development in 100 glomeruli examined was highest at 13-19 weeks. After 20 weeks, the V-stage proportion decreased gradually, and the proportion of S stage became highest at 20-24 weeks. The C stage proportion was highest at 25-29 weeks, while the M-stage proportion was highest in infants aged 1-6 months. The staining patterns for CD31, CD34 and FB21 were similar in endothelial cells after 25 weeks of gestation. Staining of alpha SMA and nephrin was first observed in the S stage. In conclusion, maturation of endothelial cells starts at 25 weeks and is completed by 35 weeks of gestation. Epithelial cells and mesangial cells first appear during the S stage. PMID- 17464109 TI - Pre-renal azotemia: a flawed paradigm in critically ill septic patients? AB - The term pre-renal azotemia (or on occasion 'pre-renal renal failure') is frequently used in textbooks and in the literature to indicate an acute syndrome characterized by the presence of an increase in the blood concentration of nitrogen waste products (urea and creatinine). This syndrome is assumed to be due to loss of glomerular filtration rate but is not considered to be associated with histopathological renal injury. Thus, the term is used to differentiate 'functional' from 'structural' acute kidney injury (AKI) where structural renal injury is taken to indicate the presence of so-called acute tubular necrosis (ATN). This paradigm is well entrenched in nephrology and medicine. However, growing evidence from experimental animal models, systematic analysis of the human and experimental literature shows that this paradigm is not sustained by sufficient evidence when applied to the syndrome of septic AKI, especially in critically ill patients. In such patients, several assumptions associated with the 'pre-renal azotemia paradigm' are violated. In particular, there is no evidence that ATN is the histopathological substrate of septic AKI, there is no evidence that urine tests can discriminate 'functional' from 'structural' AKI, there is no evidence that any proposed differentiation leads or should lead to different treatments, and there is no evidence that relevant experimentation can resolve these uncertainties. Given that septic AKI of critical illness now accounts for close to 50% of cases of severe AKI in developed countries, these observations call into question the validity and usefulness of the 'pre-renal azotemia paradigm' in AKI in general. PMID- 17464110 TI - The concept of acute kidney injury and the RIFLE criteria. AB - Over last half century, the concept of acute renal failure has evolved and with it our estimates of the incidence, prevalence and mortality. Indeed, until very recently no standard definition of acute renal failure was available, and this lack of a common language created confusion and made comparisons all but impossible. In response to the need for a common definition and classification of acute renal failure, the Acute Dialysis Quality Initiative group of experts developed and published a set of consensus criteria for defining and classifying acute renal failure. These criteria which make up acronym 'RIFLE' classify renal dysfunction according to the degree of impairment present: risk (R), injury (I), and failure (F), sustained loss (L) and end-stage kidney disease (E). However, as these criteria were developed, a new concept immerged. Renal dysfunction was no longer only considered significant when it reached the stage of failure, but a spectrum from early risk to long-term failure was recognized and codified. Subsequent studies have validated these criteria in various populations and have shown that relatively mild dysfunction is associated with adverse outcomes. The term acute kidney injury has subsequently been proposed to distinguish this new concept from the older terminology of failure. PMID- 17464111 TI - The liver and the kidney: mutual clearance or mixed intoxication. AB - Hepatorenal syndrome is a frequent complication in patients with cirrhosis, ascites and advanced liver failure. Its annual incidence in patients with ascites has been estimated at 8%. Hepatorenal syndrome is a functional renal failure due to low renal perfusion. Renal histology is normal or shows lesions that do not justify the decrease in glomerular filtration rate. The traditional concept is that hepatorenal syndrome is due to deterioration in circulatory function secondary to an intense vasodilation in the splanchnic circulation (peripheral arterial vasodilation hypothesis). Over the last decade, however, several features have suggested a much more complex pathogenesis. In this article new concepts on the pathogenesis of hepatorenal syndrome are reported, the current options for prophylaxis are shown, and the most applicable treatments are described. PMID- 17464112 TI - Critical care nephrology: a multidisciplinary approach. AB - BACKGROUND/AIMS: Acute renal failure is a common complication in critically ill patients, affecting some 25% of intensive care unit (ICU) admissions, and is associated with high mortality rates of around 40-50%. Acute renal failure in the ICU frequently occurs as part of multiple organ failure (MOF). METHODS: We reviewed the pertinent medical literature related to the occurrence of acute renal failure in the ICU and its association with other organ failures. We also reviewed the literature related to different patient management strategies, notably the differences between 'closed' and 'open' ICU formats. RESULTS: The increasingly common association of acute renal failure with other organ failures, in the context of a more generalized MOF, has important implications on patient care, moving management away from the realm of nephrologists and towards a more multidisciplinary approach. Closed ICU formats with intensivist-led care, supported by specialist consultation, have been shown to be associated with improved ICU outcomes. CONCLUSION: ICU patients with acute renal failure should be managed using a multidisciplinary team approach led by an intensivist. Good collaboration and communication between intensivists and renal and other specialists is essential to insure the best possible care for ICU patients with renal disease. PMID- 17464113 TI - Incidence, classification, and outcomes of acute kidney injury. AB - BACKGROUND: Traditionally the epidemiology of acute renal failure was assessed in patients requiring renal replacement therapy. Recent data emphasized the importance of less severe impairment of kidney function, hence the terminology acute kidney injury (AKI) was introduced. METHODS: In this paper we present a review of current published data on the epidemiology of AKI. RESULTS: The RIFLE classification categorizes the whole severity range of AKI into 3 severity categories and 2 outcome classes. AKI is associated with increased costs and worse outcomes. Increasing severity classes are associated with increasing morbidity and mortality. There is an increasing incidence of AKI, while mortality seems to decrease. CONCLUSION: Small changes in kidney function have an impact on outcomes and this knowledge has led to the introduction of the terminology AKI, encompassing both discrete and severe impairment of kidney function. The RIFLE classification describes the whole range of AKI and has been validated in multiple cohorts. As a consequence of increasing comorbidity, the incidence of AKI is increasing. The incidence of acute renal failure requiring renal replacement therapy even compares to that of acute lung injury, and up to two thirds of general ICU patients meet RIFLE criteria for AKI. PMID- 17464114 TI - Pathophysiology of acute kidney injury: roles of potential inhibitors of inflammation. AB - The pathogenesis of acute kidney injury (AKI) is complex and varies to some extent based on the particular cause. Inflammation contributes to this pathophysiology in a variety of contexts. Inflammation can result in reduction in local blood flow to the outer medulla with adverse consequences on tubule function and viability. Both the innate and adaptive immune responses are important contributors. With ischemia/reperfusion endothelial cells upregulate a number of adhesion molecules which have counterreceptors on leukocytes. A number of vasoactive mediators that are released with injury, such as nitric oxide, may also affect leukocyte- endothelial interactions. Tubule epithelial cells generate proinflammatory and chemotactic cytokines. We and others have found that injection of mesenchymal stem (stromal) cells is protective against renal injury as assessed by serum creatinine measured 24 h after ischemia. The mechanism of such protection may be through intrarenal paracrine effects to decrease inflammation or by systemic immune modulation. Resolvins (Rv) and protectins (PD) have been identified as two newly identified families of naturally occurring n-3 fatty acid docosahexaenoic acid metabolites. In collaboration with Serhan et al. we recently reported that, in response to bilateral ischemia/reperfusion injury, mouse kidneys produce D series resolvins (RvDs) and PD1 [J Immunol 2006;177:5902 5911]. Administration of RvDs or PD1 to mice prior to, or subsequent to, ischemia resulted in a reduction in functional and morphological kidney injury. Understanding how these anti-inflammatory processes are regulated may provide insight into how we might intervene to facilitate and enhance them so that we might prevent or mitigate the devastating consequences of AKI. PMID- 17464115 TI - Sepsis and multiple organ failure. AB - BACKGROUND/AIMS: Sepsis and multiple organ failure are complex processes that result from dysregulation of the immune response and its associated hematological, hemodynamic and metabolic disturbances. METHODS: Review of the pathophysiological basis for sepsis and a review of the literature on its mechanisms of expression. RESULTS: Sepsis is the host response to an injury, often infectious in origin, that creates both pro- and anti-inflammatory immune responses. The level and duration of this response roughly correlates with outcome. Subcellular injury characterized by increased oxidative stress defines the central mitochondrial component of this process. Treatments which minimize the amplification of this response are usually more effective at reducing tissue injury than are measures aimed at suppressing the inflammatory response. CONCLUSIONS: Sepsis is a complex process whose expression and treatment are just now being defined. Treatments that minimize the overall host response still represent the most effective strategies. PMID- 17464116 TI - Classification, incidence, and outcomes of sepsis and multiple organ failure. AB - BACKGROUND/AIMS: Sepsis and multiple organ failure are common complications in intensive care unit (ICU) patients and are associated with considerable morbidity and mortality. METHODS: We reviewed pertinent medical literature related to sepsis and multiple organ failure to determine strategies of classification, the current incidence, and the outcomes associated with these disease processes. RESULTS: Sepsis affects some 40% of ICU admissions, severe sepsis occurs in about 30%, and septic shock in 15%. Recent consensus has improved the definition of sepsis and proposed a new classification system based on predisposing factors, infection, immune response, and organ dysfunction. We discuss the possible components of each of these four categories. CONCLUSION: Although there is some evidence that mortality rates may have decreased in recent years, the incidence of sepsis is increasing so that overall deaths from this disease are increasing. Improved diagnostic techniques and classification may help target therapies more rapidly and more appropriately. PMID- 17464117 TI - Genetic polymorphisms in sepsis- and cardiopulmonary bypass-associated acute kidney injury. AB - Acute kidney injury (AKI) is a major medical problem in critical illness, and has a separate independent effect on the risk of death. Septic shock and cardiac surgery utilizing cardiopulmonary bypass are the two most common factors contributing to AKI. Clinical predictors and biochemical markers identified for the development of AKI can only explain a part of this individual risk. Another tool to predict the risk of AKI and to improve individualized patient care focuses on the identification of genetic risk factors which might be involved in the development of AKI. However, to date our knowledge on the importance of such genetic polymorphisms in influencing the susceptibility to and severity of AKI remains limited. There is evidence that several genetic polymorphisms accounting for sepsis- or cardiopulmonary bypass-associated AKI involve genes which participate in the control of inflammatory or vasomotor processes. In this article, we will review current knowledge concerning the role of genetic polymorphism in the pathogenesis of sepsis- and cardiopulmonary bypass-associated AKI and discuss possible areas for future developments and research in this field. PMID- 17464118 TI - Predictive capacity of severity scoring systems in the ICU. AB - Severity scoring systems were first introduced to intensive care units (ICUs) in 1980. The basis for their development was the intention to provide information on the prognosis of patients, the efficacy of therapeutic interventions, stratification for clinical studies, workload and benchmarking of ICUs. Despite the appearance of several specialized scoring systems, the general mortality prediction systems such as APACHE, SAPS and MPM scores and their constantly improved successors have become the most popular and widely tested models. The newest development in this field is SAPS III which is the first 'global' model using a data set acquired from 307 ICUs from all over the world. PMID- 17464119 TI - Determining the degree of immunodysregulation in sepsis. AB - During sepsis, the anti-infectious response is closely linked to an overwhelming inflammatory process. The latter is illustrated by the presence in plasma of numerous inflammatory cytokines, markers of cellular stress (e.g. high mobility group box-1 protein), complement-derived compounds (e.g. anaphylatoxin C5a), lipid mediators, and activated coagulation factors. All mediators contribute in synergy to tissue injury, organ dysfunction, and possibly to lethality. To dampen this overzealous process, a counter-regulatory loop is initiated. The anti inflammatory counterpart involves few anti-inflammatory cytokines (e.g. interleukin-10, transforming growth factor-beta), numerous neuromediators (e.g. adrenalin, acetylcholine), and some other factors (e.g. heat shock proteins, ligand of TREM-2, adenosine). These mediators modify the immune status of circulating leukocytes as illustrated by their decreased cell-surface expression of HLA-DR or their reduced ex vivo pro-inflammatory cytokine production in response to Toll-like receptor agonists (e.g. endotoxin, lipoproteins). However, circulating leukocytes remain responsive to whole bacteria and produce normal or even enhanced levels of anti-inflammatory cytokines. Thus, the immune dysregulation observed in sepsis corresponds to a reprogramming of circulating leukocytes. PMID- 17464120 TI - Nutritional management in acute illness and acute kidney insufficiency. AB - There are now powerful compensatory therapies to counteract kidney deficiency and the prognosis of patients with acute renal failure is mainly related to the severity of the initial disease. Renal failure is accompanied by an increase in both severity and duration of the catabolic phase leading to stronger catabolic consequences. The specificity of the metabolic and nutritional disorders in the most severely ill patients is the consequence of three additive phenomena: (1) the metabolic response to stress and to organ dysfunction, (2) the lack of normal kidney function and (3) the interference with the renal treatment (hemodialysis, hemofiltration or both, continuous or intermittent, lactate or bicarbonate buffer, etc.). As in many other diseases of similar severity, adequate nutritional support in acutely ill patients with ARF is of great interest in clinical practice, although the real improvement as a result of this support is still difficult to assess in terms of morbidity or mortality. PMID- 17464121 TI - Fundamentals of oxygen delivery. AB - Oxygen is vital to life. A series of steps are needed to transport oxygen from the lungs to the mitochondrion where the bulk of it is used for generation of energy. Understanding this pathway, which still remains to be properly characterized, will greatly aid both diagnosis and management of the hypoxic patient. PMID- 17464122 TI - Principals of hemodynamic monitoring. AB - BACKGROUND/AIMS: Hemodynamic monitoring is the cornerstone of patient management in the intensive care unit. However, to be used effectively its applications and limitations need to be defined and its values applied within the context of proven therapeutic approaches. METHODS: Review of the physiological basis for monitoring and a review of the literature on its utility in altering patient outcomes. RESULTS: Most forms of monitoring are used to prevent cardiovascular deterioration or restore cardiovascular wellness. However, little data support the generalized use of aggressive resuscitation protocols in all but the most acutely ill prior to the onset of organ injury. Outcomes improve with aggressive resuscitation in some patients presenting with early severe sepsis and in postoperative high-risk surgical patients. CONCLUSIONS: Monitoring should be targeted to meet the specific needs of the patient and should not be applied in a broad fashion and whenever possible it should be used as part of a treatment protocol of proven efficacy. PMID- 17464123 TI - Acid-base disorders and strong ion gap. AB - The application of modern quantitative physical chemical techniques to clinical acid-base has yielded important new information about the nature and clinical significance of metabolic acid-base disorders. Abnormalities identified by the strong ion gap appear to be common in critically ill patients and are associated with increased mortality especially when identified early in the course of critical illness. Attempts to identify the exact chemical nature of ions identified by the strong ion gap have only been of limited success and further study is needed. PMID- 17464124 TI - Fluid resuscitation and the septic kidney: the evidence. AB - Acute kidney injury (AKI) is a common complication of severe sepsis. Severe sepsis is the most common cause of AKI in ICU. The widely accepted and practiced initial cornerstone of treatment for septic AKI is fluid resuscitation. The biological rationale for fluid resuscitation in septic AKI is based on the assumption that septic AKI is an ischemic form of AKI and that increasing renal perfusion and oxygen delivery by means of fluid resuscitation will protect the kidney. Whether this is true, however, remains uncertain. In this paper, we discuss salient pathophysiological aspects of AKI, review the evidence available on the need for fluid resuscitation, the amount and the type of fluid that might be best suited to AKI and discuss all major aspects of fluid resuscitation for septic AKI in humans and experimental animals. PMID- 17464125 TI - Factors affecting circuit patency and filter 'life'. AB - Frequent clotting applying continuous renal replacement therapy means treatment can be inadequate and with increased costs for circuits and nursing time. Patency of the extracorporeal circuit is commonly achieved using anticoagulants such as heparin. When anticoagulants are not used, or clotting occurs within a few hours of use, with anticoagulation, blood flow failure is a likely cause. The blood pump can fail to deliver without operator awareness. Clotting within the membrane and/or venous 'air-trap' chamber is common where resistance to blood flow is high with stasis and turbulence. The design of the venous chamber allows the blood fill level to oscillate and form a clot, with a blood filter at the exit of the chamber also causing clot development. Several practices attempt to prevent clotting, however most without evidence. Adding heparin to the circuit during the preparation phase, ensuring that the access catheter is not obstructed, a blood flow setting of >or=200 ml/min, and administration of substitution fluids before the membrane (predilution) can be useful strategies for increasing circuit patency. An audit of filter life is useful and necessary feedback to nursing staff training strategies. This promotes safety and, when circuit patency is poor, may reflect poor troubleshooting ability. PMID- 17464126 TI - Starting up a continuous renal replacement therapy program on ICU. AB - BACKGROUND/AIM: The questions as to which treatment is the most effective for the replacement of renal function in critically ill patients with acute renal failure and the qualifications needed by nurses to manage the continuous renal replacement therapy (CRRT) device are part of an ongoing debate between nephrologists and intensivists, between nurses of the renal ward and the ICU. METHODS: The keys to a successful CRRT program are a well-balanced and practical education program, a user-friendly dialysis machine, and technical support 24 h/day. A computerized data management system will diminish the workload to an acceptable level. RESULTS: Intensive care nurses on our ICUs are well trained to execute CRRT without the involvement of nephrology nurses. On the ICU, the 24 hour presence of an intensivist is an additional advantage to solve medical problems involving CRRT. The daily cost of CRRT is only dependent on the devices and independent of human resources. CONCLUSION: Initiating and maintaining a CRRT program is a great challenge for the ICU nurse. The possible problems remain within the ICU staff's ability to solve if they follow an education program. If the workload for the nurses is well monitored, extra personnel can be avoided. PMID- 17464127 TI - Is there a need for a nurse emergency team for continuous renal replacement therapy? AB - The use of an emergency response team for unwell patients has provided an improvement in hospital care standards by reducing medical and postoperative adverse outcomes. Use of a nurse emergency team for patients treated with continuous renal replacement therapy (CRRT) also has potential to reduce adverse outcomes with CRRT, where staff may lack experience or find troubleshooting CRRT difficult in an ICU with many critically ill patients in their care. Differing nursing models are used to provide CRRT in the ICU, and all of these could benefit from a nursing response team at some time. The response must be immediate, with suitably available and CRRT-experienced nurses. As with medical emergency team use, the nursing emergency team for CRRT would be called when a deviation from a standard criterion list occurs. The list could include: prolonged blood pump stoppage (approximately 2 min); air detection alarm; blood leakage; sudden circuit pressure changes-transmembrane pressure (>200 mm Hg) or venous pressure (>200 mm Hg) or arterial pressure negative (>or=100 mm Hg); the need to override a fluid balance alarm 3 times in 5 min; patient hypotension; cardiac arrest or similar event, or the nurse is concerned that the machine is malfunctioning. The 'human resource' is the biggest challenge to developing a suitable response team 24/7, however where ICU and nephrology nurses work in a collaborative approach for CRRT, a response team would be more easily established and may not be required continuously. PMID- 17464128 TI - Information technology for CRRT and dose delivery calculator. AB - BACKGROUND: The application of information technology (IT) to the field of critical care nephrology is a process that may reduce errors in care delivery, improve monitoring, decrease unintentional practice variation, increase the quality and accuracy of delivered treatments. METHODS: This review presents some examples of potential applications of recent IT achievements to clinical practice. RESULTS: The adequacy calculator for continuous therapy dose prescription was recently shown to accurately predict urea clearance. When clearances above 60 ml/min where prescribed, the calculator tended to overestimate effective clearances; this overestimation generally remained within an error of 15%. Nevertheless, the delivered Kt/V in 24 h will always approach the target value of 1.2. The use of the calculator enabled strict monitoring of treatments. Furthermore, the so-called 'next generation' machines have technical characteristics in common that allow the highest safety and accuracy levels: some of these aspects are addressed and commented on in the present review. CONCLUSION: IT is having and will likely have a significant impact on patient safety, practice variation, patient assessment and monitoring, and documentation of the demographics of acute renal failure and dialysis. One of the most recent and potentially interesting aspects of IT implementation on acute dialysis might be the renal replacement dose monitoring and calculation: close control of the therapy delivery and, eventually, prescription adjustments might be optimized. PMID- 17464129 TI - Emerging biomarkers of acute kidney injury. AB - BACKGROUND: Acute kidney injury (AKI) is a major clinical problem with a rising incidence and high mortality rate. The lack of early biomarkers has resulted in an unacceptable delay in initiating therapies. METHODS: Here we will update the reader on promising new blood and urinary biomarkers that have recently emerged through the application of innovative technologies such as functional genomics and proteomics to human and animal models of AKI. RESULTS: The most promising biomarkers of AKI for clinical use include a plasma panel (NGAL and cystatin C) and a urine panel (NGAL, Il-18 and KIM-1). CONCLUSIONS: As they represent tandem biomarkers, it is likely that the AKI panels will be useful for timing the initial insult and assessing the duration and severity of AKI. Based on the differential expression of the biomarkers, it is also likely that the AKI panels will distinguish between the various types and etiologies of AKI. It will be important in future studies to validate the sensitivity and specificity of these biomarker panels in clinical samples from large cohorts and from multiple clinical situations. PMID- 17464130 TI - Diagnosis of acute kidney injury: from classic parameters to new biomarkers. AB - A change in serum creatinine is the standard metric used to define and monitor the progression of acute kidney injury (AKI). This marker is inadequate for a number of reasons including the fact that changes in serum creatinine are delayed in time after kidney injury and hence creatinine is not a good indicator to use in order to target therapy in a timely fashion. There is an urgent need for early biomarkers for the diagnosis of AKI. There is also a need for biomarkers that will be predictive of outcome and which can be used to monitor therapy. There are a limited number of biomarkers that are being validated by a number of groups and from this list clinically useful reagents are likely to be derived over the next few years. In this article the status of 5 potential urinary biomarkers for AKI are discussed: kidney injury molecule-1, N-acetyl-Beta-D-glucosaminidase, neutrophil gelatinase-associated lipocalin, cystatin C, and interleukin-18. Considerable progress has been made although much continues to be needed to validate these markers for routine clinical use. Armed with these new tools the future will look much brighter for the patient with AKI as it is likely that early diagnosis and better predictors of outcome will lead to new therapies which can be introduced earlier in the course of disease. PMID- 17464131 TI - Endotoxin and cytokine detection systems as biomarkers for sepsis-induced renal injury. AB - BACKGROUND: A reliable biomarker as an indicator of the presence of severe sepsis is an unmet medical need. METHODS: Review of recent literature on this topic focusing upon endotoxin and cytokine assays. RESULTS: The ideal biomarker for sepsis would be readily available, technically easy to perform with a quick turn around time, inexpensive, highly specific, very sensitive, and preferably highly correlated in quantitative terms with disease severity. Such a test would provide early diagnostic accuracy, prognostic information, and indicate responsiveness to treatment interventions. Regrettably no such biomarker exists for sepsis at present, and it is not likely that such an ideal assay will be developed in the foreseeable future. CONCLUSIONS: Despite their shortcomings, a number of existing and candidate biomarker assays are available and can provide some useful information to the clinician caring for septic patients. The relative merits of endotoxin measurement, interleukin-6 levels and a variety of other sepsis markers are reviewed. Full implementation of these biomarkers may improve diagnostic accuracy over the standard clinical criteria for sepsis. PMID- 17464132 TI - Quantifying dynamic kidney processes utilizing multi-photon microscopy. AB - Multi-photon microscopy and advances in optics, computer sciences, and the available labeling fluorophores now allow investigators to study the dynamic events within the functioning kidney with subcellular resolution. This emerging technology, with improved spatial and temporal resolution and sensitivity, enables investigators to follow complex heterogenous processes in organs such as the kidney. Repeated determinations within the same animal are possible minimizing their use and inter-animal variability. Furthermore, the ability to obtain volumetric data (3D) makes quantitative 4D (time) analysis possible. Finally, use of up to three fluorophores concurrently allows three different or interactive processes to be observed simultaneously. Therefore, this approach compliments existing molecular, biochemical, and pharmacologic techniques by advancing data analysis and interpretation to subcellular levels for molecules without the requirement for fixation. PMID- 17464133 TI - Diuretics in the management of acute kidney injury: a multinational survey. AB - BACKGROUND: Diuretics are a common intervention in critically ill patients with acute kidney injury (AKI). However, there is no information that describes the practice patterns of diuretic use by clinicians. METHODS: Multinational, multicenter survey of intensive care and nephrology clinicians that utilized an 18-question self-reported questionnaire. RESULTS: The survey generated 331 responses from 16 countries. Academic institutions comprised 77.5%, with the remaining being from metropolitan, regional or private hospitals. The use of furosemide was most common (67.1%), delivered primarily intravenously (71.9%) and by bolus dosing (43.3%). Other diuretics were infrequently used. The majority rated current serum creatinine (73.6%) and urine output (73.4%), blood pressure (59.7%), central venous pressure (65.2%) and risk of toxicity (62.4%) important when deciding on a dose. Pulmonary edema was a prime physiologic indication for diuretic use (86.3%). Diuretic use was also common with rhabdomyolysis (55.6%), major surgery (56%), and cardiogenic shock (56.2%), and sepsis (49.5%). Diuretic use was most commonly given either prior to (57.7%) or during recovery (33.9%) after renal replacement therapy (RRT). Most (76.6%) targeted a diuresis of >or=0.5-1.0 ml/kg/h. The majority did not believe that diuretics could reduce mortality (74.3%), reduce need for (50.8%) or duration of RRT (57.8%) or improve renal recovery (68.2%), however, many stated uncertainty. Most (85.1%) would be willing to participate in a randomized trial (RCT) of diuretics in AKI with 72.4% believing it ethically acceptable to allocate patients to placebo. CONCLUSION: Diuretics are frequently used in AKI. Clinicians are most familiar with furosemide given intravenously and titrated to a physiologic endpoint of urine output. Most clinicians believe an RCT on diuretic use in AKI is justified and ethical. This survey confirms clinical agreement and a need for higher quality evidence on diuretic use in AKI. PMID- 17464134 TI - Stem cells in acute kidney injury. AB - The susceptibility of developing acute renal failure depends on the ability of the kidney to recover from acute injury and regain normal function. Recently, the possible contribution of stem cells (SCs) to the regeneration of acute tubular injury has been investigated. There is evidence indicating that, under pathophysiological conditions, SCs derived from bone marrow are able to migrate in the injured kidney but they seem to play a minor role in tubular regeneration in regard to the resident cells. However, the administration of ex vivo expanded bone marrow-derived mesenchymal SCs has proven to be beneficial in various experimental models of acute renal failure. The mechanism underlining this beneficial effect is still matter of debate. The transdifferentiation or fusion of SCs to repopulate tubules is considered to play a minor role. The administered SCs may, however, modify the microenvironment by inducing dedifferentiation and proliferation of tubular cells surviving to injury or by allowing expansion of resident SCs. The recent identification of resident progenitor/SC populations in the adult kidney supports the hypothesis that resident SCs may play a critical role in the repair of renal injury. Therefore, therapeutic strategies to exploit the regenerative potential of SCs may be based on the administration of ex vivo expanded SCs or on stimulation of expansion and differentiation of local progenitor/SC populations. PMID- 17464135 TI - Anticoagulation options for patients with heparin-induced thrombocytopenia requiring renal support in the intensive care unit. AB - World wide, heparins are the most commonly used anticoagulants for renal replacement therapy (RRT). In the intensive care unit (ICU) keeping the RRT circuit patent is more difficult than during routine outpatient hemodialysis, as ICU patients typically have sepsis and/or inflammation resulting in activation of the procoagulant pathways, with reduced antithrombin. One important cause of repeated RRT circuit clotting is heparin-induced thrombocytopenia (HIT), which should not be overlooked in patients with a reduced platelet count. If HIT is clinically suspected then all heparins should be withdrawn, and the patient systemically anticoagulated with either a direct thrombin inhibitor, such as argatroban and/or hirudin, or the heparinoid danaparoid. The availability and licensing of these alternative anticoagulants varies from country to country. Argatroban has to be continuously infused, which is an advantage for continuous RRT, but not for intermittent RRT, and can be monitored by activated partial thromboplastin time. Hirudin has a prolonged half life, which is extended by hirudin antibodies, and requires specialist monitoring to prevent over anticoagulation. Although the half life of danaparoid is increased in renal failure, it can be given as boluses for intermittent and continuous RRT, or by continuous infusion during continuous RRT, but requires factor Xa monitoring. PMID- 17464136 TI - Nutritional support during renal replacement therapy. AB - BACKGROUND/AIMS: Malnutrition is common in critically ill patients with acute renal failure. The aim of this review is to describe the basis for nutritional support during renal replacement therapy. METHODS: Review of the literature. RESULTS: Techniques of nutritional support and nutritional requirements are described. CONCLUSION: Early aggressive enteral, parenteral or combine nutritional support is required in critically ill patients on replacement therapy. PMID- 17464137 TI - Vascular access for HD and CRRT. AB - A good functioning vascular access is an essential component for adequate renal replacement therapy (RRT) in acute kidney injury. Tunneled, cuffed catheters are preferred if the anticipated duration of RRT is more than 3 weeks. The right jugular vein is the preferred insertion site for the temporary dialysis catheter (TDC), with ultrasound-guided insertion reducing the risk of mechanical complications. The femoral vein is the second choice, whereas the subclavian vein should be avoided. The most important complications of a TDC are acute malfunction and infection. Intraluminal thrombosis, fibrin sleeve formation, malpositioning and kinking result in acute malfunction. Recirculation can be reduced by correct placement of the catheter and is more an issue for intermittent hemodialysis than for continuous RRT. Strict adherence to simple preventive strategies reduces catheter-related bloodstream infection. In selected patients more sophisticated strategies such as the use of antibiotic/antiseptic impregnated catheters and antibiotic/antiseptic lock solutions may be useful. PMID- 17464138 TI - Dialysate and replacement fluid composition for CRRT. AB - Continuous renal replacement therapies (CRRTs) are increasingly used in order to maintain normal or near-normal acid-base balance in intensive care unit (ICU) patients. Acid-base balance is greatly influenced by the type of dialysis employed and by the administration route of replacement fluids. In continuous veno-venous hemofiltration, buffer balance depends on losses with ultrafiltrate and gain with replacement fluid, while in techniques such as continuous veno venous hemodiafiltration, clinicians should balance the role of the dialysate. The type of buffer greatly influences not only acid-base correction, but also clinical outcome. Lactate or bicarbonate fluids are currently used, but recent studies suggest that bicarbonate-buffered replacement fluids can improve acid base status and reduce cardiovascular events better than lactate fluids. The buffer concentration should exert a buffer load that may compensate for deficits, for losses in the buffer process, and for extracorporeal losses and should therefore usually be supraphysiological. However, the dialysate buffer or electrolyte concentration need always to be balanced with that of the replacement fluids employed. Both fluids should contain electrolytes in concentrations aiming for a physiologic level and taking into account preexisting deficits or excess and all input and losses. Clinicians should be aware that in CRRTs the quality control for sterility, physical properties, individualized prescription and balance control are vitally important. PMID- 17464139 TI - Results from international questionnaires. AB - BACKGROUND: The practice of renal replacement therapy (RRT) has reached an optimal standard of care worldwide. Nevertheless, some aspects of acute renal failure treatment and support still present wide variability between different centers. This is especially true for the mode and dose of RRT. This review describes the epidemiology of dialysis prescription and delivery around the world based on recent observational studies and international surveys. RESULTS: Continuous RRT is delivered in 80% of intensive care units around the world. Since a certain consensus has been achieved on the adequacy of 35 ml/kg/h of clearance in continuous therapies, recent observations based on questionnaires and surveys demonstrated that such adequate therapy was only prescribed in the minority of patients. The number of centers prescribing adequate dialysis dose is increasing, but there are still many institutions where prescription is made with no specific adequacy targets and effective delivery is not measured. Several barriers to reaching adequacy targets have been identified including the lack of a high evidence multicentric trial, logistics, costs, personnel and technical difficulties. CONCLUSION: A trend to continuous therapies and increased RRT dosage over the last 10 years is shown by the surveys presented, even if scientific evidence is now very necessary as far as definitive RRT indications and prescriptions are concerned. PMID- 17464140 TI - Intermittent hemodialysis for renal replacement therapy in intensive care: new evidence for old truths. AB - Acute renal failure requiring dialysis is a frequent complication in critically ill patients with a high morbidity and mortality. Until recently, no evidence based guidelines on the optimal treatment modality for renal replacement in the ICU could be issued because of a lack of well-performed randomized controlled trials (RCT). Over the last years however, some important new concepts and RCTs have been published on this topic. An important concept is the understanding that 'chronic dialysis strategies' are not suitable for acute renal failure patients in the ICU. From this understanding the necessity of daily dialysis followed, and later on, the need for flexible treatments related to the patients' need, using slow long extended daily dialysis (SLEDD). Several recent papers compared continuous renal replacement therapy and intermittent hemodialysis (IHD) in ICU patients, pointing to a lack of differences in outcome, but there were less practical problems using IHD, even in unstable patients. In conclusion, it can be stated that all patients can be treated with IHD when available, without jeopardizing their outcome. Slow extended daily dialysis emerged as a hybrid renal replacement therapeutic modality and has promising features because it combines the advantages of both continuous renal replacement therapy and IHD, but until now, no studies evaluating whether SLEDD is superior to 'regular IHD' are available. PMID- 17464141 TI - Continuous renal replacement in critical illness. AB - Acute renal failure in the intensive care unit is usually part of the multiple organ dysfunction syndrome, and the complexity of illness in patients with this complication has risen in recent years. Continuous renal replacement therapy (CRRT) was introduced in the late 1970s and early 1980s to compensate for the inadequacies of conventional intermittent hemodialysis (IHD) in the treatment of these patients. IHD was considered aggressive and unphysiological, often resulting in hemodynamic intolerance and limited efficiency. Although CRRT has been shown to be physiologically superior with respect to IHD in both observational and randomized studies, it is not clear whether this physiological superiority translates into clinically important gains. A number of recent studies have tried to address this issue, and with these, there is a lack of evidence to suggest improved survival and major clinical outcomes with CRRT. However, these studies are generally underpowered and have certain aspects which may influence the interpretation of their results. In addition, the development of hybrid techniques, such as slow extended daily dialysis, makes this a dynamic area of study where the terms of comparison are constantly changing. This article reviews recent trials comparing CRRT and IHD, and discusses their results and limitations. PMID- 17464142 TI - Sustained low-efficiency dialysis. AB - Sustained low-efficiency dialysis (SLED) is an increasingly popular form of renal replacement therapy for patients with renal failure in the intensive care unit. Advantages of SLED are efficient clearance of small solutes, good hemodynamic tolerability, flexible treatment schedules, and reduced costs. Studies comparing outcomes of SLED with those of other dialysis modalities are being performed. PMID- 17464143 TI - The role of the International Society of Nephrology/Renal Disaster Relief Task Force in the rescue of renal disaster victims. AB - Disasters are a major cause of distress and material as well as corporal damage. Next to direct trauma, the crush syndrome inducing multiorgan problems as a consequence of muscle compression and the release of muscular contents into the bloodstream is the most important cause of death; this is to a large extent related to the induction of severe acute kidney injury, for which dialysis is a life-saving therapy. The practical means (both hardware and personnel) to do so are, however, often lacking in disaster conditions. The Renal Disaster Relief Task Force (RDRTF) offered support for renal problems in the aftermath of several disasters, e.g. the Marmara earthquake (1999) in Turkey, the Bam earthquake (2003) in Iran, and the Kashmir earthquake (2005) in Pakistan. A preconceived intervention plan is followed with adaptations according to local conditions. Material and personnel are dispatched to the disaster areas. These interventions have been life-saving for a substantial number of victims. The current article describes the structure and approach of the RDRTF. PMID- 17464144 TI - Renal replacement therapy for the patient with acute traumatic brain injury and severe acute kidney injury. AB - Fortunately with improvements in initial medical resuscitation, such as the avoidance of nephrotoxins, the incidence of acute kidney injury requiring renal support in patients with acute traumatic brain injury remains low. However the incidence of cerebral hemorrhage in patients on chronic dialysis programs appears to be increasing. By carefully adapting renal replacement to minimize cardiovascular instability and reduce the rate of change of serum osmolality, patient survival in this group of critically ill patients is increasing and starting to approach that of patients with traumatic brain injury without kidney injury. PMID- 17464145 TI - Cardiopulmonary bypass-associated acute kidney injury: a pigment nephropathy? AB - Acute kidney injury (AKI) is a common and serious postoperative complication following exposure to cardiopulmonary bypass (CPB). Several mechanisms have been proposed by which the kidney can be damaged and interventional studies addressing known targets of renal injury have been undertaken in an attempt to prevent or attenuate CPB-associated AKI. However, no definitive strategy appears to protect a broad heterogeneous population of cardiac surgery patients from CPB-associated AKI. Although the association between hemoglobinuria and the development of AKI was recognized many years ago, this idea has not been sufficiently acknowledged in past and current clinical research in the context of cardiac surgery-related AKI. Hemoglobin-induced renal injury may be a major contributor to CPB-associated AKI. Accordingly, we now describe in detail the mechanisms by which hemoglobinuria may induce renal injury and raise the question as to whether CPB associated AKI may actually be, in a significant part, a form of pigment nephropathy where hemoglobin is the pigment responsible for renal injury. If CPB associated AKI is a pigment nephropathy, alkalinization of urine with sodium bicarbonate might protect from: (1) tubular cast formation from met-hemoglobin; (2) proximal tubular cell necrosis by reduced endocytotic hemoglobin uptake, and (3) free iron-mediated radical oxygen species production and related injury. Sodium bicarbonate is safe, simple to administer and inexpensive. If part of AKI after CPB is truly secondary to hemoglobin-induced pigment nephropathy, prophylactic sodium bicarbonate infusion might help attenuate it. A trial of such treatment might be a reasonable future investigation in higher risk patients receiving CPB. PMID- 17464146 TI - CRRT technology and logistics: is there a role for a medical emergency team in CRRT? AB - Implementing continuous renal replacement therapy (CRRT) in a intensive care unit (ICU) is a somewhat difficult issue and quiet different from starting a new ventilation mode or a new hemodynamic device. It may indeed require an on-call medical emergency CRRT team as expertise in this field is really a key issue to success. Education for the nursing team is another key point, especially as ongoing or continuous education is changing very quickly. Uniformity of the type of device used is another crucial part in the organization process with regard to CRRT implementation in the ICU. Involvement of both the ICU and nephrology teams is another key to success especially when different modes and higher exchange rates are used. Also, a nursing group devoted to the ongoing implementation and education of the ICU team is very useful in order to attain the goals that have been set. Already in 1984 acute renal failure was described as one of the remaining and challenging problems in the ICU. Hemodialysis was not always feasible then because of the hemodynamic instability of critically ill patients. Under those circumstances continuous arteriovenous hemofiltration (CAVH) was advocated as an efficient alternative method with less detrimental hemodynamic effects. At the time it was thought that CAVH would be found to be an effective 'artificial kidney' (control of body fluid, electrolyte and acid-base homeostasis and uremia) and this without serious side effects. But already nearly 25 years ago, it was found that continuous anticoagulation was a major problem that could cause life-threatening complications in posttraumatic and surgical patients. At the time, it was thought that running a protamine infusion on the venous line would help to diminish these complications. CRRT has been carried out in our ICU since 1985, first with CAVH and since 1989 with some early forms of continuous veno-venous hemofiltration (CVVH). The unit has used BSM 22, BM 25 and Prisma for nearly 10 years, and Aquarius since the end of 2001. The educational process started at the beginning of 1990 with the implementation of CVVH using BSM 22 and BM 25. Very soon it was realized that a new strategy implementing pulse high volume hemofiltration (pulse-HVHF) was really needed. Therefore, a nursing group composed of 5-8 nurses who would be taught beforehand was started, and this dedicated group would then teach the rest CRRT Technology and Logistics 355 of the staff nurses. This group exists today and has at least 6-8 meetings/year in which all the problems that must be faced in the implementation of CRRT are dealt with. Here all the steps made by our and other units in this field will be discussed, including an overview of the various protocols implemented and a description of our dedicated nursing group with regard to CRRT. PMID- 17464147 TI - Continuous hemodiafiltration with cytokine-adsorbing hemofilter in the treatment of severe sepsis and septic shock. AB - Continuous hemodiafiltration (CHDF) using a polymethymethacrylate (PMMA) membrane hemofilter (PMMA-CHDF) can effectively and continuously remove various cytokines from the circulating blood. PMMA-CHDF can decrease the blood levels of various cytokines when the blood levels of cytokines are high prior to the initiation of CHDF. The main mechanism of cytokine removal with PMMA-CHDF is the adsorption of cytokines to the hemofilter membrane and this characteristic was not observed in the other membrane material. PMMA-CHDF could improve blood pressure, the depressed monocytic HLA-DR expression, and recover the delayed neutrophil apoptosis in septic patients. Thus, cytokine removal with PMMA-CHDF would be effective for the treatment of severe sepsis and septic shock. PMID- 17464148 TI - Blood and plasma treatments: high-volume hemofiltration--a global view. AB - From the recent past, hemofiltration, particularly high-volume hemofiltration, has rapidly evolved from a somewhat experimental treatment to a potentially effective 'adjunctive' therapy in severe septic shock and especially refractory or catecholamine-resistant hypodynamic septic shock. Nonetheless, this approach lacks prospective randomized studies (PRTs) evaluating the critical role of early hemofiltration in sepsis. An important milestone, which could be called the 'big bang' in terms of hemofiltration, was the publication of a PRT in patients with acute renal failure (ARF). Before this study, nobody believed that hemofiltration could change the survival rate in intensive care. Since that big bang, many physicians consider that hemofiltration at a certain dose can change the survival rate in intensive care. We now must try to define what the exact dose in septic ARF should be. As suggested by many studies this dose might well be higher than 35 ml/kg/h in the septic ARF group. The issue of the dosage of continuous high volume hemofiltration must be tested in future randomized studies. Since the Vicenza study has shown that 35 ml/kg/h is the best dose in terms of survival when dealing with nonseptic ARF in the intensive care unit (ICU), several studies from different groups have shown that a higher dose might be correlated with better survival in septic ARF. This has also been shown in some way by the Vicenza group but not with a statistically significant value. New PRTs have just started in Europe such as the IVOIRE (hIgh VOlume in Intensive Care) study. The RENAL study is another large study looking more basically at dose in nonseptic ARF in Australasia. The ATN study in the USA is also testing the importance of dose in the treatment for ARF. Nevertheless, 'early goal-directed hemofiltration therapy' has to be studied in our critically ill patients. Regarding this issue, fewer studies, mainly retrospective, exist; but again the IVOIRE study will address this issue by studying septic patients with acute renal injury according to the RIFLE classification. This chapter will focus on the early application and adequate dose of continuous high-volume hemofiltration in septic shock in order to improve not only the hemodynamics but also survival in this very severely ill cohort of patients. This could be called the big bang of hemofiltration as one could have never anticipated that an adequate dose of hemofiltration could markedly influence the survival rate of septic ARF patients in the ICU. Apart from the use of an early and adequate dose of Honore/Joannes-Boyau/Gressens 372 hemofiltration in sepsis, a higher dose could also provide a better renal recovery rate and reduce the risk of associate chronic dialysis in these patients. Furthermore, this presentation will also review brand-new papers regarding the use of hemofiltration in systemic inflammatory response syndrome and out-of-hospital cardiac arrest. PMID- 17464149 TI - Blood and plasma treatments: the rationale of high-volume hemofiltration. AB - Since the early 1990s, experts in the field have thought that a reduction in cytokines in the blood compartment could, in theory, reduce mortality, but this is perhaps too naive as the pharmacodynamics and pharmacokinetics of cytokines throughout the body are not well known and are probably much more complicated than previously thought. This ha now led to three leading theories and concepts. Ronco and Bellomo conceived the peak concentration hypothesis in which clinicians concentrate their efforts to remove mediators and cytokines from the blood compartment at the proinflammatory phase of sepsis. By reducing the amount of free cytokines, it is hoped that the level of remote organ (associated) damages can be dramatically decreased and, as a consequence, the overall death rate. In this regard, it is still not known what will happen at the interstitial and tissue level with regard to mediators and cytokines which are obviously the most important part in terms of consequences at the tissue level. In this setting, techniques that can more rapidly and substantially remove great amounts of cytokines or mediators are privileged. Among these, there is high-volume and very high-volume hemofiltration and a number of hybrid therapies encompassing high permeability hemofiltration, super high-flux hemofiltration, hemo-adsorption or coupled filtration and adsorption and other types of adsorption using physical or chemical forces rather than driving forces as used normally in hemofiltration derived techniques. The second concept is called the threshold immunomodulation hypothesis, also called the Honore concept. In this concept the view of the system is much more dynamic. In experiments when removal is occurring on the blood compartment side, the level on the interstitial side and the tissue side is also changing and, because not only mediators but also pro-mediators are being removed, some pathways have really stopped when enough pro-mediators have been removed by this technique. At this point, the cascade is blocked and this point is called the threshold point. At this level, the cascade is lost and no further harm can be done to the tissue of the organism. Obviously, it is difficult to know when this point has been reached once high-volume hemofiltration is applied. But what is known, is that hemodynamics and survival can be improved in some patients as shown by various studies using high-volume hemofiltration without any significant drop in mediators inside the blood compartment itself. This effect is obtained without a dramatic fall Honore/Joannes-Boyau/Gressens 388 in the plasma cytokine level because the cytokine or mediator levels should fall at the tissue level and not specifically at the blood compartment level. Nevertheless, the exact mechanism by which high-volume hemofiltration increases the flow of mediators and cytokines between the interstitial compartment and the blood compartment (and back to the blood side) is not known. Before the end of 2005, it was found that this missing step is perhaps well explained by the last theory and/or concept. The third theory and concept is called the mediator delivery hypothesis and has also been called the Alexander concept. In this theory, the use of high-volume hemofiltration and especially high intakes of incoming fluids (3-5 l/h) is able to increase the lymphatic flow 20- to 40-fold, even more so for mediators and cytokine lymphatic flow (drag). This has been demonstrated by several reports and is obviously extremely important. Perhaps this can explain why some very recent studies using high-permeability hemofiltration in sepsis have not been effective in improving hemodynamics and survival in septic acute animal models. In summary various brand new theories will be reviewed here in depth. PMID- 17464150 TI - Liver support systems. AB - Liver insufficiency is a dramatic syndrome with multiple organ involvement. A multiplicity of toxic substances (hydrophilic like ammonia and lipophilic like bilirubin or bile acids or mercaptans) are released into the systemic circulation, thus altering many enzymatic cellular processes. Patients frequently die while on the transplantation waiting list because of organ scarcity. Systems supporting liver function may be useful to avoid further complications due to the typical toxic state, 'bridging' the patients to the transplantation, or, in the event of an acute decompensation of a chronic liver disease, sustain liver function long enough to permit the organ's regeneration and functional recovery. An ideal liver support system should substitute the main functions of the liver (detoxification, synthesis and regulation). Extracorporeal systems now available may be totally artificial or bioartificial. While the first are only able to perform detoxification, the second may add the functions of synthesis (plasma proteins, coagulation factors) and regulation (neurotransmitters). Bioartificial liver working with isolated hepatocytes and a synthetic membrane in an extracorporeal system are however still far from being ready for clinical use. At present, liver insufficiency may be treated with an extracorporeal support technology aimed either at detoxification alone or at a real purification. Charcoal hemoperfusion or exchange/absorption resins may be used for blood detoxification. Blood or plasma exchange, from a theoretical point of view, could be suitable for a polyvalent intoxication, such as liver failure; however, the multicompartmental distribution of some solutes largely endangers the efficacy of these procedures. Selective plasmapheresis techniques are now available for some solutes (e.g. styrene for bilirubin) and may progressively reduce the plasma levels and presumably the deposits of the solute. Novel treatments introduced to improve detoxification, mainly of the protein-bound substances, are the molecular adsorbent recirculation system (MARS) and Prometheus systems. MARS performs an albumin dialysis, where albumin is the exogenous carrier for the toxic substances, and different experiences have proved its efficacy mainly in the treatment of hepatic encephalopathy, while data on survival are still limited to small case series. With Prometheus, the most recent system developed for a wide Liver Support Systems 397 detoxification, albumin-bound toxins are directly removed in two separate cartridges with different solute affinity, without the need for exogenous albumin; plasmadsorption is then coupled with a real dialysis process. After promising initial results, the efficacy of Prometheus in the patients' hard endpoints will be evaluated in a large international trial. On the whole, liver support systems may offer, in many cases, a survival benefit. Stem cells are however, even in this filed, the real great hope for the future of patients with end-stage liver disease. PMID- 17464151 TI - Coupled plasma filtration adsorption. AB - Sepsis is one of the main causes of death in critically ill patients worldwide, and in many cases it is associated with renal and/or other organ failure. However, we do not have a unique efficient therapy to reduce this extremely high mortality rate. In the last years interest around the use of extracorporeal blood purification techniques has increased. One of the emerging treatments in patients with severe sepsis and septic shock is coupled plasma filtration adsorption (CPFA), a novel extracorporeal blood purification therapy aimed at a nonselective reduction of the circulating levels and activities of both pro- and anti inflammatory mediators. Early experimental studies and the following clinical trials have demonstrated impressive results regarding hemodynamics and respiratory parameters, even in patients without concomitant acute renal injury, paralleled by a quick tapering of vasoactive drugs. Considering the still high morbidity and mortality rates in septic shock patients, this new blood purification technique seems to have benefits when applied early in the course of sepsis, also without renal indications, suggesting that it might be performed to prevent rather than to treat acute kidney injury. PMID- 17464152 TI - Albumin dialysis and plasma filtration adsorption dialysis system. AB - Albumin-bound toxins are important in the pathophysiology of liver failure, systemic inflammatory response syndrome, and poisoning. Due to its intrinsic ability to bind molecules, albumin has been used in blood purification techniques, such as single pass albumin dialysis, the molecular adsorbent recirculating system and the Prometheus systems. Plasma filtration adsorption dialysis is the latest technology that can combine the best processes of blood/plasma purification in order to determine a selective and effective purification of molecules implicated in liver failure. PMID- 17464153 TI - Renal assist device and treatment of sepsis-induced acute kidney injury in intensive care units. AB - Acute kidney injury (AKI) is a frequent and serious complication of sepsis in ICU patients and is associated with a very high mortality. Despite the advent of sophisticated renal replacement therapies (RRT) employing high-dose hemofiltration and high-flux membranes, mortality and morbidity from sepsis induced AKI remained high. Moreover, these dialytic modalities could not substitute for the important functions of renal tubular cells in decreasing sepsis-induced AKI biological dysregulations. The results from the in vitro and preclinical animal model studies were very intriguing and led to the development of a bioartificial kidney consisting of a renal tubule assist device containing human proximal tubular cells (RAD) added in tandem to a continuous venovenous hemofiltration circuit. The results from the phase I safety trial and the recent phase II clinical trial showed that the RAD not only can replace many of the indispensable biological kidney functions, but also modify the natural history of sepsis-induced AKI by ameliorating patient survival. PMID- 17464154 TI - Renal replacement therapy in neonates with congenital heart disease. AB - BACKGROUND: The acute renal failure (ARF) incidence in pediatric cardiac surgery intensive care unit (ICU) ranges from 5 to 20% of patients. In particular, clinical features of neonatal ARF are mostly represented by fluid retention, anasarca and only slight creatinine increase; this is the reason why medical strategies to prevent and manage ARF have limited efficacy and early optimization of renal replacement therapy (RRT) plays a key role in the outcome of cardiopathic patients. METHODS: Data on neonates admitted to our ICU were prospectively collected over a 6-month period and analysis of patients with ARF analyzed. Indications for RRT were oligoanuria (urine output less than 0.5 ml/kg/h for more than 4 h) and/or a need for additional ultrafiltration in edematous patients despite aggressive diuretic therapy. RESULTS: Incidence of ARF and need for RRT were equivalent and occurred in 10% of admitted neonates. Eleven patients of 12 were treated by peritoneal dialysis (PD) as only RRT strategy. PD allowed ultrafiltration to range between 5 and 20 ml/h with a negative balance of up to 200 ml over 24 h. Creatinine clearance achieved by PD ranged from 2 to 10 ml/min/1.73 m2. We reported a 16% mortality in RRT patients. CONCLUSION: PD is a safe and adequate strategy to support ARF in neonates with congenital heart disease. Fluid balance control is easily optimized by this therapy whereas solute control reaches acceptable levels. PMID- 17464155 TI - The DOse REsponse Multicentre International Collaborative Initiative (DO-RE-MI). AB - BACKGROUND: Current practices for renal replacement therapy (RRT) in ICU remain poorly defined. The observational DOse REsponse Multicentre International collaborative initiative (DO-RE-MI) survey addresses the issue of how the different modes of RRT are currently chosen and performed. The primary endpoint of DO-RE-MI will be the delivered dose versus in ICU, 28-day, and hospital mortality, and the secondary endpoint, the hemodynamic response to RRT. Here, we report the first preliminary descriptive analysis after 1-year recruitment. METHODS: Data from 431 patients in need of RRT with or without acute renal failure (mean age 61.2+15.9) from 25 centers in 5 countries (Spain, Italy, Germany, Portugal, France) were entered in electronic case report forms (CRFs) available via the website acutevision.net. RESULTS: On admission, 51% patients came from surgery, 36% from the emergency department, and 16% from internal medicine. On admission, mean SOFA and SAPS II were 13 and 50, respectively. The first criteria to initiate RRT was the RIFLE in 38% (failure: 70%, injury: 25%, risk: 22%), the second the high urea/creatinine, and the third immunomodulation. A total of 3,010 cumulative CRF were reported: continuous venovenous hemodiafiltration (CVVHDF) 60%, continuous venovenous hemofiltration (CVVH) 15%, intermittent hemodialysis (IHD) 15%, high-volume hemofiltration (HVHF) 7%, continuous venovenous hemodialysis (CVVHD) 1%, and coupled plasma filtration adsorption/CVVD 2%. In 15% of cases, the patient was shifted to another modality. Mean blood flow rates (ml/min) in the different modalities were: 145 (CVVHDF), 200 (CVVH), 215 (IHD), 283 (HVHF), and 150 (CVVHD). Downtime ranged from 8 to 28% of the total treatment time. Clotting of the circuit accounted for 74% of treatment interruptions. CONCLUSIONS: Despite a large variability in the criteria of choice of RRT, CVVHDF remains the most used (49%). Clotting and clinical reasons were the most common causes for RRT downtime. In continuous RRT, a large variability in the delivered dose is observed in the majority of patients and often in the same patient from one day to another. Preliminary analysis suggests that in a large number of cases the delivered dose is far from the 'adequate' 35 ml/h/kg. PMID- 17464156 TI - Clinical effects of polymyxin B-immobilized fiber column in septic patients. AB - Endotoxin is one of the principal biological substances that cause gram-negative septic shock. Lack of clinical success with antiendotoxin or anticytokine therapy has shifted interest to extracorporeal therapies to reduce circulating levels of the mediators of sepsis. Direct hemoperfusion with polymyxin-B-immobilized fiber (PMX-F) is a promising treatment of gram-negative sepsis in critically ill patients. Because of the high affinity of polymyxin B for endotoxin, the rationale underlying extracorporeal therapy would be to remove circulating endotoxin by adsorption, thus preventing progression of the biological cascade of sepsis. In a systematic review of 28 studies (pooled sample size 1,390 patients), the preliminary results of which are described here, PMX-F therapy appeared to significantly lower endotoxin levels, improve blood pressure, and reduce mortality. However, publication bias and lack of blinding need to be considered. These encouraging results need to be verified with large-scale controlled clinical trials. PMID- 17464157 TI - An updated systematic review of the pharmacology of silymarin. AB - BACKGROUND: Recent years have seen an explosion of scientific papers that deal with drugs from the fruits of milk thistle and its active substances silymarin (standardized mixture of flavonolignanes), thus justifying an updated systematic review. METHODS: Electronic databases identified silymarin, silibinin, silicristin or milk thistle as descriptors in >700 papers (34% published in last 5 years; 92% dealt with animal pharmacological). Only papers adequately reporting on experimental conditions, dosing, variables tested and statistics were analysed. RESULTS: Silymarin was found to modify specifically the functions related to various transporters and receptors located in the cell membranes; that is, organic anion uptake transporter peptides (OATP), ABC transporters (P-gp), bile salt export pump, as well as TNF-alpha-dependent and possibly selectin dependent phenomena. In the cytoplasm, some antioxidant properties and the inhibition of the lipoxygenase pathway seem quite selective and could concur to the antitoxic effects. Some effects like the inhibition of inducible nitric-oxide synthase, of nuclear factor kappa B, and reduction of collagen synthesis are indicative of DNA/RNA-mediated effects. Several studies using 'in vitro' and 'in vivo' cancer models suggest a potential of silymarin in such diseases. Topical and systemic silymarin has skin protective properties against UV-induced damage in epidermis and causes an up-regulation of tumour-suppressor genes p53- and p21CIP1. There were no data on hepatic viral replication, viremia or spontaneous tumours in the data examined. CONCLUSIONS: Data presented here do not solve the question about the complex mechanism(s) of action of the medicinal herbal drug silymarin. Silymarin may be a natural multi-functional and multi-target drug. PMID- 17464158 TI - [Acute effects of needle acupuncture on power performance during stretch shortening cycle]. AB - BACKGROUND: Some studies have shown beneficial effects of needle acupuncture on physical performance. OBJECTIVE: To evaluate immediate effects of a standardized acupuncture treatment on vertical jumping performance. SUBJECTS AND METHODS: 12 healthy sport students participated in the study. A randomized crossover design was used to investigate specific effects of real acupuncture, sham acupuncture and a no-acupuncture waiting list condition on myoelectric activity and kinematic parameters (duration of ground contact, maximum jumping height) in one-legged drop jumps. RESULTS: The results of the present study do not demonstrate significant treatment effects on myoelectric and kinematic parameters. However, real acupuncture tends to result in a relative decrease in the duration of ground contact accompanied by increased muscular innervation. CONCLUSION: Acupuncture treatment had no significant impact on muscular performance of lower leg in stretch-shortening cycle under the present conditions. As to further research, the potential efficacy of acupuncture for improving reactive strength should be investigated in terms of controlled trials with stratified randomization according to physical performance capacity. PMID- 17464159 TI - [Pilot study of breathing therapy in groups for patients with bronchial asthma]. AB - BACKGROUND: The present study examines the efficacy of breathing and body therapy (according to the organization of breathing therapists, BVA) on bronchial asthma. This psychophysical therapy procedure has never before been studied for its efficacy on asthma, much in contrast to breathing exercises, which have been extensively examined. PATIENTS AND METHODS: This uncontrolled observational study is of preliminary character. It examines the influence of psychophysical breathing and body exercises practiced in short-term intervention groups on quality of life (SF-36) as well as subjective discomfort (discomfort diary) in patients with bronchial asthma. As there are no prior studies on the breathing and body therapy (BVA) method, it was hypothesized, based on experience, that it would improve ventilation, lower anxiety, and perhaps even allow for a reduction of medication. RESULTS: An anti-obstructive effect could not be addressed here, as the study only included patients who were free of clinically relevant obstructions through conventional treatment. However, a tendency for an anxiety relieving effect could be observed in the SF-36. Subjective reports of respiratory discomfort, obtained through assessment of the patients' diaries, did not show any tendencies or even significant changes over the course of the therapy. CONCLUSION: The suggested anxiety-relieving effect of breathing therapy should be examined in future studies with more patients and stricter inclusion criteria. PMID- 17464160 TI - Prophylactic role for complementary and alternative medicine in perinatal programming of adult health. AB - BACKGROUND: The health status of an individual in adulthood is proposed to be determined by events occurring in the prenatal and early postnatal period. A common early life event proven to have long lasting effects on the developing fetus is stress, including pain. Exposure of fetal and neonatal infants to repetitive psychological (e.g., maternal stress) or physiological (e.g., pain, infection, and noise) stress during this period is proposed to alter mechanisms involved in the regulation of stress, immunological maturation, pain perception, and cognition. Such changes, which persist into adulthood, may occur via alterations in the development of the hypothalamic-pituitary-adrenal (HPA) axis. This process is typically referred to as 'perinatal programming'. Ontogenic alterations in the development of the HPA-axis have been related to a number of adult pathologies such as cardiovascular disease, type 2 diabetes, asthma, as well as psychopathologies such as anxiety and depression. OBJECTIVE: In this review, the effectiveness of complementary and alternative medicine (CAM), such as music, dietary supplements, massage and aromatherapy, in reducing perinatal stress in mothers and infants is examined. An emphasis is placed on these therapies as preventative measures which may be of value to individuals at risk of developing disease profiles associated with the consequences of adverse perinatal programming. The widening interest in perinatal programming and CAM suggests the potential for CAM to become a valuable tool in offsetting negative adult health outcomes resulting from perinatal programming associated with adverse gestational early life environments. PMID- 17464161 TI - Evidence based complementary and alternative medicine: promises and problems. AB - OBJECTIVE: The present paper examines the experience of establishing a center for evidence-based complementary and alternative medicine (EBCAM) practice. It examines both the difficulties and the challenges of doing research to establish EBCAM. The paper also examines the political context of the demand for evidence- based practice (EBP) for CAM. IMPLEMENTATION: A center for EBCAM was funded for 3 years within the Southern California Evidence-Based Practice Center by the National Center for CAM and administered by the Agency for Health Research Quality. This experience provides the basis for this paper. RESULTS: While the experience of creating an EBM Center for CAM has shown that much work can be accomplished by applying standard methods of EBP medicine, it also highlights the weaknesses of such an agenda. Many standard research methods are simply not applicable to CAM, and even where they are, effectiveness is a much more important means of assessing CAM than simply efficacy. Researchers however, must be conscious of the political motivations behind much of the demand for EBCAM. Where such demands are coming from allopathic medicine, they clearly form a continuing part of medical opposition to CAM and may be intended to perpetuate the dominance of the biomedical paradigm in healthcare. The challenge for CAM is to recognize the limitations of EBP but not to throw the 'baby out with the bathwater'. There is much in EBP that clearly should be emulated by the CAM community but only where it is appropriate. PMID- 17464162 TI - Is black cohosh a hepatotoxic medicinal herb? PMID- 17464164 TI - [Enterovesical fistula and intestinal perforation complicated by Crohn's disease]. PMID- 17464165 TI - [Prevention of hepatocellular carcinoma]. AB - Hepatocellular carcinoma (HCC) is one of the most common malignant tumors and has the third highest mortality rate among malignancies in South Korea. Despite the continuing efforts for the early detection of HCC, the mortality rate and prognosis have not been improved yet. Its clinical behavior is quite different from other cancers. High recurrence rate after curative treatment might be the reason for poor prognosis. Several methods including chemoprevention, blocking the development of HCC, have been under investigations. The vaccine for hepatitis, in the form of primary prevention, is considered to be the most effective one inhibiting the development of liver disease. Furthermore, keeping away from hepatotoxic agents is another way for preventing liver cell injuries. Secondary prevention is to stop the development of HCC in chronic liver diseases. Since the level of DNA in hepatitis B virus (HBV) hepatitis patients is closely related with the development of HCC, it is helpful to lower the DNA level using anti-viral agents. In addition, IFN, one of the anti-viral agents, can inhibit HCV hepatitis from tumorigenesis. Cyclo-oxygenase (COX)-2 inhibitors are also alleged to have a function in interrupting the development of HCC. Tertiary prevention means the prevention of recurrence of HCC after successful treatment. Because of high recurrence rate, the prevention of recurrence should be one of the important factors affecting the prognosis of HCC. Up to now, COX inhibitors, retinoic acids, vitamin K2, glycyrrhizin epigallocatechin-3-gallate (EGCG), and ginseng had been reported to be effective for the chemoprevention of HCC. Further studies are required for an advancement in the prevention of HCC. PMID- 17464166 TI - [Gene expression profiling using oligonucleotide microarray in atrophic gastritis and intestinal metaplasia]. AB - BACKGROUND/AIMS: The atrophic gastritis with intestinal metaplasia of gastric mucosa has been considered to be the major factor of carcinogenesis in the stomach. However, the key molecules are still poorly understood. To elucidate the molecular genetic basis, we report the results of our initial microarray data to analyze the genome pattern in patients with atrophic gastritis and intestinal metaplasia of the stomach. METHODS: We used oligonucleotide microarray technique to evaluate the gene expression profiles in atrophic gastritis with intestinal metaplasia, in comparison with those of normal mucosa. For the identification of differentially expressed genes, Significance Analysis of Microarrays (SAM) package method was used. The results were analyzed using global normalization, intensity dependent normalization, and box plot normalization. RESULTS: Eight genes including FABP, REG, OR6C1, MEP1, SLC6A1, SI, Mucin 1, and RAB23 in mucosa of atrophic gastritis and intestinal metaplasia were up-regulated by more than 10 times as compared with normal gastric mucosa. Only one gene, LOC44119 was down regulated by more than 10 times of the expression as compared with normal gastric mucosa. In respect to the expression of known genes related to gastric carcinogenesis, 8 genes including FN1, SRMS, TP53, TP53IMP2, TP53I3, FGFR4, TGFB1, and TGFA showed up- and down-regulations more than 2 folds in expression pattern. CONCLUSIONS: We could identify a total genome pattern in patient with atrophic gastritis and intestinal metaplasia using oligonucleotide microarray. We believe that the current results will serve as a fundamental bioinformative basis for clinical applications in diagnosis and treatment of gastric cancer and precancerous lesion in the future. PMID- 17464167 TI - [Contribution of nasal methicillin-resistant Staphylococcus aureus colonization to percutaneous endoscopic gastrostomy site infection and risk factors of wound infection]. AB - BACKGROUND/AIMS: Peristomal infection is the most common complication of percutaneous endoscopic gastrostomy (PEG) insertion. Methicillin-resistant Staphylococcus aureus (MRSA) is the most commonly implicated organism of peristomal infection. The aims of this study were to determine the contribution of nasal MRSA to wound infection in PEG and the predictors of wound infection. METHODS: A prospective study was conducted on patients undergoing PEG between September 2003 and July 2005. All patients received antibiotics prior to PEG insertion. Nasal swabs were taken from a consecutive series of patients prior to PEG insertion. Wound status of the peristomal site were prospectively evaluated at day 1, 3, and 7 following the insertion of PEG. RESULTS: Thirty-one patients underwent PEG insertion (mean age, 66+/-16 years). Ten patients (32.3%) had MRSA positive nasal swabs. Peristomal infection did not have any relationship with nasal MRSA colonization (p>0.05). Peristomal infection occurred in 4 (12.9%) cases. The rate of peristomal infections was significantly higher in patients with diabetes mellitus (p<0.05). CONCLUSIONS: Nasal MRSA colonization is not associated with the risk of peristomal infections in patients receiving antibiotics prior to PEG insertion. Diabetes mellitus might be the risk factor for peristomal infection after PEG insertion. PMID- 17464168 TI - [An evaluation of web-based informations about gastroesophageal reflux diseases in Korea]. AB - BACKGROUND/AIMS: Internet has become an important source of medical information not only for medical personnels but also for patients. The aim of this study was to evaluate the quality of internet based medical information about 'gastroesophageal reflux' or 'reflux esophagitis' in Korea. METHODS: The first 15 internet sites using the key words 'gastroesophageal reflux' or 'reflux esophagitis' were retrieved from the 7 most frequently used internet search engines. The quality of information from a total of 108 websites was evaluated using a checklist. RESULTS: Among total 108 sites related to 'gastroesophageal reflux' or 'reflux esophagitis', fifty-six sites (51.8%) were made by hospitals or clinics and 94 sites (87.0%) were made for patients. Of the 108 sites, eleven web sites (10.1%) had more than three JAMA benchmarks (authorship, references, currency, and disclosure). Higher quality sites (at least three JAMA benchmarks) were less likely to contain inaccurate information than lower quality sites (fewer than three JAMA benchmarks)-3/11 (27.2%) vs. 60/97 (61.9%) (p<0.01). Despite the fact that articles in the literature emphasized an insufficient evidence to support an association between the lifestyle, dietary behaviors, and GERD, such guidelines continue to be recommended as first-line therapy in most websites. CONCLUSIONS: Informations about gastroesophageal reflux disease were incomplete in the majority of medical web sites. These would bring about confusion to patients seeking for an information about GERD through the internet. There is a need for better sources in evidence based informations about gastroesophageal reflux diseases on the web. PMID- 17464169 TI - [Comparison of clinical characteristics between cryptogenic and biliary pyogenic liver abscess]. AB - BACKGROUND/AIMS: The aim of this study was to evaluate the characteristics of cryptogenic and biliary pyogenic liver abscess by comparing the clinical aspects between the two groups. METHODS: Of 119 patients treated for pyogenic liver abscess from 2000 to 2004, 82 subjects with cryptogenic liver abscess and 21 with biliary abscess were analysed retrospectively. RESULTS: There were no significant differences between the two groups regarding clinical symptoms. The characteristics of abscess were similar except the size of abscess. The size of abscess less than 5 cm in diameter was more common in biliary group than in cryptogenic group (p=0.004). Compare to cryptogenic group, biliary group had more positive culture test from abscess (100% vs. 69%, p=0.006). Especially, E. coli isolated from abscess culture (28% vs. 7%, p=0.014) and blood culture (23% vs. 4%, p=0.035) were more common in biliary group than in cryptogenic group. There was no difference in mortality between the two groups (biliary vs. cryptogenic: 4.8% vs. 0%, p=0.204). However, the rate of clinical improvement was higher in cryptogenic group than in biliary group (92.7% vs. 66.7%, p=0.001). CONCLUSIONS: Biliary liver abscess had similar clinical characteristics to cryptogenic origin. Biliary liver abscess had smaller abscess size and more positive abscess culture rates than cryptogenic abscess. Improvement rate without complication and recurrence was higher in cryptogenic group than biliary group. PMID- 17464170 TI - Metachronous gastric MALT lymphoma and early gastric cancer: a case report. AB - Metachronous association between gastric lymphoma and early gastric cancer is a rare event. Recent studies have suggested that a relationship exists between gastric mucosa-associated lymphoid tissue (MALT) lymphoma and gastric carcinoma although the mechanism is unknown. Herein, we report a 53-year-old man who visited to our hospital due to melena. Esophagogastroduodenoscopy (EGD) revealed a MALT lymphoma on the greater curvature of lower body. The patient received anti Helicobacter pylori eradication therapy, followed by 6 cycles of chemotherapy and radiation therapy, and achieved complete remission 12 months after the therapy. Three years later, he revisited our hospital with epigastric pain. EGD revealed an early gastric cancer on the anterior wall of proximal antrum, nearly opposite to the previous lymphoma site, and a partial gastrectomy was performed. To the best of our knowledge, this is the first case report of metachronous MALT lymphoma and subsequent gastric carcinoma in Korea. PMID- 17464171 TI - [A case with Rotor syndrome in hyperbilirubinemic family]. AB - Rotor syndrome is a rare, benign familial disorder characterized by chronic fluctuating, nonhemolytic and predominantly conjugated hyperbilirubinemia with normal hepatic histology. In contrast to Dubin-Johnson syndrome, there is no liver pigmentation in Rotor syndrome. A 36-year-old man was admitted due to asymptomatic persistent jaundice. His siblings had jaundice with direct hyperbilirubinemia. Physical examination revealed icteric sclerae without hepatosplenomegaly. Laboratory findings showed increased serum bilirubin with direct bilirubinemia. Hepatic uptake and storage capacity of indocyanine green was markedly reduced, while excretion into bile was slightly suppressed. Markedly decreased hepatic uptake and poor visualization of the gallbladder and biliary tract were shown in 99mTc-DISIDA scan. Histology of the liver showed mild steatosis without pigmentation. We report a case with the review of literature. PMID- 17464172 TI - [Intussusception secondary to jejunal lymphangioma in a 13-year-old girl]. AB - Lymphangioma is a benign tumor composed of numerous small or large thin-walled lymphatic spaces. It shows a predilection for head, neck, and axilla. Jejunal lymphangioma in children is extremely rare. We present a case of 13-year-old girl with jejuno-jejunal intussusception secondary to lymphangioma in jejunum. To our knowledge, this is the second report of jejunal lymphangioma causing intussusception in a child. PMID- 17464173 TI - [Microarray analysis of multiple gene expression in intestinal metaplasia and atrophic gastritis]. PMID- 17464174 TI - p38 MAPK and ERK activation by 9-cis-retinoic acid induces chemokine receptors CCR1 and CCR2 expression in human monocytic THP-1 cells. AB - 9-cis-Retinoic acid (9CRA) plays an important role in the immune response; this includes cytokine production and cell migration. We have previously demonstrated that 9CRA increases expression of chemokine receptors CCR1 and CCR2 in human monocytes. To better understand how 9CRA induces CCR1 and CCR2 expression, we examined the contribution of signaling proteins in human monocytic THP-1 cells. The mRNA and surface protein up-regulation of CCR1 and CCR2 in 9CRA-stimulated cells were weakly blocked by the pretreatment of SB202190, a p38 MAPK inhibitor, and PD98059, an upstream ERK inhibitor. Activation of p38 MAPK and ERK1/2 was induced in both a time and dose-dependent manner after 9CRA stimulation. Both p38 MAPK and ERK1/2 phosphorylation peaked at 2 h after a 100 nM 9CRA treatment. 9CRA increased calcium influx and chemotactic activity in response to CCR1-dependent chemokines, Lkn-1/CCL15, MIP-1alpha/CCL3, and RANTES/CCL5, and the CCR2-specific chemokine, MCP-1/CCL2. Both SB202190 and PD98059 pretreatment diminished the increased calcium mobilization and chemotactic ability due to 9CRA. SB202190 inhibited the expression and functional activities of CCR1 and CCR2 more effectively than did PD98059. Therefore, our results demonstrate that 9CRA transduces the signal through p38 MAPK and ERK1/2 for CCR1 and CCR2 up regulation, and may regulate the pro-inflammatory process through the p38 MAPK and ERK-dependent signaling pathways. PMID- 17464175 TI - Curcumin reduces cold storage-induced damage in human cardiac myoblasts. AB - Curcumin is a polyphenolic compound possessing interesting anti-inflammatory and antioxidant properties and has the ability to induce the defensive protein heme oxygenase-1 (HO-1). The objective of this study was to investigate whether curcumin protects against cold storage-mediated damage of human adult atrial myoblast cells (Girardi cells) and to assess the potential involvement of HO-1 in this process. Girardi cells were exposed to either normothermic or hypothermic conditions in Celsior preservation solution in the presence or absence of curcumin. HO-1 protein expression and heme oxygenase activity as well as cellular damage were assessed after cold storage or cold storage followed by re-warming. In additional experiments, an inhibitor of heme oxygenase activity (tin protoporphyrin IX, 10 microM) or siRNA for HO-1 were used to investigate the participation of HO-1 as a mediator of curcumin-induced effects. Treatment with curcumin produced a marked induction of cardiac HO-1 in normothermic condition but cells were less responsive to the polyphenolic compound at low temperature. Cold storage-induced damage was markedly reduced in the presence of curcumin and HO-1 contributed to some extent to this effect. Thus, curcumin added to Celsior preservation solution effectively prevents the damage caused by cold-storage; this effect involves the protective enzyme HO-1 but also other not yet identified mechanisms. PMID- 17464176 TI - Coptidis rhizoma extract protects against cytokine-induced death of pancreatic beta-cells through suppression of NF-kappaB activation. AB - We demonstrated previously that Coptidis rhizoma extract (CRE) prevented S nitroso-N-acetylpenicillamine-induced apoptotic cell death via the inhibition of mitochondrial membrane potential disruption and cytochrome c release in RINm5F (RIN) rat insulinoma cells. In this study, the preventive effects of CRE against cytokine-induced beta-cell death was assessed. Cytokines generated by immune cells infiltrating pancreatic islets are crucial mediators of beta-cell destruction in insulin-dependent diabetes mellitus. The treatment of RIN cells with IL-1beta and IFN-gamma resulted in a reduction of cell viability. CRE completely protected IL-1beta and IFN-gamma-mediated cell death in a concentration-dependent manner. Incubation with CRE induced a significant suppression of IL-1beta and IFN-gamma-induced nitric oxide (NO) production, a finding which correlated well with reduced levels of the iNOS mRNA and protein. The molecular mechanism by which CRE inhibited iNOS gene expression appeared to involve the inhibition of NF-kappaB activation. The IL-1beta and IFN-gamma stimulated RIN cells showed increases in NF-kappaB binding activity and p65 subunit levels in nucleus, and IkappaB alpha degradation in cytosol compared to unstimulated cells. Furthermore, the protective effects of CRE were verified via the observation of reduced NO generation and iNOS expression, and normal insulin secretion responses to glucose in IL-1beta and IFN-gamma-treated islets. PMID- 17464177 TI - A point mutant of apolipoprotein A-I, V156K, exhibited potent anti-oxidant and anti-atherosclerotic activity in hypercholesterolemic C57BL/6 mice. AB - In our previous study, two point mutants of apolipoprotein A-I, designated V156K and A158E, revealed peculiar characteristics in their lipid-free and lipid-bound states. In order to determine the putative therapeutic potential of these mutants, several in vitro and in vivo evaluations were conducted. In the lipid free state, V156K showed more profound antioxidant activity against LDL oxidation than did the wildtype (WT) or A158E variants in an in vitro assay. In the lipid bound state, V156K-rHDL showed an enhanced cholesterol delivery activity to HepG2 cells in a time-dependent manner, as compared to WT-rHDL, A158E-rHDL, and R173C rHDL. We assessed the physiological activities of the mutants in circulation, using hypercholesterolemic mice (C57BL6/J). Palmitoyloleoyl phosphatidylcholine (POPC)-rHDL preparations containing each of the apoA-I variants were injected into the mice at a dosage of 30 mg of apoA-I/kg of body weight. Forty eight hours after injection, the sera of the V156K-rHDL injected group showed the most potent antioxidant abilities in the ferric acid removal assay. The V156K-rHDL- or R173C rHDL-injected mice showed no atherosclerotic lesions and manifested striking increases in their serum apo-E levels, as compared to the mice injected with WT rHDL or A158E-rHDL. In conclusion, V156K-rHDL exhibited the most pronounced antioxidant activity and anti-atherosclerotic activity, both in vitro and in vivo. These results support the notion that HDL-therapy may prove beneficial due to its capacity to induce accelerated cholesterol excretion, as well as its enhanced antioxidant and anti-inflammatory effects and lesion regression effect. PMID- 17464178 TI - Promoter effects of adeno-associated viral vector for transgene expression in the cochlea in vivo. AB - The aims of this study were to evaluate the expression of enhanced green fluorescent protein (EGFP) driven by 6 different promoters, including cytomegalovirus IE enhancer and chicken beta-actin promoter (CAG), cytomegalovirus promoter (CMV), neuron-specific enolase promoter (NSE), myosin 7A promoter (Myo), elongation factor 1alpha promoter (EF-1alpha), and Rous sarcoma virus promoter (RSV), and assess the dose response of CAG promoter to transgene expression in the cochlea. Serotype 1 adeno-associated virus (AAV1) vectors with various constructs were transduced into the cochleae, and the level of EGFP expression was examined. We found the highest EGFP expression in the inner hair cells and other cochlear cells when CAG promoter was used. The CMV and NSE promoter drove the higher EGFP expression, but only a marginal activity was observed in EF-1alpha promoter driven constructs. RSV promoter failed to driven the EGFP expression. Myo promoter driven EGFP was exclusively expressed in the inner hair cells of the cochlea. When driven by CAG promoter, reporter gene expression was detected in inner hair cells at a dose as low as 3x10(7) genome copies, and continued to increase in a dose-dependent manner. Our data showed that individual promoter has different ability to drive reporter gene expression in the cochlear cells. Our results might provide important information with regard to the role of promoters in regulating transgene expression and for the proper design of vectors for gene expression and gene therapy. PMID- 17464179 TI - CD99 activates T cells via a costimulatory function that promotes raft association of TCR complex and tyrosine phosphorylation of TCR zeta. AB - We investigated the co-stimulatory role of a cell-surface protein, CD99. Co ligation of CD99 and suboptimal CD3 induced T-cell activation to a level comparable to that obtained with optimal CD3 or CD3+CD28. We also noted concomitant enhancement of the earliest T-cell receptor (TCR) signaling events. In addition, co-ligation of CD99 and CD3 led to translocation of TCR complexes into the lipid raft, without concomitant migration of CD99 to the raft, and consequent enhancement of TCR zeta-mediated signal 1. These data demonstrate the unique properties of CD99 co-stimulation that distinguish this molecule from CD28 and other raft-resident co-stimulatory factors. PMID- 17464180 TI - Phytosphingosine-1-phosphate stimulates chemotactic migration of L2071 mouse fibroblasts via pertussis toxin-sensitive G-proteins. AB - Phytosphingosine-1-phosphate (PhS1P) was found to stimulate an intracellular calcium increase via phospholipase C but not pertussis toxin (PTX)-sensitive G proteins in L2071 mouse fibroblasts. PhS1P also activated ERK and p38 kinase, and these activations by PhS1P were inhibited by PTX. Moreover, PhS1P stimulated the chemotactic migration of L2071 cells via PTX-sensitive Gi protein(s). In addition, the PhS1P-induced chemotactic migration of L2071 cells was also dramatically inhibited by LY294002 and SB203580 (inhibitors of phosphoinositide 3 kinase and p38 kinase, respectively). L2071 cells are known to express four S1P receptors, i.e., S1P1, S1P2, S1P3, and S1P4, and pretreatment with an S1P1 and S1P3 antagonist (VPC 23019) did not affect on PhS1P-induced chemotaxis. This study demonstrates that PhS1P stimulates at least two different signaling cascades, one is a PTX-insensitive but phospholipase C dependent intracellular calcium increase, and the other is a PTX-sensitive chemotactic migration mediated by phosphoinositide 3-kinase and p38 kinase. PMID- 17464181 TI - Differential promoter methylation may be a key molecular mechanism in regulating BubR1 expression in cancer cells. AB - The BubR1 mitotic-checkpoint protein monitors proper attachment of microtubules to kinetochores, and links regulation of chromosome-spindle attachment to mitotic checkpoint signaling. Thus, disruption of BubR1 activity results in a loss of checkpoint control, chromosomal instability caused by a premature anaphase, and/or the early onset of tumorigenesis. The mechanisms by which deregulation and/or abnormalities of BubR1 expression operate, however, remain to be elucidated. In this study, we demonstrate that levels of BubR1 expression are significantly increased by demethylation. Bisulfite sequencing analysis revealed that the methylation status of two CpG sites in the essential BubR1 promoter appear to be associated with BubR1 expression levels. Associations of MBD2 and HDAC1 with the BubR1 promoter were significantly relieved by addition of 5-aza-2' deoxycytidine, an irreversible DNA methyltransferase inhibitor. However, genomic DNA isolated from 31 patients with colorectal carcinomas exhibited a +84A/G polymorphic change in approximately 60% of patients, but this polymorphism had no effect on promoter activity. Our findings indicate that differential regulation of BubR1 expression is associated with changes in BubR1 promoter hypermethylation patterns, but not with promoter polymorphisms, thus providing a novel insight into the molecular regulation of BubR1 expression in human cancer cells. PMID- 17464182 TI - Identification of novel substrates for human checkpoint kinase Chk1 and Chk2 through genome-wide screening using a consensus Chk phosphorylation motif. AB - Checkpoint kinase 1 (Chk1) and Chk2 are effector kinases in the cellular DNA damage response and impairment of their function is closely related to tumorigenesis. Previous studies revealed several substrate proteins of Chk1 and Chk2, but identification of additional targets is still important in order to understand their tumor suppressor functions. In this study, we screened novel substrates for Chk1 and Chk2 using substrate target motifs determined previously by an oriented peptide library approach. The potential candidates were selected by genome-wide peptide database searches and were examined by in vitro kinase assays. ST5, HDAC5, PGC-1alpha, PP2A PR130, FANCG, GATA3, cyclin G, Rad51D and MAD1a were newly identified as in vitro substrates for Chk1 and/or Chk2. Among these, HDAC5 and PGC-1a were further analyzed to substantiate the screening results. Immunoprecipitation kinase assay of full-length proteins and site directed mutagenesis analysis of the target motifs demonstrated that HDAC5 and PGC-1alpha were specific targets for Chk1 and/or Chk2 at least in vitro. PMID- 17464183 TI - Trichostatin A-mediated upregulation of p21(WAF1) contributes to osteoclast apoptosis. AB - Histone deacetylase inhibitors (HDIs), a new class of anti-cancer agents, have been reported to suppress formation of osteoclast precursors and their fusion into multinucleated cells. However, little is known about the effect of HDIs on mature osteoclasts, which may have significance for their therapeutic use. Here, we demonstrate a novel action of HDIs on osteoclast apoptosis. Primary multinucleated mature osteoclasts were prepared from mouse bone marrow cells. Treatment of osteoclasts with the HDI trichostatin A (TSA) caused apoptosis, as confirmed by annexin V staining and caspase activation. TSA caused the upregulation of p21WAF1 in osteoclasts. To understand the role of p21(WAF1) upregulation in TSA-treated osteoclasts, shRNA against p21(WAF1)-containing lentivirus was introduced into osteoclasts. The suppression of p21(WAF1) decreased TSA-directed osteoclast apoptosis. Collectively, our results provide evidence that TSA causes osteoclast apoptosis, which involves, in part, TSA induced upregulation of p21(WAF1), and strongly supports HDIs as potential therapeutic agents for excessive bone resorption. PMID- 17464184 TI - Resveratrol stimulates glucose transport in C2C12 myotubes by activating AMP activated protein kinase. AB - trans-Resveratrol (t-RVT), a naturally occurring polyphenol found in Polygonum cuspidatum, grape, and red wine, has been reported to have anti-inflammatory, cardioprotective, and cancer chemopreventive properties. However antidiabetic effect of t-RVT has not yet been reported. In this study, we show that t-RVT increases glucose uptake in C2C12 myotubes by activating AMP-activated protein kinase (AMPK), uncovering an antidiabetic potential of t-RVT for the first time. AMPK plays a central role in the regulation of glucose and lipid metabolism, and hence it is considered a novel therapeutic target for metabolic syndrome such as type 2 diabetes. t-RVT significantly induced glucose uptake in C2C12 cells, via AMPK activation, but not a phosphatidylinositol-3 kinase (PI-3 kinase) signal pathway. The induced glucose uptake was attenuated by pretreatment with a pharmacological inhibitor for AMPK, indicating that the effect of t-RVT primarily depends on AMPK activation. However, in the presence of insulin, t-RVT also potentiated the effect of insulin on glucose uptake via AMPK activation, which led to further activation of PI-3 kinase/Akt signal pathway. PMID- 17464185 TI - Colchicine-derived compound CT20126 promotes skin allograft survival by regulating the balance of Th1 and Th2 cytokine production. AB - Colchicine has been shown to regulate the expression of inflammatory gene, but this compound possesses much weaker anti-inflammatory activity. In this study, we synthesized a new colchicine derivative CT20126 and examined its immunomodulatory property. CT20126 was found to have immunosuppressive effects by inhibiting lymphocyte proliferation without cytotoxicity and effectively inhibit the transcriptional expression of the inflammatory genes, iNOS, TNF-alpha, and IL 1beta, in macrophages stimulated by LPS. This effect was nearly comparable to that of cyclosporine A. This compound also significantly suppressed the production of nitric oxide and Th1-related pro-inflammatory cytokines, IL-1beta, TNF-alpha, and IL-2, with minimal suppression of Th2-related anti-inflammatory cytokines IL-4 and IL-10 in the sponge matrix allograft model. Moreover, administration of CT20126 prolonged the survival of allograft skins from BALB/c mice (H-2(d)) to the dorsum of C57BL/6 (H-2(b)) mice. The in vivo immune suppressive effects of CT20126 were similar to that of cyclosporine A. These results indicate that this compound may have potential therapeutic value for transplantation rejection and other inflammatory diseases. PMID- 17464186 TI - Toll-like receptor 9 dependent activation of MAPK and NF-kB is required for the CpG ODN-induced matrix metalloproteinase-9 expression. AB - Unmethylated CpG oligodeoxynucleotides (CpG ODNs) activate immune cells to produce immune mediators. This study demonstrates that in murine macrophage RAW 264.7 cells, CpG ODN-mediated matrix metalloproteinase-9 (MMP-9) expression is regulated at transcriptional level and requires de novo protein synthesis. Inhibition of ERK and p38 MAPK, but not JNK, results in significant decrease of CpG ODN-induced MMP-9 expression. We found that endosomal maturation inhibitors, chloroquine and bafilomycin A, block CpG ODN-induced ERK and p38 MAPK activation and the subsequent MMP-9 expression. We also observed that CpG ODN induces NF kappaB activation and NF-kappaB is a downstream target of p38 MAPK. Taken together, our data demonstrate that CpG ODN triggers MMP-9 expression via TLR-9 dependent ERK and p38 MAPK activation followed by NF-kappaB activation. PMID- 17464187 TI - Heat shock responses for understanding diseases of protein denaturation. AB - Extracellular stresses induce heat shock response and render cells resistant to lethal stresses. Heat shock response involves induction of heat shock proteins (Hsps). Recently the roles of Hsps in neurodegenerative diseases and cancer are attracting increasing attention and have accelerated the study of heat shock response mechanism. This review focuses on the stress sensing steps, molecules involved in Hsps production, diseases related to Hsp malfunctions, and the potential of proteomics as a tool for understanding the complex signaling pathways relevant to these events. PMID- 17464188 TI - The present status of cell tracking methods in animal models using magnetic resonance imaging technology. AB - With the advance of stem cell transplantation research, in vivo cell tracking techniques have become increasingly important in recent years. Magnetic resonance imaging (MRI) may provide a unique tool for non-invasive tracking of transplanted cells. Since the initial findings on the stem cell migration by MRI several years ago, there have been numerous studies using various animal models, notably in heart or brain disease models. In order to develop more reliable and clinically applicable methodologies, multiple aspects should be taken into consideration. In this review, we will summarize the current status and future perspectives of in vivo cell tracking technologies using MRI. In particular, use of different MR contrast agents and their detection methods using MRI will be described in much detail. In addition, various cell labeling methods to increase the sensitivity of signals will be extensively discussed. We will also review several key experiments, in which MRI techniques were utilized to detect the presence and/or migration of transplanted stem cells in various animal models. Finally, we will discuss the current problems and future directions of cell tracking methods using MRI. PMID- 17464189 TI - Lactosylceramide alpha2,3-sialyltransferase is induced via a PKC/ERK/CREB dependent pathway in K562 human leukemia cells. AB - Previously we showed that the human GM3 synthase gene was expressed during the induction of megakaryocytic differentiation in human leukemia K562 cells by phorbol 12-myristate 13-acetate (PMA). In this study we found that treatment of PMA-induced K562 cells with Go6976, a specific inhibitor of PKC, and U0126, an inhibitor of the extracellular signal-regulated kinase (ERK) reduced expression of GM3 synthase, whereas wortmannin, an inhibitor of phosphoinositide 3-kinase (PI3K) did not. Moreover, activation of ERK and cAMP response element binding protein (CREB) was prevented by pretreatment with Go6976 and U0126. PMA stimulated the promoter activity of the 5'-flanking region from -177 to -83 region of the GM3 synthase gene, and mutation or deletion of a CREB site located around -143 of the promoter reduced PMA-stimulated promoter activity, as did the inhibitors Go6976 and U0126. Our results demonstrate that induction of GM3 synthase during megakaryocytic differentiation in PMA-stimulated human leukemia K562 cells depends upon the PKC/ERK/CREB pathway. PMID- 17464190 TI - Isolation of circadian-associated genes in Brassica rapa by comparative genomics with Arabidopsis thaliana. AB - Elucidation of the roles of circadian associated factors requires a better understanding of the molecular mechanisms of circadian rhythms, control of flowering time through photoperiodic pathways, and photosensory signal transduction. In Arabidopsis, the APRR1 quintet, APRRs 1, 3, 5, 7, and 9, are known as central oscillator genes. Other plants may share the molecular mechanism underlying the circadian rhythm. To identify and characterize these circadian response genes in Brassica crops whose genome was triplicated after divergence from Arabidopsis, we identified B. rapa BAC clones containing these genes by BLAST analysis of B. rapa BAC end sequences against the five corresponding Arabidopsis regions. Subsequent fingerprinting, Southern hybridization, and PCR allowed identification of five BAC clones, one for each of the five circadian related genes. By draft shotgun sequencing of the BAC clones, we identified the complete gene sequences and cloned the five expressed B. rapa circadian associated gene members, BrPRRs 1, 3, 5, 7, and 9. Phylogenetic analysis revealed that each BrPRR was orthologous to the corresponding APRR at the sequence level. Northern hybridization revealed that the five genes were transcribed at distinct points in the 24 hour period, and Southern hybridization revealed that they are present in 2, 1, 2, 2, and 1 copies, respectively in the B. rapa genome, which was triplicated and then diploidized during the last 15 million years. PMID- 17464191 TI - Comparative proteomic analysis of blue light signaling components in the Arabidopsis cryptochrome 1 mutant. AB - An Arabidopsis hy4 mutant that is specifically impaired in its ability to undergo blue light dependent photomorphogenesis was used to identify cryptochrome 1 signaling-related components. Proteomic analysis revealed about 205 differentially expressed protein spots in the blue light-irradiated hy4 mutant compared to the wild-type. The proteins corresponding to 28 up-regulated and 33 down-regulated spots were identified. Obvious morphological changes in the hy4 mutant were closely related to the expression of various transcription factors. Our findings suggest that blue light signals may be involved in many cellular processes including disease resistance and stress responses. PMID- 17464192 TI - Molecular and functional characterization of monocot-specific Pex5p splicing variants, using OsPex5pL and OsPex5pS from rice (Oryza sativa). AB - We identified two alternatively spliced variants of the peroxisomal targeting signal 1 (PTS1) receptor protein Pex5ps in monocot (rice, wheat, and barley) but not in dicot (Arabidopsis and tobacco) plants. We characterized the molecular and functional differences between the rice (Oryza sativa) Pex5 splicing variants OsPex5pL and OsPex5pS. There is only a single-copy of OsPEX5 in the rice genome and RT-PCR analysis points to alternative splicing of the transcripts. Putative light-responsive cis-elements were identified in the 5' region flanking OsPEX5L and Northern blot analysis demonstrated that this region affected light-dependent expression of OsPEX5 transcription. Using the pex5-deficient yeast mutant Scpex5, we showed that OsPex5pL and OsPex5pS are able to restore translocation of a model PTS1 protein (GFP-SKL) into peroxisomes. OsPex5pL and OsPex5pS formed homo complexes via specific interaction domains, and interacted with each other and OsPex14p to form hetero-complexes. Although overexpression of OsPex5pL in the Arabidopsis pex5 mutant (Atpex5) rescued the mutant phenotype, overexpression of OsPex5pS only resulted in partial recovery. PMID- 17464193 TI - Identification of novel regulators of apoptosis using a high-throughput cell based screen. AB - High-throughput subcellular imaging is a powerful tool for investigating the function of genes. In order to identify novel regulators of apoptosis we transiently transfected HeLa cells with 938 hypothetical genes of unknown function, and captured their nuclear images with an automated fluorescence microscope. We selected genes that induced greater than 3-fold increase in the percentage of apoptotic nuclei compared with vector-transfected cells. The full length genes C10orf61, MGC 26717, and FLJ13855 were identified as candidate proapoptotic genes, and their apoptotic effects were confirmed by DNA fragmentation ELISAs and Western blotting for caspase-7 and PARP. We conclude that a subcellular image-based apoptotic screen is useful for identifying genes with proapoptotic activity. PMID- 17464194 TI - SM22alpha is required for agonist-induced regulation of contractility: evidence from SM22alpha knockout mice. AB - The present study was undertaken to determine whether SM22alpha participates in the regulation of vascular smooth muscle contractility using SM22alpha knockout mice and, if so, to investigate the mechanisms involved. Aortic ring preparations were mounted and equilibrated in organ baths for 60 min before observing contractile responses to 50 mM KCl, and then exposed to contractile agents such as phenylephrine and phorbol ester. Measurement of isometric contractions using a computerized data acquisition system was combined with molecular or cellular experiments. Interestingly, the aortas from SM22alpha-deficient mice (SM22(-/ LacZ)) displayed an almost three-fold increase in the level of SM22beta protein compared to wild-type mice, but no change in the levels of caldesmon, actin, desmin or calponin. Ca2+-independent contraction in response to phenylephrine or phorbol ester was significantly decreased in the SM22alpha-deficient mice, whereas in the presence of Ca2+ neither contraction nor subcellular translocation of myosin light chain kinase (MLCK) in response to phenylephrine or 50 mM KCl was significantly affected. A decrease in phosphorylation of extracellular signal regulated kinase (ERK) 1/2 was observed in the SM22alpha-deficient mice and this may be related to the decreased vascular contractility. Taken together, this study provides evidence for a pivotal role of SM22alpha in the regulation of Ca2+ independent vascular contractility. PMID- 17464195 TI - Complete mitochondrial genome of a troglobite millipede Antrokoreana gracilipes (Diplopoda, Juliformia, Julida), and juliformian phylogeny. AB - The complete mitochondrial genome of a troglobite millipede Antrokoreana gracilipes (Verhoeff, 1938) (Dipolopoda, Juliformia, Julida) was sequenced and characterized. The genome (14,747 bp) contains 37 genes (2 ribosomal RNA genes, 22 transfer RNA genes and 13 protein-encoding genes) and two large non-coding regions (225 bp and 31 bp), as previously reported for two diplopods, Narceus annularus (order Spirobolida) and Thyropygus sp. (order Spirostreptida). The A + T content of the genome is 62.1% and four tRNAs (tRNA(Ser(AGN)), tRNA(Cys), tRNA(Ile) and tRNA(Met)) have unusual and unstable secondary structures. Whereas Narceus and Thyropygus have identical gene arrangements, the tRNA(Thr) and tRNA(Trp) of Antrokoreana differ from them in their orientations and/or positions. This suggests that the Spirobolida and Spirostreptida are more closely related to each other than to the Dipolopoda. Three scenarios are proposed to account for the unique gene arrangement of Antrokoreana. The data also imply that the Duplication and Nonrandom Loss (DNL) model is applicable to the order Julida. Bayesian inference (BI) and maximum likelihood (ML) analyses using amino acid sequences deduced from the 12 mitochondrial protein-encoding genes (excluding ATP8) support the view that the three juliformian members are monophyletic (BI 100%; ML 100%), that Thyropygus (Spirostreptida) and Narceus (Spirobolida) are clustered together (BI 100%; ML 83%), and that Antrokoreana (Julida) is a sister of the two. However, due to conflict with previous reports using cladistic approaches based on morphological characteristics, further studies are needed to confirm the close relationship between Spirostreptida and Spirobolida. PMID- 17464196 TI - Marker-assisted genotype analysis of bulb colors in segregating populations of onions (Allium cepa). AB - Bulb color in onions (Allium cepa) is an important trait whose complex inheritance mechanism involves epistatic interactions among major color-related loci. Recent studies revealed that inactivation of dihydroflavonol 4-reductase (DFR) in the anthocyanin synthesis pathway was responsible for the color differences between yellow and red onions, and two recessive alleles of the anthocyanidin synthase (ANS) gene were responsible for a pink bulb color. Based on mutations in the recessive alleles of these two genes, PCR-based markers for allelic selection were developed. In this study, genotype analysis of onions from segregating populations was carried out using these PCR-based markers. Segregating populations were derived from the cross between yellow and red onions. Five yellow and thirteen pink bulbs from one segregating breeding line were genotyped for the two genes. Four pink bulbs were heterozygous for the DFR gene, which explains the continuous segregation of yellow and pink colors in this line. Most pink onions were homozygous recessive for the ANS gene, except for two heterozygotes. This finding indicated that the homozygous recessive ANS gene was primarily responsible for the pink color in this line. The two pink onions, heterozygous for the ANS gene, were also heterozygous for the DFR gene, which indicated that the pink color was produced by incomplete dominance of a red color gene over that of yellow. One pink line and six other segregating breeding lines were also analyzed. The genotyping results matched perfectly with phenotypic color segregation. PMID- 17464197 TI - Hydroquinone, a reactive metabolite of benzene, reduces macrophage-mediated immune responses. AB - Hydroquinone is a toxic compound and a major benzene metabolite. We report that it strongly inhibits the activation of macrophages and associated cells. Thus, it suppressed the production of proinflammatory cytokines [tumor necrosis factor (TNF)-alpha, interleukin (IL)-1beta, IL-3, IL-6, IL-10, IL-12p40, IL-23], secretion of toxic molecules [nitric oxide (NO) and reactive oxygen species (ROS)] and the activation and expression of CD29 as judged by cell-cell adhesion and surface staining experiments. The inhibition was due to the induction of heme oxygenase (HO)-1 in LPS-activated macrophages, since blocking HO-1 activity with ZnPP, an HO-1 specific inhibitor, abolished hydroquinone's NO inhibitory activity. In addition, hydroquinone and inhibitors (wortmannin and LY294002) of the phosphatidylinositol-3 kinase (PI3K)/Akt pathway had very similar inhibitory effects on LPS-induced and CD29-mediated macrophage responses, including the phosphorylation of Akt. Therefore, our data suggest that hydroquinone inhibits macrophage-mediated immune responses by modulating intracellular signaling and protective mechanisms. PMID- 17464198 TI - Comprehensive analysis of the expression of twenty-seven beta-1, 3-glucanase genes in rice (Oryza sativa L.). AB - Plant beta-1, 3-glucanases are involved in plant defense and in development. Very little data are available on the expression of rice glucanases both in developmental tissues and under various stresses. In this study, we cloned and characterized twenty-seven rice beta-1, 3-glucanases (OsGlu) from at total of 71 putative glucanases. The OsGlu genes were obtained by PCR from a cDNA library and were classified into seven groups (Group I to VII) according to their DNA or amino acid sequence homology. Analysis of the expression of the twenty-seven OsGlu genes by Northern blotting revealed that they were differentially expressed in different developmental tissues as well as in response to plant hormones, biotic stress, high salt etc. OsGlu11 and 27 in Group IV were clearly expressed only in stem and leaf and were also induced strongly by SA (5 mM), ABA (200 microM), and M. grisea. OsGlu1, 10, 11, and 14 were induced earlier and to higher levels in incompatible M. grisea interaction than in compatible one. Taken together, our findings suggest that the twenty-seven rice OsGlu gene products play diverse roles not only in plant defense but also in hormonal responses and in development. PMID- 17464199 TI - TTF-1 expression in PACAP-expressing retinal ganglion cells. AB - In mammals light input resets the central clock of the suprachiasmatic nucleus by inducing secretion of pituitary adenylate cyclase-activating polypeptide (PACAP) from retinal ganglion cells (RGCs). We previously showed that thyroid transcription factor 1 (TTF-1), a homeodomain-containing transcription factor, specifically regulates PACAP gene expression in the rat hypothalamus. In the present study we examined the expression of TTF-1 in PACAP-synthesizing retinal cells. Fluorescence in situ hybridization (FISH) showed that it is abundantly expressed in RGCs of the superior region of the retina, but in only a small subset of RGCs in the inferior region. Double FISH experiments revealed that TTF 1 is exclusively expressed in PACAP-producing RGCs. These results suggest that TTF-1 plays a regulatory role in PACAP-expressing retinal ganglion cells. PMID- 17464200 TI - Identification of genes suitable for DNA barcoding of morphologically indistinguishable Korean Halichondriidae sponges. AB - The development of suitable genetic markers would be useful for defining species and delineating the species boundaries of morphologically indistinguishable sponges. In this study, genetic variation in the sequences of nuclear rDNA and the mitochondrial cytochrome c oxidase subunit 1 and 3 (CO1 and CO3) regions were compared in morphologically indistinguishable Korean Halichondriidae sponges in order to determine the most suitable species-specific molecular marker region. The maximal congeneric nucleotide divergences of Halichondriidae sponges in CO1 and CO3 are similar to those found among anthozoan cnidarians, but they are 2- to 8-fold lower than those found among genera of other triploblastic metazoans. Ribosomal internal transcribed spacer regions (ITS: ITS1 + ITS2) showed higher congeneric variation (17.28% in ITS1 and 10.29% in ITS2) than those of CO1 and CO3. Use of the guidelines for species thresholds suggested in the recent literature indicates that the mtDNA regions are not appropriate for use as species-specific DNA markers for the Halichondriidae sponges, whereas the rDNA ITS regions are suitable because ITS exhibits a low level of intraspecific variation and a relatively high level of interspecific variation. In addition, to test the reliability of the ITS regions for identifying Halichondriidae sponges by PCR, a species-specific multiplex PCR primer set was developed. PMID- 17464201 TI - A modeling study of co-transcriptional metabolism of hnRNP using FMR1 gene. AB - Since molecular structure of hnRNP is not available in foreseeable future, it is best to construct a working model for hnRNP structure. A geometric problem, assembly of 700 +/- 20 nucleotides with 48 proteins, is visualized by a frame work in which all the proteins participate in primary binding, followed by secondary, tertiary and quaternary binding with neighboring proteins without additional import. Thus, 40S hnRNP contains crown-like secondary structure (48 stem-loops) and appearance of 6 petal (octamers) rose-like architectures. The proteins are wrapped by RNA. Co-transcriptional folding for RNP fibril of FMR1 gene can produce 2,571 stem-loops with frequency of 1 stem-loop/15.3 nucleotides and 53 40S hnRNP beaded structure. By spliceosome driven reactions, there occurs removal of 16 separate lariated RNPs, joining 17 separate beaded exonic structures and anchoring EJC on each exon junction. Skipping exon 12 has 5'GU, 3'AG and very compact folding pattern with frequency of 1 stem-loop per 12 nucleotides in short exon length (63 nucleotides). 5'end of exon 12 contains SS (Splicing Silencer) element of UAGGU. In exons 10, 15 and 17 where both regular and alternative splice sites exist, SS (hnRNP A1 binding site) is observed at the regular splicing site. End products are mature FMR-1 mRNP, 4 species of Pri microRNAs derived from introns 7,9,15 and 3'UTR of exon17, respectively. There may also be some other regulatory RNAs containing ALU/Line elements as well. PMID- 17464202 TI - Differential regulation of the genes of the Streptococcus pneumoniae dnaK operon by Ca++. AB - DnaK is a major antigen in Streptococcus pneumoniae, and is induced by a minor shift in temperature (30 to 37 degrees ) but not by ethanol shock. Although HrcA in the presence of Ca++ represses the expression of both groEL and hrcA, the control of transcription of the dnaK operon is not completely understood. In this study, the dnaK operon of S. pneumoniae (5' hrcA-grpE-dnaK-dnaJ) was cloned and analyzed. It contains large intergenic regions in grpE/dnaK and dnaK/dnaJ. Pulse labeling with [35S]-methionine and immunoblot analyses revealed the presence of higher levels of DnaK than of HrcA even in the presence of Ca++ after heat shock suggesting that Ca++ differentially regulates the heat shock responses of hrcA and dnaK. By blocking de novo mRNA synthesis with rifampin it was shown that neither the hrcA nor the groEL transcripts were stabilized by heat shock even though dnaK transcripts were stabilized. We conclude that S. pneumoniae uses fine regulation of the transcription of the individual genes of the tetracistronic dnaK operon to cope with the various stresses experienced during infections. PMID- 17464203 TI - Microphthalmia-associated transcription factor polymorphisms and association with bone mineral density of the proximal femur in postmenopausal women. AB - Osteoporosis is a common metabolic bone disease characterized by low bone mineral density (BMD) with an increased risk of fracture. Low bone mass results from an imbalance between bone formation by osteoblasts and bone resorption by osteoclasts. Microphthalmia-associated transcription factor (MITF) plays a critical role in osteoclast development and thus is an important candidate gene affecting bone turnover and BMD. In order to investigate the genetic effects of MITF variations on osteoporosis, we directly sequenced the MITF gene in 24 Koreans, and identified fifteen sequence variants. Two polymorphisms (+227719C > T and +228953A > G) were selected based on their allele frequencies, and then genotyped in a larger number of postmenopausal women (n = 560). Areal BMD (g/cm2) of the anterior-posterior lumbar spine and the non-dominant proximal femur was measured by dual-energy X-ray absorptiometry. We found that the MITF + 227719C > T polymorphism was significantly associated with low BMD of the trochanter (p = 0.005-0.006) and total femur (p = 0.02-0.03) (codominant and dominant models), while there was no association with BMD of the lumbar spine. The MITF+228953A > G polymorphism was also associated with low BMD of the femoral shaft (p = 0.05) in the recessive model. Haplotype analysis showed that haplotype 3 of the MITF gene (MITF-ht3) was associated with low BMD of the trochanter (p = 0.03-0.05) and total femur (p = 0.05) (dominant and codominant models). Our results suggest that MITF variants may play a role in the decreased BMD of the proximal femur in postmenopausal women. PMID- 17464204 TI - Nucleotide triphosphates inhibit the degradation of unfolded proteins by HslV peptidase. AB - Escherichia coli HslVU is an ATP-dependent protease consisting of two heat shock proteins, the HslU ATPase and HslV peptidase. In the reconstituted enzyme, HslU stimulates the proteolytic activity of HslV by one to two orders of magnitude, while HslV increases the rate of ATP hydrolysis by HslU several-fold. Here we show that HslV alone can efficiently degrade certain unfolded proteins, such as unfolded lactalbumin and lysozyme prepared by complete reduction of disulfide bonds, but not their native forms. Furthermore, HslV alone cleaved a lactalbumin fragment sandwiched by two thioredoxin molecules, indicating that it can hydrolyze the internal peptide bonds of lactalbumin. Surprisingly, ATP inhibited the degradation of unfolded proteins by HslV. This inhibitory effect of ATP was markedly diminished by substitution of the Arg86 residue located in the apical pore of HslV with Gly, suggesting that interaction of ATP with the Arg residue blocks access of unfolded proteins to the proteolytic chamber of HslV. These results suggest that uncomplexed HslV is inactive under normal conditions, but may can degrade unfolded proteins when the ATP level is low, as it is during carbon starvation. PMID- 17464205 TI - Pattern-recognition receptor signaling initiated from extracellular, membrane, and cytoplasmic space. AB - Invading pathogens are recognized by diverse germline-encoded pattern-recognition receptors (PRRs) which are distributed in three different cellular compartments: extracellular, membrane, and cytoplasmic. In mammals, the major extracellular PRRs such as complements may first encounter the invading pathogens and opsonize them for clearance by phagocytosis which is mediated by membrane-associated phagocytic receptors including complement receptors. The major membrane associated PRRs, Toll-like receptors, recognize diverse pathogens and generate inflammatory signals to coordinate innate immune responses and shape adaptive immune responses. Furthemore, certain membrane-associated PRRs such as Dectin-1 can mediate phagocytosis and also induce inflammatory response. When these more forefront detection systems are avoided by the pathogens, cytoplasmic PRRs may play major roles. Cytoplasmic caspase-recruiting domain (CARD) helicases such as retinoic acid-inducible protein I (RIG-I)melanoma differentiation-associated gene 5 (MDA5), mediate antiviral immunity by inducing the production of type I interferons. Certain members of nucleotide-binding oligomerization domain (NOD) like receptors such as NALP3 present in the cytosol form inflammasomes to induce inflammatory responses upon ligand recognition. Thus, diverse families of PRRs coordinately mediate immune responses against diverse types of pathogens. PMID- 17464206 TI - Multiple actions of dimethylsphingosine in 1321N1 astrocytes. AB - N,N-dimethyl-D-erythro-sphingosine (DMS) is an N-methyl derivative of sphingosine and an inhibitor of protein kinase C (PKC) and sphingosine kinase (SK). In the present study, we examined the effects of DMS on intracellular Ca2+ concentration, pH, and glutamate uptake in human 1321N1 astrocytes. DMS increased intracellular Ca2+ concentration and cytosolic pH in a concentration-dependent manner. Pretreatment of the cells with the Gi/o protein inhibitor PTX and the PLC inhibitor U73122 had no obvious effect. However, removal of extracellular Ca2+ with the Ca2+ chelator EGTA or depletion of intracellular Ca2+ stores with thapsigargin impeded the DMS-induced increase of intracellular Ca2+ concentration. Pretreatment of cells with NH4Cl or monensin reduced the DMS induced Ca2+ increase. However, inhibition of the DMS-induced Ca2+ increase with BAPTA did not influence the DMS-induced pH increase. DMS also inhibited glutamate uptake by the 1321N1 astrocytes in a concentration-dependent manner. It also increased intracellular Ca2+ and pH in PC12 neuronal cells. Our observations on the effects of DMS on 1321N1 astrocytes and PC12 neuronal cells point to a physiological role of DMS in the brain. PMID- 17464207 TI - Human erythropoietin induces lung failure and erythrocytosis in transgenic mice. AB - We have expressed human erythropoietin (EPO) in transgenic mice using a recombinant EPO cDNA combined with a partial TPO construct. The gene was microinjected using standard techniques and five mice were detected as transgenic by PCR and further used as founders. The life span of the transgenic founders was much shorter than that of their normal littermates. Most of the tissues of the transgenic founders contained human EPO transcripts as judged by RT-PCR. Especially high expression levels were seen in the liver and lung. EPO protein levels in serum were examined by ELISA and ranged from 266, 414 mIU/ml. The number of red blood cell, white blood cell and hemoglobin in the hEPO transgenic mice was higher than in normal mice. These results indicate that overexpression of hEPO is deleterious and can provoke lung failure and erythrocytosis. PMID- 17464208 TI - The PKA/CREB pathway is closely involved in VEGF expression in mouse macrophages. AB - Cyclic AMP-responsive element binding protein (CREB) is known to be associated with angiogenesis. In the present study we investigated the possible role of CREB in the expression of vascular endothelial growth factor (VEGF) by mouse macrophages. Over-expression of CREB increased VEGF secretion by cells of the RAW264.7 mouse macrophage cell line. It also increased the promoter activity of a mouse reporter driven by the VEGF promoter, while a dominant negative CREB (DN CREB) abrogated the activity, suggesting that CREB mediates VEGF transcription. Forskolin, an adenylyl cyclase activator, stimulated VEGF transcription, and the PKA inhibitor H89 abolished this effect. IFN-gamma, a potent cytokine, stimulated VEGF expression only in part through the PKA-CREB pathway. These results indicate that PKA phosphorylates CREB and so induces VEGF gene expression. An analysis of mutant promoters revealed that one of the putative CREB responsive elements (CREs), at 399 approximately 388 in the promoter, is critical for CREB-mediated VEGF promoter activity, and the significance of this CRE was confirmed by chromatin immunoprecipitation assays. PMID- 17464209 TI - Opposing effects of ERK and p38 MAP kinases on HeLa cell apoptosis induced by dipyrithione. AB - Dipyrithione (2, 2'-dithiobispyridine-1, 1'-dioxide, PTS2), a pyrithione derivate, is highly bactericidal and fungicidal. In this study we examined its apoptotic effect on HeLa cells. PTS2 induced HeLa cell death in a dose and time dependent manner. ERK1/2 and p38 were markedly activated, but little JNK1/2 activation was detected. Suppression of p38 activation by SB203580 reduced the extent of apoptosis of the HeLa cells and also prevented induction of p21, release of cytochrome c, and cleavage of caspase-3 and PARP. Inhibition of ERK1/2 with PD98059 increased apoptosis, indicating that ERK1/2 activation has an anti apoptotic effect on PTS2-induced HeLa cell apoptosis. PTS2 also inhibited murine sarcoma 180 and hepatoma 22 tumor growth in an animal tumor model. Our findings indicate that PTS2 possesses anti-tumor activity, that caspase-3 and poly (ADP ribose) polymerase (PARP) are involved in PTS2-induced HeLa cell apoptosis and that ERK1/2 and p38 have opposing effects on this apoptosis. PMID- 17464210 TI - Rapid diagnosis of CMT1A duplications and HNPP deletions by multiplex microsatellite PCR. AB - Charcot-Marie-Tooth (CMT) disease and hereditary neuropathy with liability to pressure palsies (HNPP) are frequent forms of genetically heterogeneous peripheral neuropathies. Reciprocal unequal crossover between flanking CMT1A-REPs on chromosome 17p11.2-p12 is a major cause of CMT type 1A (CMT1A) and HNPP. The importance of a sensitive and rapid method for identifying the CMT1A duplication and HNPP deletion is being emphasized. In the present study, we established a molecular diagnostic method for the CMT1A duplication and HNPP deletion based on hexaplex PCR of 6 microsatellite markers (D17S921, D17S9B, D17S9A, D17S918, D17S4A and D17S2230). The method is highly time-, cost- and sample-saving because the six markers are amplified by a single PCR reaction and resolved with a single capillary in 3 h. Several statistical and forensic estimates indicated that most of these markers are likely to be useful for diagnosing the peripheral neuropathies. Reproducibility, as determined by concordance between independent tests, was estimated to be 100%. The likelihood that genotypes of all six markers are homozygous in randomly selected individuals was calculated to be 1.6 x 10(-4) which indicates that the statistical error rate for this diagnosis of HNPP deletion is only 0.016%. PMID- 17464211 TI - Comparative analysis of the developmental competence of three human embryonic stem cell lines in vitro. AB - One of the goals of stem cell technology is to control the differentiation of human embryonic stem cells (hESCs), thereby generating large numbers of specific cell types for many applications including cell replacement therapy. Although individual hESC lines resemble each other in expressing pluripotency markers and telomerase activity, it is not clear whether they are equivalent in their developmental potential in vitro. We compared the developmental competence of three hESC lines (HSF6, Miz-hES4, and Miz-hES6). All three generated the three embryonic germ layers, extraembryonic tissues, and primordial germ cells during embryoid body (EB) formation. However, HSF6 and Miz-hES6 readily formed neuroectoderm, whereas Miz-hES4 differentiated preferentially into mesoderm and endoderm. Upon terminal differentiation, HSF6 and Miz-hES6 produced mainly neuronal cells whereas Miz-hES4 mainly formed mesendodermal derivatives, including endothelial cells, leukocyte progenitors, hepatocytes, and pancreatic cells. Our observations suggest that independently-derived hESCs may differ in their developmental potential. PMID- 17464212 TI - Leukotriene synthesis in response to A23187 is inhibited by methyl- beta cyclodextrin in RBL-2H3 cells. AB - Leukotrienes (LTs) are produced by several biosynthetic enzymes including cytosolic phospholipase A2 (cPLA2), 5-lipoxygenase (5-LO), and 5-lipoxygenase activating protein (FLAP) in the perinuclear area. In the present study, we showed that pretreatment with methyl-beta-cyclodextrin (MbetaCD), a cholesterol depleting agent, dramatically reduced the synthesis of LTs in response to A23187 in mast cells. A23187-induced LT synthesis was inhibited by pretreatment with MbetaCD, and this effect was reversed when cholesterol was added. In an approach to identifying the MbetaCD-sensitive protein(s), we observed that FLAP co localized with flotillin-1, a lipid raft marker protein, in the lipid raft-rich low-density region of sucrose gradients. In addition, electron microscopic analysis revealed that FLAP co-localized with flotillin-1. Together, these results suggest that FLAP is present in cholesterol-rich lipid raft-like domains and that its localization in these domains is critical for LT synthesis. PMID- 17464213 TI - 18S ribosomal DNA sequences provide insight into the phylogeny of patellogastropod limpets (Mollusca: Gastropoda). AB - To investigate the phylogeny of Patellogastropoda, the complete 18S rDNA sequences of nine patellogastropod limpets Cymbula canescens (Gmelin, 1791), Helcion dunkeri (Krauss, 1848), Patella rustica Linnaeus, 1758, Cellana toreuma (Reeve, 1855), Cellana nigrolineata (Reeve, 1854), Nacella magellanica Gmelin, 1791, Nipponacmea concinna (Lischke, 1870), Niveotectura pallida (Gould, 1859), and Lottia dorsuosa Gould, 1859 were determined. These sequences were then analyzed along with the published 18S rDNA sequences of 35 gastropods, one bivalve, and one chiton species. Phylogenetic trees were constructed by maximum parsimony, maximum likelihood, and Bayesian inference. The results of our 18S rDNA sequence analysis strongly support the monophyly of Patellogastropoda and the existence of three subgroups. Of these, two subgroups, the Patelloidea and Acmaeoidea, are closely related, with branching patterns that can be summarized as [(Cymbula + Helcion) + Patella] and [(Nipponacmea + Lottia) + Niveotectura]. The remaining subgroup, Nacelloidea, emerges as basal and paraphyletic, while its genus Cellana is monophyletic. Our analysis also indicates that the Patellogastropoda have a sister relationship with the order Cocculiniformia within the Gastropoda. PMID- 17464214 TI - Identification of QTLs associated with physiological nitrogen use efficiency in rice. AB - Demand for low-input sustainable crop cultivation is increasing to meet the need for environment-friendly agriculture. Consequently, developing genotypes with high nutrient use efficiency is one of the major objectives of crop breeding programs. This study was conducted to identify QTLs for traits associated with physiological nitrogen use efficiency (PNUE). A recombinant inbred population (DT RILs) between Dasanbyeo (a tongil type rice, derived from an indica x japonica cross and similar to indica in its genetic make-up) and TR22183 (a Chinese japonica variety) consisting of 166 F8 lines was developed and used for mapping. A frame map of 1,409 cM containing 113 SSR and 103 STS markers with an average interval of 6.5 cM between adjacent marker loci was constructed using the DT RILs. The RILs were cultivated in ordinary-N (N-P2O5-K2O = 100-80-80 kg/ha) and low-N (N-P2O5-K2O= 50-80-80 kg/ha) (100 kg/ha) conditions. PNUE was positively correlated with the harvest index and grain yield in both conditions. Twenty single QTLs (S-QTLs) and 58 pairs of epistatic loci (E-QTLs) were identified for the nitrogen concentration of grain, nitrogen concentration of straw, nitrogen content of shoot, harvest index, grain yield, straw yield and PNUE in both conditions. The phenotypic variance explained by these S-QTLs and E-QTLs ranged from 11.1 to 44.3% and from 16.0% to 63.6% , respectively. The total phenotypic variance explained by all the QTLs for each trait ranged from 35.8% to 71.3%, showing that the expression of PNUE and related characters depends significantly upon genetic factors. Both S-QTLs and E-QTLs may be useful for marker-assisted selection (MAS) to develop higher PNUE genotypes. PMID- 17464215 TI - Transcriptional activator elements for curtovirus C1 expression reside in the 3' coding region of ORF C1. AB - Beet curly top virus (BCTV) and Beet severe curly top virus (BSCTV), members of curtoviruses, encode seven open reading frames (ORFs) within a approximately 3 kb genome. One of these viral ORFs, C1, is known to play an important role in the early stage of viral infection in plants during initiation of viral DNA replication. We used promoter:: reporter (beta-glucuronidase) gene fusions in transgenic Ara-bidopsis to identify the putative promoter region of BCTV ORF C1. Unlike other geminiviruses, the intergenic region of BCTV was not sufficient to promote C1 expression in transgenic plants. When sequences extending into the coding region of C1 were tested, strong expression of the reporter protein was observed in vascular tissues of transgenic plants. This expression was not dependent on the presence of the intergenic regions or proximal 5' portions of the C1 coding region. Transgenic plants expressing a reporter gene under control of the putative complete C1 promoter were inoculated with virus to determine if any viral transcript affected C1 expression. Virus inoculated plants did not show any altered pattern or change in of reporter gene expression level. These results suggest that (1) important transcriptional activator elements for C1 expression reside in the 3' portion of C1 coding area itself, (2) C1 protein does not auto regulate its own expression and (3) C1 expression of two curtoviruses is controlled differently compared to other geminiviruses. PMID- 17464216 TI - Short-term cold exposure may cause a local decrease of neuropeptide Y in the rat hypothalamus. AB - Neuropeptide Y (NPY) is an orexigenic and hypothermic peptide. To understand its role in hypothermic conditions, male rats were placed in a 24 degrees C or 4 degrees C air chamber for 1.5 h. The expression of c-Fos protein, and NPY mRNA and protein, was analyzed in the hypothalamus 1 h-2 h later. The cold treatment increased the number of c-Fos-immunoreactive cells in the paraventricular hypothalamic nucleus (PVN) and arcuate nucleus (ARC). At the same time it decreased the density of NPY-immunoreactive components in the PVN, dorsomedial hypothalamic nucleus and ARC, as well as of NPY transcripts in the PVN and ARC. No colocalization of c-Fos with NPY was detected. These results suggest that short-term cold exposure should reduce indirectly NPY production in some hypothalamic nuclei to facilitate thermogenesis without inducing feeding behavior. PMID- 17464217 TI - PMA activates Stat3 in the Jak/Stat pathway and induces SOCS5 in rat brain astrocytes. AB - Suppressors of cytokine signaling (SOCS) family members are negative feedback regulators of the Jak/Stat pathway, which is an essential inflammatory signaling pathway. We investigated expression of eight members of the SOCS family in rat astrocytes, using two inflammatory stimulants, PMA and IFN-gamma. Only a few SOCS genes were induced by both stimulants, and we detected an increase in SOCS5 protein with PMA. PMA activated the Jnk, Erk, p38, and Jak/Stat signal pathways. In addition, it increased the level of activated-Stat3 resulting from tyrosine phosphorylation. A gel-shift assay showed that a protein in nuclear extracts from PMA-treated cells was able to bind to Stat binding elements. These results suggest that activated Stat3 binds to SOCS promoters and leads to their transcriptional induction. PMID- 17464218 TI - Modified suppression subtractive hybridization identifies an AP2-containing protein involved in metal responses in Physcomitrella patens. AB - The moss Physcomitrella patens has two life cycles, filamentous protonema and leafy gametophore. A modified from of suppression subtractive hybridization (SSH), mirror orientation selection (MOS), was applied to screen genes differentially expressed in the P. patens protonema. Using reverse Northern blot analysis, differentially expressed clones were identified. The identified genes were involved mainly in metal binding and detoxification. One of these genes was an AP2 (APETALA2) domain-containing protein (PpACP1), which was highly up regulated in the protonema. Alignment with other AP2/EREBPs (Ethylene Responsive Element Binding Proteins) revealed significant sequence homology of the deduced amino acid sequence in the AP2/EREBP DNA binding domain. Northern analysis under various stress conditions showed that PpACP1 was induced by ethephon, cadmium, copper, ABA, IAA, and cold. In addition, it was highly expressed in suspension cultured protonema. We suggest that PpACP1 is involved in responses to metals, and that suspension culture enhance the expression of genes responding to metals. PMID- 17464219 TI - Identification of putative MAPK kinases in Oryza minuta and O. sativa responsive to biotic stresses. AB - The mitogen-activated protein kinase (MAPK) signaling cascade is critical for regulating plant defense systems against various kinds of pathogen and environmental stresses. One component of this cascade, the MAP kinase kinases (MAPKK), has not yet been shown to be induced in plants following biotic attacks, such as those by insects and fungi. We describe here a gene coding for a blast (Magnaporthe grisea)- and insect (Nilaparvata lugens)-responsive putative MAPK kinase, OmMKK1 (Oryza minuta MAPKK 1), which was identified in a library of O. minuta expressed sequence tags (ESTs). Two copies of OmMKK1 are present in the O. minuta genome. They encode a predicted protein with molecular mass 39 kDa and pI of 6.2. Transcript patterns following imbibition of plant hormones such as methyl jasmonic acid (MeJA), ethephone, salicylic acid (SA) and abscisic acid (ABA), as well as exposure to methyl viologen (MV), revealed that the expression of OmMKK1 is related to defense response signaling pathways. A comparative analysis of OmMKK1 and its O. sativa ortholog OsMKK1 showed that both were induced by stress related hormones and biotic stresses, but that the kinetics of their responses differed despite their high amino acid sequence identity (96%). PMID- 17464220 TI - Expression of EuNOD-ARP1 encoding auxin-repressed protein homolog is upregulated by auxin and localized to the fixation zone in root nodules of Elaeagnus umbellata. AB - Root nodule formation is controlled by plant hormones such as auxin. Auxin repressed protein (ARP) genes have been identified in various plant species but their functions are not clear. We have isolated a full-length cDNA clone (EuNOD ARP1) showing high sequence homology to previously identified ARP genes from root nodules of Elaeagnus umbellata. Genomic Southern hybridization showed that there are at least four ARP-related genes in the genome of E. umbellata. The cDNA clone encodes a polypeptide of 120 amino acid residues with no signal peptide or organelle-targeting signals, indicating that it is a cytosolic protein. Its cytosolic location was confirmed using Arabidopsis protoplasts expressing a EuNOD ARP1:smGFP fusion protein. Northern hybridization showed that EuNOD-ARP1 expression was higher in root nodules than in leaves or uninoculated roots. Unlike the ARP genes of strawberry and black locust, which are negatively regulated by exogenous auxin, EuNOD-ARP1 expression is induced by auxin in leaf tissue of E. umbellata. In situ hybridization revealed that EuNOD-ARP1 is mainly expressed in the fixation zone of root nodules. PMID- 17464222 TI - Job redesign and the health care manager. AB - Health care supervisors and managers are often asked to redesign jobs in their departments. Frequently, little information accompanies the directive. This article lists sources of change in work and defines key terms. Also reviewed are factors that supervisors and managers can weigh in their redesigns. The article suggests actions aligned to common problems in the work environment. Finally, guidelines for a practical, step-by-step approach are provided. For health care supervisors and managers, the key to a successful job redesign is to achieve the unique balance of factors that matches the situation. PMID- 17464223 TI - Gossip and nurses: malady or remedy? AB - Gossip is a natural part of every social setting and has a profound influence on organizational behaviors. As the primary care givers in the hospital setting, nurses are the front line in generating and controlling gossip. It is essential that management recognize this dynamic in the nursing workforce so they can be proactive in developing strategies to effectively control gossip. This article highlights the positive and negative aspects of gossip and provides strategies to help nursing professionals effectively manage this workplace issue. Unmanaged gossip can have a negative effect on the workplace by damaging relationships and reputations. Gossip that is managed effectively can have a positive effect on the workplace by building social bonds within the nursing unit. PMID- 17464224 TI - Integration of health care organizations: using the power strategies of horizontal and vertical integration in public and private health systems. AB - Integration in health care attempts to provide all elements in a seamless continuum of care. Pressures influencing development of system-wide integration primarily come from unsustainable cost increases in the United States over the later part of the 20th century and the early 21st century. Promoters of health care integration assume that it will lead to increased effectiveness and quality of care while concurrently increasing cost-effectiveness and possibly facilitating cost savings. The primary focus of this literature review is on the Power Strategies of Horizontal and Vertical Integration. The material presented suggests that vertical integration is most effective in markets where the partners involved are larger and dominant in the regions they serve. The research has also found that integrating health care networks had little or no significant effect on improving overall organizational efficiencies or profits. Capital investment in information technologies still is cost prohibitive and outweighs its benefits to integration efficiencies in the private sector; however, there are some indications of improvements in publicly provided health care. Further research is needed to understand the reasons the public sector has had greater success in improving effectiveness and efficiency through integration than the private sector. PMID- 17464225 TI - Pay for performance: improving quality care. AB - Pay-for-Performance (PFP) is becoming increasingly viewed as a viable means of responding to the ever increasing pressure for economy, continuity, and consistency in the delivery of health care. Pay-for-Performance is an important development in the move toward a more quality-driven health care system. The quality improvement mission of today's PFP programs is to provide realistic incentives encouraging physicians and hospitals to deliver quality health care in an efficient and cost-effective manner. Central to PFP is the enhancement of the essential patient-physician relationship. Existing guidelines developed by the American Medical Association, as well as guidelines published by the Joint Commission on Accreditation of Healthcare Organizations, describe how PFP programs work and how the various players in the health care system--physicians, patients, insurers, payers--are potentially affected by PFP. Several health plans have already adopted functional PFP programs. Overall, PFP programs show promise for improving the quality of care while helping to contain costs. PMID- 17464226 TI - We don't want to go there. AB - Managers must have many skills, among which is the ability to carefully select which challenges and issues should be responded to and which should be better left alone. A wise manager listens to the troubles of staff and concerns in the workplace and then selects which problems to respond to and when. Sometimes, "We don't want to go there" is the appropriate reaction and limits further conversation and energy on the topic. Because the statement cuts others off, it is important to use this phrase with deliberation and tact. PMID- 17464227 TI - Discharging patients earlier in the day: a concept worth evaluating. AB - Patient discharges from the hospital often occur late in the day and are frequently clustered after 4 PM. When inpatients leave earlier in the day, quality is improved because new admissions awaiting beds are able to leave the emergency department sooner and emergency department waiting room backlog is reduced. Nursing staff, whose work patterns traditionally result in high activity of discharge and admission between 5 PM and 8 PM, benefit by spreading out their work across a longer part of the day. Discharging patients earlier in the day also has the potential to increase patient satisfaction. Despite multiple stakeholders in the discharge planning process, physicians play the most important role. Getting physician buy-in requires an ability to teach physicians about the concept of early-in-the-day discharges and their impact on the process. We defined a new physician-centered discharge planning process and introduced it to an internal medicine team with an identical control team as a comparison. Discharge time of day was analyzed for 1 month. Mean time of day of discharge was 13:39 for the intervention group versus 15:45 for the control group (P<.001). If reproduced successfully, this process could improve quality at an important transition point in patient care. PMID- 17464228 TI - Nurse-nurse manager conflict: how do nurse managers manage it? AB - This study was conducted to determine the conflict management styles used by nurse managers when in conflict with nurses and to ascertain the differences between sociodemographic characteristics and the style used. The study was conducted on 71 nurse managers working in 3 different hospitals in Antalya (Turkey). A personal information form and Rahim Organizational Conflict Inventory II were used for data collection. The most commonly used style by nurse managers when in conflict was integrating, and the least commonly used was avoiding. There was no difference between the style used and the nurses' educational status, marital status, and years of employment. The obliging style was used the most by those 45 years and older and those with 20 years or more of management experience, whereas the avoiding style was used by nurse managers with 6 to 10 years of management experience. This study suggests that nurse managers who are older and have management experience need to be encouraged to use effective conflict management and that there is a need for further research of the subject of factors that affect conflict management style. PMID- 17464230 TI - Creating a culture for health care quality and safety. AB - Approximately 67% of hospital quality indicators require some type of laboratory testing to monitor compliance. Unfortunately, in many hospitals, laboratory data information systems remain an untapped resource in eliminating medical errors and improving patient safety. Using case scenarios, this article demonstrates potential consequences for patient safety and quality of care when information sharing between medical technologists and nurses is not a part of a hospital's culture. The outcome for this patient could have been avoided if a more inclusive health care quality and safety culture existed. Creating a culture for health care quality and safety requires consensus building by clinical and administrative leaders. Consensus building occurs by managing relationships among and between a team of independent, autonomous physicians, nurses, allied health professionals, and health care administrators. These relationships are built on mutual respect and effective communication. Creating a quality culture is a challenging but necessary prerequisite for eliminating medical errors and ensuring patient safety. Physician leaders promoting and advancing cultural change in clinical care from one of exclusive decision making authority to a culture that is based on shared decision making are a necessary first step. Shared decision making requires mutual respect, trust, confidentiality, responsiveness, empathy, effective listening, and communication among all clinical team members. Physician and administrative leaders with a focus on patient safety and a willingness to change will ensure a culture of health care quality and safety. PMID- 17464231 TI - Does having a "brand" help you lead others? AB - Every manager has the opportunity to develop a personal brand of unique characteristics that are valuable in his or her own right. Recognizing a personal brand and developing it to its fullest can enhance leadership potential. These qualities enable others to notice and follow the leader and can enhance cooperation. Credibility is key to developing a personal brand. Then come style, consistency, and change. A brand can enable the manager to connect with others. Use of technology can enhance the use of a brand because cyberspace promotes communication. Other necessities are using organization, selling vision, sharing information, and staying personal. PMID- 17464232 TI - Conducting the employee selection interview: how to do it effectively while avoiding legal obstacles. AB - Employment interviewing as it is known today is an essential process but one fraught with potential traps and legal pitfalls. On balance, it is a far-from perfect means of selecting employees, but it remains the best such means available. Effective interviewing requires thorough preparation, including knowledge of how to seek out the most helpful kinds of information available; complete information about the position as it presently exists; and detailed knowledge of what kinds of questions can or cannot legally be asked. Effective interviewing also depends on the development of one's ability to seek out intangible information as well as factual information and use all that is learned, recognizing that the well-cultivated "gut feel" is fully as important as "facts" in evaluating an employment applicant. PMID- 17464233 TI - A manager asks: what to do with the troublesome employee? PMID- 17464234 TI - How does open-mouth breathing influence upper airway anatomy? AB - OBJECTIVES/HYPOTHESIS: Open-mouth breathing during sleep may increase the severity of obstructive sleep apnea (OSA) and complicate nasal continuous positive airway pressure (CPAP) therapy in patients with OSA. The aim of this study was to assess the effect of open-mouth breathing on upper airway anatomy using lateral cephalometry and fiberoptic nasopharyngoscopy. STUDY DESIGN: This was a cross-sectional study. METHODS: Lateral cephalometry and fiberoptic nasopharyngoscopy were carried out on 28 subjects with a mean age of 36.7 years. We compared the effect of the mouth being open or closed on the results in lateral cephalometry (pharyngeal length, distance between the mandible and hyoid bone [MP-H], angles from the sella-nasion to mandibular points A and B [SNA, SNB, respectively], retropalatal, retroglossal, and hypopharyngeal distance) and fiberoptic nasopharyngoscopy (retropalatal and retroglossal cross-sectional area). RESULTS: On lateral cephalometric measurements, retropalatal distance (P = .000), retroglossal distance (P = .000), and MP-H (P = .002) were lower with mouth open, and pharyngeal length (P = .000) was greater. However, there were no significant differences in SNA and hypopharyngeal distance. On fiberoptic nasopharyngoscopy, retropalatal (P = .005) and retroglossal (P = .000) cross sectional areas were significantly reduced with the mouth open. CONCLUSIONS: Open mouth breathing is associated with reduction of the retropalatal and retroglossal areas, lengthening of the pharynx and shortening of the MP-H in the upper airway. We suggest that knowledge of these anatomic changes improves our understanding of the increase of OSA severity and the low adherence to nasal CPAP therapy in mouth breathers. PMID- 17464235 TI - Evolution of mild obstructive sleep apnea after different treatments. AB - STUDY OBJECTIVES: To evaluate the prognosis of mild obstructive sleep apnea in relation to different treatment modalities. STUDY DESIGN: An open, retrospective, longitudinal follow-up study. METHODS: Fifty adult patients diagnosed and treated for mild obstructive sleep apnea at the Department of Otorhinolaryngology at Kuopio University Hospital between 1998 and 2004 had a control polysomnography in 2005. The changes in apnea-hypopnea index (AHI) were observed in untreated (n = 28), operative (n = 11), and continuous positive airway pressure (n = 11) treatment groups at a long-term follow-up visit. RESULTS: The mean follow-up period was 4 (range, 1.3-9.0; SD, 1.9) years. The untreated patients had a statistically significant increase in AHI (13.3, SD 18.3) at the follow-up. Half of these patients developed a moderate or severe degree of sleep apnea, and only 11% were cured. In patients who were treated with continuous positive airway pressure, the degree of obstructive sleep apnea became worse in 64% of cases, and in 27% of patients, the AHI returned to normal (<5). The degree of obstructive sleep apnea in operated patients deteriorated only in 18%, and in 27% of the patients, the AHI returned to normal (<5). CONCLUSIONS: Mild obstructive sleep apnea has a natural tendency to worsen with time. Active treatment of mild obstructive sleep apnea appears, therefore, to be advisable. PMID- 17464236 TI - Management of facial nerve schwannoma in patients with favorable facial function. AB - OBJECTIVE: We report six cases of facial nerve schwannomas in which surgical management allowed the preservation of facial nerve function. Specifically, this paper reports that a stripping surgery may provide favorable functional outcomes. STUDY DESIGN: A retrospective review of preoperative and postoperative data for six patients with facial nerve schwannoma that had normal facial nerve function or a House-Brackmann grade II facial palsy before the surgery. METHODS: Stripping surgery, which removed the schwannoma from the remaining nerve fascicle, was attempted on the six patients. Postoperative facial nerve function and imaging (magnetic resonance imaging) were evaluated. RESULTS: Stripping surgery with gross total tumor removal of the mass was performed in four cases. In the two remaining cases, the stripping surgery was not possible, and decompression alone was performed. Favorable preservation of facial function was achieved in all six cases. CONCLUSION: It was possible to preserve facial function after surgery to remove facial nerve schwannoma. We suggest that stripping surgery, focused on the preservation of continuity of the facial nerve, may be attempted for facial nerve schwannoma in which favorable facial function has been preserved. PMID- 17464237 TI - Possible role of equol status in the effects of isoflavone on bone and fat mass in postmenopausal Japanese women: a double-blind, randomized, controlled trial. AB - OBJECTIVE: Equol is more biologically active than its precursor daidzein, which is the principal isoflavone found in soybean. There are interindividual differences in the ability to produce equol; these may lead to differences in the effects of isoflavone intervention on human health. This study aimed to investigate whether the effects of soy isoflavones on bone and fat mass are related to an individual's equol status. DESIGN: We performed a 1-year double blind, randomized trial to compare the effects of isoflavone (75 mg of isoflavone conjugates/day) with those of placebo on bone mineral density, fat mass, and serum isoflavone concentrations in early postmenopausal Japanese women who were classified based on their equol-producer phenotype. RESULTS: After 1 year, the isoflavone intervention significantly increased the serum equol concentration in the equol producers but not in the nonproducers. In the isoflavone group, the annualized changes in the bone mineral density of the total hip and intertrochanteric regions were -0.46% and -0.04%, respectively, in the equol producers and -2.28% and -2.61%, respectively, in the nonproducers; these values were significantly different (P<0.05 for both the regions). Significant differences were observed between the equol producers and nonproducers in the isoflavone group with regard to the annualized changes in the fat mass. No significant difference in the annualized changes in bone mineral density and fat mass was observed between the equol producers and nonproducers in the placebo group. CONCLUSIONS: Our data suggest that the preventive effects of isoflavones on bone loss and fat accumulation in early postmenopausal women depend on an individual's equol-producing capacity. PMID- 17464240 TI - Cancer as a manifestation of aberrant chromatin structure. AB - In this article we review many important epigenetic changes in early carcinogenesis and discuss the possibility of these alterations being targeted for therapeutic intervention in the future. Both regional DNA methylation and global chromatin packaging are interrelated partners that function in concert to control gene transcription. We first summarize briefly DNA methylation and its role in gene expression. Then, we focus on how the DNA is packaged into chromatin and the tight relationship between chromatin and DNA methylation. A more complete understanding of these key, regulatory events is vital in approaching a more rational drug therapy to various malignancies. PMID- 17464241 TI - Cancer susceptibility: epigenetic manifestation of environmental exposures. AB - Cancer is a disease that results from both genetic and epigenetic changes. Discordant phenotypes and varying incidences of complex diseases such as cancer in monozygotic twins as well as genetically identical laboratory animals have long been attributed to differences in environmental exposures. Accumulating evidence indicates, however, that disparities in gene expression resulting from variable modifications in DNA methylation and chromatin structure in response to the environment also play a role in differential susceptibility to disease. Despite a growing consensus on the importance of epigenetics in the etiology of chronic human diseases, the genes most prone to epigenetic dysregulation are incompletely defined. Moreover, neither the environmental agents most strongly affecting the epigenome nor the critical windows of vulnerability to environmentally induced epigenetic alterations are adequately characterized. These major deficits in knowledge markedly impair our ability to understand fully the etiology of cancer and the importance of the epigenome in diagnosing and preventing this devastating disease. PMID- 17464242 TI - DNA methyltransferase inhibitors for cancer therapy. AB - Aberrant DNA methylation patterns, including hypermethylation of tumor suppressor genes, have been described in many human cancers. These epigenetic mutations can be reversed by DNA methyltransferase inhibitors, which provide novel opportunities for cancer therapy. Clinical concepts for epigenetic therapies are currently being developed by using azanucleosides for the treatment of leukemias and other tumors. These trials will greatly benefit from the inclusion of molecular markers for monitoring epigenetic changes in patients and for maximizing biologic responses. In addition, novel inhibitors need to be developed that result in a direct and specific inhibition of DNA methyltransferase activity. Several recent developments indicate that rational design of small molecule DNA methyltransferase inhibitors is feasible and that this approach can result in the establishment of novel drug candidates. The use of novel DNA methyltransferase inhibitors in clinical trials that allow monitoring of drug induced DNA methylation changes should provide the foundation for improved epigenetic cancer therapies. PMID- 17464243 TI - Histone deacetylase inhibitors: biology and mechanism of action. AB - Histone deacetylases (HDACs) and histone acetyltransferases are enzymes that regulate chromatin structure and function through the removal and addition, respectively, of the acetyl group from the lysine residues of core nucleosomal histones. This posttranslational modification of histones is an important process in the regulation of gene expression. Aberrant expression and recruitment and disrupted activities of HDACs and histone acetyltransferases have been found in malignant tissues, implicating their involvement in cancer. HDAC inhibitors (HDACIs) function through diverse mechanisms, including the promotion of cell cycle arrest and apoptosis and the inhibition of angiogenesis. Malignant cells appear more sensitive to the proapoptotic effects of HDACIs, underscoring the therapeutic potential of these agents. Multiple HDACIs are currently under investigation in clinical trials, including vorinostat (suberoylanilide hydroxamic acid), which was recently approved by the U.S. Food and Drug Administration for the treatment of cutaneous manifestations of cutaneous T-cell lymphoma in patients with progressive, persistent, or recurrent disease on or after 2 systemic therapies. PMID- 17464244 TI - Histone deacetylase inhibitors and demethylating agents: clinical development of histone deacetylase inhibitors for cancer therapy. AB - The histone deacetylase inhibitors are a new class of agents that are currently in various stages of clinical development. Clinical trials have demonstrated activity, urging further investigation. At the same time, it has been discovered that these agents have their own challenges. In this review, we discuss clinical data gathered to date, combination therapies designed to increase efficacy, and toxicities attributed to this new class of agents. PMID- 17464245 TI - DNA demethylating agents and histone deacetylase inhibitors in hematologic malignancies. AB - The pivotal role of aberrant promoter methylation in gene silencing and cancer development has fueled the interest in DNA methyltransferase inhibitors as novel anticancer drugs. Modulation of gene expression through targeting of epigenetic marks is one of the emerging and promising strategies that has demonstrated successful clinical outcome in hematologic malignancies. Epigenetic modifiers, including DNA methyltransferase inhibitors and histone deacetylase inhibitors, have demonstrated significant clinical activity; several are or are likely to soon be approved by the U.S. Food and Drug Administration. However, the exact mechanism of the clinical response achieved is not fully understood. This review focuses on the pharmacology of the known DNA methyltransferase and histone deacetylase inhibitors and their potential as promising anticancer drugs. PMID- 17464246 TI - Targeting aberrant chromatin structure in colorectal carcinomas. AB - Epigenetic processes such as DNA methylation and histone modifications are now recognized as critical events for regulation of gene expression in mammalian cells and affect gene function without a change in coding sequence. Neoplastic cells often show profound epigenetic alterations that contribute to tumorigenesis by altering expression of critical genes. In colorectal tumorigenesis, detailed analysis led to a hypothesis on a critical role for epigenetic changes in age related cancer susceptibility and separately identified a distinct phenotype termed the CpG island methylator phenotype. CpG island methylator phenotype positive colorectal cancers have significant associations with female sex, older age, proximal location, mucinous histology, KRAS and BRAF mutations, wild-type p53, and microsatellite instability. Histone modifications that affect chromatin structures are also closely implicated in tumor suppressor gene inactivation and DNA methylation and histone modifications seem to form reinforcing networks for stable gene silencing. Much of the excitement in this field relates to the possibility of therapeutic reversal of epigenetic changes by chromatin-modifying drugs. In CpG island methylator phenotype-positive colorectal cancers, DNA methylation inhibitors restore key silenced pathways in vivo (eg, mismatch repair defects), and hypomethylation can largely abolish tumorigenesis in a mouse model. Drugs that inhibit DNA methylation and histone deacetylation are in use in the clinic and should be tested in colorectal malignancy. PMID- 17464247 TI - Utilization of chromatin remodeling agents for lung cancer therapy. AB - Lung cancer is a disease with enormous global medical and economic impact that remains refractory to conventional treatment modalities. Recent insights regarding mechanisms pertaining to epigenetic regulation of gene expression during malignant transformation, together with the identification of agents that modulate chromatin structure provide new opportunities for the treatment and prevention of this lethal disease. PMID- 17464248 TI - Histone deacetylase inhibitor and demethylating agent chromatin compaction and the radiation response by cancer cells. AB - It now appears that epigenetics plays a central role in transformation, both in vitro and in vivo. The expression and regulation of DNA methylation and the subsequent chromatin structure are significantly altered in tumor cells, suggesting a direct role in the process of in vivo cellular transformation. If epigenetics and posttranslational modifications of histones play a role in transformation, then it seems logical that the genes regulating chromatin compaction may also be molecular targets and markers in profiling tumor cell resistance. Local remodeling of chromatin is a key step in the regulation of gene expression, and altering the expression of these genes might also favorably alter how tumor cells respond to anticancer agents. Several new agents that alter chromatin compaction, either methyltransferase or histone deacetylases inhibitors, are progressing through clinical trials and have shown promising preclinical interactions when combined with radiation. In this review, we discuss the potential for histone deacetylases inhibitors as radiosensitizing agents. PMID- 17464249 TI - An epigenetic approach to cancer etiology. AB - The practice of clinical oncology historically has been rooted in therapy for established cancers, and over the past decade, early detection of the malignancy has occurred increasingly, allowing an increasing chance of cure by surgical intervention. Cancer prevention has been targeted largely to generic reduction of exposure to environmental carcinogens, such as smoking reduction. However, targeted identification of patients at increased risk and therapeutic tailored intervention in those patients have not taken hold in oncology, despite the enormous success of that approach in preventive cardiology. A paradigm of such a strategy for oncology may be the identification of patients with epigenetic alterations in progenitor cells and intervention before the development of the earliest identifiable neoplasms. We review studies of loss of imprinting of insulin-like growth factor 2 in colorectal cancer as an example of such a target for preventive oncology. PMID- 17464250 TI - [Emergency imaging and CT]. PMID- 17464251 TI - [New possibilities to study biliary tree and gallbladder: functional magnetic resonance cholangiography contrast-enhanced with mangafodipir trisodium (Mn DPDP)]. AB - Mangafodipir trisodium (Teslascan) is a hepatobiliary contrast agent that provides noninvasive opacification of the bile ducts. Using this contrast medium combined with a T1-weighted gradient echo enhanced sequence provides functional imaging of the bile ducts. Second-intention MRI was obtained after the usual morphological study of the bile ducts using heavily T2-weighted sequences (SS-FSE Te eff long and SS FSE Te eff short). This method can detect many biliary duct anomalies: biliary leakage in the postoperative context, mapping of bile ducts and the gallbladder in the search for anatomical variants, analysis of biliodigestive or biliobiliary anastomoses, or a dynamic study of bile secretion and excretion. Opacification of the bile ducts has only been possible until now with invasive tests aggravated by a certain co-morbidity rate and their functional study using biliary scintigraphy limited by mediocre spatial resolution. This new possibility provides access not only to morphological imaging, but also to functional imaging with excellent spatial resolution. PMID- 17464252 TI - [Ankle and foot sprains: conventional radiography aspects]. AB - Emergency departments frequently encounter pathology resulting from injury to the foot and ankle, with approximately 6000 case per day in France. In an ankle sprain, 85% of the lesions involve the lateral collateral ligament. Many other, much rarer, types of lesion with different therapeutic consequences can present, however. Interpretation of the initial conventional radiographs is vital to establishing the type lesion and to proposing adapted and rapid treatment. The objective of this article is to review the various osteoarticular and ligament injuries encountered in the foot and the ankle. PMID- 17464253 TI - [CT anatomy of the normal spleen: variants and pitfalls]. AB - Normally, the spleen is located in the left hypochondrium. It is attached by the gastrosplenic and splenorenal ligaments. Its tissue composition made up of red pulp and white pulp explains the heterogenous aspect when contrast medium is injected in the arterial phase. This can make it difficult to detect intrasplenic masses. The adult spleen has no lobulations or incisures; however, the persistence of fissures separating the fetal spleen's lobules can indicate splenic lacerations in a trauma context. The ectopic spleen is a migration of the spleen from its normal anatomic location because its ligaments have not developed properly. The spleen can migrate anywhere in the abdomen or pelvis. The accessory spleen can be found in 10% of the population; it is usually located near the hilum of the main spleen or the pacreatic tail. It can be located in many other places and be confused with a mass. Polysplenia is a complex congenital syndrome associating visceral heterotaxis and concomitant bilateral left-sidedness. The spleen is divided into several splenules of the same size. Splenosis is defined by the implantation of splenic tissues in the peritoneum following traumatic rupture of the spleen. PMID- 17464254 TI - [Impact of computed tomography on patient's care in nontraumatic acute abdomen: 90 patients]. AB - OBJECTIVES: To determine the contribution of computerized tomography (CT) to the management of nontraumatic acute abdomen, to evaluate interobserver agreement and the contribution of CT to cost control, to look for the predictive factors of CT. PATIENTS: and method. Ninety prospectively included patients, admitted for nontraumatic acute abdomen and examined by a surgeon, received CT examination. Diagnosis and treatment 1) envisioned before and 2) defined after CT, and 3) finally retained were compared, and the interobserver agreement was calculated after the second reading. The predictive value of the clinical and biological criteria as well as the radiological criteria characterizing these patients was sought. RESULTS: CT was contributive in 68.9% of cases, with a reliable diagnosis and treatment strategy, defined after CT examination, for 92.2% and 90%, respectively. Interobserver agreement was 93.3%. CT contributed to reducing costs in 15.5% of patients, for an additional cost estimated at 104-139 euros. The positive predictive factors of the CT contribution were age over 70 years, localized symptoms, fever, and high CRP. CONCLUSION: In agreement with the literature, in our study CT appears to be a choice examination to guide patient care in nontraumatic acute abdomen. PMID- 17464255 TI - [Indications for brain scan in patients presenting benign cranial injury]. AB - OBJECTIVE: Validate the clinical criteria, which, when absent, would make it safe to bypass CT scan examination in mild cranial injuries. MATERIAL: and methods. Prospective study including 285 patients with mild cranial injury with a Glasgow score of 15, a normal clinical examination but transitory loss of consciousness or suspected transitory loss of consciousness. The following clinical parameters were systematically reviewed: history of stroke; post-injury headache; post injury vomiting; alcohol, medication, or drug intoxication; clinical signs of cervico-cranio-facial injury; post-injury convulsions; or coagulation impairment. Systematic CT exploration looked for cranial, encephalic, and facial lesions and individualized the lesions requiring neurosurgical or maxillofacial treatment. RESULTS: Of the patients studied, 7% presented a cranioencephalic lesion and 7% a facial bone lesion. Neurosurgical intervention was necessary in 0.4% of the patients and maxillofacial surgery in 2.5%. Patients with a positive CT all had at least one clinical risk factor and patients with cranioencephalic lesions had at least two risk factors present. Had patients with no risk factors not been scanned, 15% of the patients would not have had the CT procedure. CONCLUSION: Selecting CT indications in cases of mild cranial injury with loss of consciousness using a simple and validated evaluation can save 15% of CT procedures without missing any cranial, encephalic, or facial lesions. PMID- 17464256 TI - [Computer-aided detection of lung nodules on thin collimation MDCT: impact on radiologists' performance]. AB - OBJECTIVES: Evaluate the improvement in detecting lung nodules when using multidetector CT (MDCT) computer-assisted diagnosis (CAD). MATERIAL AND METHODS: Three radiologists (R1, R2, R3) with different levels of experience independently interpreted 30 MDCT examinations of the thorax taken for screening purposes, first without and then with CAD. The diagnosis was established by two of the three radiologists interpreting the images together, assisted by the CAD. RESULTS: The consensus reading identified 133 nodules, 61 (46%) of which were 4 mm or larger. The sensitivity values in the detection of nodules before and after using the CAD were 54% and 80% (R1), 38% and 71% (R2), and 70% and 88% (R3), respectively. When considering only the nodules that were 4 mm or larger, the sensitivity values varied before and after using the CAD, from 62% to 95% (R1), from 41% to 84% (R2), and from 74% to 92% (R3). By combining two by two the three radiologists' results obtained without the CAD, the sensitivity values were 65%, 83%, and 77%, respectively, for all the nodules, and 70%, 85%, and 77% for the nodules that were 4 mm or larger. The CAD induced a total of 105 false-positive results, with a mean of 3.5 per examination. CONCLUSION: The lung nodules missed by the radiologist can be detected if the CAD is used as a second reader. The CAD can be at least as beneficial as the use of a second independent reader. PMID- 17464257 TI - [Usefulness of magnetic resonance imaging in Paget disease of the breast]. AB - OBJECTIVE: Evaluate the usefulness of MRI in local staging of Paget disease of the breast. MATERIALS AND METHODS: Clinical, radiological (mammographies, MRI), and histological data were analyzed in six female patients presenting Paget disease of the breast. The criteria for analysis in MRI were the following: morphology of the aureola-nipple plaque (thickening, regularity of the contour) and the type of enhancement after injection of contrast medium (signal intensity/time curve), detection of abnormal enhancing in the mammary gland, and ganglion areas. RESULTS: None of the patients presented a palpable mass or a suspicious anomaly on mammography. On MRI, the aureola-nipple plaque was morphologically abnormal in four cases, with suspicious enhancement in two cases (two cases of ductal carcinoma in situ) and normal enhancement in two cases (one case of ductal carcinoma in situ). In the two other cases, the aureola-nipple plaque was normal (one case of ductal carcinoma in situ). Distant abnormal enhancement of the aureola-nipple plaque was noted in two cases (one case of ductal carcinoma in situ and one benign lesion). CONCLUSION: The MRI aspect of the aureola-nipple plaque in Paget disease shows little concordance with the histological results. MRI can be useful in detecting distant lesions when there is no clinical sign nor a suspicious mammography. PMID- 17464258 TI - [MRI aspect of rare rectal pseudotumor associated with dyschezia: colitis cystica profunda]. PMID- 17464259 TI - [Failure to characterize a suprarenal metastasis of hepatocarcinoma using the percentage of contrast medium washout with CT: a case study]. PMID- 17464260 TI - [Coralliform atheroma of the abdominal aorta: diagnosis and management]. PMID- 17464261 TI - [Tumoral calcinosis: two pediatric case studies]. PMID- 17464262 TI - [Interventional MRI: risk of burns to the patient and radiologist]. PMID- 17464263 TI - [Neonatal MRI and neurodevelopmental outcomes in preterm infants]. PMID- 17464264 TI - [What is your diagnosis? Thoracic extramedullary hematopoiesis in a patient with beta-thalassemia]. PMID- 17464266 TI - Two-dimensional vs three-dimensional imaging in whole body oncologic PET/CT: a Discovery-STE phantom and patient study. AB - AIM: To evaluate the performance of the positron emission tomography (PET)/computed tomography (CT) Discovery-STE (D-STE) scanner for lesion detectability in two-dimensional (2D) and three-dimensional (3D) acquisition. METHODS: A NEMA 2001 Image-Quality phantom with 11 lesions (7-37 mm in diameter) filled with a solution of 18F (lesion/background concentration ratio: 4.4) was studied. 2D and 3D PET scans were sequentially acquired (10 min each) in list mode (LM). Each scan was unlisted into 4, 3 and 2-min scans. Ten [18F]FDG PET oncological patient studies were also evaluated. Each patient underwent a 3D PET/CT whole body scan, followed by a 2D PET scan (4 min LM) and a 3D PET scan (4 min LM) over a single field of view. Both 2D and 3D scans were unlisted in 3 and 2-min scans. Data were evaluated quantitatively by calculating quality measurements and qualitatively by two physicians who judged lesion detectability compared to statistical variations in background activity. RESULTS: Quantitative and qualitative evaluations showed the superiority of 3D over 2D across all measures of quality. In particular, lesion detectability was better in 3D than in 2D at equal scan times and 3D acquisition provided images comparable in quality to 2D in approximately half the time. Interobserver variability was lower in evaluation of 3D scans and lesion shape and volume were better depicted. CONCLUSION: In oncological applications, the D-STE system demonstrated good performance in 2D and 3D acquisition, while 3D exhibited better image quality, data accuracy and consistency of lesion detectability, resulting in shorter scan times and higher patient throughput. PMID- 17464267 TI - Somatostatin receptor PET imaging with Gallium-68 labeled peptides. AB - Imaging somatostatin receptor status with 68Ga labeled peptides has progressed rapidly over the last several years. It has generated great interest, and stimulated further research into the development of DOTA-derivative peptides. It has expanded our knowledge of receptor imaging and enhanced our appreciation of the difference between receptor-based and metabolic imaging, as well as more in depth evaluation of tumor biology. The availability of the 68Ge/68Ga generator provides an attractive alternative to cyclotron-based positron-emitters, especially if kit-based radiopharmaceutical formulations based upon 68Ga are developed in the future. PMID- 17464268 TI - PET in the diagnostic evaluation of adrenal tumors. AB - Early experience with positron emission tomography (PET) has provided exciting results in the evaluation of a broad spectrum of neoplasms, to include primary adrenal tumors, their metastases and metastatic disease to the adrenal glands. By virtue of the well-recognized propensity of malignancies to preferentially use glycolysis as an important energy source and the stimulation of mechanisms designed to absorb substrate glucose, the glucose analog, [18F]fluorodeoxyglucose, has become a successful radiopharmaceutical in the scintigraphic evaluation of adrenal tumors. Building upon prior experience gained with imaging the adrenal gland, other positron-labeled radiopharmaceuticals are finding their way into clinical use. The 11b-hydroxylase inhibitor, metomidate labeled with 11C has been used to scintigraphically identify tissues of adrenocortical origin, to accurately identify recurrent and metastatic adrenocortical carcinoma and may be useful in assessing the malignant potential of these tumors and predicting survival in afflicted patients. Adrenomedulla imaging with 11C- and 18F-labeled catecholamines and catecholamine analogs draws heavily from the experience gained from predecessor compounds, labeled with single photon emitting isotopes and, in some instances, single photon emission tomography, and has been shown to depict with high efficacy pheochromocytomas, neuroblastomas and other neoplasms of neural crest origin. Additional structural and functional information provided by computed tomography (CT), performed as part of hybrid PET/CT imaging directly complements PET and adds measurable diagnostic value in the evaluation of adrenal tumors. PMID- 17464269 TI - PET in genitourinary tract cancers. AB - Genitourinary (GU) tract cancers comprise a variety of tumors, which includes some of the most common malignancies in men and women. As a result of the importance of GU neoplasms and the success of positron emission tomography (PET) in imaging and staging cancer, PET with fluorodeoxyglucose (FDG) has been used to depict and stage ovarian, cervical and testicular cancers. The early success of FDG PET in imaging GU tumors is tempered by the fact that some neoplasms of GU origin do not accumulate sufficient FDG for successful imaging. As a result, alternative agents, such as [11C]choline and [11C]acetate, have been used to image prostate cancer and may have utility in bladder cancer, while other PET agents are currently under active evaluation for this and other GU neoplasms. In this paper, we review the current literature and our experience in role of PET in imaging cancers of the GU tract. PMID- 17464270 TI - Pitfalls in PET/CT interpretation. AB - Positron emission tomography (PET) has become an integral part of the management of patients with cancer as well as some cardiac and neurological diseases. 18F fluorodeoxyglucose (FDG) PET is commonly used to stage cancer patients after initial diagnosis, but is increasingly used at other points in the patient's management, including the assessment of treatment response and detecting recurrent disease. In common with most imaging techniques there are pitfalls in interpretation of PET and PET/computed tomography (CT) studies. Many potential pitfalls and artefacts have previously been described with 18F-FDG PET imaging, but more continue to become apparent as worldwide experience increases. In addition, the advent of combined PET/CT scanners in clinical imaging practice has brought its own specific pitfalls and artefacts. A knowledge of the normal distribution of FDG and its physiological variation is essential before interpreting PET scans as well as an awareness of the potential false positive and negative cases that can occur. It is important that referring clinicians, PET/CT interpreters and imaging technologists/radiographers are aware of potential pitfalls so that their impact is minimised and that the image data are acquired and interpreted in the most accurate manner. With careful patient preparation, attention to detail and adequate training, the se artefacts and pitfalls may be minimised allowing this powerful hybrid imaging technique to realise it's potential. This paper attempts to describe some of the common pitfalls and artefacts and how they can be avoided or appropriately interpreted. PMID- 17464271 TI - Perioperative methemoglobinemia. AB - We report a case of a male patient who underwent splenectomy for idiopathic thrombocytopenic purpura. Oxygen saturation was 92% by pulse oximetry and cyanosis was apparent during the perioperative period. Methemoglobin (metHb) levels were 10.4%. After removal of the tracheal tube, intravenous methylene blue (1 mg/kg) was administered because of persistently low SpO2 levels (87%). During preanesthetic evaluation, the patient did not inform the anesthesiologist that he had been taking dapsone. PMID- 17464272 TI - Clinical controversies surrounding the diagnosis and treatment of hepatopulmonary syndrome. AB - Hepatopulmonary syndrome (HPS) is a clinical syndrome which complicates liver disease but remains poorly defined. To date there is no consensus on the definition of hypoxemia or the best diagnostic test that provides evidence of intrapulmonary shunting. Furthermore, it is reasonable to question whether the role of intrapulmonary shunting has been overplayed in the etiology of HPS and if echocardiographic intrapulmonary shunts could simply be nonspecific markers of the hyperdynamic circulation of liver disease. These inquiries lend support to theories that HPS is primarily a manifestation of profound pulmonary vascular autoregulatory failure that occurs in predisposed patients. Because there is little experience with clinical interventions for HPS other than liver transplantation, there are few accepted, efficacious therapeutic options for these patients. Because liver transplantation remains the only treatment known to effectively ''treat'' HPS, many medical communities have increased the priority of HPS patients over others who are waiting for life saving organs. Until there is further evidence that effectively identifies patients with HPS and that predicts outcomes, the preferential allocation of organs to patients with suspected HPS may unjustly cause the death of others waiting for transplantation. PMID- 17464273 TI - Back posture education in elementary schoolchildren: stability of two-year intervention effects. AB - AIM: The study's first objective was to evaluate class teachers' efforts to promote good body mechanics after a structured back education program was finished and to evaluate whether their support during follow-up resulted in better intervention effects at 1-year follow-up. Secondary, the stability of intervention effects on children's back posture knowledge, fear-avoidance beliefs and back pain reports following a 2-school-year multi-factorial back education program was evaluated at 1-year follow-up. An additional focus was put on what young children learned about good body mechanics in the obligatory school curriculum compared to intensive back posture promotion. METHODS: The quasi experimental study included at baseline 398 elementary schoolchildren aged 8-11 years. The back education program consisted of 13 h back education and the stimulation of postural dynamism in the class through support and environmental changes lasting 2 school-years. Controls received the obligatory curriculum, not including back education. Evaluation consisted of a questionnaire, which was filled out by 121 intervention children and 124 controls at baseline, post-test and follow-up. Teachers were interviewed at the end of the follow-up school-year. RESULTS: Teachers continued with initiatives to increase postural dynamism in the class when they had been instructed about that matter. However, teachers' efforts to continue the promotion of good body mechanics showed no additional effect on children's knowledge. Improved back posture knowledge demonstrated stability at 1 year follow-up. Whereas the obligatory curriculum provided children with fundamental postural knowledge, the back posture program added important aspects. Fear-avoidance beliefs and self-reported pain were not increased at 1-year follow up. CONCLUSION: The stable intervention effects point out that intensive implementation of a structured multifactorial back education program in the elementary school curriculum is effective. PMID- 17464274 TI - Radiolabelled aptamers for tumour imaging and therapy. AB - Advances in biotechnology have led to new techniques for the design, selection and production of ligands suitable for molecular targeting. One promising approach is the production of specific receptor binding molecules based on defined nucleic acid sequences that are capable of recognising a wide array of target molecules. These oligonuclide ligands are known as aptamers. The technology that allows production of aptamer molecules is known as systematic evolution of ligands by exponential enrichment (SELEX). Using these techniques, aptamers can now be produced rapidly, inexpensively, and with high homogeneity. Furthermore, they are stable over long term storage at ambient room temperatures. A monomeric aptamer is small in size, with a molecular weight as low as 5 to 10 kDa. However, the aptamer molecule may be used as building block for custom designed targeting agents, offering several advantages. These molecules penetrate tumour readily, reach peak levels quickly and clear from the body rapidly, thus having properties of low toxicity and immunoreactivity. Previous work with radiolabelled aptamers is limited and is currently restricted to preclinical studies, but the body of evidence is steadily growing and aptamers are emerging as valuable clinical products for diagnostic imaging and therapy. We have shown that aptamers directed against the mucin 1 (MUC1) antigen, a tumour marker previously extensively used in tumour imaging and therapy, demonstrated high specificity and uniform penetration in tumour xenografts. The future strategy will be to manipulate the molecular weight of the molecules to achieve an optimum balance between the low immunogenicity and excellent tumour penetration for diagnostic imaging and targeted therapy. In this way, a balance can be achieved between the rapid renal clearance and adequate tumour uptake required for diagnostic imaging and targeted therapy. PMID- 17464275 TI - Development of anti-MUC1 di-scFvs for molecular targeting of epithelial cancers, such as breast and prostate cancers. AB - Pretargeted radioimmunotherapy (RIT) is a promising approach to increase the therapeutic index of RIT for malignant solid tumors. For pretargeted RIT of epithelial cancers, such as breast and prostate, mucin 1 (MUC1), the epithelial mucin, was chosen as a target antigen (Ag). Overexpression, hypoglycosylation and loss of apical distribution on the cellular membrane distinguish the tumor associated MUC1 from normal MUC1. These characteristics of MUC1, best known in breast cancer, were validated in prostate cancer. The multivalent bispecific MUC1 pretargeting molecule under development consists of a tumor binding module and a radioactive hapten capturing module. The building blocks of each module were chosen as single chain antibody fragments (scFv) to be covalently attached to a multifunctional polyethylene glycol (PEG) scaffold. PEGylation studies with scFvs selected from anti-MUC1 libraries and engineered with a free thiol for site specific conjugation showed that highest reaction yields were obtained with short monofunctional PEG molecules. To accommodate the use of a bifunctional PEG for covalent assembly of binding and capturing modules, the MUC1 binding module was developed into a di-scFv-SH format and optimized for linker length and location of the free thiol in respect to Ag binding and site-specific conjugation. Approaches under study to improve PEGylation yields with bifunctional PEG molecules include alkyne-azide cycloaddition. Assembly efficiencies, through PEGylation, of the binding and capturing modules and pharmacokinetics will influence the final valency of the MUC1 pretargeting molecule: anti-MUC1 di-scFv PEG- anti-radioactive hapten scFv or di-scFv-PEG-anti-radioactive hapten di-scFv. PMID- 17464276 TI - Use of a 99mTc labeled anti-TNFalpha monoclonal antibody in Crohn's disease: in vitro and in vivo studies. AB - AIM: Crohn's disease (CD) is a chronic inflammatory bowel disease characterized by a cellular-mediated immune response driven by cytokines secreted mainly by T helper 1 cells (Th1). In active phases of the disease, an increased production and release of tumor necrosis factor a (TNFalpha) by macrophages and monocytes of the lamina propria has been described. The aim of this study was to detect the presence of TNFalpha within the gut mucosa in patients with active CD by using (99m)Tc-labelled chimeric human/mouse monoclonal antibody anti-TNFalpha (Infliximab, Remicade). METHODS: Infliximab has been labeled with (99m)Tc after reduction of disulfide bound by 2-ME method. In vitro binding assay and biodistribution in animal of [(99m)Tc]Infliximab has been performed to evaluate the retention of its biological activity. Ten patients with active CD refractory to conventional medical therapies were studied. Images of the abdomen were acquired at 6 to 20 h after i.v. injection of about 10 mCi of [(99m)Tc]Infliximab and a week later, all patients were also studied with [(99m)Tc]HMPAO-labeled autologous white blood cells (WBC). RESULTS: A product with high labeling efficiency (>95%) and stability has been obtained. In vitro tests with stimulated T-cells expressing TNFalphalpha indicated that [(99m)Tc] Infliximab retains its binding activity to cell bound TNFalpha as compared to unlabelled Infliximab. The degree of [(99m)Tc]Infliximab uptake by the inflamed bowel evaluated at 20 h postinjection was much less than that seen with labeled WBC and with a different distribution. Three of these patients received anti-TNFalpha (Infliximab) for therapeutic purposes with good clinical results despite the scintigraphy with (99m)Tc-Infliximab was negative in 2 of them. CONCLUSION: Scintigraphy with [(99m)Tc]Infliximab shows the presence of little TNFalpha in the affected bowel of patients with active CD. Therefore, the clinical benefit that patients have from Infliximab therapy is unlikely the consequence of a local a reduction of TNFalpha and the mechanism of action of Infliximab, in therapeutic doses, deserves further investigations. PMID- 17464277 TI - Evaluation of [(111/114m)In]CHX-A''-DTPA-ZHER2:342, an affibody ligand coniugate for targeting of HER2-expressing malignant tumors. AB - AIM: Radionuclide imaging of the HER2 receptor, which is a target for trastuzumab therapy, can provide important diagnostic information. Further, targeting radionuclide therapy might be an option for treatment of HER2 expressing tumors. The phage-display selected Affibody ligand Z(HER2:342), which binds to HER2 with an affinity of 22 pM, may here play an important role. The small size of the Z(HER2:342), 7.5 kDa, enables quick tumor localization and fast blood clearance. Earlier, successful targeting of HER2-expressing xenografts using Z(HER2:342) labeled using [(111)In]benzyl-DTPA was reported. By changing to the CHX-A''-DTPA chelator, the stability and labeling kinetics of the radiometal-Z(HER2:342) conjugate can be improved. The aim of this study was to evaluate the labeling of the CHX-A''-DTPA-Z(HER2:342) conjugate with (111)In for diagnostic imaging and with (114m)In for locoregional radionuclide therapy. METHODS: The isothiocyanate derivative of CHX-A''-DTPA was coupled to Z(HER2:342) in alkaline conditions at 37 degrees C. The conjugate was labeled with both (111)In and (114m)In and evaluated in vitro and in vivo. RESULTS: Labeling with (111)In and (114m)In provided >95% yield after 30 min at RT. Specific radioactivity was 0.5 and 12 MBq/nmol, for (114m)In and (111)In, respectively. The radiolabeled conjugates demonstrated specific binding to HER2 expressing SKOV-3 cells. In mice bearing SKOV-3 xenografts, the tumor uptake of [(111)In]CHX-A''-DTPA-Z(HER2:342) 4 h postinjection was 10.3+/-3.6% IA/g and tumor-to-blood ratio about 190. CONCLUSION: [(111)In]CHX-A''-DTPA-Z(HER2:342) is a promising candidate for the visualization of HER2 expression in malignant tumors. Labeled with (114m)In it could also be used for locoregional treatment of HER2 expressing tumors. PMID- 17464278 TI - Estimation of metastatic cure after radionuclide therapy. AB - Targeted radionuclide therapy of disseminated tumor disease involves many factors that will affect the therapeutic outcome. For optimization of such therapy, it is important to know how these factors affect the therapeutic outcome. In this paper, the metastatic cure probability (MCP) model is described. The MCP model is a valuable tool for analyses of the various factors influencing the metastatic cure. The factors discussed are: 1) the physical parameters (the energy and range of emitted particles, absorbed dose-distributions, and cross-irradiation of tumors from the surrounding normal tissue) and 2) the biological parameters (radiosensitivity of tumors, tumor distributions, tumor growth rate, metastasis formation rate, variable tumor activity concentration and non-homogeneous tumor activity distributions). PMID- 17464279 TI - Has radio-targeting come of age? Report of the 18th IRIST meeting. PMID- 17464280 TI - Fixed obstructive lung disease among workers in the flavor-manufacturing industry -California, 2004-2007. AB - Bronchiolitis obliterans, a rare and life-threatening form of fixed obstructive lung disease, is known to be caused by exposure to noxious gases in occupational settings and has been described in workers in the microwave-popcorn industry who were exposed to artificial butter-flavoring chemicals, including diacetyl. In August 2004, the California Department of Health Services (CDHS) and Division of Occupational Safety and Health (Cal/OSHA) received the first report of a bronchiolitis obliterans diagnosis in a flavor-manufacturing worker in California. In April 2006, a second report was received of a case in a flavor manufacturing worker from another company. Neither worker was employed in the microwave-popcorn industry; both were workers in the flavor-manufacturing industry, which produces artificial butter flavoring and other flavors such as cherry, almond, praline, jalapeno, and orange. Both workers had handled pure diacetyl, an ingredient in artificial butter and other flavorings, and additional chemicals involved in the manufacturing process. Studies have indicated that exposure to diacetyl causes severe respiratory epithelial injury in animals. Because the manufacture of flavorings involves more than 2,000 chemicals, workers in the general flavor-manufacturing industry are exposed to more chemicals than workers in the microwave-popcorn industry, which primarily uses butter flavorings. Food flavorings are designated "generally recognized as safe" when approved by the U.S. Food and Drug Administration; flavorings are not known to put consumers at risk for lung disease. This report describes the first two cases of bronchiolitis obliterans in flavor-manufacturing workers in California, the findings of the public health investigation, and the actions taken by state and federal agencies to prevent future cases of occupational bronchiolitis obliterans. To identify cases and reduce risk for lung disease from occupational exposure to flavorings, a timely, effective response is needed, including medical surveillance, exposure monitoring, and reduced exposure. PMID- 17464281 TI - Nonfatal occupational injuries and illnesses--United States, 2004. AB - Data collected through a National Electronic Injury Surveillance System occupational supplement (NEISS-Work) provide information on persons treated for nonfatal work-related injuries and illnesses in U.S. hospital emergency departments (EDs). CDC's National Institute for Occupational Safety and Health uses these data to monitor injury trends and aid prevention activities. This report summarizes 2004 NEISS-Work injury and illness surveillance data. In 2004, an estimated 3.4 million nonfatal ED-treated injuries and illnesses occurred among workers of all ages, with a rate of 2.5 cases per 100 full-time equivalent (FTE) workers aged > or =15 years. Workers aged <25 years had the highest injury/illness rates. More than three fourths of all nonfatal workplace injuries/illnesses were attributed to contact with objects or equipment (e.g., being struck by a falling tool or caught in machinery), bodily reaction or exertion (e.g., a sprain or strain), and falls. No substantial reduction was observed in the overall number and rate of ED-treated occupational injuries/illnesses during 1996-2004. To reduce occupational injuries/illnesses, interventions should continue to target workers at highest risk and reduce exposure to those workplace hazards with the greatest potential for causing severe injury or death. More emphasis should be placed on prevention effectiveness studies and dissemination of successful interventions to reduce work-related injuries and illnesses. PMID- 17464282 TI - Lead exposure among females of childbearing age--United States, 2004. AB - For centuries, exposure to high concentrations of lead has been known to pose health hazards, and evidence is mounting regarding adverse health effects from moderate- and low-level blood lead concentrations. Public health authorities use higher levels to define blood lead levels (BLLs) of concern in nonpregnant females (> or =25 microg/dL) compared with children (> or =10 microg/dL) and a lower level (> or =5 microg/dL) for pregnant females. This difference in levels for nonpregnant and pregnant females has raised concern because of the recognition that a proportion of nonpregnant females with BLLs > or =5 microg/dL will become pregnant and potentially expose their infants to a risk for adverse health effects from lead. Maternal and fetal BLLs are nearly identical because lead crosses the placenta unencumbered. This report summarizes 2004 surveillance data regarding elevated BLLs among females of childbearing age (i.e., aged 16-44 years) in 37 states participating in CDC's Adult Blood Lead Epidemiology and Surveillance (ABLES) program. The results indicated that rates of elevated BLLs ranged from 0.06 per 100,000 females of childbearing age at BLLs of > or =40 microg/dL to 10.9 per 100,000 females at BLLs of > or =5 microg/dL. Primary and secondary prevention of lead exposure among females of childbearing age is needed to avert neurobehavioral and cognitive deficits in their offspring. PMID- 17464283 TI - Spinophilin/neurabin reciprocally regulate signaling intensity by G protein coupled receptors. AB - Spinophilin (SPL) and neurabin (NRB) are structurally similar scaffolding proteins with several protein binding modules, including actin and PP1 binding motifs and PDZ and coiled-coil domains. SPL also binds regulators of G protein signaling (RGS) proteins and the third intracellular loop (3iL) of G protein coupled receptors (GPCRs) to reduce the intensity of Ca(2+) signaling by GPCRs. The role of NRB in Ca(2+) signaling is not known. In the present work, we used biochemical and functional assays in model systems and in SPL(-/-) and NRB(-/-) mice to show that SPL and NRB reciprocally regulate Ca(2+) signaling by GPCRs. Thus, SPL and NRB bind all members of the R4 subfamily of RGS proteins tested (RGS1, RGS2, RGS4, RGS16) and GAIP. By contract, SPL, but not NRB, binds the 3iL of the GPCRs alpha(1B)-adrenergic (alpha(1B)AR), dopamine, CCKA, CCKB and the muscarinic M3 receptors. Coexpression of SPL or NRB with the alpha(1B)AR in Xenopus oocytes revealed that SPL reduces, whereas NRB increases, the intensity of Ca(2+) signaling by alpha(1B)AR. Accordingly, deletion of SPL in mice enhanced binding of RGS2 to NRB and Ca(2+) signaling by alphaAR, whereas deletion of NRB enhanced binding of RGS2 to SPL and reduced Ca(2+) signaling by alphaAR. This was due to reciprocal modulation by SPL and NRB of the potency of RGS2 to inhibit Ca(2+) signaling by alphaAR. These findings suggest a novel mechanism of regulation of GPCR-mediated Ca(2+) signaling in which SPL/NRB forms a functional pair of opposing regulators that modulates Ca(2+) signaling intensity by GPCRs by determining the extent of inhibition by the R4 family of RGS proteins. PMID- 17464284 TI - Autoprocessing of the Vibrio cholerae RTX toxin by the cysteine protease domain. AB - Vibrio cholerae RTX is a large multifunctional bacterial toxin that causes actin crosslinking. Due to its size, it was predicted to undergo proteolytic cleavage during translocation into host cells to deliver activity domains to the cytosol. In this study, we identified a domain within the RTX toxin that is conserved in large clostridial glucosylating toxins TcdB, TcdA, TcnA, and TcsL; putative toxins from V. vulnificus, Yersinia sp., Photorhabdus sp., and Xenorhabdus sp.; and a filamentous/hemagglutinin-like protein FhaL from Bordetella sp. In vivo transfection studies and in vitro characterization of purified recombinant protein revealed that this domain from the V. cholerae RTX toxin is an autoprocessing cysteine protease whose activity is stimulated by the intracellular environment. A cysteine point mutation within the RTX holotoxin attenuated actin crosslinking activity suggesting that processing of the toxin is an important step in toxin translocation. Overall, we have uncovered a new mechanism by which large bacterial toxins and proteins deliver catalytic activities to the eukaryotic cell cytosol by autoprocessing after translocation. PMID- 17464285 TI - Splicing factors stimulate polyadenylation via USEs at non-canonical 3' end formation signals. AB - The prothrombin (F2) 3' end formation signal is highly susceptible to thrombophilia-associated gain-of-function mutations. In its unusual architecture, the F2 3' UTR contains an upstream sequence element (USE) that compensates for weak activities of the non-canonical cleavage site and the downstream U-rich element. Here, we address the mechanism of USE function. We show that the F2 USE contains a highly conserved nonameric core sequence, which promotes 3' end formation in a position- and sequence-dependent manner. We identify proteins that specifically interact with the USE, and demonstrate their function as trans acting factors that promote 3' end formation. Interestingly, these include the splicing factors U2AF35, U2AF65 and hnRNPI. We show that these splicing factors not only modulate 3' end formation via the USEs contained in the F2 and the complement C2 mRNAs, but also in the biocomputationally identified BCL2L2, IVNS and ACTR mRNAs, suggesting a broader functional role. These data uncover a novel mechanism that functionally links the splicing and 3' end formation machineries of multiple cellular mRNAs in an USE-dependent manner. PMID- 17464286 TI - A guanine nucleobase important for catalysis by the VS ribozyme. AB - A guanine (G638) within the substrate loop of the VS ribozyme plays a critical role in the cleavage reaction. Replacement by any other nucleotide results in severe impairment of cleavage, yet folding of the substrate is not perturbed, and the variant substrates bind the ribozyme with similar affinity, acting as competitive inhibitors. Functional group substitution shows that the imino proton on the N1 is critical, suggesting a possible role in general acid-base catalysis, and this in accord with the pH dependence of the reaction rate for the natural and modified substrates. We propose a chemical mechanism for the ribozyme that involves general acid-base catalysis by the combination of the nucleobases of guanine 638 and adenine 756. This is closely similar to the probable mechanism of the hairpin ribozyme, and the active site arrangements for the two ribozymes appear topologically equivalent. This has probably arisen by convergent evolution. PMID- 17464287 TI - Structural basis of the 3'-end recognition of a leading strand in stalled replication forks by PriA. AB - In eubacteria, PriA helicase detects the stalled DNA replication forks. This critical role of PriA is ascribed to its ability to bind to the 3' end of a nascent leading DNA strand in the stalled replication forks. The crystal structures in complexes with oligonucleotides and the combination of fluorescence correlation spectroscopy and mutagenesis reveal that the N-terminal domain of PriA possesses a binding pocket for the 3'-terminal nucleotide residue of DNA. The interaction with the deoxyribose 3'-OH is essential for the 3'-terminal recognition. In contrast, the direct interaction with 3'-end nucleobase is unexpected, considering the same affinity for oligonucleotides carrying the four bases at the 3' end. Thus, the N-terminal domain of PriA recognizes the 3'-end base in a base-non-selective manner, in addition to the deoxyribose and 5'-side phosphodiester group, of the 3'-terminal nucleotide to acquire both sufficient affinity and non-selectivity to find all of the stalled replication forks generated during DNA duplication. This unique feature is prerequisite for the proper positioning of the helicase domain of PriA on the unreplicated double stranded DNA. PMID- 17464288 TI - Interdomain association in fibronectin: insight into cryptic sites and fibrillogenesis. AB - The process by which fibronectin (FN), a soluble multidomain protein found in tissue fluids, forms insoluble fibrillar networks in the extracellular matrix is poorly understood. Cryptic sites found in FN type III domains have been hypothesized to function as nucleation points, thereby initiating fibrillogenesis. Exposure of these sites could occur upon tension-mediated mechanical rearrangement of type III domains. Here, we present the solution structures of the second type III domain of human FN ((2)FNIII), and that of an interaction complex between the first two type III domains ((1-2)FNIII). The two domains are connected through a long linker, flexible in solution. A weak but specific interdomain interaction maintains (1-2)FNIII in a closed conformation that associates weakly with the FN N-terminal 30 kDa fragment (FN30 kDa). Disruption of the interdomain interaction by amino-acid substitutions dramatically enhances association with FN30 kDa. Truncation analysis of (1 2)FNIII reveals that the interdomain linker is necessary for robust (1-2)FNIII FN30 kDa interaction. We speculate on the importance of this interaction for FN function and present a possible mechanism by which tension could initiate fibrillogenesis. PMID- 17464289 TI - Structure of colicin I receptor bound to the R-domain of colicin Ia: implications for protein import. AB - Colicin Ia is a 69 kDa protein that kills susceptible Escherichia coli cells by binding to a specific receptor in the outer membrane, colicin I receptor (70 kDa), and subsequently translocating its channel forming domain across the periplasmic space, where it inserts into the inner membrane and forms a voltage dependent ion channel. We determined crystal structures of colicin I receptor alone and in complex with the receptor binding domain of colicin Ia. The receptor undergoes large and unusual conformational changes upon colicin binding, opening at the cell surface and positioning the receptor binding domain of colicin Ia directly above it. We modelled the interaction with full-length colicin Ia to show that the channel forming domain is initially positioned 150 A above the cell surface. Functional data using full-length colicin Ia show that colicin I receptor is necessary for cell surface binding, and suggest that the receptor participates in translocation of colicin Ia across the outer membrane. PMID- 17464292 TI - Suitability of hCMV for viral gene expression in the brain. PMID- 17464290 TI - Myc targets Cks1 to provoke the suppression of p27Kip1, proliferation and lymphomagenesis. AB - Reduced levels of the cyclin-dependent kinase inhibitor p27(Kip1) connote poor prognosis in cancer. In human Burkitt lymphoma and in precancerous B cells and lymphomas arising in Emu-Myc transgenic mice, p27(Kip1) expression is markedly reduced. We show that the transcription of the Cks1 component of the SCF(Skp2) complex that is necessary for p27(Kip1) ubiquitylation and degradation is induced by Myc. Further, Cks1 expression is elevated in precancerous Emu-Myc B cells, and high levels of Cks1 are also a hallmark of Emu-Myc lymphoma and of human Burkitt lymphoma. Finally, loss of Cks1 in Emu-Myc B cells elevates p27(Kip1) levels, reduces proliferation and markedly delays lymphoma development and dissemination of disease. Therefore, Myc suppresses p27(Kip1) expression, accelerates cell proliferation and promotes tumorigenesis at least in part through its ability to selectively induce Cks1. PMID- 17464293 TI - Splitting or stacking fluorescent proteins to visualize mRNA in living cells. PMID- 17464295 TI - The vanilloid receptor TRPV1: 10 years from channel cloning to antagonist proof of-concept. AB - The clinical use of TRPV1 (transient receptor potential vanilloid subfamily, member 1; also known as VR1) antagonists is based on the concept that endogenous agonists acting on TRPV1 might provide a major contribution to certain pain conditions. Indeed, a number of small-molecule TRPV1 antagonists are already undergoing Phase I/II clinical trials for the indications of chronic inflammatory pain and migraine. Moreover, animal models suggest a therapeutic value for TRPV1 antagonists in the treatment of other types of pain, including pain from cancer. We argue that TRPV1 antagonists alone or in conjunction with other analgesics will improve the quality of life of people with migraine, chronic intractable pain secondary to cancer, AIDS or diabetes. Moreover, emerging data indicate that TRPV1 antagonists could also be useful in treating disorders other than pain, such as urinary urge incontinence, chronic cough and irritable bowel syndrome. The lack of effective drugs for treating many of these conditions highlights the need for further investigation into the therapeutic potential of TRPV1 antagonists. PMID- 17464296 TI - Targeting polyamine metabolism and function in cancer and other hyperproliferative diseases. AB - The polyamines spermidine and spermine and their diamine precursor putrescine are naturally occurring, polycationic alkylamines that are essential for eukaryotic cell growth. The requirement for and the metabolism of polyamines are frequently dysregulated in cancer and other hyperproliferative diseases, thus making polyamine function and metabolism attractive targets for therapeutic intervention. Recent advances in our understanding of polyamine function, metabolic regulation, and differences between normal cells and tumour cells with respect to polyamine biology, have reinforced the interest in this target-rich pathway for drug development. PMID- 17464297 TI - Diplopia following cataract surgery: a review of 150 patients. AB - AIM: To study the motility pattern, underlying mechanism, and management of patients who complained of double vision after cataract surgery. METHODS: A retrospective case note analysis of 150 patients presenting with diplopia after cataract surgery to an orthoptic clinic over a 70-month period. Information was retrieved from orthoptic, ophthalmological, and operating room records. RESULTS: A total of 3% of patients presenting to the orthoptic clinic had diplopia after cataract surgery. We grouped these according to the underlying mechanisms which were: (1) decompensating pre-existing strabismus (34%), (2) extraocular muscle restriction/paresis (25%), (3) refractive (8.5%), (4) concurrent onset of systemic disease (5%), (5) central fusion disruption (5%), and (6) monocular diplopia (2.5%). Twenty per cent of the patients could not be categorised with certainty. After infiltrational anaesthesia, extraocular muscle restriction/paresis was the commonest presentation, while decompensation of preexisting strabismus was commonest with topical anaesthesia.For the 150 patients seen, prisms were the commonest form of treatment prescribed (64%) either in isolation or in combination with other treatment, including surgery (19%). Convergence and divergence insufficiency/paresis patterns were also common. A changing motility pattern was noted in some patients who had early documentation, with increasing comitance over time (spread of comitance). Partial resolution made it difficult to clearly identify the underlying mechanism in patients with late documentation. CONCLUSION: Double vision is a troublesome complication of otherwise successful cataract surgery. The use of topical anaesthesia does not abolish this surgical risk. PMID- 17464298 TI - Contact urticaria to ultrasonic gel. PMID- 17464299 TI - Combined phacoemulsification and deep sclerectomy (PDS) with intraoperative mitomycin C (MMC) augmentation. AB - AIM: To report the safety and efficacy of intraoperative mitomycin (MMC) augmentation of combined phacoemulsification and deep sclerectomy (PDS). METHODS: Retrospective, non-randomized, comparative, interventional case series of 119 eyes (63 with and 56 without MMC augmentation) of 119 patients who had PDS between September 2001 and April 2004. RESULTS: The mean follow-up was 23 months (range 12-41 months). There were no differences in the baseline characteristics of the two groups except that patients from the phacoemulsification and deep sclerectomy with mitomycin C (PDS-MMC) group were on average, younger by 3 years (P=0.01). Two years after surgery, the probability of maintaining an IOP below 19 and 15 mmHg without glaucoma medications or needle revision was 76 and 62% in the PDS-MMC group and 62 and 45% in the PDS-no MMC group (P=0.02 and 0.04, respectively). Nd:YAG laser goniopuncture was performed in 71.4% of eyes in the PDS-no MMC and 61.9% of the PD-MMC group (P=0.33). Needle revision was performed in 21.4% of the PDS-no MMC and 17.4% of the PDS-MMC group (P=0.65). Ten patients (8.4%) lost more than two lines of Snellen's visual acuity during follow-up, with no difference between the groups. There were few serious complications related to MMC use (hypotony in one eye after laser goniopuncture). The overall incidence of transconjunctival oozing in the PDS-MMC group was 9.5% compared with 5.4% in the PDS-no MMC group. CONCLUSION: This study demonstrates that augmentation of PDS with MMC is safe. MMC augmentation appears to increase the probability of achieving lower target intraocular pressures after combined PDS. PMID- 17464300 TI - Orbital schwannomas: findings from magnetic resonance imaging in 62 cases. AB - PURPOSE: To evaluate the diagnostic value of magnetic resonance imaging (MRI) and illustrate the common MRI features of orbital schwannoma. METHODS: We reviewed 62 consecutive cases of pathologically proven orbital schwannoma investigated by MRI. All cases were examined using T1- and T2-weighted images. Enhanced T1 weighted images were obtained after administration of gadopentetate dimeglumine. The images were analysed and compared. RESULTS: Cone-shaped lesions were the most frequent (16, 26%), followed by dumbbell-shaped (10, 16%), oval (9, 15%) and round lesions (8, 13%). The most common site was the superior aspect of the orbit (17, 27%), followed by the medial superior (12, 19%) and the orbital apex (12, 19%). On unenhanced T1-weighted images, 53 (85%) lesions showed isointensity or small patches of hypointensity. The patterns of enhancement seen on T1-weighted images correlated with the signal characteristics of unenhanced T2-weighted images. Five patterns emerged in our series. Among these patterns, the most common sign, found in 18 (29%) cases, was peripheral (ring) enhancement on enhanced T1-weighted images, which showed peripheral isointensity with central hyperintensity on T2-weighted images. Lesions with homogeneous or heterogeneous enhancement were seen in 16 (26%) and 26 (42%) cases, respectively.ConclusionsMRI is a valuable diagnostic method for orbital schwannoma, particularly when contrast is applied. The locations and shapes of tumours can be seen distinctly. T1-weighted images are relatively nonspecific. T2-weighted and enhanced T1 weighted images provide information about the pathology of tumours. In particular, peripheral enhancement should be considered a target sign of schwannoma.Eye (2008) 22, 1034-1039; doi:10.1038/sj.eye.6702832; published online 20 April 2007. PMID- 17464301 TI - Tonometer disinfection practice in the United Kingdom: a national survey. AB - PURPOSE: To assess current tonometer disinfection practice in the UK, and compare with published recommendations. METHODS: Every ophthalmology unit with training recognition in the UK was contacted (n=155). A senior nurse at each institution completed a telephone questionnaire regarding local tonometer disinfection practice. RESULTS: The response rate was 100%. Thirty-five units (23%) reported exclusive use of disposable tonometer heads and were excluded from further analysis. One hundred and twenty units (77%) used either reusable or a combination of reusable and disposable tonometer heads. Where reusable heads were used, 80 units (67%) immersed them in a chlorine-based solution such as sodium hypochlorite or sodium dichloroisocyanurate. Others used isopropyl alcohol (18 units), hydrogen peroxide (12 units), chloramine (5 units), chlorhexidine (4 units) and peracetic acid (1 unit). Where a chlorine-based agent was used, the concentration of available chlorine ranged from 125 to 30 000 p.p.m., with 50 units (63%) using a concentration of less than 5 000 p.p.m. (i.e., inadequate based on published recommendations). Where the tonometer head was immersed in disinfectant between patients (n=101), 29 units (29%) provided just one tonometer head per practitioner, making adequate soak time between patients unlikely. Every unit replenished the disinfectant at least daily, deemed sufficient for most agents. However, hydrogen peroxide solutions should be replenished twice daily, which did not take place in nine units. CONCLUSION: This survey reveals disparity between current tonometer disinfection practice and published international recommendations, with some institutions using practices that may render patients susceptible to transmissible infection. PMID- 17464302 TI - An analysis of the CFH Y402H genotype in AMD patients and controls from the UK, and response to PDT treatment. AB - AIM: Mutation in the complement factor H (CFH) gene is an important risk factor for age-related macular degeneration (AMD). In this study, we identified the strength of the CFH Y402H gene variant association in a UK AMD cohort and tested the hypothesis that this variant may influence the biological response of choroidal neovascularisation (CNV) following photodynamic therapy (PDT) for CNV. METHODS: A total of 557 cases with AMD and 551 normal controls were genotyped for the CFH Y402H (1277 C/T) variant using the 5' nuclease TaqMan assay for allelic discrimination. The CFH gene association for AMD, for the different CNV subtypes and for patients needing PDT was estimated. Twenty-seven PDT-treated patients were followed up for 15 months with ETDRS-derived vision, clinical examination, and fundus angiography. Individuals with different CFH genotypes were then analysed for any association with visual change following PDT. RESULTS: The risk association for AMD with the CFH CC genotype (odd ratio (OR)=3.62, Pc<0.0001) was similar to that reported in other Caucasian cohorts. The magnitude and strength of this association was stronger in AREDS stages 2-4 (ORs=4.48, 2.69, and 5.17). ORs for the risk of predominantly classic CNV were significantly raised for both the CC (OR=17.87, P<0.0001) and CT (OR=9.06, P=0.0002) genotypes. The number of patients carrying the high-risk C allele was 70.4% in those requiring PDT as compared to 52.3% in the non-PDT group (OR=2.16, P=0.011), and presence of the CC genotype significantly increased the risk of PDT (OR=5.48, P=0.015). The degree of visual loss following PDT was significantly higher in the CFH CC genotype group (P=0.038); 50% of CC cases (n=13) and 45% of the CT cases (n=12) lost 15 or more ETDRS letters at final follow-up. CONCLUSION: In this UK cohort of AMD patients, the CFH Y402H variant was significantly enriched in patients with predominantly classic CNV. Patients homozygous for the CFH Y402H genotype seem to have worse visual acuity after PDT. PMID- 17464303 TI - Outcomes following 25-gauge vitrectomies. AB - PURPOSE: Twenty-five gauge vitrectomy surgery offers potential advantages over standard 20-gauge vitrectomy surgery, but the short- and long-term post-operative complications, such as cataract formation, are still being evaluated. This study quantifies the outcomes seen following 25-gauge vitrectomies. METHODS: This is a retrospective, consecutive, non-comparative case series of 25-gauge vitrectomies performed between January 2002 and August 2004. Cases without at least 3 months of follow-up and previous vitrectomies were excluded. Analyses were performed with t-test and Kaplan-Meier curves. RESULTS: Seventy-one cases met inclusion criteria. The mean age of the patients was 65 years old (SD 11 years). A variety of surgical indications were included. A statistically significant difference was seen between the mean preoperative visual acuity (20/100) and the mean visual acuity at the 3-month post-operative visit (20/60; P<0.0001). A Kaplan-Meier curve established that for all cases 63.4% of eyes required cataract extraction at 1 year. Total mean follow-up time was 8.6+/-5.5 months. CONCLUSIONS: Statistically significant improvement was seen in mean vision by 3 months following 25-gauge vitrectomy. Cataract formation after 25-gauge vitrectomies remains an important consideration. PMID- 17464304 TI - Relation of optical coherence tomography and unusual angiographic leakage patterns in central serous chorioretinopathy. AB - PURPOSE: To investigate the anatomic basis of atypical angiographic leaks in central serous chorioretinopathy (CSC) with optical coherence tomography (OCT). METHODS: Fluorescein angiography (FA) and OCT were performed in three eyes of three patients (two men, one woman) with CSC. The angiographic leaks were treated with transpupillary thermotherapy (TTT) in two patients with long-standing CSC. The investigations were repeated in the treated patients during follow-up visits. RESULTS: Clinically, all patients demonstrated typical CSC; the female patient had subretinal fibrin under the detachment. FA showed unusual leakage patterns and OCT revealed bridging tissue connecting the pigment epithelial detachment (PED) to the overlying detached retina in all patients. CSC resolved completely in the two patients who underwent TTT along with normalization of the OCT findings. In one patient re-evaluated before complete resolution of CSC, OCT showed a flattened PED with disappearance of the bridging tissue and persistent serous detachment. FA demonstrated conversion of the previously atypical leak into a classic 'smokestack' configuration. Over the next month, leakage resolved completely. CSC and the anatomical findings persisted in the untreated patient. CONCLUSION: OCT identified a potential anatomic basis for unusual angiographic leakage pattern in all three cases of CSC evaluated. PMID- 17464305 TI - Anterior capsule cover and axial movement of intraocular lens. AB - PURPOSE: To measure optic shift (OS) of a single piece monofocal intraocular lens (IOL) with varying relationships between anterior capsule cover and IOL optic. METHODS: This is a prospective randomized masked study of 150 eyes undergoing phacoemulsification. Eyes received either option: 360 degrees total capsule cover (group I); partial cover (group II); or no cover (group III). OS was calculated as difference in anterior chamber depth after administration of cyclopentolate 1% and pilocarpine 2% on IOLMaster at separate visits at 6 months follow-up. Subsequently, using retro-illumination photographs, percentage area of capsule cover was calculated. OS within and between groups I and II was analyzed. The impact of one quartile change in area of capsule cover on percentage change in OS was measured for both groups. Unpaired t-test, correlation, and regression were applied. RESULTS: In groups I, II, and III, mean age of patients was 56.68+/ 6.38, 57.09+/-7.34, 59.15+/-6.35 years, respectively; mean OS (mm) was 1.25+/ 0.28, 1.20+/-0.24; 0.95+/-0.26 (P=0.013), respectively; and percentage area of capsule cover (%) was 47.35+/-10.48, 33.83+/-10.11, 0.16+/-0.13 (P=0.001), respectively. Mean percentage area of capsule cover in group I vsgroup II was significant (P=0.001). OS was 1.22+/-0.26 mm in groups I and II (combined) vs0.95+/-0.26 mm in group III (P=0.004, (0.06, 0.33)). OS in group I vsgroup II was not significant (P=0.46). Correlation coefficient was r=0.38 (P<0.001). With every increment on quartile (a quartile is any of the three values which divide the sorted data set into four equal parts, so that each part represents one fourth of the sample or population) class of area of capsule cover the OS increased by 0.12 mm. CONCLUSION: The OS differed significantly between total and partial cover groups combined vsno cover group. PMID- 17464306 TI - Perilimbal conjunctival pigmentation in Chinese patients with vernal keratoconjunctivitis. AB - PURPOSE: To document the disease spectrum and evaluate the presence of perilimbal conjunctival pigmentation in Chinese patients with vernal keratoconjunctivitis (VKC). METHOD: A case-control study was conducted between November 2004 and July 2005. Patients aged 18 or younger with VKC and age-matched children attending our eye clinic for refractive or orthoptic problems were recruited and compared. Detailed slit-lamp examination was performed noting in particular the presence of perilimbal conjunctival pigmentation, the severity of papillary reaction, and corneal complications of VKC. RESULTS: A total of 19 patients and 23 controls were evaluated. The presence of bilateral large tarsal or limbal papillae and epithelial defect were significantly associated with symptoms severity (Fisher's exact test, P=0.015 and P=0.035 respectively). All VKC patients were found to have perilimbal conjunctival pigmentation in at least one eye. There was a significant correlation in the colour and density of pigments between the two eyes (Sperman's rho=0.93, P<0.001). None of the controls was found to have such perilimbal conjunctival pigmentation (Fisher's exact test, P<0.001). CONCLUSION: The presence of perilimbal conjunctival pigmentation appears to be a consistent clinical finding in Chinese patients with VKC and may be a useful diagnostic sign for patients with subtle signs or symptoms. PMID- 17464307 TI - Orbital cellulitis associated with combined retinal and choroidal detachments. PMID- 17464308 TI - Intraocular use of rituximab. AB - PURPOSE: To evaluate the toxicity of 1 mg of intraocular rituximab and to present a small case-series of patients treated with intravitreal rituximab. METHODS: Rituximab (1 mg/0.1 ml) was injected in the vitreous of one eye of three Dutch belted rabbits. Two animals were injected with balanced salt solution as controls. At 1 month the rabbits were killed and the eyes examined by light microscopy. Three patients (five eyes) with intraocular lymphoma were also treated with a 1 mg injection of rituximab. RESULTS: The treated rabbit eyes and the control eyes showed no light microscopic evidence of ocular toxicity at 1 month following injection. The five human eyes of three patients have shown no evidence of intraocular toxicity with a median follow-up time of 3.6 months (range 2.0-6.4 months). One patient received a total of four injections in the right eye and three injections in the left eye. CONCLUSION: Intravitreal rituximab at a dose of 1 mg does not appear to cause toxicity in rabbit eyes and in the five eyes of three patients. PMID- 17464309 TI - Assessment of macular retinal thickness and volume in normal eyes and highly myopic eyes with third-generation optical coherence tomography. AB - PURPOSE: To compare the macular retinal thickness and macular volume between subjects with high myopia and non-myopia. METHODS: This prospective nonrandomized, comparative study recruited healthy subjects with high myopia subjects, defined as a spherical equivalence (SE) over -6 dioptres (D) or AXL>or=26.5 mm and the best corrected visual acuity better than 20/25, and subjects with non-myopia, defined as an with SE between 1.5D and -1.5 D and the BCVA better than 20/25. Optical coherence tomography was performed in each eye. RESULTS: Eighty high myopic eyes and 40 non-myopic eyes were included. The mean age of the high myopic group and non-myopia group was 29.6 and 27.5 years old, respectively. The mean refraction was -9.27 D in the high myopia group and -0.22 D in the non-myopia group. The high myopia group had significantly greater mean retinal thickness in the foveola and fovea 1 mm area than the non-myopia group (166 vs149 microm, P<0.0001, 199 vs188 microm, P=0.0063, respectively). However, the mean retinal thickness in the inner and outer macular area (superior, nasal, inferior, or temporal) of the high myopia group was significantly less than in the non-myopia group. In addition, the high myopia group had significantly smaller macular volume than the non-myopia group (P<0.0001). CONCLUSION: This study demonstrated that the retinal thickness in individuals with high myopia is thicker in the foveola and fovea, but thinner in the inner and the outer macular region. The retina of individuals with high myopia had smaller macular volume than those with non-myopia. PMID- 17464310 TI - Involvement of circulating endothelial progenitor cells and vasculogenic factors in the pathogenesis of diabetic retinopathy. PMID- 17464311 TI - DNA methylation in breast and colorectal cancers. AB - DNA methylation is one of several epigenetic changes observed in cells. Aberrant methylation of tumor suppressor genes, proto-oncogenes, and vital cell cycle genes has led many scientists to investigate the underlying cellular mechanisms of DNA methylation under normal and pathological conditions. Although DNA methylation is necessary for normal mammalian embryogenesis, both hypo- and hypermethylation of DNA are frequently observed in carcinogenesis and other pathological disorders. DNA hypermethylation silences the transcription of many tumor suppressor genes, resulting in immortalization of tumor cells. The reverse process, demethylation and restoration of normal functional expression of genes, is augmented by DNA methylation inhibitors. Recent studies suggest that DNA hypomethylation may also control gene expression and chromosomal stability. However, the roles of and relationship between hypomethylation and hypermethylation are not well understood. This review provides a brief overview of the mechanism of DNA methylation, its relationship to extrinsic stimulation including dietary intake and aging, and of abnormally methylated DNA in breast and colorectal cancers, which could be used as prognostic and diagnostic markers. PMID- 17464312 TI - Diffuse sclerosing variant of papillary thyroid carcinoma: lack of BRAF mutation but occurrence of RET/PTC rearrangements. AB - Diffuse sclerosing variant of papillary thyroid carcinoma (PTC) is a rare tumour with a characteristic morphology as well as a strong preponderance for younger female patients. The T1799A missense mutation in exon 15 of the BRAF gene and RET/PTC rearrangement have been identified as the dominant genetic tumour initiation events in the pathogenesis of PTC leading to a constitutive activation of the RAS-RAF-MAPK pathway. In order to elucidate the pathogenesis of diffuse sclerosing variant of PTC, the prevalence of BRAF mutation and RET/PTC were determined by RT-polymerase chain reaction and DNA-sequence analysis in tumour samples of seven patients with this variant (all female, age range 15-61 years, mean 33.3 years) without prior radiation exposure. None of these cases showed a BRAF mutation. RET/PTC1 (two out of seven) and RET/PTC3 (one out of seven), which have been shown in large PTC series to comprise together more than 90% of RET/PTC types, were found in <50% of the cases investigated. All seven samples expressed the RET tyrosine kinase domain but lacked its extracellular domain potentially suggesting the existence of rare types of RET/PTC rearrangement in the four remained cases of diffuse sclerosing variant of PTC. Regarding this subtype, our study confirmed the paradigm of a mutual exclusivity between RET/PTC and BRAF in PTC. Additionally, this rare variant of papillary thyroid carcinoma may represent a tumour type susceptible to RET-targeted therapies. PMID- 17464313 TI - MIB1 expression in basal cell layer: a diagnostic tool to identify premalignancies of the vulva. AB - Lichen sclerosus, high-grade usual vulvar intraepithelial neoplasia (VIN) and differentiated VIN have a different malignant potential. The objective of this study was to quantify the proliferative activity in the basal region of the epithelium of vulvar premalignancies. Furthermore, we investigated whether MIB1 expression in the basal region of vulvar epithelium can be helpful in diagnosing differentiated VIN, which may be hard to discern from normal epithelium. MIB1 was used to immunohistochemically visualise proliferating cells within formalin fixed, paraffin-embedded, archival tissue sections of different vulvar premalignancies (N=48) and normal vulvar epithelium (N=16). Automatic digital image analysis software was developed to quantify the proliferating fraction in different parts of the epithelium (MIB1 positivity index). MIB1 expression differed among the various vulvar premalignancies; a MIB1-negative basal cell layer was a distinct feature of normal vulvar epithelium. No MIB1-negative basal cell layer was noted in differentiated VIN or other vulvar premalignancies. Owing to this negative cell layer, the MIB1 proliferation index in normal vulvar epithelium was significantly lower than in vulvar premalignancies. In conclusion, MIB1 expression can be a helpful tool in diagnosing a premalignancy and has additional value especially to distinguish differentiated VIN neoplasia from normal vulvar epithelium, but cannot explain the differences in malignant potential. PMID- 17464314 TI - Primary pulmonary and mediastinal synovial sarcoma: a clinicopathologic study of 60 cases and comparison with five prior series. AB - Primary pulmonary and mediastinal synovial sarcoma is rare and poses a diagnostic challenge particularly when unusual histological features are present. We present 60 cases of primary pulmonary and mediastinal synovial sarcoma (29 male and 27 female subjects; mean age, 42 years) and compare our results with five prior series to better define unusual histological features. Clinically, patients with mediastinal synovial sarcoma were younger with a male gender bias. Radiologically, tumors were well delineated with distinctive magnetic resonance imaging features and little vascular enhancement. In all, 21/46 patients died of disease within 5 years. Histologically, all tumors had dense cellularity, interlacing fascicles, hyalinized stroma, and mast cell influx. Hemangiopericytoma-like vasculature (48/60), focal myxoid change (30/60), and entrapped pneumocytes (23/60) were seen. Calcification was not prevalent (10/60). Unusual histological features included Verocay body-like formations (7/60), vague rosettes (6/60), well-formed papillary structures (3/60), adenomatoid change (3/60), and rhabdoid morphology (2/60). Immunohistochemistry demonstrated expression of pancytokeratin (39/58), epithelial membrane antigen (29/53), cytokeratin 7 (26/40), cytokeratin 5/6 (5/7), calretinin (15/23), CD99 (19/23), bcl-2 (24/24), CD56 (11/11), S-100 (9/51), and smooth muscle actin (8/32). In total, 92% (36/39) of primary pulmonary and mediastinal synovial sarcomas studied were positive for t(x;18). In conclusion, our study confirms the clinical, histological, immunohistochemical, and molecular data from previous large series of primary pulmonary and mediastinal synovial sarcoma. Compared with soft tissue synovial sarcoma, primary pulmonary and mediastinal synovial sarcoma has less calcification, less obvious mast cell influx, and less radiologic vascularity, but similar magnetic resonance imaging features, percentage of poorly differentiated tumors, and number of t(x;18)-positive tumors. Awareness of focal unusual histology can prevent misdiagnosis particularly in t(x;18)-negative tumors. PMID- 17464315 TI - Gene expression analysis of soft tissue sarcomas: characterization and reclassification of malignant fibrous histiocytoma. AB - In soft tissue sarcomas, the diagnosis of malignant fibrous histiocytoma (MFH) has been a very controversial issue, and MFH is now considered to be reclassified into pleomorphic subtypes of other sarcomas. To characterize MFH genetically, we used an oligonucleotide microarray to analyze gene expression in 105 samples from 10 types of soft tissue tumors. Spindle cell and pleomorphic sarcomas, such as dedifferentiated liposarcoma, myxofibrosarcoma, leiomyosarcoma, malignant peripheral nerve sheath tumor (MPNST), fibrosarcoma and MFH, showed similar gene expression patterns compared to other tumors. Samples from those five sarcoma types could be classified into respective clusters based on gene expression by excluding MFH samples. We calculated distances between MFH samples and other five sarcoma types (dedifferentiated liposarcoma, myxofibrosarcoma, leiomyosarcoma, MPNST and fibrosarcoma) based on differentially expressed genes and evaluated similarities. Three of the 21 MFH samples showed marked similarities to one of the five sarcoma types, which were supported by histological findings. Although most of the remaining 18 MFH samples showed little or no histological resemblance to one of the five sarcoma types, 12 of them showed moderate similarities in terms of gene expression. These results explain the heterogeneity of MFH and show that the majority of MFHs could be reclassified into pleomorphic subtypes of other sarcomas. Taken together, gene expression profiling could be a useful tool to unveil the difference in the underlying molecular backgrounds, which leads to a rational taxonomy and diagnosis of a diverse group of soft tissue sarcomas. PMID- 17464316 TI - Expression of Sox2 in mature and immature teratomas of central nervous system. AB - Sox2 is a transcription factor that plays a critical role in the maintenance of the self-renewal capability of neural stem cells. This study was undertaken to investigate the expression pattern of Sox2 in mature and immature teratomas of the central nervous system. Sox2 immunohistochemistry was performed in 14 cases of central nervous system teratoma: five mature, five immature teratomas, and four mixed germ cell tumors with a prominent teratoma component. Fetal brain tissue was used as a normal control. Immunofluorescence with double labeling of stem cells and neuroglial cell markers was used for phenotyping of Sox2-positive cells. In all cases of immature teratomas, positive reactivity to Sox2 was observed in primitive neuroepithelial tissues. Sox2 was not expressed in mature tissues, except in some cuboidal or columnar epithelium of endodermal origin. In mature teratomas, Sox2 expression was limited to some endodermal epithelium in two cases, and no Sox2 expression was observed in the other three cases. The majority of Sox2-positive neuroepithelial cells also expressed neural stem cell markers, nestin and vimentin. Sox2 and neuronal and oligodendroglial markers were expressed in a mutually exclusive manner. However, mature astroglial cells coexpressed Sox2 and GFAP. In fetal brain, Sox2 was mainly expressed in ventricular and subventricular zones. Since Sox2 is strongly expressed in the primitive neuroepithelial tissues of central nervous system immature teratomas, it may be a useful biomarker for the diagnosis and quantitative grading of central nervous system immature teratomas. PMID- 17464317 TI - Expression of MYCN in pediatric synovial sarcoma. AB - Synovial sarcoma accounts for between 6 and 10% of childhood sarcomas and histological diagnosis can be challenging, even for experienced pathologists. Several other tumors enter the differential diagnosis, including malignant peripheral nerve sheath tumor, Ewing sarcoma/primitive neuroectodermal tumor and undifferentiated sarcoma. Several recent reports utilizing expression array techniques have documented expression of the MYCN oncogene in synovial sarcoma. In order to more fully investigate this finding, a series of 12 synovial sarcomas and 29 other sarcomas (four malignant peripheral nerve sheath tumors, 15 Ewing sarcoma/primitive neuroectodermal tumors, 10 undifferentiated sarcomas) were examined for MYCN expression and gene amplification. By RT-PCR, nine of 12 synovial sarcomas (75%) expressed MYCN. Five synovial sarcomas (42%) expressed MYCN at high levels. Of the other sarcomas, one malignant peripheral nerve sheath tumor (25%) and five Ewing sarcoma/primitive neuroectodermal tumors (33%) expressed MYCN at low levels, and all other cases were negative for MYCN. None of the synovial sarcomas had genomic amplification, suggesting that high MYCN expression levels resulted from epigenetic phenomena. Examination of selected downstream targets of MYCN in synovial sarcoma revealed expression of MCM7 (minichromosome maintenance protein 7) in all synovial sarcomas, and expression of nestin (n=10; 83%), ID2 (inhibitor of DNA binding protein 2) (n=6; 50%) and MRP1 (multidrug resistance protein 1) (n=1; 8%) in a subset of synovial sarcomas. Expression of downstream targets did not correlate with expression of MYCN. Neither MYCN nor expression of downstream targets significantly correlated with metastases at presentation, progression-free survival or overall survival in this small series. In summary, high levels of MYCN expression was useful for distinguishing synovial sarcoma from other childhood-spindled cell sarcomas with specificity and sensitivity of 100 and 42%, respectively, in this series. The clinical and biological significance of this finding deserves further study. PMID- 17464318 TI - Colorectal adenocarcinoma with micropapillary pattern and its association with lymph node metastasis. AB - Micropapillary carcinoma has been reported as an aggressive variant of carcinoma in several organs, where it is associated with frequent lymphovascular invasion and poor clinical outcome. This study explored the clinicopathological features of colorectal adenocarcinoma with a micropapillary carcinoma component and compared them with those of conventional colorectal adenocarcinoma. One hundred seventy-eight consecutive cases of surgically resected colorectal carcinomas were studied for tumor size, type, depth of invasion, nodal and distant metastases, tumor stage, and percentage and extent of micropapillary component. Among 178 cases of colorectal carcinoma, 34 (19.1%) cases had a micropapillary component, which ranged from 5 to 60% of the entire tumor. Lymph node metastasis was identified in 25 of 34 (73.5%) carcinomas with micropapillary component, whereas they were detected in 61 of 144 (42.4%) cases without micropapillary component (P=0.001). Lymphovascular invasion was identified more frequently in carcinoma with micropapillary component (41.2%) than carcinoma without micropapillary component (20.1%; P<0.05). Distant metastases occurred in 4 of 34 cases (11.7%) with micropapillary component and in 10 of 144 cases (6.9%) without micropapillary component (P=0.311). Multivariate regression analysis demonstrated that the presence of micropapillary component, as well as tumor stage and lymphovascular invasion are independent predictors of regional nodal metastasis. PMID- 17464319 TI - Lateral peritumoral lymphatic vessel invasion can predict lymph node metastasis in esophageal squamous cell carcinoma. AB - Lymph node metastasis is an important prognostic factor in many types of cancer. We investigated the clinical significance of lymphangiogenesis and lymphatic vessel invasion in esophageal squamous cell carcinoma. We evaluated lymphatic vessel density and lymphatic vessel invasion in the intratumoral, peritumoral and normal compartments using D2-40 immunostaining. In addition, the peritumoral compartment was divided into the lateral peritumoral compartment and the non lateral peritumoral compartment. The lymphatic vessel density was higher in the peritumoral and intratumoral compartments than in the normal compartment. However, the lymphatic vessel density did not correlate with any pathological parameters including lymph node metastasis. Intratumoral and peritumoral lymph vessels were small and collapsed while normal lymphatic vessels and lymphatic vessels with lymphatic vessel invasion were dilated and large. The presence of lymphatic vessel invasion, in the lateral peritumoral compartment but nowhere else, significantly correlated with lymph node metastasis. These results suggest that lymphangiogenesis might occur with esophageal cancer, but it does not play a direct role in lymphatic vessel invasion and lymph node metastasis. Peritumoral lymphatic vessel invasion, especially in the lateral peritumoral compartment, should imply a high probability of regional lymph node metastasis. PMID- 17464320 TI - Prognostic significance of NPM-ALK fusion transcript overexpression in ALK positive anaplastic large-cell lymphoma. AB - In anaplastic large-cell lymphomas positive for anaplastic lymphoma kinase (ALK) protein, the ALK gene is most commonly fused to the NPM gene, and less commonly to TPM3, TFG, ATIC, and other rare genes. Although this lymphoma is generally associated with a favorable clinical outcome, 25% of the patients die of the disease within 5 years. In this study, we developed three assays, all of which can be used with archival formalin-fixed, paraffin-embedded tissues: (1) a sensitive reverse transcription-polymerase chain reaction (RT-PCR) assay for various X-ALK fusion genes, (2) a 5' rapid amplification of cDNA ends (RACE) assay to identify unknown fusion partners, and (3) a real-time RT-PCR assay to quantify the amount of the NPM-ALK fusion transcript. In 26 cases of ALK(+) anaplastic large-cell lymphoma, the RT-PCR assay showed that the ALK was fused to NPM in 21 cases, to TPM3 in three, and to TFG in one. The 5' RACE assay detected ATIC-ALK fusion in the remaining case. The real-time quantitative RT-PCR assay showed that the NPM-ALK transcript was over expressed in four of 20 quantifiable cases. Patients with NPM-ALK overexpression showed a significantly unfavorable overall survival compared with those with a low expression of this transcript. The RT-PCR and 5' RACE assays developed here may be useful for identification of known and unknown gene partners fused to the ALK gene. Overexpression of the NPM ALK fusion transcript may be associated with a poor prognosis of the patients with ALK(+) anaplastic large-cell lymphomas. PMID- 17464321 TI - Mitochondrial dysfunction in Parkinson's disease. PMID- 17464322 TI - Delayed neuronal death after brain trauma involves p53-dependent inhibition of NF kappaB transcriptional activity. AB - Acute and chronic neurodegeneration, for example, following brain injury or Alzheimer's disease, is characterized by programmed death of neuronal cells. The present study addresses the role and interaction of p53- and NF-kappaB-dependent mechanisms in delayed neurodegeneration following traumatic brain injury (TBI). After experimental TBI in mice p53 rapidly accumulated in the injured brain tissue and translocated to the nucleus of damaged neurons, whereas NF-kappaB transcriptional activity simultaneously declined. Post-traumatic neurodegeneration correlated with the increase in p53 levels and was significantly reduced by the selective p53 inhibitor pifithrin-alpha (PFT). Strikingly, this protective effect was observed even when PFT treatment was delayed up to 6 h after trauma. Inhibition of p53 activity resulted in the concomitant increase in NF-kappaB transcriptional activity and upregulation of NF kappaB-target proteins, for example X-chromosomal-linked inhibitor of apoptosis (XIAP). It is interesting to note that inhibition of XIAP abolished the neuroprotective effects of PFT in cultured neurons exposed to camptothecin, glutamate, or oxygen glucose deprivation. In conclusion, delayed neuronal cell death after brain trauma is mediated by p53-dependent mechanisms that involve inhibition of NF-kappaB transcriptional activity. Hence, p53 inhibition provides a promising approach for the treatment of acute brain injury, since it blocks apoptotic pathways and concomitantly triggers survival signaling with a therapeutic window relevant for clinical applications. PMID- 17464323 TI - The cell cycle inhibitor p57(Kip2) promotes cell death via the mitochondrial apoptotic pathway. AB - The p57(Kip2) gene belongs to the Cip/Kip family of cyclin-dependent kinase (CDK) inhibitors and has been suggested to be a tumor suppressor gene, being inactivated in various types of human cancers. However, little is known concerning p57(Kip2) possible interplay with the apoptotic cell death machinery and its possible implication for cancer. Here, we report that selective p57(Kip2) expression sensitizes cancer cells to apoptotic agents such as cisplatin, etoposide and staurosporine (STS) via a mechanism, which does not require p57(Kip2)-mediated inhibition of CDK. Translocation of p57(Kip2) to mitochondria occurs within 20 min after STS application. In fact, p57(Kip2) primarily promotes the intrinsic apoptotic pathways, favoring Bax activation and loss of mitochondrial transmembrane potential, consequent release of cytochrome-c into cytosol, caspase-9 and caspase-3 activation. In accordance, Bcl2 overexpression or voltage-dependent anion channel (VDAC) inhibition is able to inhibit p57(Kip2) cell death promoting effect. Thus, in addition to its established function in control of proliferation, these results reveal a mechanism whereby p57(Kip2) influences the mitochondrial apoptotic cell death pathway in cancer cells. PMID- 17464324 TI - A new role for NF-kappaB in angiogenesis inhibition. PMID- 17464325 TI - The Drosophila caspases Strica and Dronc function redundantly in programmed cell death during oogenesis. AB - Programmed cell death (PCD) in the Drosophila ovary occurs either during mid oogenesis, resulting in degeneration of the entire egg chamber or during late oogenesis, to facilitate the development of the oocyte. PCD during oogenesis is regulated by mechanisms different from those that control cell death in other Drosophila tissues. We have analyzed the role of caspases in PCD of the female germline by examining caspase mutants and overexpressing caspase inhibitors. Imprecise P-element excision was used to generate mutants of the initiator caspase strica. While null mutants of strica or another initiator caspase, dronc, display no ovary phenotype, we find that strica exhibits redundancy with dronc, during both mid- and late oogenesis. Ovaries of double mutants contain defective mid-stage egg chambers similar to those reported previously in dcp-1 mutants, and mature egg chambers with persisting nurse cell nuclei. In addition, the effector caspases drice and dcp-1 also display redundant functions during late oogenesis, resulting in persisting nurse cell nuclei. These findings indicate that caspases are required for nurse cell death during mid-oogenesis, and participate in developmental nurse cell death during late oogenesis. This reveals a novel pathway of cell death in the ovary that utilizes strica, dronc, dcp-1 and drice, and importantly illustrates strong redundancy among the caspases. PMID- 17464326 TI - Atypical, bidirectional regulation of cadmium-induced apoptosis via distinct signaling of unfolded protein response. AB - Cadmium is a widely distributed nephrotoxic metal that causes renal tubular injury. In this report, we investigated involvement of endoplasmic reticulum (ER) stress and individual unfolded protein responses in cadmium-initiated apoptosis of tubular epithelial cells. Cadmium chloride (CdCl(2)) induced expression of endogenous ER stress markers, GRP78, GRP94 and CHOP in vitro and in vivo, and subsequently caused cytological changes typical of apoptosis. Attenuation of ER stress by transfection with ER chaperone GRP78 or ORP150 suppressed CdCl(2) triggered apoptosis. In response to CdCl(2), phosphorylation of RNA-dependent protein kinase-like ER kinase (PERK) and eukaryotic translation initiation factor 2alpha (eIF2alpha) was observed. Enhanced phosphorylation of eIF2alpha attenuated, whereas inhibition of eIF2alpha exacerbated CdCl(2)-induced apoptosis. Activating transcription factor 6 (ATF6) was also activated by CdCl(2) and blockade of this process suppressed induction of CHOP and thereby improved cell survival. CdCl(2) also triggered activation of the inositol-requiring ER-to nucleus signal kinase 1 (IRE1)-X-box-binding protein 1 (XBP1) pathway and inhibition of XBP1 attenuated apoptosis independent of GRP78 and CHOP. c-Jun N terminal kinase (JNK), another molecule downstream of IRE1, was also phosphorylated by CdCl(2) and its inhibition attenuated apoptosis. These results evidenced bidirectional regulation of apoptosis in cadmium-exposed cells. The ATF6 and IRE1 pathways cooperatively caused apoptosis via induction of CHOP, activation of XBP1 and phosphorylation of JNK, and the PERK-eIF2alpha pathway counteracted the proapoptotic processes. PMID- 17464327 TI - Aberrant quality control in the endoplasmic reticulum impairs the biosynthesis of pulmonary surfactant in mice expressing mutant BiP. AB - Accumulation of misfolded proteins in the endoplasmic reticulum (ER) induces the unfolded protein response (UPR), which alleviates protein overload in the secretory pathway. Although the UPR is activated under diverse pathological conditions, its physiological role during development and in adulthood has not been fully elucidated. Binding immunoglobulin protein (BiP) is an ER chaperone, which is central to ER function. We produced knock-in mice expressing a mutant BiP lacking the retrieval sequence to cause a defect in ER function without completely eliminating BiP. In embryonic fibroblasts, the UPR compensated for mutation of BiP. However, neonates expressing mutant BiP suffered respiratory failure due to impaired secretion of pulmonary surfactant by alveolar type II epithelial cells. Expression of surfactant protein (SP)-C was reduced and the lamellar body was malformed, indicating that BiP plays a critical role in the biosynthesis of pulmonary surfactant. Because pulmonary surfactant requires extensive post-translational processing in the secretory pathway, these findings suggest that in secretory cells, such as alveolar type II cells, the UPR is essential for managing the normal physiological ER protein overload that occurs during development. Moreover, failure of this adaptive mechanism may increase pulmonary susceptibility to environmental insults, such as hypoxia and ischemia, ultimately leading to neonatal respiratory failure. PMID- 17464328 TI - A cut short to death: Parl and Opa1 in the regulation of mitochondrial morphology and apoptosis. AB - Mitochondria are crucial amplifiers of death signals. They release cytochrome c and other pro-apoptotic factors required to fully activate effector caspases. This release is accompanied by fragmentation of the mitochondrial reticulum and by remodelling of the internal structure of the organelle. Here we review data supporting the existence of a regulatory network in the inner mitochondrial membrane that includes optic atrophy 1 (Opa1), a dynamin-related protein, and presenilin-associated rhomboid-like (Parl), a rhomboid protease. Opa1 regulates remodelling of the cristae independent of its effect on fusion. Cristae remodelling conversely requires Parl, which participates in the production of a soluble form of Opa1 retrieved together with the integral membrane one in oligomers that are disrupted early during apoptosis. Parl itself is regulated by proteolysis to generate a cleaved form, which in turn modulates the shape of the mitochondrial reticulum. Cleavage of Parl depends on its phosphorylation state around the cleavage site, implicating mitochondrial kinases and phosphatases in the regulation of mitochondrial shape. PMID- 17464329 TI - Novel role of HDAC inhibitors in AML1/ETO AML cells: activation of apoptosis and phagocytosis through induction of annexin A1. AB - The chimeric fusion protein AML1-ETO, created by the t(8;21) translocation, recruits histone deacetylase (HDAC) to AML1-dependent promoters, resulting in transcriptional repression of the target genes. We analyzed the transcriptional changes in t(8;21) Kasumi-1 AML cells in response to the HDAC inhibitors, depsipeptide (FK228) and suberoylanilide hydroxamic acid (SAHA), which induced marked growth inhibition and apoptosis. Using cDNA array, annexin A1 (ANXA1) was identified as one of the FK228-induced genes. Induction of ANXA1 mRNA was associated with histone acetylation in ANXA1 promoter and reversal of the HDAC dependent suppression of C/EBPalpha by AML1-ETO with direct recruitment of C/EBPalpha to ANXA1 promoter. This led to increase in the N-terminal cleaved isoform of ANXA1 protein and accumulation of ANXA1 on cell membrane. Neutralization with anti-ANXA1 antibody or gene silencing with ANXA1 siRNA inhibited FK228-induced apoptosis, suggesting that the upregulation of endogenous ANXA1 promotes cell death. FK228-induced ANXA1 expression was associated with massive increase in cell attachment and engulfment of Kasumi-1 cells by human THP 1-derived macrophages, which was completely abrogated with ANXA1 knockdown via siRNA transfection or ANXA1 neutralization. These findings identify a novel mechanism of action of HDAC inhibitors, which induce the expression and externalization of ANXA1 in leukemic cells, which in turn mediates the phagocytic clearance of apoptotic cells by macrophages. PMID- 17464330 TI - Mouse embryonic stem cells are hypersensitive to apoptosis triggered by the DNA damage O(6)-methylguanine due to high E2F1 regulated mismatch repair. AB - Exposure of stem cells to genotoxins may lead to embryonic lethality or teratogenic effects. This can be prevented by efficient DNA repair or by eliminating genetically damaged cells. Using undifferentiated mouse embryonic stem (ES) cells as a pluripotent model system, we compared ES cells with differentiated cells, with regard to apoptosis induction by alkylating agents forming the highly mutagenic and killing DNA adduct O(6)-methylguanine. Upon treatment with N-methyl-N'-nitro-N-nitrosoguanidine (MNNG), ES cells undergo apoptosis at much higher frequency than differentiated cells, although they express a high level of the repair protein O(6)-methylguanine-DNA methyltransferase (MGMT). Apoptosis induced by MNNG is due to O(6)-methylguanine DNA adducts, since inhibition of MGMT sensitized ES cells. The high sensitivity of ES cells to O(6)-methylating agents is due to high expression of the mismatch repair proteins MSH2 and MSH6 (MutSalpha), which declines during differentiation. High MutSalpha expression in ES cells was related to a high hyperphosphorylated retinoblastoma (ppRb) level and E2F1 activity that upregulates MSH2, causing, in turn, stabilization of MSH6. Non-repaired O(6)-methylguanine adducts were shown to cause DNA double-stranded breaks, stabilization of p53 and upregulation of Fas/CD95/Apo-1 at significantly higher level in ES cells than in fibroblasts. The high apoptotic response of ES cells to O(6)-methylguanine adducts may contribute to reduction of the mutational load in the progenitor population. PMID- 17464331 TI - JDP2 suppresses adipocyte differentiation by regulating histone acetylation. AB - Among the events that control cellular differentiation, the acetylation of histones plays a critical role in the regulation of transcription and the modification of chromatin. Jun dimerization protein 2 (JDP2), a member of the AP 1 family, is an inhibitor of such acetylation and contributes to the maintenance of chromatin structure. In an examination of Jdp2 'knock-out' (KO) mice, we observed elevated numbers of white adipocytes and significant accumulation of lipid in the adipose tissue in sections of scapulae. In addition, mouse embryo fibroblasts (MEFs) from Jdp2 KO mice were more susceptible to adipocyte differentiation in response to hormonal induction and members of the CCAAT/enhancer-binding proteins (C/EBP) gene family were expressed at levels higher than MEFs from wild-type mice. Furthermore, JDP2 inhibited both the acetylation of histone H3 in the promoter of the gene for C/EBPdelta and transcription from this promoter. Our data indicate that JDP2 plays a key role as a repressor of adipocyte differentiation by regulating the expression of the gene for C/EBPdelta via inhibition of histone acetylation. PMID- 17464332 TI - Siva is an apoptosis-selective p53 target gene important for neuronal cell death. AB - p53 plays a central role in neuronal cell death resulting from acute injury or disease. To define the pathway by which p53 triggers apoptosis, we used microarray analysis to identify p53 target genes specifically upregulated during apoptosis but not cell cycle arrest. This analysis identified a small subset of targets highly selective for the p53 apoptotic response, including Siva, a proapoptotic protein whose function is not well understood. Siva's expression pattern suggests that it plays an instructive role in apoptosis, and accordingly, we demonstrate that Siva is essential for p53-dependent apoptosis in cerebellar granule neurons. In addition, we determine that endogenous Siva is associated with the plasma membrane and that Caspase-8 and Bid are important for neuronal apoptosis. Our studies highlight the participation of membrane signaling events in p53's apoptotic program in primary neurons and have significant implications for understanding the mechanisms underlying pathogenesis after neuronal injury and in neurodegenerative diseases. PMID- 17464334 TI - Expectations for specialized knowledge in oncology: time to reassess. PMID- 17464333 TI - Tumour necrosis factor-alpha inhibits adipogenesis via a beta catenin/TCF4(TCF7L2)-dependent pathway. AB - Tumour necrosis factor-alpha (TNF-alpha), a proinflammatory cytokine, is a potent negative regulator of adipocyte differentiation. However, the mechanism of TNF alpha-mediated antiadipogenesis remains incompletely understood. In this study, we first confirm that TNF-alpha inhibits adipogenesis of 3T3-L1 preadipocytes by preventing the early induction of the adipogenic transcription factors peroxisome proliferator-activated receptor-gamma (PPARgamma) and CCAAT/enhancer binding protein-alpha (C/EBPalpha). This suppression coincides with enhanced expression of several reported mediators of antiadipogenesis that are also targets of the Wnt/beta-catenin/T-cell factor 4 (TCF4) pathway. Indeed, we found that TNF-alpha enhanced TCF4-dependent transcriptional activity during early antiadipogenesis, and promoted the stabilisation of beta-catenin throughout antiadipogenesis. We analysed the effect of TNF-alpha on adipogenesis in 3T3-L1 cells in which beta catenin/TCF signalling was impaired, either via stable knockdown of beta-catenin, or by overexpression of dominant-negative TCF4 (dnTCF4). The knockdown of beta catenin enhanced the adipogenic potential of 3T3-L1 preadipocytes and attenuated TNF-alpha-induced antiadipogenesis. However, beta-catenin knockdown also promoted TNF-alpha-induced apoptosis in these cells. In contrast, overexpression of dnTCF4 prevented TNF-alpha-induced antiadipogenesis but showed no apparent effect on cell survival. Finally, we show that TNF-alpha-induced antiadipogenesis and stabilisation of beta-catenin requires a functional death domain of TNF-alpha receptor 1 (TNFR1). Taken together these data suggest that TNFR1-mediated death domain signals can inhibit adipogenesis via a beta-catenin/TCF4-dependent pathway. PMID- 17464335 TI - A remedy for biomarker addiction: back to rational anticancer drug development. PMID- 17464336 TI - Chemoradiotherapy for localized esophageal cancer: regimen selection and molecular mechanisms of radiosensitization. AB - Concurrent chemoradiotherapy administered either before surgery or as definitive treatment has a central role in the multimodality treatment of locally advanced esophageal cancer. Initial studies of this combined-modality regimen were based on models of squamous-cell cancers from other primary sites; this approach progressed from use of bleomycin or fluorouracil plus cisplatin concurrent with radiation in early trials, to the integration of taxanes, camptothecins and platinum analogs in recent trials. These trials demonstrated the tumoricidal effect of concurrent chemotherapy and radiotherapy and showed the survival advantages of this approach. Preoperative concurrent chemoradiation is used to downstage the tumor, ideally to a pathological complete response status in which there is no residual tumor in the resected primary and nodal tissues. A pathological complete response is associated with long-term survival but occurs in a minority (30%) of patients. While clinical trials have demonstrated an improvement in survival with concurrent chemoradiotherapy this effect is limited, as indicated by the plateau in survival beyond 5 years of approximately 30% or less. The recent clinical development of biologic, targeted therapies provides a new avenue for the study of chemoradiotherapy and an opportunity to increase long term survival. PMID- 17464337 TI - Medulloblastoma: tumorigenesis, current clinical paradigm, and efforts to improve risk stratification. AB - Medulloblastoma is the most common brain malignancy in children and tremendous advances have recently been made in understanding the pathogenesis of this tumor. The Hedgehog and Wingless signaling pathways are implicated in medulloblastoma development, and both pathways were discovered as a result of analyses of genetic syndromes associated with the tumor. Over the past 80 years, considerable progress has been made in the treatment of what was once a fatal disease. The first survival reports followed the introduction of craniospinal irradiation, and yet the success of this modality, which continues to be a central component of treatment regimens for patients older than 3 years, comes at a significant cost. The present challenge in medulloblastoma treatment is to improve upon existing survival rates and to minimize the side effects of treatment. The current tools of clinical risk assessment fail to adequately identify patients older than 3 years who require less radiation and those who require more radiation. Significant effort has been made to improve clinical risk stratification and titration of treatment by analyzing properties of the tumor cells themselves for prognostic significance. These efforts include identifying histopathologic, cytogenetic, and molecular features that may correlate with prognosis. PMID- 17464338 TI - Mechanisms of disease: methyl-binding domain proteins as potential therapeutic targets in cancer. AB - The methyl-CpG-binding domain (MBD) proteins 'read' and interpret the methylation moieties on DNA, and thus are critical mediators of many epigenetic processes. Currently, the MBD family comprises five members; MBD1, MBD2, MBD3, MBD4 and MeCP2. Although not a 'classical' MBD protein, Kaiso also mediates transcriptional repression by using zinc finger domains to bind its targets. Since DNA hypermethylation is a well-recognized mechanism underlying gene silencing events in both tumorigenesis and drug resistance, it is likely that the MBD proteins may be important modulators of tumorigenesis. We review the recent work addressing this possibility, and discuss several of the MBD proteins as potentially excellent novel therapeutic targets. PMID- 17464340 TI - Effects of estradiol and 4-hydroxytamoxifen on the conformation, thermal stability, and DNA recognition of estrogen receptor beta. AB - Estrogen receptors (ERalpha and ERbeta) are ligand-activated transcription factors. We examined the effects of estradiol (E2), 4-hydroxytamoxifen (HT), and the estrogen response element (ERE) on the helical content and thermal unfolding of ERbeta. A circular dichroism (CD) spectrum of ERbeta showed changes at 210 and 222 nm that were due to the presence of E2, which is indicative of partial unfolding. In contrast, HT did not alter the CD spectrum of ERbeta. The addition of E2 + ERE caused an increase in the alpha-helical content and an increase in the temperature midpoint of folding transition (TM) from 39 +/- 0.7 degrees C to 57.2 +/- 1 degrees C. The addition of E2 + mutant ERE, or E2 + control oligonucleotide, increased the TM of ERbeta to 45 +/- 2 degrees C only. In the presence of HT, ERbeta yielded similar TM values (55-58 degrees C) with ERE, mutant ERE, or control oligodeoxynucleotide. The binding affinity of ERbeta for ERE increased 125.7-fold as a result of the presence of E2, but only 4-fold as a result of HT. These results demonstrate coupled effects of E2 and ERE on ERbeta stability and binding affinity. The increased thermal stability of HT-ERbeta-ERE was associated with reduced specificity of ERbeta-ERE recognition, illustrating profound differences in conformational states of ERbeta induced by E2 and HT. PMID- 17464339 TI - Complete pathological response to bevacizumab and chemoradiation in advanced rectal cancer. AB - BACKGROUND: Localized rectal cancer responds well to 5-fluorouracil and radiation based regimens. A phase I-II trial is currently testing the efficacy of adding bevacizumab, a VEGF-specific antibody, to standard chemoradiotherapy. The case presented here is a complete pathological response seen in a patient with extensive and locally invasive carcinoma after receiving this combined treatment. INVESTIGATIONS: Physical examination, rectal ultrasound, PET-CT scan, laboratory tests, proctoscopic examination, chest radiograph, rectal forcep biopsies with immunohistochemistry, and protein and flow cytometric analyses. DIAGNOSIS: Large, invasive, ultrasound stage T4 carcinoma of the rectum, which was positive for survivin. MANAGEMENT: One 2-week cycle of bevacizumab alone, followed by 3 cycles of bevacizumab with continuous 5-fluorouracil infusion, and external-beam radiation therapy given 5 days per week to the pelvis, abdominoperineal resection with posterior vaginectomy, hysterectomy and bilateral salpingo-oophorectomy. PMID- 17464341 TI - The ASP receptor C5L2 is regulated by metabolic hormones associated with insulin resistance. AB - Acylation-stimulating protein (ASP) and interaction with its receptor C5L2 influences adipocyte metabolism. We examined insulin resistance and differentiation-mediated regulation of C5L2 and the mechanistic impact on both C5L2 cell-surface protein and ligand binding to the receptor. C5L2 mRNA increased 8.7-fold with differentiation in 3T3-L1 cells (p < 0.0001) by day 9. In preadipocytes, insulin and dexamethasone increased C5L2 mRNA (1 micromol/L insulin resulted in a 2.6-fold increase, p < 0.01; 10 nmol/L dexamethasone resulted in a 17.9-fold increase, p < 0.01) and C5L2 cell-surface protein (100 nmol insulin resulted in a 2.7-fold increase, p < 0.001; 10 nmol/L dexamethasone resulted in a 2.8-fold increase, p < 0.001). In adipocytes, 100 nmol/L insulin increased C5L2 mRNA and ASP binding (respectively, 1.3-fold, p < 0.01; and 2.4 fold, p < 0.05). Dexamethasone decreased ligand binding (-60%, p < 0.02) without changing mRNA. Tumor necrosis factor alpha decreased C5L2 mRNA (-88% in preadipocytes and -38% in adipocytes, p < 0.001), C5L2 cell-surface protein (-53% in preadipocytes, p < 0.0001), and ASP binding (-60% and -49% in, respectively, preadipocytes and adipocytes, p < 0.05). Conversely, 1 micromol/L and 10 nmol/L rosiglitazone increased, respectively, C5L2 mRNA (9.3-fold, p < 0.0001) and ASP binding (2.4-fold, p < 0.05). Thus, C5L2 mRNA increases with differentiation, insulin, and thiazolidinedione treatment, and decreases with tumor necrosis factor alpha, all of which results in functional changes in ASP-C5L2 response and may have implications for human metabolism. PMID- 17464342 TI - A novel Amoeba proteus 120 kDa actin-binding protein with only 1 filamin repeat and a coiled-coil region. AB - A novel 120 kDa actin-binding protein (ApABP-F1) was found in Amoeba proteus. It was distributed throughout the cytoplasm, mainly in the subplasma membrane and perinuclear-nuclear areas, enriched in actin. The full-length cDNA of ApABP consisted of 2672 nucleotides with an open reading frame of 878 amino acids, giving a ~95 kDa protein with a theoretical pI value of 5.11. It had a novel domain organization pattern: the N terminus (residues 1-104) contained 1 calponin homology (CH) domain, followed by only 1 region that was homologous to the filamin repeat (FR, residues 209-324), and a central region (residues 344-577) exhibiting a very high probability of coiled-coil formation, probably engaged in the observed protein dimerization. A phylogenetic tree constructed for CH domains from 25 various proteins revealed that the CH domain of ApABP was most related to that of the hypothetical mouse KIAA0903-like protein, whereas not much relationship to either filamins or the gelation factor (ABP-120) of Dictyostelium discoideum and Entamoeba histolytica was found. PMID- 17464343 TI - Differential effects of c-Ras upon transformation, adipocytic differentiation, and apoptosis mediated by the simian virus 40 large tumor antigen. AB - To investigate the functional relationship between the ability of the simian virus 40 large tumor antigen (TAg) to transform and its ability to block adipocytic differentiation and induce apoptosis, we expressed TAg in C3H10T1/2 (10T1/2)-derived preadipocytes. The results demonstrated that differentiation could be suppressed at lower TAg levels than at the levels required for full neoplastic conversion. Progressively higher TAg levels were accompanied by apoptosis induction in this system. To further examine the role of the cellular Ras protooncogene product (Ras) in TAg function, TAg was expressed in 10T1/2 derived preadipocytes rendered deficient in Ras activity by transfection with inducible or constitutive antisense ras gene constructs. The results indicated that Ras is required for TAg-mediated transformation and for suppression of adipocytic differentiation, while TAg-mediated apoptosis following serum starvation was independent from Ras action. Unexpectedly, our results further demonstrated a dramatic reduction in the levels of the TAg protein itself as differentiation progressed in Ras-knockdown cells, with a concomitant reduction in TAg's ability to induce apoptosis as a result. These findings suggest that Ras, although cytoplasmic, is an integral component of the pathway whereby TAg, an oncoprotein believed to have primarily nuclear targets, suppresses differentiation or induces neoplastic conversion of murine preadipocytes. PMID- 17464344 TI - Mitochondria-independent morphological and biochemical apoptotic alterations promoted by the anti-tumor agent bleomycin in Saccharomyces cerevisiae. AB - Bleomycin is a highly potent cytotoxic and genotoxic agent used in the chemotherapy of various types of tumors. It is a radiomimetic anticancer drug that produces single- and double-stranded DNA breaks in a catalytic way. Using Saccharomyces cerevisiae as a model system, we show that when a high amount of bleomycin molecules is internalized (100 micromol/L), morphological changes identical to those usually associated with apoptosis, i.e., a sub-G1 region peak, chromatin condensation, and very rapid DNA fragmentation into oligonucleosomal sized fragments, are observed. The known bleomycin inhibitors cobalt and EDTA were able to prevent bleomycin nucleasic activity and thus apoptotic cell death. However, both oligomycin, a potent inhibitor of the mitochondrial F0F1-ATPase, and antimycin, a drug affecting mitochondria respiration, were unable to prevent the bleomycin-induced apoptotic-like cell death. These results suggest that high bleomycin concentrations induce an apoptosis-like mitochondria-independent cell death in yeast. PMID- 17464345 TI - Novel irreversible caspase-1 inhibitor attenuates the maturation of intracellular interleukin-1beta. AB - Caspase-1, the most efficient enzyme in processing the proinflammatory cytokines interleukin 1beta and interleukin 18 in humans, is associated with inflammatory diseases such as rheumatoid arthritis, osteoarthritis, and some neuronal diseases. We previously reported that isoquinoline-1,3,4-trione and its derivatives are novel caspase-3 inhibitors that could attenuate apoptosis in vitro and in vivo. Here we report a novel derivative of isoquinoline-1,3,4-trione that is highly potent in inhibiting caspase-1 activity in an irreversible and slow-binding manner, thus inhibiting cellular caspase-1 activity and the maturation of interleukin 1beta in U-937 cells. PMID- 17464346 TI - The inhibitory effect of simvastatin on the ADMA-induced inflammatory reaction is mediated by MAPK pathways in endothelial cells. AB - Asymmetric dimethylarginine (ADMA), an endogenous inhibitor of nitric oxide synthase, is emerging as a key contributor for endothelial dysfunction associated with inflammation. Statins can inhibit vascular inflammatory reaction and improve endothelial function. The aim of this study was to investigate in human endothelial cells the signaling pathways of ADMA-induced inflammatory reaction and potential inhibitory effects of simvastatin. Endothelial cells were cultured and used for all of the studies. Tumor necrosis factor-alpha(TNF-alpha) and soluble intercellular adhesion molecule-1 (sICAM-1) were determined by enzyme linked immunosorbent assay. Nuclear factor-kappaB (NF-kappaB) was assayed by electrophoretic mobility shift assay. The activation of mitogen-activated protein kinases (MAPKs), including p38 MAPK and extracellular signal-related kinase (ERK(1/2)), were characterized by Western blot analysis. Treatment with ADMA (3 30 micromol/L) increased the concentration of sICAM-1 in a dose-dependent manner. ADMA (30 micromol/L) significantly enhanced the concentrations of TNF-alpha and sICAM-1, the activity of NF-kappaB and the phosphorylation of p38 MAPK and ERK(1/2). The increased secretion of TNF-alpha and sICAM-1 and the increased activity of NF-kappaB by ADMA were altered by SB203580 (5 micromol/L) or PD98059 (20 micromol/L), but not by LY294002 (20 micromol/L). Simvastatin (0.1, 0.5, or 2.5 micromol/L) markedly inhibited the elevated concentrations of TNF-alpha and sICAM-1, the activity of NF-kappaB, and the phosphorylation of p38 MAPK and ERK(1/2) induced by ADMA. Simvastatin inhibited ADMA-induced inflammatory reaction by p38 MAPK and ERK(1/2) pathways in cultured endothelial cells. PMID- 17464347 TI - Effect of cAMP-dependent protein kinase A (PKA) on HCV nucleocapsid assembly and degradation. AB - The primary function of the hepatitis C virus (HCV) core protein is genome encapsidation. Core protein is also subject to post-translational modifications that can impact on the assembly process. In this report, we have studied the effect of cAMP-dependent protein kinase A (PKA) phosphorylation on its assembly and stability in a yeast Pichia pastoris expression system. We have recently shown that co-expression of the human signal peptide peptidase and core protein (amino acids 1-191) in yeast leads to the formation of nucleocapsid-like particles (NLPs) that are morphologically similar to the wild-type HCV capsid. In this system, we expressed mutants S53A and S116A and mutants S53D and S116D to abolish or mimic PKA phosphorylation, respectively. None of these mutations affected HCV assembly, but S116D led to the degradation of core protein. We also showed that nonenveloped NLPs were labelled in vitro by PKA, suggesting that the phosphorylation sites are available at the surface of the NLPs. The co-expression of human PKA with core and human signal peptide peptidase in yeast did not produce phosphorylated NLPs and led to a decreased accumulation of nonenveloped particles. Mutation S116A restored the core protein content. These results suggest that PKA phosphorylation can modulate HCV core levels in infected cells. PMID- 17464348 TI - Purification of 3 monomeric monocot mannose-binding lectins and their evaluation for antipoxviral activity: potential applications in multiple viral diseases caused by enveloped viruses. AB - Three monomeric monocot lectins from Zephyranthes carinata, Zephyranthes candida, and Gloriosa superba with carbohydrate specificity towards mannose derivatives and (or) oligomannose have been isolated and purified from their storage tissues. The lectins were purified by anion-exchange chromatography on DEAE-Sephacyl and by gel filtration chromatography on Biogel P-200 followed by high-performance liquid chromatography. The purified lectins, Z. carinata, Z. candida, and G. superba had molecular masses of 12, 11.5, and 12.5 kDa, respectively, as determined by gel filtration and SDS-PAGE, indicating that they are monomers. In a hapten inhibition assay, methyl-alpha-D-mannopyranoside inhibited agglutination of both Z. candida and Z. carinata; the latter was also inhibited by Man(alpha1 2)Man and Man(alpha1-3)Man. Gloriosa superba showed inhibition only with Man(alpha1-4)Man of all of the sugars and glycoproteins tested. All purified lectins agglutinated red blood cells from rabbit, whereas G. superba was also reactive towards erythrocytes from guinea pig. All of the lectins were nonglycosylated and did not require metal ions for their activity. They were labile above 60 degrees C and were affected by denaturing agents such as urea, thiourea, and guanidine-HCl. The lectins were virtually nonmitogenic, like other members of Amaryllidaceae and Liliaceae. Of the 3 lectins, G. superba was found to be highly toxic to the BSC-1 cell line (African green monkey kidney epithelial cells), while both of the Zephyranthes species showed significant in vitro inhibition of poxvirus replication in BSC-1 cells without any toxic effects to the cells. In addition, Z. candida also exhibited significant anticancer activity against SNB-78, a CNS human cancer cell line. PMID- 17464349 TI - Cloning and characterization of an alternative splicing transcript of the gene coding for human cytidine deaminase. AB - Human cytidine deaminase (HCD) catalyzes the deamination of cytidine or deoxycytidine to uridine or deoxyuridine, respectively. The genomic sequence of HCD is formed by 31 kb with 4 exons and several alternative splicing signals, but an alternative form of HCD has yet to be reported. Here we describe the cloning and characterization of a small form of HCD, HSCD, and it is likely to be a product of alternative splicing of HCD. The alignment of DNA sequences shows that the HSCD matches HCD in 2 parts, except for a deletion of 170 bp. Based on the HCD genome organization, exons 1 and 4 should be joined and all sequences of introns and exons 2 and 3 should be deleted by splicing. This alternative splicing shifted the translation of the reading frame from the point of splicing. The estimated molecular mass is 9.8 kDa, and this value was confirmed by Western blot and mass spectroscopy after expressing the gene fused with glutathionine-S transferase in the pGEX vector. The deletion and shift of the reading frame caused a loss of HCD activity, which was confirmed by enzyme assay and also with NIH3T3 cells modified to express HSCD and challenged against cytosine arabinoside. In this work we describe the identification and characterization of HSCD, which is the product of alternative splicing of the HCD gene. PMID- 17464350 TI - Inhibition of osteopontin would suppress angiogenesis in gastric cancer. AB - Osteopontin (OPN) plays an important role in tumorigenesis, tumor invasion, and metastasis in many types of cancers, including gastric cancer. Recently, much interest has been focused on the role of OPN in tumor angiogenesis. Our previous studies have shown that OPN is overexpressed, and associated with mean microvessel density in, the tissue samples of patients with gastric cancer. In the present study, we aimed to further determine and provide evidence for the role of OPN in gastric-cancer-associated angiogenesis by diminishing OPN expression in gastric cancer cells using the small interference RNA method, and then evaluate the effects of OPN on gastric cancer-associated angiogenesis by in vivo and in vitro assays. Our results revealed that reduced OPN production by gastric cancer cells would reduce the proliferation, migration, and tube formation of human umbilical vein endothelial cells, and lead to a lower microvessel density, i.e., angiogenesis, in transplanted tumors of mice. These data confirm the positive role of OPN in gastric-cancer-associated angiogenesis. PMID- 17464351 TI - Staurosporine inhibition of zipper-interacting protein kinase contractile effects in gastrointestinal smooth muscle. AB - Zipper-interacting protein kinase (ZIPK) is a serine-threonine kinase that has been implicated in Ca2+-independent myosin II phosphorylation and contractile force generation in vascular smooth muscle. However, relatively little is known about the contribution of this kinase to gastrointestinal smooth muscle contraction. The addition of a recombinant version of ZIPK that lacked the leucine zipper domain to permeabilized ileal strips evoked a Ca2+-independent contraction and resulted in myosin regulatory light chain diphosphorylation at Ser19 and Thr18. Neither Ca2+-independent force development nor myosin regulatory light chain phosphorylation was elicited by the addition of kinase-dead ZIPK to the ileal strips. The sensitivity of ZIPK-induced contraction to various kinase inhibitors was similar to the in vitro sensitivity of purified ZIPK to these inhibitors. Staurosporine was the most effective ZIPK inhibitor, with a Ki value calculated to be 2.6 +/- 0.3 micromol/L. Through the use of specific kinase inhibitors, we determined that Rho-associated protein kinase and Ca2+/phospholipid-dependent protein kinase (protein kinase C) do not mitigate ZIPK-induced contraction in ileum. Our findings support a role for ZIPK in Ca2+ independent contractile force generation in gastrointestinal smooth muscle. PMID- 17464352 TI - BMPs regulate differentiation of a putative visceral endoderm layer within human embryonic stem-cell-derived embryoid bodies. AB - Human embryonic stem cells (HESCs), pluripotent cells derived from the inner cell mass (ICM) of human blastocysts, represent a novel tool for the study of early human developmental events. When cultured in suspension with serum, HESCs form spherical structures resembling embryoid bodies (EBs). We show that differentiation of HESCs within EBs occurs radially, with central cells then undergoing apoptosis in association with EB cavitation. Cells within the outer layer of cavitating EBs display stage-specific immunoreactivity to pan-keratin, cytokeratin-8, GATA6, alpha-fetoprotein, and transthyretin specific antibodies, and hybridization to disabled-2, GATA4, and GATA6 specific riboprobes. Transmission electron microscopy of these cells reveals clathrin-coated micropinocytotic vesicles, microvilli, and many vacuoles, a phenotype consistent with mouse visceral endoderm (VE) rather than mouse definitive or parietal endoderm. When cultured in media supplemented with the BMP inhibitor noggin, or in the absence of serum, HESC derivatives do not develop the mouse VE-like phenotype. The addition of BMP-4 to noggin-treated HESCs cultured in serum or in serum-free conditions reconstituted development of the VE-like phenotype. These data demonstrate that human EBs undergo developmental events similar to those of mouse EBs and that in vitro BMP signalling induces derivatives of the human ICM to express a phenotype similar to mouse VE. PMID- 17464353 TI - An antibody-conjugated internalizing quantum dot suitable for long-term live imaging of cells. AB - Quantum dots (QD) are fluorescent semiconductor nanocrystals that are emerging as superior alternatives to the conventional organic dyes used in biological applications. Although QDs offer several advantages over conventional fluorescent dyes, including greater photostability and a wider range of excitation and (or) emission wavelengths, their toxicity has been an issue in its wider use as an analytic, diagnostic and therapeutic tool. We prepared a conjugate QD with an internalizing antibody and demonstrated that the QD-antibody conjugate is efficiently internalized into cells and is visible even after multiple divisions. We demonstrate that the internalized QD is nontoxic to cells and provides a sensitive tool for long-term molecular imaging. PMID- 17464354 TI - Substitution of methionine 435 with leucine, isoleucine, and serine in tumor necrosis factor alpha converting enzyme inactivates ectodomain shedding activity. AB - Tumor necrosis factor alpha (TNF-alpha) converting enzyme (TACE) is a zinc metalloprotease that has emerged as a general sheddase, which is responsible for ectodomain release of numerous membrane proteins, including the proinflammatory cytokine TNF-alpha, the leukocyte adhesin L-selectin and epidermal growth factor receptor ligand-transforming growth factor alpha (TGF-alpha), and related family members. Structurally, TACE belongs to a large clan of proteases, designated the metzincins, because TACE possesses a conserved methionine (Met435), frequently referred to as the met-turn residue, in its active site. A vital role of this residue in the function of TACE is supported by the fact that cells expressing the M435I TACE variant are defective in ectodomain shedding. However, the importance of Met435 in TACE appears to be uncertain, since another metzincin, matrix metalloprotease-2, has been found to be enzymatically fully active with either leucine or serine in place of its met-turn residue. We constructed TACE mutants with leucine or serine in place of Met435 to further examine the role of the met-turn residue in TACE-mediated ectodomain cleavage. Similar to the M435I TACE mutant, both the M435L and M435S constructs are defective in cleaving transmembrane TNF-alpha, TGF-alpha, and L-selectin. Comparative modeling and dynamic computation detected structural perturbations, which resulted in higher energy, in the catalytic zinc complexes of the Met435 TACE mutants compared with that in the wild-type enzyme. Thus, Met435 serves to maintain the stability of the catalytic center of TACE for the hydrolysis of peptide bonds in substrates. PMID- 17464355 TI - Ovotransferrin expression and release by chicken cell lines infected with Marek's disease virus. AB - Mammals posses both serum transferrin and lactoferrin, whose functions are taken over in birds by ovotransferrin, displaying both iron transport and antibacterial activities. Ovotransferrin also exerts antiviral activity towards Marek's disease virus, an avian member of the herpes family of viruses. This virus infects lymphoid organs and induces the transcription of ovotransferrin in infected chicken embryo fibroblasts. However, it has not yet been established whether ovotransferrin gene transcription is linked to the release of the protein outside the cells or whether ovotransferrin expression and release also occurs in chicken lymphoblastoid cells in which the Marek's disease viral genome is integrated. Our results indicate that both serum and egg-white isoforms of ovotransferrin are expressed and released in the supernatants of chicken embryo fibroblast and lymphoblastoid cells in the absence of infection. Viral infection of chicken embryo fibroblasts caused a slight increase of ovotransferrin release, whereas viral reinfection of lymphoblastoid cells caused a remarkable ovotransferrin release in a virus concentration-dependent manner. These findings suggest that ovotransferrin release in vivo may play a crucial role in protecting the whole organism from viral infection spreading, and support the hypothesis that the antiviral activity of ovotransferrin is an important part of the innate immune response in birds, resembling the antiviral activity of lactoferrin in mammals. PMID- 17464356 TI - An improved high throughput protein-protein interaction assay for nuclear hormone receptors. AB - The Glutathione-S-Transferase (GST) "pulldown" assay has been used extensively to assay protein interactions in vitro. This methodology has been especially useful for investigating the interactions of nuclear hormone receptors with a wide variety of their interacting partners and coregulatory proteins. Unfortunately, the original GST-pulldown technique relies on multiple binding, washing and elution steps performed in individual microfuge tubes, and requires repeated centrifugation, aspiration, and suspension steps. This type of batch processing creates a significant liquid handling bottleneck, limiting the number of sample points that can be incorporated into one experiment and producing inherently less efficient washing and elution than would a flow-through methodology. In this manuscript, we describe the adaptation of this GST-pulldown assay to a 96-well filter plate format. The use of a multi-well filter plate makes it possible to assay more samples in significantly less time using less reagents and more efficient sample processing than does the traditional single tube assay. PMID- 17464357 TI - Natural disordered sequences in the amino terminal domain of nuclear receptors: lessons from the androgen and glucocorticoid receptors. AB - Steroid hormones are a diverse class of structurally related molecules, derived from cholesterol, that include androgens, estrogens, progesterone and corticosteroids. They represent an important group of physiologically active signalling molecules that bind intracellular receptor proteins and regulate genes involved in developmental, reproductive and metabolic processes. The receptor proteins share structurally and functionally related ligand binding and DNA binding domains, but possess distinct N-terminal domains (NTD) of unique length and amino acids sequence. The NTD contains sequences important for gene regulation, exhibit structure plasticity and are likely to contribute to the specificity of the steroid hormone/receptor response. PMID- 17464358 TI - Visualizing the action of steroid hormone receptors in living cells. AB - Transcription controlled by Steroid Hormone Receptors (SHRs) plays a key role in many important physiological processes like organ development, metabolite homeostasis, and response to external stimuli. Understandably, the members of this family have drawn a lot of attention from the scientific community since their discovery, four decades ago. Still, after many years of research we are only beginning to unravel the complex nature of these receptors. The pace at which we do has improved significantly in recent years with the discovery of genetically encoded fluorescent probes, and the accompanying revival of biophysical approaches that allow more detailed study of SHRs. Here, we will look into the different aspects of SHR signalling, and discuss how biophysical techniques have contributed to visualizing their function in their native context, the living cell. PMID- 17464359 TI - gemini pollen 2, a male and female gametophytic cytokinesis defective mutation. AB - Gametophytic cytokinesis is essential for the development and function of the male and female gametophytes. We have previously described the isolation and characterisation of the gemini pollen 1 (gem1) that acts gametophytically to disturb asymmetric division and cytokinesis at pollen mitosis I in Arabidopsis. Here we describe the genetic and cytological analysis of an independent gametophytic mutant, gem2, with similar characteristics to gem1, but which maps to a different genetic locus. gem2 shows reduced genetic transmission through both male and female gametes and leads to the production of divided or twin celled pollen. Developmental analysis revealed that gem2 does not affect karyokinesis at pollen mitosis I, but leads to repositioning of the cell plate and partial or complete failure of cytokinesis, resulting in symmetrical divisions or binucleate pollen grains respectively. Symmetrical divisions lead to altered pollen cell fate with both sister cells displaying vegetative cell fate. Moreover, we demonstrate that the predominant female defect in gem2 is a lack of cellularization of the embryo sac during megagametogenesis. GEM2 therefore defines an independent genetic locus that is involved in the correct specification of both male and female gametophytic cytokinesis. PMID- 17464360 TI - Supercritical carbon dioxide: putting the fizz into biomaterials. AB - This paper describes recent progress made in the use of high pressure or supercritical fluids to process polymers into three-dimensional tissue engineering scaffolds. Three current examples are highlighted: foaming of acrylates for use in cartilage tissue engineering; plasticization and encapsulation of bioactive species into biodegradable polyesters for bone tissue engineering; and a novel laser sintering process used to fabricate three dimensional biodegradable polyester structures from particles prepared via a supercritical route. PMID- 17464361 TI - Three-Dimensional Bioactive and Biodegradable Scaffolds Fabricated by Surface Selective Laser Sintering. PMID- 17464362 TI - DIFFERENTIAL ATTACHMENT RESPONSES OF MALE AND FEMALE INFANTS TO FRIGHTENING MATERNAL BEHAVIOR: TEND OR BEFRIEND VERSUS FIGHT OR FLIGHT? AB - Taylor and colleagues (2000) proposed that males tend to display fight or flight responses to threat while females are more likely to display affiliative "tend or befriend" responses. In light of this hypothesis, gender differences in infant attachment behaviors were examined in a sample of 65 low-income mother-infant dyads, half of whom were referred to a home-based intervention service because of concerns about the quality of caregiving. Attachment behaviors were assessed in the Ainsworth Strange Situation when infants were 18 months old, and maternal behaviors were coded both for frightened or frightening behaviors, using the Main and Hesse (1992) coding inventory, and for disrupted affective communication using the Atypical Maternal Behavior Instrument for Assessment and Classification assessment tool (AMBIANCE; Lyons-Ruth, Bronfman, & Parsons, 1999). Results indicated that as maternal behavior became more frightening, female infants tended to approach their mothers more than male infants. These gender differences in response to maternal frightening behavior also were evident in the clinically referred subsample. The results suggest that gender-based differences in tendencies to show affiliative behaviors to threat may complicate interpretation of attachment behavior in clinical contexts. PMID- 17464363 TI - DISORGANIZED INFANT ATTACHMENT STRATEGIES AND HELPLESS-FEARFUL PROFILES OF PARENTING: INTEGRATING ATTACHMENT RESEARCH WITH CLINICAL INTERVENTION. AB - In this article, recent research on parenting behaviors associated with infant attachment disorganization is summarized and applied to a parent-infant psychotherapy case. Both hostile/self-referential and helpless-fearful patterns of parentingare described and viewed theoretically as alternate aspects of a single hostile-helpless internal working model of attachment relationships. The case material focuses on the more subtle and harder to identify manifestations of a helpless-fearful parental stance. Some attachment-related treatment guidelines for working with a hostile-helpless parenting stance are suggested, including challenging the hostile-helpless model implicitly in the qualities of the therapist's approach to the parent, explicitly articulating the hostile-helpless bind with the parent, increasing the parent's openness to a wider range of affective experience, differentiating attachment-related needs from other communications of the baby, and developing new skills for balancing the needs of the self and the needs of the other in interaction with the baby. PMID- 17464365 TI - Synthesis of Hydrodipyrrins Tailored for Reactivity at the 1- and 9-Positions. AB - A collection of 33 hydrodipyrrins (9 targets, 21 intermediates, and 3 byproducts) has been prepared. The hydrodipyrrins (dihydrodipyrrins, tetrahydrodipyrrins, and hexahydrodipyrrins) contain a pyrrole ring and a geminal-dimethyl substituted 1 pyrroline (or pyrrolidine) ring. The alpha-substituents on the pyrrole ring (H, Br, CHO) and pyrroline ring (H, CH(3), CH(OR)(2), OMe, SMe) provide different reactivity combinations (Nu(-), E(+)) and 0, 1, or 2 carbon atoms (which can give rise to the bridging meso-carbons in hydroporphyrins). Straightforward access to various hydrodipyrrins should facilitate development of syntheses of diverse hydroporphyrins. PMID- 17464364 TI - Dynamic changes in Histone H3 lysine 9 acetylation localization patterns during neuronal maturation require MeCP2. AB - Mutations within the gene encoding methyl CpG binding protein 2 (MECP2) cause the autism-spectrum neurodevelopmental disorder Rett Syndrome (RTT). MECP2 recruits histone deacetylase to methylated DNA and acts as a long-range regulator of methylated genes. Despite ubiquitous MECP2 expression, the phenotype of RTT and the Mecp2-deficient mouse is largely restricted to the postnatal brain. Since Mecp2-deficient mice have a defect in neuronal maturation, we sought to understand how MECP2/Mecp2 mutations globally affect histone modifications during postnatal brain development by an immunofluorescence approach. Using an antibody specific to acetylated histone H3 lysine 9 (H3K9ac), a bright punctate nuclear staining pattern was observed as MECP2 expression increased in early postnatal neuronal nuclei. As neurons matured in juvenile and adult brain samples, the intensity of H3K9ac staining was reduced. Mecp2-deficient mouse and RTT cerebral neurons lacked this developmental reduction in H3K9ac staining compared to age matched controls, resulting in a significant increase in neuronal nuclei with bright H3K9ac punctate staining. In contrast, trimethylated histone H3 lysine 9 (H3K9me3) localized to heterochromatin independent of MeCP2, but showed significantly reduced levels in Mecp2 deficient mouse and RTT brain. Autism brain with reduced MECP2 expression displayed similar histone H3 alterations as RTT brain. These observations suggest that MeCP2 regulates global histone modifications during a critical postnatal stage of neuronal maturation. These results have implications for understanding the molecular pathogenesis of RTT and autism in which MECP2 mutation or deficiency corresponds with arrested neurodevelopment. PMID- 17464366 TI - INTRODUCTION: Immune mechanisms of neurodegeneration. PMID- 17464367 TI - Interactions between retroactive-interference and context-mediated treatments that impair pavlovian conditioned responding. AB - In Pavlovian fear conditioning, context-mediated decrements in conditioned responding (e.g., the US preexposure effect) can counteract competition between cues trained together (e.g., overshadowing). Two experiments were conducted using rats in a conditioned lick suppression preparation to determine whether context mediated competition also counteracts competition between c ues trained separately (retroactive interference, or RI). In Experiment 1, a combination of degraded contingency and RI treatments produced less of a decrement in conditioned responding than did either of those treatments alone. Experiment 2 showed that RI treatment attenuates the normally deleterious effect of trial massing. The results suggest that empirical similarities are shared by interference between cues trained apart and competition between cues trained together. PMID- 17464368 TI - Older Workers: Who are the working poor in the U.S.? AB - With data from the 2000 Health and Retirement Study (HRS), the purpose of this study was to provide a profile of older workers who live poverty, and to compare the demographic, financial, employment, and health attributes of such individuals to similar persons not living in poverty. This study found that 3.5% of employed individuals between the ages of 51 and 61 belonged to the class of working poor. The results of the multivariate logistic regression analyses indicated that the older working poor were more likely to be non-White, less educated, non-married, and had lower levels of net worth than the working non-poor. They were more likely to be employed part time and were less likely to be covered by employee sponsored health insurance. PMID- 17464369 TI - Quality assurance in 3D dosimetry by optical-CT. PMID- 17464370 TI - A dual-purpose CCD based micro-optical-CT scanning system. PMID- 17464371 TI - Microwave-Enhanced High-Speed Fluorous Synthesis. AB - Increasing reaction speed and simplifying product purification are two major ways to improve the efficiency of organic synthesis. A new technology for high-speed solution-phase synthesis has been developed by combination of microwave heating and fluorous purification. This review describes different techniques for microwave-enhanced fluorous synthesis and their applications in Pd-catalyzed cross-coupling reactions, free-radical reactions, multicomponent reactions, and compound library synthesis. PMID- 17464372 TI - Palladium-Catalyzed Buchwald-Hartwig Type Amination of Fluorous Arylsulfonates. AB - Palladium-catalyzed cross-coupling reactions of aryl perfluorooctanesulfonates with amines are introduced. Application of the fluorous tag in multistep synthesis of triaryl-substituted pyrimidine is also described. PMID- 17464378 TI - Resource use study in COPD (RUSIC): a prospective study to quantify the effects of COPD exacerbations on health care resource use among COPD patients. AB - BACKGROUND: There is increasing interest in health care resource use (HRU) in Canada, particularly in resources associated with acute exacerbations of chronic obstructive pulmonary disease (COPD). OBJECTIVE: To identify HRU due to exacerbations of COPD. METHODS: A 52-week, multicentre, prospective, observational study of HRU due to exacerbations in patients with moderate to severe COPD was performed. Patients were recruited from primary care physicians and respirologists in urban and rural centres in Canada. RESULTS: In total, 524 subjects (59% men) completed the study. Their mean age was 68.2+/-9.4 years, with a forced expiratory volume in 1 s of 1.01+/-0.4 L. Patients had significant comorbidities. There were 691 acute exacerbations of COPD, which occurred in 53% of patients: 119 patients (23%) experienced one acute exacerbation, 70 patients (13%) had two acute exacerbations and 89 patients (17%) had three or more acute exacerbations. Seventy-five patients were admitted to hospital, with an average length of stay of 13.2 days. Fourteen of the patients spent time in an intensive care unit (average length of stay 5.6 days). Factors associated with acute exacerbations of COPD included lower forced expiratory volume in 1 s (P<0.001), high number of respiratory medications prescribed (P=0.037), regular use of oral corticosteroids (OCSs) (P=0.008) and presence of depression (P<0.001). Of the 75 patients hospitalized, only 53 received OCSs, four received referral for rehabilitation and 15 were referred for home care. CONCLUSIONS: The present study showed a high prevalence of COPD exacerbations, which likely impacted on HRU. There was evidence of a lack of appropriate management of exacerbations, especially with respect to use of OCSs, and referral for pulmonary rehabilitation and home care. PMID- 17464379 TI - Effects of changes in lung volume on oscillatory flow rate during high-frequency chest wall oscillation. AB - BACKGROUND: The effectiveness of high-frequency chest wall oscillation (HFCWO) in mucolysis and mucous clearance is thought to be dependant on oscillatory flow rate (Fosc). Therefore, increasing Fosc during HFCWO may have a clinical benefit. OBJECTIVES: To examine effects of continuous positive airway pressure (CPAP) on Fosc at two oscillation frequencies in healthy subjects and patients with airway obstruction. METHODS: Five healthy subjects and six patients with airway obstruction underwent 12 randomized trials of HFCWO (CPAP levels of 0 cm H2O, 2 cm H2O, 4 cm H2O, 6 cm H2O, 8 cm H2O and 10 cm H2O at frequencies of 10 Hz and 15 Hz) within a body plethysmograph, allowing measurements of changes in lung volume. Fosc was measured by reverse plethysmography using a 20 L isothermic chamber near the mouth. At the end of each randomized trial, an inspiratory capacity manoeuvre was used to determine end-expiratory lung volume (EELV). RESULTS: EELV increased significantly (P<0.05) with each level of CPAP regardless of oscillation frequency. Fosc also significantly increased with CPAP (P<0.05) and it was correlated with EELV (r=0.7935, P<0.05) in obstructed patients but not in healthy subjects (r=0.125, P=0.343). There were no significant differences in perceived comfort across the levels of CPAP. CONCLUSIONS: Significant increases in Fosc with CPAP-induced increases in lung volume were observed, suggesting that CPAP may be useful as a therapeutic adjunct in patients who have obstructive airway disease and who require HFCWO. PMID- 17464377 TI - Recommendations on chronic constipation (including constipation associated with irritable bowel syndrome) treatment. AB - While chronic constipation (CC) has a high prevalence in primary care, there are no existing treatment recommendations to guide health care professionals. To address this, a consensus group of 10 gastroenterologists was formed to develop treatment recommendations. Although constipation may occur as a result of organic disease, the present paper addresses only the management of primary CC or constipation associated with irritable bowel syndrome. The final consensus group was assembled and the recommendations were created following the exact process outlined by the Canadian Association of Gastroenterology for the following areas: epidemiology, quality of life and threshold for treatment; definitions and diagnostic criteria; lifestyle changes; bulking agents and stool softeners; osmotic agents; prokinetics; stimulant laxatives; suppositories; enemas; other drugs; biofeedback and behavioural approaches; surgery; and probiotics. A treatment algorithm was developed by the group for CC and constipation associated with irritable bowel syndrome. Where possible, an evidence-based approach and expert opinions were used to develop the statements in areas with insufficient evidence. The nature of the underlying pathophysiology for constipation is often unclear, and it can be tricky for physicians to decide on an appropriate treatment strategy for the individual patient. The myriad of treatment options available to Canadian physicians can be confusing; thus, the main aim of the recommendations and treatment algorithm is to optimize the approach in clinical care based on available evidence. PMID- 17464380 TI - An empirical continuous positive airway pressure trial for suspected obstructive sleep apnea. AB - BACKGROUND: Standard practice in obstructive sleep apnea (OSA) management requires that a positive diagnostic, overnight polysomnography (PSG) test be obtained before initiating treatment. However, long waiting times due to lack of access to PSG testing facilities may delay the initiation of definitive treatment for OSA. OBJECTIVES: To evaluate the response of patients who had a high clinical suspicion for OSA and who were waiting for a PSG test to an empirical continuous positive airway pressure (CPAP) trial. METHODS: A retrospective study of all patients who had been offered empirical CPAP therapy for suspected OSA was conducted. After outpatient assessment, 183 patients with a high pretest probability of having OSA began empirical CPAP testing using an arbitrary CPAP pressure. The presence of OSA, the accuracy of empirical CPAP pressure prescription, the adherence to empirical CPAP and the improvement in daytime somnolence were evaluated at the time of PSG. RESULTS: Of 183 patients on a CPAP trial, 91% had OSA, which was at least moderate (more than 15 apneas and hypopneas per hour of sleep) in 75% of the patients. Eighty per cent of the patients had significant daytime somnolence (Epworth Sleepiness Scale [ESS] greater than 10, mean +/- SD ESS 14+/-5), which improved with CPAP (ESS 9.0+/-5, P<0.01). In 40% of the patients, the arbitrary CPAP pressure was lower than that determined by manual titration. Adherence to a trial of CPAP (longer than 2 h/night) predicted OSA with a sensitivity of 82% and a specificity of 41%; the positive and negative predictive values were 92% and 22%, respectively. CONCLUSIONS: At the time of PSG testing, OSA was present in 91% of the patients who had received empirical CPAP. An empirical CPAP provided satisfactory interim treatment for excessive somnolence, despite the fact that the CPAP pressure was suboptimal in 40% of the patients. PMID- 17464381 TI - Pleuroparenchymal lung disease secondary to nonoccupational exposure to vermiculite. AB - An unusual case of pleuroparenchymal lung disease caused by the inhalation of vermiculite dust, presumably containing asbestos fibers is described. The uniqueness of the case lies in the very indirect nature of exposure -- the wife of a factory owner, rather than a worker exposed to asbestos, whose factory manufactured vermiculite. The present case illustrates the importance of taking careful occupational histories of all household members when presented with a patient whose chest radiograph exhibits features consistent with asbestos exposure. PMID- 17464382 TI - Erlotinib-associated acute pneumonitis: report of two cases. AB - Two cases of erlotinib-associated acute pneumonitis are described. The first patient was started on erlotinib treatment for metastatic non-small cell lung cancer. The second patient was treated with erlotinib for metastatic adenocarcinoma of unknown origin. Both patients developed dyspnea and hypoxemia five to six days after initiation of erlotinib treatment. In both cases, computed tomography scan of the chest showed extensive bilateral ground-glass infiltrates consistent with pneumonitis. In both patients, acute pneumonitis resulted in respiratory failure requiring intubation and mechanical ventilation. Diffuse alveolar hemorrhage was excluded by bronchoscopy in both cases. Bronchoalveolar lavage cultures were negative. Erlotinib treatment was stopped and both patients were treated with corticosteroids. The first patient improved gradually and finally was discharged to a rehabilitation centre, but unfortunately the second patient died of Klebsiella sepsis. Naranjo causality scale in both cases suggested a probable association between erlotinib and pneumonitis. Literature on erlotinib-associated pneumonitis is sparse. The clinical presentation and radiographic findings of erlotinib-associated acute pneumonitis are described. PMID- 17464383 TI - Severe chronic cough after Lap-Band gastric surgery. AB - Respirologists are often consulted in cases of chronic cough. Most patients have diagnoses of asthma, upper airway cough or gastroesophageal reflux disease. The present case describes a newly recognized cause of a cough in a patient who had undergone laparoscopic adjustable gastric banding for weight loss. The cough was persistent and severe for several months, and did not respond to any therapeutic interventions. Imaging studies were not helpful in making the diagnosis, which was confirmed by deflating the Lap-Band, thereby eliminating the cough. As more patients undergo bariatric surgery, physicians must anticipate and be familiar with potential complications. PMID- 17464384 TI - Circular and linear dichroism of proteins. AB - Circular dichroism (CD) is an important technique in the structural characterisation of proteins, and especially for secondary structure determination. The CD of proteins can be calculated from first principles using the so-called matrix method, with an accuracy which is almost quantitative for helical proteins. Thus, for proteins of unknown structure, CD calculations and experimental data can be used in conjunction to aid structure analysis. Linear dichroism (LD) can be calculated using analogous methodology and has been used to establish the relative orientations of subunits in proteins and protein orientation in an environment such as a membrane. However, simple analysis of LD data is not possible, due to overlapping transitions. So coupling the calculations and experiment is an important strategy. In this paper, the use of LD for the determination of protein orientation and how these data can be interpreted with the aid of calculations, are discussed. We review methods for the calculation of CD spectra, focusing on semiempirical and ab initio parameter sets used in the matrix method. Lastly, a new web interface for online CD and LD calculation is presented. PMID- 17464385 TI - Luminescent chemosensors based on semiconductor quantum dots. AB - Semiconductor quantum dots are inorganic nanoparticles with unique photophysical properties. In particular, their huge one- and two-photon absorption cross sections, tunable emission bands and excellent photobleaching resistances are stimulating the development of luminescent probes for biomedical imaging and sensing applications. Indeed, electron and energy transfer processes can be designed to switch the luminescence of semiconductor quantum dots in response to molecular recognition events. On the basis of these operating principles, the presence of target analytes can be transduced into detectable luminescence signals. In fact, luminescent chemosensors based on semiconductor quantum dots are starting to be developed to detect small molecules, monitor DNA hybridization, assess protein-ligand complementarities, test enzymatic activity and probe pH distributions. Although fundamental research is still very much needed to understand further the fundamental factors regulating the behavior of these systems and refine their performance, it is becoming apparent that sensitive probes based on semiconductor quantum dots will become invaluable analytical tools for a diversity of applications in biomedical research. PMID- 17464386 TI - New theoretical investigations of the photodissociation of ozone in the Hartley, Huggins, Chappuis, and Wulf bands. AB - We review recent theoretical studies of the photodissociation of ozone in the wavelength region from 200 nm to 1100 nm comprising four major absorption bands: Hartley and Huggins (near ultraviolet), Chappuis (visible), and Wulf (near infrared). The quantum mechanical dynamics calculations use global potential energy surfaces obtained from new high-level electronic structure calculations. Altogether nine electronic states are taken into account in the theoretical descriptions: four 1A', two 1A'', one 3A' and two 3A'' states. Of particular interest is the analysis of diffuse vibrational structures, which are prominent in all absorption bands, and their dynamical origin and assignment. Another focus is the effect of non-adiabatic coupling on lifetimes in the excited states and on the population of the specific electronic product channels. PMID- 17464387 TI - Torsional anharmonicity in the conformational analysis of tryptamine. AB - In this paper we calculate the relative conformer populations of the tryptamine molecule. Our approach combines high level electronic structure conformer energies with harmonic frequencies and an anharmonic treatment of the torsional motions using the torsional path integral Monte Carlo method. We have developed a 3-D potential energy surface as a function of the torsional coordinates at the B3LYP/6-31+G(d) level using 2535 grid points. Eight conformers of tryptamine were found to be significantly populated at 430 K as opposed to the experimental observation of seven. This, along with further comparisons with various experimental data, leads us to suppose that conformer interconversion occurs during the cooling phases of many of the experiments. The ordering of the calculated populations fits well with available experimental data. Torsional anharmonicity is found to affect conformer populations more significantly at 430 K than at 100 K (although overall the effects are small), while quantum mechanical effects are not important at either temperature. PMID- 17464388 TI - Conical intersections involving the dissociative 1pisigma* state in 9H-adenine: a quantum chemical ab initio study. AB - The conical intersections of the dissociative 1pisigma* excited state with the lowest 1pipi* excited state and the electronic ground state of 9H-adenine have been investigated with multireference electronic structure calculations. Adiabatic and quasidiabatic potential energy surfaces and coupling elements were calculated as a function of the NH stretch coordinate of the azine group and the out-of-plane angle of the hydrogen atom, employing MultiReference Configuration Interaction (MRCI) as well as Complete-Active-Space Self-Consistent-Field (CASSCF) methods. Characteristic properties of the 1pipi*-1pisigma* and 1pisigma* S0 conical intersections, such as the diabatic-to-adiabatic mixing angle, the geometric phase of the adiabatic electronic wavefunctions, the derivative coupling, as well as adiabatic and diabatic transition dipole moment surfaces were investigated in detail. These data are a prerequisite for future quantum wavepacket simulations of the photodissociation and internal-conversion dynamics of adenine. PMID- 17464389 TI - Interaction of heteroboranes with biomolecules. Part 2. The effect of various metal vertices and exo-substitutions. AB - Icosahedral heteroboranes and especially metallacarboranes, which have recently been shown to act as potent HIV-1 protease inhibitors, are a unique class of chemical compounds with unusual properties, one of which is the formation of dihydrogen bonds with biomolecules. In this study, we investigate the effect of various metal vertices and exo-substitutions on several series of heteroboranes, including 11-vertex carborane cages [nido-7,8-C2B9Hn]n-13(n= 11,12,13), closo-1 SB11H11, closo-1-NB11H12, metal bis(dicarbollides)[3,3'-M (1,2 C2B9H11)2]n(M/n=Fe/2-, Co/1-, Ni/0) and fluoro (F), amino (NH2) and hydroxo (OH) derivatives of the metal bis(dicarbollides). Besides the properties of isolated systems (geometries, electronic properties and hydration), we study their interactions with a tetrapeptide, which models their biomolecular partner. Calculations have confirmed that the extra hydrogen in [nido-7,8-C2B9H12]- forms a bridge, which fluctuates between two stationary states. Using RESP-derived charges, it was ascertained that the negative charge of heteroboranes is located mainly on boron-bound hydrogens. An increase of the negative total charge (from 0 to -1 or -2) of heteroboranes yields an increase in the stabilisation energies of heteroborane[dot dot dot]peptide complexes and also a substantial increase in the hydration free energies of heteroboranes. Compared to the substitutions of metal vertices, the exo-substitutions of metallacarboranes cause a larger increase in stabilisation energies and a smaller increase in desolvation penalties. These two terms, stabilisation energies and desolvation penalties, contribute in opposite directions to the total heteroborane-biomolecule binding energy and must both be taken into account when designing new HIV-1 protease inhibitors. PMID- 17464390 TI - Computational investigation of the Bi lone-pairs in monoclinic bismuth triborate BiB3O6. AB - The Bi-O interactions and the Bi lone-pairs in monoclinic BiB3O6 are investigated with gradient-corrected hybrid B3PW density functional theory within the Gaussian orbital-based CO-LCAO scheme. The Bi 6s and O 2p orbitals contribute to both bonding and antibonding interactions below the Fermi level. The stereochemical activity of the Bi lone-pairs was found to have a major origination from the primary interaction for the Bi 6s-O 2p antibonding orbital. The Bi 6p orbitals are not critically responsible for the non-spherical shape of the Bi lone-pairs, although they indeed participate into the secondary interaction with the Bi 6s-O 2p antibonding states. It is also suggested that O 2p components within the Bi lone-pairs are dominantly significant for the optical responses of BiB3O6 over the Bi 6s components. PMID- 17464391 TI - High resolution rotational spectroscopy on D2O up to 2.7 THz in its ground and first excited vibrational bending states. AB - We present highly accurate laboratory measurements on the pure rotational spectrum of doubly deuterated water, D2O, in selected frequency regions from 10 GHz up to 2.7 THz. Around 140 rotational transitions in both the vibrational ground and first excited bending states (upsilon2=0,1) were measured in total, involving energy levels with unexcelled high J and Ka rotational quantum numbers. The data give valuable information for the spectroscopic analysis of this molecule. In the case of the light and non-rigid water molecule, standard methods for its analysis are limited due to large centrifugal distortion interactions. Here, we present a global analysis of rotational and rovibrational data of the upsilon2=0 and 1 states of D2O by means of an Euler expansion of the Hamiltonian. In addition to the newly measured pure rotational transitions, around 4000 rotational and rovibrational lines have been included from previous work. It was possible to reproduce the extensive dataset to nearly its experimental uncertainty. The improved predictive capability of the model compared to previous work will be demonstrated. PMID- 17464392 TI - Planar study of H2O+Cl<-->HO+HCl reactions. AB - The first four dimensional (4D) quantum scattering calculations on the tetra atomic H2O+Cl<-->HO+HCl reactions are reported. With respect to a full (6D) treatment, only the planar constraint and a fixed length for the HO spectator bond are imposed. This work explicitly accounts for the bending and local HO stretching vibrations in H2O, for the vibration of HCl and for the in-plane rotation of the H2O, HO and HCl molecules. The calculations are performed with the potential energy surface of Clary et al. and use a Born-Oppenheimer type separation between the motions of the light and the heavy nuclei. State-to-state cross sections are reported for a collision energy range 0-1.8 eV measured with respect to H2O+Cl. For the H2O+Cl reaction, present results agree with previous (3D) non planar calculations and confirm that excitation of the H2O stretching promotes more reactivity than excitation of the bending. New results are related to the rotation of the H2O molecule: the cross sections are maximal for planar rotational states corresponding to 1095%). The experimental results are additionally supported by AM1 semi-empirical calculations. PMID- 17464406 TI - Synthetic pores with sticky pi-clamps. AB - In this report, we describe design, synthesis, evaluation and molecular dynamics simulations of synthetic multifunctional pores with pi-acidic naphthalenediimide clamps. Experimental evidence is provided for the formation of unstable but inert, heterogeneous and acid-insensitive dynamic tetrameric pores that are sensitive to base and ionic strength. Blockage experiments reveal that the introduction of aromatic electron donor-acceptor interactions provides access to the selective recognition of pi-basic intercalators within the pore. This breakthrough is important for the application of synthetic pores as multianalyte sensors. PMID- 17464407 TI - 3,4,5-triarylisothiazoles via C-C coupling chemistry. AB - The regiocontrolled preparation of triarylisothiazoles is presented. 3-Halo-5 phenylisothiazole-4-carbonitriles, 1 (hal=Cl) and 18 (hal=I), are converted into the corresponding 4-bromo derivatives 5 (3-hal=Cl) and 24 (3-hal=I) via a Hunsdiecker strategy while the 4-iodo analogues 7 (3-hal=Cl) and 22 (3-hal=I) are prepared via a Hoffmann and Sandmeyer strategy. Regioselective Suzuki, Stille and Negishi reactions occur at C-4 with both the 4-bromo- and 4-iodoisothiazoles 5 and 7 , the latter being more reactive than the former. 3-Iodoisothiazoles 22 and 24 fail to give regiocontrolled Suzuki, Stille or Negishi couplings, however, 4 bromo-3-iodo-5-phenylisothiazole 24 gives the regiospecific palladium catalysed Ullmann-type reaction product 3,3'-bi(4-bromo-5-phenylisothiazole) 25 . Alkali hydrolysis of 3-chloro-4,5-diphenylisothiazole 8 gives the 3-hydroxy analogue 12 which is converted into 3-bromo-4,5-diphenylisothiazole 13 with POBr(3). 3 Bromoisothiazole 13 reacts with phenylzinc chloride to give 3,4,5 triphenylisothiazole 17 but fails to undergo effective Suzuki or Stille couplings. 3,5-Diphenylisothiazole-4-carbonitrile 26 is converted into the 4 bromo- and 4-iodo-3,5-diphenylisothiazoles 30 and 34 both of which are effective for Suzuki and Stille couplings. A series of triarylisothiazoles are prepared in this manner and fully characterised. PMID- 17464408 TI - Palladium-catalyzed sequential one-pot reaction of aryl bromides with O homoallylhydroxylamines: synthesis of N-aryl-beta-amino alcohols. AB - The palladium-catalyzed sequential one-pot N-arylation-carbo-amination-C arylation of O-homoallylhydroxylamines with two different aryl bromides provides rapid entry to differentially arylated N-aryl-3-arylmethylisoxazolidines in good yields with excellent diastereoselectivity. The obtained isoxazolidines can be reductively cleaved to cis-N-aryl-beta-amino alcohols in short times and in high yields at room temperature. PMID- 17464409 TI - Preparation of new axially chiral bridged 2,2'-bipyridines and pyridyl monooxazolines (pymox). Evaluation in copper(I)-catalyzed enantioselective cyclopropanation. AB - This work reports the synthesis of new axially chiral bridged 2,2'-bipyridines 1 and pyridylmonooxazolines (pymox) 2. The potential of these new axially chiral N,N-ligands was evaluated in asymmetric catalytic cyclopropanation of styrene derivatives 22a-c with diazoesters 21a,b. While 2,2'-bipyridines 1a-c afforded the corresponding cyclopropanes 23a-f in up to 65% ee, pymoxs 2a-e gave somewhat lower enantioselectivities (up to 53% ee). Both classes of ligands produced trans cyclopropanes 23a-f as the major isomer, although with modest diasteroselectivities (56 : 44 to 78 : 22). A structure-stereoselectivity relationship study of ligands 1 and 2 identified the chiral biaryl axis as being mostly responsible for the enantioselective performances of these ligands. PMID- 17464410 TI - Asymmetric three- and [2 + 1]-component conjugate addition reactions for the stereoselective synthesis of polysubstituted piperidinones. AB - The efficiency and stereoselectivity of the conjugate addition of lithium (Z)- or (E)-beta-amino ester enolates, generated by lithium amide conjugate addition to an alpha,beta-unsaturated ester or deprotonation of a beta-amino ester, respectively, to a range of alpha,beta-unsaturated acceptors has been investigated. Deprotonation of a beta-amino ester with LDA, followed by conjugate addition to a chiral alpha,beta-unsaturated oxazolidinone gives high 2,3-anti selectivity ( approximately 90% d.e.), with hydrogenolysis and purification to homogeneity generating stereodefined trisubstituted piperidinones as single stereoisomers. Asymmetric three-component couplings of alpha,beta-unsaturated esters and alkylidene malonates initiated by lithium amide conjugate addition proceeds with high levels of 2,3-anti stereoselectivity, with hydrogenolysis giving tetrasubstituted piperidinones. PMID- 17464411 TI - Mixing of peptides and electrophilic traps gives rise to potent, broad-spectrum proteasome inhibitors. AB - The synthesis and evaluation of hybrid proteasome inhibitors that contain structural elements of the known inhibitors bortezomib, epoxomicin and peptide vinyl sulfones is described. From the panel of 15 inhibitors some structure activity relationships can be deduced with regard to inhibitory activity in relation to peptide recognition element, inhibitor size and nature of the electrophilic trap. Further, the panel contains one of the most potent peptide based pan-proteasome inhibitors reported to date. PMID- 17464412 TI - Design and total synthesis of unnatural analogues of the sub-nanomolar SERCA inhibitor thapsigargin. AB - Thapsigargin is a densely oxygenated guaianolide which displays potent sarco/endoplasmic reticulum Ca(2+) ATPase (SERCA) binding affinities. The total syntheses of designed unnatural analogues of this important natural product are described. This article constitutes the chemical synthesis behind an ongoing project. Rational modifications have been made to the lactone region of thapsigargin in order to obtain derivatives for future structure-activity relationship studies. PMID- 17464413 TI - Carboxyketenes, methyleneketenes, vinylketenes, oxetanediones, ynols, and ylidic ketenes from Meldrum's acid derivatives. AB - It has been documented that 5-methylene-Meldrum's acid derivatives (1, 12 ) and their enols (2, 13) can undergo fragmentation to malonic anhydrides (4, 19 ), carboxyketenes (3, 16) and methyleneketene (5, 21 , 35 ), as well as cyclization to pyrrole-3-one and thiophene-3-one derivatives 11a,b (but not furan-3-ones 11c ) under the conditions of flash vacuum thermolysis (FVT). Here we report theoretical calculations at the B3LYP/6-311 + G(2d, p) and G3X(MP2) levels of theory, which allow a rationalization of these observations. The calculated activation barriers for these reactions are all of the order of 37-40 kcal mol( 1). Hydroxyacetylenes (alkynols) 7 are sometimes observed in FVT reactions of Meldrum's acid derivatives. Their formation is now explained as an FVT reaction of the carboxyketenes (e.g. 3-->7 and 32-->34) with a calculated activation barrier of ca. 39 kcal mol(-1). The cyclization of alkylamino- and alkylthio substituted methyleneketenes 8a,b to pyrrolone and thiophenone derivatives 11a,b is found to be energetically very feasible under FVT conditions, and even in some cases in solution, with activation barriers of 33-39 kcal mol(-1). This cyclization takes place via the fleeting ylidic ketene intermediates 9a,b,25, and 37a,b, which exist in very shallow energy minima. Alkoxy-substituted methyleneketenes 8c do not cyclize in this manner due to the rather high, but in principle not impossible, activation barriers for the initial 1,4-H shifts to the ylidic ketenes 9c (ca. 47 kcal mol(-1)). PMID- 17464414 TI - Reaction of perfluorocyclopentene with various carbon nucleophiles- heteroaromatic lithium reagents, enolate and phosphonium ylide. AB - The addition of heteroaromatic lithium reagents 2 to a THF solution of perfluorocyclopentene (1) provided preferentially the corresponding monosubstituted products 5, while the addition of 1 to 2 effectively gave the 1,2 disubstituted products 6 in good to excellent yields. The reaction of 1 with sodiomalonate 3 or phosphonium ylide 4 also proceeded smoothly to form the 1,3 disubstituted product 8 or 10 in high yield, respectively. PMID- 17464415 TI - An efficient route for the synthesis of a new class of pyrido[2,3-d]pyrimidine derivatives. AB - A new reaction of 4-arylidene-3-methylisoxazol-5(4H)-one or 4-arylidene-2 phenyloxazol-5(4H)-one with 2,6-diaminopyrimidin-4(3H)-one is described and a number of new pyrido[2,3-d]pyrimidine-4,7-dione derivatives are synthesized. This protocol has the advantages of good yields, broad substrate scope and simple work up. PMID- 17464416 TI - Synthesis and characterization of a new fluorescent probe for reactive oxygen species. AB - We report the development of a new fluorescence sensor for reactive oxygen species (ROS) based on a benzofurazan structure. The sensor, NBFhd, is initially non-fluorescent and reacts with peroxyl radicals by hydrogen transfer in an aqueous medium under neutral conditions to release the fluorescent N-methyl-4 amino-7-nitrobenzofurazan (NBF) and 1,4-benzoquinone. NBFhd shows excellent contrast and no interference in the region of cell autofluorescence and is a new tool to detect ROS in homogeneous and biological systems. PMID- 17464417 TI - Synthesis, characterization and cytolytic activity of alpha-helical amphiphilic peptide nanostructures containing crown ethers. AB - Many natural alpha-helical amphiphilic peptides are known to have lytic activity toward different cells. Herein, we describe the synthesis and the characterization of synthetic alpha-helical amphiphilic peptide nanostructures containing crown ethers, as well as the modulation of their cytolytic activity by adding different acidic dipeptide chains at the N- or C-terminus. PMID- 17464418 TI - A concise first total synthesis of narceine imide. AB - A concise and efficient total synthesis of alkaloid narceine imide is disclosed. The key steps are based upon the sequential construction of the isoindolinone template followed by metalation and coupling with an isoquinolinium salt. Subsequent E1cb elimination enables the creation of the arylmethylene unit with the concomitant formation of the dimethylaminoethyl chain and ultimate deprotection completes the synthesis of the natural product. PMID- 17464419 TI - Nonregular structure-property relationships for inclusion parameters of tert butylcalix[5]arene. AB - The effect of macrocycle size on the structure-property relationships was studied for inclusion compounds of tert-butylcalix[n]arenes (n=4,5) with volatile organic guests having various molecular size and group composition. Vapor-sorption isotherms, guest-inclusion stoichiometry and Gibbs energy, thermostability parameters and decomposition enthalpies were determined for host-guest compounds (clathrates) obtained using saturation of solid calixarene powder with guest vapor. The increase of the host macrocycle in the studied calixarene pair changes the observed structure-property relationship from the guest-binding selectivity mostly seen in inclusion Gibbs energy to the high sensitivity for guest structure in inclusion stoichiometry. The host with the larger macrocycle has more clathrates with stepwise formation and decomposition. Specific types of guest binding with solid hosts are discussed. PMID- 17464420 TI - New paradigms and ethics. PMID- 17464421 TI - Combating oncogene activation associated with retrovirus-mediated gene therapy of X-linked severe combined immunodeficiency. AB - A successful gene therapy clinical trial that also encountered serious adverse effects has sparked extensive study and debate about the future directions for retrovirus-mediated interventions. Treatment of X-linked severe combined immunodeficiency with an oncoretrovirus harboring a normal copy of the gammac gene was applied in two clinical trials, essentially curing 13 of 16 infants, restoring a normal immune system without the need for additional immune-related therapies. Approximately 3 years after their gene therapy, tragically, 3 of these children, all from the same trial, developed leukemia as a result of this experimental treatment. The current understanding of the mechanism behind this leukemogenesis involves three critical and cooperating factors, i.e., viral integration, oncogene activation, and the function of the therapeutic gene. In this review, we will explore the causes of this unwanted event and some of the possibilities for reducing the risk of its reoccurrence. PMID- 17464422 TI - Pipeline for macro- and microarray analyses. AB - The pipeline for macro- and microarray analyses (PMmA) is a set of scripts with a web interface developed to analyze DNA array data generated by array image quantification software. PMmA is designed for use with single- or double-color array data and to work as a pipeline in five classes (data format, normalization, data analysis, clustering, and array maps). It can also be used as a plugin in the BioArray Software Environment, an open-source database for array analysis, or used in a local version of the web service. All scripts in PMmA were developed in the PERL programming language and statistical analysis functions were implemented in the R statistical language. Consequently, our package is a platform independent software. Our algorithms can correctly select almost 90% of the differentially expressed genes, showing a superior performance compared to other methods of analysis. The pipeline software has been applied to 1536 expressed sequence tags macroarray public data of sugarcane exposed to cold for 3 to 48 h. PMmA identified thirty cold-responsive genes previously unidentified in this public dataset. Fourteen genes were up-regulated, two had a variable expression and the other fourteen were down-regulated in the treatments. These new findings certainly were a consequence of using a superior statistical analysis approach, since the original study did not take into account the dependence of data variability on the average signal intensity of each gene. The web interface, supplementary information, and the package source code are available, free, to non-commercial users at http://ipe.cbmeg.unicamp.br/pub/PMmA. PMID- 17464423 TI - TIMP-1 mediates the inhibitory effect of interleukin-6 on the proliferation of a hepatocarcinoma cell line in a STAT3-dependent manner. AB - The tissue inhibitor of metalloproteinases (TIMP)-1 is a multifunctional protein which is not only an inhibitor of matrix metalloproteinases (MMPs) but also to have a possible "cytokine-like" action. Here, we first compared mRNA expression of TIMP-1 and MMP-9 in BEL-7402 (a hepatocellular carcinoma cell line), L-02 (a normal liver cell line) and QSG-7701 (a cell line derived from peripheral tissue of liver carcinoma) using real-time quantitative RT-PCR. By evaluating the variation of the MMP-9/TIMP-1 ratio as an index of reciprocal changes of the expression of the two genes, we observed that the MMP-9/TIMP-1 ratio was about 13 and 5-fold higher in BEL-7402 than in L-02 and QSG-7701, respectively. Significantly, overexpression of TIMP-1 decreased the MMP-9/TIMP-1 ratio in BEL 7402 and then inhibited the cell growth to 60% and reduced the migration to about 30%. Meanwhile, our data showed that interleukin-6 (IL-6) (100 ng/mL) could also inhibited the cell growth of BEL-7402. Further studies indicated that TIMP-1 mediated the inhibitory effect of IL-6 on BEL-7402 cell proliferation in a STAT3 dependent manner, which could further accelerate the expression of the cyclin dependent kinase inhibitor p21. A dominant negative STAT3 mutant totally abolished IL-6-induced TIMP-1 expression and its biological functions. The present results demonstrate that TIMP-1 may be one of the mediators that regulate the inhibitory effect of IL-6 on BEL-7402 proliferation in which STAT3 signal transduction and p21 up-regulation also play important roles. PMID- 17464424 TI - Sleep deprivation reduces the lymphocyte count in a non-obese mouse model of type 1 diabetes mellitus. AB - The objective of the present study was to determine whether sleep deprivation (SD) would promote changes in lymphocyte numbers in a type 1 diabetes model (non obese diabetic, NOD, mouse strain) and to determine whether SD would affect female and male NOD compared to Swiss mice. The number of lymphocytes in peripheral blood after 24 and 96 h of SD (by multiple platform method) or equivalent period of time in home-cage controls was examined prior to the onset of diabetes. SD for 96 h significantly reduced lymphocytes in male Swiss mice compared to control (8.6 +/- 2.1 vs 4.1 +/- 0.7 10(3)/microL; P < 0.02). In male NOD animals, 24- and 96-h SD caused a significant decrease of lymphocytes compared to control (4.4 +/- 0.3 vs 1.6 +/- 0.5; P < 0.001 and 4.4 +/- 0.3 vs 0.9 +/- 0.1 10(3)/microL; P < 0.00001, respectively). Both 24- and 96-h SD induced a reduction in the number of lymphocytes in female Swiss (7.5 +/- 0.5 vs 4.5 +/- 0.5, 4.4 +/- 0.6 10(3)/microL; P < 0.001, respectively) and NOD mice (4 +/- 0.6 vs 1.8 +/- 0.2, 1.2 +/- 0.4 10(3)/microL; P < 0.01, respectively) compared to the respective controls. Loss of sleep induced lymphopenia in peripheral blood in both genders and strains used. Since many cases of autoimmunity present reduced numbers of lymphocytes and, in this study, it was more evident in the NOD strain, our results suggest that SD should be considered a risk factor in the onset of autoimmune disorders. PMID- 17464425 TI - Dimensionality of the premenstrual syndrome: confirmatory factor analysis of premenstrual dysphoric symptoms among college students. AB - Premenstrual syndrome and premenstrual dysphoric disorder (PMDD) seem to form a severity continuum with no clear-cut boundary. However, since the American Psychiatric Association proposed the research criteria for PMDD in 1994, there has been no agreement about the symptomatic constellation that constitutes this syndrome. The objective of the present study was to establish the core latent structure of PMDD symptoms in a non-clinical sample. Data concerning PMDD symptoms were obtained from 632 regularly menstruating college students (mean age 24.4 years, SD 5.9, range 17 to 49). For the first random half (N = 316), we performed principal component analysis (PCA) and for the remaining half (N = 316), we tested three theory-derived competing models of PMDD by confirmatory factor analysis. PCA allowed us to extract two correlated factors, i.e., dysphoric-somatic and behavioral-impairment factors. The two-dimensional latent model derived from PCA showed the best overall fit among three models tested by confirmatory factor analysis (chi(2)53 = 64.39, P = 0.13; goodness-of-fit indices = 0.96; adjusted goodness-of-fit indices = 0.95; root mean square residual = 0.05; root mean square error of approximation = 0.03; 90%CI = 0.00 to 0.05; Akaike's information criterion = -41.61). The items "out of control" and "physical symptoms" loaded conspicuously on the first factor and "interpersonal impairment" loaded higher on the second factor. The construct validity for PMDD was accounted for by two highly correlated dimensions. These results support the argument for focusing on the core psychopathological dimension of PMDD in future studies. PMID- 17464426 TI - Functional expression of kinin B1 and B2 receptors in mouse abdominal aorta. AB - Previous studies have shown that the vascular reactivity of the mouse aorta differs substantially from that of the rat aorta in response to several agonists such as angiotensin II, endothelin-1 and isoproterenol. However, no information is available about the agonists bradykinin (BK) and DesArg(9)BK (DBK). Our aim was to determine the potential expression of kinin B(1) and B(2) receptors in the abdominal mouse aorta isolated from C57BL/6 mice. Contraction and relaxation responses to BK and DBK were investigated using isometric recordings. The kinins were unable to induce relaxation but concentration-contraction response curves were obtained by applying increasing concentrations of the agonists BK and DBK. These effects were blocked by the antagonists Icatibant and R-715, respectively. The potency (pD(2)) calculated from the curves was 7.0 +/- 0.1 for BK and 7.3 +/- 0.2 for DBK. The efficacy was 51 +/- 2% for BK and 30 +/- 1% for DBK when compared to 1 microM norepinephrine. The concentration-dependent responses of BK and DBK were markedly inhibited by the arachidonic acid inhibitor indomethacin (1 microM), suggesting a mediation by the cyclooxygenase pathway. These contractile responses were not potentiated in the presence of the NOS inhibitor L-NAME (1 mM) or endothelium-denuded aorta, indicating that the NO pathway is not involved. We conclude that the mouse aorta constitutively contains B(1) and B(2) subtypes of kinin receptors and that stimulation with BK and DBK induces contractile effect mediated by endothelium-independent vasoconstrictor prostanoids. PMID- 17464427 TI - Activity of liver microsomal enzymes during the chronic phase of murine schistosomiasis. AB - The effects of schistosomiasis on microsomal enzymes were studied on post infection day 90 when accumulated damage and fibrosis are most intense but granulomatous reaction around the eggs harbored in the liver is smaller than during the earlier phases. Swiss Webster (SW) and DBA/2 mice of either sex (N = 12 per sex per group) were infected with 100 Schistosoma mansoni cercariae on postnatal day 10 and killed on post-infection day 90. Cytochrome P-450 (CYP) concentration and alkoxyresorufin-O-dealkylases (EROD, MROD, BROD, and PROD), p nitrophenol-hydroxylase (PNPH), coumarin-7-hydroxylase (COH), and UDP glucuronosyltransferase (UGT) activities were measured in hepatic microsomes. Age matched mice of the same sex and strain were used as controls. In S. mansoni infected mice, CYP1A- and 2B-mediated activities (control = 100%) were reduced in SW (EROD: male (M) 36%, female (F) 38%; MROD: M 38%, F 39%; BROD: M 46%, F 19%; PROD: M 50%, F 28%) and DBA/2 mice (EROD: M 64%, F 58%; MROD: M 60%; BROD: F 49%; PROD: M 73%) while PNPH (CYP2E1) was decreased in SW (M 31%, F 38%) but not in DBA/2 mice. COH did not differ between infected and control DBA/2 and UGT, a phase-2 enzyme, was not altered by infection. In conclusion, chronic S. mansoni infection reduced total CYP content and all CYP-mediated activities evaluated in SW mice, including those catalyzed by CYP2E1 (PNPH), CYP1A (EROD, MROD) and 2B (BROD, PROD). In DBA/2 mice, however, CYP2A5- and 2E1-mediated activities remained unchanged while total CYP content and activities mediated by other CYP isoforms were depressed during chronic schistosomiasis. PMID- 17464428 TI - In vitro evaluation of the cytotoxic and trypanocidal activities of Ampelozizyphus amazonicus (Rhamnaceae). AB - Ampelozizyphus amazonicus Ducke is a tree commonly found in the Amazon region and an extract of its stem bark is popularly used as an antimalarial and anti inflammatory agent and as an antidote to snake venom. Ursolic acid; five lupane type triterpenes: betulin, betulinic acid, lupenone, 3beta-hydroxylup-20(29)-ene 27,28-dioic acid, and 2alpha,3beta-dihydroxylup-20(29)-ene-27,28-dioic acid, and three phytosteroids: stigmasterol, sitosterol and campesterol, have been isolated from stem extracts of A. amazonicus Ducke. Their structures were characterized by spectral data including COSY and HMQC. In an in vitro biological screening of the isolated compounds, 3beta-hydroxylup-20(29)-ene-27,28-dioic acid was cytotoxic against the SKBR-3 human adenocarcinoma cell line (1 to 10 mg/mL), while 2alpha,3beta-dihydroxylup-20(29)-ene-27,28-dioic acid exhibited cytotoxicity against both SKBR-3 human adenocarcinoma and C-8161 human melanoma tumor cell lines (>0.1 mg/mL). In the present study, different extracts and some fractions of this plant were also investigated for trypanocidal activity due to the presence of pentacyclic triterpenes. The triterpene classes are potent against Trypanosoma cruzi. The bioassays were carried out using blood collected from Swiss albino mice by cardiac puncture during the parasitemic peak (7th day) after infection with the Y strain of T. cruzi. The results obtained showed that A. amazonicus is a potential source of bioactive compounds since its extracts and fractions isolated from it exhibited in vitro parasite lysis against trypomastigote forms of T. cruzi at concentrations >100 microg/mL. Fractions containing mainly betulin, lupenone, 3beta-hydroxylup-20(29)-ene-27,28-dioic acid, and 2alpha,3beta-dihydroxylup-20(29)-ene-27,28-dioic acid showed more activity than crude extracts. PMID- 17464429 TI - Effect of metabolic control on interferon-gamma and interleukin-10 production by peripheral blood mononuclear cells from type 1 and type 2 diabetic patients. AB - The objective of the present study was to evaluate the production of cytokines, interferon-gamma (INF-gamma) and interleukin-10 (IL-10), in cultures of peripheral blood mononuclear cells (PBMC) from type 1 and type 2 diabetic patients and to correlate it with inadequate and adequate metabolic control. We studied 11 type 1 and 13 type 2 diabetic patients and 21 healthy individuals divided into two groups (N = 11 and 10) paired by sex and age with type 1 and type 2 diabetic patients. The PBMC cultures were stimulated with concanavalin-A to measure INF-gamma and IL-10 supernatant concentration by ELISA. For patients with inadequate metabolic control, the cultures were performed on the first day of hospitalization and again after intensive treatment to achieve adequate control. INF-gamma levels in the supernatants of type 1 diabetic patient cultures were higher compared to type 2 diabetic patients with adequate metabolic control (P < 0.001). Additionally, INF-gamma and IL-10 tended to increase the liberation of PBMC from type 1 and 2 diabetic patients with adequate metabolic control (P = 0.009 and 0.09, respectively). The increased levels of INF-gamma and IL-10 released from PBMC of type 1 and 2 diabetic patients with adequate metabolic control suggest that diabetic control improves the capacity of activation and maintenance of the immune response, reducing the susceptibility to infections. PMID- 17464430 TI - Biphasic modulation of insulin receptor substrate-1 during goitrogenesis. AB - Insulin receptor substrate-1 (IRS-1) is the main intracellular substrate for both insulin and insulin-like growth factor I (IGF-I) receptors and is critical for cell mitogenesis. Thyrotropin is able to induce thyroid cell proliferation through the cyclic AMP intracellular cascade; however, the presence of either insulin or IGF-I is required for the mitogenic effect of thyroid-stimulating hormone (TSH) to occur. The aim of the present study was to determine whether thyroid IRS-1 content is modulated by TSH in vivo. Strikingly, hypothyroid goitrous rats, which have chronically high serum TSH levels (control, C = 2.31 +/ 0.28; methimazole (MMI) 21d = 51.02 +/- 6.02 ng/mL, N = 12 rats), when treated with 0.03% MMI in drinking water for 21 days, showed significantly reduced thyroid IRS-1 mRNA content. Since goiter was already established in these animals by MMI for 21 days, we also evaluated IRS-1 expression during goitrogenesis. Animals treated with MMI for different periods of time showed a progressive increase in thyroid weight (C = 22.18 +/- 1.21; MMI 5d = 32.83 +/- 1.48; MMI 7d = 31.1 +/- 3.25; MMI 10d = 33.8 +/- 1.25; MMI 14d = 45.5 +/- 2.56; MMI 18d = 53.0 +/- 3.01; MMI 21d = 61.9 +/- 3.92 mg, N = 9-15 animals per group) and serum TSH levels (C = 1.57 +/- 0.2; MMI 5d = 9.95 +/- 0.74; MMI 7d = 10.38 +/- 0.84; MMI 10d = 17.72 +/- 1.47; MMI 14d = 25.65 +/- 1.23; MMI 18d = 35.38 +/- 3.69; MMI 21d = 31.3 +/- 2.7 ng/mL, N = 9-15 animals per group). Thyroid IRS-1 mRNA expression increased progressively during goitrogenesis, being significantly higher by the 14th day of MMI treatment, and then started to decline, reaching the lowest values by the 21st day, when a significant reduction was detected. In the liver of these animals, however, a significant decrease of IRS-1 mRNA was detected after 14 days of MMI treatment, a mechanism probably involved in the insulin resistance that occurs in hypothyroidism. The increase in IRS-1 expression during goitrogenesis may represent an important event associated with the increased rate of cell mitosis promoted by TSH and indicates that insulin and IGF-I are important co-mitogenic factors in vivo, possibly acting through the activation of IRS-1. PMID- 17464431 TI - Cl- and regulation of pH by MDCK-C11 cells. AB - The interaction between H(+) extrusion via H(+)-ATPase and Cl(-) conductance was studied in the C11 clone of MDCK cells, akin to the intercalated cells of the collecting duct. Cell pH (pHi) was measured by fluorescence microscopy using the fluorescein-derived probe BCECF-AM. Control recovery rate measured after a 20 mM NH(4)Cl acid pulse was 0.136 +/- 0.008 pH units/min (dpHi/dt) in Na(+) Ringer and 0.032 +/- 0.003 in the absence of Na(+) (0 Na(+)). With 0 Na(+) plus the Cl(-) channel inhibitor NPPB (10 microM), recovery was reduced to 0.014 +/- 0.001 dpHi/dt. 8-Br-cAMP, known to activate CFTR Cl(-) channels, increased dpHi/dt in 0 Na(+) to 0.061 +/- 0.009 and also in the presence of 46 nM concanamycin and 50 microM Schering 28080. Since it is thought that the Cl(-) dependence of H(+) ATPase might be due to its electrogenic nature and the establishment of a +PD (potential difference) across the cell membrane, the effect of 10 microM valinomycin at high (100 mM) K(+) was tested in our cells. In Na(+) Ringer, dpHi/dt was increased, but no effect was detected in 0 Na+ Ringer in the presence of NPPB, indicating that in intact C11 cells the effect of blocking Cl(-) channels on dpHi/dt was not due to an adverse electrical gradient. The effect of 100 microM ATP was studied in 0 Na(+) Ringer solution; this treatment caused a significant inhibition of dpHi/dt, reversed by 50 microM Bapta. We have shown that H(+)-ATPase present in MDCK C11 cells depends on Cl(-) ions and their channels, being regulated by cAMP and ATP, but not by the electrical gradient established by electrogenic H(+) transport. PMID- 17464432 TI - A tribute to Samuel MacDonald McCann (1925-2007). In honor of one of the last pioneers of Neuroendocrinology. PMID- 17464433 TI - Mineralocorticoid modulation of central angiotensin-induced neuronal activity, water intake and sodium appetite. AB - Central angiotensin II (AngII) stimulates water and salt solution intake. Pretreatment with low-dose mineralocorticoid (DOCA) enhances this AngII-induced intake of salt solutions (the synergy theory) in Wistar and Sprague Dawley rats but not in Fischer rats. This response is mediated via the AT-1 receptor. Electrophysiological experiments using iontophoretic application of AngII and the AT-1 receptor-specific non-peptide antagonist losartan showed excitation of neurons in the preoptic/medial septum region of urethane-anesthetized male Wistar rats. DOCA pretreatment further enhances this neuronal excitation in response to AngII and reduces the responses to losartan. This generated the hypothesis that DOCA-enhanced AngII-induced neuronal excitation is the neural support for the synergy theory. AT-2 receptors modulate these intake responses depending on sodium in the diet, and diuretic-induced dehydration during pregnancy produces a higher salt intake in the offspring. AngII-induced salt and water intakes were tested in offspring from Sprague Dawley mothers with only 1.8% NaCl to drink in which half were treated with furosemide. The important observations were a) the AT-1 antagonist alone suppressed intakes in offspring from mothers not treated with furosemide, b) both AT-1 and AT-2 antagonists suppressed intakes in offspring from furosemide-treated mothers, and c) combined administration of AT-1 and AT-2 antagonists greatly suppressed water intake in offspring from mothers not treated with furosemide. These results suggest that AT-1 and AT-2 receptors have variable properties (receptor number and/or second messengers). Furthermore, the activity and function of these central AngII receptors depend on the background mineralocorticoid levels. The exact mechanism of this influence, however, remains to be determined. PMID- 17464434 TI - Water deprivation and the double- depletion hypothesis: common neural mechanisms underlie thirst and salt appetite. AB - Water deprivation-induced thirst is explained by the double-depletion hypothesis, which predicts that dehydration of the two major body fluid compartments, the extracellular and intracellular compartments, activates signals that combine centrally to induce water intake. However, sodium appetite is also elicited by water deprivation. In this brief review, we stress the importance of the water depletion and partial extracellular fluid-repletion protocol which permits the distinction between sodium appetite and thirst. Consistent enhancement or a de novo production of sodium intake induced by deactivation of inhibitory nuclei (e.g., lateral parabrachial nucleus) or hormones (oxytocin, atrial natriuretic peptide), in water-deprived, extracellular-dehydrated or, contrary to tradition, intracellular-dehydrated rats, suggests that sodium appetite and thirst share more mechanisms than previously thought. Water deprivation has physiological and health effects in humans that might be related to the salt craving shown by our species. PMID- 17464435 TI - Matching salt intake to physiological need in rats using foraging protocols. AB - Several studies of the quantitative relationship between sodium need and sodium intake in rats are reviewed. Using acute diuretic treatment 24 h beforehand, intake matches need fairly accurately when intake is spread out in time by using a hypotonic solution of NaCl. In contrast, using a hypertonic solution, intake is typically double the need. Using the same diuretic treatment, although the natriuresis occurs within approximately 1 h, the appetite appears only slowly over 24 h. Increased plasma levels of aldosterone parallel the increased intake; however, treatment with metyrapone blocks the rise in aldosterone but has no effect on appetite. Satiation of sodium appetite was studied in rats using sodium loss induced by chronic diuretic treatment and daily salt consumption sessions. When a simulated foraging cost was imposed on NaCl access in the form of a progressive ratio lever press task, rats showed satiation for NaCl (break point) after consuming an amount close to their estimated deficit. The chronic diuretic regimen produced hypovolemia and large increases in plasma aldosterone concentration and renin activity. These parameters were reversed to or toward non depleted control values at the time of behavioral satiation in the progressive ratio protocol. Satiation mechanisms for sodium appetite thus do appear to exist. However, they do not operate quantitatively when concentrated salt is available at no effort, but instead allow overconsumption. There are reasons to believe that such a bias toward overconsumption may have been beneficial over evolutionary time, but such biasing for salt and other commodities is maladaptive in a resource-rich environment. PMID- 17464436 TI - Sodium taste threshold in children and its relationship to blood pressure. AB - Popular science has emphasized the risks of high sodium intake and many studies have confirmed that salt intake is closely related to hypertension. The present mini-review summarizes experiments about salt taste sensitivity and its relationship with blood pressure (BP) and other variables of clinical and familial relevance. Children and adolescents from control parents (N = 72) or with at least one essential hypertensive (EHT) parent (N = 51) were investigated. Maternal questionnaires on eating habits and vomiting episodes were collected. Offspring, anthropometric, BP, and salt taste sensitivity values were recorded and blood samples analyzed. Most mothers declared that they added "little salt" when cooking. Salt taste sensitivity was inversely correlated with systolic BP (SBP) in control youngsters (r = -0.33; P = 0.015). In the EHT group, SBP values were similar to control and a lower salt taste sensitivity threshold. Obese offspring of EHT parents showed higher SBP and C-reactive protein values but no differences in renin-angiotensin-aldosterone system activity. Salt taste sensitivity was correlated with SBP only in the non-obese EHT group (N = 41; r = 0.37; P = 0.02). Salt taste sensitivity was correlated with SBP in healthy, normotensive children and adolescents whose mothers reported significant vomiting during the first trimester (N = 18; r = -0.66; P < 0.005), but not in "non vomiter offspring" (N = 54; r = -0.18; nonsignificant). There is evidence for a linkage between high blood pressure, salt intake and sensitivity, perinatal environment and obesity, with potential physiopathological implications in humans. This relationship has not been studied comprehensively using homogeneous methods and therefore more research is needed in this field. PMID- 17464437 TI - Sex differences in angiotensin II- induced hypertension. AB - Sex differences in the development of hypertension and cardiovascular disease have been described in humans and in animal models. In this paper we will review some of our studies which have as their emphasis the examination of the role of sex differences and sex steroids in modulating the central actions of angiotensin II (ANG II) via interactions with free radicals and nitric oxide, generating pathways within brain circumventricular organs and in central sympathomodulatory systems. Our studies indicate that low-dose infusions of ANG II result in hypertension in wild-type male mice but not in intact wild-type females. Furthermore, we have demonstrated that ANG II-induced hypertension in males is blocked by central infusions of the androgen receptor antagonist, flutamide, and by central infusions of the superoxide dismutase mimetic, tempol. We have also found that, in comparison to females, males show greater levels of intracellular reactive oxygen species in circumventricular organ neurons following long-term ANG II infusions. In female mice, ovariectomy, central blockade of estrogen receptors or total knockout of estrogen a receptors augments the pressor effects of ANG II. Finally, in females but not in males, central blockade of nitric oxide synthase increases the pressor effects of ANG II. Taken together, these results suggest that sex differences and estrogen and testosterone play important roles in the development of ANG II-induced hypertension. PMID- 17464438 TI - Development of fetal nicotine and muscarinic receptors in utero. AB - The role of acetylcholine in the central and peripheral nervous systems is well established in adults. Cholinergic modulation of vascular functions and body fluid balance has been extensively studied. In the embryo-fetus, cholinergic receptors are widespread in the peripheral and central systems, including smooth muscle and the epithelial lining of the cardiovascular, digestive, and urinary systems, as well as in the brain. Fetal nicotine and muscarinic receptors develop in a pattern (e.g., amount and distribution) related to gestational periods. Cholinergic mechanisms have been found to be relatively intact and functional in the control of vascular homeostasis during fetal life in utero at least during the last third of gestation. This review focuses on the development of fetal nicotine and muscarinic receptors, and provides information indicating that central cholinergic systems are well developed in the control of fetal blood pressure and body fluid balance before birth. Therefore, the development of cholinergic systems in utero plays an important role in fetal vascular regulation, gastrointestinal motility, and urinary control. PMID- 17464440 TI - Bioaccumulation of metals in sediment elutriates and their effects on growth, condition index, and metallothionein contents in oyster larvae. AB - The bioavailability of Cd, Cu, Zn, and Pb from two metal-contaminated sediments (Bidassoa and Dunkerque) was studied using Crassostrea gigas larvae exposed to sediment elutriates. The metal contents within the sediments, the larvae and larval growth, the condition index, and the induction of metallothionein in the larvae were measured. The larval growth and condition index were only affected after exposure to the highest elutriates concentration (5 to 25%) from the most contaminated sediment (Dunkerque). Bioaccumulation of all metals was observed in larvae exposed to Dunkerque elutriatre; only Cu bioaccumulation was observed in the Bidassoa elutriate. The results from larvae exposed to both sediment elutriates show a strong correlation between bioaccumulated metal considered individually or in combination and the metallothionein level in larvae presenting no detrimental effect. On the other hand, in the case of larvae exposed to the highest Dunkerque elutriate concentration and showing the highest metal body burden, we observed a drop in the metallothionein level. These results indicate that metallothionein is a more sensitive indicator of heavy metal pollution than physiological endpoints taken into account in bioassays and could be proposed as an early biomarker of metal exposure in larvae. However, care must be taken with "fault control" due to the toxicological effect on larvae metabolism in the case of substantial contaminant exposure. PMID- 17464439 TI - The use of synthetic sub-urethral slings in the treatment of female stress urinary incontinence. AB - We set out to review the existing literature regarding the use of synthetic suburethral sling products for the treatment of female stress urinary incontinence. Products currently implanted are examined and scrutinized, and evidence regarding their efficacy and complication rates is noted. Additionally, specifics of presently utilized synthetic materials, including construction method and biocompatibility, are explored and directly correlated to currently marketed products. This investigation was undertaken with the use of the Medline database. Studies pertaining to synthetic or surgical mesh, as well as each specific suburethral sling product, are included. Our findings and ultimately our recommendations stem from the preponderance of evidence supporting the continued use of knitted, macroporous polypropelene mesh slings. Several existing marketed products detailed in the study fit this description. Specific reference is made to recent reports of vaginal erosions and deep space infections related to several specific products. A cautionary note is also made regarding the implantation of transobturator sling products currently marketed without the necessary pre-market testing, potentially placing the public at risk. From the currently available literature on biomaterials, it seems clear that knitted macroporous polypropylene is the material of choice for suburethral implantation. With respect to the means and techniques by which these mesh materials are suburethrally implanted, the surgeon's choice often dictates which method is used, but recent experience has demonstrated that the transobturator approach can be equally as effective as the traditional tension-free vaginal tape, with less associated morbidity. Ongoing randomized controlled trials will further clarify and distinguish between methods. PMID- 17464441 TI - Using a Sediment Quality Triad approach to evaluate benthic toxicity in the Lower Hackensack River, New Jersey. AB - A Sediment Quality Triad (SQT) study consisting of chemical characterization in sediment, sediment toxicity and bioaccumulation testing, and benthic community assessments was performed in the Lower Hackensack River, New Jersey. Chemistry data in sediment and porewater were evaluated based on the equilibrium partitioning approach and other published information to investigate the potential for chemical effects on benthic organisms and communities. Relationships were supported by laboratory toxicity and bioaccumulation experiments to characterize chemical effects and bioavailability. Benthic community results were evaluated using a regional, multimetric benthic index of biotic integrity and four heterogeneity indices. Evidence of slight benthic community impairment was observed in five of nine sediment sample stations. Severe lethal toxicity to amphipods (Leptocheirus plumulosus) occurred in four of these five stations. Although elevated total chromium concentrations in sediment (as high as 1900 mg/kg) were the rationale for conducting the investigation, toxicity was strongly associated with concentrations of polycyclic aromatic hydrocarbons (PAHs) rather than total chromium. PAH toxic units (SigmaPAH TU) in sediment and SigmaPAH concentrations in laboratory organisms from the bioaccumulation experiment showed a clear dose-response relationship with toxicity, with 0% survival observed in sediments in which SigmaPAH TU > 1-2 and SigmaPAH concentrations in Macoma nasuta were >2 micromol/g, lipid weight. Metals detected in sediment and porewater, with the possible exception of copper, did not correlate with either toxicity or levels in tissue, likely because acid volatile sulfide levels exceeded concentrations of simultaneous extracted metals at all sample locations. The study reinforces the value of using multiple lines of evidence approaches such as the SQT and the importance of augmenting chemical and biological analyses with modeling and/or other approaches to evaluate chemical bioavailability and toxicity of sediments. PMID- 17464442 TI - Fluctuating asymmetry in insecticide-resistant and insecticide-susceptible strains of the maize weevil, Sitophilus zeamais (Coleoptera: Curculionidae). AB - Insecticide resistance is an evolutionary response of specific insect populations subjected to exposure and consequent selection by an insecticide. As such, this phenomenon is important as a biomonitoring strategy and also has economical importance in the case of insect-pests by compromising their control. Fluctuating asymmetry (FA) of bilateral traits is a measure of developmental instability also suggested as a monitoring tool for environmental pollution with potential consequences for fitness. Responses to selective agents might have pleiotropic effects influencing development and phenotype, which has yet to be examined for agricultural insecticides and insect-pests. Higher levels of FA are expected in the insecticide-resistant strains, which are usually at a selective disadvantage, relative to susceptible strains, in the absence of the insecticide. Two insecticide-resistant strains and an insecticide-susceptible strain of the maize pest insect Sitophilus zeamais (Coleoptera: Curculionidae) were subjected to FA measurements in 12 traits of wing veins and tibias of 100 individuals of both sexes of each strain. The insecticide-resistant strains showed lower FA than the susceptible strain, in contrast with the initial expectation. An extended period of insecticide selection probably led to the evolution of fitness-modifier genes improving the performance of the resistant genotypes, reducing their FA levels, and leading to their eventual fixation in the population. In addition, one insecticide-resistant strain and the insecticide-susceptible strain showed significant differences in FA between sexes, with more symmetrical males suggesting possible sexual selection by the females. The observed results have potential consequences for insecticide-resistance evolution and dispersal. PMID- 17464443 TI - Selenium and metal concentrations in waterbird eggs and chicks at Agassiz National Wildlife Refuge, Minnesota. AB - Exceptionally high cadmium (Cd) and chromium (Cr) concentrations were reported in eggs, feathers, or livers of selected waterbird species nesting at Agassiz National Wildlife Refuge (Agassiz) in 1994. Ten- to 15-day-old Franklin's gull (Larus pipixcan), black-crowned night-heron (Nycticorax nycticorax), and eared grebe (Podiceps nigricollis) chicks were collected in 1998, 1999, and 2001 at Agassiz and analyzed for selenium (Se) and metals including Cd and Cr. Freshly laid eggs were collected in 2001 from Franklin's gull, black-crowned night-heron, eared grebe, and pied-billed grebe (Podilymbus podiceps) nests at Agassiz. Based on a multivariate analysis, the pattern of Se and metal concentrations differed among species for eggs, chick feathers, and chick livers. Low Cd and Cr concentrations were measured in eggs, chick livers, and chick feathers of all four species. Mercury concentrations in black-crowned night-heron and eared grebe eggs collected from Agassiz in 2001 were lower than concentrations reported in 1994. Se and metal concentrations, including Cd and Cr, in waterbird eggs and chicks collected at Agassiz in 1998, 1999, and 2001 were not at toxic levels. PMID- 17464444 TI - Immune status, carotenoid coloration, and wing feather growth in relation to organochlorine pollutants in great black-backed gulls. AB - Previous ecotoxicological studies have documented relationships between residues of various organochlorines (OCs) and immune status, carotenoid colors, and wing feather growth in different bird species. In this study, the density of white blood cells (WBC), carotenoid colors, and length of the same feathers on each wing were measured in breeding great black-backed gulls (Larus marinus) on the coast of northern Norway, and related to the blood residues of five OCs, including HCB (hexachlorobenzene), beta-HCH (beta-hexachlorocyclohexane), p,p' DDE (p, p'-dichlorodiphenyldichloroethylene), oxychlordane, and polychlorinated biphenyl (PCB), in addition to SigmaOC. Neither, WBC density nor carotenoid colors were significantly related to blood residues of any of the OCs, suggesting that OC levels may have been too low to significantly affect these outcome parameters. However, in the colony where the OC concentrations were highest, there was a weak but significantly positive relationship between the probability of having different length of feathers on each wing and levels of PCB and SigmaOC, in males. Thus varying length of the wing primaries may reflect adverse impacts of OCs in great black-backed gulls. However, in gulls with moderate levels of OCs, it is probably not a sensitive indicator of progressing ecological impacts of OCs, since such adverse ecological relationships were found in the breeding colonies where there were no relationships between differences in wing feather lengths and OCs. PMID- 17464445 TI - Testicular teratocarcinoma associated with testicular microlithiasis. PMID- 17464446 TI - Preferential response of glutathione-related enzymes to folate-dependent changes in the redox state of rat liver. AB - BACKGROUND: Oxidative stress likely constitutes an important contributing factor in the onset of degenerative diseases associated with folate deficiency. Direct, as well as homocysteine-linked, antioxidant properties of folate could explain its preventive effect on these pathologies. AIM OF THE STUDY: Our study aimed at determining the changes in the redox status of adult rats as a function of folate intake. METHODS: Adult male rats were pair-fed for 4 weeks with a semi-synthetic diet containing 0, 0.5, 1.5, 8 or 20 mg of folic acid/kg. Folate and homocysteine concentrations, redox status markers and antioxidant enzyme activities were measured in the plasma and/or liver of the rats. A principal component analysis of the overall data was performed to draw a general scheme of the changes observed between the conditions. RESULTS: Folate deficiency caused increased homocysteinemia and features of oxidative stress including reduced plasma antioxidant capacity together with increased lipid peroxidation in liver and heart. This was associated with an increase in the specific activity of several enzymes involved in liver glutathione metabolism (glutathione peroxidase, glutathione reductase and glutathione S-transferase), suggesting an adaptive tissue response to the oxidative stress induced by folate deficiency. In contrast, no such variation was observed for hepatic superoxide dismutase and catalase. CONCLUSION: Despite no changes in hepatic levels of total glutathione, our findings indicate that glutathione-dependent antioxidant pathways could be particularly involved in the compensatory mechanism committed by liver to counteract the oxidative stress induced by folate deficiency. They also suggest that folate supplementation may not be associated with a better antioxidant protection of rats. PMID- 17464447 TI - What leads from dead-end? AB - The 129 mouse strain develops congenital testicular germ cell tumors (TGCTs) at a low frequency. TGCTs in mice resemble the testicular tumors (teratomas) that occur in human infants. The genes that cause these tumors in 129 have not been identified. The defect at the Ter locus increases TGCT incidence such that 94% of 129-Ter/Ter males develop TGCTs. The primary effect of the Ter mutation is progressive loss of primordial germ cells (PGCs) during embryonic development. This results in sterility in adult Ter/Ter mice on all mouse strain backgrounds. However, on the 129 background, Ter causes tumor development in addition to sterility. Therefore, Ter acts as a modifier of 129-derived TGCT susceptibility genes. Ter was identified to be a mutation that inactivates the Dead-end1 (Dnd1) gene. In this perspective, I discuss the possible areas of future investigations to elucidate the mechanism of TGCT development due to Dnd1 inactivation. PMID- 17464448 TI - Using blood brain barrier disruption by methamphetamine for drug delivery. PMID- 17464449 TI - Hepatocyte growth factor production is stimulated by gangliosides and TGF-beta isoforms in human glioma cells. AB - Hepatocyte growth factor (HGF) is a pleiotrophic cytokine that stimulates motility and invasion of several cancer cell types and induces angiogenesis, which is known to be expressed in several malignancies including glioma. The effect of transforming growth factor-beta (TGF-beta) isoforrns as well as gangliosides on HGF production was investigated in human glioma cell lines. TGF beta isoforms and gangliosides were found to differentially stimulate HGF production by these cells. The ganglioside GD3 enhanced this release to the greatest extent and the stimulation was more marked in a glioblastoma cell line than in the two other anaplastic astrocytoma cell lines. These results suggest that both TGF-betas and gangliosides may act as indirect angiogenic factors by stimulating HGF secretion. PMID- 17464450 TI - Always a surprise, even a wonder: poetry and commentary. PMID- 17464451 TI - "Compliance" to "concordance": a critical view. AB - Advocates of "concordance" describe it as a new model of shared decision-making between physicians and patients based on a partnership of equals. "Concordance" is meant to make obsolete the notion of "compliance," in which patients are seen as, ideally, following doctors' orders. This essay offers a critical view of concordance, arguing that the literature itself on concordance, including materials at the web site of Medicines Partnership, the implementation arm in Great Britain of the concordance model, is full of contradiction; concordance, in fact, harbors an ideology of compliance. The essay suggests that an improvement in patient medication use will more likely come from a frank consideration of the relation of compliance issues and commercial ones, and that a key question across domains is, "how are patients/health agents/consumers persuaded to acquire certain drugs and take them as directed?" PMID- 17464452 TI - Immunohistochemical study of the presence of mast cells in idiopathic orbital inflammatory pseudotumor: possible role of mast cells in the course of its pathogenesis. AB - BACKGROUND: It is well known that mast cells (MC) are involved in fibrosis and many forms of chronic inflammation. Chronic inflammatory cells infiltration and fibrosis in various orbital tissues are the main histopathologic features in patients with idiopathic orbital inflammatory pseudotumor (IOIP). Whether MC is involved in the course of chronic inflammatory conditions of IOIP is not yet clear. We sought to investigate the distribution of MCs in samples of IOIP and to explore the possible role of MC in the course of its pathegenesis. METHODS: Immunohistochemistry with tryptase monoclonal antibody (a specific mast-cell surface marker) was used in 53 different subtypes of IOIP specimens including 19 of the lymphocyte infiltrative type, 22 of the fibrotic type, 12 of the mixed type and 4 specimens of normal orbital tissue. The number of positive stained MC was counted by light microscopy. The differences of the number of MC between various subtypes of IOIP were analyzed. RESULTS: The average number of positive stained MC in the normal control group was 33.33 +/- 4.72 /mm(3), whereas the average numbers of positive stained MC in the lymphocyte infiltrative subtype group, the mixed subtype group and the fibrotic subtype group were 306.35 +/- 55.81 /mm(3), 662.93 +/- 115.28 /mm(3) and 813.44 +/- 146.56 /mm(3), respectively. Compared with the normal control, the number of MC increased significantly in all three subtypes of IOIP samples (P < 0.01). The number of MC in fibrotic subtype IOIP was the largest, followed by mixed subtype IOIP, and the lymphocyte infiltrative subtype IOIP (P < 0.05). The MC were distributed mainly around small vessels and in collagen fibers. CONCLUSION: Mast cells may play an underappreciated role in the fibrosis of IOIP. PMID- 17464453 TI - Enzymatic lysis of microbial cells. AB - Cell wall lytic enzymes are valuable tools for the biotechnologist, with many applications in medicine, the food industry, and agriculture, and for recovering of intracellular products from yeast or bacteria. The diversity of potential applications has conducted to the development of lytic enzyme systems with specific characteristics, suitable for satisfying the requirements of each particular application. Since the first time the lytic enzyme of excellence, lysozyme, was discovered, many investigations have contributed to the understanding of the action mechanisms and other basic aspects of these interesting enzymes. Today, recombinant production and protein engineering have improved and expanded the area of potential applications. In this review, some of the recent advances in specific enzyme systems for bacteria and yeast cells rupture and other applications are examined. Emphasis is focused in biotechnological aspects of these enzymes. PMID- 17464454 TI - Metabolic syndrome and gastrointestinal diseases. AB - Metabolic syndrome is a cluster of metabolic abnormalities consisting essentially of obesity, especially abdominal obesity. Metabolic syndrome has been highlighted as a risk factor for cardiovascular and other chronic diseases. Obesity has been implicated in various gastrointestinal diseases such as gastroesophageal reflux diseases and colorectal cancer. Recently, abdominal obesity has been shown to be more important than obesity as expressed by an elevated body mass index as a causative factor for the development of these diseases. In addition to the mechanical effects of obesity, such as an increase in intra-abdominal pressure from large amounts of adipose tissue, substances that adipose tissues secrete, such as tumor necrosis factor-alpha, interleukin-6, leptin, and insulin-like growth factor-1, have been proposed to be pathogenic links to these diseases. In this review, we discuss the association of metabolic syndrome or the individual components of metabolic syndrome, focusing on obesity and abdominal obesity, with gastrointestinal diseases. PMID- 17464455 TI - Does long-term medication with a proton pump inhibitor induce a tolerance to H2 receptor antagonist? AB - BACKGROUND: Previous reports suggest that complete tolerance to H2 receptor antagonists (H2RAs) in patients with regular H2RA medication may be due to hypergastrinemia-increased histamine synthesis or upregulation of H2 receptors. As proton pump inhibitors (PPIs) have been reported to induce hypergastrinemia (similar to H2RAs), patients receiving long-term medication with PPIs may show tolerance to preanesthetic H2RA. Therefore, we studied the efficacy of an H2RA, roxatidine, in patients receiving long-term PPI medication. METHODS: Effects of H2RA in 15 surgical patients receiving a regular PPI for more than 4 weeks (PPI+H2RA group) were compared with those in 30 patients not receiving regular PPIs or H2RAs (None+H2RA group and None+None group, n = 15 each). Oral roxatidine was given to both PPI+H2RA and None+H2RA group patients as an anesthetic premedication, while it was not given to None+None group patients. Gastric volume and pH were measured after induction of anesthesia. RESULTS: Gastric pH and volume (ml) in the PPI+H2RA group (5.79 +/- 1.61 and 9.1 +/- 16.7, respectively) were both similar to those in the None+H2RA group (5.54 +/- 2.20 and 9.7 +/- 10.8, respectively) but were significantly higher (gastric pH) and lower (volume) than in the None+None group (2.29 +/- 1.84 and 29.3 +/- 22.8, respectively, P < 0.01). CONCLUSIONS: These data suggest that long-term PPI medication may not induce a tolerance to H2RAs. PMID- 17464456 TI - Fecal pellet output does not always correlate with colonic transit in response to restraint stress and corticotropin-releasing factor in rats. AB - BACKGROUND: Fecal pellet output has been assessed as a colonic motor activity because of its simplicity. However, it remains unclear whether an acceleration of colonic transit correlates well with an increase in fecal pellet output. We examined the causal relationship between colonic transit and fecal pellet output stimulated by the central application of corticotropin-releasing factor (CRF) and restraint stress. METHODS: Immediately after intracisternal injection of CRF, (51)Cr was injected via a catheter positioned in the proximal colon. Ninety minutes after (51)Cr injection, the total number of excreted feces was counted, and then the rats were killed. The radioactivity of each colonic segment was evaluated, and the geometric center (GC) of the distribution of (51)Cr was calculated. For the restraint stress study, after administration of (51)Cr into the proximal colon, rats were submitted to wrapping restraint stress for 90 min. Then they were killed, and GC was calculated. RESULTS: Both restraint stress and CRF significantly accelerated colonic transit. There was a positive correlation observed between fecal pellet output and GC of colonic transit in response to restraint stress, but not CRF, when the number of excreted feces was more than three. In contrast, there was no significant correlation observed between the two in stress and CRF when the number of excreted feces was less than two. CONCLUSIONS: The acceleration of colonic transit in response to restraint stress and central administration of CRF does not always correlate with an increase in fecal pellet output. PMID- 17464457 TI - Endoscopic stenting versus surgical gastroenterostomy for palliation of malignant gastroduodenal obstruction: a meta-analysis. AB - BACKGROUND: We attempted to elucidate the current status of endoscopic self expanding metal stents for palliation of malignant gastroduodenal obstruction in comparison with surgical gastroenterostomy. METHODS: Original articles and abstracts published from January 1990 to September 2006 were searched in Medline, EMBASE, and Cochrane Controlled Trials Register databases. Clinical appraisal and data extraction were independently conducted by two reviewers. Statistical analysis was performed by meta-analysis using a random effects model. Weighted mean differences with 95% confidence intervals (CI) were used to analyze continuous variables. Odds ratios with 95% CI were calculated for dichotomous variables. RESULTS: The outcomes of 307 procedures from nine studies were analyzed. Endoscopic stenting was found to be associated with higher clinical success (P = 0.007), a shorter time from the procedure to starting oral intake (P < 0.001), less morbidity (P = 0.02), lower incidence of delayed gastric emptying (P = 0.002), and a shorter hospital stay (P < 0.001) than surgical gastroenterostomy. There was no significant difference between the two groups in the analysis of 30-day mortality. CONCLUSIONS: Endoscopic stenting may be a feasible alternative to surgery for the palliation of inoperable malignant gastroduodenal obstruction, with a high clinical success and low morbidity rate. Additional well-designed randomized controlled trials with larger sample sizes are expected to further reinforce this conclusion. PMID- 17464458 TI - Pancreatic diabetes in a follow-up survey of chronic pancreatitis in Japan. AB - BACKGROUND: We aimed to determine the cumulative rate of diabetes mellitus (DM) and the risk factors for DM in patients with chronic pancreatitis (CP) in Japan. METHODS: We conducted a follow-up survey of CP in 2002 in patients registered as having CP in 1994, and confirmed 656 patients to be checked in regard to the survey items concerning diabetes. We analyzed the cumulative rate of DM and the risk factors for DM over an 8-year follow up period. RESULTS: In 1994, 35.1% of 656 CP patients had DM, and the incidence of diabetes had increased to 50.4% in 2002. Of 418 patients without diabetes in 1994, 28.9% (121/418) were newly diagnosed with DM in 2002. Alcoholic CP was the most common type of CP in patients with newly developed diabetes, accounting for 67.8%. The incidence of DM was highest in those with alcoholic CP (34.3%) followed by idiopathic CP (23.0%). The risk of diabetes increased 1.32-fold after the onset of pancreatic calcification. Of 121 patients with newly diagnosed DM in 2002, 37 (30.6%) had pancreatic stones in 1994 and 49 (40.5%) had a stone in 2002. The highest incidence of newly diagnosed DM was observed in patients with continuous alcoholic intake (40.9%). Patients treated with camostat mesilate developed DM less frequently than those without camostat mesilate. CONCLUSIONS: The present study showed that the incidence of DM in patients with CP increased with time. Of 418 CP patients without DM in 1994, 28.9% developed DM over a period of 8 years. Continuous alcoholic intake aggravated CP and increased the risk of DM in those with CP. PMID- 17464459 TI - A weak association between occult HBV infection and non-B non-C hepatocellular carcinoma in Japan. AB - BACKGROUND: In Japan, approximately 10% of hepatocellular carcinoma (HCC) patients are negative for both hepatitis B surface antigen (HBsAg) and antibodies to hepatitis C virus (anti-HCV), i.e., they constitute the so-called category of non-B non-C (NBNC) HCC. Little is known about the characteristics of NBNC-HCC. METHODS: Potential risk factors for carcinogenesis (including occult HBV infection [HBsAg is negative but HBV DNA is positive by polymerase chain reaction (PCR)], obesity, and diabetes) were assessed in 233 HCC patients grouped according to hepatitis virus serological status (152 with HCV-HCC, 36 with HBV HCC, and 45 with NBNC-HCC). RESULTS: The prevalence of patients with obesity or diabetes was significantly higher in the NBNC-HCC group than in the HBV-HCC group. The same trend was observed even when patients with massive alcohol intake were excluded from the analysis. Only 8 patients (18%) in the NBNC-HCC group had detectable serum HBV DNA, and this was at very low levels (HBV/Ce/C2 and HBV/D were determined in 7 and 1 patients, respectively). In the NBNC-HCC group, the determined nucleotide sequences of the enhancer II/core promoter/precore/core region did not contain any HCC-associated mutations, whereas 25 of 30 patients in the HBV-HCC group carried strains with C1653T, T1753V, and/or A1762T/G1764A mutations. CONCLUSIONS: A weak association between occult HBV infection and HCC development was observed in the NBNC patients. This study indicates that nonalcoholic steato-hepatitis should be further investigated to assess its contribution to HCC development in this category of patients. PMID- 17464460 TI - Percutaneous ultrasound-guided radiofrequency ablation of hepatocellular carcinoma with artificially induced pleural effusion and ascites. AB - BACKGROUND: Ultrasound-guided procedures are sometimes of limited use because the tumor is located under the diaphragm or near the surface of the liver. We investigated the safety and efficacy of radiofrequency ablation (RFA) with artificial pleural effusion and/or artificial ascites. METHODS: Between January 2002 and May 2006, 43 lesions in 36 patients with hepatocellular carcinoma (HCC) were treated by RFA with artificial pleural effusion and/or artificial ascites. RESULTS: Artificial pleural effusion allowed visualization of the whole tumor for 36 (83.7%) of the 43 lesions that were otherwise not detectable or poorly visible. Artificial ascites was also helpful in visualizing whole tumors that could not be visualized with only artificial pleural effusion. In all lesions, artificial pleural effusion and/or artificial ascites were helpful in performing percutaneous RFA. Artificial ascites was useful for creating a space between the liver's surface and the skin or diaphragm to avoid burns. Adverse effects after the induction of artificial pleural effusion included pneumonia in one patient and temporary atelectasis in another patient. Severe side effects were not observed. Complete necrosis after RFA was obtained in 43 (100%) of the 43 lesions. During a mean follow-up period of 31.8 +/- 5.8 months, local recurrence at the ablation site was found in none of the 43 lesions. CONCLUSIONS: Percutaneous RFA with artificial pleural effusion and/or artificial ascites was a safe and useful treatment that resulted in good local control of HCC. PMID- 17464461 TI - Liver steatosis, but not fibrosis, is associated with insulin resistance in nonalcoholic fatty liver disease. AB - BACKGROUND: To address the hypothesis that liver steatosis causes systemic insulin resistance, we sought to determine the liver histological feature that most strongly contributes to insulin resistance in patients with nonalcoholic fatty liver disease (NAFLD). METHODS: Liver biopsy specimens were obtained from 131 patients with clinically suspected NAFLD. The stage, grade of nonalcoholic steatohepatitis (NASH), and level of steatosis were scored and analyzed in relation to the homeostasis model assessment of insulin resistance (HOMA-IR) and the metabolic clearance rate (MCR), measured using the glucose clamp method. RESULTS: In the univariate analysis, the degree of hepatic steatosis (r = 0.458, P < 0.001), stage (r = 0.360, P < 0.001), and grade (r = 0.349, P < 0.01) of NASH were significantly correlated with the HOMA-IR. Multiple regression analysis adjusting for age, sex, body mass index, and each histological score showed that steatosis was significantly and independently associated with HOMA-IR (coefficient = 1.42, P < 0.001), but not with the stage (coefficient = 0.33, P = 0.307) or grade (coefficient = 0.67, P = 0.134) of NASH. Similar independent relationships were observed between steatosis and MCR, but the relationship was weaker (coefficient = -0.98, P = 0.076). CONCLUSIONS: Steatosis of the liver, but not the stage or the grade of NASH, is associated with insulin resistance in patients with NAFLD. PMID- 17464462 TI - Choledochocele associated with superficial spreading cancer with cholesterolosis of the bile duct. AB - A 74-year-old woman, who had suffered from acute cholangitis, was referred to our department for further evaluation of the biliary tree. A diagnosis of choledochocele was made by endoscopic ultrasonography and endoscopic retrograde cholangiopancreatography. A small flat elevation with cholesterolosis was observed in the lower bile duct, and circumferential wall thickening was apparent in some parts of the upper and middle bile duct by intraductal ultrasonography and peroral cholangioscopy. With the diagnosis of choledochocele associated with superficial spreading cancer and cholesterolosis of the bile duct, pylorus preserving pancreaticoduodenectomy was carried out. Microscopically, the tumor had spread extensively from the lower to the upper bile duct. Its invasion was limited to the mucosa for the most part, with microinvasion of the subserosa in the upper bile duct. In the lower bile duct, foamy cells were located beneath the neoplastic epithelium of the elevation. The number of cases of choledochocele associated with biliary cancer is becoming higher than previously reported. This anomaly may play some role in the development of biliary malignancy. Histological examination seems to be mandatory before making a diagnosis of cholesterolosis of the bile duct, since this condition may occasionally accompany cancer. PMID- 17464463 TI - Four-year follow-up of the first case of gastroesophageal reflux disease treated with endoluminal gastroplication in Japan. PMID- 17464464 TI - Influence of phlebotomy on iron-related gene expression levels in the livers of patients with chronic hepatitis C. PMID- 17464465 TI - On the relationship between intracerebral venous pressure, intracranial pressure and brain edema. PMID- 17464468 TI - Neurodegeneration and Alzheimer's disease: the lesson from tauopathies. AB - The Amyloid Cascade Hypothesis suggests that the decisive event in Alzheimer's disease (AD) is the deposition of fibrils of beta-amyloid protein (Abeta). The main objection to this hypothesis is the weak correlation between plaque load and severity of dementia. The good correlation between synaptic loss and dementia suggests that AD may be regarded as a synaptic failure. The toxicity of Abeta depends on its state of aggregation. The most important implication derived from the studies of tau gene mutations in a familial form of frontotemporal dementia (FTDP-17) is that the mutation itself is sufficient to cause neuronal loss. Several recent data suggest that apoptotic mechanisms may represent the missing link between Abeta deposition and proteolysis of tau, an early event in the pathogenic sequence of AD. Collectively, these observations suggest a model of AD whereby overproduction or reduced clearance of Abeta initiates a cascade of events that lead to neuronal loss directly or through post-translational modification of tau. PMID- 17464469 TI - Panayiotopoulos syndrome: diagnosis and management. AB - Panayiotopoulos syndrome is a relatively frequent and benign epileptic syndrome, characterised by predominantly autonomic symptoms and/or simple motor focal seizures followed or not by impairment of consciousness. Interictal EEG shows occipital spikes although multifocal spikes with high amplitude sharp-slow wave complexes at various locations can be present. This syndrome can imitate gastroenteritis, encephalitis, syncope, migraine, sleep disorders or metabolic diseases. The peculiar aspects should be known not only by epileptologists but also by general doctors because a correct diagnosis would avoid aggressive interventions and concerns on account of its benign outcome. This review focuses on the main clinical and EEG features of this epilepsy underlining its typical and atypical symptoms and its management. PMID- 17464470 TI - The Frontal Behavioural Inventory (Italian version) differentiates frontotemporal lobar degeneration variants from Alzheimer's disease. AB - The objective was to evaluate the construct validity of the Italian version of the Frontal Behavioural Inventory (FBI) and its usefulness in the differential diagnosis of dementias. Standard criteria were used in the clinical diagnosis of dementias in 83 patients and 33 agematched healthy volunteers. The FBI scale was translated from English into Italian language and back-translated. Cronbach's alpha, inter-rater and test-retest reliability, FBI convergent validity and discriminant analysis were calculated. FBI profile was compared between patients affected by frontotemporal lobar degeneration (FTLD) and Alzheimer's disease (AD). The FBI showed a high internal consistency and inter-rater reliability and it distinguished normal behavioural conditions from those presented in FTLD or AD. An 86.8% diagnostic accuracy was calculated by the discriminant analysis, selecting only age at disease onset and FBI, and particularly distinguishing behavioural variants within the FTLD spectrum. FTLD patients showed a characteristic behavioural profile. The FBI might be a reliable and useful diagnostic tool for dementias in clinical practice. PMID- 17464471 TI - Imaging of cerebrospinal fluid space and movement of hydrocephalus mice using near infrared fluorescence. AB - We have studied optical imaging of mice cerebrospinal fluid (CSF) space and flow using near infrared (NIR) fluorescence. We applied our method to image CSF space and flow in a mice hydrocephalus model. Hydrocephalus was induced in mice with intracranial injections of transforming growth factor. Hydrocephalic and control mice were imaged using our NIR fluorescence imaging system. Hydrocephalic mice showed diminished intracranial CSF flow. Our system is sufficient to show altered CSF flow in a mouse hydrocephalus model. Optical imaging using near infrared is an effective modality to image CSF space and movement. PMID- 17464472 TI - Spontaneous neuronal activity of the posterior hypothalamus in trigeminal autonomic cephalalgias. AB - Microrecordings of three neurons were obtained at the target site in three patients with trigeminal autonomic cephalalgias who were implanted with deep brain stimulators in the posterior hypothalamus. Two patients had chronic cluster headache, one short unilateral neuralgiform headache with conjunctival injection and tearing. Average firing rate was around 24 spikes/s. All neurons were firing randomly, and for most of the recordings in tonic fashion. In one patient, tactile stimulation of the ophthalmic branch, contralateral to the recording site, decreased the firing rate. Neuronal activity in these patients was similar to that reported in animal studies of the posterior hypothalamus. Positioning deep brain stimulators in the posterior hypothalamus may offer a tool to better characterise the activity of this part of the brain in humans. PMID- 17464473 TI - Hallucinations and sleep-wake cycle in Alzheimer's disease: a questionnaire-based study in 218 patients. AB - The aim was to evaluate the relationship between hallucinations and the sleep wake cycle in a sample of Alzheimer's disease (AD) patients in the early-moderate stage. Two hundred and eighteen AD patients (66 males, 152 females, mean age 74.3+/-6.85) were administered a sleep questionnaire in the presence of a care giver. Twenty-six out of 218 (12%) reported the occurrence of hallucinations, mainly visual. In 18/28 (69%) hallucinations occurred when the patient was awake and in 8 (31%) hallucinations were reported to occur close to a specific phase of the sleep-wake cycle. Vivid dreams were reported in 25/218 (11%) and violent sleep-related and dream-related behaviours (probable REM behaviour episodes) in 22/218 (10%). Both REM phenomena were more frequent in AD hallucinators than in AD non-hallucinators (26.9% vs. 9.3%, and 26.9% vs. 7.8%, p<0.007). Our data indicate a lower incidence of hallucinations and presumable REM behaviour disorder (RBD) in AD, at least in the early-moderate phase, than that observed in synucleinopathies. However, the higher occurrence of vivid dreams and RBD in AD patients with hallucinations compared to those without hallucinations indicates a potential role of disordered REM sleep in influencing the occurrence of hallucinations in AD, similar to what has been observed in synucleinopathies. PMID- 17464474 TI - Gait patterns of a patient with myoclonus of a lower limb, when OFF and ON treatment with antiepileptic drugs. AB - We describe kinematics, kinetics and electromyographic patterns of a patient with spinal myoclonus of the left lower limb, during walking. Gait analysis was performed when the patient was OFF and ON his treatment with antiepileptic drugs. When OFF, we mainly observed clonic bursts and out-of-phase activations of m. tibialis anterior and m. rectus femoris, with increased hip flexion, reduction of knee flexion during swing and excessive ankle dorsal flexion. Furthermore, large oscillations of knee moment of force and power during stance phase were also observed. These abnormal patterns markedly recovered when ON drugs. PMID- 17464475 TI - Dropped head as an unusual presenting sign of myasthenia gravis. AB - Prominent or isolated weakness of cervical extensor muscles is a relatively rare clinical sign. Commonly, this is known as "dropped-head syndrome". This abnormal flexion of the head may occur in a variety of neuromuscular diseases and in a few non-neurological disorders as well. The case we describe concerns a 61-year-old woman with dropped-head syndrome as the unique complaint of myasthenia gravis. PMID- 17464476 TI - Progressive topographical disorientation: a case of focal Alzheimer's disease. AB - We describe a follow-up study of a patient with a selective, progressive impairment of topographical orientation. The patient's topographical difficulties were evident only in unfamiliar surroundings at the beginning of the observation period but later on they were observed even at home. Serial neuropsychological tests demonstrated a progressive impairment of visuospatial abilities with sparing of the other cognitive domains; only at the last assessment, about six years after early disturbances and three years after the first evaluation, the patient developed the typical cognitive impairments of Alzheimer's disease (AD). This case represents a focal variant of AD not previously described and suggests that the neuronal pathways underlying spatial orientation may be selectively damaged by the degenerative process. PMID- 17464477 TI - Ischaemic stroke, factor V Leiden heterozygosity and left atrial thrombosis in sinus rhythm: a case report. AB - We describe a 63-year-old man in sinus rhythm (SR) with an ischaemic stroke involving basal ganglia region on the right side. The patient was known to be heterozygous for factor V Leiden (FVL) mutation. On diagnostic work-up, no arterial sources of embolism were found. Transoesophageal echocardiography evidenced a left atrial (LA) thrombosis without relevant cardiopathies. LA thrombosis is generally associated to atrial fibrillation, atrial enlargement, mitral valve stenosis and left ventricular dysfunction, whereas mitral regurgitation is considered protective. To our knowledge, this is the first report of cardioembolic stroke related to a LA thrombosis in a patient in SR without risk factors for thrombus formation except for FVL heterozygosity. PMID- 17464478 TI - Concurrence of non-myasthenic symptoms with myasthenia gravis. AB - Myasthenia gravis (MG) is a disease that is known to be accompanied by various complications. But the relationship between these complications and MG and the treatment for these complications still partly remain unknown. We report two cases of MG with unusual complications. The first one is a case of a 72-year-old woman with lingual dyskinesia, and the second is a 28-year-old man with dysgeusia. Both symptoms improved in parallel after the treatment of MG. Here we report these cases and review similar cases in the literature. PMID- 17464479 TI - Basilar artery fenestration in a woman with basilar-type migraine. PMID- 17464482 TI - Revision of Potamotrygonocotyle Mayes, Brooks & Thorson, 1981 (Platyhelminthes: Monogenoidea: Monocotylidae), with descriptions of four new species from the gills of the freshwater stingrays Potamotrygon spp. (Rajiformes: Potamotrygonidae) from the La Plata river basin. AB - The only known monocotylid genus to parasitise Neotropical freshwater stingrays (Potamotrygonidae) is Potamotrygonocotyle Mayes, Brooks & Thorson, 1981, a monotypic genus erected to accommodate P. tsalickisi Mayes, Brooks & Thorson, 1981. For more than 20 years, no other species has been recognised in this genus, but new efforts to survey the diversity of parasites inhabiting potamotrygonids have revealed the existence of new species and the need to redefine the genus. Here, the generic diagnosis of Potamotrygonocotyle is amended, P. tsalickisi is redescribed and four new species are recognised and described based on samples collected from the gills of freshwater potamotrygonids from the La Plata river basin: Potamotrygonocotyle chisholmae n. sp. and P. dromedarius n. sp. from Potamotrygon motoro; Potamotrygonocotyle eurypotamoxenus n. sp. from Potamotrygon cf. motoro (type-host), P. castexi, P. falkneri and P. histrix; and Potamotrygonocotyle uruguayensis n. sp. from Potamotrygon brachyura. Potamotrygonocotyle is characterised by species possessing: (1) slightly sinuous sclerotised ridges on all septa; (2) two pairs of the dorsal haptoral accessory structures associated with the four posterior peripheral loculi and with anterior dorsal haptoral accessory structure bilobate or semicircular; and (3) male copulatory organ without an accessory piece. PMID- 17464481 TI - Fecal steroid monitoring for assessing gonadal and adrenal activity in the golden eagle and peregrine falcon. AB - We examined the efficacy of noninvasive monitoring of endocrine function via fecal steroid immunoassays in the golden eagle and peregrine falcon. High pressure liquid chromatography analyses of fecal glucocorticoid metabolites (fGCM) revealed that minor percentages of immunoreactive fGCM co-eluted with [(3)H]corticosterone in both sexes of the eagle (2.5-2.7%) and falcon (7.5 11.9%). In contrast, most fecal estrogen metabolites in eagle and falcon females co-eluted with radiolabeled estradiol-17beta ([(3)H]; 57.6, 64.6%, respectively) or estrone ([(3)H]; 26.9, 4.1%, respectively). Most fecal progestin metabolite immunoreactivity in the female eagle (24.8%) and falcon (21.7%) co-eluted with progesterone ([(14)C]). Most fecal androgen metabolite immunoreactivity in eagle (55.8%) and falcon (63.7%) males co-eluted with testosterone ([(14)C]). Exogenous adrenocorticotropin hormone induced increased fGCM excretion above pre-treatment in both species, but only significantly (P < 0.05) in the eagle. Both species showed increased fGCM after saline administration, suggesting the detection of 'handling stress.' Both species exhibited enterohepatic and renal recirculation of administered steroids as demonstrated by biphasic and triphasic excretion patterns. Thus, noninvasive fecal hormone monitoring is a valid and promising tool for assessing gonadal and adrenal status in rare and threatened birds-of prey. PMID- 17464483 TI - A revision of Megalonchos Baer & Euzet, 1962 (Tetraphyllidea: Onchobothriidae), with the description of two new species and transfer of two species to Biloculuncus Nasin, Caira & Euzet, 1997. AB - The onchobothriid tapeworm genus Megalonchos Baer & Euzet, 1962 is revised and the generic diagnosis amended based on the examination of some of Southwell's material of M. mandleyi (Southwell, 1927) Baer & Euzet, 1962, the type-specimens of M. dubius Prudhoe, 1969 and M. musteli Prudhoe, 1969, and material of two new species, M. sumansinghai n. sp. and M. shawae n. sp., collected from the snaggletooth shark Hemipristis elongatus off northern Australia. Based on their possession of two pairs of uni-pronged hooks (rather than one pair of bi-pronged hooks) and possession of, rather than lack of, post-vaginal testes, M. dubius and M. musteli are transferred to Biloculuncus Nasin, Caira & Euzet, 1997 as B. dubius (Prudhoe, 1969) n. comb. and B. musteli (Prudhoe, 1969) n. comb. Both new species of Megalonchos differ from M. mandleyi in their possession of conspicuously smaller hooks and shorter cephalic peduncles. The new species are readily distinguished from one another in that, whereas the pores of the axial prongs of the medial and lateral hooks are located well anterior to the middle of the prong in M. sumansinghai n. sp., they are well posterior to the middle of the prongs in M. shawae n. sp. In addition, the base of the lateral hook is longer relative to that of the medial hook in the latter species than it is in the former species. The proglottid anatomy of valid species of Megalonchos is described for the first time, and the lack of post-vaginal testes is confirmed for the genus. In addition, members of this genus appear to be characterised by a sacciform uterus that extends only to the level of the cirrus-sac and an ovary that is H-shaped in frontal view and bilobed in cross-section. Species of Megalonchos have now been reported from two of the eight known species of hemigaleid sharks. PMID- 17464484 TI - Nilonema gymnarchi Khalil, 1960 and N. senticosum (Baylis, 1922) (Nematoda: Dracunculoidea): Gondwana relicts? AB - Data-base searches of records of the helminth parasites of South American and African freshwater fishes revealed only two pairs of species from genera endemic to the two regions. One pair, species of the primitive amphilinid cestode genus Nesolecithus Donges & Harder, 1966, has already been designated as likely Gondwana relicts. The second pair are the philometrid nematodes Nilonema gymnarchi Khalil, 1960 from Gymnarchus niloticus Cuvier (Gymnarchidae) in Africa and N. senticosum (Baylis, 1922) from the South American fish Arapaima gigas (Cuvier) (Arapaimidae). Both species are partly redescribed on the basis of light and scanning electron microscopical observations of the type-specimens, and their relationships are discussed. In view of the fact that both hosts are basal teleosts of the order Osteoglossiformes and they are the same two fish which harbour the amphilinid cestodes already indicated as Gondwana relics, the nematode pair is also deemed to be relictual. However, these species are suggested as being of limited potential value for calibrating a molecular clock. PMID- 17464485 TI - Validation and redescription of Amblyomma romitii Tonelli-Rondelli, 1939 (Acari: Ixodidae). AB - The current synonymy of Amblyomma romitii Tonelli-Rondelli, 1939, a Neotropical species, with the Oriental species A. extraoculatum Neumann, 1899 is discussed. The type-specimens of both species were examined and their morphological differences determined. Considering their morphology and their different continental origins, we are reasonably certain that both taxa are valid. The type specimen of A. extraoculatum is a female labelled as being from Singapore. This is the only locality data on this tick species, which does not imply that it is restricted to Singapore. The geographical distribution of A. romitii includes Brazil (state of Para), French Guiana, British Guiana (now Guyana), Surinam and Venezuela. A redescription of the male and female types of A. romitii, based on light microscopy, is provided. In addition, the relationship of this species with other Neotropical and Asian species of Amblyomma Koch, 1844 is discussed. PMID- 17464486 TI - [Intoxication with a tricyclic antidepressant]. AB - A 48-year-old woman with a major depression and treatment with doxepin was found comatose in her flat. Her son last saw her 48 h prior to being found. On arrival of the emergency physician, she presented a generalized seizure. The patient underwent endotracheal intubation and mechanical ventilation due to respiratory insufficiency and severe cyanosis. Empty packages of tablets (doxepin ca. 4000 mg and zolpidem 100 mg) were found in the flat. On hospital admission the doxepin blood concentration was 1.2 microg/ml. No life-threatening arrhythmia occurred at any time. On the advice of the poison information center, hemoperfusion was performed for extracorporeal elimination. Within several hours the doxepin blood concentration could be lowered to 0.8 microg/ml and although still above the therapeutic range the patient was extubated. However, the patient developed a generalized seizure which required re-intubation. As a consequence of the high distribution volume and re-distribution phenomena, the doxepin blood concentration had increased again to 1.2 microg/ml. Approximately 72 h later she was extubated again while the doxepin blood concentration was 0.9 microg/ml and 3 days later, the doxepin blood concentration was lowered to 0.3 microg/ml and the patient was transferred to the psychiatric ward the following day. This case report questions the efficacy of hemoperfusion during acute doxepin intoxication in the given constellation of a non-life-threatening arrhythmia. PMID- 17464487 TI - [Statistical process control as a monitoring tool for the evaluation of reorganisation measures. Investigation in an intensive care unit]. AB - INTRODUCTION: The German health care system is currently in a constant state of flux owing to enhanced competition and to the increasing focus on economic aspects. Medical services, especially treatment processes, are being reorganised in an attempt to adapt them to the new economic challenges. Ideally, radical reorganisation and streamlining of medical therapy processes should be accompanied by controlling and quality management systems. The purpose of this is to monitor the intensity of any economic and any patient-related (side)-effects. Business management techniques are needed that allow online and long-term performance reviews of reorganisation measures once initiated. METHODS: In industry, the method applied for this purpose is statistical process control (SPC). The present study demonstrates for the first time that use of this monitoring tool can be extended to the medical sector. In an intensive care unit (ICU) the following process parameters were monitored: duration of sedation, time to persisting spontaneous breathing, length of stay in ICU, length of stay in hospital, patient mortality in ICU and in the next 30 days after admission to the ICU. Group 1 was made up of 87 patients examined before and group 2, 93 patients after process optimisation. The main feature of the reorganisation was application of a new analgo-sedation technique and of the weaning concept. RESULTS: In group 2 duration of sedation, time to spontaneous breathing and length of stay on the ICU were significantly shorter than in group 1. The length of stay in hospital, patient mortality in the ICU and 30 days after the initiation of intensive care did not differ significantly between the two groups. CONCLUSION: Economic and patient-related key figures can be evaluated with SPC. It allows online assessment both before and during process optimisation, and especially in the long term afterprocess optimisation. PMID- 17464488 TI - [Lactate and redox status in malignant tumors]. AB - This article summarizes data from experimental and clinical oncology which are indicative of a pivotal role of tumor carbohydrate metabolism in malignant behavior and outcome of treatment. In primary tumors, such as cervix carcinomas, head and neck squamous cell carcinomas or rectum adenocarcinomas, elevated lactate levels as a mirror of a high glycolytic activity, are correlated even at the initial diagnosis with a high level of malignancy as indicated by increased formation of metastases or an elevated radiotherapy resistance. The relationship between therapeutic resistance and glycolysis may at least partially be due to the radical scavenging potential of glycolytic intermediates, mainly pyruvate and lactate and to the link between these metabolites and the cellular redox status. On the basis of these data and other considerations, a novel technique has been developed for imaging the lactate/pyruvate ratio in tumor biopsies using quantitative bioluminescence. More research effort should, therefore, be focussed on the redox status of tumors in oncological studies in the future. PMID- 17464489 TI - [Erythema annulare centrifugum-like psoriasis cum pustulatione]. AB - Erythema annulare centrifugum-type psoriasis with pustules represents a subtype of psoriasis pustulosa generalisata von Zumbusch. It presents with a typical morphology characterized by a lack of classical erythematosquamous skin lesions during its acute eruption phase. Diagnosis is usually established on the basis of clinical presentation and dermatopathology, which often shows a substrate typical for psoriasis, sometimes with spongiform pustules. PMID- 17464490 TI - [Surgical correction of scleroderma en coup de sabre]. AB - A 15-year-old patient developed scleroderma en coup de sabre on right temple at 5 years of age. Multiple treatments (3 cycles of intravenous penicillin, topical glucocorticosteroids, topical calcipotriol, and cream PUVA phototherapy combined with topical calcipotriol) produced no improvement. The patient suffered greatly from the psychosocial stigmatization, so that the entire lesion was resected at 14 years of age. One year after the operation a thin non-sclerotic scar was present; tiny lateral areas of sclerosis not included in the operative field were unchanged. The operation greatly improved the patient's daily life. The surgical therapy of scleroderma en coup de sabre offers an interesting therapeutic alternative. PMID- 17464491 TI - [Liposarcoma: a rare differential diagnosis of cervical space occupation]. AB - With less than 100 cases described in the literature, the incidence of liposarcoma in the head and neck is very low. We report on a 54-year-old patient with a liposarcoma of the right cervical soft tissue. After complete tumor extirpation the patient underwent radiotherapy. Diagnostics, histopathological classification, therapy and prognosis of liposarcoma are discussed in this case report and review of the literature. PMID- 17464492 TI - [ENT and head and neck surgery in the German DRG system 2007]. AB - BACKGROUND: The German DRG system has been further developed into version 2007. For ENT and head and neck surgery, significant changes in the coding of diagnoses and medical operations as well as in the the DRG structure have been made. RESULTS: New ICD codes for sleep apnoea and acquired tracheal stenosis have been implemented. Surgery on the acoustic meatus, removal of auricle hyaline cartilage for transplantation (e. g. rhinosurgery) and tonsillotomy have been coded in the 2007 version. In addition, the DRG structure has been improved. Case allocation of more than one significant operation has been established. CONCLUSION: The G DRG system has gained in complexity. High demands are made on the coding of complex cases, whereas standard cases require mostly only one specific diagnosis and one specific OPS code. The quality of case allocation for ENT patients within the G-DRG system has been improved. Nevertheless, further adjustments of the G DRG system are necessary. PMID- 17464493 TI - [Cutaneous basosquamous cell carcinoma]. AB - BACKGROUND: Basosquamous carcinoma (BSC) is a rare malignancy with specific histopathological features of both basal cell (BCC) and squamous cell carcinoma (SCC). Therefore, the histological diagnosis is challenging. Due to its low incidence there is no consensus on the surgical management of BSC. PATIENTS AND METHODS: We describe the (immunohistological) diagnostic and therapeutic aspects of nine cases (8 male symbol : 1 female symbol, on average 68.6 years of age (range: 47-81)) of BSC. Of these, seven were located on the pinna, one on the forehead and zygomatic region and one in the retroauricular region. RESULTS: Immunochemical staining for epithelial membrane antigen was negative (apart from the typical areas of epithelial pearl formation) and BerEP4 was positive in all cases. Therapy consisted of partial removal of the pinna in four, and total removal in three cases. One patient was treated by partial removal of the auricle with superficial parotidectomy and ipsilateral neck dissection. In the case of the carcinoma on the forehead, a local excision was performed. The median follow up was 45 months. One patient had a local recurrence. CONCLUSIONS: The histological diagnosis of BSC is confirmed by the use of EMA and BerEP4 immunohistological staining. Clinically, BSC is a rare, aggressive skin tumor. Despite the histological similarity to basal cell carcinoma, BSC has an imminent risk of metastasizing. Hence, therapy should be similar to that for SCC, taking into consideration the age and general state of the usually elderly patient. PMID- 17464494 TI - [Anterior spondylosis of the cervical spine causing dyspnea and sleep disturbance]. AB - BACKGROUND: Cervical osteochondrosis is a rare differential diagnosis leading to dysphagia, inspiratory stridor and obstructive sleep apnea syndrome (OSAS). PATIENTS AND METHODS: We report six cases of patients with episodes of neck pain (n=6), pain reflected to the arm (n=1), sleep apnea (n=5), inspiratory stridor (n=3) and/or unclear dysphagia (n=6), who presented between 2000 and 2003 at the Romerwallklinik Mainz and the university hospitals of Mainz and Cologne. None of these patients had symptoms of spinal or radicular compression. All underwent otorhinolaryngological and radiological examination followed by excision of anterior spondylophytes and intervertebral fusion. One patient required immediate tracheotomy due to perforation of the pharyngeal wall associated with severe supraglottic swelling. RESULTS: All patients were free of inspiratory stridor postoperatively. Symptoms of dysphagia disappeared in four patients and were reduced in two. Three of five patients were free of apnea. OSAS had improved in two. Neck pain was eliminated in four cases and markedly improved in two cases. CONCLUSIONS: In case of symptoms of pharyngeal compression and OSAS, a vertebragenic cause should be considered. PMID- 17464495 TI - [Precision of guidewire placement--can it be improved by applying the new, isocentric aiming principle?]. AB - BACKGROUND: Exact placement of a guidewire is difficult for the less experienced surgeon as this complex 3D task usually is controlled by means of 2D fluoroscopic projections. The new isocentric aiming principle presented here splits up the 3D task into two planar, 2D steps. Movements of the guidewire to achieve correct placement are limited to one plane per step and can therefore be exactly controlled by fluoroscopy. The fluoroscopic projection needs to be changed only once in between the two steps. METHODS: The isocentric aiming principle became applicable to the proximal femur region by means of a mechanical aiming device. We have done an experimental study in order to compare the new isocentric aiming principle to the freehand aiming technique which is routinely applied. We documented the precision of guidewire placement achieved (angular deviation of the guidewire in two projections, linear deviation of the actual from the intended entry point), number of fluoroscopic controls, and procedure time when guidewire placement is done by an experienced and by an inexperienced surgeon. RESULTS: When applying the isocentric aiming principle the inexperienced surgeon succeeded in fixing the entry angle of the guidewire more precisely both in the AP [1.3 degrees (0.0-2.0 degrees ) versus 2.3 degrees (0.0-9.0 degrees ), p=0.034] as well as in the axial view [1.0 degrees (0.0-2.5 degrees ) versus 6.5 degrees (0.0-12.0 degrees ), p=0.036]. Linear displacement was not significantly different between the two methods: 4.4 (0.7-9.6) mm deviation with the isocentric aiming principle versus 3.9 (1.6-5.7) mm, p=0.406, when the freehand technique is applied. When applying the isocentric aiming principle for guidewire placement the experienced surgeon achieved less precise angulation in the AP view [2.5 degrees (0.0-4.0 degrees ) versus 1.8 degrees (0.0-3.5 degrees ), p=0.061], improved precision in the axial view [2.0 degrees (1.0-3.0 degrees ) versus 3.0 degrees (0.0-5.0 degrees ), p=0.074], and a slightly worsened linear displacement [2.5 (1.0-4.2) mm versus 2.0 (1.0-2.6) mm, p=0.131]. Both surgeons needed less fluoroscopic controls when using the isocentric aiming principle instead of the freehand aiming method: inexperienced surgeon: 8.0 controls (7.0-16.0) instead of 13.0 controls (7.0-16.0), p=0.043; experienced surgeon: 14.5 controls (8.0-26.0) instead of 16.5 controls (12.0-33.0), p=0.282. However due to the additional time needed to fix and align the aiming device to the bone both surgeons required increased procedure time when using the isocentric aiming principle: 4.3 (3.0 6.9) min instead of 2.6 (2.2-4.0) min, p=0.005, for the inexperienced surgeon and 3.3 (2.3-4.3) min instead of 1.9 (1.4-2.8) min, p=0.001, for the experienced surgeon. CONCLUSIONS: Based on the experimental results we would suggest clinical application of the isocentric aiming principle especially for the less experienced surgeon. Increased precision would outweigh the drawback of a slightly prolonged procedure time. X-ray exposure may also be reduced when using the isocentric aiming principle for guidewire placement. However our results have to be verified by a clinical study beforehand. The isocentric aiming principle can also be applied in other situations that allow for two orthogonal projections for guidewire placement. PMID- 17464496 TI - [Infanticide as a consequence of postpartum bonding disorder]. AB - Parental killing of their own children in the 1st year constitutes a rare phenomenon. Studies on infanticide show that mothers who kill their children are frequently psychiatrically disturbed. Depression is the most common postpartum disorder and may represent a vital danger for both mother and child. The association between depression and infanticide can usually be explained by maternal psychopathologic symptoms. The importance of a mother's bonding to her infant has not often been examined in respect to infanticide. The present case report underlines the importance of postpartum bonding disorder and its relation to a higher risk of infanticide. PMID- 17464497 TI - [Primary prevention of bladder cancer. What's new?]. AB - Bladder cancer (Bc) with an incidence of 8.6% ranks fourth in males and with an incidence of 3.5% ranks eighth in females in Germany. The study of Cole demonstrating that coffee drinking poses a risk for developing Bc raised interest in primary prevention. In the meantime, however, 42 case-control studies could disprove this finding. Cigarette smoking raises the Bc risk threefold. Responsible are among others arylamines which are activated in the liver, but also detoxified. A genetically caused lack of transferases is responsible for every third Bc. Another risk factor is contributed by permanent hair dyes. Similarly to smoking, an arylamine is taken up by the body and has to be detoxified by transferases in the liver and skin. Furthermore, a chronic urinary tract infection may be related to Bc. Thus, the best prevention is the reduction of risk factors.A high fluid intake--irrespective of its kind--reduces the Bc risk by approximately 50%. Particularly smokers should realize that they can lower their risk of developing Bc by almost 70% with a high fluid intake. A connection between alcohol consumption and Bc development has never been shown. Vitamins A, B, and C were extensively investigated in epidemiological studies. Usefulness for primary prevention of Bc was not convincingly demonstrated. Interestingly, folic acid-containing food may reduce the Bc risk for smokers. Fruits and vegetables possess only a weak preventive efficacy. However, copious fruit consumption may reduce the risk for smokers by almost 50%. The trace element selenium does not possess a proven protection; however, it has been shown that persons with a high selenium plasma level have a lower incidence of Bc. Among probiotics yogurt containing Lactobacillus casei is particularly useful in smokers according to one study. Finally, the preventive action of NSAIDs is controversial. Surprisingly, users of analgetics have a low incidence of Bc. PMID- 17464498 TI - A German genome-wide linkage scan for type 2 diabetes supports the existence of a metabolic syndrome locus on chromosome 1p36.13 and a type 2 diabetes locus on chromosome 16p12.2. AB - AIMS/HYPOTHESIS: The aim was to identify type 2 diabetes susceptibility regions in 250 German families. SUBJECTS AND METHODS: We conducted a genome-wide linkage scan using 439 short tandem repeat polymorphisms at an average resolution of 7.76 +/- 3.80 cM (Marshfield). In an affected-only-design (affected sib pairs), we performed nonparametric multipoint linkage analyses. Conditional analyses were applied where linkage signals were found in the baseline analyses. RESULTS: We identified two loci with nominal evidence for linkage on chromosomes 1p36.13 and 16p12.2 (D1S3669, 37.05 cM, logarithmic odds ratio [LOD] = 1.49, p = 0.004; D16S403, 43.89 cM, LOD = 1.85, p = 0.002). D16S403 crossed the empirically obtained threshold of genome-wide suggestive significance of LOD = 1.51. Positive findings in those regions have been reported by the following other linkage studies on: (1) symptomatic/clinical gall bladder disease with type 2 diabetes in Mexican Americans from the San Antonio Family Diabetes/Gallbladder Study (LOD = 3.7, D1S1597-D1S407, 29.93-33.75 cM); (2) body size-adiposity in another Mexican American population (D1S1597, LOD = 2.53, 29.93 cM); (3) lipid abnormalities (LOD = 3.1, D1S2826-D1S513, 41.92-60.01 cM); and (4) hypertension in Australian sib pairs (LOD = 3.1, D1S2834-D1S2728, 31.02-33.75 cM); as well as (5) a meta analysis of four European type 2 diabetes-related genome scans (LOD = 1.09, D16S412, 42.81 cM). In linkage analyses conditional on evidence for linkage at D16S403 we identified a LOD increase (Delta LOD) of 1.55 (p = 0.0075) at D17S2180. Similar conditioning on D17S2180 revealed evidence for interaction with D1S3669 (Delta LOD = 1.67, p = 0.0055), D16S403 (Delta LOD = 1.48, p = 0.0091) and another locus on chromosome 1 where several genome scans have reported evidence for linkage ( approximately 200 cM, Delta LOD = 1.60, p = 0.0066). CONCLUSIONS/INTERPRETATION: Our results and the findings of other studies are consistent with the presence of a locus for a complex metabolic syndrome on chromosome 1p36.13. PMID- 17464499 TI - Fluctuation in recoverable nickel and zinc resistant copiotrophic bacteria explained by the varying zinc ion content of Torsa River in different months. AB - Heavy metal content analysis of River Torsa in India did not indicate any alarming level of toxicity for human consumption but revealed variation at the ppb level in different months. The variation in recoverable nickel and zinc resistant copiotrophic (or eutrophic) bacterial counts was explained by the variation of the zinc content (34.0-691.3 ppb) of the river water in different sampling months. Growth studies conducted with some purified nickel and/or zinc resistant strains revealed that pre-exposure of the cells to ppb levels of Zn(2+), comparable to the indigenous zinc ion concentration of the river, could induce the nickel or zinc resistance. A minimum concentration of 5-10 microM Zn(2+ )(325-650 ppb) was found effective in inducing the Nickel resistance of the isolates. Zinc resistance of the isolates was tested by pre-exposing the cells to 4 microM Zn(2+ )(260 ppb). The lag phase was reduced by 6-8 h in all the cases. Biochemical characteristics and phylogenetic analysis based on 16S rDNA sequence indicated that some of the Torsa River isolates, having inducible nickel and zinc resistance, are members of the genus Pseudomonas, Acinetobacter, Bacillus, Enterobacter, Serratia and Moraxella. PMID- 17464500 TI - Comparative study of activities in reactive oxygen species production/defense system in mitochondria of rat brain and liver, and their susceptibility to methylmercury toxicity. AB - The involvement of oxidative stress has been suggested as a mechanism for neurotoxicity caused by methylmercury (MeHg), but the mechanism for MeHg selective toxicity in the central nervous system is still unclear. In this research, to clarify the mechanism of selective neurotoxicity caused by MeHg, the oxygen consumption levels, the reactive oxygen species (ROS) production rates and several antioxidant levels in mitochondria were compared among the cerebrum, cerebellum and liver of male Wistar rats. In addition, the alterations of these indexes were examined in MeHg-intoxicated rats (oral administration of 10 mg/kg day, for 5 days). Although the cerebrum and cerebellum in intact rats showed higher mitochondrial oxygen consumption levels and ROS production rates than the liver, glutathione peroxidase (GPX) and superoxide dismutase (SOD) activities were much lower in the cerebrum and cerebellum than in the liver. Especially, the cerebellum showed the highest oxygen consumption and ROS production rate and the lowest mitochondrial glutathione (GSH) levels among the tissues examined. In the MeHg-treated rats, decrease in the oxygen consumption and increase in the ROS generation were found only in the cerebellum mitochondria, despite a lower Hg accumulation in the mitochondrial fraction compared to the liver. Since MeHg treatment produced an enhancement of ROS generation in cerebellum mitochondria supplemented with succinate substrates, MeHg-induced oxidative stress might affect the complex II-III mediated pathway in the electron transfer chain in the cerebellum mitochondria. Our study suggested that inborn factors, high production system activity and low defense system activity of ROS in the brain, would relate to the high susceptibility of the central nervous system to MeHg toxicity. PMID- 17464502 TI - Choroid plexus papilloma of the third ventricle. PMID- 17464501 TI - Habit formation and the loss of control of an internal clock: inverse relationship between the level of baseline training and the clock-speed enhancing effects of methamphetamine. AB - INTRODUCTION: Drugs that modulate the effective level of dopamine (DA) in cortico striatal circuits have been shown to alter the perception of time in the seconds to-minutes range. How this relationship changes as a function of repeated experience with the reinforcement contingencies and the gradual adaptation of the underlying neural circuits remains unclear. MATERIALS AND METHODS: The present study examined the clock-speed enhancing effects of methamphetamine (MAP 0.5 or 1.0 mg/kg, ip) in groups of rats that received different levels of baseline training (20, 60, or 120 sessions) on a 50-s peak-interval (PI) procedure before initial drug administration. RESULTS: A curvilinear relationship was observed such that rats that received either minimal or intermediate levels of training (or=120 sessions) did not show this "classic" DA agonist curve-shift effect, but instead displayed a dose-dependent disruption of temporal control after MAP administration. A transition from DA-sensitive to DA-insensitive mechanisms is proposed to account for the loss of control of clock speed, as timing behaviors associated with the PI procedure gradually become learned habits through the strengthening of DA-glutamate connections. PMID- 17464503 TI - Osteosarcomatosis. PMID- 17464504 TI - High-throughput killer cell immunoglobulin-like receptor genotyping by MALDI-TOF mass spectrometry with discovery of novel alleles. AB - The killer cell immunoglobulin-like receptors (KIR) interact with major histocompatibility complex (MHC) class I ligands to regulate the functions of natural killer cells and T cells. Like human leukocyte antigens class I, human KIR are highly variable and correlated with infection, autoimmunity, pregnancy syndromes, and transplantation outcome. Limiting the scope of KIR analysis is the low resolution, sensitivity, and speed of the established methods of KIR typing. In this study, we describe a first-generation single nucleotide polymorphism (SNP)-based method for typing the 17 human KIR genes and pseudogenes that uses analysis by matrix-assisted laser desorption/ionization time-of-flight (MALDI TOF) mass spectrometry. It is a high-throughput method that requires minute amounts of genomic DNA for discrimination of KIR genes with some allelic resolution. A study of 233 individuals shows that the results obtained by the SNP based KIR/MALDI-TOF method are consistent with those obtained with the established sequence-specific oligonucleotide probe or sequence-specific polymerase chain reaction methods. The added sensitivity of the KIR/MALDI-TOF method allowed putative novel alleles of the KIR2DL1, KIR3DL1, KIR2DS5, and KIR2DL5 genes to be identified. Sequencing the KIR2DL5 variant proved it was a newly discovered allele, one that appears associated with Hispanic and Native American populations. This KIR/MALDI-TOF method of KIR typing should facilitate population and disease-association studies that improve knowledge of the immunological functions of KIR-MHC class I interactions. PMID- 17464505 TI - Radionuclide reporter gene imaging for cardiac gene therapy. AB - INTRODUCTION: In the field of cardiac gene therapy, angiogenic gene therapy has been most extensively investigated. The first clinical trial of cardiac angiogenic gene therapy was reported in 1998, and at the peak, more than 20 clinical trial protocols were under evaluation. However, most trials have ceased owing to the lack of decisive proof of therapeutic effects and the potential risks of viral vectors. In order to further advance cardiac angiogenic gene therapy, remaining open issues need to be resolved: there needs to be improvement of gene transfer methods, regulation of gene expression, development of much safer vectors and optimisation of therapeutic genes. For these purposes, imaging of gene expression in living organisms is of great importance. In radionuclide reporter gene imaging, "reporter genes" transferred into cell nuclei encode for a protein that retains a complementary "reporter probe" of a positron or single photon emitter; thus expression of the reporter genes can be imaged with positron emission tomography or single-photon emission computed tomography. Accordingly, in the setting of gene therapy, the location, magnitude and duration of the therapeutic gene co-expression with the reporter genes can be monitored non invasively. In the near future, gene therapy may evolve into combination therapy with stem/progenitor cell transplantation, so-called cell-based gene therapy or gene-modified cell therapy. CONCLUSION: Radionuclide reporter gene imaging is now expected to contribute in providing evidence on the usefulness of this novel therapeutic approach, as well as in investigating the molecular mechanisms underlying neovascularisation and safety issues relevant to further progress in conventional gene therapy. PMID- 17464506 TI - Integration of genomics, proteomics, and imaging for cardiac stem cell therapy. AB - Cardiac stem cell therapy is beginning to mature as a valid treatment for heart disease. As more clinical trials utilizing stem cells emerge, it is imperative to establish the mechanisms by which stem cells confer benefit in cardiac diseases. In this paper, we review three methods - molecular cellular imaging, gene expression profiling, and proteomic analysis - that can be integrated to provide further insights into the role of this emerging therapy. PMID- 17464507 TI - Identification of a new hTERT-derived HLA-A*0201 restricted, naturally processed CTL epitope. AB - By the use of a neural network capable of performing quantitative predictions of peptides binding to HLA-A*0201 molecules, we identified a number of nonamer peptides derived from the catalytic subunit of telomerase, human telomerase reverse transcriptase (hTERT). Five nonimmunogenic peptides with measured binding affinities for HLA-A*0201 ranging from 155 to 1,298 nM were modified at the P1, P2 and P9 positions, respectively, to achieve stronger HLA-A*0201 binding. One peptide, mp30-38 (mp30), with an L to V substitution at position 9 was subsequently found to be immunogenic in mp30 immunized HLA-A*0201/H2K(b) or HHD transgenic mice. The T cell reactivity obtained was directed against both the mp30 and against the unmodified p30. Anti-mp30 specific T cells generated in HLA A*0201 transgenic mice were dependent on TCR-CD8/MHC-I alpha3 binding and therefore not capable of recognizing mp30-pulsed human HLA-A*0201(+) cells or murine HLA-A*0201 transfectants. In order to show reactivity against naturally processed peptide in human tumor cells, an hTERT positive HLA-A*0201 negative colon carcinoma cell line (CCL220) was transfected with an HLA-A*0201/H2K(b) cDNA construct and used as target in ELISPOT and cytotoxicity assays. The data show that T cells from mp30 immunized HHD transgenic mice react specifically against the CCL220 transfectant indicating that p30 is naturally processed. In conclusion, we have identified a new CTL HLA-A*0201 restricted hTERT epitope, which is now, included in an ongoing phase 2 vaccine trial of patients with disseminated cancer. PMID- 17464508 TI - Proximal bone remodelling differed between two types of titanium long femoral components after cementless revision arthroplasty. AB - In revision surgery with proximal femoral bone loss, progressive bone atrophy due to stress shielding remains a concern. We compared 2-year radiological results between two types of cementless long titanium stems with different configurations and surface coatings. Of 17 hips implanted with a wholly hydroxyapatite-coated stem, 12 (71%) exhibited stress shielding of the second degree or higher according to Engh's criteria, and the mean relative bone mass index decreased from 22.1% pre-operatively to 14.6% at 2 years post-operatively. In 23 hips implanted with a sand-blasted, conically shaped stem, no hip showed stress shielding of the second degree or higher. The mean relative bone mass index increased from 21.6% to 31.4%. These results indicate that the configuration and surface coating of the stem have a significant influence on proximal bone remodelling after revision surgery. PMID- 17464509 TI - Vascular activity of two silicon compounds, ALIS 409 and ALIS 421, novel multidrug-resistance reverting agents in cancer cells. AB - PURPOSE: The aim of this study was to investigate the effects of two novel multidrug-resistance reverting agents, ALIS 409 [1,3-dimethyl-1,3-p-fluorophenyl 1,3(3-morfolinopropyl)-1,3-disiloxan dihydrochloride] and ALIS 421 [1,3-dimethyl 1,3-(4-fluorophenyl)-1,3[3(4-buthyl)-(1-piperazinyl)-propyl]-1,3-disiloxan tetrahydrochloride], on vascular functions in vitro. EXPERIMENTAL DESIGN: A comparison of their mechanical and electrophysiological actions in rat aorta rings and single rat tail artery myocytes, respectively, was performed. RESULTS: In endothelium-denuded rat aorta rings, ALIS 409 and ALIS 421 antagonized 60 mM K(+)-induced contraction in a concentration-dependent manner with IC(50) values of 52.2 and 15.5 microM, respectively. ALIS 409 and ALIS 421 inhibited L-type Ca(2+) current recorded in artery myocytes in a concentration-dependent manner with IC(50) values of 6.4 and 5.6 microM, respectively. In rat aorta, ALIS 409 and ALIS 421 antagonized the sustained tonic contraction induced by phenylephrine with IC(50) values of 58.0 and 13.7 microM (endothelium-denuded rings) and of 73.9 and 31.9 microM (endothelium-intact rings), respectively. In endothelium denuded rings, ryanodine reduced significantly the response to phenylephrine in the absence of extracellular Ca(2+) whereas nifedipine, ALIS 409 or ALIS 421 did not affect it. Phenylephrine-stimulated influx of extracellular Ca(2+) was markedly reduced when tissues were pretreated with ALIS 409, ALIS 421 or nifedipine, and stimulated when they were pretreated with ryanodine. Application of ALIS 409 (up to 100 microM) to intact rat aorta rings failed to induce mechanical responses. CONCLUSIONS: Our results provide functional evidence that the myorelaxing effect elicited either by ALIS 409 or by ALIS 421 involved mainly the direct blockade of extracellular Ca(2+) influx. This effect, however, took place at concentrations much higher than those effective as modifiers of multidrug resistance in cancer cells. PMID- 17464510 TI - Complex conotruncal cardiac anomalies consecutively in three siblings from a consanguineous family possibly associated with maternal hyperhomocysteinemia. AB - INTRODUCTION: Conotruncal defects represent an anatomically heterogeneous group of cardiac malformations affecting the outflow tract of the ventricles and the arterial pole of the heart. The exact etiology of congenital heart diseases is unknown. CASE REPORT: A 31-year-old woman who had three offspring with complex conotruncal cardiac anomalies after consanguineous marriage was reported. The first child is still alive. However, the second affected child died at the age of 3 years. Fluorescence in situ hybridization studies of the siblings excluded CATCH(22) chromosomal abnormality. The maternal laboratory work-up was unremarkable except for low serum folic acid and cobalamin levels and high homocysteine levels. The woman received high dose pyridoxine, cobalamin and folate treatment preconceptionally, and she became pregnant. She delivered a healthy male infant without any abnormalities. Six months later, she became pregnant again without any preconceptional medications. When she was examined during the fourth pregnancy, unfortunately the fetus was found to have a restrictive ventricular septal defect, a right ventricle with two outflows and a right aortic arch. Her last pregnancy was terminated at 22 weeks. The 22-week-old female fetus was examined postmortem and the diagnosis of congenital heart disease was confirmed. CONCLUSION: Cobalamin and folate administration may help to reduce the development of cardiac malformations. PMID- 17464511 TI - A randomized, double-blind, parallel-group study to compare the efficacy and safety of a once-daily loratadine-pseudoephedrine combination with that of a twice-daily loratadine-pseudoephedrine combination in the treatment of allergic rhinitis. AB - The aim of this study is to compare the efficacy and safety of a once-daily loratadine-pseudoephedrine sustained release tablet with that of a twice-daily loratadine-pseudoephedrine in the treatment of patients with allergic rhinitis. Forty-eight subjects were randomized into either the treatment or control group. The efficacy endpoint was assessed by total symptom scores (TSS). And the results were analyzed by non-inferiority testing and t-tests. Non-inferiority testing of the once daily loratadine 10 mg-pseudoephedrine 240 mg sustained release tablets to the twice-daily loratadine 5 mg-pseudoephedrine 120 mg combination tablets was not supported by statistical significance. However, both the treatment and control groups showed a significant reduction from the baseline in TSS (P < 0.05), and the difference between groups did not reach statistical significance (P > 0.05). In conclusion, once-daily and twice-daily preparations of loratadine pseudoephedrine were comparable in efficacy and safety in the treatment of allergic rhinitis. PMID- 17464512 TI - Acute interstitial nephritis of HIV-positive patients under atazanavir and tenofovir therapy in a retrospective analysis of kidney biopsies. AB - We performed a retrospective analysis on kidney biopsies of 30 human immunodeficiency virus (HIV)-positive patients. Twenty-two of them received highly active antiretroviral therapy (HAART). Tenofovir containing HAART together with atazanavir, a new protease inhibitor, was administered to three patients. All of them developed acute renal failure. The kidney biopsies of these patients showed an acute interstitial nephritis or a chronic interstitial nephritis with an acute component. Withdrawal of atazanavir and tenofovir resulted in recovery of renal function in all three patients. Acute interstitial nephritis was observed only in 1 of 19 patients without atazanavir or tenofovir treatment. We conclude that acute interstitial nephritis and consecutive acute renal failure is a relevant side effect of atazanavir and tenofovir therapy in HIV-positive patients. PMID- 17464513 TI - Immunohistochemical study on caveolin-1alpha in regenerating process of tubular cells in gentamicin-induced acute tubular injury in rats. AB - Caveolin-1, a principal component of caveolae, modulates growth signaling, endocytosis, and intracellular transport. We examined the expression of caveolin 1alpha and its relation to cell cycle and caveolin-interacting growth factor receptors in regenerating proximal tubules (PTs) after gentamicin-induced acute renal failure in rats. Caveolin-1alpha appeared in regenerating PTs as early as day 4 after last gentamicin, peaked at days 6 to 8, and showed cytoplasmic pattern after day 8. Immunoelectron microscopy revealed caveolin-1alpha-positive caveolae on the cell membrane and in cytoplasms in regenerating PTs at days 4 to 8 and caveolin-positivity confined to cytoplasms after day 10. The number of PT cells with proliferation markers peaked at day 6 and decreased afterwards as expression of cyclin-dependent kinase inhibitors increased. Platelet-derived growth factor receptor-beta (PDGFR-beta) and epidermal growth factor receptor (EGFR) were colocalized with caveolin-1alpha in proliferating PTs as early as day 4. Phosphorylated EGFR increased at day 8 and afterwards when caveolins dissociated from EGFR or decreased. In case of PDGFR-beta, phosphorylation seemed to be associated with the increase and association of caveolins to the receptors. Our results suggest that transient expression of caveolin-1alpha in early regenerating PTs might contribute to the regenerating process of PTs through modulating growth factor receptors. PMID- 17464514 TI - Hepatitis C virus infection acquired in childhood. AB - Hepatitis C virus (HCV) infection occurs less frequently in children than in adult patients, and the natural history, prognosis, and clinical significance of HCV infection in children are poorly defined. We report here a descriptive follow up of the clinical course, biochemical data, and viral markers observed in 37 children with anti-HCV. Ten patients included in the study tested persistently negative for serum HCV-RNA (group 1) and 27 patients tested persistently positive (group 2). In group 1, serum alanine aminotransferase (ALT) was normal in all patients, while two patients had non-organ-specific autoantibodies. In group 2, serum ALT was elevated in 13 of 27 patients, and five patients had non-organ specific autoantibodies. HCV genotype 1a and 1b were the most prevalent among HCV RNA-positive patients. Twenty liver biopsies were carried out on 17 patients in our series (mean evolution time, 11.2 years; range, 3-21 years). The liver specimens showed mild necroinflammatory changes in most patients, and fibrosis was absent or low grade. Two HCV-RNA-positive patients became persistently HCV RNA negative. Of the 26 children investigated, 7 (one in group 1, six in group 2) had a co-infection with hepatitis G virus. Conclusion Most children chronically infected with HCV were asymptomatic and presented only mild biochemical evidence of hepatic injury. Autoimmunity in the form of non-organ-specific autoantibodies was common. HCV in children induced mild changes in the liver with a low level of fibrosis and at a low rate of progression. PMID- 17464515 TI - Pathophysiology of hypercalciuria in children. AB - Urinary excretion of calcium is the result of a complex interplay between three organs-namely, the gastrointestinal tract, bone, and kidney-which is finely orchestrated by multiple hormones. Hypercalciuria is believed to be a polygenic trait and is influenced significantly by diet. This paper briefly reviews calcium handling by the renal tubule in normal and in hereditary disorders as it relates to the pathophysiology of hypercalciuria. The effects of dietary sodium, potassium, protein, calcium, and phosphate on calcium excretion, and the association of hypercalciuria with bone homeostasis is discussed, leading to recommendations on means to address excessive urinary calcium excretion. PMID- 17464516 TI - The quantitative measurements of the intervertebral angulation and translation during cervical flexion and extension. AB - The insufficient exploration of intervertebral translation during flexion and extension prevents the further understanding of the cervical biomechanics and treating the cervical related dysfunction. The objective of this study was to quantitatively measure the continuous intervertebral translation of healthy cervical spine during flexion and extension by videofluoroscopic technique. A total of 1,120 image sequences were analyzed for 56 healthy adult subjects by a precise image protocol during cervical flexion and extension. O: ur results showed there were no statistical angular differences among five spinal levels in either flexion or extension, except for the comparison between C2/3 (13.5 degrees) and C4/5 (22.6 degrees) angles. During cervical flexion, the smallest anterior translations were 0.7 mm at C2/3 level, followed by 0.9 mm at C6/7, 1.0 mm at C3/4, 1.1 mm at C5/6, and the largest 1.2 mm at C4/5 levels. The significantly greater translations were measured in the posterior direction at C3/4 (1.1 mm, P = 0.037), C4/5 (1.3 mm, P = 0.039), and C5/6 (1.2 mm, P = 0.005) levels than in the anterior one. The relatively fluctuant and small average posterior translation fashion at C6/7 level (0.4 mm) possibly originated from the variations in the direction of translation during cervical extension among subjects. Normalization with respect to the widths of individual vertebrae showed the total translation percentages relative to the adjacent vertebrae were 9.5, 13.7, 16.6, 15.0, and 8.6% for C2/3 to C6/7 levels, respectively, and appeared to be within the clinical-accepted ranges of translation in cervical spine. The intervertebral translations of cervical spine during flexion and extension movements were first described in quality and quantity based on the validated radiographic protocol. This analysis of the continuous intervertebral translations may be further employed to diagnose translation abnormalities like hypomobility or hypermobility and to monitor the treatment effect on cervical spines. PMID- 17464517 TI - Anterior lumbar instrumentation improves correction of severe lumbar Lenke C curves in double major idiopathic scoliosis. AB - Fifteen skeletally immature patients with double major adolescent idiopathic scoliosis with large lumbar curves and notable L4 and L5 coronal plane obliquity were retrospectively studied. Seven patients who underwent anterior release and fusion of the lumbar curve with segmental anterior instrumentation and subsequent posterior instrumentation ending at L3 were compared with eight patients treated with anterior release and fusion without anterior instrumentation followed by posterior instrumentation to L3 or L4. At 4.5 years follow-up (range 2.5-7 years), curve correction, coronal balance and fusion rate were not statistically different between the two groups; however, the group with anterior instrumentation had improved coronal plane, near normalangulation in the distal unfused segment compared with the group without anterior instrumentation. In cases involving severe lumbar curvatures in the context of double major scoliosis, when as a first stage anterior release is chosen, the addition of instrumentation appears to restore normal coronal alignment of the distal unfused lumbar segment, and may in certain cases save a level compared with traditional fusions to L4. PMID- 17464518 TI - Experimental evidence for route integration and strategic planning in wild capuchin monkeys. AB - Both in captivity and the wild, primates are found to travel mostly to the nearest available resource, but they may skip over the closest resource and travel to more distant resources, which are often found to be more productive. This study examines the tradeoff between distance and reward in the foraging choices of one group of wild capuchin monkeys (Cebus apella nigritus) using feeding platforms in large-scale foraging experiments conducted over four years. Three feeding sites were arrayed in an oblique triangle, such that once the monkey group had chosen one site to feed, they had a choice between two remaining sites, a close one with less food and the other up to 2.3 times as far away but with more food. Sites were provisioned once per day. The capuchins generally chose the closer feeding site, even when the more distant site offered up to 12 times as much food. The distances to, rewards of, or various profitability measures applied to each alternative site individually did not explain the group's choices in ways consistent with foraging theory or principles of operant psychology. The group's site choices were predicted only by comparing efficiency measures of entire foraging pathways: (1) direct travel to the more rewarding distant site, versus (2) the longer 'detour' through the closer site on the way to the more distant one. The group chose the detour more often when the reward was larger and the added detour distance shorter. They appeared to be more sensitive to the absolute increase in detour distance than to the relative increase compared to the straight route. The qualitative and quantitative results agree with a simple rule: do not use the detour unless the energy gain from extra food outweighs the energy cost of extra travel. These results suggest that members of this group integrate information on spatial location, reward, and perhaps potential food competition in their choice of multi-site foraging routes, with important implications for social foraging. PMID- 17464519 TI - Position paper on the use of mandibular advancement devices in adults with sleep related breathing disorders. A position paper of the German Society of Dental Sleep Medicine (Deutsche Gesellschaft Zahnaerztliche Schlafmedizin, DGZS). AB - Custom-made mandibular advancement devices are an effective treatment option for snoring, upper airway resistance syndrome, and obstructive sleep apnea (OSA). Evidence-based data indicates their efficacy, and international sleep societies recommend oral appliance (OA) therapy for patients with sleep-related breathing disorders. The following position paper by the German Society of Dental Sleep Medicine (DGZS) is to guide the interdisciplinary team (sleep physician and sleep disorder dentist) in detail when to prescribe oral appliances. This position paper supports the responsible use of OA as an effective treatment option for patients with sleep-related breathing disorders. The paper advises of proper indication regarding OSA severity, body mass index (BMI), and dentition. It emphasizes the interdisciplinary approach of oral appliance therapy and suggests treatment under the guidance of dentists trained in dental sleep medicine. PMID- 17464520 TI - Membrane fatty acids as pacemakers of animal metabolism. AB - The recent discovery that the fatty acid composition of tissue phospholipids varies in a systematic manner among species has lead to the proposal that membrane fatty acid composition is an important determinant of the metabolic rate characteristic for each species. Endotherms (mammals and birds) have a basal metabolic rate (BMR) that is several times that of ectotherms and have more polyunsaturated membranes. In both birds and mammals, as species size increases there is a decrease in mass-specific BMR and a decrease in membrane polyunsaturation. Membrane-associated processes are significant components of BMR and important membrane proteins operate at much faster rates in species with high BMR than in those with low BMR. A series of "species-crossover" experiments show that the rate of this molecular activity is largely due to the nature of the membrane bilayer surrounding these membrane proteins such that polyunsaturated membranes are associated with fast membrane-associated processes. It is suggested that this influence is due to the physical properties that such polyunsaturated membranes possess. This has been called the membrane pacemaker theory of metabolism and provides a framework to understand factors such as the influence of diet on metabolism. It is noted that in the rat membrane fatty acid composition is a regulated parameter being more influenced by the balance between n-3 and n-6 polyunsaturates in the diet than it is by general diet content of saturated, monounsaturated and total polyunsaturated fats. PMID- 17464521 TI - Authenticating production origin of gilthead sea bream (Sparus aurata) by chemical and isotopic fingerprinting. AB - Recent EU legislation (EC/2065/2001) requires that fish products, of wild and farmed origin, must provide consumer information that describes geographical origin and production method. The aim of the present study was to establish methods that could reliably differentiate between wild and farmed European gilthead sea bream (Sparus aurata). The methods that were chosen were based on chemical and stable isotopic analysis of the readily accessible lipid fraction. This study examined fatty acid profiles by capillary gas chromatography and the isotopic composition of fish oil (delta(13)C, delta(18)O), phospholipid choline nitrogen (delta(15)N) and compound specific analysis of fatty acids (delta(13)C) by isotope ratio mass spectroscopy as parameters that could reliably discriminate samples of wild and farmed sea bream. The sample set comprised of 15 farmed and 15 wild gilthead sea bream (Sparus aurata), obtained from Greece and Spain, respectively. Discrimination was achieved using fatty acid compositions, with linoleic acid (18:2n-6), arachidonic acid (20:4n-6), stearic acid (18:0), vaccenic acid (18:1n-7) and docosapentaenoic acid (22:5n-3) providing the highest contributions for discrimination. Principle components analysis of the data set highlighted good discrimination between wild and farmed fish. Factor 1 and 2 accounted for >70% of the variation in the data. The variables contributing to this discrimination were: the fatty acids 14:0, 16:0, 18:0, 18:1n-9, 18:1n-7, 22:1n-11, 18:2n-6 and 22:5n-3; delta(13)C of the fatty acids 16:0, 18:0, 16:1n-7, 18:1n-9, 20:5n-3 and 22:6n-3; Bulk oil fraction delta(13)C; glycerol/choline fraction bulk delta(13)C; delta(15)N; % N; % lipid. PMID- 17464523 TI - Differential changes in long-interval intracortical inhibition and silent period duration during fatiguing hand exercise. AB - During fatiguing exercise corticomotor excitability increases as force declines, which may serve to increase motor output to the exercising muscle, but paradoxically at the same time there is an increase in silent period (SP) duration which is thought to represent a build-up of intracortical inhibition. Paired-pulse TMS at long interstimulus intervals can also be used to derive an index of long-interval cortical inhibition (LICI), however this has not yet been investigated in fatigue. Our aim was to measure LICI during and after a fatiguing exercise and determine if the changes in the index of LICI parallel the changes in SP duration. To do this, we used single and paired-pulse TMS to measure motor evoked potential (MEP) amplitude, LICI and SP duration during, and for 10 min after, a 10-min intermittent maximal fatiguing exercise of the index finger, designed to fatigue the first dorsal interosseous (FDI) muscle (force after 10 min of exercise 64 +/- 7% of baseline, P < 0.05). Single-pulse MEP amplitude and SP duration were increased during fatiguing exercise (minute 10; 179 +/- 24% and 128 +/- 9% of baseline, respectively, P < 0.05), in contrast the measure of LICI was reduced compared to baseline (minute 10; 0.45 +/- 0.17 vs. baseline; 0.70 +/- 0.10, P < 0.05). These results suggest that SP duration and LICI may reflect processes occurring in different neuronal populations. The increased SP duration may correspond to processes of central fatigue in centres 'upstream' of primary motor cortex (M1), whereas the decrease in LICI, together with increased MEP amplitude, are consistent with an increase in M1 output during fatigue that may serve to compensate for reduced central drive. PMID- 17464524 TI - Role of PKC-delta as a signal mediator in epidermal barrier homeostasis. AB - The skin shows an important "epidermal permeability barrier homeostasis" in response to barrier disruption. Calcium ion (Ca(2+)), a major regulator in keratinocyte differentiation and proliferation, plays a crucial role in skin barrier homeostasis. Acute barrier disruption induces an immediate depletion of both extra- and intracellular calcium ions in the epidermis, especially in the upper granular layers, and results in the loss of normal epidermal calcium gradient. Currently, we hypothesize that the change in the intracellular calcium ion concentration triggers the barrier repair responses, such as lamellar body (LB) secretion and increased lipid synthesis in the epidermis. In this article, we suggest that PKC-delta is a signaling mediator for the changes in extracellular and intracellular calcium ion concentration. PMID- 17464525 TI - An approach for identifying postmenopausal women age 50-64 years at increased short-term risk for osteoporotic fracture. AB - Using data from NORA, we used 18 potential risk factors in a classification and regression tree analysis to build two algorithms. These algorithms correctly identified postmenopausal women between the ages of 50 and 64 years who were at the highest risk of osteoporotic fracture within 36 months. INTRODUCTION: The objective was to develop algorithms that best predict short-term fracture risk (3 years) in postmenopausal women 50-64 years old. METHODS: Data were from 91,652 women who responded to follow-up surveys as part of National Osteoporosis Risk Assessment (NORA) study. Peripheral bone mineral density (BMD) and risk factors obtained at baseline; incident osteoporotic fractures obtained from follow-up surveys. Eighteen risk factors were entered into a classification and regression tree analysis to build two algorithms, one with and one without BMD. RESULTS: Two thousand and seven (2.2%) women reported new osteoporotic fractures. Prior fracture, a peripheral BMD T-score or = 40% lower during the post-rehabilitation period than before. Physical effects of habitat rehabilitation were persistent through time, with pool habitat availability much higher in rehabilitated reaches than elsewhere. Fish community structure responded with major shifts in relative species abundance: as pool habitats increased after rehabilitation, small-bodied generalists and opportunists declined as certain piscivores and larger-bodied species such as centrarchids and catostomids increased. Reaches without habitat rehabilitation were significantly shallower, and fish populations there were similar to the rehabilitated reaches prior to treatment. These findings are applicable to incised, warmwater streams draining agricultural watersheds similar to those we studied. Rehabilitation of warmwater stream ecosystems is possible with current knowledge, but a major shift in stream corridor management strategies will be needed to reverse ongoing degradation trends. Apparently, conventional channel erosion controls without instream habitat measures are ineffective tools for ecosystem restoration in incised, warmwater streams of the Southeastern U.S., even if applied at the watershed scale and accompanied by significant reductions in suspended sediment concentration. PMID- 17464529 TI - Maintenance of forest biodiversity in a post-Soviet governance model: perceptions by local actors in Lithuania. AB - Successful biodiversity conservation does not depend on ecologic knowledge alone. Good conservation policies and policy implementation tools are equally important. Moreover, the knowledge, skills, and attitudes of local actors, directly in charge of operations in the field, are a key to successful policy implementation. The connections between policy objectives and their implementation as well as the involvement of local actors' efforts in implementing policy objectives largely depend on the governance model in use. This article assesses the knowledge of local actors in relation to the biodiversity conservation objectives and tools in Lithuanian forest management. As a main framework for this study, the needs assessment approach was applied. The study used both in-depth open-ended interviews and follow-up telephone interviews. Two state forest enterprises in Lithuania were selected as the study sites. The findings indicate that policy objectives in the field of forest biodiversity conservation and the related tools are well known but not well understood by those in charge of forest biodiversity policy implementation. To improve the situation, a transition toward adaptive learning and participatory governance as a means of facilitating conservation efforts is proposed. PMID- 17464532 TI - Surgery in intra-abdominal ruptured hydatid cyst. PMID- 17464533 TI - Can nipple-sparing mastectomy and immediate breast reconstruction with modified extended latissimus dorsi muscular flap improve the cosmetic and functional outcome among patients with breast carcinoma? AB - BACKGROUND: Nipple-areola complex (NAC) preservation is a new revolution in breast cancer surgery and breast reconstruction, if reliability and safety are considered. The latissimus dorsi muscular flap is a versatile flap that is gaining renewed popularity for immediate breast reconstruction with development of modifications. We are introducing nipple-sparing mastectomy (NSM) for Egyptian patients with breast carcinoma and reporting our results with a new modification of the extended latissimus dorsi muscular flap. METHODS: Between July 2005 and August 2006; forty-one patients with stage I to III breast carcinoma had NSM and immediate breast reconstruction. We performed a new modification to the extended latissimus dorsi muscular flap that allowed us to obtain enough autologous tissue to reconstruct the breast without implant or back incision. The postoperative aesthetic results with specific view of the preserved NAC were evaluated. RESULTS: We applied both an objective and subjective aesthetic result to our monitoring. Aesthetic grading results of breast reconstruction were as follows: excellent in 31, good in 6, fair in 2, poor in 2. Both reconstructed breast and donor site complications were minor. Patients are followed for a median follow-up of 7.9 months (range: 4-11 months). In this short period of follow-up, no local recurrence or distant failure has been observed. CONCLUSIONS: Nipple-sparing mastectomy with immediate breast reconstruction using modified extended latissimus dorsi muscular flap allows single-stage, totally autologous reconstruction with a satisfactory aesthetic result, low morbidity, and good quality of life. PMID- 17464536 TI - Re: Multiple Z-plasty in pilonidal sinus--a new technique under local anesthesia. PMID- 17464537 TI - Peritoneal perforation of liver hydatid cysts: clinical presentation, predisposing factors, and surgical outcome. AB - The aim of this study was to evaluate the clinical presentation of, predisposing factors in, and early and long-term outcome of patients treated surgically for intraperitoneal ruptured liver hydatid cysts. Medical records of 27 patients with traumatic rupture of hydatid cysts were evaluated retrospectively, as were records of 347 patients with nonperforated hydatid cysts. The ratio of perforation cases to nonperforation cases was 7.8%. Traffic accidents were the most common cause of perforation (n = 16). All patients had abdominal findings, and two patients (7%) had anaphylactic findings. The sensitivities of computed tomography and ultrasonography were 100% and 93%, respectively. Conservative surgical procedures were used for 80.5% of cysts and radical procedures for 19.5%. Associated organ injuries were determined in 10 patients. No significant difference was found between patients with peritoneal perforation and those without perforation in terms of sex (p = 0.403), previous hydatid disease surgery (p = 0.565), localization (p = 0.241), number of cysts (p = 0.537), presence of cystic content infection (p = 0.65), or presence of bile duct communication (p = 0.37). However, there were significant differences in age (p = 0.004), cyst diameter ( > 10 cm) (p = 0.03), and presence of superficially localized cysts (p = 0.011). Three patients developed recurrence. In the group of patients with perforation, the complication and recurrence rates were not statistically different in a comparison of surgical techniques (p = 0.37). No postoperative deaths occurred. The main predisposing factors for cyst perforation are young age and superficial localization. Peritoneal rupture increases the rates of postoperative morbidity and recurrence; in contrast, there was no significant relation between the operative procedure and the morbidity and recurrence rates. PMID- 17464538 TI - Staging laparoscopy for advanced gastric cancer: is it also useful for the group which has an aggressive surgical strategy? AB - BACKGROUND: Staging laparoscopy has been shown to be useful for increasing the accuracy of preoperative staging. However, controversy still exists regarding patient selection and subsequent treatment. The aim of this study was to determine the role of staging laparoscopy for a group that has a policy to perform aggressive surgery for advanced gastric cancer. METHODS: Twenty-four patients with clinical T3 or T4 gastric cancer expected to undergo curative resection, based on conventional preoperative diagnostic methods underwent staging laparoscopy. We examined the accuracy and the impact of staging laparoscopy on the further treatment options. RESULTS: The mean running time for the staging laparoscopy was 40.7 min (range: 25-75 min), and one complication was noted (4.2%). In regard to the tumor depth, 11 of 24 (45.8%) cases had a discrepancy after staging laparoscopy. In addition, 15 of 24 patients (62.5%) were found to have unsuspected peritoneal metastases, and 8 patients (33.3%) were excluded from laparotomy. The remaining 16 patients (66.7%), including 9 patients with localized peritoneal metastases (P1), underwent resection. The diagnostic accuracy for T factor was 81.3% in 16 laparotomy cases and overall accuracy of P factor was 91.7%. CONCLUSIONS: Staging laparoscopy had a significant impact on decisions regarding the treatment plan in patients with advanced gastric cancer for a group that has an aggressive treatment strategy. PMID- 17464539 TI - Cost/accuracy ratio analysis in breast cancer patients undergoing ultrasound guided fine-needle aspiration cytology, sentinel node biopsy, and frozen section of node. AB - We evaluated the effectiveness and the cost of axillary staging in breast cancer patients by ultrasound-guided fine-needle aspiration cytology (US-FNAC), sentinel node biopsy (SNB), and frozen sections of the sentinel node to achieve the target of the highest number of immediate axillary dissections. From January 2003 through October 2005, a total of 404 consecutive eligible breast cancer patients underwent US-FNAC of suspicious axillary lymph nodes. If tumor cells were found, immediate axillary dissection was proposed (33% of node-positive cases). If US or cytology was negative, SNB was performed. Frozen sections of the sentinel node allowed immediate axillary dissection in 31% of node-positive cases. The remaining 36% underwent delayed axillary dissection. We compared our policy with clinical evaluation of the axilla, showing better specificity of US-FNAC, the cost balanced by a 12% reduction of SNBs, and a marked reduction of unnecessary axillary dissections resulting from false-positive clinical staging. Moreover, the comparison between our policy and permanent histology of the sentinel node showed an 8% cost saving, mainly associated with the immediate axillary dissections. US-FNAC of axillary lymph nodes in breast cancer patients reliably predicts the presence of metastases and therefore refers a significant number of patients to the appropriate surgical treatment, avoiding an SNB. As cost saving to the health care system in our study is mainly related to one-step axillary surgery, US-FNAC of axillary lymph nodes and frozen section of the sentinel node generate significant cost saving for patients who have metastatic nodes. PMID- 17464540 TI - Male urogenital function after confirmed nerve-sparing total mesorectal excision with dissection in front of Denonvilliers' fascia. AB - This prospective study addresses the rate of male genital dysfunction following total mesorectal excision (TME) for rectal carcinoma using the anterior extramesorectal plane and its correlation with early urinary function, pelvic autonomic nerve preservation (PANP), and intraoperative neurostimulation (INS). A consecutive series of 44 men operated on by the same surgical team was analyzed. After excluding 18 patients considered to be impotent preoperatively, urogenital function was evaluated in 26 patients on the basis of the International Prostatic Symptom Score and International Index of Erectile Function. PANP was assessed with INS of parasympathetic nerves. PANP was complete in 21 patients (80.8%). Deterioration of urinary function was observed in six patients (23.1%) at early follow-up. Postoperative erectile dysfunction assessed in seven patients (26.9%) was associated with micturition disturbances in four (57%). Despite dissection in front of Denonvilliers fascia, the incidence of erectile dysfunction was low in patients with nonanterior tumors (1/10). INS results had higher sensitivity for predicting urinary dysfunction than for predicting erectile dysfunction (67% vs. 43%). Values for specificity and accuracy were 95% and 90%, and 89% and 77%, respectively. The correlation between the findings on INS and urinary function was good (kappa = 0.66) at a fair (kappa = 0.36) correlation for erectile function. Nerve-sparing TME using the anterior extramesorectal plane results in a justifiable rate of postoperative impotence in patients with nonanterior tumors. Patients with negative results on INS or early urinary dysfunction are at greater risk of erectile dysfunction. PMID- 17464541 TI - Association between implementation of clinical practice guidelines and outcome for traumatic brain injury. AB - BACKGROUND: Traumatic brain injury (TBI) is the leading cause of death in the population under 40 years of age in Western countries, and the same was true for Latvia in 1999. This indicated a strong need to improve the management of TBI. The Latvian Society of Neurosurgeons in collaboration with related societies created a dedicated working group, and the Guidelines for Medical Management of TBI in Latvia (Guidelines) were developed in 2001. This study aimed to assess the association between implementation of the Guidelines and the outcome of TBI patients. METHODS: The Guidelines were printed and distributed to relevant clinical units and teaching institutions. To assess the impact of the Guidelines on the outcome of TBI, Latvian medical statistics were researched. All patients admitted to emergency departments and registered as either discharged or dead with a diagnosis of head trauma from 1998 to 2004 were included in a retrospective survey. The primary endpoint accepted for analysis was the hospital case fatality rate (HCFR). RESULTS: The survey included 73,062 consecutive cases. The annual incidence rate of TBI admissions was stable during the period 1998 to 2004 (range 41.5-46.0/10,000), and the incidence of moderate and severe TBI (range 7.2-8.7/10,000) showed no significant trends. There was a reduction of HCFR from 3.7% during 1998-2000 to 2.6% during 2002-2004 (relative risk 0.72; 95% confidence interval 0.67-0.76; p = 0.03). CONCLUSION: Implementation of the Guidelines was associated with a statistically significant decrease of HCFR in TBI patients. PMID- 17464542 TI - The impact of technical factors on outcome of restorative proctocolectomy for familial adenomatous polyposis. AB - PURPOSE: This study was designed to assess the impact of technical factors on functional outcomes and complications in patients undergoing restorative proctocolectomy for familial adenomatous polyposis. METHODS: This was a descriptive study on 189 patients undergoing restorative proctocolectomy in a single tertiary referral center between 1977 and 2003. Primary outcomes were major complications, pouch function, and neoplastic transformation in the anal transitional zone. RESULTS: Pouch construction was J-reservoir (60 percent), W reservoir (34 percent), or S-reservoir (6 percent), with double-stapled (31 percent) or handsewn anastomosis with mucosectomy (69 percent). Overall pouch survival was 96 percent at five years and 89 percent at ten years, with no differences according to pouch design or anastomotic technique. The incidence of pelvic sepsis was unaffected by anastomotic technique (stapled vs. handsewn; 12 vs. 13 percent) or type of reservoir (J- vs. W- vs. S-pouch; 16 vs. 9 vs. 10 percent). Fistula formation was independent of anastomotic technique (stapled vs. handsewn; 8 vs. 8 percent) and type of reservoir (J- vs. W- vs. S-pouch; 9 vs. 7 vs. 0 percent). The night-time and 24-hour bowel frequencies were similar with the two anastomotic techniques and types of reservoirs. The incidence of polyps at the anal transitional zone was lower with handsewn than with stapled anastomosis (19 vs. 38 percent; P=0.047). CONCLUSIONS: Restorative proctocolectomy in patients with familial adenomatous polyposis has good functional outcomes and an acceptable rate of complications, which are independent of choice of technique. Handsewn ileoanal anastomosis with mucosectomy seems to reduce the incidence of subsequent neoplasia in the anal transitional zone but does not eliminate the risk of cancer. PMID- 17464543 TI - Learning curve for cytoreductive surgery and perioperative intraperitoneal chemotherapy for peritoneal surface malignancy--a journey to becoming a Nationally Funded Peritonectomy Center. AB - BACKGROUND: Cytoreductive surgery (CRS) combined with perioperative intraperitoneal chemotherapy (PIC) for peritoneal surface malignancy is associated with a morbidity rate of 30-50% and a mortality rate of 1-10%. Recently, the St George Hospital in Sydney has been commissioned as the Nationally Funded Center for treatment of peritoneal surface malignancy in Australia. METHODS: The clinical and treatment-related data regarding 140 consecutive patients were prospectively collected. A comparison between the initial 70 patients (Group I) and the subsequent 70 patients (Group II) was performed. Univariate and multivariate analyses were conducted to identify the significant risk factors for moderate to severe morbidity. RESULTS: The hospital mortality was 4%. Sixty-one patients (44%) had moderate morbidity. Twenty-eight patients (20%) experienced severe morbidity. The mean hospital stay was 30 days. Twenty-seven patients (19%) were readmitted after initial discharge for management of delayed complications. The severe morbidity rate reduced from 30% to 10%, and the delayed morbidity rate reduced from 29% to 10%, when comparing Groups I and II. There were also reduced transfusion requirement, duration of operation, and intensive care unit stay. In the multivariate analysis, Group I (vs Group II; P = .005), performing small bowel resection (P = .005), and >4 peritonectomy procedures (vs A in exon 3, and A181E in exon 4. No significant association with any of these variants was found, when compared to the frequencies of the variants in a healthy control cohort. Furthermore, the mutated alleles R20C and R72H did not segregate with the SLE phenotype in familial cases of SLE. Thus, our evaluation of the coding region of TNFRSF13b/TACI did not reveal any deleterious or disease-associated mutations. PMID- 17464556 TI - Triple site biventricular pacing in a patient with congestive heart failure and severe mechanical dyssynchrony. AB - We report on a case of a 62-year-old patient with symptomatic heart failure and severe ventricular electrical and mechanical dyssynchrony, who was implanted percutaneously with a triple site (dual-left single-right) resynchronization device. At 3-months follow-up, the patient's functional status improved significantly as shown by subjective and objective tests. Furthermore, this mode of pacing has allowed nearly complete inter- and intraventricular mechanical resynchronization. PMID- 17464557 TI - Intra-coronary guidewire mapping-a novel technique to guide ablation of human ventricular tachycardia. AB - HYPOTHESIS: Endocardial catheter ablation of ventricular tachycardia (VT) may fail if originating from epicardial or intramural locations. We hypothesized that mapping could be achieved using an angioplasty guidewire in the coronary circulation, to guide trans-coronary ablation. METHODS AND RESULTS: Six patients (2 male), 64 +/- 14 years and previously unsuccessful endocardial VT ablation were studied. Using ECG and existing endocardial mapping data, a coronary artery supplying the predicted VT origin was selected. A 0.014-in angioplasty guidewire was advanced into branches of the artery and connected to an amplifier to record unipolar signals against an indifferent electrode within the inferior vena cava. An uninflated angioplasty balloon was advanced over the wire such that only the distal 5 mm was used for mapping. One VT per patient was mapped (CL 348 +/- 102.1 ms). Diastolic potentials were recorded from all (77.7 +/- 43.8 ms pre-QRS onset) and concealed entrainment demonstrated in 3. Pacemapping during sinus rhythm was used in the remainder due to failure of entrainment (n = 2) or degeneration to VF (n = 1). Following branch identification, cold saline injection causing VT termination was used for further confirmation. Five VTs were ablated using intra coronary ethanol injection via the central lumen of the inflated over the wire balloon. The other was ablated using radiofrequency energy in a coronary vein adjacent to the target artery, which was too small for an angioplasty balloon. No complications or recurrence of ablated VT was seen over 19 +/- 17 months of follow up. CONCLUSIONS: Intracoronary guidewire mapping is a novel method of electrophysiological epicardial mapping to help guide trans-coronary VT ablation. PMID- 17464558 TI - What's in a name? Terms preferred by service recipients. AB - In a large (n = 1,827) multi-site study examining effectiveness of consumer operated service programs as an adjunct to traditional mental health services, we examined individuals' preferred term describing their status as service recipients, and we applied logistic regression to examine whether preference varied by gender, race or diagnosis. Preferred terms were client (39%), patient (22%), consumer (16%), survivor (11%), other (11%) and ex-patient (1%), varying by site. Controlling for site, preferences did not vary by gender, race, or diagnosis. The lack of consensus suggests clinicians, researchers, program administrators, and policymakers should be sensitive to individuals' preferences. PMID- 17464559 TI - Decreased total carbonic anhydrase esterase activity and decreased levels of carbonic anhydrase 1 isozyme in erythrocytes of type II diabetic patients. AB - In this exploratory study, we investigated total erythrocyte carbonic anhydrase (CA) estrase activity as well as CA I isozyme concentration in patients with diabetes mellitus type II (DM) and healthy individuals of Howard University Hospital community. Total estrase activity of CA was measured spectrophotometrically using p-nitrophenol acetate before and after inhibition with acetazolamide. CA I isozyme was measured by radial immunodiffusion using monoclonal antibody (CA I) in agarose plates. The study involved 20 consented participants; 10 normal (N) and 10 (DM), 21 to 84 years of age. The study was approved by the Howard University Institution Review Board. The CA activity was measured following lysis of cells as U/min/mL and CA I concentration as mg/l. We observed CA activity as 46.3+/-4(N) and 25+/-2.1 (DM) whereas CA I concentration as 1896+/-125 (N) and 1104 +/-63 (DM). We speculate that the change in the CA activity may of fundamental importance in the regulation of intracellular; pH(i) for the basic control of metabolism in diabetes mellitus. Further, we propose that CA activity is a good candidate for a biomarker of diabetes mellitus for the early detection of insulin resistance because the CA activity variation was proportional to the severity of the diabetes. PMID- 17464560 TI - Obituary: Abraham Rosenberg (1924-2006). PMID- 17464561 TI - One week of esomeprazole triple therapy vs 1 week of omeprazole triple therapy plus 3 weeks of omeprazole for duodenal ulcer healding in Helicobacter pylori positive patients. AB - In this randomized, double-blind, multicenter study, H. pylori-positive patients with an active duodenal ulcer (DU) received esomeprazole, 20 mg twice daily (bid), or omeprazole, 20 mg bid, with amoxicillin, 1000 mg bid, and clarithromycin, 500 mg bid, for 1 week (EAC and OAC, respectively). Patients received an additional 3 weeks of either placebo or omeprazole, 20 mg once daily (od), in the EAC and OAC groups, respectively. The intent-to-treat population included 374 patients (EAC, 186; OAC, 188). Four-week DU healing rates were similar in the EAC+placebo and OAC+omeprazole groups: 74% and 76%, respectively. DU healing rates at 8 weeks were 87% for EAC+placebo and 88% for OAC+omeprazole. H. pylori eradication rates were 75% and 79% for EAC and OAC, respectively. Both regimens were well tolerated. A 1-week regimen of esomeprazole-based H. pylori eradication triple therapy was as effective for DU healing and eradication of H. pylori as omeprazole-based triple therapy followed by an additional 3 weeks of monotherapy. PMID- 17464562 TI - Successful management of Aspergillus liver abscess in a patient with necrotizing fasciitis. PMID- 17464563 TI - Peptic ulcer in gastric heterotopia of the gallbladder without evidence of Helicobacter pylori infection. PMID- 17464564 TI - Liver fibrosis is associated with decreased peripheral platelet count in patients with chronic hepatitis B and C. AB - Thrombocytopenia is a common complication of chronic liver diseases, but its pathogenesis is not clear. Although generally attributed to hypersplenism, other factors should also be considered. We investigated the relationship between the peripheral platelet count and the degree of fibrosis in patients with chronic viral hepatitis. In an effort to avoid the effects of hypersplenism, we excluded patients with splenomegaly and/or bi- or pan-cytopenia. Seven hundred eighty-four patients (265 chronic viral hepatitis C and 519 chronic viral hepatitis B) were included in the study. Univariate analysis showed that the peripheral platelet count had a negative correlation with fibrosis score, necroinflammatory activity, and age in both groups. In multivariate analysis, the peripheral platelet count had a similar correlation with the fibrosis score and age, but not with necroinflammatory activity, in both groups. The peripheral platelet count decreased more significantly in females with chronic hepatitis C but not in the chronic hepatitis B group. In conclusion, a decrease in peripheral platelet count may be a sign of an increase in the degree of fibrosis during the course of chronic viral hepatitis B and C and factors other than hypersplenism may play a role in this decrease in the peripheral platelet count. PMID- 17464565 TI - Clinical use of functional magnetic resonance imaging: reflections on the new CPT codes. PMID- 17464566 TI - Molecular profiling in breast cancer. AB - Molecular profiling has provided biological evidence for the heterogeneity of breast cancer through the identification of intrinsic subtypes like Luminal A, Luminal B, HER2+/ER- and basal-like. It has also led to the development of clinically applicable gene expression-based prognostic panels like the Mammaprint and Oncotype Dx. The increasingly sophisticated understanding allowed by this and similar technology promises future individualized therapy. PMID- 17464567 TI - Urban poverty: an urgent public health issue. PMID- 17464568 TI - Emerging strategies for healthy urban governance. AB - Urban health promotion is not simply a matter of the right interventions, or even the necessary resources. Urban (and indeed global) health depends to an important extent on governance, the institutions and processes through which societies manage the course of events. This paper describes the concept of governance, distinguishing between reforms aimed at improving how government works and innovations that more fundamentally reinvent governance by developing new institutions and processes of local stakeholder control. The paper highlights strategies urban governors can use to maximize their influence on the national and international decisions that structure urban life. It concludes with some observations on the limitations of local governance strategies and the importance of establishing a "virtuous circuit" of governance through which urban dwellers play a greater role in the formation and implementation of policy at the national and global levels. PMID- 17464569 TI - Community integration of transition-age individuals: views of young with mental health disorders. AB - This qualitative study examines the perceptions of young adults with mental health disorders of community integration. Fifty-nine young men and women participated in 12 focus groups whose aim was to gain understanding of what community integration means to them. Focus group questions also explored barriers and supports for their community integration, as well as their goals for the future and advice to others facing similar challenges. Themes that emerged were reported within the multiple domains that participants used to describe their experiences of community integration (or the lack thereof). This study highlights the desires of these young people to achieve goals in education and employment and to have friendships. Participants identified a pervasive lack of understanding of mental health and prevalent stigmatizing attitudes as resulting in challenges to their community integration. Implications of the study discuss roles for behavioral health services in encouraging empowerment, choices, and connections so that young people with mental health disorders may achieve their preferred levels of community integration. PMID- 17464570 TI - A single-quantum methyl 13C-relaxation dispersion experiment with improved sensitivity. AB - A pulse sequence is described for recording single-quantum (13)C-methyl relaxation dispersion profiles of (13)C-selectively labeled methyl groups in proteins that offers significant improvements in sensitivity relative to existing approaches where initial magnetization derives from (13)C polarization. Sensitivity gains in the new experiment are achieved by making use of polarization from (1)H spins and (1)H --> (13)C --> (1)H type magnetization transfers. Its utility has been established by applications involving three different protein systems ranging in molecular weight from 8 to 28 kDa, produced using a number of different selective labeling approaches. In all cases exchange parameters from both (13)C-->(1)H and (1)H --> (13)C --> (1)H classes of experiment are in good agreement, with gains in sensitivity of between 1.7 and 4 fold realized using the new scheme. PMID- 17464571 TI - Information provided by generic and brand-name pharmaceutical manufacturers in response to a request. AB - AIM: To assess the medical information provided by manufacturers in response to a specific request, and to compare the responses between generic and brand-name companies. SETTING: Community pharmacy in Spain. METHOD: A systematic request for product monographs was made between 1999 and 2002 to manufacturers registering new medicines in Spain. A standardised letter was sent to the medical affairs departments. If there was no reply after 3 months, a second standardised letter was sent requesting the monograph. Blood derivatives, intravenous medicines, and radiological contrast agents were excluded. MAIN OUTCOME MEASURES: The delay that occurred in receiving information and the type of material sent in response to the request was compared between the two types of companies. RESULTS: About of 833 medicines from 185 manufacturers were registered during the time period studied. After applying exclusion criteria, 805 medicines, including 419 (52.0%) generic and 386 (48.0%) brand-name products, were analyzed. No replies were received for 242 (30.0%) requests 183 (43.7%) generics and 59 (15.3%) brand names; P < 0.005). We received 369 (65.5% of 533) replies after the first request: 140 of 236 (59.3%) generics and 229 of 327 (70.0%) brand-names (P = 0.009). The average response delay was 9.7 days [CI95%: 8.65-10.68]. There was a statistically significant difference between generic and brand-name companies after the first request (P = 0.001), but not after the second request (P = 0.312). CONCLUSION: Brand-name manufacturers reply more often, more quickly, and with better quality information than generic manufacturers. PMID- 17464572 TI - Utilization of antimicrobial agents with and without prescription by out-patients in selected pharmacies in South-eastern Nigeria. AB - OBJECTIVE: We conducted a study in out-patient pharmacies in South-eastern Nigeria in order to determine the extent of self-medication of antimicrobial agents in this area, assess the dosing error associated with this practice and to ascertain the extent of involvement of community pharmacies. METHOD: A survey was carried out daily in selected community pharmacies for a period of 90 days. Data were collected on the number of patients visiting these shops for antimicrobial agents, the number getting their medication with a prescription, the number getting their order without prescription and on the type and dose of antimicrobial agents received. MAIN OUT-COME MEASURE: The percentage of patients with prescription and without prescription was compared. The percentage under dosages or over-dosages associated with how each antimicrobial agent was obtained were compared. The total DDDs of antimicrobial agents dispensed within this period with and without prescription were compared. RESULTS: A total of 4,128 outpatients visited the shops for antimicrobial agents within the period and were involved in the study. Of this number, 1,742 (42.2%) came with a prescription from qualified medical personnel and 2,386 (57.8%) came without a prescription. A total of 13,693.13 DDDs of antimicrobial agents was dispensed, of which 56.38% was dispensed with prescription and 43.62% was dispensed without prescription. The degrees of under-dosing were significantly (P < 0.05) higher in regimen filled without prescription when compared to those filled with prescription. CONCLUSION: Majority of the patients in this region still obtains their antimicrobial agents without a proper prescription, which is associated with sub therapeutic dosing of these agents. There is a need for better regulation of antimicrobial agents dispensing and utilization in the region. PMID- 17464573 TI - Narratives about illness and medication: a neglected theme/new methodology within pharmacy practice research. Part II: medication narratives in practice. AB - AIM: Part 2 of this paper aims to provide a methodological framework for the study of medication narratives, including a semi-structured interview guide and suggested method of analysis, in an attempt to aid the development of narrative scholarship within pharmacy practice research. Examples of medication narratives are provided to illustrate their diversity and usefulness. METHODS: The framework is derived from the work of other researchers and adapted for our specific purpose. It comes from social psychology, narrative psychology, narrative anthropology, sociology and critical theory and fits within the social constructionist paradigm. The suggested methods of analysis could broadly be described as narrative analysis and discourse analysis. Examples of medication narratives are chosen from a variety of sources and brief interpretations are presented by way of illustration. CONCLUSION: Narrative analysis, a neglected area of research in pharmacy practice, has the potential to provide new understanding about how people relate to their medicines, how pharmacists are engaged in producing narratives and the importance of narrative in the education of students. IMPACT OF THE ARTICLE: This article aims to have the following impact on pharmacy practice research: Innovative approach to researching and conceptualising the use of medicines. Introduction of a new theoretical perspective and methodology. Incorporation of social science research methods into pharmacy practice research. Development of narrative scholarship within pharmacy. PMID- 17464574 TI - The role of plant mitochondria in the biosynthesis of coenzymes. AB - This last decade, many efforts were undertaken to understand how coenzymes, including vitamins, are synthesized in plants. Surprisingly, these metabolic pathways were often "quartered" between different compartments of the plant cell. Among these compartments, mitochondria often appear to have a key role, catalyzing one or several steps in these pathways. In the present review we will illustrate these new and important biosynthetic functions found in plant mitochondria by describing the most recent findings about the synthesis of two vitamins (folate and biotin) and one non-vitamin coenzyme (lipoate). The complexity of these metabolic routes raise intriguing questions, such as how the intermediate metabolites and the end-product coenzymes are exchanged between the various cellular territories, or what are the physiological reasons, if any, for such compartmentalization. PMID- 17464575 TI - Negative ERGs in mucopolysaccharidoses (MPS) Hurler-Scheie (I-H/S) and Hurler (I H)-syndromes. AB - The configuration and progression of the ERG in two children with mucopolysaccharidosis (MPS) I H/S (Hurler-Scheie syndrome) and MPS I H (Hurler syndrome) is described. Physical examination, biochemical analysis, ophthalmic examination and electroretinography were performed. The Hurler-Scheie patient (case 1) showed negative scotopic but normal photopic ERGs, which remained unchanged over 2 years. The Hurler patient (case 2) showed negative scotopic and photopic ERGs which did not alter after bone marrow transplantation (BMT). One year after BMT, further b-wave amplitude reduction had caused the ERGs to become more negative. The electronegative configuration of the ERGs suggests that, in these cases of MPS, the primary retinal abnormality in MPS I may be faulty synaptic transmission from photoreceptors to more proximal elements, deficient bipolar responsivity, or Muller cell disease. Further degradation with time suggests the defect to be progressive with BMT causing little or no improvement. In the Hurler-Scheie syndrome case, the defect appears to spare the cone system and to show little or no progression. PMID- 17464577 TI - Effect of passive uterine straightening during embryo transfer: a systematic review and meta-analysis. AB - BACKGROUND: Part of the success of ultrasound-guided embryo transfer has been associated with the beneficial effect of uterine straightening by passive bladder distention. Even so, this has not been properly analysed in the literature. METHODS: This is a systematic review and meta-analysis of prospective, randomised, controlled trials, comparing embryo transfer with a full versus empty bladder. Electronic (e.g. PubMed, EMBASE, Cochrane Library) and hand searches were performed to locate trials. Primary outcomes were live-birth, ongoing and clinical pregnancy rates. Secondary outcomes were rates of implantation, miscarriage, multiple and ectopic pregnancies, and retained embryos. Also, the ease of transfer, need for instrumental assistance, and presence of blood on the catheter tip were evaluated. Four studies were identified, of which 1 study was excluded. Meta-analysis was conducted with the Mantel-Haenszel method, utilising the fixed-effect model. RESULTS: For the primary outcome measures, no data was available for the LBR rate. There was a significantly higher chance of an ongoing pregnancy [OR=1.44 (95% CI=1.04-2.04)] and clinical pregnancy [OR=1.55 (95% CI=1.16-2.08)] with a full bladder. For the secondary outcomes, there was a significantly greater incidence of difficulty, or need for instrumental assistance, with an empty bladder. Other outcome measures were not significantly different. CONCLUSION: There is evidence in the literature advising to fill the bladder prior to embryo transfer. PMID- 17464578 TI - Effect of maternal herpes simplex virus (HSV) serostatus and HSV type on risk of neonatal herpes. AB - BACKGROUND: Neonatal herpes simplex virus (HSV) is a rare but devastating disease. We have conducted pooled analyses of data from 3 cohorts to evaluate the effects of maternal HSV serostatus and HSV type on risk of neonatal HSV acquisition and severity. METHODS: Data from cohorts in Seattle, WA, and Stanford, CA, USA, and Stockholm, Sweden were pooled using Mantel-Haenszel methods. RESULTS: Seventy-eight infants with documented neonatal HSV and known maternal HSV serostatus were included. The risk of neonatal HSV-2 infection was similar in infants born to HSV seronegative women compared with HSV-1 seropositive women (pooled OR: 1.6; 95% CI: 0.6-4.0). The odds of neonatal HSV infection was increased in the presence of exposure to maternal HSV-1 versus HSV 2 (adjusted pooled OR: 19.2; 95% CI: 5.8-63.6). An elevated odds of disseminated HSV in infants born to women with newly acquired genital herpes was observed in Stockholm (OR=13.5; 95% CI: 1.4-630), but not in Seattle or Stanford. CONCLUSION: Our results suggest that maternal HSV-1 antibody offers little, if any, protection against neonatal HSV-2 infection. During reactivation, HSV-1 appears more readily transmissible to the neonate than HSV-2, a concerning finding given the rising frequency of genital HSV-1 infection. PMID- 17464579 TI - Screening for pre-eclampsia and small for gestational age fetuses at the 11-14 weeks scan by uterine artery Dopplers. AB - OBJECTIVE: To assess the role of uterine artery Doppler studies at 11-14 weeks in screening for pre-eclampsia (PET), small for gestational age (SGA) fetuses, and placental abruption. METHODS: Prospective study on 1,123 women presenting for routine ultrasound examination at 11-14 weeks for nuchal translucency measurement. Uterine artery blood flow was studied by transvaginal colour Doppler, the mean pulsatility index (PI) was calculated, and the presence of a diastolic notch was recorded. RESULTS: The mean, median and 95th centile of uterine artery PI were 1.71, 1.64 and 2.54, respectively. Bilateral notches were observed in 63.4%, and a unilateral notch in 18.4% of cases. The sensitivity of mean uterine artery PI>or=95th centile for PET, early onset severe PET necessitating delivery before 34 weeks, SGAor=4 h (watching television and reading) daily; 19% were defined as non-exercisers before pregnancy, 30% in the first trimester, 36% in the second trimester and 53% in the third trimester. Fifty women (10.6%) continued to exercise>or=4 times/week in the third trimester. Mean weight gain was 13.8 kg (SD: 5.2). Of the normal pregestational weight women (pre-BMI: <26) and overweight women (pre-BMI: >or=26), 32 and 51% had exceeded weight gain above accepted recommendations, respectively. Women who exercised regularly had significantly lower weight gain than inactive women in the third trimester only. CONCLUSION: Pregnant women have a low total physical activity level, and a high percentage of women exceed the recommended weight gain during pregnancy. PMID- 17464585 TI - Risk of preterm delivery in relation to maternal low birth weight. AB - OBJECTIVE: We examined the relationship between maternal low birth weight and preterm delivery risk. METHODS: Information concerning maternal birth weight was collected during in-person interviews. Logistic regression was used to estimate odds ratios (OR) and 95% confidence intervals (95% CI). Preterm delivery cases were studied in aggregate, in subgroups (spontaneous preterm labor, preterm premature rupture of membranes, medically induced preterm delivery, moderate preterm delivery [gestational age at delivery 34-36 weeks], and early preterm delivery [gestational age at delivery<34 weeks]). RESULTS: After adjusting for confounders, women weighing<2,500 g at birth had a 1.54-fold increased risk of preterm delivery versus women weighing=2,500 g (95% CI 0.97-2.44). Maternal low birth weight was associated with a 2-fold increased risk of spontaneous preterm delivery (95% CI 1.03-3.89), but weakly associated with preterm premature rupture of membranes (OR=1.44; 95% CI 0.67-3.09) and medically induced preterm delivery (OR=1.10; 95% CI 0.43-2.82). Maternal low birth weight was more strongly associated with early preterm delivery (OR=1.94) than with moderate preterm delivery (OR=1.46). Women weighing<2,500 g at birth and who became obese (pre pregnancy body mass index, =30 kg/m2) before pregnancy had a 3.65-fold increased risk of preterm delivery (95% CI 1.33-10.02) versus women weighing=2,500 g at birth and who were not obese prior to pregnancy (<30 kg/m2). CONCLUSIONS: Results confirm earlier findings linking maternal low birth weight with future risk of preterm delivery. PMID- 17464586 TI - Rupture of the scarred uterus. AB - OBJECTIVE: To assess the risk of uterine rupture of the scarred uterus according to mode of delivery in subsequent births recorded as spontaneous labour, labour induced by oxytocin, labour after ripening with prostaglandin E2, and planned cesarean section. METHODS: Retrospective study of 2,128 births with a low transversal scar after a previous cesarean section. The study population was realised in a level III university hospital from 1995 to 2003. The association between mode of delivery and uterine rupture was studied in a multivariate logistic regression model, and adjusted for specific antenatal confounding factors. RESULTS: Over 9 years, we collected 22 cases (1%), including 11 asymptomatic ruptures in a population of 2,128 scarred uteri out of 28,248 deliveries. Uterine rupture occurred at a rate of 0.3 per 100 among women with repeated cesarean delivery without labour, 1 per 100 among women with spontaneous onset of labour, 1.4 per 100 among women with oxytocin-induced labour, and 2.2 per 100 among women with prostaglandin cervical ripening. Compared to women with a planned cesarean section, women with spontaneous onset of labour were more likely to have uterine rupture (OR: 4.0; 95% CI: 0.8-42.0). A greater relative risk was observed among women with oxytocin-induced labour (OR: 4.3; 95% CI: 0.3 60.0), and particularly those with prostaglandin-induced labour (OR: 8.7; 95% CI: 1.5-97.3, p=0.01). CONCLUSION: In women with a scarred uterus, prostaglandin E2 induction of labour is a risk factor for uterine rupture. The practice of a systematic cesarean section in cases with Bishop score<3, appropriate induction procedure, and rigorous monitoring of the labour, could make for a safer delivery. PMID- 17464587 TI - New insights into fatigue and health-related quality of life after delivery. AB - BACKGROUND: A delivery has a major impact on the health-related quality of life (HRQoL) of the new mother, especially on fatigue. A common complication during delivery that might have a relationship with maternal morbidity is blood loss. The objectives were to investigate fatigue and HRQoL in women after vaginal delivery (VD), elective caesarean section (CS) and emergency CS, and its relationship with postpartum hemoglobin (Hb) levels during the first 6 weeks postpartum. METHODS: Some 141 patients (71 after VD, 36 after elective CS and 34 after emergency CS) completed the HRQoL questionnaires MFI and EQ-5D between 12 and 24 h after VD and 24-48 h after CS (t=0). At 1, 3 and 6 weeks postpartum these questionnaires were repeated, together with the SF36. RESULTS: Patients after VD had higher mean physical HRQoL scores than after CS. The average period to reach full physical recovery was 3 weeks after VD, 6 weeks after elective CS, and >6 weeks after emergency CS. Mean mental HRQoL scores of the study groups were similar or even better compared to reference values. The significant correlation between Hb level and mean physical HRQoL scores found at t=0 had disappeared at 1 week postpartum. CONCLUSIONS: Results of this study provided insights into the natural course of fatigue and HRQoL postpartum. Important differences in fatigue and HRQoL scores were observed between the 3 modes of delivery. These HRQoL measures can be used in future clinical trials to assess the effects of interventions postpartum. PMID- 17464588 TI - Sequence variants in exons of the BMP-15 gene in Chinese patients with premature ovarian failure. AB - BACKGROUND: The bone morphogenetic protein-15 (BMP-15) gene is thought to be one of the important candidate genes involved in premature ovarian failure (POF). However, to date, there has been no consensus on the relationship between mutations in BMP-15 and ovarian dysfunction. OBJECTIVE: To analyse mutations in the BMP-15 gene in Chinese women with POF. MATERIALS AND METHODS: We sequenced the BMP-15 gene protein coding region of 92 patients and 76 healthy controls that cycle regularly. RESULTS: No mutations were found in the BMP-15 gene protein coding region. There was no difference in the incidence of single nucleotide polymorphism (SNP) rs 17003221 (CT) in exon 2 (p>0.05), or SNP rs (3810682CG: ss16336587) in the putative promoter region of exon 1, between the two groups. However, the allele gene frequency in SNP rs (3810682CG: ss16336587) was C (97.92%) and G (2.08%), respectively, in Chinese women, which is different from other races. CONCLUSIONS: Our findings indicate that mutations in BMP-15 exons, or changes in BMP-15 pro-peptide, are rare in Chinese women with POF. In addition, our data suggest that the 2 SNPs are not related to POF in Chinese women, while providing evidence for SNP variation between different races. PMID- 17464589 TI - Young women's sexual health and their views on dialogue with health professionals. AB - BACKGROUND: Many women who experience sexual problems or who have been subjected to sexual abuse, do not seek help, and often health professionals avoid raising such issues. The aim of this study was to describe young women's sexual health and their views on a dialogue about aspects of sexuality in connection with a gynecologic examination. METHOD: Cross-sectional study. Women, 23, 26 and 29 years of age, called for gynecologic screening between March and July 2006 (n=488; response rate 75%). Descriptive statistics, multiple logistic regression and bivariate analysis were used. RESULTS: The majority (95%) had a positive attitude towards sexuality, although many women reported sexual problems, and one fifth (22%) were dissatisfied with their sex life. Women considered it appropriate to be asked about sexuality in general (92%), while a majority (72%) found questions about sexual abuse appropriate. However, depending on the area, the majority (76-99%) had never been asked such questions. Demographic variables, aspects of life, sexuality and sexual abuse were irrelevant for whether women considered it appropriate to be asked questions about aspects of sexuality, by a midwife/doctor in connection with a gynecologic examination. CONCLUSION: The result can activate health professionals to initiate a dialogue about sexual issues among young women. In this way, many women who experience sexual problems or have been subjected to sexual abuse, may be provided the opportunity to improve their sexual health. PMID- 17464590 TI - Loop electrosurgical excision of the cervix and the subsequent risk of preterm delivery. AB - OBJECTIVES: To investigate the association between loop electrosurgical excision procedure (LEEP) and other potential risk factors, and subsequent preterm delivery (<37 weeks), using data from a large cohort study of Danish women. METHODS: The Danish prospective cohort study was initiated in 1991, with the original aim of investigating the role of human papillomavirus in the natural history of cervical neoplasia. The study included 11,088 women aged 20-29. The cohort was invited for 2 follow-up examinations in 1993-1995 and 1999-2000, respectively. At all 3 examinations, the women answered questions about a number of different lifestyle variables. We assessed the relationship between preterm delivery and potential risk factors, such as previous LEEP treatments, smoking during pregnancy, age, parity and previous preterm delivery. The cohort was followed until 2004, through linkages with the nationwide Pathology Data Bank and the Medical Birth Registry. RESULTS: Of the 14,982 deliveries in the cohort during follow up, 542 were preterm (21-37 weeks). Among deliveries with no previous LEEP, 3.5% ended as a preterm delivery, whereas this applied to 6.6% among deliveries following a LEEP, yielding a significantly increased risk of preterm birth after LEEP (OR=1.8; 95% CI: 1.1-2.9). The strongest risk factor for preterm delivery was a previous preterm delivery (OR=2.3; 95% CI: 1.4-3.7). Other significant risk factors were smoking during pregnancy and low educational status. CONCLUSION: Our study showed an almost 2-fold increase in the risk of preterm delivery after LEEP treatment. Thus, women in their reproductive age should be informed about the increased risk of preterm delivery, if treated with LEEP. PMID- 17464591 TI - Medical management of first trimester miscarriage according to ultrasonographic findings. AB - BACKGROUND: The efficacy of medical treatment of first trimester miscarriages may depend on the regimen used, the definition of success, clinical symptoms, and, possibly, on the ultrasonographic findings. Our primary aim was to assess if a single dose of misoprostol could reduce the number of surgical interventions in early pregnancy failure, and to compare efficacy in different ultrasonographically-defined subgroups. METHODS: A retrospective review of 355 consecutive patients treated for a first trimester miscarriage with a single dose of vaginal misoprostol (400 microg), followed by vaginal ultrasonography after either 1, 2 or 3 days. Treatment was successful if a complete abortion was diagnosed at follow-up. The women were divided into 4 ultrasonographically defined groups: missed abortion with a crown rump length (CRL)>or=6 mm (Group A1) or CRL<6 mm (Group A2), anembryonic gestation with gestational sac>or=18 mm (Group B1) or <18 mm (Group B2). RESULTS: The overall success rate was 39.2% (139/355). The regimen was more efficacious in Group A2 (50%) and less efficacious in Group B1 (26.6%) than in the other groups. The success rates were lower 1 day after treatment (30.2%) compared to days 2 and 3 (43.6%; p<0.05), and the difference was largest in Group B1 (12.9 versus 35.4%; p<0.05). CONCLUSIONS: Single dose misoprostol reduced the number of surgical interventions. The success rates of medically-treated first trimester miscarriages varied according to the ultrasonographic definitions of pregnancy failure, time of assessment, and the criteria for success. PMID- 17464592 TI - Surgical staging, treatment, and follow-up of borderline tumors in different hospital categories: a prospective nationwide survey in Finland. AB - BACKGROUND: Surgical treatment and staging of ovarian borderline tumors have been reported to be often suboptimal and differ considerably. We evaluated the extent of surgical treatment of these tumors in different hospital categories. MATERIAL AND METHODS: A prospective survey performed in 1999 included 65 patients operated on for borderline ovarian tumors and covered 78% of such patients reported to the Finnish Cancer Registry. Detailed information of demographic data and surgical treatment was reported by the responsible physicians using a special questionnaire after confirmation of histopathology. RESULTS: Fifty-eight patients (89%) had stage I tumor, only two patients (3%) had stage II disease and five patients (8%) had stage III disease with peritoneal implants. The majority of the patients underwent bilateral salpingo-oophorectomy (66%) and hysterectomy (58%). Unilateral salpingo-oophorectomy was performed for 21 (32%) and omentectomy for 22 (34%) patients. Ten out of the 16 women under 40 years of age had fertility sparing surgery. Peritoneal biopsies were taken in 16 (25%) women and lymphadenectomy was performed for 9 (14%) patients with clinical suspicion of invasive ovarian carcinoma. Frozen section was taken in half of the patients and the histology remained the same in 72% of the final pathology reports. No clear differences of the extent of surgical treatment were detected between different hospital categories. Overall cumulative 5-year relative survival rate was 96%. CONCLUSIONS: Bilateral salpingo-oophorectomy and hysterectomy was performed for the majority of patients with borderline ovarian tumor. More attention should be paid to adequate staging of borderline tumors in all hospital categories. PMID- 17464593 TI - Sentinel node and vulvar cancer: a series of 47 patients. AB - BACKGROUND: There is growing interest to apply the sentinel node technique in the treatment of vulvar cancer. METHODS: All charts of the patients operated on for vulvar cancer at Tampere University Hospital from January 1, 2001 through June 30, 2005 were retrospectively reviewed. Demographic, clinical, and histopathological information was collected from each patient. The sentinel lymph node mapping was done intraoperatively either with a combination of the radioisotope and dye techniques (40 patients) or with the dye technique alone (7 patients). The sentinel lymph node was dissected separately for histopathological evaluation, and then a routine inguinal lymphadenectomy was performed. RESULTS: The final FIGO surgical Stage distribution was: Stage I, 11 (23%); Stage II, 14 (30%); Stage III, 21 (45%); and Stage IV, 1 (2%). Sentinel lymph node was identified in 46 (98%) women with either one or both of the methods. In Stage I II, the sentinel lymph node identification rate was 25/25 (100%) with the combined method. The only patient with unidentified sentinel lymph node had lymphatic spread beyond inguinal area or Stage IV disease. Eighteen of the sentinel lymph nodes (39%) were positive for tumor cells, and in 5 cases additional metastatic nodes were found. One patient with macroscopically enlarged metastatic inguinal nodes and Stage III disease had a negative sentinel lymph node. In the 25 patients with Stage I-II disease, the false-negative rate of the sentinel lymph node method was 0/4, giving a negative predictive value of 1.00. CONCLUSIONS: A sentinel node identification rate of 98% with a false-negative rate of 0% in the patients with Stage I-II disease is an encouraging finding. PMID- 17464594 TI - Surgery of borderline tumors of the ovary: retrospective comparison of short-term outcome after laparoscopy or laparotomy. AB - BACKGROUND: Laparoscopic management of borderline ovarian tumors is controversial. OBJECTIVE: To retrospectively compare outcome after surgery by laparoscopy or laparotomy for borderline tumors. METHODS: Ovarian tumors from all women operated at Ulleval University Hospital during a five-year period were re evaluated histologically. Patients with borderline FIGO (International Federation of Gynaecology and Obstetrics) stage I tumors were retrospectively compared regarding surgery outcome following laparoscopy or laparotomy. RESULTS: Histological re-evaluation revealed only 3 misclassifications in 608 patients. Borderline tumors represented 36% of epithelial ovarian malignancies. The 107 borderline stage I included 52 serous, 53 mucinous, and 2 endometrioid tumors. Thirty-eight patients were operated on primarily by laparoscopy and 69 by laparotomy (including 14 women starting with laparoscopy). In the laparoscopy group, more women were premenopausal (63% versus 35%, p=0.01) and median tumor diameter was smaller (8.6 versus 16.4 cm, p<0.001) as compared to the laparotomy group. When tumor diameter exceeded 10 cm, intraoperative tumor rupture was significantly more frequent during laparoscopy than during laparotomy (p=0.01). Less postoperative complications were seen after laparoscopic operations (p=0.034), but laparoscopic surgeries were less extensive, without hysterectomy, as compared to laparotomy. During the 14-78 months follow-up time, no relapse occurred in either group. After fertility-sparing surgery, there was no statistical significant difference regarding successful pregnancies between the two groups. CONCLUSIONS: Laparoscopic treatment of borderline ovarian tumors is feasible if tumor is of moderate size (diameter below 10 cm), gives fewer complications, and shorter hospital stay. Long-term follow-up of larger materials is needed to determine the ultimate recurrence risk as well as fertility rates. PMID- 17464595 TI - Place of delivery among women who had antenatal care in a teaching hospital. AB - BACKGROUND: The utilization of antenatal care services does not necessarily equate to delivery at the health facility. The objectives of this study were to determine the proportion of pregnant women who had antenatal care and delivered in the hospital and the reasons for delivery elsewhere. MATERIALS AND METHODS: Pregnant women of low risk who initiated antenatal care at the University Teaching Hospital were recruited consecutively from January to June 2004 and longitudinally followed up until delivery. A two-part questionnaire was used to obtain information on biosocial data, place of delivery, accoucher, fetomaternal outcome, and reason(s) for delivery elsewhere. RESULTS: Of the 1,080 women studied, 740 (68.5%) delivered in the hospital while 340 (31.5%) delivered elsewhere. The majority (52) of the deliveries elsewhere were at the woman's home. There were more literate women amongst the hospital delivery group, while there were more women of high parity and with a previous home birth in the home delivery group (p<0.05). Fetomaternal outcomes were similar in both groups. Nurses/midwives conducted most (60%) of the home deliveries and the principal reasons for home births were privacy and lack of transport during labor. CONCLUSION: 68.5% of women who had uneventful antenatal care delivered in the hospital. Home delivery was the commonest site for delivery elsewhere and the main reasons were privacy and lack of transportation. Skilled attendants conducted most of the home deliveries with good fetomaternal outcome. There might be a need to encourage women during antenatal care to reach out for skilled attendants when home delivery becomes inevitable. PMID- 17464596 TI - Postmenopausal vaginal bleeding caused by endometrial tuberculosis. PMID- 17464597 TI - Incomplete mole with a false-low level of human chorionic gonadotropin and hyperthyroidism. PMID- 17464598 TI - A triplet pregnancy featuring a 47, XY+9 heterozygous complete mole coexisting with two fetuses at 9 weeks. PMID- 17464599 TI - Reliability of cord blood analysis at birth. PMID- 17464601 TI - High-dose local infiltration analgesia after hip and knee replacement--what is it, why does it work, and what are the future challenges? PMID- 17464602 TI - Don't be misled by the orthopedic literature : tips for critical appraisal. PMID- 17464603 TI - Comparison of peri- and intraarticular analgesia with femoral nerve block after total knee arthroplasty: a randomized clinical trial. AB - BACKGROUND: Postoperative pain after total knee arthroplasty (TKA) can be difficult to manage and may delay recovery. Recent studies have suggested that periarticular infiltration with local anesthetics may improve outcome. METHODS: 80 patients undergoing TKA under spinal anesthesia were randomized to receive continuous femoral nerve block (group F) or peri- and intraarticular infiltration and injection (group I). Group I received a solution of 300 mg ropivacaine, 30 mg ketorolac, and 0.5 mg epinephrine by infiltration of the knee at the end of surgery, and 2 postoperative injections of these substances through an intraarticular catheter. RESULTS: More patients in group I than in group F could walk < 3 m on the first postoperative day (29/39 vs. 7/37, p < 0.001). Group I also had significantly lower pain scores during activity and lower consumption of opioids on the first postoperative day. No differences between groups were seen regarding side effects or length of stay. INTERPRETATION: Peri- and intraarticular application of analgesics by infiltration and bolus injections can improve early analgesia and mobilization for patients undergoing TKA. Further studies of optimal drugs, dosage, and duration of this treatment are warranted. PMID- 17464604 TI - Reduced hospital stay and narcotic consumption, and improved mobilization with local and intraarticular infiltration after hip arthroplasty: a randomized clinical trial of an intraarticular technique versus epidural infusion in 80 patients. AB - BACKGROUND: Epidural analgesia gives excellent pain relief but is associated with substantial side effects. We compared wound infiltration combined with intraarticular injection of local anesthetics for pain relief after total hip arthroplasty (THA) with the well-established practice of epidural infusion. METHODS: 80 patients undergoing elective THA under spinal block were randomly assigned to receive either (1) continuous epidural infusion (group E) or (2) infiltration around the hip joint with a mixture of 100 mL ropivacaine 2 mg/mL, 1 mL ketorolac 30 mg/mL, and 1 mL epinephrine 0.5 mg/mL at the conclusion of surgery combined with one postoperative intraarticular injection of the same substances through an intraarticular catheter (group A). RESULTS: Narcotic consumption was significantly reduced in group A compared to group E (p = 0.004). Pain levels at rest and during mobilization were similar in both groups but significantly reduced in group A after cessation of treatment. Length of stay was reduced by 2 days (36%) in group A compared to group E (p < 0.001). INTERPRETATION: Wound infiltration combined with 1 intraarticular injection can be recommended for patients undergoing THA. Further studies of dosage (high/low) and duration of intraarticular treatment are warranted. PMID- 17464605 TI - Postoperative analgesia in total hip arthroplasty: a randomized double-blinded, placebo-controlled study on peroperative and postoperative ropivacaine, ketorolac, and adrenaline wound infiltration. AB - BACKGROUND: Comfort and lack of pain are important for optimal mobilization after hip replacement. We investigated the efficacy of double wound infiltration. PATIENTS AND METHODS: 40 consecutive patients undergoing total hip replacement were randomized into two groups in this double-blinded study. They received wound infiltration at the end of surgery and through an intraarticular catheter 24 h postoperatively. The catheter was placed at the end of surgery. One group received solutions of ropivacaine, ketorolac, and adrenaline. Patients in the control group were injected with saline instead. The observation period was 6 weeks. RESULTS: The patients who received the analgesic solution had less pain up to 2 weeks postoperatively. They reached an earlier and lower pain minimum during the first days postoperatively, had lower use of analgesia up to day 4 postoperatively, and were more satisfied. Use of analgesic solution resulted in less joint stiffness and better function 1 week postoperatively. INTERPRETATION: Operative and postoperative wound infiltration with multimodal drugs reduces pain and the requirement for analgesics after hip replacement, leading to faster postoperative mobilization. PMID- 17464606 TI - Continuous psoas and sciatic block after knee arthroplasty: good effects compared to epidural analgesia or i.v. opioid analgesia: a prospective study of 63 patients. AB - INTRODUCTION: For endoprosthetic knee surgery, intensive postoperative pain therapy is necessary. We therefore evaluated whether the combination of continuous psoas compartment and sciatic analgesia (PSC) is as effective as epidural analgesia (EPI) and whether it provides better analgesia than patient controlled intravenous analgesia with piritramide (PCA). METHODS: We studied 63 patients who underwent total knee arthroplasty (TKA). The PSC group received a combination of continuous psoas and sciatic nerve block, the EPI group an epidural analgesia, and the PCA group an intravenous patient-controlled piritramide pump. Pain scores, satisfaction, flexion and side effects were recorded. RESULTS: Pain scores (0-10) were higher in the PCA group (on movement, day 1/day 2: 7.0/6.5) than in the EPI group (5.0/5.0) and the PSC group (4.0/3.5). Postoperative opioid consumption over 48 h was higher in the PCA group (51 mg) than in the EPI group (0 mg) and the PSC group (0 mg). There were no differences in functional recovery. Pruritus occurred more frequently in the PCA and EPI groups than in the PSC group. Patients receiving a PSC and EPI were more satisfied than those treated with PCA. INTERPRETATION: Analgesia with PSC catheters or EPI catheter is superior to PCA regarding pain levels, analgesic requirements, and patient satisfaction. There was no difference in functional outcome between the 3 groups. PMID- 17464607 TI - Bone regeneration 6 years after impaction bone grafting: a PET analysis. AB - BACKGROUND: Impacted morselized bone allograft in revision total hip arthroplasty for prosthetic loosening has gained widespread clinical use during the last decades. The clinical results are good but little is known about the bone regeneration in the graft. PATIENTS AND METHODS: 5 patients were revised with impaction of morselized frozen allograft and a cemented total hip arthroplasty (THA) due to loosening and osteolysis of a primary THA. We used positron emission tomography ([18F]-fluoride PET) to produce quantitative images of new bone formation in the allograft surrounding the femur stem 6 years after surgery. RESULTS: The 5 patients had previously been analyzed by [18F]-fluoride PET during the first year after surgery (Sorensen et al. 2003). During the first year, bone formation proceeded through the graft layer and reached the cement layer surrounding the femoral stems. The clinical and radiographic results were excellent at 6 years. PET analyses at 6 years showed that the bone metabolism was significantly reduced in most areas of the proximal femur, compared to the elevated activity during the first year after surgery, and also normalized compared to the contralateral healthy femur. Graft bone metabolism distal to the stem tip remained slightly increased. Small patchy areas of increased uptake remained along the proximal femoral stem, probably reflecting small volumes of fibrous healing. INTERPRETATION: The metabolism of the allografted bone had normalized compared to native bone, indicating full regeneration throughout the graft--and a good long-term prognosis for implant fixation. PMID- 17464608 TI - Charnley low-frictional torque arthroplasty in young rheumatoid and juvenile rheumatoid arthritis: 292 hips followed for an average of 15 years. AB - INTRODUCTION: In the early 1960s, Charnley was cautious with his patient selection for total hip replacement. As follow-up increased and confidence in the operation grew, younger patients were selected. We present our results of the Charnley LFA in young patients with rheumatoid and juvenile rheumatoid arthritis with a followup of up to 36 years. PATIENTS: We studied 292 Charnley low-friction arthroplasties in 195 young patients with an established diagnosis of rheumatoid arthritis. Their mean age at operation was 38 (12-50) years; 168 (58%) were receiving steroids and 79 (27%) were on non-steroidal antiinflammatory medication. The mean follow-up for the whole group was 15 (1-36) years. 24 patients could not be traced (33 hips), and 61 patients died (88 hips). RESULTS: 25 patients (41 hips) had had a revision. The main indication for revision was cup loosening. In the 85 patients (130 hips) attending follow-up, their mean age at surgery was 36 (17-50) years and the mean follow-up was 20 (10-36) years. 98% were pain-free or had no more than occasional discomfort, 44% claimed to have normal or near-normal function, while 62% had full or almost full range of movement of the replaced hip. Radiographically, 29 cups (22%) were considered to be loose. 1 stem (1%) was definitely loose and 2 stems (2%) were probably loose. With revision for any indication as the endpoint, the survival was 74% at 25 years follow-up. INTERPRETATION: The Charnley LFA continues to be an excellent hip replacement, even for very young rheumatoid arthritis patients. Wear and aseptic cup loosening are the main long-term problems. PMID- 17464609 TI - Neo-capsule tissue reactions in metal-on-metal hip arthroplasty. AB - BACKGROUND: Modern-generation metal-on-metal articulations have a high wear resistance and may therefore be able to improve the long-term performance of artificial joints. However, the biological effects are still under debate. This study was undertaken to review the histopathological changes in neo-capsule tissues of different metal-on-metal hip arthroplasties. METHODS: Neo-capsule tissue samples from 46 hips with modern second-generation metal-on-metal articulations (39 hip resurfacings and 7 non-cemented total hip replacements) with a variety of failure mechanisms were examined histopathologically and immunohistochemically. RESULTS: A distinct lymphocytic infiltration was found in all cases with in situ times of more than 7 months, consisting of CD20-positive B lymphocytes and CD3-positive T-lymphocytes and sometimes thinly distributed CD138 positive plasma cells without dominant T-cell or plasma cell infiltrates. INTERPRETATION: This distinct lymphocytic infiltration has not been reported in tissue analyses of metal- or ceramic-on-polyethylene hip replacements, and may therefore be considered to be a characteristic histological pattern of tissue reactions on metal particles and/or ions around metal-on-metal bearings. PMID- 17464610 TI - Tartrate-resistant acid phosphatase 5b (TRAP 5b) as a marker of osteoclast activity in the early phase after cementless total hip replacement. AB - BACKGROUND: After total hip replacement, increased bone metabolism is seen. A local periprosthetic osteopenia can be measured by dual-energy X-ray absorptiometry (DXA), but it is still unkown whether biochemical markers can be used to monitor the local remodeling at an earlier stage. PATIENTS AND METHODS: In this prospective study we compared the biochemical markers tartrate-resistant acid phosphatase 5b (TRAP 5b), bone ALP, osteocalcin and CrossLaps with periprosthetic DXA in 17 consecutive patients after uncemented total hip replacement. RESULTS: We found a highly significant early increase in TRAP 5b after 2 weeks and 6 weeks, which was followed by a densitometrically detectable decrease in bone mineral density after 26 weeks, especially in periprosthetic section Gruen zone 7. Bone ALP and osteocalcin levels as markers of osteoblast activity, and also Cross-Laps as a further marker of osteoclast activity, did not appear to allow any significant prediction of local bone remodeling. DISCUSSION: Our findings show that TRAP 5b is a sensitive parameter for monitoring of osteoclast activity after cementless total hip replacement, and may predict local osteopenia. PMID- 17464611 TI - Frozen sections of samples taken intraoperatively for diagnosis of infection in revision hip surgery. AB - BACKGROUND: The diagnosis of a suspected infected prosthesis is often difficult, but is important for the choice of treatment. Even at surgery, it is not easy to assess whether the prosthesis is infected or not--even though this may be important for the choice of surgical procedure. PATIENTS AND METHODS: We assessed the sensitivity, specificity, predictive value, and reliability of the results of the analysis of frozen sections from samples of tissues taken during revision hip surgery of 136 probably infected prostheses. Samples of tissues were taken to be analyzed immediately from frozen sections, to be processed on a routine basis later, and to be referred for bacteriological cultures. A finding of 5 or more polymorphonuclear leukocytes per field at a magnification of 400x was considered positive for infection. RESULTS: The analysis of frozen sections for infection was in agreement with the results of routine histopathology in 134 of 136 cases. Comparison with the results of culture showed a sensitivity of 85%, a specificity of 87%, a PPV of 79%, an NPV of 91%, and a Kendall's tau correlation coefficient of 0.72. INTERPRETATION: We believe that the method we have tested is of value in revision surgery when infection cannot be ruled out. PMID- 17464612 TI - Outcome after treatment of complications of Gamma nailing: a prospective study of 554 trochanteric fractures. AB - BACKGROUND: A high reoperation rate has been the main reason why Gamma nailing should not be recommended for routine use in the treatment of trochanteric fractures. We compared the outcome after reoperation to the outcome after primary surgery with Gamma nailing. METHODS: In a series of 554 patients, we compared the outcome in 52 patients who were reoperated with that in 502 patients who had no reoperations. We assessed mortality, pain, walking ability and habitat at follow up. RESULTS: The most common reason for reoperation was new fracture around the implant (17), local pain after healed fracture (11), nonunion (9) and cut-out (8). A second reoperation was required in 9/52 patients. The mortality was significantly lower in the reoperated cases at 30 days and at 1-5 years, but not at 120 days, and there were no significant differences in the other outcome parameters. INTERPRETATION: Reoperation did not lead to a worse clinical outcome, nor to increased mortality. PMID- 17464613 TI - Long-term results after closed reduction of latedetected hip dislocation: 60 patients followed up to skeletal maturity. AB - BACKGROUND: This retrospective study was undertaken because there is limited knowledge about the long-term results after closed reduction of late-detected hip dislocation. The aims were to evaluate the outcome after skeletal maturity and to find predictive factors for good and poor results. PATIENTS AND METHODS: The material included 60 patients (78 hips, 53 girls) treated during the period 1958 62. The primary treatment was skin traction for 36 (16-76) days. Closed reduction was performed in all hips except 4 that needed open reduction. The mean age at reduction was 20 (4-65) months. Hip spica plaster was worn for 9 (6-20) months. Within 3 years of the start of treatment, derotation femoral osteotomy was performed because of increased femoral anteversion in 35 patients. Later, 28 patients underwent additional surgery on the femur or acetabulum to improve femoral head coverage. Radiographs at the time of diagnosis and during follow-up to skeletal maturity were assessed. The average age of the patients at the most recent follow-up was 26 (15-42) years. RESULTS: The femoral head coverage normalized during the primary treatment and then decreased somewhat during the remaining growth period. The dysplasia of the acetabulum improved markedly during the first year after reduction. It continued to improve, but to a much lesser degree, until 8-10 years of age. A satisfactory radiographic outcome at skeletal maturity (Severin grades I and II) was obtained in 63% of the hips. Early derotation osteotomy of the femur did not improve the outcome. Avascular necrosis of the femoral head occurred in 14% of the hips. Risk factors for unsatisfactory outcome at skeletal maturity were high initial dislocation, steep acetabulum 1 year after reduction, reduced femoral head coverage at age 8-10 years, and avascular necrosis. INTERPRETATION: The specific risk factors and the radiographic outcome--with satisfactory long-term results in nearly two-thirds of the patients--would be valuable for comparison with outcome studies after more modern treatment regimes. PMID- 17464614 TI - Documentation of fracture severity with the AO classification of pediatric long bone fractures. AB - BACKGROUND: The AO comprehensive pediatric longbone fracture classification system describes the localization and morphology of fractures, and considers severity in 3 categories: (1) simple, (2) wedge, and (3) complex. We evaluated the reliability and accuracy of surgeons in using this rating system. MATERIAL AND METHODS: In a first validation phase, 5 experienced pediatric (orthopedic) surgeons reviewed radiographs of 267 prospectively collected pediatric fractures (agreement study A). In a second study (B), 70 surgeons of various levels of experience in 15 clinics classified 275 fractures via internet. Simple fractures comprised about 90%, 99% and 100% of diaphyseal (D), metaphyseal (M), and epiphyseal (E) fractures, respectively. RESULTS: Kappa coefficients for severity coding in D fractures were 0.82 and 0.51 in studies A and B, respectively. The median accuracy of surgeons in classifying simple fractures was above 97% in both studies but was lower, 85% (46-100), for wedge or complex D fractures. INTERPRETATION: While reliability and accuracy estimates were satisfactory as a whole, the ratings of some individual surgeons were inadequate. Our findings suggest that the classification of fracture severity in children should be done in only two categories that distinguish between simple and wedge/complex fractures. PMID- 17464615 TI - Injection of the subacromial-subdeltoid bursa: blind or ultrasound-guided? AB - BACKGROUND: Blind injection of the subacromial-sub-deltoid bursa (SSB) for diagnostic purposes (Neer test) or therapeutic purposes (corticosteroid therapy) is frequently used. Poor response to previous blind injection or side effects may be due to a misplaced injection. It is assumed that ultrasound (US)-guided injections are more accurate than blind injections. In a randomized study, we compared the accuracy of blind injection to that of US-guided injection into the SSB. PATIENTS AND METHODS: 20 consecutive patients with impingement syndrome of the shoulder were randomized for blind or US-guided injection in the SSB. Injection was performed either by an experienced orthopedic surgeon or by an experienced musculoskeletal radiologist. A mixture of 1 m'L methylprednisolone acetate, 4 mL prilocaine hydrochloride and 0.02 mL (0.01 mmol) Gadolinium DTPA was injected. Immediately after injection, a 3D-gradient T1-weighted magnetic resonance scan of the shoulder was performed. The location of the injected fluid was independently assessed by 2 radiologists who were blinded as to the injection technique used. RESULTS: The accuracy of blind and US-guided injection was the same. The fluid was injected into the bursa in all cases. INTERPRETATION: Blind injection into the SSB is as reliable as US-guided injection and could therefore be used in daily routine. US-guided injections may offer a useful alternative in difficult cases, such as with changed anatomy postoperatively or when there is no effective clinical outcome. PMID- 17464616 TI - Survivorship of the KUDO total elbow prosthesis--comparative study of cemented and uncemented ulnar components: 89 cases followed for an average of 6 years. AB - BACKGROUND: The Kudo total elbow prosthesis (TEP) is a well-established implant with good mid-term results. The ulnar component can be placed with or without cement, and the humeral component is normally placed without cement. METHODS: 89 Kudo type-5 total elbow prostheses were evaluated after a mean follow-up of 6 (1.7-11) years. The indication for joint replacement was rheumatoid arthritis in all cases. 49 prostheses were placed without cement. In 40 cases, the ulnar component was cemented and the humeral component was uncemented. RESULTS: In the uncemented group, 7 revisions had taken place. 3 of these ulnar components were shortstemmed and 4 were long-stemmed. No revisions had been performed in the hybrid group. In the uncemented group another 7 patients showed progressive radiolucencies, while 3 patients in the hybrid group showed progressive radiolucencies. INTERPRETATION: In this group of RA patients, the survivorship of the cemented ulnar component was better than that of the uncemented ulnar component. PMID- 17464617 TI - Failure mechanisms in uncemented Kudo type 5 elbow prosthesis in patients with rheumatoid arthritis: 7 of 49 ulnar components revised because of loosening after 2-10 years. AB - BACKGROUND: Both components of the Kudo type 5 elbow prosthesis can be inserted with or without the use of cement. There have been no reports on the use of this prosthesis with all components uncemented in patients with rheumatoid arthritis. PATIENTS AND METHODS: We reviewed 49 primary uncemented Kudo type 5 elbow prostheses, inserted in 36 patients with rheumatoid arthritis, after mean 6 (2 10) years. Patients were assessed clinically both pre- and postoperatively (pain, instability, motion, ulnar neuropathy) and radiographically. Furthermore, at the time of follow-up clinical outcome was assessed using the Elbow Function Assessment Scale. RESULTS: At review, 7 of 49 elbows had undergone revision because of symptomatic loosening of the ulnar component. In 42 unrevised elbows, clinical outcome was excellent in 29, good in 7, fair in 5, and poor in one. 31 of 42 elbows had no pain; 11 were painful at rest (VAS 1-2) and/or as a result of activity (VAS 1-8). With revision as endpoint, survival was 86% at 6 years. Intraoperative malpositioning of the ulnar component with a valgus or varus alignment of < 5 degrees was associated with worse survival. INTERPRETATION: We found an unexpectedly high rate of loosening of the ulnar component, which was associated with intraoperative malpositioning of the prosthesis. The ulnar component of this prosthesis should not be inserted without cement in patients with rheumatoid arthritis. PMID- 17464618 TI - Are distal radius fractures due to fragility or to falls? A consecutive case control study of bone mineral density, tendency to fall, risk factors for osteoporosis, and health-related quality of life. AB - BACKGROUND: A fracture of the distal radius is considered to indicate an increased risk of future fractures, especially a hip fracture. The main causes may be osteoporosis or a tendency to fall, separately or in combination. METHODS: 93 women and 5 men with a recent radius fracture and the same number of controls were measured with a heel-DXL and asked to complete one questionnaire on their quality of life (SF-36), and one on risk factors. RESULTS: The mean T-score of the patients was -2.1, and for the controls it was -1.9 (p = 0.3). The patients aged over 64 years had a history of falling more often than the corresponding controls (p = 0.01), but there was no difference in T-score. By contrast, patients 45-64 years of age showed a non-significant, lower T-score (p = 0.09), but there was no difference concerning their history of falling. For all other risk factors, no differences were found between the patients and the controls. There were significant differences between the patients and the controls in some of the functions in the SF-36, due to the radius fracture. INTERPRETATION: This study indicates that the underlying cause of a distal radius fracture may be different in patients aged 45-64 years and those who are more than 64 years old. PMID- 17464619 TI - Reviewer agreement in scoring 419 abstracts for scientific orthopedics meetings. AB - BACKGROUND: The selection of presentations at orthopedic meetings is an important process. If the peer reviewers do not consistently agree on the quality score, the review process is arbitrary and open to bias. The aim of this study was: (1) to describe the inter-reviewer agreement of a previously designed scoring scheme to rate abstracts submitted for presentation at meetings arranged by the Dutch Orthopedic Association; (2) to test whether the quality of reporting of submitted abstracts increased in the years after the introduction of the scoring scheme; and (3) to examine whether a review process with a larger workload had lower interrater agreement. METHODS: We calculated intraclass correlation coefficients (ICC) to measure the level of agreement among reviewers using the International Society of the Knee (ISK) quality-of-reporting system for abstracts. Acceptance rate and quality of the abstracts are described. RESULTS: Of 419 abstracts, 229 (55%) were accepted. Inter-reviewer agreement to rate abstracts was substantial (0.68; 95% CI: 0.47-0.83) to almost perfect (0.95; 95% CI: 0.92-0.97) and did not change over the eligible time period. A smaller proportion of abstracts were accepted after 2004. The mean ISK abstract score (with a maximum of 100 points) for accepted abstracts ranged from 60 (95% CI: 58-63) to 64 (95% CI: 62-66). The mean ISK abstract score for rejected abstracts varied from 46 (95% CI: 40-51) to 51 (95% CI: 47-55). Average scores for accepted and rejected abstracts did not change with time. The degree of workload of the reviewers did not influence their level of agreement. INTERPRETATION: The ISK abstract rating system has an excellent interobserver agreement. Other scientific orthopedic meetings should consider adopting this ISK rating system for further evaluation in a local or international setting. PMID- 17464620 TI - The effect of recombinant human bone morphogenetic protein-2 on the osteogenic potential of rat mesenchymal stem cells after several passages. AB - BACKGROUND: Since the osteogenic potential of bone marrow derived mesenchymal stem cells (BMSCs) becomes reduced with passage, establishment of culture condition that permit the rapid expansion of BMSCs while retaining their potential for differentiation is needed for clinical application. Bone morphogenetic proteins stimulate osteogenic differentiation in mesenchymal progenitor cells as well as increase stem cell numbers. Thus, we analyzed the effect of recombinant human bone morphogenetic protein-2 (rhBMP-2) on the osteogenic potential of rat BMSCs over several passages. MATERIAL AND METHODS: Osteogenic differentiation in vitro was evaluated in terms of the alkaline phosphatase (ALP) activity and the osteocalcin (OC) concentration in the supernatants, and the expression of ALP and OC mRNA in the cultured cells. For in vivo osteogenesis, BMSCs cultured with and without rhBMP-2 through all passages were implanted into athymic mice. RESULTS: The levels of osteogenic markers were significantly higher in the cells of the BMP(+) group than in the cells of the BMP(-) group, although they decreased with passage irrespective of whether or not rhBMP-2 was added. Similar to the in-vitro experiments, there was a greater degree of bone and cartilage tissue formation in the BMP(+) group over all passages. INTERPRETATION: From our results, osteogenic potential can be maintained even in BMSCs that have been passaged several times in the presence of rhBMP-2. These cells are capable of inducing and participating in bone formation and can be used for clinical applications. PMID- 17464621 TI - Compartment syndrome of the calf following total knee arthroplasty--a case report of a highly unusual complication. PMID- 17464622 TI - Necrotizing fasciitis caused by TSST-1 producing penicillin-sensitive Staphylococcus aureus--a case report. PMID- 17464624 TI - Releasing psychiatry from the constraints of categorical diagnosis. AB - OBJECTIVE: The aim of this study was to make a case for freeing psychiatry from the limits of categorical diagnosis. CONCLUSIONS: From Pinel through to Kraepelin, Bleuler, DSM-IV and ICD-10, clinical and research activities in psychiatry have all depended upon categorical classification. Recent decades have seen increasing argument in favour of dimensional measurement, description and prediction, in part because invalidities in categorical classification are becoming impossible to deny. Some suggestions for developing useful dimensional approaches are made. PMID- 17464626 TI - Dysphoric about dysphoria: towards a greater conceptual clarity of the term. AB - OBJECTIVE: The aim of this study was to conceptualize dysphoria in a way that is clearer, better delineated and more clinically useful, after reviewing its meanings in a variety of psychopathological contexts and the consequent conceptual confusion. CONCLUSIONS: The current semantic status of dysphoria is most unsatisfactory. Its definitions are usually too broad or too simplistic and, therefore, not clinically useful. There is no agreement on what the term means. In this article, dysphoria is conceptualized primarily as a complex emotional state, consisting of intense unhappiness/discontent and irritability. In addition, dysphoria is often, though not invariably, accompanied by certain cognitive and behavioural features. This article reviews relationships between dysphoria and related emotional states and various psychiatric disorders, and proposes differentiations that have implications for clinical practice. The role of dysphoria in mixed states within bipolar disorder is particularly in need of further study. PMID- 17464627 TI - Implementation of a first presentation psychosis clinical pathway in an area mental health service: the trials of a continuing quality improvement process. AB - OBJECTIVES: The aim of this study was to describe the process of improving early psychosis services in a generic Victorian adult area mental health service from 1998 to 2004 and to give consideration to whether to implement a first presentation psychosis clinical pathway and how that should be done. METHOD: A description will be given of the quality improvement process and practical difficulties in the sequential implementation of guidelines then pathways for early psychosis. RESULTS: We experienced many of the commonly reported difficulties in quality improvement and implementation of evidence-based practice in real clinical settings. CONCLUSIONS: Developing and implementing an evidence based first presentation psychosis clinical pathway was a difficult but rewarding learning experience. PMID- 17464628 TI - Predicting treatment response during the acute phase of hospitalization. AB - OBJECTIVE: This study sought to determine if an initial assessment for an acute inpatient population was able to predict treatment response as measured by progress through clinical pathways. In addition, we sought to analyze the impact of comorbidity on treatment response. METHOD: A total of 170 adults (18-75 years) admitted for an acute inpatient hospitalization, received a full assessment including the Brief Symptom Inventory. Based on assessment results, patients were placed on one of five clinical pathways. Treatment response was measured by the number of days spent on each level of the clinical pathway. RESULTS: Results revealed an unexpected, significant negative correlation between symptom severity on admission and movement through the clinical pathway; that is, patients with more severe symptomatic presentations at assessment made more rapid progress through the clinical pathways. However, comorbidity showed the strongest relationship with treatment response. Taken together, these variables explained 52% of the variance in treatment response. CONCLUSIONS: Responsiveness to antipsychotic medications, as well as high levels of subjective distress and the consequent motivation to achieve relief, may help explain the unexpected relationship between symptom severity and treatment response. Findings also demonstrated the utility of an initial assessment in predicting treatment response. PMID- 17464629 TI - The limitations of advance directives and statements in mental health. AB - OBJECTIVE: Advance directives and statements in mental health are defined and the necessary conditions for making them explained. The practical limitations on their usefulness are discussed. CONCLUSION: Legal recognition of advance directives and statements is limited. There has been little research into their use and impact on care and what little exists, is equivocal in results. PMID- 17464630 TI - A survey of combination antidepressant use in Australia. AB - OBJECTIVE: The aim of this study was to survey doctors working in psychiatry in Australia about the practice of using two antidepressants simultaneously. METHOD: A postal survey was sent to all doctors in psychiatry in Australia enquiring about their prescribing history and their attitudes to combination antidepressants and related issues. RESULTS: Seventy-nine percent of respondents had used combination antidepressants. The most frequently reported combination was a selective serotonin reuptake inhibitor combined with a tricyclic antidepressant. Combinations of mirtazepine with venlafaxine and other antidepressants were the next most frequently used. Seventeen percent of respondents reported having seen a complication from combination antidepressants, 75% believed that Australian GPs should be given information on the use of combination antidepressants, 89% wished for more information on this topic, and 88% believed patients had a right to be informed of this option in their treatment. Use of combination antidepressants was more frequent than exceeding the recommended maximum dose of an individual antidepressant. CONCLUSION: Combination antidepressants are used far more frequently in Australia than suspected previously. Research into safe and evidence-based practice is strongly indicated. PMID- 17464631 TI - Bipolar disorder: from psychoeducational to existential group therapy. AB - OBJECTIVE: The aim of this study was to assess the importance of addressing psychoeducational and existential issues in group therapy for patients with bipolar disorder. METHOD: The group consisted of 10 bipolar patients. Group sessions were held for 2 hours weekly for 3 months and then monthly up to 2 years. Group topics focused on psychoeducation, cognition and existential issues. RESULTS: Over 2 years, two members had a minor relapse that did not require hospitalization. Adherence with pharmacotherapy and follow up was 100%. All members reported significant improvements in family and social function and nine members remained employed in full-time jobs. There was a high level of satisfaction with the group process. CONCLUSION: Addressing existential and psychosocial issues in bipolar disorder group therapy may improve outcome. This approach deserves further thorough investigation. PMID- 17464632 TI - The Green Card Clinic: overview of a brief patient-centred intervention following deliberate self-harm. AB - OBJECTIVES: The aim of this study was to present an overview of the Green Card Clinic, a novel brief intervention service for patients presenting to the emergency department following deliberate self-harm (DSH) or with suicidal ideation, to examine its effectiveness in terms of service utilization, and patient and clinician feedback, and to explore the correlates of repeated DSH. METHOD: The aims and structure of the Green Card Clinic are described. We highlight our patient-centred approach involving self-identification of difficulties from a list of problem areas, coupled with tailored intervention strategies. Relevant data are presented and characteristics of repeat DSH patients are compared to the first-episode group. RESULTS: Between 1998 and 2005, 456 DSH patients were referred to the clinic. Of these, 75% (n = 344) attended the first session, 43% (n = 197) the second session, 26% (n = 117) the third session, and 16% (n = 73) completed a 3-15 month follow-up. Clinic attenders (mean age 31.6 years, 57% female) reported a diverse range of self-identified problems and repeat DSH patients reported worse depression, poorer health-related behaviours, and a greater number of problems than those presenting after first episode DSH. CONCLUSIONS: The clinic achieved high rates of first session attendance. This may have been attributable to the use of a few specific strategies aimed at increasing compliance, such as the green card, next-day appointments and assertive follow-up of non-attenders. For repeat self-harmers, we advocate an approach aimed at 'lifestyle change' rather than based on current psychological stressors. The Green Card Clinic service, involving a range of interventions tailored to meet the multitude of presenting needs, appears to be an acceptable and flexible approach to brief intervention for DSH. PMID- 17464633 TI - An institute with walls. PMID- 17464634 TI - Hallucinations in adolescents with post-traumatic stress disorder and psychotic disorder. AB - OBJECTIVE: The aim of the study was to compare the phenomenology of hallucinations in hospitalized adolescents diagnosed with post-traumatic stress disorder (PTSD) and psychotic disorder. METHODS: Diagnoses were ascertained in 66 adolescents consecutively admitted to an acute adolescent unit in Australia. Hallucinations were examined to compare their phenomenology in PTSD with psychotic disorder. RESULTS: On most measures, hallucinations did not differ significantly in form or content between those adolescents with PTSD and those with psychotic disorder. In a minority of patients with PTSD, the hallucinations reflected a previous traumatic experience. CONCLUSIONS: Hallucinations were found to be unreliable in distinguishing adolescent inpatients with PTSD from those with psychotic disorder. PMID- 17464635 TI - Consultant caseload management. AB - OBJECTIVE: This study was designed to survey a different model of working as a consultant in a child and adolescent mental health service to address the problem of a long waiting list. METHODS: A log of referrals was kept from date of entry and when patient were seen. RESULTS: The main outcome was in reducing the waiting time from 22 weeks to 8.4 weeks. CONCLUSIONS: The clinical implication is for consultants to consider using a consultation model in child and adolescent mental health services. PMID- 17464636 TI - An integrated approach to general practice mental health training: the importance of context. AB - OBJECTIVE: The high prevalence of mental disorders and the barriers to detection and treatment of these in general practice are well recognized. As such, the government has placed great emphasis on training general practitioners (GPs) in primary care mental health and on the provision of support for GPs in the delivery of such services. The current paper aims to evaluate a local, rural training program in mental health for GPs. We hypothesized that local 'context driven' training would lead to increased knowledge and reported change in practice by GPs with mental health patients. METHOD: Locally developed and delivered 'Level 1' training was offered to GPs through the Better Outcomes in Mental Health Care initiative. The training was provided with 6-hour workshops covering mental health assessment and management planning. The training was evaluated through pre- and 6-week post-questionnaires assessing attitudes and practice with respect to treatment of patients with mental health problems. RESULTS: Forty-nine GPs from the Loddon Campaspe Southern Mallee region of Victoria participated in the training. Following the 6-hour workshop, there was an increase in reported use of psychoeducation for patients with depression, use of cognitive behavioural therapy for patients with anxiety, and ease in obtaining advice to assist with the management of psychosis. CONCLUSIONS: GP mental health education should take into consideration the local context, cover systems issues as well as skills development, and aim to develop personal relationships between mental health clinicians and GPs to enhance outcomes. PMID- 17464637 TI - The Top End Mental Health Services General Practice Clinic: an initiative for patients with serious mental illness. AB - OBJECTIVE: The aim of the present study was to provide an overview of the benefits of establishing a general practice clinic within a community mental health service. The clinic was established via collaboration between the Division of General Practice and the Mental Health Service. METHODS: Data collected over 19 months during the operation of the General Practice Clinic were compared to national benchmarks for similar general practice data. RESULTS: The clinic was able to offer much longer appointments than would normally be available in most general practices. This allowed for comprehensive screening of serious illness as well as health promotion activities. CONCLUSIONS: The General Practice Clinic was able to address the significant health needs for patients with serious mental illness in a manner that was ahead of national benchmarks for this patient group. Barriers to care in the patient group, such as stigma and non-attendance, were overcome by coordination between the general practitioners staffing the clinic and their collaboration with the case managers and reception staff of the service. PMID- 17464638 TI - The interface between religion and psychosis. AB - OBJECTIVE: This paper aims to explore the interface between religion and psychosis, and to comment on its relevance in clinical practice. METHOD: The context of religious psychotic phenomena is briefly discussed, leading to an examination of the biological substrates of religious experiences, the hypothesized process of religious psychotic symptom formation, and the clinical implications when assessing religious delusions. A PubMED search was conducted to identify original research and review articles of relevance to the discussion. RESULTS: Religion is an enduring theme in psychosis, the understanding of which can be assisted by distinguishing between religion as a culture and religiosity as pathology. There are strong arguments for the involvement of temporolimbic instability in the generation of religious psychotic symptoms. CONCLUSIONS: Psychosis can be conceptualized as the manifestation of aberrant perceptual and/or integrative processes. The prevalence of religion as a psychotic theme may be explained by its central cultural role, the implication of temporolimbic overactivity in the pathogenesis of some cases of psychosis, and the tendency to interpret intense or discrepant perceptual events as spiritual. In the clinical setting, the determination of religious delusions can be challenging at times. In addition to seeking advice on unfamiliar religions, a thorough assessment of the dimensions of religious beliefs and symptoms of neurocognitive dysfunction can be useful. PMID- 17464639 TI - A tale of two cities: a neuroimaging investigation of Melbourne-Sydney rivalry comparing cortical thickness in healthy adults. AB - OBJECTIVE: We sought to identify neurobiological correlates of Melbourne-Sydney rivalry through neuroimaging measures of a key brain region involved in cognitive and emotional regulation. METHOD: Twenty subjects from each city were recruited from two large neuroimaging databases, and were scanned on a GE Signa 1.5 T magnetic resonance imaging scanner. Cortical thickness of the anterior cingulate cortex (ACC) was measured using a tessellated mesh method, after image segmentation. These measures were compared with key sporting, financial and academic variables. RESULTS: Residents of Melbourne had a significantly thicker ACC (p < 0.0001) than Sydney residents, and this difference remained significant when age and intracranial volume were controlled for (p = 0.001). This difference mirrored that in variables measuring wealth, sporting and academic success. CONCLUSIONS: The thinner ACC seen in Sydney-siders may reflect the effects of increased stress due to elevated property prices, relative lack of sporting success and other variables. An alternative explanation is that a thinner ACC is the result of increasing cortical refinement and efficiency, and a marker of a more mature city. However, if these findings are a result of latitudinal effects, this may have significant implications for residents of more northern regions of the Australian continent. PMID- 17464640 TI - A possible behavioural and cognitive phenotype for the 47,XYY karyotype in a pre pubertal child. AB - OBJECTIVE: The aim of this study was to describe the case of a 9-year-old boy with 47,XYY genotype and possible behavioural phenotype involving 'internalizing' and 'externalizing' symptoms and cognitive phenotype of deficient spatial memory. CONCLUSIONS: Longitudinal study of 47,XYY cases recognized prenatally are needed to confirm the preliminary findings described in this report. PMID- 17464642 TI - Is there a systematic bias in the prosecution of psychiatric patients who assault staff? PMID- 17464649 TI - The democratisation of psychiatry. AB - OBJECTIVE: To explore the decline in expertise of the consultant psychiatrist and its replacement by a range of professional colleagues. CONCLUSION: Averting a devaluing of expertise in Australasia requires thoughtfulness, willingness to quietly and confidently contribute, and professional behaviour. There are early promising developments. PMID- 17464650 TI - Is deep brain stimulation a form of psychosurgery? AB - OBJECTIVE: To examine the potential for the experimental treatment of deep brain stimulation for neuropsychiatric disorders, and to debate the argument that it should be considered another form of psychosurgery. CONCLUSIONS: Psychosurgery is an old term with considerable pejorative connotations. It should be replaced with the more descriptive and accurate 'neurosurgery for psychiatric disorders'. Moreover, neurosurgery should reflect ablative neurosurgery, and surgery for brain stimulation should be categorised as brain stimulation rather than neurosurgery, or indeed psychosurgery. This will prevent legislative restrictions on the development of brain stimulation techniques and not tar them with the lobotomy brush. PMID- 17464651 TI - Clickety-click: e-mental health train on track. AB - OBJECTIVE: Experts forecast that the Internet will bring about radical change in healthcare. This paper aims to summarise evidence concerning the efficacy of Internet-based programs in the delivery of mental health care and its implications for mental health professionals. It also describes those Internet programs which are both of demonstrated effectiveness in the treatment of anxiety or depression and available without restriction or cost to the public. CONCLUSIONS: There is evidence that Internet-based programs can improve a range of mental health conditions. Some of these programs could be used by psychiatrists and other mental health professionals as an adjunct to clinician based treatment. However, clinicians need to know which programs work and how they can be accessed. PMID- 17464652 TI - We can't find the solution until we know the problem: understanding the mental health nursing labour force. AB - OBJECTIVE: Difficulties recruiting and retaining adequate numbers of mental health nurses have been extensively documented in the Australian literature. The continued increase in the average age of practicing mental health nurses has intensified concerns that a workforce crisis is rapidly approaching. Despite the urgency of this situation, there has been no comprehensive, co-ordinated collection of labour force data. The aim of this paper is to synthesise and present labour force data gathered from various official sources to more clearly identify and articulate the nature and extent of the problem. METHOD: Relevant labour force data was obtained from reports produced by the Australian Institute of Health and Welfare and the Victorian Department of Human Services. Information was collated, synthesised and, in some cases, re-analysed to provide a clearer picture of the current national and Victorian mental health nursing labour force, as well as requirement and supply projections. RESULTS: Findings are consistent with conclusions in the available literature but suggest that the magnitude of the problem is likely to be greater than previously anticipated. CONCLUSIONS: The systematic and coordinated collection of mental health nursing labour force data is crucial in order that appropriate interventions can be implemented and evaluated. PMID- 17464653 TI - Contemporary management of aggression in an inner city mental health service. AB - OBJECTIVES: From 2002 to mid 2003, Royal Brisbane and Women's Hospital Mental Health experienced increased patient - staff aggression and staff injury which resulted in staff conflict and recruitment difficulties. Strategies introduced to reduce the frequency and impact of aggression in the mental health service were evaluated. METHOD: By mid 2003, incident data indicated increasing aggressive incidents. Based on this, an aggression management strategy was developed which included improved staff communication, new acute pharmacological treatment protocols, mandatory staff aggression management training, personal alarms and aggression risk assessment tools. RESULTS: Following the introduction of the strategy in early 2004, there was a reduction of 40% in aggressive incidents and a 56% reduction in staff injuries in 2005 compared to 2003 levels. A more assertive approach to tranquillisation was not associated with an increased adverse event rate for patients. CONCLUSIONS: A co-ordinated strategy can contain and reduce aggressive incidents in acute inpatient mental health settings. These strategies are transferable to other health settings including Emergency Departments. PMID- 17464654 TI - A recovery-based outreach program in rural Victoria. AB - OBJECTIVE: A recovery-based outreach program for people with severe mental illness in regional Victoria is described. The paper covers a description of the program, the services provided and outcomes achieved. The program emphasized active collaboration between patients and clinicians as outlined in the collaborative recovery model and recognized that recovery from mental illness is an individual, personal process. CONCLUSIONS: The program provided service to 108 people over 3 years and had a positive impact on clinicians, patients and carers. The benefits of recovery orientation, multidisciplinary teams, collaborative relationships and carer involvement are discussed. The paper highlights the need for a focus on recovery and comprehensive care for people with severe mental illness. PMID- 17464655 TI - Outcome in a specialist referral clinic for mood disorders: a qualitative and quantitative review. AB - OBJECTIVE: The aim of this study was to determine whether short-term outcome for those referred with a depressive disorder could be predicted from baseline clinical assessment and identify factors contributing to outcome in a clinic operating to a subtyping diagnostic and aetiologically weighted management model. METHOD: This pilot study involved a consecutive series of 85 patients referred to the Depression Clinic at the Black Dog Institute. RESULTS: Globally assessed baseline prediction was associated with short-term outcome. Comparably high rates of improvement were evident in those with bipolar, melancholic and non melancholic subtypes, and somewhat lower in those diagnosed with a 'secondary depression'. Qualitative and quantitative analyses established that outcome was most clearly associated with referral source, degree of take up of recommendations and implementation of psychotropic drug strategies. CONCLUSIONS: Study results encourage more definitive and comparative study designs to be derived. PMID- 17464656 TI - Perceptions of mental health service delivery among staff and Indigenous consumers: it's still about communication. AB - OBJECTIVE: A needs analysis was undertaken to determine the quality and effectiveness of mental health services to Indigenous consumers within a health district of Southern Queensland. The study focussed on identifying gaps in the service provision for Indigenous consumers. Tools and methodologies were developed to achieve this. METHOD: Data were collected through the distribution of questionnaires to the target populations: district health service staff and Indigenous consumers. Questionnaires were developed through consultation with the community and the Steering Committee in order to achieve culturally appropriate wording. Of prime importance was the adaptation of questionnaire language so it would be fully understood by Indigenous consumers. Both questionnaires were designed to provide a balanced perspective of current mental health service needs for Indigenous people within the mental health service. RESULTS: Results suggest that existing mental health services do not adequately meet the needs of Indigenous people. CONCLUSIONS: Recommendations arising from this study indicate a need for better communication and genuine partnerships between the mental health service and Indigenous people that reflect respect of cultural heritage and recognises the importance of including Indigenous people in the design and management of mental health services. Attention to the recommendations from this study will help ensure a culturally appropriate and effective mental health service for Indigenous consumers. PMID- 17464657 TI - Aboriginal child and adolescent mental health: a rural worker training model. AB - OBJECTIVE: The Third National Mental Health Plan places a strong emphasis on the development of an Aboriginal mental health workforce. This paper documents the establishment, implementation and initial evaluation of the Aboriginal and Torres Strait Islander Child and Adolescent Mental Health Traineeship Program, a partnership initiative involving Hunter New England Area Health Service (HNEAHS), Hunter New England Aboriginal Mental Health (HNEAMH) and the Department of Psychological Medicine at the Children's Hospital at Westmead (CHW), with guidance and input from additional collaborators. CONCLUSIONS: The program includes: (i) employment as a child and adolescent mental health worker and professional support and supervision through HNEAHS; (ii) a mentoring program provided through HNEAMH; (iii) formal academic studies in Aboriginal Mental Health; and (iv) a clinical education and supervision program conducted through the Department of Psychological Medicine, CHW. Initial feedback suggests that this is a promising program to train Aboriginal child and adolescent mental health workers. Further evaluation will provide information about its viability and effectiveness in providing an integrated, collaborative child and adolescent mental health service for Aboriginal and Torres Strait Islander children and their families. PMID- 17464658 TI - Six months in Aceh. AB - OBJECTIVE: Brief case histories are presented of people with schizophrenia treated in the Indonesian province of Aceh, where the author worked as a clinical psychologist for Medecines Sans Frontieres in 2005. Aceh was severely affected by the December 2004 tsunami, with significant destruction and loss of life. CONCLUSIONS: The case studies highlight the needs of patients and the current opportunities to establish more effective mental health services. The role of culture as a significant consideration and a possible barrier to accessing care is also discussed. PMID- 17464659 TI - Cochrane's problem: psychoanalysis and anejaculation. AB - OBJECTIVE: The stimulus inciting Archie Cochrane's interest in scientific medicine was his own ill health and the inability of psychoanalysis to cure his problem of sexual dysfunction. The aetiology and modern management of anejaculation will be considered. CONCLUSIONS: The treatment undertaken by Cochrane failed because his condition was organic. Evidence-based medicine was founded upon Cochrane's critical evaluation of his own psychoanalytic psychotherapy. PMID- 17464660 TI - Gunfire, humour and psychotherapy. AB - OBJECTIVE: To reflect on the humour associated with one's cultural heritage, the mechanisms of humour in the cultures of many, and its role in the processes of psychotherapy. CONCLUSIONS: The practice of psychotherapy is enhanced by the therapist who maintains a sense of humour, who appreciates its development in his/her patients, and who respectfully uses it to assist in building the therapeutic alliance. Therapy is not so serious that laughter need be excluded from it. PMID- 17464661 TI - Cannabinoid hyperemesis: marijuana puts patients in hot water. AB - OBJECTIVE: This paper describes a case of cannabinoid hyperemesis, a curious, emerging condition associated with long-term cannabis abuse. The authors wish to facilitate its recognition and management by clinicians. CONCLUSIONS: Cannabinoid hyperemesis is a cluster of symptoms characterized by chronic cannabis abuse, cyclical vomiting and compulsive bathing behaviours. The condition can be missed unless an adequate history is taken of illicit drug ingestion. It can be confused with obsessive-compulsive disorder. The treatment consists of supportive measures in the acute phase, followed by complete abstinence from cannabis. PMID- 17464664 TI - Hepatitis C and psychiatry. PMID- 17464675 TI - Systematic review of case management. AB - The purpose of the present paper was to review the current state of evidence for types of case management, focusing on the last 10 years since publication of the Cochrane Systematic Reviews of case management and assertive community treatment. A literature review of electronic databases from 1995 to the present to identify recent research on psychiatric case management, both original studies and reviews, was carried out. Original articles were organized on basis of year of study, experimental group and outcome variables to determine patterns. Sixty relevant papers were located. Thirty-nine are reports of experimental trials of types of case management and 21 are reviews or discussion papers. The focus of research is on assertive community treatment or intensive case management, with only five papers on other forms of less intense case management. Numerous outcomes have been examined, of those examined often enough to draw meaningful conclusions only one, engagement with services, has been consistently positive. All other outcomes have produced mixed results. The strength of findings in favour of case management has weakened over time. A heterogeneous group of experimental designs limits comparisons. Numerous issues with methodology and definitions of types of case management have beset research in this field. Assertive types of case management (including assertive community treatment and intensive case management) are more effective than standard case management in reducing total number of days spent in hospital, improving engagement, compliance, independent living and patient satisfaction. More important than the type of service configuration is to understand the clinical criteria of the services provided and their effectiveness. PMID- 17464676 TI - What do dopamine transporter and catechol-o-methyltransferase tell us about attention deficit-hyperactivity disorder? Pharmacogenomic implications. AB - The purpose of the present paper was to review studies of two candidate genes for attention deficit-hyperactivity disorder (ADHD) and to separate aetiological from therapeutic effects. Genomic studies of ADHD were reviewed for candidate dopamine genes and studies selected to distinguish catechol-o-methyltransferase (COMT) and dopamine transporter (DAT-1) effects. Pharmacogenomic findings for the COMT gene were in agreement with the 1977 observations of Sprague and Sleator, who reported that at low psychostimulant doses, children with ADHD showed a remarkable improvement on a short-term memory test at all levels of task load, whereas at higher doses, there was a significant decrement in performance on the more difficult versions of the task, corresponding to an 'inverted 'U' shaped curve'. Recent studies show that individuals with the homozygous COMT (valine/valine) genotype demonstrated improvement following psychostimulant treatment, because their tonic dopamine (DA) levels were low, whereas the homozygous COMT (methionine/methionine) individuals, who already have high initial prefrontal cortex (PFC) dopamine levels performed more poorly after medication, in tasks with high working memory load. On the other hand aetiological findings for DAT-1 gene were heterogenous, but more often positive than for COMT. Contrasting findings for COMT and DAT-1 may best be considered in terms of prediction of medication response in ADHD in the case of COMT, but in aetiological terms in the case of DAT-1, which is abundant in the striatum and possibly plays a greater role in determining hyperactivity and impulsivity, than working memory deficiencies. PMID- 17464677 TI - Empirical investigation of two assumptions in the diagnosis of DSM-IV major depressive episode. AB - OBJECTIVE: The aim of the current study was to examine two major assumptions behind the DSM-IV diagnosis of major depressive episode (MDE): that depression represents a distinct category defined by a valid symptom threshold, and that each depressive symptom contributes equally to the diagnosis. METHODS: Data were from the Australian National Survey of Mental Health and Wellbeing. Participants consisted of a random population-based sample of 10 641 community volunteers, representing a response rate of 78%. DSM-IV diagnoses of MDE and other mental disorders were obtained using the Composite International Diagnostic Interview, version 2.0. Analyses were carried out on the subsample of respondents who endorsed either depressed mood or loss of interest (n =2137). Multivariate linear regression analyses examined the relationship between the number and type of symptoms and four independent measures of impairment. RESULTS: The relationship between the number of depressive symptoms and the four measures of impairment was purely linear. Three individual symptoms (sleep problems, energy loss, and psychomotor disturbance) were all independent predictors of three of the four measures of impairment. CONCLUSIONS: Counting symptoms alone is limited in guiding a clear diagnostic threshold. The differential impact of individual symptoms on impairment suggests that impairment levels may be more accurately estimated by weighting the particular symptoms endorsed. PMID- 17464678 TI - Symptom profile of major depressive disorder in women with eating disorders. AB - OBJECTIVE: Based on the well-documented association between eating disorders (EDs) and affective disorders, the patterns of comorbidity of EDs and major depressive disorder (MDD) were investigated. The temporal relation between EDs and MDD onset was analyzed to determine differences in the course and nature of MDD when experienced prior to versus after the onset of the ED. METHOD: Lifetime MDD and depressive symptoms were assessed in 1371 women with a history of ED. The prevalence of MDD was first explored across ED subtypes, and ages of onset of MDD and EDs were compared. Depressive symptoms were examined in individuals who developed MDD before and after ED onset. RESULTS: The lifetime prevalence of MDD was 72.9%. Among those with lifetime MDD (n =963), 34.5% reported MDD onset before the onset of ED. Those who experienced MDD first reported greater psychomotor agitation (OR =1.53; 95%CI =1.14-2.06), and thoughts of own death (but not suicide attempts or ideation; OR =1.73; 95%CI =1.31-2.30). Among individuals who had MDD before ED, 26.5% had the MDD onset during the year before the onset of ED; 67% of individuals had the onset of both disorders within the same 3 year window. CONCLUSION: Clinicians treating individuals with new-onset ED or MDD should remain vigilant for the emergence of additional psychopathology, especially during the initial 3 year window following the onset of the first disorder. PMID- 17464679 TI - Judged effectiveness of differing antidepressant strategies by those with clinical depression. AB - OBJECTIVE: To obtain views of an Australian sample of individuals who had experienced a clinical depressive episode and examine the judged effectiveness of a wide range of professionally recommended, self-help and less orthodox antidepressant strategies. METHOD: A survey was posted on the Black Dog Institute website until a large sample had been derived. Inclusion criteria resulted in a sample of 2692 respondents. RESULTS: Effectiveness and benefit-burden scores (adjusting for not fully testing a strategy) were quantified for 31 strategies that had been trialled by at least 100 respondents. Many self-help strategies rated as strongly as professionally recommended strategies, with exercise being rated extremely highly. Intra-class comparisons of ratings identified some differentiation of antidepressant drugs but similar results for differing psychotherapies, while alternative drug strategies tended to return lower ratings than most formal antidepressant drugs. CONCLUSIONS: Study results, together with results from mental health literacy studies, advance our understanding about consumers' views about the frequency of trialling quite differing strategies and offer some information about their judged effectiveness. PMID- 17464680 TI - Have education and publicity about depression made a difference? Comparison of prevalence, service use and excess costs in South Australia: 1998 and 2004. AB - OBJECTIVE: To identify changes in depression, its management and associated excess costs, between 1998 and 2004 in South Australia. METHODS: A face-to-face Health Omnibus Survey was conducted in 2004 among 3015 randomly selected participants aged 15 years and over, who were a random and representative sample of the South Australian population, and this was compared with a survey conducted in 1998 that used the same methodology. The main outcome measures were prevalence of depression detected by the Mood Module of the Primary Care Evaluation of Mental Disorders (PRIME-MD); use of health services; health-related quality of life assessed by the Assessment of Quality of Life; estimates of excess costs and demographic data. RESULTS: There was no significant change in the overall prevalence of depression, although there was a significant decrease in respondents with other depressions, and a non-significant increase in those with major depression. No significant differences in the mean number of PRIME-MD depression symptoms were reported. Greater use of predominantly non-medical treatment services and antidepressants were reported by both those with depression and those without depression. There was a marked increase in the associated excess costs of depression. CONCLUSIONS: There has been no significant improvement in the prevalence of depression and its associated morbidity and financial burden in the South Australian community between 1998 and 2004, despite a number of professional and community education programmes. It is possible that without these efforts and the increased treatment reported on in this survey, there may have been an increase in the prevalence of depression and an even greater financial burden. However, it is also possible that community services for the provision of treatment for depression have not been able to implement research strategies that have been demonstrated to be effective. PMID- 17464681 TI - Neurocognitive function following remission in major depressive disorder: potential objective marker of response? AB - OBJECTIVE: Neurocognitive deficits have been widely reported in patients with mood disorders. However, relatively little is known of the short-term trajectory of neurocognitive improvement once treatment has been initiated. METHOD: A neurocognitive test battery was administered to unipolar depressed (major depressive disorder, MDD) patients (aged 18-65 years) who had been medication free for at least 6 weeks, and to healthy controls. Patients were then treated according to clinical need, predominantly with standard pharmacotherapy, and all participants were followed up within 6 months. RESULTS: Of the 25 MDD patients who returned at follow up, 11 were defined as remitted and 14 as not remitted. Significantly less baseline psychomotor dysfunction was observed in patients who remitted compared to those who did not (effect size, d =0.78, 95% confidence interval (CI) =0.07-1.44). Analysis of the change scores between assessments revealed a significantly greater improvement in verbal memory in patients who remitted compared to those who did not (d =0.73, 95%CI =0.03-1.39). CONCLUSIONS: This preliminary report suggests that there may be distinct temporal trajectories of neurocognitive improvement following remission in MDD. Aspects of neurocognitive functioning should be examined further as a means of providing a useful objective marker of treatment response. PMID- 17464682 TI - Effects of childhood experiences on cortisol levels in depressed adults. AB - OBJECTIVE: To evaluate the impact of childhood abuse and parental bonding on cortisol levels in depressed adults. METHODS: Mean afternoon cortisol levels were measured in 192 depressed adult patients at the beginning of a treatment trial. Childhood experiences of physical and sexual abuse were ascertained by interview, and perceived parenting by self-report. RESULTS: Maternal affectionless control, childhood sexual and physical abuse were all associated with cortisol levels. CONCLUSION: Childhood experiences, especially maternal affectionless control, appear to be related to hypothalamic pituitary adrenal axis function in depressed adults. PMID- 17464683 TI - Parental bonding and suicidality in adulthood. AB - OBJECTIVE: The short-term effect of an adverse parental child rearing style on suicidality in adolescence has been extensively discussed. Nevertheless, little is known about the long-term effect of adverse parental child rearing on lifetime suicidality in adulthood. So the present study aims to examine the relation between parental bonding on the one hand and suicidality in adulthood on the other. METHOD: We used data from 7740 respondents of the European Study of Epidemiology of Mental Disorders project, a cross-sectional household survey carried out in six European countries. The data were assessed with the World Mental Health Composite International Diagnostic Interview, a comprehensive, fully structured psychiatric diagnostic interview. Suicidality was categorized as follows: 'no ideation', 'ideation', 'attempt'. Parental bonding was assessed by means of a three-factor ('care', 'overprotection', 'authoritarianism') short form of the Parental Bonding Instrument. Using a multinomial-logistic regression model to investigate the association between these two constructs, we also adjusted for mood disorders, anxiety disorders, alcohol abuse/dependence and possible country effects. RESULTS: We found associations between low maternal and paternal care on the one hand and suicidality on the other. Country-specific differences proved negligible. CONCLUSIONS: Prevention programs can help better equip parents in their child-rearing role to create a more caring parenting environment. This can be a protective factor for suicidality in adulthood. Nevertheless, more efforts are necessary to better describe the paths that lead from child-rearing behaviour to suicidality in adulthood. PMID- 17464684 TI - Working memory, catecholamines and psychosis: illustrative case. AB - OBJECTIVE: To briefly review the role of catecholamines in prefrontal functions and working memory as illustrated by a case study. METHOD: The work of Goldman Rakic and Arnsten on working memory is briefly reviewed. A case study that illustrates catecholamine functions in an autistic disorder child, who suffered a prolonged psychosis, is described. RESULTS: While the role of dopaminergic neurotransmission in working memory has been described, the present case also illustrates a role for a noradrenergic re-uptake inhibitor in treating the post psychotic distractibility of a severely impaired early adolescent. CONCLUSION: The role of catecholamine neurotransmitters in the treatment of prefrontal symptoms should be further investigated. PMID- 17464685 TI - Neuroscience Institute of Schizophrenia and Allied Disorders (NISAD): 10 years of Australia's first virtual research institute. AB - OBJECTIVE: To review the first 10 years of operation of the Neuroscience Institute of Schizophrenia and Allied Disorders (NISAD), Australia's first virtual research institute. METHOD: Narrative description of the evolution of NISAD. RESULTS: Since inception in 1996, NISAD has developed a wide range of activities to enhance existing efforts and develop new initiatives in schizophrenia research, initially throughout New South Wales, but increasingly on a national scale. This involved the initial development of critical research infrastructure to provide the foundation, with the subsequent focus on developing a multidisciplinary programme of schizophrenia research, across the basic to applied research spectrum. While the primary focus has been the scientific domain, NISAD has also played a leading role in increasing public awareness of schizophrenia as a disease amenable to scientific investigation. CONCLUSION: NISAD has succeeded in building a framework to apply the latest developments in neuroscience to the study of schizophrenia and has formed a multidisciplinary network of clinicians and neuroscientists who are actively collaborating on a range of research initiatives. The 'virtual institute' structure of NISAD has proven cost-efficient and consistent with innovative thinking about research resource management. PMID- 17464686 TI - Bradyarrhythmic shock associated with olanzapine. PMID- 17464688 TI - Status of computerized cognitive behavioural therapy for adults. AB - A growing body of evidence supports the efficacy of computerized cognitive behavioural therapy (CCBT). This technology has the potential to increase the capacity of mental health services, and to overcome some of the barriers to accessing mental health services, including stigma, traveling time for rural patients, treatment delays, and the low availability of skilled clinicians. This review discusses key issues around the implementation of CCBT in current mental health services, and summarizes recent evidence for the efficacy of CCBT in anxiety and depression. Many CCBT systems exist, and the evidence for each varies in quality and quantity. It is concluded that CCBT, particularly guided by a therapist, represents a promising resource. However, considerable work needs to be done to develop CCBT techniques that are appropriate to Australasian populations, acceptable to patients and clinicians, easy to use, and are clinically and cost effective. Suggestions are made for further research and useful website addresses are provided to assist clinicians in familiarizing themselves with CCBT. PMID- 17464689 TI - Neuropsychological impairment in major depression: its nature, origin and clinical significance. AB - Neuropsychological impairment is well established as a feature of major depressive disorder (MDD) but studies have shown a variable pattern of impairment. This paper seeks first to clarify this by examining methodological and clinical factors that give rise to variability in study findings. Second, it examines theories of the origin of these neuropsychological abnormalities. Third, it reviews evidence regarding the clinical significance of different patterns of deficit. A selective review was undertaken of the literature with a particular emphasis on methodological factors, the influence of clinical subtypes and prevalent theories of neuropsychological abnormality. Methodological issues and the heterogeneity of MDD account for considerable variability in results. Specific investigation of the subtypes of psychotic MDD, melancholic MDD and bipolar depression reduces this heterogeneity and results are more consistent in the elderly. Hypothalamic-pituitary-adrenal axis dysfunction is associated with neuropsychological dysfunction in MDD although evidence of direct causation is not definitive at present. Impairment of executive and psychomotor function is a consistent finding, particularly in the elderly, and may reflect frontostriatal limbic dysfunction. There is growing evidence that this may have clinical significance. It is suggested that future research take very careful account of the exact phenotype of MDD. Classification based on neuropsychological profile may, in fact, be useful. Further research should examine further the clinical importance of patterns of neuropsychological impairment. PMID- 17464690 TI - Increased prefrontal cerebral blood flow in first-episode schizophrenia following treatment: longitudinal positron emission tomography study. AB - OBJECTIVE: Cognitive deficits, particularly those related to executive function and behavioural control, are a core feature of patients with schizophrenia and implicate disturbances of the prefrontal cortex (PFC). Consistent with this, functional imaging studies have identified abnormalities of PFC activity in chronically affected patients. The objective of the current study was to investigate executive-control related neural activity from first onset of the illness through to symptom stabilization. METHOD: The authors examined eight neuroleptic-naive first-episode psychosis (FEP) patients within 3 days of first presentation and eight healthy age- premorbid-IQ, and gender-matched controls (CTL). All FEP patients were later confirmed with a diagnosis of schizophrenia. Subjects underwent H2-15O positron emission tomography (PET) while performing the Stroop interference task that has previously been shown to engage the PFC in healthy samples. In a double-blind paradigm, FEP patients were randomly treated with either haloperidol or risperidone for 8 weeks, after which CTLs and all but one of the FEP patients were re-scanned. RESULTS: Behaviorally, there was no change in task performance from baseline to follow up for either the FEPs or CTLs. However, there were significant changes in functional activation in both groups across the same period. For CTLs, task-performance was associated with greater recruitment of posterior brain regions at follow up compared to baseline, while for FEP this involved greater recruitment frontal regions. Concurrently, FEP also showed significant improvement in positive symptomatology. CONCLUSIONS: These findings suggest that disturbances of the PFC often seen in FEP may be, at least partially, associated with acute symptom expression. However, it is still unclear whether this increase in frontal activity is due to symptom stabilization per se, the effects of medication, a lack of neurophysiological 'learning' with experience/practice, or a combination of these factors. In the context of the CTLs performance, we interpret the patient findings as reflective of greater neurophysiological effort required for task accomplishment relative to the learned, more automated, processing apparent in the CTLs. PMID- 17464691 TI - Sex differences in the cerebral function associated with processing of aversive stimuli by schizophrenia patients. AB - OBJECTIVE: Impaired processing of various emotions is considered one of the fundamental features of schizophrenia. In the recent study intriguing sex differences were observed in the cerebral function associated with the experience of sadness in schizophrenia patients. The aim of the present study was to explore this phenomenon during exposure to aversive stimuli. METHOD: Fifteen men and 10 women with the DSM-IV diagnosis of schizophrenia underwent functional magnetic resonance imaging (fMRI) while viewing alternating blocks of negative and neutral pictures. Data were analysed using random-effects model within statistical parametric mapping (SPM99) software. RESULTS: Processing of negative stimuli evoked significantly greater activations in men in the thalamus, cerebellum, temporal, occipital and posterior cingulate cortex, while women exhibited greater activations in the left middle frontal gyrus. CONCLUSIONS: The sex differences in the cerebral activations in schizophrenia patients deviate from what has been observed in the general population during exposure and experience of negative affect. As such the present study supports and extends the authors' preliminary observation of the anomalous sexual dimorphism in schizophrenia at the functional neuroanatomical level, suggesting potential masculinization of female subjects and feminization of male subjects with schizophrenia. PMID- 17464692 TI - Better outcomes in mental health care: impact of different models of psychological service provision on patient outcomes. AB - OBJECTIVE: One hundred and eight Access to Allied Psychological Services projects have been funded under Australia's Better Outcomes in Mental Health Care programme since July 2001. All projects are run by Divisions of General Practice and enable general practitioners (GPs) to refer patients to allied health professionals for evidence-based care. They differ in the models they use to retain, locate and direct referrals to their allied health professionals. This paper examines the extent to which the projects are achieving positive patient outcomes, and explores the association between different models of service delivery and varying levels of patient outcomes. METHOD: The paper draws on two data sources (a purpose-designed minimum dataset and a survey of models of service delivery) to examine the level of patient outcomes within and across projects, and variations in the level of patient outcomes by models of service delivery. RESULTS: The projects are achieving positive effects and these are mostly of large or medium magnitude. The projects do not differ markedly in terms of the patient outcomes they are achieving, despite differences in the models of service delivery they are using. However, those projects implementing a direct referral model, where the GP refers the patient directly to the allied health professional, have significantly greater effect sizes, indicating that they are achieving greater improvements in patient outcomes. In addition, there are non significant trends toward direct employment of allied health professionals by Divisions being predictive of greater improvements in patient outcomes, and delivery of services from allied health professionals' own rooms being predictive of weaker patient outcomes. CONCLUSIONS: Overwhelmingly, the Access to Allied Psychological Services projects are having a positive impact for patients in terms of their level of functioning, severity of symptoms and/or quality of life. Preliminary indications suggest that a service delivery model incorporating the use of a direct referral system may be associated with superior outcomes. The findings are discussed in the light of the imminent listing of psychologists' services on the Medicare Benefits Schedule. PMID- 17464693 TI - Mother and baby unit admissions: feasibility study examining child outcomes 4-6 years on. AB - OBJECTIVE: To compare the cognitive, social cognitive, behavioural, emotional and attachment security outcomes of children whose mothers were admitted to a psychiatric mother and baby unit (MBU) with those of offspring of mothers not admitted and with standardized norms. METHOD: Children aged 4-6 years of mothers admitted (following childbirth) to the dedicated psychiatric MBUs in Manchester, and still living with their mother, were traced for recruitment. Observational and multi-informant measures of developmental functioning were administered during a home visit. RESULTS: From 94 consecutive admissions, 16 children participated and showed no poorer developmental outcomes than comparison children or standardized child norms. Girls, securely attached children and children of mothers with no current mental illness tended to show the most positive outcomes. CONCLUSION: Possible reasons for low participant recruitment are discussed. Our provisional findings suggest that interventions should focus on improving infant attachment security, targeted at women with psychotic and chronic illness, who have no partner and/or who have a male child. PMID- 17464694 TI - New Zealand psychiatrists views on global features of ICD-10 and DSM-IV. AB - OBJECTIVE: The present accepted classificatory systems (ICD-10 and DSM-IV) represent the culmination of 100 years of post-Kraepelinian evolution. The present paper reports on a study to ascertain the views of New Zealand psychiatrists on their requirements of a classificatory system, and their opinions on those currently in use. METHOD: An anonymous postal survey of 542 psychiatrists and trainees was undertaken in New Zealand. RESULTS: A total of 235 questionnaires were returned, completed. New Zealand psychiatrists primarily use DSM-IV and do so because that is what they were taught on. They make relatively limited use of the multiaxiality. The purposes of classificatory systems that they value most highly are reliable interclinician communication, and to inform patient management planning. The two purposes they valued least were usefulness for a national statistical base, or to indicate prognosis. CONCLUSIONS: New Zealand psychiatrists' views are consistent with some of the stated objectives of ICD-10 and DSM-IV, but there is significant diversity in the former and over ambition in the latter, with much to be resolved. PMID- 17464695 TI - Reliability and validity of the Kessler 10 and Patient Health Questionnaire among injecting drug users. AB - OBJECTIVE: To examine the reliability and validity of the Kessler 10 (K10) and the Patient Health Questionnaire (PHQ) in a sample of injecting drug users (IDUs). METHOD: Participants were 103 IDUs with a current substance use disorder accessing a needle and syringe programme. Presence of mental health disorders was assessed using the Mini International Neuropsychiatric Interview (MINI). RESULTS: Both the K10 and PHQ had high levels of internal consistency and concurrent validity. Individuals with a positive screen on the K10 were ten lines more likely to have a current affective disorder, while those with a positive PHQ screen had nearly 14 times the risk. CONCLUSIONS: The K10 and PHQ are recommended as brief screening and diagnostic tools for current affective disorders among IDUs. PMID- 17464696 TI - AKT1 and neurocognition in schizophrenia. AB - OBJECTIVE: Previous research has shown conflicting results for the significance of five v-akt murine thymoma viral oncogene homolog 1 (AKT1) single-nucleotide polymorphisms (SNPs) to the aetiology of schizophrenia. Neurocognition is a plausible endophenotype for schizophrenia and it was reasoned that the lack of agreement might be due to variability in neurocognition across studies. Therefore, the association of genetic variation in AKT1 with neurocognition was investigated in patients with schizophrenia. METHODS: The same five SNPs used in previous studies of the etiology of schizophrenia (rs2494732, rs2498799, rs3730358, rs1130214, [corrected] and rs3803300) were genotyped in 641 individuals with schizophrenia who had participated in the Clinical Antipsychotic Trials of Intervention Effectiveness (CATIE) project. The primary dependent variable was a neurocognitive composite score and exploratory analyses investigated five domain scores (processing speed, reasoning, verbal memory, working memory, and vigilance). RESULTS: There were no significant asymptotic or empirical associations between any SNP and the neurocognitive composite score. The authors also investigated the association of five-SNP haplotypes with the neurocognitive composite score. A marginally significant association was observed for the neurocognitive composite score with one of the five-SNP haplotypes (global score statistic 19.51, df = 9, permutation p = 0.02). Exploratory analyses of five domain scores (processing speed, reasoning, verbal memory, working memory, and vigilance) were non-significant for all five SNPs. CONCLUSION: Results published to date for an association between genetic variation in AKT1 with schizophrenia are inconsistent. The results suggest that the AKT1 markers studied are not associated with neurocognition in schizophrenia, and do not support unassessed variation in neurocognitive scores as a reason for this discrepancy. PMID- 17464697 TI - Lundby revisited: first incidence of mental disorders 1947-1997. AB - OBJECTIVE: To investigate how first incidence of various mental disorders changed between the periods of 1947-1972 to 1972-1997 in the Lundby cohort. METHOD: First incidence rates of mental disorders were calculated for two 25 year periods and ten 5 year periods. RESULTS: From 1947-1972 to 1972-1997 a decrease in almost all age- and sex-specific incidences of neurotic and organic brain disorders was observed, whereas incidence rates of psychotic disorders increased consistently in male subjects but decreased in most age intervals in female subjects. For both sexes the age-standardized 5 year period incidences of neurotic disorders decreased after 1972, fluctuated for psychotic disorders 1947-1997 and decreased steadily for organic disorders 1947-1997. CONCLUSIONS: The reduction in neurotic and organic brain disorder incidences may be linked to structural changes in society and medical advances. PMID- 17464698 TI - Zero tolerance approach to aggression and its impact upon mental health staff attitudes. AB - OBJECTIVE: A natural study design provided an opportunity to investigate the effect of zero tolerance to aggression policy on health staff attitudes. METHOD: This study compared a training programme used to train health service staff in aggression minimization (group 1) with a highly similar training programme that differed in that it emphasized a zero tolerance approach to aggression minimization in the health workplace (group 2). Prior to training and at follow up 117 health staff (57 from group 1, and 60 from group 2) completed a series of evaluations examining rigid attitudes toward managing aggression, tolerance for aggression and confidence and skills in dealing with aggression. RESULTS: The groups were equivalent with respect to age, sex and occupation. Prior to training the two groups did not differ with regard to rigidity, tolerance, confidence and skills. Both training interventions were effective in increasing confidence and skills. However, group 1 significantly decreased rigid attitudes toward the management of aggression, whereas group 2 significantly evidenced increased rigid attitudes and decreased tolerance toward aggression. CONCLUSION: Zero tolerance for aggression had a negative impact on health staff attitudes in relation to managing aggression in the health workplace. PMID- 17464699 TI - Prolongation of clozapine-induced neutropenia with olanzapine. PMID- 17464701 TI - Benefits of cognitive-behavioural therapy for children and youth with obsessive compulsive disorder: re-examination of the evidence. AB - The aim of the present paper was to critically examine evidence about the benefits of cognitive-behavioural therapy (CBT) for pediatric obsessive compulsive disorder (OCD) from controlled and single group studies, including its benefits relative to medication are critically reviewed. Selected studies were categorized by study type and by risk of bias classification. Standardized mean differences (Hedges' g or Cohen d) and, where appropriate, weighted mean difference (WMD) were calculated. All five comparison and 14 one-group studies showed a significant benefit for CBT within a wide range (ES = .78 to 4.38). Low risk of bias studies produced the lower adjusted effect sizes. The best available estimate of CBT efficacy relative to no treatment is about 1 standardized mean difference, equivalent to a treatment effect of 8 points on the Children's Yale Brown Obsessive-Compulsive Scale. This represents a reduction in the risk of continuing to have OCD post-treatment of about 37% (95% CI 14% to 54%). Evidence from 3 studies indicates that the efficacy of CBT and medication do not differ significantly. CBT combined with medication is significantly more efficacious than non-active controls or medication alone but not relative to CBT alone. CBT should be regarded as a first line equivalent to anti-OCD medication with the potential to lead to better outcomes when combined with medication than medication alone can provide. Additional studies are needed to further clarify CBT's benefits and to investigate how it can be made more available as a treatment option for children and youth who suffer from OCD. PMID- 17464702 TI - Self-reported suicide attempts and associated risk and protective factors among secondary school students in New Zealand. AB - OBJECTIVE: To examine associations between individual, family, school and community characteristics and rates of suicide attempts in a national population sample of New Zealand secondary school students. METHOD: A total of 9570 randomly selected 9- to 13-year-old students from 114 schools were surveyed, using the New Zealand Adolescent Health Survey. This is a 523-item anonymous self-report comprehensive questionnaire delivered by Multi-Media Computer-Assisted Self Interviewing. Multivariate analyses were used to examine correlates of self reported suicide attempts within the last 12 months. RESULTS: In total, 739 participants (4.7% of males and 10.5% of females) reported having made a suicide attempt within the last 12 months. Depressive symptoms, alcohol abuse, -having a friend or family member attempt suicide, family violence and non-heterosexual attractions were independently associated with increased rates of suicide attempts while parents caring, other family members caring, teachers being fair and feeling safe at school were independently associated with decreased rates of suicide attempts. Caring friendships, attending worship frequently, possible sexual abuse and anxiety symptoms were not independently associated with suicide attempts. Risk and protective factors operated in the same way for male and female students and for those with and without other suicide predictors. CONCLUSIONS: New Zealand secondary school students, particularly female students, report high rates of suicide attempts. Risk of suicide attempts is lower in students reporting caring home and fair, safe school environments and this effect remains once depression is taken into account. This study confirms the importance of depression, substance use, problem behaviour, negative life events, exposure to suicide behaviour by others and the significance of sexual orientation in suicidal behaviour among school students and provides evidence of the importance of the family and school environments in reducing risk among this group. PMID- 17464703 TI - Atomoxetine versus methylphenidate in paediatric outpatients with attention deficit hyperactivity disorder: a randomized, double-blind comparison trial. AB - OBJECTIVE: To (i) test whether atomoxetine is non-inferior to methylphenidate in treating symptoms of attention deficit hyperactivity disorder (ADHD) in paediatric patients; and (ii) determine the tolerability of the two drugs. METHOD: This double-blind study was conducted in 6- to 16-year-old outpatients with ADHD (DSM-IV) in China, Korea and Mexico (January-October 2004). Patients were randomly assigned to once-daily atomoxetine (0.8-1.8 mg kg(-1) day(-1); n = 164) or twice-daily methylphenidate (0.2-0.6 mg kg(-1) day(-1); n = 166) for approximately 8 weeks. Primary efficacy assessment was the comparison of response rates (> or =40% reduction from baseline to end point in total score) on the Attention Deficit Hyperactivity Disorder Rating Scale-IV-Parent Version: Investigator-Administered and -Scored. Tolerability measures included, but were not limited to, the assessment of treatment-emergent adverse events (TEAEs) and weight. RESULTS: Atomoxetine was non-inferior to methylphenidate in improving ADHD symptoms based on response rates (atomoxetine, 77.4%; methylphenidate, 81.5%; one-sided 95% lower confidence limit = -11.7%, p = 0.404). Treatment emergent adverse effects experienced significantly more frequently in the atomoxetine group, compared with the methylphenidate group, included anorexia (37.2% vs. 25.3%; p = 0.024), nausea (20.1% vs. 10.2%; p = 0.014), somnolence (26.2% vs. 3.6%; p <0.001), dizziness (15.2% vs. 7.2%; p = 0.024) and vomiting (11.6% vs. 3.6%; p = 0.007), most of which were of mild or moderate severity. Atomoxetine-treated patients experienced a small but significantly greater mean weight loss from baseline to end point than methylphenidate-treated patients ( 1.2 kg vs. -0.4 kg; p <0.001). CONCLUSIONS: This study suggests that atomoxetine is non-inferior to methylphenidate in the improvement of ADHD symptoms in paediatric outpatients. Although both of the drugs were well tolerated, atomoxetine was associated with a higher incidence of TEAEs than methylphenidate. PMID- 17464704 TI - Problem behaviours in early primary school children: Australian normative data using the Strengths and Difficulties Questionnaire. AB - OBJECTIVE: This study provides normative and clinical levels of problem behaviours from a large community sample of Australian early primary school children. METHOD: From a large community sample (n = 1928) of children aged 5 to 10 years (mean = 7.70, SD = 0.89) normative data are provided using the teacher reported version of the Strengths and Difficulties Questionnaire (SDQ). A response rate of 88% ensures the data are representative. RESULTS: Mean scores on total difficulties between this Australian sample and UK norms were comparable. Some differences in the clinical cut-off scores are evident in the Australian sample, and adjusted clinical cut-offs are provided. Australian teacher reports of problem behaviour revealed lower clinical levels of emotional problems. Boys had significantly greater problems than girls on the Conduct, Hyperactivity, Peer and Prosocial scales. Overall, 5.3% of boys had scores in the clinical range on four of the five subscales. Comparisons of the factor structure revealed that for boys the Peer scale has two interpretations, with two items loading with Conduct problems and associated with greater problem behaviours. For girls, conduct problems are more strongly associated with poorer prosocial skills. CONCLUSIONS: Teachers are able to provide valuable predictive information on externalizing behaviours. Australian normative comparisons reveal minor variations in teacher interpretation of items. The present data is representative of the Australian community and should be used to assess behaviour difficulties in early primary school children. Future studies on older age children are required to understand the developmental progression of problem behaviours in the community. PMID- 17464705 TI - Does the triple P-Positive Parenting Program provide value for money? AB - OBJECTIVE: The aim of the present study was to investigate the economic case for the implementation of the Triple P-Positive Parenting Program on a population basis in Queensland, Australia, in order to reduce the prevalence of conduct disorder in children. METHOD: Threshold analysis was undertaken together with a limited cost-effectiveness analysis. RESULTS: The Triple P-Positive Parenting Program is a dominant intervention; that is, it costs less than the amount it saves, until the reduction in prevalence falls below 7% where net costs become positive. CONCLUSIONS: Triple P is likely to be a worthwhile use of limited health funds. The economic case is promising, but further research is required to confirm the study results. PMID- 17464706 TI - Predictors of attitudes towards treatments for bulimia nervosa. AB - OBJECTIVE: The aim of this study was to examine, using Mental Health Literacy, acceptability and correlates of acceptability of three treatment interventions for bulimia nervosa; medication, psychological therapy and self-change/lifestyle therapies. METHOD: A self-report questionnaire to ascertain correlates of attitudes towards the three interventions was used. Respondents (n = 177) were females aged 18-53 years. RESULTS: Psychological therapy was rated the most acceptable treatment to participants as well as the most acceptable to others, and to have most advantages and fewest disadvantages, followed by self change/lifestyle therapies, and then medication. Social Pressures to be Thin was the most strongly endorsed cause of bulimia, followed by Low Self-Esteem, Eating Behaviour, Relationship and Family Environment, and lastly, Biology. Correlates with acceptability of different treatment approaches were identified. CONCLUSIONS: Although no causal link can be assumed from this design, findings suggest that women appear to have a mixed understanding of the different mechanisms that each treatment intervention adopts to treat the causes and symptoms of bulimia, particularly self-change/lifestyle therapies. This may help to explain poor adherence to interventions. PMID- 17464707 TI - Risk of repetition of suicide attempt, suicide or all deaths after an episode of attempted suicide: a register-based survival analysis. AB - OBJECTIVE: This study was undertaken in order to estimate the incidence of repetition of suicide attempt, suicide and all deaths, and to analyse the influence of psychiatric illness and socio-demographic factors on these. METHOD: The study is a Danish register-based survival analysis that retrieved personal data on socio-economic, psychiatric and mortality conditions from various registers. Hazards were estimated using Cox regression with a time-dependence covariate. Suicide-attempters (2.614) and non-attempters (39.210)were analysed being matched by gender, age and place of residence. RESULTS: The average follow up period for suicide-attempters was 3.88 years, during which 271 (10.37%) of them died. By comparison, death occurred four times more often among suicide attempters than among non-attempters. Suicide was far more common among attempters (61, 2.33%) than among non-attempters (16, 0.04%). A proportion of the attempters (31.33%) repeated their attempt within the follow-up period. The most reliable predictors for suicide and death were repetition, suicide attempt method and treatment for mental illness. The most reliable predictors for repetition were age, gender and mental illness. DISCUSSION: Individuals with a history of suicide attempts form a well-defined high-risk group for suicide, and are in need of treatment immediately after the episode. Somatic and psychiatric staff must be informed about the risk factors for subsequent suicidal behaviour after an episode of attempted suicide. Furthermore, departments that are in contact with suicidal individuals need action plans to ensure that all such individuals are discharged to proper treatment immediately after the suicide attempt. PMID- 17464708 TI - Levels of serum interleukin (IL)-6, IL-1beta, tumour necrosis factor-alpha and leptin and their correlation in depression. AB - OBJECTIVE: To investigate the relationship between leptin and cytokines in depressed patients. METHODS: Thirty-three unmedicated patients (24 female, nine male) with depressive disorder and 23 healthy controls (16 female, seven male) were assessed for serum levels of interleukin (IL)-6, IL-1beta, tumour necrosis factor-alpha (TNF-alpha) and leptin. RESULTS: Levels of IL-6 and TNF-alpha in depressed patients were higher than in normal controls. There were significantly lower leptin levels in depressed patients than in normal controls. There were also significant differences in the leptin levels, being higher in female than in male patients, and in female than in male controls. CONCLUSIONS: IL-6 and TNF alpha cytokines and leptin are important in the psychoimmunology of depressed patients. There were gender differences in leptin levels in depression. PMID- 17464709 TI - Mortality associated with incident mental health disorders after stroke. AB - OBJECTIVE: Sparse information is currently available about the incidence of the major psychiatric syndromes following a stroke and their long-term contribution to morbidity and mortality. This study was designed to determine: (1) the incidence of first ever mental health disorder in amongst stroke patients; (2) the 10-year mortality associated with incident post-stroke mental health disorders. METHODS: DESIGN: Retrospective cohort study. SETTING: Entire Western Australian community. PARTICIPANTS: First-ever stroke in 1990. Subjects with a prior recorded history of any mental health disorder were excluded from the study. Main outcomes of interest: Incident mental health diagnoses and 10-year mortality. RESULTS: 1,129 hospital stroke contacts were recorded in 1990, with 21 people dying on the same day of contact. Between 1990-2002 36.6% of the survivors received a mental health diagnosis (6.1 per 1,000 person-years): alcohol-related disorders (16.2%), dementia (12.1%), delirium (7.6%), psychotic disorders (6.7%), and depression (5.5%). Mental health disorder onset was usually within 6 months of the stroke. Patients with an incident psychotic disorder were twice as likely to die during the subsequent 10 years as post-stroke controls with no mental health disorder (risk ratio = 2.03, 95%CI = 1.39-2.95). Being a widow (HR = 1.61, 95%CI = 1.13-2.30) or having been born in 'other countries' as opposed to Australia (HR = 1.56, 95%CI = 1.15-2.11) was also associated with increased death hazard. CONCLUSIONS: Approximately 1 in 3 patients develop a mental health disorder after stroke, although incidence estimates are relatively low. Post stroke psychosis is associated with greater 10-year mortality, but the mechanisms underlying such an association are yet to be determined. PMID- 17464710 TI - Coping and resilience in refugees from the Sudan: a narrative account. AB - OBJECTIVE: The purpose of this paper was to identify and explicate coping and resilience themes employed by 13 re-settled Sudanese refugees. METHOD: A sample of 13 Sudanese refugees was asked to describe their experience of coping in the three periods of their migration, namely: pre migration from Sudan, transit, and post migration in their host country. Members of the sample participated in qualitative interviews conducted with the assistance a bilingual community worker. RESULTS: Three themes that characterized the experience of re-settled refugees across all periods were: religious beliefs, social support and personal qualities. A fourth less salient, theme, comparison with others, also emerged in the post-migration context. CONCLUSIONS: A number of themes associated with coping and resilience in response to trauma were identified. These themes may be translated into strategies to assist in responding constructively to trauma. Such approaches may be used to improve the well-being of re-settled refugees in Australia. PMID- 17464711 TI - Managing psychosis in pregnancy. AB - OBJECTIVE: This article provides an introduction to the complex issues surrounding the management of women who have a history of psychosis and who become pregnant. Balancing the mental wellbeing of the woman and the safety and wellbeing of the baby is a complex task for both the expectant mother and the health professionals involved in her care. CLINICAL PICTURE: Within this article the complexity of the issues will be outlined as a case report of a woman with a history of psychotic related disorders, who was also pregnant. TREATMENT: The woman was being case managed by a Mental Health Service in Victoria, Australia, and was included on the National Register of Antipsychotic Medications in Pregnancy Register (NRAMP) recently established at the Alfred Psychiatry Research Centre (APRC). OUTCOME: The profile of women with a history of previous mental illness, and who are pregnant, often includes a poor psychosocial history and involvement with child protection agencies with regard to custody of the children. Well meant but poorly coordinated decisions by health professionals result in sub-optimal outcomes for both mother and infant. CONCLUSION: There is a need for the exploration of the management and experiences of women who have a history of psychosis and who are pregnant. This case example highlights the complexity of issues surrounding the management of this vulnerable group of women and their babies. PMID- 17464712 TI - Management of clozapine-induced obsessive-compulsive symptoms in a man with schizophrenia. PMID- 17464713 TI - Self-cannibalism: an unusual case of self-mutilation. PMID- 17464714 TI - Preventing suicide among depressed patients in primary care practice. PMID- 17464716 TI - Are violence risk assessment tools clinically useful? AB - Despite increasing concerns regarding the prevalence of violent behaviour in mainstream mental health settings, the impressive body of forensic research on violence risk assessment has thus far had only limited impact on front-line general mental health practice. The common objection raised by clinicians that risk assessment tools lack utility for clinical practice may contribute to this. The present paper argues that this objection, although understandable, is misplaced. Usage of appropriate, validated risk assessment tools can augment standard clinical approaches in a number of ways. Some of their advantages derive simply from having a well-structured approach, others from consideration of specific kinds of risk factors: 'static' and 'dynamic'. The inappropriate use of tools without a firm evidence base, however, is unlikely to enhance clinical practice significantly. PMID- 17464717 TI - Gene expression profiling in Brodmann's area 46 from subjects with schizophrenia. AB - OBJECTIVE: To identify altered gene expression in the dorsolateral prefrontal cortex obtained after death from subjects with schizophrenia. METHOD: Restriction fragment differential display (RFDD) was used to measure levels of mRNA in Brodmann area (BA) 46 from schizophrenia and control subjects. Findings on specific mRNA identified with RFDD were further investigated using real-time polymerase chain reaction (real-time PCR), PCR and western blotting. RESULTS: Levels of mRNA for 63 of approximately 12,500 genes differed in BA 46 in schizophrenia. Subsequent real-time PCR has shown that mRNA for muscleblind protein 1 (MBNL1) and protocadherin 17 (PCDH17) are increased in BA 46 from subjects with schizophrenia of short, but not long, duration. Altered levels of mRNA for neither gene were present in BA 9 from subjects with schizophrenia or in either cortical area from subjects with bipolar 1 disorder. By contrast, both RFDD and real-time PCR failed to show altered expression of the schizophrenia candidate gene disrupted in schizophrenia 1 (DISC1) BA46 from any diagnostic cohort. CONCLUSION: The present study has identified genes that are differentially expressed in BA 46 in schizophrenia. Initial studies have shown that there is a need for a careful validation of genes shown to be affected in schizophrenia using high-throughput technologies. In addition the present study has shown that gene expression may vary considerably depending on the duration of schizophrenia. This raises the hypothesis that changing gene expression may be underlying the change in symptom profile that occurs with disease progression in some subjects with schizophrenia. PMID- 17464718 TI - Changes in brain-derived neurotrophic factor following treatment with mifepristone in bipolar disorder and schizophrenia. AB - OBJECTIVE: Brain-derived neurotrophic factor (BDNF) is stress-responsive and has been implicated in a number of disparate neuropsychiatric disorders. Glucocorticoid antagonists have been shown to have beneficial effects on mood and cognitive function in bipolar disorder but not in schizophrenia. The aim of the present study was to investigate BDNF levels in patients with bipolar disorder and schizophrenia before and after treatment with the glucocorticoid receptor antagonist mifepristone. METHODS: Peripheral BDNF levels were measured in patients with bipolar disorder (n=20), schizophrenia (n=20) and 14 matched healthy controls following 7 days of adjunctive mifepristone (600 mg day(-1)) treatment in a double-blind, placebo-controlled crossover design study. RESULTS: Baseline BDNF values were similar in both patient groups and in healthy controls. Following treatment with mifepristone, cortisol levels were significantly increased and BDNF levels decreased in both schizophrenia and bipolar disorder. A significant correlation existed between change in cortisol level and change in BDNF levels following mifepristone treatment in schizophrenia, but not in bipolar disorder. CONCLUSION: Differing BDNF responses to increasing cortisol levels between patients with schizophrenia and with bipolar disorder may reflect underlying pathophysiological mechanisms. PMID- 17464719 TI - Regional gray matter changes in bipolar disorder: a voxel-based morphometric study. AB - OBJECTIVE: To investigate structural abnormalities in bipolar disorder (BD) using optimized voxel-based morphometry (VBM) in closely matched patients and controls, and to examine the relationship of clinical features with regional gray matter (GM) volumes. METHODS: Twenty-four patients (six male) aged 19-59 years (mean=38.21 years, SD=11.04 years) with DSM-IV bipolar I disorder were compared with 25 control subjects, matched on age, sex, and years of education. VBM analyses were conducted on high-resolution T1-weighted brain magnetic resonance imaging to detect regional GM volume differences between groups, ensuring statistical correlation for age, sex and total intracranial volumes. Within the patient groups, regional GM changes were also investigated. RESULTS: Compared to controls, BD patients had increased GM volume in left parahippocampal gyrus and decreased GM volume in left middle temporal gyrus. Family history, psychotic symptoms and lithium status were associated with regional GM abnormalities in BD patients. CONCLUSIONS: This study presents evidence of gray matter volume abnormalities in adults with bipolar I disorder. Regional variation in relation to clinical factors suggests a neurobiological basis for clinical heterogeneity and posits the possibility of trait deficits. PMID- 17464720 TI - Reassessing the aftercare treatment of individuals found not guilty due to a mental disability in Hunan, China: supplemental study into the disposition of mentally ill offenders after forensic psychiatric assessment. AB - OBJECTIVE: The purpose of the present paper was to examine the disposition of individuals in Hunan, China who are found not criminally responsible due to a mental disorder or defect. METHOD: Self-developed questionnaires were mailed to the family members of 240 patients who had received forensic psychiatric evaluations at the forensic psychiatric assessment center of Central South University, between 2001 and 2002. RESULTS: One hundred and seventy questionnaires were fully completed and returned by the patients' family members. According to the answers, 64.1% (109 patients) were found not criminally responsible due to a mental disorder or defect. In 87.6% of the cases, a judgement of guilty was in agreement with the psychiatrist's recommendation concerning criminal responsibility. A total of 61.8% of the patients found not guilty were discharged to their families and did not receive any further psychiatric treatment. CONCLUSION: In most cases, judicial decisions are consistent with a psychiatrist's opinion of criminal responsibility due to a mental disorder or defect. After such adjudication, further psychiatric treatment is often neglected due to the lack of resources and information. Hunan, China must make a continued investment into the availability and quality of outpatient mental health treatment for forensic patients after they have been discharged. PMID- 17464721 TI - Inpatient psychosocial rehabilitation in rural NSW: assessment of clinically significant change for people with severe mental illness. AB - OBJECTIVE: The aim of the present study was to describe an inpatient psychosocial rehabilitation programme in rural New South Wales and to assess the effectiveness of the programme using measures of clinically significant change. METHOD: The first 88 patients with a schizophrenia spectrum disorder to enter the Manara Clinic and Turon House, New South Wales, psychosocial rehabilitation programmes were assessed at admission and discharge using the Brief Psychiatric Rating Scale, the Health of the Nation Outcome Scales, and the Kessler-10 self-report measure. RESULTS: Significant improvements in psychiatric symptomatology, psychosocial functioning, and psychological distress were found over the course of the inpatient stay. Clinical significance analyses using patients in the community as the reference group indicated a reliable and clinically significant improvement for 33% of inpatients on psychiatric symptomatology, 39% of inpatients on psychosocial functioning, and 21% of inpatients on psychological distress. CONCLUSIONS: The psychosocial rehabilitation programme provides clinically significant initial benefits for patients with severe mental illnesses. More attention needs to be paid to evaluating which components of psychosocial rehabilitation contribute most to these benefits. Follow-up evaluation is required to determine whether the benefits of this programme are sustained in the community. PMID- 17464722 TI - Eating disorders across two cultures: does the expression of psychological control vary? AB - OBJECTIVE: Because both the expression of eating disorder (ED) symptoms and preferred psychological control styles may be affected by culture, the purpose of the present study was to examine whether the expression of psychological control in women with EDs varies across two cultures. METHOD: North European Australian and Chinese Singaporean women (n = 117) with anorexia nervosa (n = 36), bulimia nervosa (n = 13) and eating disorders not otherwise specified (n = 3), and without an ED (n = 65) recruited in Australia and Singapore completed a multidimensional inventory assessing sense of control, domains of control, preferred means by which to gain control, and motivation for control. RESULTS: Although the normative control profile for each culture differed slightly, control profiles among those with an ED were very similar across both cultures. However, the directionality and extent of specific aspects of control pathology associated with the presence of an ED differed across cultures. North European Australians with an ED were much more deviant from the cultural norm than their Chinese Singaporean ED counterparts in relation to overall sense of control, methods of gaining control, and control in the domain of body. Chinese Singaporean woman with an ED were much more deviant from the cultural norm than their North European Australian ED counterparts in the domain of control over impulses. CONCLUSIONS: Having an ED powerfully distorts psychological control irrespective of culture. However the degree, directionality, and form of the displacement from normal control styles is also culture dependent. This has implications for treatments that attempt to redress or correct control issues in people with an ED in other cultures. PMID- 17464723 TI - Did geographical inequalities in suicide among men aged 15-44 in New Zealand increase during the period 1980-2001? AB - OBJECTIVE: It has previously been reported that rates of suicide among men aged 15-44 increased rapidly during the 1980s and 1990s. The present study examines whether geographical inequalities in suicide rates among men in this age group rose during the period 1980-2001 in New Zealand. METHODS: Age-standardized rates of suicide for men aged 15-44 were calculated for District Health Boards (DHBs) and in deprivation quintiles (using the 2001 New Zealand deprivation score) for the periods 1980-1982, 1985-1987, 1990-1992, 1995-1997 and 1999-2001. Geographical inequality was measured by calculating the ratio of the suicide rate deprivation quintile 5 to quintile 1. RESULTS: Age-standardized rates of suicide among men aged 15-44 increased in all but two DHBs between 1980 and 2001. The ratio of inequality in suicide between the least deprived and most deprived areas of New Zealand rose from 1.68 in 1980-1982 to a high of 1.94 in 1990-1992, followed by a small reduction to 1.86 by the end of the study period. CONCLUSIONS: Geographical inequalities in suicide among men aged 15-44 grew during the period 1980-2001 and remain at high levels. The greatest increase in inequality was during the late 1980s and early 1990s, a period of rapid social and economic change to New Zealand society. PMID- 17464724 TI - Antidepressants: prevalence of duplicate therapy and avoidable drug interactions in Australian veterans. AB - OBJECTIVE: This study assessed the prevalence of duplicate antidepressant prescribing and avoidable potential antidepressant drug interactions in the Australian war veteran population. METHOD: The Department of Veterans' Affairs Pharmacy Claims database was interrogated, using specific criteria, to identify antidepressant duplication. In addition, potential drug interactions where safer alternative therapies were available were assessed. These included anticholinergic agents with tricyclic antidepressants and tramadol with antidepressants. Episodic tramadol dispensings with antidepressants were also assessed. RESULTS: A total of 46,859 veterans had antidepressants regularly dispensed to them in the period 1 April-31 July 2005. Overall, 4037 potential interactions were identified in 3818 veterans (8.1%) to whom were dispensed antidepressants regularly. Antidepressant and tramadol co-prescribing was the most common potential interaction identified among 3.6% of veterans. Two or more interactions were identified in 212 veterans (0.5%). Analysis of episodic tramadol dispensings with antidepressants suggested a much higher prevalence of 7.7%. CONCLUSION: The increasing use of antidepressants and the high level of potentially avoidable interactions detected in the present study, highlight the necessity of ongoing vigilance concerning the use of potentially inappropriate drug combinations, particularly in the elderly. PMID- 17464725 TI - Citation superstars in psychiatry. PMID- 17464728 TI - Traumatic entrapment, appeasement and complex post-traumatic stress disorder: evolutionary perspectives of hostage reactions, domestic abuse and the Stockholm syndrome. AB - Evolutionary theory and cross-species comparisons are explored to shed new insights into behavioural responses to traumatic entrapment, examining their relationships to the Stockholm syndrome (a specific response to traumatic entrapment) and complex post-traumatic stress disorder (PTSD). A selective literature review is undertaken examining responses to traumatic entrapment (including hostage, domestic abuse and similar situations) and the Stockholm syndrome, before examining mammalian, reptilian and other defensive responses to relevant threats. Chimpanzees, the closest relatives of humans, are closely examined from this perspective and commonalities in behavioural responses are highlighted. The neurobiological basis of defensive behaviours underlying PTSD is explored with reference to the triune brain model. Victims of protracted traumatic entrapment under certain circumstances may display the Stockholm syndrome, which involves paradoxically positive relationships with their oppressors that may persist beyond release. Similar responses are observed in many mammalian species, especially primates. Ethological concepts including dominance hierarchies, reverted escape, de-escalation and conditional reconciliation appear relevant and are illustrated. These phenomena are commonly encountered in victims of severe abuse and understanding these concepts may assist clinical management. Appeasement is the mammalian defence most relevant to the survival challenge presented by traumatic entrapment and appears to be the foundation of complex PTSD. Evolutionary perspectives have considerable potential to bridge and integrate neurobiology and the social sciences with respect to traumatic stress responses. PMID- 17464729 TI - Prodromal symptoms to relapse in bipolar disorder. AB - In a cyclical and recurring illness such as bipolar disorder, prodrome detection is of vital importance. This paper describes manic and depressive prodromal symptoms to relapse, methods used in their detection, problems inherent in their assessment, and patients' coping strategies. A review of the literature on the issue was performed using MEDLINE and EMBASE databases (1965-May 2006). 'Bipolar disorder', 'prodromes', 'early symptoms', 'coping', 'manic' and 'depression' were entered as key words. A hand search was conducted simultaneously and the references of the articles found were used to locate additional articles. The most common depressive prodromes are mood changes, psychomotor symptoms and increased anxiety; the most frequent manic prodromes are sleep disturbances, psychotic symptoms and mood changes. The manic prodromes also last longer. Certain psychological interventions, both at the individual and psychoeducational group level, have proven effective, especially in preventing manic episodes. Bipolar patients are highly capable of detecting prodromal symptoms to relapse, although they do find the depressive ones harder to identify. Learning detection, coping strategies and idiosyncratic prodromes are elements that should be incorporated into daily clinical practice with bipolar patients. PMID- 17464730 TI - CAOS contribution to understanding cultural/ethnic differences in the prevalence of bipolar affective disorder in New Zealand. AB - OBJECTIVE: To examine whether data on the clinical profiles of psychiatric service users can inform the claim that Maori in the New Zealand community have an increased rate of bipolar disorder. METHOD: The standardized recordings of a variety of clinical phenomena previously collected in a New Zealand study of psychiatric service users was extracted for those persons who were diagnosed with a bipolar mental disorder. The individual clinical phenomena were then compared by ethnicity. RESULTS: The most dramatic result was the increased recording of high levels of overactivity or disruptive aggressive behaviour in Maori compared with European in psychiatric service users, despite which, the rates of the disorder did not significantly differ by ethnicity. CONCLUSION: In the community survey lay raters were deciding on the presence or absence of phenomena including hyperactivity without the contextual knowledge that clinicians use. It is possible that the Composite International Diagnostic Interview decision tree was being fed an excessive amount of that criterion. PMID- 17464731 TI - Comparison of two anticonvulsants in a randomized, single-blind treatment of hypomanic symptoms in patients with bipolar disorder. AB - OBJECTIVE: Oxcarbazepine was compared to divalproex to assess clinical effectiveness of a proven agent, divalproex, against a newer, less studied agent, oxcarbazepine, in the treatment of hypomania. METHOD: Thirty patients with bipolar disorder, currently hypomanic, were randomized to receive oxcarbazepine or divalproex as add-on or monotherapy for 8 weeks. A rater blind to treatment assignment performed all symptom ratings. Hypomania and depression were rated using the Young Mania Rating Scale (YMRS) and the Inventory of Depressive Symptoms-Clinician Version (IDS-C). Random regression models were used to assess clinical symptom scores. RESULTS: There were no significant differences of YMRS or IDS-C scores between groups. Mean YMRS scores at baseline were 22.07+/-5.86 and 20.53+/-6.02 for the oxcarbazepine and the divalproex groups, respectively. Mean percent reduction from baseline to week 8 for the YMRS was 63.8% and 79.0% for oxcarbazepine and divalproex groups, respectively. Mean percent reduction from baseline to week 8 for the IDS-C was 48.7% versus 19.7% for oxcarbazepine and divalproex groups, respectively. Significant antimanic efficacy was noted for each medication. Both medications were generally well tolerated. CONCLUSION: In this pilot study, oxcarbazepine was as effective as divalproex in the treatment of hypomania. Further controlled trials are warranted. PMID- 17464732 TI - Taxometric investigation of depression: evidence of consistent latent structure across clinical and community samples. AB - OBJECTIVE: Past taxometric studies of depression have yielded equivocal results. Diversity of sample type may provide one explanation for this. The aim of the present study was to examine the latent structure of depression across clinical and community samples using exactly the same taxometric procedures involving exactly the same indicators of depression. METHOD: Two taxometric procedures, MAXEIG (maximum eigenvalue) and MAMBAC (mean above minus mean below a cut), were carried out on a clinical sample of 960 outpatients with mood and anxiety disorders. Simulated categorical and dimensional data sets as well as other consistency tests aided in the interpretation of the research data. Results were compared to a prior taxometric analysis in a community sample. RESULTS: The results of the current taxometric analyses were consistent with a dimensional latent structure and were compatible with the findings from identical analyses in a community sample. CONCLUSIONS: The findings of the current study highlight the importance of identifying factors that may contribute to, and explain, differences in the identified latent structure of depression. PMID- 17464733 TI - Venlafaxine hyponatraemia: incidence, mechanism and management. AB - OBJECTIVE: This prospective study was performed on patients aged >65 years commencing therapy with venlafaxine, in order to determine the incidence of hyponatraemia induced by the drug, to investigate the underlying pathophysiological mechanisms, and to evaluate a simple approach to management of this condition. METHOD: All patients aged >65 years seen by one author (MR) from all referral sources were entered into the study. Baseline biochemical tests were ordered, and if hyponatraemia developed (plasma Na <130 mmol L(-1)) additional tests were performed to ascertain the mechanism, while the patient continued on venlafaxine and fluid restriction was instituted. RESULTS: A total of 58 patients were seen, of whom 10 developed hyponatraemia, giving an incidence of 17.2%. Of these 10 patients, five were excluded from prolonged observation because of either severe medical illness, side-effects from the antidepressant or being lost to follow up. When hyponatraemia developed, it invariably did so within a few days of starting venlafaxine, and was associated with non-suppression of antidiuretic hormone in the face of a low serum osmolality. Fluid restriction (800 mL day(-1)) was effective in raising the plasma sodium to the normal range within 2 weeks, after which the fluid restriction could be relaxed without relapse occurring. These patients remained well for the follow-up period of up to 6 months. CONCLUSIONS: Patients >65 years of age should have their electrolytes measured 3-5 days after starting venlafaxine therapy. If hyponatraemia develops, it can be managed with modest fluid restriction without discontinuing drug treatment, subject to close continued clinical observation and biochemical monitoring. PMID- 17464734 TI - Assessing the impact of media guidelines for reporting on suicides in Austria: interrupted time series analysis. AB - OBJECTIVE: Media guidelines for reporting on suicides are a widely used means of preventing imitative suicides, but scientific accounts of their impact on suicide numbers are sparse. This report provides an evaluation of the Austrian guidelines that were introduced in 1987 as a natural experiment. METHODS: The impact of the guidelines was tested by applying an autoregressive integrated moving average (ARIMA) model and a linear regression model. In addition to a nationwide evaluation, Austria was divided into three areas according to regional differences in coverage rates of the collaborating newspapers and the impact of the intervention was tested for each area separately. Main outcome measures were the overall annual suicide numbers, and the numbers of Viennese subway suicides that were exceptionally newsworthy for the mass media. In order to test intermediate impacts, also quantitative and qualitative changes in media reporting after the introduction of the guidelines were analysed. RESULTS: There was some evidence of a nationwide impact of the guidelines, calculated as a significant reduction of 81 suicides (95% confidence interval: -149 to -13; t = 2.32, df = 54, p <0.024) annually. This effect was particularly due to a significant reduction in the area with the highest coverage rates of the collaborating newspapers. Viennese subway suicides showed a highly significant level shift (t = -4.44, df = 19, p <0.001) and a highly significant trend change (t = -4.20, df = 19, p <0.001) after the introduction of the guidelines. These effects corresponded to significant changes in the quality and quantity of media reporting. CONCLUSIONS: The present results clearly support the hypothesis that the media guidelines have had an impact on the quality of reporting as well as on suicidal behaviour in Austria, and stress the importance of collaborating with nationwide, but also with regional media to achieve efficacy. Further research is needed to provide an international insight into this public health issue. PMID- 17464735 TI - Mental health status of Aboriginal and non-Aboriginal Australian prisoners. AB - OBJECTIVE: To compare the mental health of Aboriginal and non-Aboriginal prisoners in New South Wales. METHODS: The sample consisted of a cross-sectional random sample of sentenced prisoners, and a consecutive sample of reception prisoners. The sample was drawn from 29 correctional centres (27 male, two female) across New South Wales. Overall, 1208 men (226 Aboriginal), and 262 women (51 Aboriginal) participated in the study. Mental illness was detected using the Composite International Diagnostic Interview (CIDI-A) and a number of other screening measures incorporated into the programme. RESULTS: No differences were detected in mental illness between Aboriginal and non-Aboriginal men, apart from depression, which was lower in the latter group. Aboriginal woman were more likely than non-Aboriginal women to screen positive for symptoms of psychosis in the prior 12 months and have a higher 1 month and 12 month prevalence of affective disorder; they also had higher psychological distress scores. Suicidal thoughts and attempts were the same in both groups. CONCLUSIONS: These findings confirm that the demand for mental health services in prisons is considerable, and that Aboriginal women are one of the most vulnerable groups. Services and programmes providing an alternative to incarceration are needed, as are culturally sensitive approaches to treatment. PMID- 17464736 TI - Problem alcohol use in young Australian adults. AB - OBJECTIVE: To examine the extent to which excessive drinking in young adults is associated with alcohol abuse and dependence. METHOD: Cross-sectional analyses were conducted using data from the eighth wave of the Victorian Adolescent Health Cohort Study, which comprised 1943 Victorians currently aged 24-25 years drawn from 44 secondary schools across the state in 1992. The main outcome measures of interest were short-term risk drinking status (based on daily alcohol consumption) and long-term risk drinking status (based on total weekly alcohol consumption). RESULTS: Two out of 5 participants drank at moderate to high risk levels for short-term harm. Yet, because young people tend to drink on only 1-2 days a week, fewer (22%) were at moderate to high risk for long-term harm. Although 20% of the participants met criteria for a diagnosis of alcohol abuse or dependence, most of those in the moderate- to high-risk drinking categories were not diagnosed with either alcohol disorder. CONCLUSION: Excessive alcohol use in one or two sessions a week appears to be common in young Australian adults. While short- and long-term risky drinking is more common in those with an alcohol use disorder, the majority of moderate- and high-risk drinking is done by those who do not meet criteria for an alcohol use disorder. PMID- 17464737 TI - Quality of life of Chinese schizophrenia outpatients in Hong Kong: relationship to sociodemographic factors and symptomatology. AB - OBJECTIVE: To explore the relationships between sociodemographic and clinical factors and quality of life (QOL) in a cohort of Chinese schizophrenia outpatients. METHOD: Two hundred subjects with a diagnosis of DSM-IV schizophrenia aged 18-60 years were randomly selected, and their sociodemographic and clinical characteristics including psychotic and depressive symptoms, extrapyramidal symptoms (EPS), and quality of life were assessed. Correlation and multiple regression analyses were used to evaluate the relationships of sociodemographic, clinical data and QOL. RESULTS: Compared to normative data obtained for the general population in Hong Kong, significantly lower scores in physical, psychological, and social QOL domains were found in the patient group. History of suicidal attempts and the presence of positive, negative, depressive, anxiety and EPS symptoms were all significantly correlated with QOL in schizophrenia patients. After controlling for the effects of variables that were significantly correlated with QOL in the correlation analysis, however, only depressive symptoms were still significantly correlated with each QOL domain. Multiple regression analysis showed that depressive symptoms predicted all QOL domains, while positive symptoms predicted overall and physical QOL domains. CONCLUSIONS: Chinese outpatients with schizophrenia had poorer QOL than the general population. In this patient population, QOL was more strongly related to the severity of depressive symptoms and was independent of sociodemographic factors. PMID- 17464738 TI - Testing the validity of the Recovery Assessment Scale using an Australian sample. AB - OBJECTIVE: Mental health services in Australia are increasingly becoming recovery orientated. However, there are varying meanings for recovery and few measures that specifically target recovery outcomes. The current study aimed to assess the construct and concurrent validity of a patient self-report measure, the Recovery Assessment Scale (RAS). METHOD: Participants were 168 individuals with severe and persistent psychiatric disability who were participants in the Australian Integrated Mental Health Initiative (AIMhi) project. They completed self-report recovery and other mental health measures and their case workers completed the Health of the Nation Outcome Scales. Exploratory and confirmatory factor analyses were carried out to examine the factor structure of the RAS. RESULTS: Exploratory factor analysis of the RAS produced five factors that were replicated using confirmatory techniques. Each factor has satisfactory internal reliability (Cronbach alpha range = 0.73-0.91). The factors displayed convergent validity with positive and significant correlations with other recovery measures. Concurrent validity was demonstrated with significant but lower correlations with symptoms and clinician-rated measures of psychiatric functioning. CONCLUSION: The factors of the RAS are consistent with the consumer literature on recovery. Correlations with other variable suggest that the RAS is measuring something different from traditional symptom or functional mental health measures. Further research is needed to clarify the extent to which the RAS is able to capture the range of recovery experiences that have been described by patients. PMID- 17464739 TI - Mental illness and physical activity: therapists' beliefs and practices. PMID- 17464740 TI - Atomoxetine-induced hyponatremia. PMID- 17464741 TI - Utility of systematic studies of the immune function in obsessive-compulsive disorder patients. PMID- 17464743 TI - Topiramate treatment of aggression in male borderline patients. PMID- 17464744 TI - Epicutaneous exposure to anticoagulant rodenticide warfarin modulates local skin activity in rats. AB - Dermatotoxic effects of epicutaneous application of a first generation anticoagulant, warfarin (WF), were examined in rats. Selected parameters of skin activity were determined 24 hours following warfarin application by histomorphological and immunohistochemical analysis and by assessing some aspects of immunomodulatory potential of warfarin in skin. Increased number of mast cells, with degranulation at higher doses of warfarin was noted in warfarin treated skin. Mast cell presence coincided with changes in blood vessels and fibroblast appearance suggesting mast cell activity in warfarin treated skin. Signs of nuclear hypertrophy and anysonucleosys were noted by analysis of PCNA(+) cells in epidermis following warfarin application. Histomorphological changes were accompanied by immunemodulating activity in warfarin treated skin. This was judged by slightly increased numbers of CD3(+) cells in epidermis and superficial dermis and by production of organ cultured full thickness skin explants of factors with costimulatory activity in T-cell activation/proliferation assay. Presented data demonstrates the potential of warfarin to modulate local skin activity in rats. PMID- 17464745 TI - Moisturizing lotions can increase transdermal absorption of the herbicide 2,4 dichlorophenoxacetic acid across hairless mouse skin. AB - Moisturizing lotions can be an effective treatment for occupationally induced dry skin. These compounds are designed to be hygroscopic and retain water to keep the stratum corneum hydrated, while at the same time enhancing the horny layer to prevent increases in transepidermal water loss (TEWL). Skin hydration levels, however, are known to influence barrier properties. The purpose of this work was to compare skin moisture levels induced by four commercially available moisturizing lotions with their capacity as transdermal penetration enhancers using the herbicide 2,4-dichlorophenoxyacetic acid (2,4-D) as a model chemical. Further, the effect of moisturizing the skin after washing with sodium lauryl sulfate (SLS) on transdermal absorption was determined. Skin moisture levels were also measured noninvasively and were correlated to penetration enhancement. Hairless mouse skin was pretreated with commercially available moisturizing lotions either with or without SLS washing and in vitro permeability studies were performed with the herbicide 2,4-D. The data demonstrate that pretreatment with three of the four lotions tested increased the transdermal absorption of 2,4-D as evidenced by cumulative penetration or faster lag times (p < 0.05). Skin moisture levels correlated with the penetration enhancement capabilities of the lotion. Washing the skin with 5% SDS increased the transdermal absorption of 2,4-D (p < 0.05) and application of moisturizing lotions increased the absorption further. In summary moisturizing lotions may influence transdermal penetration of the skin, with the more effective moisturizers having a greater effect on 2,4-D absorption. PMID- 17464746 TI - Evaluation of the hen's egg test-chorioallantonic membrane (CAM) method in prediction of the eye irritation potential formulated personal wash products. AB - A large database has demonstrated a robust relationship between the chorioallantonic membrane (CAM) and the Draize rabbit eye tests. The precision, sensitivity, and direct correlation of the CAM with human eye evaluations were critically appraised in this work. A total of 24 studies, including 12 CAM assays and 12 human clinical studies were conducted. Both the CAM assay and the human tests demonstrated good interassay precision and a robust correlation between the CAM scores and the inflammatory responses of human eye bulbar conjunctiva, palpebral conjunctiva and scleral vessels. This provided sound evidence that the CAM assay could be a predictive screen for eye irritation assessment for personal wash products and a key element of the tiered scientific approach in supporting labeling claims such as "tear free". PMID- 17464747 TI - Cumulative skin irritation test of sanitary pads in sensitive skin and normal skin population. AB - OBJECTIVE: We performed a randomized, paired, double-blind trial involving 14 participants with self-declared normal skin and 15 participants with self declared sensitive skin to test the cutaneous compatibility of materials of low irritation potential for skin irritation (sanitary pads) elicited by repetitive topical application. METHODS: Two test products patches and two control substance patches were applied to the lateral aspect of the upper arm once daily under fully occlusive conditions for 4 days. The positive control patch was moistened with sodium lauryl sulphate (0.1% w/v) and the negative control patch was moistened with 0.9% sodium chloride, as were the two test material patches. The patches were removed, skin irritation (erythema) graded, and new patches applied after each 24-hour period. RESULTS: There was no significant difference in skin irritation from the test patches between the sensitive and normal skin groups. The two test patches and the negative control patch were significantly less irritating than the positive control in both test groups. Both test patches were comparable to the negative control patch in the normal and sensitive skin groups. CONCLUSION: The test materials produced similar degrees of skin irritation among both groups. Overall, there was no statistically significant difference in skin compatibility when the two feminine hygiene product test patches were worn by self-declared normal and sensitive skin groups. PMID- 17464748 TI - Absorption of lawsone through human skin. AB - Lawsone (2-hydroxy-1,4-naphthoquinone) is the principal color ingredient in henna, a color additive approved with limitations for coloring hair by the Food and Drug Administration (FDA) under 21 CFR 73.2190. In 2002, the scientific committee on cosmetics and non-food products (SCCNFP), now known as the scientific committee for consumer products (SCCP), evaluated the safety of lawsone as a coloring agent in hair dye products of the European Union (EU). The SCCNFP concluded that lawsone was mutagenic and not suitable for use as a hair coloring agent. As a result, studies were conducted to measure the extent of lawsone absorption through human skin. Lawsone skin absorption was determined from two hair coloring products and two shampoo products, all containing henna. [(14)C]-Lawsone (sp. act. 22.9 mCi/mmol) was added to each commercial product and the products were applied to dermatomed, nonviable human skin mounted in flow through diffusion cells perfused with a physiological buffer (HEPES-buffered Hanks' balanced salt solution, pH 7.4). Products remained on the skin for 5 minutes (shampoos) and 1 hour (hair color paste). For the henna hair paste products, 0.3 and 1.3% of the applied dose was absorbed into the receptor fluid in 24 hours while 2.2 and 4.0% remained in the skin. For both henna shampoo products, 0.3% of the applied dose was absorbed into the receptor fluid at 24 hours while 3.6 and 6.8% remained in the skin. For all products, most of the lawsone applied was washed from the surface of the skin (83-102%) at the end of the exposure period. Extended absorption studies were conducted for 72 hours to determine if skin levels of lawsone in the 24 hour studies might eventually be percutaneously absorbed. These studies determined that the majority of the lawsone remained in the skin with only a small but significant increase (for three out of four products) in receptor fluid values. Therefore, it appears that receptor fluid values would give a good estimate of lawsone absorption for an exposure estimate and that skin levels of lawsone need not be included. PMID- 17464750 TI - The International Society for Cellular Therapy annual meeting comes to Sydney in 2007. PMID- 17464749 TI - Cutaneous contact urticaria to pyrethrum-real?, common?, or not documented?: an evidence-based approach. AB - Reports suggest that pyrethrum, the insecticidally active extract from Chrysanthemum cinerariifolium, can induce Type I hypersensitivity reactions in humans. Using knowledge of pyrethrum chemistry and an evidence-based analysis of literature, whether current refined pyrethrum induces and/or elicits skin manifestations of contact urticaria was assessed. Current extraction and refinement techniques suggest that refined pyrethrum lacks the presence of significant, if any, proteins speculated to induce Type I hypersensitivity. Our interpretation suggests that no reports of Type I reactions presented in the literature fulfill the criteria for immunologic contact urticaria. Future patient testing with current commercial material should clarify its Type I immunologic potential, if any. PMID- 17464751 TI - Cell and gene therapy in Australia. AB - The expansion of human cells to produce cell therapeutic products for the treatment of disease is, with few exceptions, an experimental therapy. Because cell therapies involve a biological product, often with some genetic or other modification, they require extensive pre-clinical research and development. Cell therapy production processes and premises require licensing by the Therapeutic Goods Administration. In this review, timed to coincide with the international meetings of the ISCT and ISSCR in Australia, we describe some promising cell therapies currently under development. PMID- 17464752 TI - The use of T-cell directed cellular therapies in Australia. AB - The use of cellular therapy in Australia has been limited by the number of facilities deemed acceptable for the preparation of T cells for immunotherapy. In spite of this, a number of trials using cellular immunotherapy have been conducted and a number are underway or are in the planning stage, and selected examples of these are summarized in this review. PMID- 17464753 TI - DC research in Australia. AB - Australian researchers have contributed significantly to the understanding of DC biology and clinical application over the past 25 years. Active DC research programs are in place in all major centers, pursuing the key questions of DC phylogeny, physiology and clinical applicability. Pre-clinical and clinical research include the pathophysiology of DC in malignancy, autoimmunity, chronic viral infection, chronic renal failure and transplantation medicine. In addition, Australian laboratories have uncovered some of the subtle complexities of DC subsets, often utilizing novel investigational tools discovered in their laboratories. Above all, Australian DC research has benefited from the existence of a potent culture of active collaboration, which has led to key interactions between cellular immunologists, clinician scientists and clinical researchers. These collaborations have led to the emergence of DC research programs that extend from in vitro and animal models of DC biology through each step of clinical translation and into active clinical trials. PMID- 17464754 TI - Recent Australian experience with hemopoietic stem and progenitor cell expansion. AB - This review provides insight into two clinical trials conducted with ex vivo manipulated CD34+ cells. The first was an attempt to deliver a gene therapy for treatment of HIV and the second an attempt to improve rates of hemopoietic recovery with ex vivo generated myeloid cells. PMID- 17464755 TI - Dedifferentiation of cells: new approaches. AB - Reprogramming of a differentiated cell into a cell capable of giving rise to many different cell types, a pluripotent cell, which in turn could repopulate or repair sick or damaged tissue, would present beneficial applications in regenerative medicine. Somatic cell nuclear transfer may offer this possibility, but technical hurdles and ethical frameworks currently prevent application of this technology in several countries. As a result, alternative strategies to reprogramming cell fate are being developed. This review briefly addresses somatic cell nuclear transfer and focuses on recent non-nuclear transfer-based approaches for reprogramming somatic cells and enhancing their differentiation potential. These include the fusion of somatic cells with embryonic stem cells, the treatment of somatic cells with extract of pluripotent cells and the retroviral transduction of somatic cells to overexpress pluripotency genes. PMID- 17464756 TI - Transfer of PR1-specific T-cell clones from donor to recipient by stem cell transplantation and association with GvL activity. AB - BACKGROUND: The curative effects of GvL following transfer of donor-derived T cells during allogeneic stem cell transplantation (SCT) are well established. However, little is known about the nature, origin and kinetics of the anti leukemic T-cell responses involved. METHODS: We used quantitative real-time PCR (qRT-PCR) for interferongamma mRNA production (IFN-gamma) and PR1/HLA-A*0201 tetramer staining to detect PR1-specific CD8+ T-cell activity in a donor and a patient with CML. Unbiased strand switch anchored RT-PCR was used to further characterize specific clones in PR1 sorted CD8+ T-cell populations. RESULTS: We identified PR1-specific CD8(+) T-cell clones from a donor pre-transplant, and demonstrated their transfer in the recipient's blood post-SCT using molecular tracking of Ag-specific T-cell receptors. PR1-specific CD8(+) T-cell populations were polyclonal, with a range of functional avidities for cognate Ag, and displayed predominantly effector memory phenotype early post-SCT, suggesting active stimulation in vivo. Expansion of these PR1-specific CD8(+) T-cell clones in the recipient was followed by complete remission of CML. DISCUSSION: This report represents the first direct demonstration that PR1-specific CD8(+) T-cell clones can be transferred during SCT, and supports the feasibility of pre transplant vaccination strategies that aim to boost the number of anti-leukemic T cells in the graft. PMID- 17464757 TI - Mesenchymal stromal cell-like characteristics of corneal keratocytes. AB - BACKGROUND: The unique potential of mesenchymal stromal cells (MSC) has generated much research interest recently, particularly in exploring the regenerative nature of these cells. Previously, MSC were thought to be found only in the BM. However, further studies have shown that MSC can also be isolated from umbilical cord blood, adipose tissue and amniotic fluid. In this study, we explored the possibility of MSC residing in the cornea. METHODS: Human cornea tissues were chopped to fine pieces and cultured in DMEM supplemented with 10% FBS. After a few days, the crude pieces of cornea were removed. Isolated keratocytes that were adherent to tissue culture flasks were grown until confluency before being passaged further. The immunophenotype was evaluated by flow cytometry. Assays were performed to differentiate cultured cells into adipocytes and osteocytes. RESULTS: Isolated corneal keratocytes exhibited a fibroblastoid morphology and expressed CD13, CD29, CD44, CD56, CD73, CD90, CD105 and CD133, but were negative for HLA-DR, CD34, CD117 and CD45. These properties are similar to those of BM-MSC (BM-MSC). In addition, corneal keratocytes were able to differentiate into adipocytes and osteocytes. DISCUSSION: Our results indicate that corneal keratocytes have MSC-like properties similar to those of BM-MSC. This study opens up the possibility of using BM-MSC in corneal tissue engineering and regeneration. Furthermore, discarded corneal tissue can also be used to generate MSC for tissue engineering purposes. PMID- 17464758 TI - Simultaneous isolation of human BM hematopoietic, endothelial and mesenchymal progenitor cells by flow sorting based on aldehyde dehydrogenase activity: implications for cell therapy. AB - BACKGROUND: ALDH(br) cells express high aldehyde dehydrogenase (ALDH) activity and have progenitor cell activity in several contexts. We characterized human BM ALDH(br) cells to determine whether cell sorting based on ALDH activity isolates potentially useful populations for cell therapy. METHOD: We measured the expression of ALDH and cell-surface Ag by flow cytometry and compared the ability of sorted ALDH(br), and BM populations remaining after ALDH(br) cells were removed (ALDH(dim) populations), to develop into several cell lineages in culture. RESULTS: The ALDH(br) population comprised 1.2+/-0.8% (mean+/-SD, n=30) nucleated cells and was enriched in cells expressing CD34, CD117, CD105, CD127, CD133 and CD166, and in primitive CD34(+) CD38(-) and CD34(+) CD133(+) progenitors. Most of the CD34(+) and CD133(+) cells were ALDH(dim). ALDH(br) populations had 144-fold more hematopoietic colony-forming activity than ALDH(dim) cells and included all megakaryocyte progenitors. ALDH(br) populations readily established endothelial cell monolayers in cultures. Cells generating endothelial colonies in 7 days were 435-fold more frequent in ALDH(br) than ALDH(dim) populations. CFU-F were 9.5-fold more frequent in ALDH(br) than ALDH(dim) cells, and ALDH(br) cells gave rise to multipotential mesenchymal cell cultures that could be driven to develop into adipocytes, osteoblasts and chondrocytes. DISCUSSION: Hematopoietic, endothelial and mesenchymal progenitor cells can be isolated simultaneously from human BM by cell sorting based on ALDH activity. BM ALDH(br) populations may be useful in several cell therapy applications. PMID- 17464759 TI - An internal positive control for the enumeration of CD45(+) and CD34(+) cells by flow cytometry allows monitoring of reagent and operator performance. AB - BACKGROUND: Three-color flow cytometry assays are used to determine CD34(+) cell doses prior to stem cell transplantation. These assays require high-quality reagents that are dispensed accurately to ensure reproducible results. We have developed a flow cytometry assay for CD34(+) cells with an integral positive control (KG1a cells) for monitoring reagent and operator performance. METHODS: The method was validated using samples from 127 allogeneic donations (42 BM, 85 PBSC) from healthy donors and 195 autologous donations (46 BM, 149 PBSC) from patients in remission from hematologic malignancies. The mean, SD and range of CD45(+) and CD34(+)cell counts were determined for each donation type. An internal control was used to assess performance of reagents and operators by comparison with a predetermined target value and an experienced operator. RESULTS: Replicate studies showed the method to be accurate and precise, with KG1a cells at 97.7+/-3.9% of the target value and a CV of 4.0%. In routine use over 322 samples, the accuracy was 91.7+/-17.7% of the target value, with a CV of 19.3%. Investigations into the cause of the reduced precision showed that reagents performed consistently well but operator performance was variable, with two of six operators significantly under-dispensing KG1a cells. DISCUSSION: This study validates our three-color flow cytometry assay and demonstrates that KG1a cells may be used to monitor test performance in the routine working environment. In addition to monitoring performance within a single laboratory, its wider use in multicenter studies may be helpful regarding standardization of methods. PMID- 17464760 TI - Transplantation of embryonic stem cell-derived cardiomyocytes improves cardiac function in infarcted rat hearts. AB - BACKGROUND: Post-infarct congestive heart failure is one of the leading causes of morbidity and mortality in industrialized countries. The main purpose of this study was to investigate whether transplantation of embryonic stem cell-derived cardiomyocytes (ESCM) directly into the infarcted myocardium could improve cardiac function in rats. METHODS: Cell culture medium with or without ESCM was injected into the borders of cardiac scar tissue 1 week after experimental infarction. Cardiac performance was evaluated 4 weeks later by means of echocardiography after ESCM (n=16) or medium (n=12) injection. RESULTS: ESCM implantation significantly improved fractional shortening (31.5+/-3. 8%) compared with medium-treated hearts (21.3+/-5.2%; P<0.05) and preserved left ventricular structure. Co-localization of 4',6-diamidino-2-phenylindole-labeled nuclei of transplanted cells with cardiomyocyte markers for cardiac troponin T and connexin 43, as detected by immunofluorescent microscopy, indicated the regeneration of damaged myocardium and the formation of gap junctions between grafted and host cells. However, intra-myocardial teratomas were observed in the hearts of two of the 16 grafted animals, at the fourth week after ESCM transplantation. DISCUSSION: Our results suggest that, although ESCM implantation can improve the function of infarcted myocardium, strategies to prevent tumorigenesis should be developed. PMID- 17464761 TI - A comparison of CFU-GM, BFU-E and endothelial progenitor cells using ex vivo expansion of selected cord blood CD133(+) and CD34(+) cells. AB - BACKGROUND: CD133 is a newly developed hematopoietic stem cell marker but little is known about its function. Whether CD133(+) cell selection provides any advantage over CD34(+) selection for hematopoietic stem cell isolation and transplantation is unclear. The present study compared colony formation and endothelial cell differentiation of these two cell types from umbilical cord blood (UCB). METHODS: Mononuclear cells from the same UCB samples were used for both CD133(+) and CD34(+) cell selection. Cells with 97.1% purity were incubated in semi-solid culture medium containing stem cell growth factor (SCGF) and G-CSF or erythropoietin (EPO). Purified cells were also cultured in M199 containing vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), and insulin-like growth factor-1 (IGF-1). RESULTS: CD34(+) and CD133(+) cells produced similar numbers of CFU-GM colonies (median 43.25 and 30.5, respectively; P>0.2). However, a greater than four-fold difference in BFU-E colony formation was observed from CD34(+) cells compared with CD133(+) cells (median 35 and 8, respectively; P<0.04). CD34(+) cells gave rise to endothelial-like cells when stimulated with VEGF, bFGF and IGF-1. CD133(+) cells were unable produce this cell type under the same conditions. DISCUSSION: CD133(+) cells produced smaller BFU-E colonies and were unable to differentiate into mature endothelial cells. CD34(+) cells contained endothelial progenitors that could differentiate into mature cells of this lineage. Based on these data, it appears that CD133 offers no distinct advantage over CD34 as a selective marker for immunoaffinity-based isolation of hematopoietic stem cells and endothelial progenitor cells. PMID- 17464762 TI - Subsets of mesenchymal stromal cells. PMID- 17464763 TI - Formyl peptide receptor expression in birds. AB - N-formyl-methionyl-leucyl-phenylalanine (fMLP) is a major chemotactic factor produced by Escherichia coli and other Gram-negative bacteria. The prototypal human fMLP receptor 1 (FPR1) was cloned in 1990 from a differentiated HL-60 myeloid leukemia cell cDNA library. In transfected cells, FPR1 binds fMLP with high affinity and is activated by picomolar to low nanomolar concentrations of fMLP in chemotaxis and calcium ion mobilization assays. Two additional human genes, designated FPR-like 1 (FPRL1) and FPR-like 2 (FPRL2), were later isolated by low-stringency hybridization using FPR1 cDNA as a probe, and these were shown to cluster with FPR1 on chromosome 19q13.3. In avian models the fMLP effects and the possible expression of FPRs have been poorly investigated. In this study we demonstrated that stimulation with fMLP of cultured cells isolated from the 10 day chick embryo brain causes superoxide anion and nitric oxide release and protein phosphorylation at serine, threonine, and tyrosine residues. These effects were abrogated by pretreatment with pertussis toxin, suggesting the involvement of a G-protein-coupled receptor (GPCR). Although specific N-formyl peptide receptors have so far been demonstrated only in mammals, a specific polyclonal antihuman-FPR1 antibody proved to bind to the membrane of both neurons and glial cells isolated from the chick brain. Immunoblot analysis revealed a single band corresponding to 60 kDa ca. A BLAST search and aa sequence alignments demonstrated that a number of avian 7-transmembrane (7TM) GPCRs share some homologies with the human FPR1. Furthermore, the CXCR4 ligand, SDF-1alpha, seems to compete with the antihuman-FPR1 polyclonal antibody used in our experiments. We thus advance the hypothesis that in birds one (or more) of the expressed 7TM GPCRs, most probably chemokine receptors belonging to the CXCR4 subfamily, also may act as fMLP receptors. PMID- 17464764 TI - Effect of intrauterine exposure of murine fetus to cyclophosphamide on development of thymus. AB - The objective of this study was to demonstrate thymic alterations produced by cyclophosphamide intervention during intrauterine life of murine fetus. Cyclophosphamide (CP) was administered to pregnant mice on day 11 of gestation in a single dose of 10 mg/kg body weight. Fetuses were dissected out on day 19 and studied for various effects on thymus. Thymus of fetuses exposed to cyclophosphamide showed thymic atrophy with retardation of thymic size and a remarkable shrinkage in lobular morphology. Histological studies showed a massive depletion of thymic cortex. Study of thymocytes revealed an increase in apoptotic cell count and percent DNA fragmentation along with a decrease in proliferation. Thymocytes obtained from fetuses of CP-treated mice showed a higher expression of caspase-activated DNase (CAD) indicating that the CP-dependent induction of apoptosis in thymocytes involved caspase pathway. The results of the present study may help in understanding the mechanism of the teratogenic effect of cyclophosphamide on thymus. PMID- 17464765 TI - Separation methods of T cells, natural killer, and dendritic cells from peripheral blood of cancer patients using interleukin-2 and functional analysis of natural killer cells after separation. AB - We aimed to induce three different immune cell subsets from a single blood sample from cancer patients to target different biological characters of cancer cells. In the presence of 6000 IU/ml IL-2, natural killer (NK) cells adhere to plastic. By using this ability, we could separate dendritic cells, T cells, and NK cells from peripheral blood mononuclear cells. The cultured NK cells demonstrated higher nonspecific cytotoxicity against tumor cell lines than did the T cells. Furthermore, adherent NK cells demonstrated higher cytotoxicity than nonadherent NK cells, although there was no difference between adherent and nonadherent NK cells in natural cytotoxicity receptors (NKp30, NKp44, NKp46) and NKG2D expression. With these results, we confirmed that we could induce dendritic cell, T cell, and higher cytotoxic NK cells from a single blood draw, and this methodology facilitates to the use of these cells for clinical grade conditions. PMID- 17464766 TI - Therapeutic approach by a novel designed anti-inflammatory drug, M2000, in experimental immune complex glomerulonephritis. AB - The therapeutic efficacy of novel designed nonsteroidal anti-inflammatory drug, M2000 (beta- D- mannuronic acid) on experimental immune complex glomerulonephritis was evaluated. Bovine serum albumin (BSA) nephritis was induced in rats by a subcutaneous immunization and daily intravenous administration of BSA. M2000 solution (30 mg/kg) was administered intraperitoneally at regular 48-hr intervals for 4 weeks. Onset of treatment was day 56. Urinary protein was measured weekly and serum anti-BSA antibody was assessed by ELISA method at different intervals. Animals were killed on day 84 and blood samples and kidney specimens were obtained. Serum (creatinine, blood urea nitrogen, cholesterol, and triglyceride) and urine (protein, urea, and creatinine) determinants were measured at the time of sacrifice. Kidney specimens were processed for light and immunofluorescent microscopic examination. The fibrosarcoma cell line was used for assaying tolerability and matrix metalloproteinase type 2 (MMP-2) activity. MMP-2 activity was assessed using zymography. Our data showed that M2000 therapy could significantly reduce the urinary protein excretion in treated rats versus non-treated controls. Anti-BSA antibody titer was lower in treated rats than in controls at the 12th experimental week. Polymorphonuclear neutrophil leukocytes infiltration and glomerular immune complex deposition were less intense in treated rats than in controls. Cytotoxicity analysis of M2000 showed a much higher tolerability compared with other tested drugs (diclofenac, piroxicam and dexamethasone). The inhibitory effect of M2000 in MMP-2 activity was significantly greater than that of dexsamethasone and of piroxicam at a concentration of 200 microg/ml. Moreover, the toxicological study revealed that M2000 had no influence on serum (BUN, creatinine, triglyceride and cholesterol) determinants, urinary protein excretion and glomerular histology in healthy group receiving drug. CONCLUSIONS: These findings suggest that treatment with M2000 can reduce proteinuria, diminish antibody production, and suppress the progression of disease in a rat model of immune complex glomerulonephritis. PMID- 17464767 TI - Role of interleukin-6 in toll-like receptor 4 and 2 expressions induced by lipopolysaccharide in the lung. AB - Our previous study demonstrated that interleukin (IL)-6 is protective against hemorrhagic lung inflammation induced by lipopolysaccharide (LPS) in mice, at least partly, by inhibition of the enhanced expressions of proinflammatory cytokines. The present study elucidated the role of IL-6 in Toll like receptor (TLR) 4 and 2 expressions in the lung during inflammation induced by intraperitoneal administration of LPS (1 mg/kg) using IL-6 null (-/-) mice and wild type (WT) mice. The expressions of mRNA for both TLR4 and 2 in the lung were evaluated 72 hrs after intraperitoneal administration. LPS enhanced both TLR4 mRNA expression as compared with vehicle. However, the enhancement was significantly stronger in IL-6 (-/-) mice than in WT mice after LPS challenge. LPS significantly increased TLR2 mRNA expression only in IL-6 (-/-) mice. As well, in the presence of LPS, the expression was significantly greater in IL-6 ( /-) mice than in WT mice. These results suggest that the protective role of IL-6 against LPS-induced hemorrhagic lung inflammation might be explained, in part, by inhibition of the enhanced lung expressions of TLR4 and 2. PMID- 17464768 TI - In vitro elicitation of intestinal immune responses in teleost fish: evidence for a type IV hypersensitivity reaction in rainbow trout. AB - In fish the gut immune system has been the subject of few investigations until now. Here, we provide novel morphological and immunological data on the gut isolated from rainbow trout Salmo gairdneri. The pyloric (P) and terminal (T) segments of trout gut, when morphologically examined, evidenced lymphocytes and macrophages (MO) loosely dispersed in the intestinal mucosa and in the lamina propria in the absence of typical Peyer's patches-like structures. Furthermore, incubation of P and T sections with Candida albicans (Ca) and functional analysis of supernatants generated some interesting results. In fact, active supernatants, when compared with controls, exhibited cytokine-like activities attributable to the presence of interferon (IFN)-gamma and migration inhibiting factor (MIF), respectively. In particular, IFN-gamma-like activity gave rise to an enhancement of Ca phagocytosis by MO, whereas MIF inhibited MO migration in agarose. Taken together, these in vitro data suggest that the gut-associated lymphoreticular tissue in fish possesses the appropriate armamentarium to mount a type IV hypersensitivity response when challenged by microbial antigens. PMID- 17464769 TI - Activation of cytotoxic T lymphocyte responses and attenuation of tumor growth in vivo by Andrographis paniculata extract and andrographolide. AB - The stimulatory effect of Andrographis paniculata extract and andrographolide on cytotoxic T lymphocyte (CTL) production was determined in BALB/c mice by Winn's neutralization assay using CTL sensitive EL4 thymoma cells as target cell. Extract and andrographolide showed a significant increase in CTL production in both the in vivo and in vitro models. The survival time of EL4 cells alone in animals was only 27.1 days and it was increased to 51.1 and 44.5 days in extract- and andrographolide treated animals with percentage increase in life span (%ILS) of 88.5 and 64.2, respectively. The survival rate of animals administered EL4 cells incubated with alloimmunized spleen cells (effector cells) from normal BALB/c mice was 35.8 (%ILS 32.1). When this group was treated with 10 doses of extract and andrographolide the life span was further increased to 52.1 days (%ILS 92.2 ) and 48.1 days (%ILS 77.4). Survival days of animal carrying EL4 cells incubated with alloimmunized spleen cells (effector cells) from extract and andrographolide treated animals were 55.5 and 50.3 days respectively. When these animals continued with extract and andrographolide treatment for 10 days their life spans were significantly increased to 62 and 53.8 days, respectively. The level of cytokines such as Interlevkin (IL)-2 and Interferon (IFN)-gamma also was enhanced in these animals when they were treated with extract and andrographolide. This study demonstrated that A. paniculata extract and andrographolide stimulate the CTL production through enhanced secretion of IL-2 and IFN-gamma by T cells and thereby inhibit the tumor growth. PMID- 17464770 TI - Ex vivo triggering of T-cell-mediated immune responses by autologous tumor cell vaccine in oral cancer patients. AB - We investigated immunomodulatory activity of autologous tumor cell vaccine from oral cancer patients ex vivo by lymphoproliferation assay and two color flow cytometry. Vaccine treatment lead to 10-fold higher proliferation of lymphocytes compared with the untreated controls. A significant increase in CD69(+) and HLA DR(+) T-cells was observed in vaccine pulsed cultures compared with untreated (p<0.0001) controls. The frequency of IFN-gamma and IL-2 expressing CD4(+)/CD8(+)T-cell subsets was significantly higher with a concomitant reduction in IL-4 and IL-10 expression in the vaccine-treated group (p<0.0001) compared with the untreated controls. Vaccine treatment further increased T-cell receptor Vbeta3, Vbeta5, and Vbeta8 usage. It seems that the autologous tumor cell vaccine triggers T-cell responses ex vivo, hence it may have a protective role in oral cancer patients. PMID- 17464771 TI - Cyclosporin a and tacrolimus (FK506) differentially alter T-cell receptor expression in vivo. AB - Cyclosporin A (CSA) and tacrolimus (FK506) are two common immunosuppressive agents used post blood and marrow transplantation. Despite similarity in their accepted modes of action, we observed polarized effects of CSA and FK506 on the in vivo human T cell repertoire. To determine the possible mechanism for this difference, the effects of CSA and FK506 on cell viability, cell proliferation, interleukin-2 production, and calcineurin inhibition were determined in vitro. Our data suggest that a secondary mechanism of action exists for the different T cell repertoire induced by exposure to CSA and FK506. PMID- 17464772 TI - Enhancement of 1,25-dihydroxyvitamin D3- and all-trans retinoic acid-induced differentiation of human leukemia HL-60 cells by Phyllostachys nigra var. henonis. AB - Human myeloid leukemia HL-60 cells are differentiated into monocytic or granulocytic lineage when treated with 1,25-dihydroxyvitamin D(3) [1,25 (OH)(2)D(3)] or all-trans retinoic acid (RA), respectively. In this study, the effect of acetone fraction prepared from bamboo leaf on cell differentiation was investigated in a HL-60 cell culture system. Treatment of HL-60 cells with 50-400 microg/ml acetone fraction of bamboo leaf for 72 hr inhibited cell proliferation and induced a little increase in cell differentiation, as demonstrated by the MTT and nitroblue tetrazolium reduction assay. Interestingly, synergistic induction of HL-60 cell differentiation was observed when the acetone fraction of bamboo leaf was combined with either 5 nM 1,25-(OH)(2)D(3) or 50 nM all-trans RA. Flow cytometric analysis indicated that combinations of 1,25-(OH)(2)D(3) and the acetone fraction of bamboo leaf stimulated differentiation predominantly to monocytes, whereas combinations of all-trans RA and the acetone fraction of bamboo leaf stimulated differentiation predominantly to granulocytes. These results suggest that the acetone fraction of bamboo leaf enhanced leukemia cell differentiation and suggest a possibility of bamboo in the treatment of leukemia. PMID- 17464773 TI - Inhibition of interleukin-12 production in mouse macrophages via suppression of nuclear factor-kappaB binding activity by Phyllostachys nigra var. henonis. AB - Pharmacological inhibition of interleukin-12 (IL-12) production may be a therapeutic strategy for preventing development and progression of disease in experimental models of autoimmunity. The acetone fraction prepared from bamboo, Phyllostachys nigra var. henonis, potently inhibited the Lipo polysaccharide (LPS)-induced IL-12 production from RAW264.7 monocytic cell-line in a dose dependent manner. The repressive effect mapped to a region in the IL-12 gene promoter containing a binding site for NF-kappaB. Furthermore, activation of macrophages by LPS resulted in markedly enhanced binding activity to the NF kappaB site, which significantly decreased upon addition of the acetone fraction of Phyllostachys nigra var. henonis. This indicated that the acetone fraction inhibited IL-12 production in LPS-activated macrophages via inhibition of NF kappaB binding activity. PMID- 17464774 TI - Antigenically activated avian erythrocytes release cytokine-like factors: a conserved phylogenetic function discovered in fish. AB - Fish erythrocytes are endowed with the ability to produce cytokine like factors when stimulated with Candida albicans (Ca). To evaluate whether similar activities are still conserved in bird erythrocytes (BE), a morphological, cytochemical and immunological evaluation was conducted on peripheral cells in chickens (Gallus gallus). BE form rosettes with monocytes (Mo)-macrophages (MO), and Mo-MO according to cytochemical analysis to maintain phagocytic functions across the evolution. Finally, Ca-activated BE release in the supernatants cytokine like-factors that enhance Mo-MO phagocytosis (interferon-gamma-like activity) and inhibit Mo-MO migration in agarose (migration inhibitory factor activity). In conclusion, bird erythrocytes, as nonimmune cells, are able to participate in the immune response contributing to the host defence. PMID- 17464775 TI - Smoking cessation after acupuncture treatment. AB - Acupuncture is applied, especially in treatment of pain, hemiplegia, obesity, and psychological illnesses including addiction. Recently, ear and body acupuncture have been frequently used in the treatment of smoking. An increase in levels of endorphin, enkephalin, epinephrine, norepinephrine, serotonin, and dopamine in the central nervous system and plasma has been reported as the most important mechanism of acupuncture. That is, acupuncture application may increase the levels of endorphin, enkephalin, epinephrine, norepinephrine, serotonin, and dopamine in the central nervous system and plasma. The authors think that acupuncture application provides the patients with deterioration in the taste of smoking, decrease in desire of smoking, and the obstruction of psychological symptoms that appear as a result of smoking cessation. Because of these effects it is presumed that acupuncture application may be used as an important method for smoking cessation treatment. PMID- 17464776 TI - Electroacupuncture treatment of obesity with psychological symptoms. AB - The aim was to study the effect of placebo EA, electroacupuncture (EA), and diet on obesity and accompanying psychological symptoms. One hundred and sixty-five volunteer women participated in the study. There were three groups: (i) Placebo EA, (ii) EA, and (iii) diet restriction group. EA was performed by using three ear and six body points. There was a 4.8% reduction in weight of patients with EA application, whereas patients with a diet restriction and placebo EA had a 2.5% and 2.7% weight reduction, respectively. There were significant decreases in phobia, anger, anxiety, obsession, paranoid symptoms, and depression in the EA groups compared to those of the placebo EA and diet groups. It was suggested that electroacupuncture may be an effective therapy for obesity including the psychological signs and symptoms in women. PMID- 17464777 TI - Increased temperature at acupuncture points induced by weight reduction in obese patients: a preliminary study. AB - This study investigated the temperature changes at acupuncture points resulting from a weight reduction program. A standard method using a digital infrared thermographic imaging device was employed to monitor the temperature at 20 acupuncture points in 32 participants treated for obesity at an Oriental obesity clinic. There were significant temperature increases at all of the acupuncture points except for LI4 (Hegu) and TE5 (Waiguan). The mean total temperature change at the acupuncture points was significantly correlated with the BMI change (r = .588, p < .001). The weight-reduction-induced increase in temperature at acupuncture points may reflect improvements in the circulation of energy flows. However, further study is necessary to understand fully the relationship between the temperature at acupuncture points and weight reduction, including the use of appropriate controls. PMID- 17464778 TI - A comparison of voluntary and involuntary measures of electromechanical delay. AB - Electromechanical delay (EMD) is a measurement used to assess the mechanical lag between muscle activation onset and force production. EMD measurements may be performed by voluntary or electrically evoked muscle activation. This study compared gastrocnemius EMD during voluntary and involuntary contractions and assessd the intrasession reliability of each set of measurements. Subjects were 15 volunteers (age 21 +/- 2 years, ht 171.8 +/- 10.0 cm, mass 76.1 +/- 13.4 kg). EMD measurements were recorded from the medial head of the gastrocnemius of each subject during voluntary and involuntary contractions. Order was counterbalanced between subjects. Subjects stood with the dominant leg on a force plate, the nondominant next to the force plate, and with their hands in contact with a bar in front of them. A supramaximal percutaneous stimulus was applied to the tibial nerve in the popliteal fossa for involuntary (electrically evoked) contractions. For voluntary contractions, subjects were instructed to rise up on the toes as quickly as possible. Four trials were collected for each condition with 30 s of rest between each. Repeated measures ANOVAs were performed for each condition to calculate an ICC (2,1). Means of the 4 trials for each condition were used to detect differences between groups. EMD was greater in the voluntary condition (22.8 +/- 8.2 ms) compared to the involuntary condition (9.7 +/- 3.1 ms; p < .001). Intrasession reliability for each condition was very strong (involuntary ICC (2,1) = .977; voluntary ICC (2,1) = .972). EMD measured during a single leg stance is much shorter when measured during an electrically evoked (voluntary) contraction. The difference in EMD between conditions is likely the result of differences in recruitment during the two types of contractions. Reliability within a measurement session was very strong for each of the conditions. PMID- 17464779 TI - Behavioral and psychophysiological characteristics of children of parents with social phobia: a pilot study. AB - Recent studies have noted a relation between the pattern of resting frontal EEG activity and individual differences in affective style in typically developing infants, children, and adults. The authors conducted a pilot study to investigate the pattern of frontal EEG activity during a resting condition (eyes-open, eyes closed) in a group of children who had one parent clinically diagnosed with social phobia (SP; n = 6) and in a group of typically developing children of similar age with healthy parents (n = 7). Patients with a primary DSM-IV diagnosis of SP with at least one biological child were recruited from the Anxiety Disorders Clinic at McMaster University Medical Centre. We found that children of parents clinically diagnosed with SP tended to exhibit higher overall resting frontal EEG activity compared with the children of healthy parents. This pattern of overall high EEG activity that is specific to the frontal region is similar to that observed in socially anxious profiles. Preliminary findings are discussed in terms of how overall resting frontal brain activation may be an early psychophysiological marker for placing children of parents with social phobia at risk for socioemotional problems before such problems emerge. PMID- 17464780 TI - Effects of interleukin-1beta on spinal cord nociceptive transmission in intact and propentofylline-treated rats. AB - To investigate the contribution of glial cells in the spinal cord nociceptive transmission, the effect of intrathecally administered interleukin-1beta (IL 1beta) was studied in rats treated with the glial cell inactivator propentofylline and submitted to a C-fiber-mediated reflex paradigm evoked by single and repetitive (wind-up) electric stimulation. Intrathecal IL-1beta did not modify the C reflex integrated activity in either group of animals, while producing increased wind-up in intact and decreased wind-up in propentofylline pre-treated rats. Results suggest that the excitatory effect of IL-1beta on spinal wind-up activity in healthy rats is produced by a glial mediator, whereas the inhibitory effect resulted from a direct effect of the cytokine on dorsal horn neurons. PMID- 17464781 TI - Coerulear activation by crh and its role in hypertension induced by prenatal malnutrition in the rat. AB - The effects of intracoerulear CRH and intraparaventricular prazosin on systolic pressure, diastolic pressure and heart rate were studied in prenatally malnourished hypertensive rats. At day 40 of life, (i) malnourished rats showed enhanced systolic pressure, heart rate, and plasma corticosterone; (ii) intracoerulear CRH increased systolic pressure and heart rate only in controls; (iii) intraparaventricular prazosin decreased systolic pressure and heart rate only in malnourished rats; (iv) in controls, prazosin did not prevent the stimulatory effect of CRH on the cardiovascular parameters; in malnourished rats, prazosin allowed CRH regain its stimulatory effects. Thus, coerulear activation by CRH would be involved in hypertension and tachycardia developed by prenatally malnourished animals. PMID- 17464782 TI - Positive affect modulates activity in the visual cortex to images of high calorie foods. AB - Activity within the visual cortex can be influenced by the emotional salience of a stimulus, but it is not clear whether such cortical activity is modulated by the affective status of the individual. This study used functional magnetic resonance imaging (fMRI) to examine the relationship between affect ratings on the Positive and Negative Affect Schedule and activity within the occipital cortex of 13 normal-weight women while viewing images of high calorie and low calorie foods. Regression analyses revealed that when participants viewed high calorie foods, Positive Affect correlated significantly with activity within the lingual gyrus and calcarine cortex, whereas Negative Affect was unrelated to visual cortex activity. In contrast, during presentations of low calorie foods, affect ratings, regardless of valence, were unrelated to occipital cortex activity. These findings suggest a mechanism whereby positive affective state may affect the early stages of sensory processing, possibly influencing subsequent perceptual experience of a stimulus. PMID- 17464783 TI - Effects of extract of ginkgo biloba on intracranial pressure, cerebral perfusion pressure, and cerebral blood flow in a rat model of subarachnoid haemorrhage. AB - The purpose of this study was to investigate in effect of extract of Ginkgo biloba (EGb) on cerebral blood perfusion in a subarachnoid haemorrhage (SAH) rat model. SAH lead to an increase in intracranial pressure and decrease in cranial perfusion pressure and regional cerebral blood flow in all groups. However, the intracranial pressure increases in EGb groups were less than that of the vehicle group (p < .01), whereas the reduction in cranial perfusion pressure and regional cerebral blood flow in the EGb group was less than that of the vehicle and SAH groups (p < .01). It was concluded that EGb attenuates the increase in intracranial pressure and reduction in cerebral blood perfusion after SAH. PMID- 17464784 TI - On reaction dynamics at dopamine synapses. AB - Dopamine neurons play a key role in normal and pathological cognitive processes as well as in the effects of certain drugs of addiction. Models of the synapses of such neurons include transporter mechanisms and reaction dynamics. This article focuses attention on the fundamental reaction that converts tyrosine to DOPA, which involves a cofactor. The Michaelis-Menten formula for the rate of an enzymatic reaction is modified by the presence of cofactors, which may be either essential or nonessential. In the essential case, the reaction rate is found to depend on the relative magnitudes of the concentrations of the primary enzyme and the cofactor. The case of a nonessential cofactor is more complex and it is shown for the first time how this leads to reaction rate formulas that depend explicitly on the concentrations of the enzyme and cofactor. The extended Michaelis-Menten formulas are applied to the aforementioned reaction with tyrosine hydroxylase as enzyme and biopterin as cofactor. The results are useful in constructing accurate models of dopamine synapses. PMID- 17464785 TI - Computational and noncomputational systems in brain and cognition: can one mask the other? AB - A theory is developed based on the premise that nonneural processes occur in the brain exemplified as spatial working memory, and is the seat of consciousness. Additionally, wave storage of spatial information, a Bose condensate to support the waves, and the location of wave storage are provided as illustrative, "existence proofs" that a coherent theory can be built along these lines in agreement with the data. The theory can be built argues on functional grounds that a nonneural spatial memory may serve a vital biological function. This article demonstrate how this same non-neural memory can bridge the explanatory gap to consciousness, in agreement with the facts. The article proposes a possible mechanism and location for the nonneural component. PMID- 17464786 TI - Performance on figure ground perception following stroke induced hemiplegia: a compairson of pre-and post-rehabilitation with the neurologically unimpaired. AB - This article compares performance times on the Figure Ground Perception (FGP) of the Sensory Integration and Praxis Tests (SIPT) in persons with stroke-induced hemiplegia and unimpaired older persons. Two hundred and twelve right dominant persons with hemiplegia on admission and discharge from comprehensive rehabilitation and 321 neurologically unimpaired persons in the community were evaluated on the FGP subtests of the SIPT. There were significant differences in performance between unimpaired persons and those with hemiplegia following a stroke. Among the persons with stroke-induced hemiplegia, performance on discharge was significantly better than on admission across all five FGP subtests. Age, sex, and severity of hemiplegia also affected performance on specific subtests; the influence of laterality was significant in persons with right hemispheric lesions; and persons with Wernicke's aphasia performed more poorly. Although further investigation is clearly needed, the association of increasing age (particularly males 75 years and over), severity of hemiplegia, and presence of aphasia on FGP performance suggest that these factors should be considered in the development of rehabilitation programs for stroke patients. PMID- 17464787 TI - At issue: anti-stigma-endeavours. PMID- 17464788 TI - Is antipsychotic medication stigmatizing for people with mental illness? AB - Antipsychotic medications are clearly identified as important in the treatment of individuals with schizophrenia and with bipolar disorder. However, negative societal reaction related to having a serious mental illness and the socially undesirable side effects associated with antipsychotic medication treatment may combine to worsen stigma associated with treatment for mental illness. Specific stigmatizing effects of antipsychotic therapy may be difficult to evaluate independently from factors such as symptoms, insight into illness and side effects. Attitudes towards antipsychotic medication may be positive in individuals who recognize therapeutic drug effects, however other individuals may view medications negatively due to a sense of stigma. Stigma among individuals with bipolar disorder in relation to treatment with antipsychotic medication has not been well addressed in the literature. An additional concern among individuals with bipolar disorder who receive antipsychotic medications may be the notion that antipsychotics are 'schizophrenia drugs', and thus an inappropriate treatment for their condition. Antipsychotic medications can be stigmatizing for patients with serious mental illness, however the roots of stigma are extensive, and efforts to minimize stigma can only be successful when addressed by the individual with illness, their families and loved ones, treatment providers and society at large. PMID- 17464789 TI - Discrimination in health care against people with mental illness. AB - This paper discusses factors associated with low rates of help-seeking and poorer quality of physical healthcare among people with mental illnesses. Evidence is reviewed on the associations between low rates of mental health literacy, negative attitudes towards people with mental illness, and reluctance to seek help by people who consider that they may have a mental disorder. People with mental illness often report encountering negative attitudes among mental health staff about their prognosis, associated in part with 'physician bias'. 'Diagnostic overshadowing' appears to be common in general health care settings, meaning the misattribution of physical illness signs and symptoms to concurrent mental disorders, leading to underdiagnosis and mistreatment of the physical conditions. PMID- 17464790 TI - The stigma of mental illness: anticipation and attitudes among patients with epileptic, dissociative or somatoform pain disorder. AB - The aim of this study was to survey the attitudes of 101 consecutive in- and out patients with epileptic, dissociative or somatoform pain disorders (mean age: 43 [+/-11] years; 58% female) from either the Department of Psychiatry or Neurology toward anticipated mental illness stigma. The patients were administered a modified 12-item version of Links Stigma Questionnaire. Nearly 60% of all 101 patients believe that "most people" would not allow a mental patient "to take care of their children", "most young women" would be "reluctant to date a man" who has been treated for a mental illness and "most employers would pass over" the application of a psychiatric patient in favour of another applicant. Fifty five percent of the respondents assume that "most people think less of a person who has been in a mental hospital" and over a half of all patients interviewed assert that the general population thinks that psychiatric patients are "less intelligent, less trustworthy and that their opinion is taken less seriously by others". Gender, age and education had no influence on the overall results. There is a high stigmatisation concerning psychiatry even in patients with epilepsy and somatoform/dissociative symptoms with psychiatric comorbidity. Fear of being stigmatized is more pronounced among somatoform pain patients as compared to patients suffering from epileptic or dissocative disorders, with particular reference to close personal relationships. PMID- 17464791 TI - Stigmatization, social distance and exclusion because of mental illness: the individual with mental illness as a 'stranger'. AB - The stigma attached to mental illness often leads to underestimation, underdiagnosis and undertreatment of mental disorders. A lack of knowledge of causes, symptoms and treatment options of mental disorders in the public and a lack of personal contact with affected individuals can result in prejudices and negative attitudes towards them-and subsequently in stigmatization and discrimination. Human beings suffering from mental illness often are recognized as 'strangers'. But, social-psychological and philosophical analysis of the recognition of 'strangeness' in mentally ill individuals as one of the main predictors for social distance towards them, shows, that the other person only remains a stranger if the available cognitive patterns of interpretation fail. Society must provide the people living in it with the interpretational patterns to take away the recognized strangeness from mentally ill persons and thus making him, or her, an accepted other member. PMID- 17464792 TI - Stigma and mental health professionals: a review of the evidence on an intricate relationship. AB - In the past decade, mental health professionals have initiated a number of national and international efforts against the stigma of mental illness. While largely successful in beating stigma and discrimination, these programmes have, in part, been criticized to be largely uninformed by the lived realities of people with mental illness and their families. Some critics claimed that anti stigma efforts led by mental health professionals were in fact a concealed attempt at de-stigmatizing psychiatry itself as a profession. This paper will attempt to throw light on the various ways in which mental health professionals are 'entangled' in anti-stigma activities. It will outline the complex relationships between stigma and the psychiatric profession, presenting evidence on how its members can simultaneously be stigmatizers, stigma recipients and powerful agents of de-stigmatization. In exploring the role of mental health professionals as targets of stigma, new findings will be presented on the role of stigma as a professional stressor in psychiatry. Conclusions will be drawn on how the pursuit of professional self-interest can be a legitimate goal of anti-stigma programmes. Further, ways in which acknowledging psychiatry's own agenda can contribute to both credibility and success of fighting stigma from within psychiatry will be discussed. PMID- 17464793 TI - Stigma towards people with mental illness in developing countries in Asia. AB - BACKGROUND: There is a wide range of literature on stigmatization and discrimination of people with mental illness. Most studies, however, derive from Western countries. This review aims at summarizing results from developing countries in Asia published between 1996-2006. METHOD: Medline search focusing on English-speaking literature. RESULTS: Comparable to Western countries, there is a widespread tendency to stigmatize and discriminate people with mental illness in Asia. People with mental illness are considered as dangerous and aggressive which in turn increases the social distance. The role of supernatural, religious and magical approaches to mental illness is prevailing. The pathway to care is often shaped by scepticism towards mental health services and the treatments offered. Stigma experienced from family members is pervasive. Moreover, social disapproval and devaluation of families with mentally ill individuals are an important concern. This holds true particularly with regards to marriage, marital separation and divorce. Psychic symptoms, unlike somatic symptoms, are construed as socially disadvantageous. Thus, somatisation of psychiatric disorders is widespread in Asia. The most urgent problem of mental health care in Asia is the lack of personal and financial resources. Thus, mental health professionals are mostly located in urban areas. This increases the barriers to seek help and contributes to the stigmatization of the mentally ill. The attitude of mental health professionals towards people with mental illness is often stigmatizing. CONCLUSION: This review revealed that the stigmatization of people with mental illness is widespread in Asia. The features of stigmatization-beliefs about causes of and attitudes towards mental illness, consequences for help-seeking have more commonalities than differences to Western countries. PMID- 17464794 TI - Olfactory jump reflex habituation in Drosophila and effects of classical conditioning mutations. AB - Habituation is a nonassociative learning mechanism, in which an initial response toward repeated stimuli gradually wanes. This is amongst the simplest and most widespread forms of behavioral plasticity. So far, neither the underlying molecular mechanisms nor the precise neural networks of habituation are well understood. We have developed a novel paradigm to quantify habituation of the olfactory jump reflex in Drosophila. We present data demonstrating several behavioral properties of this phenomenon, generally observed in other species. We also show that the dunce and rutabaga memory mutants behave abnormally in this assay, suggesting that this assay might be used in behavioral screens for new mutants with defects in this simpler form of behavioral plasticity. PMID- 17464795 TI - Suppressed Nna1 gene expression in the brain of ataxic Syrian hamsters. AB - Ataxic Syrian hamsters with an autosomal recessive trait were analyzed. Homozygotes showed moderate ataxia beginning at seven to eight weeks of age. They were fertile and lived more than two years. The affected hamsters exhibited an adult-onset degeneration of cerebellar Purkinje neurons, followed by a slow, mild reduction in the density of granule cells. Northern hybridization demonstrated that expression of Nna1, the gene responsible for the Purkinje cell degeneration (pcd) phenotype, was almost negligible in the brain of homozygous hamsters. These results strongly suggest that pcd-type mutation is involved in the ataxic phenotype of mutant hamsters. PMID- 17464796 TI - Screen for dominant behavioral mutations caused by genomic insertion of P-element transposons in Drosophila: an examination of the integration of viral vector sequences. AB - Here we report the development of a high-throughput screen to assess dominant mutation rates caused by P-element transposition within the Drosophila genome that is suitable for assessing the undesirable effects of integrating foreign regulatory sequences (viral cargo) into a host genome. Three different behavioral paradigms were used: sensitivity to mechanical stress, response to heat stress, and ability to fly. The results, from our screen of 35,000 flies, indicate that mutations caused by the random insertion of transposons in Drosophila are more effective at disrupting flight than stress sensitivity. This approach was used to ascertain the frequency of deleterious dominant mutations caused by viral vectors utilized in gene therapy. PMID- 17464797 TI - Effects of mutant Drosophila K+ channel subunits on habituation of the olfactory jump response. AB - The olfactory-jump response assay was used to analyze habituation in Drosophila mutants of potassium (K(+)) channel subunits. As with physiological assays of the giant fiber-mediated escape reflex, mutations at loci that encode K(+) channel subunits have distinct effects on habituating the olfactory-jump response. The data for slowpoke and ether a go-go indicate similar effects on habituation of the olfactory-jump response and the giant fiber-mediated escape. Habituation in the olfactory jump assay in Hyperkinetic and Shaker mutants was drastically different from the degree of defect in the giant fiber-mediated escape reflex, indicating differential control mechanisms underlying the two forms of non associative conditioning. PMID- 17464798 TI - The fickle mutation of a cytoplasmic tyrosine kinase effects sensitization but not dishabituation in Drosophila melanogaster. AB - fickle is a P-element mutation identified from a screen for defects in courtship behavior and disrupts the fly homolog of Bruton's tyrosine kinase (Btk) gene (Baba et al., 1999). Here, we show that habituation of the olfactory jump reflex also is defective in fickle. Unlike, the prototypical memory mutants, rutabaga and dunce, which habituate more slowly than normal, fickle flies habituate faster than normal. fickle's faster-than-normal response decrement did not appear to be due to sensorimotor fatigue, and dishabituation of the jump response was normal. Based on a long-standing "two opponent process" theory of habituation, these data suggested that behavioral sensitization might be defective in fickle. To test this hypothesis, we designed a olfactory sensitization procedure, using the same stimuli to habituate (odor) and dishabituate (vortexing) flies. Mutant flies failed to show any sensitization with this procedure. Our study reveals a "genetic dissection" of sensitization and dishabituation and, for the first time, provides a biological confirmation of the two opponent process theory of habituation. PMID- 17464800 TI - Training junior doctors. PMID- 17464801 TI - Acute fatty liver of pregnancy (AFLP)--an overview. PMID- 17464802 TI - Bartholin's cysts and abscesses. PMID- 17464803 TI - Surrogacy. PMID- 17464804 TI - Urinary tract infections in pregnant women with bacterial vaginosis. AB - Urinary tract infections (UTIs) and bacterial vaginosis are common problems in pregnancy and are associated with serious obstetric complications. We evaluated the risk of UTIs in pregnant women with bacterial vaginosis. A total of 322 pregnant women who presented for a prenatal visit were evaluated for bacterial vaginosis and UTIs, between March 2003 and September 2004. Bacterial vaginosis was diagnosed based on Amsel's criteria. Women using antibiotic or other vaginal cream, with specific vaginitis or vaginal bleeding were excluded. Data were analysed using chi(2), Student t-test and multiple logistic regression. A total of 76 women had bacterial vaginosis and 246 women did not; and 18 women (23.6%) with bacterial vaginosis had UTIs, compared with 24 (9.8%) of those without. Bacterial vaginosis was associated with an increased risk of UTIs (odds ratio (OR) 3.05; 95% CI: 1.47-6.33). Pregnant women with bacterial vaginosis are at increased risk for UTIs. PMID- 17464805 TI - A systematic review of experimental and clinical studies of sildenafil citrate for intrauterine growth restriction and pre-term labour. AB - Sildenafil could be an alternative in the treatment of intrauterine growth retardation (IUGR) and premature delivery. In order to systematically review the reproductive-related effects of sildenafil, a search was made on PubMed and the Science Citation Index for studies evaluating the effects of sildenafil on uterine vessels or myometrium either in vitro or in experimental animal models as well as for any clinical trial or case reporting the outcome of pregnant women treated with sildenafil. The information was obtained from: three in vitro studies, five studies performed in experimental animal models, four studies on women with fertility and sterility disorders receiving 100 mg/day of sildenafil intravaginally, and two case reports of pregnant women who received sildenafil for the treatment of pulmonary hypertension. Incubation with sildenafil of different in vitro preparations resulted in vasodilator and uterine relaxant effects. No evidence of teratogenicity was observed in the studies performed in mice, rats and dogs. Sildenafil increased fetal weight in rats. In women, contradictory results on uterine blood flow and endometrial development were reported after the intravaginal administration of sildenafil. No adverse fetal outcomes were reported in the two pregnant women with pulmonary hypertension receiving sildenafil late in their pregnancy. In conclusion, there is still limited information about the efficacy of sildenafil for the treatment of IUGR and premature delivery. However, studies in experimental animal models and two human case reports have reported no deleterious effects on the mother or offspring. PMID- 17464806 TI - Nifedipine serum levels in pregnant women undergoing tocolysis with nifedipine. AB - The objective of our cross-sectional, observational study was to investigate nifedipine serum levels in pregnant women undergoing tocolysis. A total of 24 pregnant women, 22-34 weeks' gestation, who were administered nifedipine for treatment of pre-term labour, were enrolled in the study. Blood samples were taken 12 h after the oral application of 60 mg nifedipine in 'continuous release' form (Adalat CR 60). Nifedipine serum levels were measured with liquid chromatography. Nifedipine serum levels spread between 6 and 101 ng/ml (17-292 nmol/l). There was no correlation between nifedipine levels and body mass index (BMI), or between nifedipine levels and gestational age. During nifedipine tocolysis, 11 of 24 patients (45.8%) had mild side-effects, mostly headache. The side-effects were not dose-related. Despite the standardised dosage and standardised blood sampling nifedipine serum levels spread in a wide range. There is no need to adjust the dose of nifedipine to BMI or to gestational age. PMID- 17464807 TI - Value of partogram in vaginal birth after caesarean section. AB - A prospective partographic study of labour in 102 women with one previous lower uterine segment caesarean section in the Aminu Kano Teaching Hospital, Kano, Nigeria, was carried out to assess the predictive value of partogram on the outcome of labour in vaginal birth after caesarean section (VBACS). The outcomes measured were the duration of labour, cervical dilatation rate (CDR), and average cervical dilatation rate (ACDR). The data obtained were analysed using Z-test and tests of validity. A successful vaginal delivery rate of 72.5% was achieved. The cervical dilatation rate, average cervical dilatation rate and the alert line were found to be significant predictors of the outcome of labour in VBACS. These findings have important implications in establishing policies for monitoring labour in VBACS. PMID- 17464808 TI - The peri-partum management of pregnancies with macrosomic babies weighing > or =4,500 g at a tertiary University Hospital. AB - The management of fetal macrosomia diagnosed antenatally presents a dilemma to the obstetrician. We retrospectively reviewed the peripartum management of singleton pregnancies, which ended in the delivery of a macrosomic baby (birth weight >/=4,500 g) in our unit between 1995 and 1999. This was to determine first, the associated maternal and neonatal morbidity and second, whether the lack of consensual management in our unit influences outcome. Over the 5-year period, there were 380 macrosomic births out of 26,974 deliveries; an incidence of macrosomia of 1.4%. The mean birth weight was 4,697 +/- 330 g (range 4,500 - 5,560 g). The onset of labour was spontaneous in 234 (61.6%) cases, 120 (31.6%) were inductions and 26 (6.8%) were elective caesarean sections. Of the 354 planned vaginal deliveries, 233 (65.8%) were spontaneous, 62 (17.5%) were operative vaginal deliveries and 59 (16.7%) were emergency caesarean sections. There was no relationship between the rate of successful vaginal delivery and birth weight. There were 40 (13.6%) cases of shoulder dystocia compared with 0.9% in the non-macrosomic population (p < 0.001). Emergency caesarean sections and shoulder dystocia were significantly more common with babies weighing >/=5,000 g (28.9% vs 15.2%, p < 0.002 for caesarean section and 25.8% vs 11%, p < 0.001 for dystocia). We therefore recommend that where the estimated fetal weight is >5,000 g, an elective caesarean section should be considered. Variations in the care provided by different consultants did not have any effect on outcome. Induction for fetal macrosomia alone did not improve outcome but was associated with a significantly higher emergency caesarean section rate and should therefore be discouraged. PMID- 17464809 TI - Pregnancy outcome in HIV-positive women in Enugu, Nigeria. AB - This is a retrospective case-control study of 62 HIV-positive women and 100 HIV negative controls who delivered in the University of Nigeria Teaching Hospital, Enugu, from 2 August, 2002 to 31 March, 2004. The HIV-positive women were relatively younger and of lower mean parity than the HIV-negative controls. They were also significantly more likely to have positive syphilis serology, higher mean duration of labour, perineal tear, puerperal sepsis and higher mean duration of hospital stay, higher prevalence of low birth weight, birth asphyxia and more admissions to the Newborn Special Care Unit than the controls (p < 0.05). However, there was no significant difference in the two groups in the prevalence of hepatitis B surface antigenaemia, recurrent vulvovaginitis, abortions, stillbirths, congenital anomalies, pre-term delivery, mean interval between rupture of membranes and delivery and mode of delivery (p > 0.05). All (100%) the HIV-negative and 96.8% of the seropositive women had voluntary counselling and testing (VCT). There was no maternal death in either group. Untreated maternal HIV infection is associated with adverse pregnancy outcomes in the form of increased maternal and fetal morbidities. Hence for optimal outcomes, prevention of mother-to-child transmission (PMTCT) programmes must incorporate combination drug treatment for the mother as early in pregnancy as possible. PMID- 17464810 TI - Maternal mortality in a rural referral hospital in the Niger Delta, Nigeria. AB - Almost two decades after the safe motherhood initiative, maternal mortality figures remain very high in Nigeria. Very few studies are available on the features of maternal mortality in rural Nigeria. The objective of this study was to determine the incidence and causes of maternal mortality in a rural referral hospital in the Niger Delta, Nigeria. An audit of 115 consecutive maternal mortalities over a 10-year period at a rural-based tertiary hospital was undertaken. There were 5,153 deliveries and 115 maternal deaths during the study period, with a maternal mortality ratio of 2,232/100,000 live births. The most common causes of maternal mortalities were puerperal sepsis, abortion complications, pre-eclampsia/eclampsia, prolonged obstructed labour, haemorrhage accounting for 33%, 22.6%, 17.4%, 13.0% and 7.8%, respectively. The percentage mortality for unbooked was 10 times that for booked patients. Unbooked status is a risk factor for maternal mortality as this was statistically significant p < 0.0001. Traditional birth attendants were involved in the initial management of at least two-fifths (38.2%) of the non-abortion mortalities while half had been managed in private hospitals and maternities. Maternal mortality will continue to increase unless appropriate steps are taken to improve the use of antenatal care, thereby reducing unbooked emergencies. Hospitals need to be equipped with facilities for emergency obstetric care. Continuous programmes that will integrate TBAs and orthodox practices should be put in place as this will reduce delays and improve referral systems. PMID- 17464811 TI - Adoption and practice of evidence-based obstetric care among Nigerian obstetricians. PMID- 17464812 TI - Long-term follow-up of patients with lichen sclerosus: does it really happen? AB - Lichen sclerosus is a common condition affecting the vulva and long-term follow up is recommended. We audited a group of patients discharged from the vulval clinic and found that 38% had not seen their GP 12 months after discharge. Of those that were seen, 17% were not examined. Further education is needed to improve the follow-up of these patients. PMID- 17464813 TI - Outpatient medical management of missed miscarriage using misoprostol. AB - We wanted to evaluate the efficacy, safety and acceptability of misoprostol for outpatient management of missed miscarriage. This was a prospective observational study conducted in the early pregnancy unit of a university teaching hospital involving women attending the unit with a diagnosis of missed miscarriage. All women with a missed miscarriage of <12 weeks' gestation and opting to have medical management were eligible for inclusion. Women who consented to take part were given 800 microg of misoprostol in the unit and advised to take two further doses of 400 microg of misoprostol, orally, 3 h apart, the following day at home. Outcome measures included success rates, emergency admission rates, blood transfusion rates, pain scores, satisfaction scores, incidence rates of side effects and number of women who would choose medical management in future. Of the 268 women diagnosed as having a missed miscarriage during the study period, 75 (28.0%) opted to have medical management. Over three-quarters (n = 58, 77.3%, 95% confidence interval (CI) 66.2 - 86.2) of the women achieved successful complete medical evacuation. Overall, five (6.7%; 95% CI 2.2 - 14.8) women presented for emergency admission. The mean visual analogue scale (VAS) score of all women was 6.7 (SD = 2.8) and the mean satisfaction score was 8.6 (SD = 2.1). Two-thirds of women (n = 50, 66.7%) said that they would choose to have medical evacuation in a future miscarriage, while seven (9.3%) were unsure. A total of 70 (93.3%) women said they preferred to have the treatment at home rather than in the hospital. We conclude that medical evacuation of missed miscarriage is efficacious, safe and acceptable in the outpatient setting. PMID- 17464814 TI - Uterine innervation in adenomyosis. AB - This study describes the innervation of the uterus with a histopathological diagnosis of adenomyosis in a retrospective survey of two groups of uteri. Group 1 consisted of 17 histologically-normal, parous uteri and eight nulliparous uteri. Group 2 consisted of 23 parous uteri with the histopathological diagnosis of adenomyosis. Tissue sections from the uterine isthmus were stained for nerves with PGP 9.5 using a standard immunohistochemical regimen. In group 1 (n = 25, normal histological report), normal innervation of the uterine isthmus included concentrations of nerves in the subserosal layers and at the endometrial myometrial interface with sparse, neurovascular bundles distributed throughout the myometrial stroma. In group 2, (n = 23 with adenomyosis), there were no nerves in areas of adenomyosis and absence of nerves at the endometrial myometrial nerve plexus. Focal proliferation of small-diameter nerve fibres was observed at the margins of adenomyosis in some uteri. Subserosal nerve fibres were still present in those sections that extended to include this region. Adenomyosis is associated with loss of nerve fibres at the endometrial-myometrial interface and absence of nerve fibres in the adenomyosis. PMID- 17464815 TI - Recurrent prolapse after sacrocolpopexy for post-hysterectomy vaginal vault prolapse. AB - This study looked at the subjective and objective long-term outcome of abdominal sacrocolpopexy for post-hysterectomy vaginal vault prolapse, with emphasis on recurrent prolapse. It included 42 patients who had abdominal sacrocolpopexy for post-hysterectomy vaginal vault prolapse from 1994 to 2004. Patients were invited for a follow-up visit to assess the objective and subjective outcome. The notes of 35 patients (83.3%) were available and 22 attended for follow-up (52.4%). None of them had a recurrent vault prolapse (0% direct failure rate). However, 20 cases (20%) had a repair operation by the time they were invited for follow-up and 18 of those attending for follow-up had anterior and/or posterior vaginal wall prolapse (81.8% indirect failure rate). Nonetheless, only eight of these (44.4%) were symptomatic, bringing the rate of indirect recurrences requiring management to 50%. Concomitant or prior repair operation was associated with a lower incidence of indirect recurrence. The incidence of urodynamic stress incontinence and detrusor overactivity was 13.6% and bowel emptying dysfunction was 4.5%. PMID- 17464816 TI - The value of cystoscopy and bladder biopsy taken at the time of tension-free vaginal tape insertion. AB - The primary aim of this study was to determine if a cystoscopy performed at the time of a tension-free vaginal tape (TVT) insertion is useful in finding unexpected pathology. A secondary outcome measure was to assess whether a bladder biopsy and cystoscopy performed during the TVT operation provide useful information about the outcome of surgery. The findings in 100 consecutive women were reviewed. A total of 42 women demonstrated mixed detrusor overactivity (DO) and urodynamic stress incontinence (USI); 32 women had abnormal biopsies. Women with DO were no more likely to have abnormal bladder biopsies than women without DO (p > 0.05). An abnormal bladder biopsy or cystoscopy failed to predict postoperative irritable symptoms (p > 0.05). The presence of preoperative DO correctly predicted postoperative irritable symptoms (p < 0.01). Cystoscopic abnormalities were noted in 14 women including one carcinoma, and one case of dysplasia was discovered on biopsy. In five women, the cystoscopy showed a bladder perforation. Cystoscopy should be performed in all women undergoing continence procedures. PMID- 17464817 TI - The menopause, hormone replacement therapy and informed consent: are women in an underresourced country adequately aware? AB - Recent evidence on the long-term effects of HRT have resulted in increased emphasis being placed on individualised counselling, patient choice and informed consent when managing the menopause. We assessed whether women in an underresourced country have adequate knowledge of the menopause/HRT to engage in patient - provider discussions and provide full informed consent for HRT. Specific 'knowledge scores' for the menopause and HRT were developed and utilised in structured questionnaires to determine the existing levels of knowledge in 150 women from different racial, educational and occupational backgrounds. Some 92% were aware of the menopause and 54% were aware of HRT. Specific knowledge about the menopause and HRT overall was low (39% and 38%, respectively). There was a significant association between higher education levels, race and occupational status on the knowledge of the menopause but not of HRT. Television, radio and pamphlets were the preferred sources to gain further information. There is a need to create awareness and provide further education to women in underresourced countries about the menopause and HRT to empower them to make informed choices about their health during this period. PMID- 17464818 TI - Participation in highly subsidized cervical cancer screening by women in Enugu, South-east Nigeria. AB - The current study was designed to evaluate the level of participation in a highly subsidized cervical screening in a resource-poor country. A total of 989 cervical smears performed on 932 women in Enugu, South-east Nigeria, over a 10-year period (January 1995-December 2004) was reviewed. The level of participation in cervical screening was very low, as <1% of the targeted women population participated. Almost 68% of the participants were referred for the screening and the majority (52.3%) were from lower social classes because the programme was highly subsidized. A total of 646 (65.3%) smears were normal. Of the abnormal smears, 193 (19.5%) had non-specific inflammatory changes, 136 (13.8%) showed dyskaryotic cells while 14 (1.4%) had neoplastic changes. A total of 57 (6.1%) women had more than one cervical cancer screening and they were characterised by increasing age, up to 59 years, higher social classes and contraceptive users in lower social classes. To reverse the low level of participation in cervical cancer screening in developing countries, there is a need to provide highly subsidized (if not free) cervical cancer screening services, which must be followed by sustained cervical cancer awareness campaign. PMID- 17464819 TI - Euglycaemic diabetic ketoacidosis in pregnancy: an unusual presentation. PMID- 17464820 TI - All that wheezes is not asthma. PMID- 17464821 TI - Chorea gravidarum. PMID- 17464822 TI - Cancer of the descending colon during pregnancy. PMID- 17464823 TI - Gastric volvulus: a rare cause of abdominal pain in pregnancy. PMID- 17464824 TI - Congenital brain tumour mimicking fetal intracranial haemorrhage. PMID- 17464825 TI - Familial exencephaly--anencephaly sequence and translocation. PMID- 17464826 TI - Numerous cord entanglements in a case of monoamniotic twin pregnancy. PMID- 17464827 TI - Shock associated ischaemic colitis following massive obstetric haemorrhage. PMID- 17464828 TI - Pseudocyesis in a teenager using long-term contraception. PMID- 17464829 TI - Hyperventilation with medroxyprogesterone therapy in a postmenopausal woman. PMID- 17464830 TI - Thermal balloon endometrial ablation in bicornuate uteri. PMID- 17464831 TI - Post-hysterectomy fallopian tube prolapse--a diagnostic pitfall. PMID- 17464832 TI - Vagino-rectal fistula caused by Bartholin's abscess. PMID- 17464833 TI - A rare cause for cervical pigmentation. PMID- 17464834 TI - Spontaneous ovarian abscess associated with an endometrioma. PMID- 17464835 TI - Cystic cavernous lymphangioma of the mesentery in a patient with Cowden syndrome. PMID- 17464836 TI - Pelvic kidney mistaken for multicystic ovary at ultrasound scan. PMID- 17464837 TI - Primary vaginal adenocarcinoma of intestinal type arising from an adenoma. PMID- 17464838 TI - Lipoleiomyosarcoma of the vagina. PMID- 17464840 TI - ITS barcodes for Trichophyton tonsurans and T. equinum. AB - Early molecular biosystematic studies of dermatophytes created considerable confusion about the taxonomic status of the horse-associated Trichophyton equinum vis-a-vis the anthropophilic T. tonsurans. Though this matter has recently been clarified, routine identification of these species based on the commonly used ribosomal internal transcribed spacer (ITS) sequence has been impractical. This is because, in the available sequences attributed to the species in GenBank, a clear species-level distinction does not appear to exist. In the present study, resequencing the ITS regions of several anomalous isolates is shown to eliminate this problem, which was mainly based on read errors in older sequences. Newly generated sequences and recent GenBank additions are analysed to show that T. equinum appears to be uniform in ITS sequence worldwide, while T. tonsurans is also uniform, excepting a single-base change found in one otherwise typical strain. Analysis also reveals a distinct, as yet incompletely classified Asian genotype that may belong to one or the other of these species. Standard ITS 'barcode sequences' are proposed for T. tonsurans and T. equinum, and a taxonomic neotype is designated to anchor the latter species. T. equinum var. autotrophicum is further evidenced as very closely related to T. equinum var. equinum, and the anomaly of its plesiomorphous phenotype is discussed in a population genetics context. PMID- 17464841 TI - Occurrence of Scopulariopsis and Scedosporium in nails and keratinous skin. A 5 year retrospective multi-center study. AB - A 5-year retrospective multicenter study was performed for microascaceous moulds (Microascaceae, Ascomycetes) in Finnish clinical specimens. The files from 1993 1997 of six clinical mycology laboratories in Finland were searched for reports of these fungi, mainly Scopulariopsis and Scedosporium anamorphs in keratinous specimens. From the 521 primary findings, 165 cases were selected for further study based on direct microscopy, colony numbers and accompanying fungi. The clinical records of 148 cases (141 Scopulariopsis, 7 Scedosporium) were studied. Of the nail infections from which Scopulariopsis was recovered, 39 cases were further separated which showed clinical or laboratory-based evidence of dermatophytosis. In the remaining 90 'non-dermatophyte' nail cases, Scopulariopsis spp. were the only documented fungal agents (c. 6 cases/million/year). The patients were mainly elderly, 66% of whom had problems involving their big toe nails. For 74% of them, the nail problem was mentioned as their reason for visiting the physician. However, only 18% had documented benefit from treatment. The Scopulariopsis nail infections seem to be treatment-resistant and the pathogenesis and etiological role of Scopulariopsis remain poorly understood. PMID- 17464842 TI - Targeting polyamines of Aspergillus nidulans by siRNA specific to fungal ornithine decarboxylase gene. AB - Aspergillus nidulans is a filamentous, ubiquitous and opportunistic pathogenic fungus, which causes fatal invasive aspergillosis among immunocompromised patients. As a prelude to the investigations on the possible control of human fungal diseases by selective targeting of fungal polyamines that are essential for fungal growth and development by RNAi strategy, we have examined the effect of siRNA specific to a key polyamine biosynthesis gene ornithine decarboxylase (ODC) on the in vitro germinating spores of A. nidulans to assess the ability of RNA species to induce RNA-mediated gene silencing. The present findings show that siRNA can cause specific silencing effect, with the phenotypic consequences leading to significant reduction in mycelial growth, target mRNA titers and cellular polyamine concentrations. This study suggests that fungal polyamine biosynthetic genes, particularly ODC may be targeted for the control of fungal infections in humans and crop plants. To the best of our knowledge, this is the first demonstration of up-take of siRNA by germinating fungal spores from culture medium, and attempt to target a vital pathway in fungal pathogens for their control. PMID- 17464843 TI - Therapeutic efficacy of posaconazole against isolates of Candida albicans with different susceptibilities to fluconazole in a vaginal model. AB - A battery of 34 vaginal isolates of Candida albicans was tested against posaconazole (POS) and fluconazole (FLU) to determine their in vitro susceptibilities and to obtain FLU-susceptible and FLU-resistant strains for the murine in vivo studies. FLU-resistant strains were chosen on the basis of their 48-h MICs. The 48-h geometric mean MICs for all isolates tested were 0.016 and 0.656 microg/ml for POS and FLU, respectively. The treatment regimens for the vaginal murine infection model were POS or FLU at 10 or 20 mg/kg of body weight/day and 20 mg/kg twice a day. All regimens with POS were effective in reducing fungal burden of both the fluconazole-susceptible and resistant isolates of C. albicans. All FLU regimens were effective against infection induced by the fluconazole-susceptible strain. While FLU at 10 mg/kg was ineffective against fungal burden of the resistant strain, treatment with FLU at 20 mg/kg once or twice a day was effective against this strain. Both POS and FLU at 20 mg/kg twice a day were able to clear C. albicans from vaginas of mice infected with the fluconazole-susceptible strain. POS displayed a more effective in vivo activity than FLU in the treatment of murine C. albicans vaginitis produced by isolates with different susceptibilities to FLU. PMID- 17464844 TI - Production of mycotoxins by Aspergillus lentulus and other medically important and closely related species in section Fumigati. AB - The production of mycotoxins and other secondary metabolites have been studied by LC-DAD-MS from six species in Aspergillus section Fumigati. This includes the three new species Aspergillus lentulus, A. novofumigatus and A. fumigatiaffinis as well as A. fumigatus, Neosartoria fisheri and N. pseudofisheri. A major finding was detection of gliotoxin from N. pseudofisheri, a species not previously reported to produce this mycotoxin. Gliotoxin was also detected from A. fumigatus together with fumagillin, fumigaclavine C, fumitremorgin C, fumiquinazolines, trypacidin, methyl-sulochrin, TR-2, verruculogen, helvolic acid and pyripyropenes. Major compounds from A. lentulus were cyclopiazonic acid, terrein, neosartorin, auranthine and pyripyropenes A, E and O. Thus in the present study A. fumigatus and A. lentulus did not produce any of the same metabolites except for pyripyropenes. The fact that A. lentulus apparently does not produce gliotoxin supports the idea that other compounds than gliotoxin might play an important role in the effective invasiveness of A. lentulus. An overall comparison of secondary metabolite production by strains of the six species was achieved by analysis of fungal extracts by direct injection mass spectrometry and cluster analysis. Separate groupings were seen for all the six species even though only one isolate was included in this study for the two species A. novofumigatus and A. fumigatiaffinis. PMID- 17464845 TI - Molecular epidemiology of canine histoplasmosis in Japan. AB - A recent case of canine histoplasmosis, the first confirmed case of disseminated infection accompanied by carcinoma in Japan, was diagnosed by clinical characteristics, histopathological examination, chest radiographs, ocular fundoscopy and molecular biological data. The clinical manifestations were not limited to cutaneous symptoms but were referable to disseminated infection, similar to human autochthonous cases. The partial sequences of the internal transcribed spacer (ITS1/2) regions of the ribosomal DNA genes of this and other Japanese canine histoplasmosis strains were 99-100% identical to the sequence AB211551 derived from a human isolate in Thailand, and showed a close relationship to the sequences derived from Japanese autochthonous systemic and cutaneous human cases. The phylogenetic analysis of 97 sequences of the ITS1/2 region disclosed six genotypes. The genotypes derived from Japanese autochthonous human and dog cases belonged to the cluster consisting of Histoplasma capsulatum var. capsulatum and H. capsulatum var. farciminosum sequences, indicating that these varieties might cause not only cutaneous but also systemic histoplasmosis, regardless of their host species. The current status of the 3 varieties of Histoplasma capsulatum according to the host species remains a subject of further investigation. PMID- 17464846 TI - Protothecosis in 17 Australian dogs and a review of the canine literature. AB - Systemic protothecosis was diagnosed in 17 Australian dogs between 1988 and 2005. There was a preponderance of young-adult (median 4 years), medium- to large-breed dogs. Females (12/17 cases) and Boxer dogs (7 cases, including 6 purebreds and one Boxer cross) were over-represented. Sixteen of 17 dogs died, with a median survival of four months. A disproportionate number of cases were from coastal Queensland. In most patients, first signs were referable to colitis (11/17 cases), which varied in severity, and was often present for many months before other symptoms developed. Subsequent to dissemination, signs were mostly ocular (12 cases) and/or neurologic (8 cases). Two dogs had signs due to bony lesions. Once dissemination was evident, death or euthanasia transpired quickly. Prototheca organisms had a tropism for the eye, central nervous system (CNS), bone, kidneys and myocardium, tissues with a good blood supply. Microscopic examination and culture of urine (5 cases), cerebrospinal fluid (CSF;1 case), rectal scrapings (4 cases), aspirates or biopsies of eyes (5 cases) and histology of colonic biopsies (6 cases) as well as skin and lymph nodes (2 cases) helped secure a diagnosis. Of the cases where culture was successful, P wickerhamii was isolated from two patients, while P zopfii was isolated from five. P zopfii infections had a more aggressive course. Treatment was not attempted in most cases. Combination therapy with amphotericin B and itraconazole proved effective in two cases, although in one of these treatment should have been for a longer duration. One surviving dog is currently still receiving itraconazole. Protothecosis should be considered in all dogs with refractory colitis, especially in female Boxers. PMID- 17464847 TI - Lecythophora hoffmannii isolated from a case of canine osteomyelitis in Japan. AB - A 2-year-old spayed female mongrel dog showed claudication with abnormal ossification containing fungal cells detected by biopsy. The dog was treated with ketoconazole and itraconazole perorally for 5 months; however, the osteomyelitis became aggravated, and an amputation from the scapula was performed. The right superficial cervical lymph node became swollen 5 months after the operation. The lymph node contained PAS positive fungal elements and a portion of tissue produced mycelial fungal growth on potato dextrose agar supplemented with chloramphenicol. The culture was identified as Lecythophora hoffmannii based on morphology, physiology and 100% identity in the sequence of the D1/D2 domain of the large subunit ribosomal RNA gene of the fungal species in the GenBank database (accession number AB100627). In addition, the sequence from the present isolate was submitted as AB189164. The isolate showed resistance to antifungal agents, i.e., amphotericin B, 5-FC, fluconazole, itraconazole, miconazole and micafungin. The dog developed cachexia 2 months after the onset of lymphadenopathy, and was euthanized on the 459th day after onset of clinical symptoms. This was the first disseminated case of L. hoffmannii infection in Japan. PMID- 17464848 TI - Clinical efficacy of micafungin for chronic pulmonary aspergillosis. AB - The rising incidence of pulmonary aspergillosis is a major clinical concern. However, only a limited number of antifungal drugs are available in Japan that are effective for pulmonary Aspergillus infections. Micafungin (MCFG), a newly developed echinocandin family antifungal drug, has potent antifungal activity in vitro, but few reports detailing its clinical effectiveness have been published to date. A retrospective study was performed using data from nine patients (seven males and two females) with chronic invasive forms of pulmonary aspergillosis, who were treated with either MCFG alone or together with other antifungal drugs between April 2003 and March 2004. The overall efficacy of the treatments was evaluated in the terms of clinical, mycological, serological and radiological responses. The average age of the patients was 61.9 (20-83) years old. Four patients received only MCFG and the remaining five patients were treated with MCFG in combination with amphotericin B (AMB) only (1 patient), itraconazole (ITC) only (2 patients) or AMB backed up by ITC during AMB discontinuation periods (2 patients). The mean duration of MCFG administration was 59.2 (28-96) days. Overall, the treatment was judged to have been effective for seven of nine patients. No patient's condition deteriorated in response to treatment. Administration of MCFG alone was judged to have been effective in three of four patients. No notable adverse effects were documented during MCFG administration. These data suggest that MCFG may be an effective and safe antifungal drug for the treatment of chronic invasive forms of pulmonary aspergillosis. PMID- 17464850 TI - Newly described Aspergillus species within the section Fumigati. PMID- 17464849 TI - First Spanish case of onychomycosis caused by Chaetomium globosum. AB - Members of the fungal genus Chaetomium usually colonize cellulose-containing plant remains but on rare occasions may cause opportunistic mycoses and cutaneous infection in otherwise healthy individuals. To our knowledge, there have been only five credible descriptions of onychomycosis caused by members of this genus and only two of these contained information on therapy. We describe the first case of Chaetomium globosum onychomycosis recorded in Spain. The etiologic significance of the fungus was confirmed by its repeated isolation at different times, to the exclusion of dermatophytes. Clinically, the affected nails showed an excellent response to terbinafine and complete cure appeared to have been attained. PMID- 17464851 TI - Factors affecting the epidemiology of Acanthamoeba keratitis. AB - Despite being a relatively rare disease in comparison with other forms of infectious keratitis, Acanthamoeba keratitis is a potentially blinding disease. Wide variations in the incidence of Acanthamoeba keratitis have been reported in both developed and developing countries. At the same time that contact lens wear was found to be responsible for the spread of the disease in developed countries, Acanthamoeba keratitis was considered a rare disease in developing countries compared with fungal and bacterial keratitis. In recent decades, the risk of getting Acanthamoeba keratitis has increased because of the increased proportion of contact lens wearers. This article introduces the different factors affecting the epidemiology of Acanthamoeba keratitis worldwide, presents a chronological review of the literature, and shows the progressive spread of Acanthamoeba keratitis in the last two decades in different geographical areas of the world. A detailed comparison of the incidence of the disease as reported in different studies in different countries is made. The impact of contact lenses and other factors, such as hot weather, virulence of Acanthamoeba strains, water sanitation and quality, the occurrence of environmental disasters such as flooding, and the wide environmental presence of Acanthamoeba cysts on the incidence of the disease, are discussed. In addition, the ability of Acanthamoeba cysts to resist different harsh conditions is reviewed. PMID- 17464852 TI - Microbial keratitis in South India: influence of risk factors, climate, and geographical variation. AB - PURPOSE: To determine the influence of risk factors, climate, and geographical variation on the microbial keratitis in South India. METHODS: A retrospective analysis of all clinically diagnosed infective keratitis presenting between September 1999 and August 2002 was performed. A standardised form was filled out for each patient, documenting sociodemographic features and information pertaining to risk factors. Corneal scrapes were collected and subjected to culture and microscopy. RESULTS: A total of 3,183 consecutive patients with infective keratitis were evaluated, of which 1,043 (32.77%) were found to be of bacterial aetiology, 1,095 (34.4%) were fungal, 33 (1.04%) were Acanthamoeba, 76 (2.39%) were both fungal and bacterial, and the remaining 936 (29.41%) were found to be culture negative. The predominant bacterial and fungal pathogens isolated were Streptococcus pneumoniae (35.95%) and Fusarium spp. (41.92%), respectively. Most of the patients (66.84%) with fungal keratitis were between 21 and 50 years old, and 60.21% of the patients with bacterial keratitis were older than 50 (p < 0.0001) (95% CI: 5.19-7.19). A majority of patients (64.75%) with fungal keratitis were agricultural workers (p < 0.0001) [odds ratio (OR): 1.4; 95% CI: 1.19-1.61], whereas bacterial keratitis occurred more commonly (57.62%) in nonagricultural workers (p < 0.0001) (OR: 2.88; 95% CI: 2.47-3.36). Corneal injury was identified in 2,256 (70.88%) patients, and it accounted for 92.15% in fungal keratitis (p < 0.0001) (OR: 7.7; 95% CI: 6.12-9.85) and 100% in Acanthamoeba keratitis. Injuries due to vegetative matter (61.28%) were identified as a significant cause for fungal keratitis (p < 0.0001) (OR: 23.6; 95% CI: 19.07-29.3) and due to mud (84.85%) for Acanthamoeba keratitis (p < 0.0001) (OR: 26.01; 95% CI: 3.3-6.7). Coexisting ocular diseases predisposing to bacterial keratitis accounted for 68.17% (p < 0.0001) (OR: 33.99; 95% CI: 27.37 42.21). The incidence of fungal keratitis was higher between June and September, and bacterial keratitis was less during this period. CONCLUSION: The risk of agricultural predominance and vegetative corneal injury in fungal keratitis and associated ocular diseases in bacterial keratitis increase susceptibility to corneal infection. A hot, windy climate makes fungal keratitis more frequent in tropical zones, whereas bacterial keratitis is independent of seasonal variation and frequent in temperate zones. Microbial pathogens show geographical variation in their prevalence. Thus, the spectrum of microbial keratitis varies with geographical location influenced by the local climate and occupational risk factors. PMID- 17464853 TI - Histopathological diagnosis of corneal button specimens: an epidemiological study. AB - PURPOSE: To determine the most common histopathological diagnosis of corneal specimens from penetrating keratoplasty (PKP). METHODS: The records of 500 corneal specimens submitted to biopsy at the Henry Witelson Ocular Pathology Laboratory, Montreal, Canada, from 1999 to 2004 were reviewed. Age, sex, clinical indications, and histopathological findings were analyzed. RESULTS: Chronic keratitis (45.6%) was the most common pathological diagnosis, followed by corneal edema (25.8%), dystrophy (12.8%), keratoconus (KC) (9.2%), acute keratitis (5.6%), and degeneration (1.0%). Among the specimens with chronic keratitis, regraft was the most common clinical indication (39.0%). In the group of acute keratitis, ulcerative condition was the leading cause (75,0%). Fuchs' endothelial dystrophy represented 79.7% of the clinical diagnoses in the group of corneal dystrophies. The median patient age was 70-79 years, and the gender distribution was nearly symmetric. CONCLUSION: The present study is important for determining the most common histopathological diagnoses of corneal button specimens and the correlation with the age, gender, and clinical indications of PKP. PMID- 17464854 TI - Duration of diabetes and screening coverage for retinopathy among patients with type 2 diabetes. AB - AIMS: Despite reporting of the Wisconsin Epidemiologic Study of Diabetic Retinopathy(1) and the Diabetic Retinopathy Awareness Program(2) that diabetes duration was a significant predictor for adherence to vision care guidelines, reports of estimates of screening coverage for diabetic retinopathy taking into account diabetes duration have been lagging. This article estimates considering diabetes duration, the prevalence of diabetic retinopathy and screening coverage for diabetic retinopathy among type 2 diabetic patients. METHODS: As part of a treatment program at a High-Resolution Diabetes Center in Spain, type 2 diabetic patients attending the center from January 2003 to January 2005 were invited to participate in the study. Data on age, sex, and diabetes were recorded into a questionnaire, as was information about previous eye examinations. Polaroid(R) photographs were taken of the eye fundus with the poorest visual acuity using a nonmydriatic retinal camera. RESULTS: A total of 217 type 2 diabetic patients entered the program. The average age and duration of diabetes was 60.9 years and 7 years, respectively. Screening coverage for diabetic retinopathy was higher in those with a longer duration of diabetes (chi(2) = 36.5; p = 0.001). Fifty percent of patients had developed some retinopathy within the first 5 years after the diagnosis of the disease, but only 26.1% had received a previous fundus examination. CONCLUSIONS: These results argue for screening programs for people with type 2 diabetes mellitus focused on the subgroup of patients with diabetes duration of 5 years or less. PMID- 17464855 TI - Self-reported visual impairment and mortality: a French nationwide perspective. AB - PURPOSE: To estimate the association between self-reported visual impairment and mortality. METHODS: Two national surveys in community and institutionalized populations were combined. First, 2,075 institutions for children with impairments, adults with impairments aged persons, and psychiatric patients were selected randomly. The sample comprised 15,403 subjects of whom 14,603 (94.9%) were interviewed. Second, a random, stratified sample of 21,760 persons living in the community was selected, and 16,945 (77.9%) were interviewed. Types of impairment were identified by face-to-face interviews. Two years later, 14,497 subjects in institutions and 15,648 in the community were revisited. Data on death were obtained from either the National Register or households. Death rates were related to age, gender, and impairment. A logistic regression was performed including impairments, activities of daily living, age, gender, type of residence, and geographical area. RESULTS: Strong, independent associations were found between particular impairments, institutional residence, activities of daily living, age, gender, and risk of death. Associations between mortality and type of impairment could be ranked as follows: motor (OR = 1.235), brain (OR = 1.552), low vision (OR = 1.681), speech (OR = 2.090), visceral (OR = 2.233) and blindness (OR = 2.262). CONCLUSIONS: Self-reported visual impairment is an independent factor associated with mortality. PMID- 17464856 TI - Can information on the purpose of spectacle use and age at first use predict refractive error type? AB - PURPOSE: To assess the sensitivity and specificity of predicting refractive error type using information from a four-item questionnaire on the purpose of spectacle use and age at first use. METHODS: The Sydney Myopia Study examined 1,740 year 1 (78.9% response) and 2,353 year 7 students (75.3% response) from a random cluster sample of 34 primary and 21 secondary schools across Sydney. Parents of participants completed a four-item questionnaire that sought data on parental spectacle use, age at first use, and purpose of use (for clear distant vision, close work, or both). Prescriptions were obtained for 720 of 3,209 (22%) parents (73% of those approached) for validation. A receiver operating characteristic (ROC) curve was used to determine the optimal cutoff age for spectacle use in myopia classification. RESULTS: Using the ROC curve, a cutoff age of 30 years at first spectacle use produced the highest accuracy in determining myopia. We combined information on the purpose for using spectacles (for distant and near vision) and age of first use at 30 years or younger to determine myopia, otherwise hyperopia. Validated against prescriptions, the sensitivity and specificity of these predictions were 0.89 and 0.83, respectively, for myopia. The specificity was 0.92 for hyperopia and 0.80 for astigmatism, though corresponding sensitivities were lower at 0.23 and 0.46, respectively. CONCLUSIONS: In a sample of the parents of Sydney Myopia Study participants, information on the purpose of spectacle use with an age-at-first-use criterion can identify myopic refractive error with reasonable sensitivity and specificity. This four-item questionnaire may assist future epidemiological studies of screening for myopia. PMID- 17464857 TI - Current indications and resultant complications of evisceration. AB - PURPOSE: Evisceration is an alternative treatment modality to enucleation for many end-stage eye diseases. No study has addressed the indications for evisceration of eyes in Saudi Arabia. The aim of this study was to determine the current clinical indications for evisceration in patients at a tertiary eye care center and attempt clinicopathological correlation. METHODS: Clinical records of patients who had undergone evisceration at a tertiary eye care center over a 4 year period were reviewed retrospectively. The patients' demographic data and clinical indications for evisceration were studied, and the results from histopathological findings were correlated with the clinical diagnosis. RESULTS: Evisceration of the eyes was performed in 187 patients. Males outnumbered females in a ratio of 1.3:1 (105 males and 82 females). Blind painful eye was the primary presenting symptom in 117 (62.6%) patients and unsightly eye in 38 (20.3%) patients. Clinical indications for evisceration included endophthalmitis in 85 (45.5%), phthisis bulbi in 38 (20.3%), traumatic injury in 36 (19.2%), and glaucoma in 14 (7.5%) patients. Sixty-three patients (33.7%) had prior history of cataract surgery, penetrating keratoplasty, glaucoma surgery, or retina surgery. Clinicopathological correlation was 100% in cases with definite clinical diagnosis of endophthalmitis. CONCLUSION: Blind painful eye, endophthalmitis, phthisis bulbi, severe traumatic injury, and glaucoma were the major indications for eviscerations in a tertiary eye care center. PMID- 17464858 TI - Congenital color blindness in young Turkish men. PMID- 17464860 TI - Pregnancy complications and birth outcomes of pregnant women with urinary tract infections and related drug treatments. AB - Maternal urinary tract infections in pregnancy showed an association with a higher rate of preterm birth in previous studies. The aim of this study was to check this relationship, and in addition to evaluate the efficacy of recent medical treatments. The population-based large control (without any defects) data set of the Hungarian Case-Control Surveillance System of Congenital Abnormalities was evaluated. Of 38,151 newborn infants, 2188 (5.7%) had mothers with urinary tract infections during pregnancy, and 90% of these maternal diseases were prospectively and medically recorded. The prevalence of pre-eclampsia and polyhydramnios showed an association with urinary tract infections during pregnancy. Pregnant women with urinary tract infections in pregnancy had a somewhat shorter gestational age (0.1 week) and a higher proportion of preterm births (10.4% vs 9.1%). These differences were correlated with the severity of urinary tract infections. However, the preterm-inducing effect of maternal urinary tract infections is preventable by some antimicrobial drugs such as ampicillin, cefalexin and cotrimoxazole. In conclusion, maternal urinary tract infections during pregnancy increase pre-eclampsia and polyhydramnios, and in addition the rate of preterm birth; however, the latter is preventable by appropriate drug treatments. PMID- 17464861 TI - Listeriosis in Norway 1977-2003. AB - In Europe, the incidence of invasive listeriosis has increased substantially during the last decades. We here present data from 289 listeriosis cases reported in Norway during the period 1977-2003, of which 12 cases were associated with 2 outbreaks and 39 cases were pregnancy-related. Medical records were obtained from 209 cases with listeriosis reported in 1977-2000. While the incidence of pregnancy-related listeriosis has remained stable at an average rate of 34 per million pregnant women per y during the period, the incidence of sporadic, non pregnancy-related cases has increased from 1.1 to 3.7 per million per y. The present Norwegian incidence of reported cases is lower than in Denmark, but the case fatality rate is higher, indicating a possible under-reporting of mild listeriosis cases in Norway. We discuss how preventive measures, case identification and surveillance may have influenced listeriosis incidence in Norway. PMID- 17464862 TI - Evaluation of an automated extraction method for the detection of Chlamydia trachomatis and Neisseria gonorrhoeae by Cobas Amplicor PCR from different sample materials. AB - A commercially available automated device (MagNA Pure LC) was adapted for nucleic acid extraction of urogenital specimen for subsequent PCR detection of Chlamydia trachomatis and Neisseria gonorrhoeae in a clinical laboratory. Results were compared to the standard manual extraction procedure and showed excellent correlation, with even slightly increased sensitivity. PMID- 17464863 TI - Helicobacter pylori-associated chronic idiopathic thrombocytopenic purpura and low molecular weight H. pylori proteins. AB - We examined the association between Helicobacter pylori infection and chronic idiopathic thrombocytopenic purpura (cITP) to clarify the development of H. pylori-associated cITP. cITP patients were classified into 3 different groups: Hp negative (HP-N); Hp-positive, completely or partially responsive to treatment (CR); and Hp-positive and unresponsive to treatment (NR). Reactivity of antibodies to H. pylori before and after eradication was examined by immunoblotting. We used immunoblotting with immunoprecipitation to establish whether platelets form complexes with H. pylori proteins and if these complexes react with patients' sera. CR group showed large (>50 kDa) and low molecular weight protein bands, especially of 36, 27 and 17-kDa. These low molecular weight bands were detected significantly more in the CR group compared to other groups. When healthy human platelets were incubated with H. pylori lysate, they aggregated with the lysate, indicating that complexes were formed between the platelets and the lysate. The complexes immunoprecipitated with anti-human thrombocyte antibodies, and showed a 17 kDa band in the CR, but not in other groups. At least 3 low molecular weight proteins of H. pylori were involved in H. pylori-associated cITP. Immunocomplexes consisting of platelets, low molecular weight proteins of H. pylori and anti-H. pylori antibodies may represent an extra mechanism in development of H. pylori- associated cITP. PMID- 17464864 TI - Detection and molecular genetics of extended-spectrum beta-lactamases among cefuroxime-resistant Escherichia coli and Klebsiella spp. isolates from Finland, 2002-2004. AB - Extended-spectrum beta-lactamase (ESBL) producing Escherichia coli and Klebsiella spp. isolates are spreading and becoming an increasing problem concerning treatment, diagnostics and hospital hygiene. We wanted to discover which genotypes are occurring in Finland and to assess the CLSI screening method. The isolates were collected from 26 laboratories during a 3-y period from 2002 to 2004. We studied the zone diameters by disk diffusion according to CLSI recommendations. ESBL genes were detected by PCR and the TEM and SHV genes were sequenced traditionally, while the CTX-M isolates were analysed with pyrosequencing. Of the 402 isolates included in the study, 269 (67%) were confirmed to be ESBL producers according to the CLSI criteria. The CTX-M genes were the most prevalent, especially the combination of a CTX-M-1-group and a TEM 1 gene. In our material there were few isolates that had an ESBL gene but were negative in the CLSI ESBL confirmatory test. During recent y especially the CTX-M producing isolates have increased in Europe and now they are also found in Finland with increasing prevalence. PMID- 17464865 TI - Acute facial nerve palsy in children: how often is it lyme borreliosis? AB - Acute facial nerve palsy in children may be caused by infection by Borrelia burgdorferi, but the incidence of facial nerve palsy and the proportion of facial nerve palsy caused by Lyme borreliosis may vary considerably between areas. Furthermore, it is not well known how often facial nerve palsy caused by Lyme borreliosis is associated with meningitis. In this population-based study, children admitted for acute facial nerve palsy to Stavanger University Hospital during 9 y from 1996 to 2004 were investigated by a standard protocol including a lumbar puncture. A total of 115 children with facial nerve palsy were included, giving an annual incidence of 21 per 100,000 children. 75 (65%) of these were diagnosed as Lyme borreliosis, with all cases occurring from May to November. Lymphocytic meningitis was present in all but 1 of the children with facial nerve palsy caused by Lyme borreliosis where a lumbar puncture was performed (n = 73). In this endemic area for Borrelia burgdorferi, acute facial nerve palsy in children was common. The majority of cases were caused by Lyme borreliosis, and nearly all of these were associated with lymphocytic meningitis. PMID- 17464866 TI - Tigecycline: its potential for treatment of brucellosis. AB - The in vitro efficacy and synergistic activity of tigecycline in comparison with other antimicrobials used in brucellosis, were tested for 16 Brucella melitensis strains by the E-test method. Tigecycline had the lowest minimal inhibitory concentration levels, and rifampin the highest, in the study. Tigecycline also provided the better synergistic activity compared to doxycycline according to the fractional inhibitory concentration index. The results of this in vitro study suggest tigecycline as a therapeutic alternative for brucellosis. These observations need to be supported with clinical studies. PMID- 17464867 TI - Application of an ELISA test using Schistosoma bovis adult worm antigens in travellers and immigrants from a schistosomiasis endemic area and its correlation with clinical findings. AB - We have recently evaluated an ELISA for the diagnosis of human schistosomiasis using S. bovis adult worm antigens (AWA Sb), showing a sensitivity of 94% and a specificity of 97% for patients diagnosed by egg detection. Nevertheless, the comparison of this AWA Sb ELISA with direct parasitological findings as the gold standard could introduce a selection bias, due to the well-known lack of sensitivity of direct methods in the detection of acute schistosomiasis and of low burden infections. The objective of the present work is to compare it with parasitological methods and commercial indirect haemagglutination test using S. mansoni antigens (WA Sm IHA) in 254 immigrants and travellers with different clinical settings; in addition, to find specific bands in the EITB of different phases of schistosomiasis. The AWA Sb ELISA showed 72% of seropositivity in patients with Katayama fever, while patients with eosinophilia and genito-urinary complaints showed 27% and 93%, respectively. The diagnosis yield was globally higher than direct egg detection or WA Sm IHA test with regard to the clinical setting. Finally, the utilization of EITB with S. bovis AWA permits the confirmation of diagnosis in chronic and acute phases of the disease. PMID- 17464868 TI - Analyses of Bcl-2, Survivin, and CD44v6 expressions and human papillomavirus infection in cervical carcinomas. AB - Infection with human papillomaviruses (HPV), and suppression of apoptosis and cell adhesion are putative aetiological factors to cervical carcinogenesis. However, controversial results have been reported with respect to their relationships with cervical carcinomas. Here we analysed papillomavirus infection, apoptotic index (AI), expressions of the anti-apoptotic proteins Bcl-2 and Survivin, and expression of the cell-adhesion protein CD44 in cervical tissue samples from individuals with and without cervical carcinomas. Although both HPV16 and HPV18 are reportedly important aetiological factors, we found that cervical carcinomas were highly associated with HPV16 but not HPV18 infection. Immunohistochemistry showed that the percentages of cells expressing Bcl-2, Survivin, and CD44v6 were greatly increased in samples of cervical carcinomas. Furthermore, the expression rates of Survivin and CD44v6 increased whereas that of Bcl-2 declined as cervical cancers developed into more advanced clinical or histopathological stages. Surprisingly, there was little difference in AI between control and cervical cancer samples. These observations provide further evidence that HPV infection, apoptosis and cell adhesion abnormalities are related to cervical cancers. They also suggest that Bcl-2, Survivin and CD44v6 expressions, and HPV16 infection could be useful indices in screening of cervical carcinomas. PMID- 17464869 TI - Fatal intrauterine infection by herpes simplex virus type 2 in an infant from a mother lacking seroreactivity to glycoprotein G. AB - In this case report we describe a fatal intrauterine HSV-2 infection in a mother with insulin-dependent diabetes mellitus without signs, symptoms or serological evidence of infection with this virus during pregnancy. A normally developed infant was delivered by caesarean section and the course of the viral infection was rapid and fatal within the first d. Histopathology demonstrated disseminated intravascular coagulation in several organs. The diagnosis was confirmed by PCR amplifications of HSV-2 DNA from several organs of the child at autopsy and further supported by DNA sequencing of the viral amplicon derived from brain. Despite a significant IgG titre rise to a type-common HSV IgG antigen being seen in the mother, no IgG response to the HSV-2 type-specific glycoprotein G (gG) could be documented during long-term follow-up. PMID- 17464870 TI - Locally acquired hepatitis E in the Netherlands: associated with the consumption of raw pig meat? AB - We here present 2 patients who developed hepatitis E, without having been abroad or in contact with anyone who did, indicating locally acquired hepatitis E. We point out that the consumption of raw pig meat could be of relevance in HEV associated hepatitides in the Netherlands. PMID- 17464871 TI - Persistent bacteraemia due to methicillin-resistant Staphylococcus aureus with reduced susceptibility to vancomycin in a patient with erythrodermic psoriasis. AB - A 49-y-old male with erythrodermic psoriasis developed persistent bacteraemia for 3 months due to methicillin-resistant Staphylococcus aureus despite antimicrobial therapy. The skin was the likely focus. Three consecutive isolates from the blood and 1 from the nose were identical and had vancomycin MIC of 4 mg/l. PMID- 17464872 TI - Dysphagia as an unusual presentation of infective endocarditis. AB - We report a case of a 79-y-old male with enterococcal endocarditis presenting with dysphagia and weight loss. The patient's longstanding underlying bacteremia was thought to have caused pro-inflammatory changes leading to alterations in the neuronal environment affecting peripheral nerve function. Dysphagia improved once antibiotic treatment was initiated. PMID- 17464873 TI - An infection with linezolid-resistant S. aureus in a patient with left ventricular assist system. AB - We report an infection with a linezolid-resistant S. aureus in a patient with a left ventricular assist system. Linezolid should be used with caution when invasive devices or foreign materials are in place or therapeutic courses last longer than 14 d. Previous cases of linezolid-resistant S. aureus are summarized. PMID- 17464874 TI - Community-acquired methicillin-resistant Staphylococcus aureus carrying Panton Valentine leukocidin genes: a lethal cause of pneumonia in an adult immunocompetent patient. AB - Necrotizing pneumonia and fatal septic shock were caused by Panton-Valentine leukocidin-positive, community-acquired, methicillin-resistant Staphylococcus aureus (CA-MRSA) in a previously healthy, 61-y-old female. This patient did not belong to any high-risk group (e.g. homosexuals, military recruits, sports team members, etc.). CA-MRSA infection should be suspected in any adult with severe pneumonia/sepsis. PMID- 17464875 TI - Primary sternal osteomyelitis in a healthy child due to community-acquired methicillin-resistant Staphylococcus aureus and literature review. AB - Primary sternal osteomyelitis is a rare condition. Most of the recent cases have been reported in intravenous drug abusers. A 4-y-old male case of primary sternal osteomyelitis due to community-acquired methicillin-resistant Staphylococcus aureus with no apparent risk factors is reported. The diagnosis should be suspected in a young patient presenting with acute inflammatory swelling over the sternum. While bacteriological culture results are pending, antibiotic therapy with Staphylococcus aureus coverage should be initiated empirically and the possibility of community-acquired methicillin-resistant S. aureus must be borne in mind. In this report we also review the literature of paediatric primary sternal osteomyelitis. PMID- 17464876 TI - Anti-ganglioside antibodies-mediated leptospiral meningomyeloencephalopolyneuritis. AB - A case of leptospirosis complicated with meningo-myelo-encephalo-polyneuritis and nephrotic syndrome is presented. Anti-ganglioside antibodies were detected for the first time in a patient with neurological complications of leptospirosis. Possible pathogenic mechanisms and treatment options of these rare manifestations are discussed. PMID- 17464877 TI - Pneumocystis carinii pneumonia in a rheumatoid arthritis patient treated with adalimumab. AB - Patients with rheumatoid arthritis (RA) have an increased risk of infection as a result of alterations in immune regulation, debility, and comorbid illnesses. TNF alpha is of central importance in the pathophysiological responses to infection and inflammation, and plays a crucial role in host defence. Pneumocystis carinii is an opportunistic pathogen that commonly affects individuals with inadequate T cell mediated immune response. Patients with acquired immune deficiency, as well as those receiving immunosuppressive drugs for various conditions have an increased risk of P. carinii pneumonia (PCP). We report the development of PCP in a woman with RA shortly after the initiation of anti-TNF-alpha treatment with adalimumab. PMID- 17464878 TI - Accounting for gene rate heterogeneity in phylogenetic inference. AB - Traditionally, phylogenetic analyses over many genes combine data into a contiguous block. Under this concatenated model, all genes are assumed to evolve at the same rate. However, it is clear that genes evolve at very different rates and that accounting for this rate heterogeneity is important if we are to accurately infer phylogenies from heterogeneous multigene data sets. There remain open questions regarding how best to incorporate gene rate parameters into phylogenetic models and which properties of real data correlate with improved fit over the concatenated model. In this study, two methods of accounting for gene rate heterogeneity are compared: the n-parameter method, which allows for each of the n gene partitions to have a gene rate parameter, and the alpha-parameter method, which fits a distribution to the gene rates. Results demonstrate that the n-parameter method is both computationally faster and in general provides a better fit over the concatenated model than the alpha-parameter method. Furthermore, improved model fit over the concatenated model is highly correlated with the presence of a gene with a slow relative rate of evolution. PMID- 17464879 TI - Profiling phylogenetic informativeness. AB - The resolution of four controversial topics in phylogenetic experimental design hinges upon the informativeness of characters about the historical relationships among taxa. These controversies regard the power of different classes of phylogenetic character, the relative utility of increased taxonomic versus character sampling, the differentiation between lack of phylogenetic signal and a historical rapid radiation, and the design of taxonomically broad phylogenetic studies optimized by taxonomically sparse genome-scale data. Quantification of the informativeness of characters for resolution of phylogenetic hypotheses during specified historical epochs is key to the resolution of these controversies. Here, such a measure of phylogenetic informativeness is formulated. The optimal rate of evolution of a character to resolve a dated four taxon polytomy is derived. By scaling the asymptotic informativeness of a character evolving at a nonoptimal rate by the derived asymptotic optimum, and by normalizing so that net phylogenetic informativeness is equivalent for all rates when integrated across all of history, an informativeness profile across history is derived. Calculation of the informativeness per base pair allows estimation of the cost-effectiveness of character sampling. Calculation of the informativeness per million years allows comparison across historical radiations of the utility of a gene for the inference of rapid adaptive radiation. The theory is applied to profile the phylogenetic informativeness of the genes BRCA1, RAG1, GHR, and c-myc from a muroid rodent sequence data set. Bounded integrations of the phylogenetic profile of these genes over four epochs comprising the diversifications of the muroid rodents, the mammals, the lobe-limbed vertebrates, and the early metazoans demonstrate the differential power of these genes to resolve the branching order among ancestral lineages. This measure of phylogenetic informativeness yields a new kind of information for evaluation of phylogenetic experiments. It conveys the utility of the addition of characters a phylogenetic study and it provides a basis for deciding whether appropriate phylogenetic power has been applied to a polytomy that is proposed to be a rapid radiation. Moreover, it provides a quantitative measure of the capacity of a gene to resolve soft polytomies. PMID- 17464880 TI - Multiple cophylogenetic analyses reveal frequent cospeciation between pelecaniform birds and Pectinopygus lice. AB - Lice in the genus Pectinopygus parasitize a single order of birds (Pelecaniformes). To examine the degree of congruence between the phylogenies of 17 Pectinopygus species and their pelecaniform hosts, sequences from mitochondrial 12S rRNA, 16S rRNA, COI, and nuclear wingless and EF1-alpha genes (2290 nucleotides) and from mitochondrial 12S rRNA, COI, and ATPases 8 and 6 genes (1755 nucleotides) were obtained for the lice and the birds, respectively. Louse data partitions were analyzed for evidence of incongruence and evidence of long-branch attraction prior to cophylogenetic analyses. Host-parasite coevolution was studied by different methods: TreeFitter, TreeMap, ParaFit, likelihood-ratio test, data-based parsimony method, and correlation of coalescence times. All methods agree that there has been extensive cospeciation in this host-parasite system, but the results are sensitive to the selection of different phylogenetic hypotheses and analytical methods for evaluating cospeciation. Perfect congruence between phylogenies is not found in this association, probably as a result of occasional host switching by the lice. Errors due to phylogenetic reconstruction methods, incorrect or incomplete taxon sampling, or to different loci undergoing different evolutionary histories cannot be rejected, thus emphasizing the need for improved cophylogenetic methodologies. PMID- 17464881 TI - Within-species variation and measurement error in phylogenetic comparative methods. AB - Most phylogenetically based statistical methods for the analysis of quantitative or continuously varying phenotypic traits assume that variation within species is absent or at least negligible, which is unrealistic for many traits. Within species variation has several components. Differences among populations of the same species may represent either phylogenetic divergence or direct effects of environmental factors that differ among populations (phenotypic plasticity). Within-population variation also contributes to within-species variation and includes sampling variation, instrument-related error, low repeatability caused by fluctuations in behavioral or physiological state, variation related to age, sex, season, or time of day, and individual variation within such categories. Here we develop techniques for analyzing phylogenetically correlated data to include within-species variation, or "measurement error" as it is often termed in the statistical literature. We derive methods for (i) univariate analyses, including measurement of "phylogenetic signal," (ii) correlation and principal components analysis for multiple traits, (iii) multiple regression, and (iv) inference of "functional relations," such as reduced major axis (RMA) regression. The methods are capable of incorporating measurement error that differs for each data point (mean value for a species or population), but they can be modified for special cases in which less is known about measurement error (e.g., when one is willing to assume something about the ratio of measurement error in two traits). We show that failure to incorporate measurement error can lead to both biased and imprecise (unduly uncertain) parameter estimates. Even previous methods that are thought to account for measurement error, such as conventional RMA regression, can be improved by explicitly incorporating measurement error and phylogenetic correlation. We illustrate these methods with examples and simulations and provide Matlab programs. PMID- 17464882 TI - Geological dates and molecular rates: rapid divergence of rivers and their biotas. AB - We highlight a novel molecular clock calibration system based on geologically dated river reversal and river capture events. Changes in drainage pattern may effect vicariant isolation of freshwater taxa, and thus provide a predictive framework for associated phylogeographic study. As a case in point, New Zealand's Pelorus and Kaituna rivers became geologically isolated from the larger Wairau River system 70 to 130 kyr BP. We conducted mitochondrial DNA phylogeographic analyses of two unrelated freshwater-limited fish taxa native to these river systems (Gobiomorphus breviceps, n = 63; Galaxias divergens, n = 95). Phylogenetic analysis of combined control region and cytochrome b sequences yielded reciprocally monophyletic clades of Pelorus-Kaituna and Wairau haplotypes for each species. Calibrated rates of molecular change based on this freshwater vicariant event are substantially faster than traditionally accepted rates for fishes but consistent with other recent inferences based on geologically young calibration points. A survey of freshwater phylogeographic literature reveals numerous examples in which the ages of recent evolutionary events may have been substantially overestimated through the use of "accepted" calibrations. We recommend that--wherever possible--biologists should start to reassess the conclusions of such studies by using more appropriate molecular calibrations derived from recent geological events. PMID- 17464883 TI - Linking of digital images to phylogenetic data matrices using a morphological ontology. AB - Images are paramount in documentation of morphological data. Production and reproduction costs have traditionally limited how many illustrations taxonomy could afford to publish, and much comparative knowledge continues to be lost as generations turn over. Now digital images are cheaply produced and easily disseminated electronically but pose problems in maintenance, curation, sharing, and use, particularly in long-term data sets involving multiple collaborators and institutions. We propose an efficient linkage of images to phylogenetic data sets via an ontology of morphological terms; an underlying, fine-grained database of specimens, images, and associated metadata; fixation of the meaning of morphological terms (homolog names) by ostensive references to particular taxa; and formalization of images as standard views. The ontology provides the intellectual structure and fundamental design of the relationships and enables intelligent queries to populate phylogenetic data sets with images. The database itself documents primary morphological observations, their vouchers, and associated metadata, rather than the conventional data set cell, and thereby facilitates data maintenance despite character redefinition or specimen reidentification. It minimizes reexamination of specimens, loss of information or data quality, and echoes the data models of web-based repositories for images, specimens, and taxonomic names. Confusion and ambiguity in the meanings of technical morphological terms are reduced by ostensive definitions pointing to features in particular taxa, which may serve as reference for globally unique identifiers of characters. Finally, the concept of standard views (an image illustrating one or more homologs in a specific sex and life stage, in a specific orientation, using a specific device and preparation technique) enables efficient, dynamic linkage of images to the data set and automatic population of matrix cells with images independently of scoring decisions. PMID- 17464884 TI - Foundational issues concerning taxa and taxon names. AB - In a series of articles, Rieppel (2005, Biol. Philos. 20:465-487; 2006a, Cladistics 22:186-197; 2006b, Systematist 26:5-9), Keller et al. (2003, Bot. Rev. 69:93-110), and Nixon and Carpenter (2000, Cladistics 16:298-318) criticize the philosophical foundations of the PhyloCode. They argue that species and higher taxa are not individuals, and they reject the view that taxon names are rigid designators. Furthermore, they charge supporters of the individuality thesis and rigid designator theory with assuming essentialism, committing logical inconsistencies, and offering proposals that render taxonomy untestable. These charges are unsound. Such charges turn on confusions over rigid designator theory and the distinction between kinds and individuals. In addition, Rieppel's, Keller et al.'s, and Nixon and Carpenter's proposed alternatives are no better and have their own problems. The individuality thesis and rigid designator theory should not be quickly abandoned. PMID- 17464885 TI - Distinguishing terminal monophyletic groups from reticulate taxa: performance of phenetic, tree-based, and network procedures. AB - Hybridization is a well-documented, natural phenomenon that is common at low taxonomic levels in the higher plants and other groups. In spite of the obvious potential for gene flow via hybridization to cause reticulation in an evolutionary tree, analytical methods based on a strictly bifurcating model of evolution have frequently been applied to data sets containing taxa known to hybridize in nature. Using simulated data, we evaluated the relative performance of phenetic, tree-based, and network approaches for distinguishing between taxa with known reticulate history and taxa that were true terminal monophyletic groups. In all methods examined, type I error (the erroneous rejection of the null hypothesis that a taxon of interest is not monophyletic) was likely during the early stages of introgressive hybridization. We used the gradual erosion of type I error with continued gene flow as a metric for assessing relative performance. Bifurcating tree-based methods performed poorly, with highly supported, incorrect topologies appearing during some phases of the simulation. Based on our model, we estimate that many thousands of gene flow events may be required in natural systems before reticulate taxa will be reliably detected using tree-based methods of phylogeny reconstruction. We conclude that the use of standard bifurcating tree-based methods to identify terminal monophyletic groups for the purposes of defining or delimiting phylogenetic species, or for prioritizing populations for conservation purposes, is difficult to justify when gene flow between sampled taxa is possible. As an alternative, we explored the use of two network methods. Minimum spanning networks performed worse than most tree-based methods and did not yield topologies that were easily interpretable as phylogenies. The performance of NeighborNet was comparable to parsimony bootstrap analysis. NeighborNet and reverse successive weighting were capable of identifying an ephemeral signature of reticulate evolution during the early stages of introgression by revealing conflicting phylogenetic signal. However, when gene flow was topologically complex, the conflicting phylogenetic signal revealed by these methods resulted in a high probability of type II error (inferring that a monophyletic taxon has a reticulate history). Lastly, we present a novel application of an existing nonparametric clustering procedure that, when used against a density landscape derived from principal coordinate data, showed superior performance to the tree-based and network procedures tested. PMID- 17464886 TI - Genomic analysis and geographic visualization of the spread of avian influenza (H5N1). PMID- 17464887 TI - Properties of supertree methods in the consensus setting. PMID- 17464888 TI - Alarm bells for the molecular clock? No support for Ho et al.'s model of time dependent molecular rate estimates. PMID- 17464889 TI - Seeing the trees for the network: consensus, information content, and superphylogenies. PMID- 17464890 TI - How many genes should a systematist sample? Conflicting insights from a phylogenomic matrix characterized by replicated incongruence. PMID- 17464894 TI - [Guideline: treatment of adult obstructive sleep apnea]. PMID- 17464895 TI - [The effectiveness of an individual mandibular advancement splint (MAS) in subjects with obstructive tongue-base-snoring]. AB - OBJECTIVE: Besides operation the mandibular advancement splint (MAS) is one of the main options for treatment of snoring. Especially the MAS by Hinz, which exclusively blocks mandibular retrusion, has been established as an effective alternative. The aim of this study is to examine the effectiveness of this MAS and to evaluate the role of obstruction at the level of the tongue base as a successful pretherapeutic predictor by using the sleep nasal endoscopy. SUBJECTS AND METHODS: From August 2001 until August 2006 we have examined over 450 patients with sleep disorders at our hospital. All patients have had an ENT examination, a standardized polysomnography and a sleep nasal endoscopy with therapy simulation. The application of a MAS is indicated if an obstruction at the level of the tongue base is detected and the RDI (respiratory disturbance index) is below 10/h. 10 patients are examined by PSG and partly by sleep nasal endoscopy before and after adjustment of the MAS. The patient group is classified in three velum- (n = 3), isolated tongue base-snorers, five combined tongue base /velum- and two tongue base-/epiglottis-snorers. RESULTS: The subjective sleep recovery and the snoring-intensity can be improved by the MAS in eight of ten patients. In addition the long-term acceptance of this therapy is very high. CONCLUSIONS: According to our experiences the MAS by Hinz is an effective treatment option for patients with an obstruction level at the tongue base confirmed by sleep nasal endoscopy. PMID- 17464896 TI - [Chance or causality: problems recognizing laryngeal carcinoma as a result of occupational exposition to noxious substances of blue collar worker employed in the rubber industry]. AB - BACKGROUND: Laryngeal cancer as an occupational disease in the rubber industry is under subject of continuous epidemiological research because of the lack of stable statistical data. METHOD: Epidemiological studies published in the period between 1982 and 2006 were analysed in regard of the risks employment and laryngeal carcinoma. 23 own cases of medical expert opinion and the documentation of the German Occupational Cooperatives were analysed. RESULTS: A significant statistical correlation between a specific exposition to cancerogenics in the rubber industry and a development of laryngeal carcinoma could not be found in the literature, still there is evidence for an increased mortality. It points to asbestos, talcum, dust as one risk factor. CONCLUSION: Recognition and compensation for laryngeal carcinoma according to [section sign] 9 Abs. 2 SGB VII is possible after intense individual investigation, although up to now no clear statistical evidence for the coincidence between employment in the rubber industry and general could be proven. PMID- 17464897 TI - [Crossed aphasia or dysexecutive syndrome? A case report]. AB - The case of a female dextral patient (MN) is reported. MN sustained a partial frontotemporal infarction in the right hemisphere. Clinically, MN presented herself mainly with aphasic symptoms. The dextrality of the patient and the laterality of the lesion as well as the nature of language deficits would legitimate the diagnosis of a crossed aphasia, implying paradoxical lateralization of language in MN. However, comprehensive neuropsychological assessment revealed the presence of a global dysexecutive syndrome in MN, suggesting a general cognitive impairment, part of which must be the aphasic symptoms. General issues regarding the relationship between language and cognition and the lateralization of cognitive functions are discussed. PMID- 17464898 TI - [Non-cognitive symptoms and psychopharmacological treatment in demented nursing home residents]. AB - Dementias, in particular Alzheimer's disease (AD), are the main reason for availing of nursing home care. In the course of the illness, the clinical picture is affected by cognitive decline and by other psychopathological, "non-cognitive" symptoms such as apathy, depression, delusions or agitation. Little attention has been paid to these symptoms, although they lead to an increase in strain on the patients and their relatives as well as complications in nursing care. Psychopathological symptoms were evaluated by using the Neuropsychiatric Inventory in 145 nursing home residents (age: 85 +/- 7 years, duration of stay: 35 +/- 48 months); the majority of them with moderate to severe dementia (GDS: 5 +/- 2; MMSE: 11 +/- 9). In addition, the Apathy Evaluation Scale was applied. To meet potential regional effects, residents were recruited in nursing homes in the areas around Heidelberg as well as Munster. 87% of the participants showed psychopathological symptoms of an at least moderate degree, depressive mood (52%), apathy (41%) and agitation (38%) being most frequent. General condition, nutritional status and care status were evaluated as 'good', likewise general health care. In contrast, only 27% were treated by psychiatrists. 70% received psychopharmacological treatment, mostly sedatives (44%), while antidementive drugs were used only in 11%. The findings underline the need of further information and advanced training. PMID- 17464901 TI - [Current teaching, learning and examination methods in medical education and potential applications in rehabilitative issues]. AB - With introduction of the new Federal Medical Licensing Regulations (Approbationsordnung) in Germany, integrated teaching in "Rehabilitation, Physical Medicine, Naturopathic Treatment" (Querschnittsbereich Q12) has become obligatory for the first time. Furthermore, the new Regulations require the medical faculties in Germany to realize an innovative didactic orientation in teaching. This paper provides an overview of recent applications of teaching techniques and examination methods in medical education with special consideration of the new integrated course Q12 and further teaching methods related to rehabilitative issues. Problem-oriented learning (POL), problem-based learning (PBL), bedside teaching, eLearning, and the examination methods Objective Structured Clinical Examination (OSCE) and Triple Jump are in the focus. This overview is intended as the basis for subsequent publications of the Commission for Undergraduate and Postgraduate Training of the German Society of Rehabilitation Science (DGRW), which will present examples of innovative teaching material. PMID- 17464902 TI - [Rehabilitation and Diagnosis Related Groups (REDIA Study): impact of DRG introduction in the acute sector on medical rehabilitation in Germany]. AB - As experiences from other countries show, introduction and use of Diagnosis Related Groups (DRG), as of January 2004 now also mandatory in Germany, may have a significant impact on associated rehabilitation. The Institute of Hospital Management (IKM) in a multi-centre study promoted by Deutsche Rentenversicherung Bund and Deutsche Rentenversicherung Westfalen is conducting a study regarding potential diversion of healthcare expenditures from acute care towards rehabilitation as a result of DRG introduction in Germany. For documentation of potential short-term changes in patient populations and patient streams, extensive data have been collected in the first two phases in 2003/04 and 2005/06 for a total of 1342 cardiologic and orthopaedic patients. Indication-specific comparison of the two phases showed significantly shorter stays in the acute sector as well as shorter transition times between the sectors, resulting in an intake of patients into rehabilitative care at an earlier stage of their recovery process. Significant diversion of treatment efforts from the acute to the rehabilitative sector, regarding increased nursing effort and potential changes in the therapeutic and medical treatment to be provided, has not been proven as yet. The increase in wound problems expected by practitioners was confirmed in the orthopaedic area by an increasing number of wound healing disturbances and haematomas; in bypass-patients, an increasing number of pericardium and pleura bruises was found. The analyses performed on the data collected revealed no limitations in the patients' ability to participate in rehabilitative measures when the first and the second phase of the study are compared. To be able to depict the further course and interdependencies of changes, continuous systematic observation of developments would be desirable. To ascertain a lasting impact of DRG implementation at least a third study-phase will be necessary, which should be placed at the end of 2008, at the time when the DRG convergence phase will end. PMID- 17464903 TI - [Diagnostic instruments for occupational stress experience and coping in psychosomatic rehabilitation]. AB - BACKGROUND: Psychosomatic rehabilitation research increasingly agrees that symptom-related therapies need to be supplemented by work-related therapeutic interventions. However, there is a lack of evaluated diagnostic instruments for determination of an indication for and evaluation of specific work-related interventions. This article presents an analysis of the diagnostic quality of work-related self-rating instruments as well as their evaluation from a practical perspective. METHOD: 281 psychosomatic inpatients showing high levels of occupational stress were studied using a set of work-related questionnaires on admission: individual coping skills (AVEM), work-related therapy motivation (FBTM), work satisfaction (ABB), leisure activities, experienced control at work, as well as occupational concerns and social stressors at work. Furthermore a sub group of 78 patients who had completed a specific work-related group programme during their inpatient treatment were interviewed three months post-treatment. The instruments used were examined for reliability, underlying dimensions (factor analysis), health relevance (correlations with the SCL-90-R total score GSI as health-related external criterion), and sensitivity to change. RESULTS: (1) Reliability: Measuring procedures fulfilled the requirements of measurement accuracy. (2) Underlying dimensions: Factor-analytical investigations of relevant occupational constructs identified six aspects of occupational coping relevant for rehabilitation practice: "occupational resilience", "occupational (over )commitment", "occupational gratification", "well-being at work", a motivational factor of "occupational identification vs. withdrawal", and "leisure activities". (3) Health relevance: Variables of "occupational resilience" and "occupational gratification" showed the highest correlations with indicators of psychosomatic health. (4) Sensitivity to change: Work-related instruments proved to be predominantly sensitive to change. CONCLUSIONS: The work-related self-report instruments proved satisfactorily in application with psychosomatic patients. The findings concerning their diagnostic quality show that they can be recommended in research to evaluate work-related issues. Longitudinal evaluations should include work-related diagnostic instruments to a much larger extent. PMID- 17464904 TI - [Neuropsychology in occupational rehabilitation: a new field of intervention?]. AB - The main goal of rehabilitation is social and vocational reintegration of handicapped people. Neuropsychological disorders prove to be particularly critical factors in this context. In many cases vocational retraining requiring good cognitive abilities and capacity is essential. Neuropsychological impairments can substantially diminish the chance of successful vocational rehabilitation. To assess the need for neuropsychological intervention within the scope of vocational rehabilitation programmes, 69 persons undergoing rehabilitation in the Berufsforderungswerk Sachsen-Anhalt, a non-profit organization devoted to vocational reintegration, were screened to identify individuals with cognitive deficits. Furthermore a brief multidimensional self report inventory, the Symptom Checklist-90-R (SCL-90-R) was administered. In addition, the files of all participants were reviewed with regard to mental or neurological disorders. The study revealed cognitive deficits in a quarter to a third of the participants depending on the cognitive domain. Comparison of neuropsychological test performance of trainees with and without established diagnosis of pertinent diseases confirmed cognitive deficits in those with prior diagnosis of neurological disorder, as expected. Only for Divided Attention a significant correlation was found between neuropsychological deficits and psychiatric disorders as documented in the files. However, a considerable number of persons without corresponding medical-psychological pre-diagnosis were found to also perform poorly in particular tests. Furthermore it could be shown that subjectively experienced stress in trainees had a negative impact on reaction times in specific test procedures. These findings strongly suggest a need to supplement vocational rehabilitation by neuropsychological training programmes. When cognitive functional impairments are suspected, specific diagnostic assessments should be administered in order to select and implement appropriate interventions. PMID- 17464905 TI - [A coping with Stress Training (SBT) for persons with mental illness--pilot study on a group training programme in support of occupational rehabilitation]. AB - OBJECTIVE: This pilot study investigates the effects of a coping with stress training (SBT) programme which had been offered in the Workshop for Disabled People of the Stuttgart Occupational Rehabilitation Center Rudolf-Sophien-Stift to persons with mental illness. The group training is aimed at supporting the participants in coping effectively with stress, avoiding over- or under stimulation and strengthening positive experiences. METHOD: In a control-group study design, data was collected from 19 participants of the training group and 11 participants of a (waiting) control group. Data collection was carried out before and after the training. RESULTS: In the pre-post comparison the training group shows a reduction of symptoms in general and especially in somatization, obsessive-compulsiveness, anxiety, and paranoid ideation. The participants showed improvements in quality of life in the areas of capability, ability to enjoy and relax, positive and negative mood. In respect of attitudes of competence and control they showed improvements in internal control attitudes. Also, as regards illness coping and prevalent coping styles, active and problem-oriented coping styles were found to have increased to some extent. In the problem solving training, 84.2% of the participants stated in their self-ratings that they had been able to achieve positive changes in a specific problem area. CONCLUSIONS: From a methodological angle it has to be pointed out that the study had examined a very small group. Although a number of open questions remain the results obtained are encouraging and in accordance with the objectives of the training. Further studies need to be carried out with larger samples from different facilities in the fields of medical, vocational and social rehabilitation. PMID- 17464906 TI - [The legal relationship between social authorities and the rehabilitation services and institutions]. PMID- 17464907 TI - [Rehabilitation of young people with chronic illness--current situation and outlook. Report of the Concluding Conference of a Project of the Federal Ministry of Labour and Social Issues, Oct. 13-14, 2005 in Wurzburg]. PMID- 17464908 TI - Risk of rabies infection and adverse effects of postexposure prophylaxis in healthcare workers and other patient contacts exposed to a rabies virus-infected lung transplant recipient. AB - BACKGROUND: Rabies virus was inadvertently transmitted to a lung transplant recipient through donor lungs. The patient was given ventilatory assistance and cared for postoperatively for 6 weeks before a diagnosis of rabies virus infection was made. Postexposure prophylaxis (PEP) was offered to potentially exposed healthcare workers (HCWs). METHODS: Only HCWs classified as belonging to possible and/or proven contact groups (according to a standardized interview) received PEP. The risk of individual HCWs being exposed to rabies virus was reassessed on the basis of viral concentrations measured in the patient's excretions and body fluids. HCWs who were vaccinated as part of PEP were followed up prospectively according to a standardized procedure. RESULTS: Of 179 HCWs and other patient contacts, 132 met the eligibility criteria for PEP (118 [89.4%] with possible contact and 14 [10.6%] with proven contact with the patient's excretions and/or body fluids). One hundred thirty-one individuals started PEP, and 126 met the inclusion criteria for analysis. Of these, 48 (38%) developed at least 1 adverse effect (8 [6.3%] had fever, 37 [29.4%] had headache, 3 [2.4%] had lymphadenopathy, 17 [13.5%] had dizziness, and 6 [4.8%] had paresthesia). No HCW or other patient contact developed rabies or serious PEP-related adverse effects. Reassessment of the individual's risk of infection as a function of the viral concentration in the patient's excretions and/or body fluids (up to 5.12 x 10(7) copies/mL) revealed that 103 HCWs (78.0%) had contact with high-risk substances (89 [67.40%] had possible contact and 14 [10.7%] had proven contact). CONCLUSION: HCWs can be exposed to significant viral concentrations in excretions and/or body fluids from rabies virus-infected lung transplant recipients. Because widespread use of PEP entails the possibility of significant health problems for HCWs considered to be at risk of contracting rabies, applying a rational indication for PEP is crucial. PMID- 17464909 TI - Infectivity of hepatitis C virus in plasma after drying and storing at room temperature. AB - OBJECTIVE: To determine effect of environmental exposure on the survival and infectivity of hepatitis C virus (HCV). METHODS: Three aliquots of chimpanzee plasma containing HCV and proven infectious HCV inoculum were dried and stored at room temperature, 1 aliquot for 16 hours, 1 for 4 days, and 1 for 7 days. A chimpanzee (CH247) was sequentially inoculated intravenously with each of these experimental inocula, beginning with the material stored for 7 days. Each inoculation was separated by at least 18 weeks of follow-up to monitor for infection. The concentration of HCV RNA was measured and quasi species were sequenced for each experimental inoculum and in serum samples from CH247. RESULTS: Evidence of HCV infection developed in CH247 only after inoculation with the material stored for 16 hours. No infection occurred after inoculation with the material stored for 7 days or 4 days. Compared with the original infectious chimpanzee plasma, the concentration of HCV RNA was 1 log lower in all 3 experimental inocula. The same predominant sequences were found in similar proportions in the original chimpanzee plasma and in the experimental inocula, as well as in serum samples from CH247. CONCLUSION: HCV in plasma can survive drying and environmental exposure to room temperature for at least 16 hours, which supports the results of recent epidemiologic investigations that implicated blood contaminated inanimate surfaces, objects, and/or devices as reservoirs for patient-to-patient transmission of HCV. Healthcare professionals in all settings should review their aseptic techniques and infection control practices to ensure that they are being performed in a manner that prevents cross-contamination from such reservoirs. PMID- 17464910 TI - Assessment of intervention measures for the 2003 SARS epidemic in Taiwan by use of a back-projection method. AB - OBJECTIVES: To reconstruct the infection curve for the 2003 severe acute respiratory syndrome (SARS) epidemic in Taiwan and to ascertain the temporal changes in the daily number of infections that occurred during the course of the outbreak. METHOD: Back-projection method. RESULTS: The peaks of the epidemic correspond well with the occurrence of major infection clusters in the hospitals. The overall downward trend of the infection curve after early May corresponds well to the date (May 10) when changes in the review and classification procedure were implemented by the SARS Prevention and Extrication Committee. CONCLUSION: The major infection control measures taken by the Taiwanese government over the course of the SARS epidemic, particularly those regarding infection control in hospitals, played a crucial role in containing the outbreak. PMID- 17464911 TI - Policies for endotracheal suctioning of patients receiving mechanical ventilation: a systematic review of randomized controlled trials. AB - OBJECTIVE: The Dutch Working Party on Infection Prevention (Werkgroep Infectiepreventie [WIP]) aimed to determine whether certain policies on endotracheal suctioning are better than others in terms of prevention of ventilator-associated pneumonia (VAP) in patients receiving mechanical ventilation in the intensive care unit. METHODS: Publications were retrieved by a systematic search of Medline and the Cochrane Library for literature published before February 2006. Additionally, the reference lists of all identified trials were examined. All randomized trials, quasi-randomized trials, and systematic reviews or meta-analyses of randomized or quasi-randomized trials that compared different policies on endotracheal suctioning for patients receiving mechanical ventilation in the intensive care unit were selected. Two reviewers independently assessed trial quality and extracted data. Disagreements were resolved by discussion with a third reviewer. Data from the original publications were used to calculate the relative risk of VAP. Data for VAP were combined in the analysis where appropriate, by use of a random-effects model. RESULTS: Ten trials were included in the review. The quality of the trials and the way they were reported were generally unsatisfactory. Eight low-quality trials indicate that use of closed instead of open suction systems has no effect on the incidence of VAP. Two moderate-quality trials indicate that changing in-line suction catheters less frequently than every 24 hours does not increase the incidence of VAP. CONCLUSION: The WIP recommends that there be no preferential use of either open or closed endotracheal suction systems to reduce the rate of VAP, but it elucidates that the quality of the evidence is low. Considerations other than prevention of VAP should determine the choice of the suction system. When closed systems are used, the WIP recommends changing the in-line suction catheters every 48 hours. In case of mechanical failure or soiling of the suction system, they may be changed more frequently. PMID- 17464912 TI - Two nosocomial pertussis outbreaks and their associated costs - King County, Washington, 2004. AB - OBJECTIVE: Pertussis outbreaks in healthcare settings result in resource intensive control activities, but studies have rarely evaluated the associated costs. We describe and estimate costs associated with 2 nosocomial pertussis outbreaks in King County, Washington, during the period from July 25 to September 15, 2004. One outbreak occurred at a 500-bed tertiary care hospital (hospital A), and the other occurred at a 250-bed pediatric hospital (hospital B). METHODS: We estimated the costs of each outbreak from the hospitals' perspective through standardized interviews with hospital staff and review of contact tracing logs. Direct costs included personnel time and laboratory and medication costs, whereas indirect costs were those resulting from hospital staff furloughs. RESULTS: Hospital A incurred direct costs of $195,342 and indirect costs of $68,015; hospital B incurred direct costs of $71,130 and indirect costs of $50,000. Cost differences resulted primarily from higher personnel costs at hospital A ($134,536), compared with hospital B ($21,645). Total cost per pertussis case was $43,893 for hospital A (6 cases) and $30,282 for hospital B (4 cases). Total cost per person exposed to a pertussis patient were $357 for hospital A (738 exposures) and $164 for hospital B (737 exposures). CONCLUSIONS: Nosocomial pertussis outbreaks result in substantial costs to hospitals, even when the number of pertussis cases is low. The cost-effectiveness of strategies to prevent nosocomial pertussis outbreaks, including vaccination of healthcare workers, should be evaluated. PMID- 17464913 TI - Circumstances of patient falls and injuries in 9 hospitals in a midwestern healthcare system. AB - OBJECTIVE: Preventing hospital falls and injuries requires knowledge of fall and injury circumstances. Our objectives were to determine whether reported fall circumstances differ among hospitals and to identify predictors of fall-related injury. DESIGN: Retrospective cohort study. Adverse event data on falls were compared according to hospital characteristics. Logistic regression was used to determine adjusted odds ratios (aORs) with 95% confidence intervals (CIs) for risk factors for fall-related injury. SETTING: Nine hospitals in a Midwestern healthcare system. PATIENTS: Inpatients who fell during 2001-2003. RESULTS: The 9 hospitals reported 8,974 falls that occurred in patient care areas, involving 7,082 patients; 7,082 falls were included in our analysis. Assisted falls (which accounted for 13.3% of falls in the academic hospital and 9.8% of falls in the nonacademic hospitals; P<.001) and serious fall-related injuries (which accounted for 3.7% of fall-related injuries in the academic hospital and 2.2% of fall related injuries in the nonacademic hospitals; P<.001) differed by hospital type. In multivariate analysis for the academic hospital, increased age (aOR, 1.006 [95% CI, 1.000-1.012]), falls in locations other than patient rooms (aOR, 1.53 [95% CI, 1.03-2.27]), and unassisted falls (aOR, 1.70 [95% CI, 1.23-2.36]) were associated with increased injury risk. Altered mental status was associated with a decreased injury risk (aOR, 0.72 [95% CI, 0.58-0.89]). In multivariate analysis for the nonacademic hospitals, increased age (aOR, 1.007 [95% CI, 1.002-1.013]), falls in the bathroom (aOR, 1.46 [95% CI, 1.06-2.01]), and unassisted falls (aOR, 1.83 [95% CI, 1.37-2.43]) were associated with injury. Female sex (aOR, 0.83 [95% CI, 0.71-0.97]) was associated with a decreased risk of injury. CONCLUSION: Some fall characteristics differed by hospital type. Further research is necessary to determine whether differences reflect true differences or merely differences in reporting practices. Fall prevention programs should target falls involving older patients, unassisted falls, and falls that occur in the patient's bathroom and in patient care areas outside of the patient's room to reduce injuries. PMID- 17464914 TI - Evaluation of antimicrobial therapy orders circumventing an antimicrobial stewardship program: investigating the strategy of "stealth dosing". AB - OBJECTIVE: Prior-approval antimicrobial stewardship programs (ASPs) improve patient outcomes and decrease antimicrobial resistance. These benefits would be limited if physicians circumvented ASP efforts. We evaluated whether prescribers wait until after the prior-approval period to order restricted antimicrobial therapy that is in conflict with guidelines or unnecessary. DESIGN: A cross sectional study design and a retrospective cohort study design. SETTING: A tertiary care, academic medical center with a prior-approval ASP that was active between 8 am and 10 pm. METHODS: We evaluated whether there was an increase in the proportion of orders for antimicrobial therapy that involve restricted (vs nonrestricted) antimicrobials during the first hour that the ASP is inactive (ie, the first hour that prior approval is not required), compared with the remainder of the day. We also evaluated whether restricted antimicrobial therapy ordered during this first hour is less likely to be continued when the ASP becomes active the next day, compared with that ordered during the preceding hour. RESULTS: A greater proportion of the antimicrobial therapy orders placed between 10:00 pm and 10:59 pm were for restricted agents, compared with orders placed during other periods (57.0% vs 49.9%; P=.02). Surgical patients for whom antimicrobial therapy orders were placed between 10:00 pm and 10:59 pm were less likely to have that antimicrobial therapy continued, compared with patients whose therapy was ordered between 9:00 pm and 9:59 pm (60.0% vs 98.1%; P<.001). Nonsurgical patients whose therapy orders were placed between 10:00 pm and 10:59 pm were also less likely to have the ordered antimicrobial therapy continued, compared with patients whose therapy was ordered between 9:00 pm and 9:59 pm (70.8% vs 84.2%; P=.01). CONCLUSION: Physicians avoid having to obtain prior approval for therapy involving restricted antimicrobials by waiting until restrictions are no longer active to place orders. Compared with restricted antimicrobial therapy ordered when the ASP is active, these courses of therapy are less often continued by the ASP, suggesting that they are more likely to be in conflict with guidelines or unnecessary. PMID- 17464915 TI - Associations between surgical site infection risk and hospital operation volume and surgeon operation volume among hospitals in the Dutch nosocomial infection surveillance network. AB - OBJECTIVE: To examine the association between hospital operation volume and surgeon operation volume and the risk of surgical site infection (SSI). DESIGN: Prospective, multicenter cohort study based on surveillance data. METHODS: Data were obtained from the Dutch surveillance network for nosocomial infections (Preventie Ziekenhuisinfecties door Surveillance [PREZIES]) on 9 different types of orthopedic surgery, general surgery, and gynecology procedures performed during 1996-2003. Multilevel logistic regression analysis was performed to assess the independent effect of hospital volume and surgeon volume on SSI risk. RESULTS: Hospital volume was not significantly associated with SSI risk for any of the selected procedures. Low surgeon volume was associated with an increased risk for an infection for 7 of 9 types of procedures, although this effect was statistically significant only for knee arthroplasty. For 4 procedures, the odds of exceeding the 75th percentile for duration of surgery were greater when the surgeon volume was low than when the surgeon volume was moderate or high. CONCLUSIONS: Patients operated on by surgeons with a low operation volume seem to have a higher risk of developing an SSI with some procedures, particularly knee arthroplasty. The higher SSI risk for surgeons with a low operation volume is possibly partly mediated by the longer duration of surgery, a well-known risk factor for development of SSI. PMID- 17464916 TI - Immunity against infectious diseases: predictive value of self-reported history of vaccination and disease. AB - OBJECTIVE: To determine whether self-reported history of disease and/or vaccination is predictive of immunity against hepatitis B, varicella, rubella, mumps, and measles. DESIGN: The seroprevalence of viral antibodies and the predictive value of a self-report questionnaire were determined for 616 paramedical students who matriculated into Padua Medical School (Padua, Italy) during 2003-2005. RESULTS: The majority of subjects (86.9%) remembered being vaccinated against hepatitis B but had no recollection of disease. Among vaccinees, 1.5% showed markers of previous infection, 6.7% tested negative for anti-hepatitis B virus surface antigen (anti-HBsAg) antibodies, and 91.8% tested positive for anti-HBsAg. Self-reported vaccination history had a positive predictive value of 93.2% for test results positive for immunity against hepatitis B. Immunity against varicella (93.7% of subjects) and rubella (95.5%) was high, compared with immunity against mumps (79.9%) and measles (83.1%). In addition, results of tests for detection of immunity against mumps and measles were equivocal for more than 7% of subjects, probably because their vaccination regimen was not completed. Self-reported histories of varicella disease and rubella disease and vaccination had high positive predictive values (greater than 98% each) for testing positive for antiviral antibodies, compared with self reported histories of mumps disease and vaccination and measles disease and vaccination; however, high positive predictive values were observed for self reported histories of mumps only (92.0%) and measles only (94.7%). CONCLUSIONS: The self-report questionnaire used in this study did not accurately predict immunity against 5 transmittable but vaccine-preventable diseases. A complete serological evaluation of healthcare workers, followed by vaccination of those with negative or equivocal results of serological tests, is an appropriate measure to decrease the risk of infection in this population. PMID- 17464917 TI - Prospective observational study of candidemia in Sao Paulo, Brazil: incidence rate, epidemiology, and predictors of mortality. AB - BACKGROUND: Studies conducted in tertiary care hospitals of different European countries and the United States have shown incidence rates of candidemia ranging from 0.17 to 0.76 and 0.28 to 0.96 per 1,000 admissions, respectively. So far, only 1 study has evaluated the incidence rates of candidemia in tertiary care hospitals in Latin American countries. OBJECTIVE: To evaluate the epidemiology of candidemia in 4 tertiary care hospitals in Sao Paulo, Brazil. DESIGN: Multicenter, laboratory-based surveillance of candidemia. RESULTS: A total of 7,038 episodes of bloodstream infection were identified, and Candida species accounted for 282 cases (4%). The incidence rate of candidemia was 1.66 candidemic episodes per 1,000 hospital admissions. Candida albicans was the most frequently isolated Candida species in all hospitals, but Candida species other than C. albicans accounted for 62% of isolates, including predominantly Candida parapsilosis and Candida tropicalis. Azole resistance was restricted to only 2% of all Candida isolates (1 isolate of Candida glabrata and 4 isolates of Candida rugosa). Candidemia was mostly documented in surgical patients with long durations of hospital stay. The crude mortality rate was 61%, and advanced age and high Acute Physiology and Chronic Health Evaluation II score were both conditions independently associated with risk of death. CONCLUSIONS: We observed in our series a higher incidence rate of candidemia than that reported in European countries and the United States. Advanced age and a high Acute Physiology and Chronic Health Evaluation II score were factors associated with a higher probability of death in candidemic patients. Fluconazole-resistant Candida strains are still a rare finding in our case-based study of candidemia. PMID- 17464918 TI - Effect of a chlorhexidine mouthwash on the risk of postextraction bacteremia. AB - OBJECTIVE: To investigate the prevalence, duration, and etiology of bacteremia following dental extractions performed after a single administration of chlorhexidine mouthwash. DESIGN AND SETTING: A randomized, controlled trial performed in a university hospital. METHODS: A series of 106 patients with mental and behavioral disabilities who underwent dental extractions under general anesthesia were randomly assigned to a control group or chlorhexidine group. The exclusion criteria applied were use of antibiotics in the previous 3 months, use of oral antiseptics, any type of congenital or acquired immunodeficiency, and disease that predisposes the patient to infections or bleeding. The chlorhexidine group had 0.2% chlorhexidine mouthwash administered for 30 seconds before any dental manipulation. Blood samples were collected at baseline, 30 seconds, 15 minutes, and 1 hour after the dental extractions. Subculture and further identification of the isolated bacteria were performed by conventional microbiological techniques. RESULTS: The prevalence of bacteremia after dental extraction in the control and chlorhexidine groups were 96% and 79%, respectively, at 30 seconds (P=.008), 64% and 30% at 15 minutes (P<.001), and 20% and 2% at 1 hour (P=.005). The most frequently identified bacteria were Streptococcus species in both the control and chlorhexidine groups (64% and 68%, respectively), particularly viridans group streptococci. CONCLUSION: We recommend the routine use of a 0.2% chlorhexidine mouthwash before dental extractions to reduce the risk of postextraction bacteremia. PMID- 17464919 TI - Reduction in surgical site infections in neurosurgical patients associated with a bedside hand hygiene program in Vietnam. AB - OBJECTIVE: We conducted an intervention study to assess the impact of the use of an alcohol-chlorhexidine-based hand sanitizer on surgical site infection (SSI) rates among neurosurgical patients in Ho Chi Minh City, Vietnam. DESIGN: A quasi experimental study with an untreated control group and assessment of neurosurgical patients admitted to 2 neurosurgical wards at Cho Ray Hospital between July 11 and August 15, 2000 (before the intervention), and July 14 and August 18, 2001 (after the intervention). A hand sanitizer with 70% isopropyl alcohol and 0.5% chlorhexidine gluconate was introduced, and healthcare workers were trained in its use on ward A in September 2000. No intervention was made in ward B. Centers for Disease Control and Prevention definitions of SSI were used. Patient SSI data were collected on standardized forms and were analyzed using Stata software (Stata). RESULTS: A total of 786 patients were enrolled: 377 in the period before intervention (156 in ward A and 221 in ward B) and 409 in the period after intervention (159 in ward A and 250 in ward B). On ward A after the intervention, the SSI rate was reduced by 54% (from 8.3% to 3.8%; P=.09), and more than half of superficial SSIs were eliminated (7 of 13 vs 0 of 6 in ward B; P=.007). On ward B, the SSI rate increased by 22% (from 7.2% to 9.2%; P=.8). In patients without SSI, the median postoperative length of stay and the duration of antimicrobial use were reduced on ward A (both from 8 to 6 days; P<.001) but not on ward B. CONCLUSIONS: Our study demonstrates that introduction of a hand sanitizer can both reduce SSI rates in neurosurgical patients, with particular impact on superficial SSIs, and reduce the overall postoperative length of stay and the duration of antimicrobial use. Hand hygiene programs in developing countries are likely to reduce SSI rates and improve patient outcomes. PMID- 17464920 TI - Political versus epidemiological correctness. PMID- 17464921 TI - Politically incorrect: legislation must not mandate specific healthcare epidemiology and infection prevention and control practices. PMID- 17464922 TI - Efficacy of a silicone urinary catheter impregnated with chlorhexidine and triclosan against colonization with Proteus mirabilis and other uropathogens. AB - We sought to develop an infection-resistant urinary catheter. We evaluated 3 types of catheters for their efficacy against uropathogens in an in vitro model of the urinary tract. The catheter impregnated with chlorhexidine and triclosan suppressed the growth of uropathogens, including Proteus mirabilis, for 20-30 days or longer. PMID- 17464923 TI - Central venous catheter-related Streptomyces septic thrombosis. AB - Streptomyces species are part of the actinomycetes group. They have rarely been reported as a cause of invasive infection. We report a case of catheter-related Streptomyces bacteremia complicated by severe sepsis and septic thrombosis. We also present a brief review of the literature on Streptomyces bacteremia. PMID- 17464924 TI - Device-associated nosocomial infection rates in intensive care units in Greece. AB - Site-specific, risk-adjusted incidence rates of intensive care unit (ICU) acquired infections were obtained through standardized surveillance in 8 ICUs in Greece. High rates were observed for central line-associated bloodstream infection (12.1 infections per 1,000 device-days) and ventilator-associated pneumonia (12.5 infections per 1,000 device-days). Gram-negative microorganisms accounted for 60.4% of the isolates recovered, and Acinetobacter species were predominant. To reduce infection rates in Greek ICUs, comprehensive infection control programs are required. PMID- 17464925 TI - Healthcare costs associated with hemodialysis catheter-related infections: a single-center experience. AB - In patients undergoing hemodialysis, catheter-related bacteremia results in expensive hospitalizations. In our study, the mean cost was $23,451 per hospitalization. When itemized, housing ("bed-related") costs accounted for 66% of the total; laboratory costs accounted for 4%, radiologic costs accounted for 9%, and procedure-related costs accounted for 21%. Hypoalbuminemia and bacteremia due to methicillin-resistant Staphylococcus aureus (MRSA) are associated with higher healthcare costs; bacteremia due to MRSA is also associated with poor survival rates. PMID- 17464926 TI - Incidence of catheter-related bloodstream infection among patients with a needleless, mechanical valve-based intravenous connector in an Australian hematology-oncology unit. AB - There are few Australian data on the incidence of catheter-associated bloodstream infection (BSI) among patients in hematology-oncology units. We found an increase in catheter-associated BSI rates coincident with the introduction of a mechanical valve connector (2.6 infections vs 5.8 infections per 1,000 catheter-days; incidence rate ratio, 2.2; P=.031). PMID- 17464927 TI - Streptococcal meningitis following myelogram procedures. AB - In September of 2004, we investigated 7 cases of post-myelography meningitis. Streptococcal species were recovered from blood or cerebrospinal fluid in all cases. Our findings suggest that droplet transmission of the oral flora of the clinician performing the procedure was the most likely source of these infections. The Centers for Disease Control and Prevention recommends the use of face masks by those performing myelograms. PMID- 17464928 TI - Hospital "self-prophylaxis": strategies for efficient protection of the workforce in the face of infectious disease threats. AB - Hospital preparedness for nosocomial or community-wide outbreaks of communicable disease includes the capability for rapid, self-reliant administration of prophylaxis to its workforce, with the goal of minimal disruption of patient care, here called hospital "self-prophylaxis." We created a new discrete-event simulation model of a hypothetical hospital wing to compare the operational charateristics of standard single-line, "first-come, first-served" dispensing clinics with those of 2 staff management strategies that can dramatically reduce staff waiting time while centralizing dispensing around existing pharmacy distribution points. PMID- 17464929 TI - Quality of hand hygiene in a pediatric hospital in Rio de Janeiro, Brazil. AB - We assessed the quality of hand hygiene among healthcare workers at a pediatrics hospital in Rio de Janeiro, Brazil. Hand hygiene was performed in 491 (34%) of 1,455 opportunities. Of these hand hygiene events, correct performance was observed in only 173 (35%). Multivariate analysis revealed that correct performance of hand hygiene was associated with the use of an alcohol-based product and a lack of jewelry (for all events) and employment in an infirmary with a comparatively higher ratio of nurses to patients (for events involving nurses). PMID- 17464930 TI - A neonatal specialist with recurrent methicillin-resistant Staphylococcus aureus (MRSA) carriage implicated in the transmission of MRSA to newborns. AB - This study reports an investigation of outbreaks of methicillin-resistant Staphylococcus aureus (MRSA) infection and colonization involving 17 newborns in the neonatal unit of a teaching hospital. A neonatal specialist colonized with MRSA that eventually became mupirocin-resistant was implicated as a recurrent source of transmission of MRSA to newborns. PMID- 17464931 TI - Incidence of parenterally transmitted acute viral hepatitis among healthcare workers in Italy. PMID- 17464932 TI - Conversion of prevalence survey data on nosocomial infections to incidence estimates: a simplified tool for surveillance? AB - We calculated the incidence of nosocomial infection in 2 intensive care units (ICUs) on the basis of prevalence data recorded from 1997 through 2002 and compared these estimates to cumulative incidences measured in the 2 ICUs during the same period to investigate the feasibility and the reliability of converting prevalence data to incidence estimates. Decreases in the calculated and measured incidences over time in the ICUs were found to be statistically significantly related. PMID- 17464933 TI - Pseudo-outbreak of Acinetobacter lwoffii infection in a tertiary care center in Thailand. AB - We describe a pseudo-outbreak of Acinetobacter lwoffii infection that was recognized early. The pseudo-outbreak involved 16 patients and occurred 3.5 months after the GNS-506 Vitek automated system was introduced in the microbiology laboratory. Prompt confirmation of incomplete use of the automated system's algorithm as the point source of the misidentified A. lwoffii clinical isolates averted a full outbreak investigation and excess use of infection control resources. PMID- 17464934 TI - Medication dosing errors in hospitalized patients with renal impairment: a study in Palestine. AB - BACKGROUND AND AIM: Reduced renal function requires dose adjustment for certain drugs to avoid toxicity. The aim of this study was to determine whether appropriate dosage adjustments were made for drugs that are nephrotoxic, excreted, or metabolized (TEM medications) by the kidney in patients with renal impairment. METHODOLOGY: A cross-sectional study of a group of hospitalized patients was carried out at Al-Watni governmental hospital, Nablus, Palestine. All patients with creatinine clearance A; 1155G>A; 1156+5_1156+8delGTAA], p.[G354R; L385L; missplicing], designated "complex E/I 10"). The G102S mutation was common in Type 1 GD patients and controls ( approximately 30% of alleles). In contrast, the E74K mutation was rare, present only in three Type 1 GD patients ( approximately 1% of alleles), all of Ashkenazi Jewish (AJ) descent, but it was not found in normal controls. The complex E/I-10 mutation occurred in two Caribbean Hispanic/African Type 1 GD patients and was present in 0 to 6% of alleles among normal controls from different populations. In vitro expression demonstrated that the E74K and G102S alleles had approximately 51% and approximately 23% of wild-type Chito catalytic efficiency, respectively. Expression of the G354R allele alone or with the L385L silent substitution did not produce detectable Chito activity or protein. RNA studies indicated that the complex E/I-10 allele also caused missplicing. Recognition of these mutations, particularly G102S, will facilitate the use and interpretation of plasma Chito activities for disease diagnosis, estimating disease severity, and monitoring therapeutic efficacy in GD. PMID- 17464954 TI - High-throughput enhancer trap by remobilization of transposon Minos in Ciona intestinalis. AB - The enhancer trap approach utilizing transposons yields us information about gene functions and gene expression patterns. In the ascidian Ciona intestinalis, transposon-based transgenesis and insertional mutagenesis were achieved with a Tc1/mariner transposon Minos. We report development of a novel technique for enhancer trap in C. intestinalis. This technique uses remobilization of Minos in the Ciona genome. A Minos vector for enhancer trap was constructed and a tandem array insertion of the vector was introduced into the Ciona genome to create a mutator line. Minos was remobilized in Ciona chromosomes to create new insertions by providing transposases. These transposase-introduced animals were crossed with wild-type animals. Nearly 80% of F1 families showed novel GFP expression patterns. This high-throughput enhancer trap screen will be useful to create new marker transgenic lines showing reporter gene expression in specific tissues and to identify novel patterns of gene expression. PMID- 17464955 TI - Molecular exchange through membranes of poly(2-vinylpyridine-block-ethylene oxide) vesicles. AB - The molecular exchange of tracer molecules through the membranes of dispersed vesicles of the block copolymer poly(2-vinylpyridine-block-ethylene oxide) was studied by using NMR spectroscopy combined with pulsed field gradients. The hydrodynamic radius of the tracer molecules was varied systematically to obtain a permeability profile of the vesicle membrane. In addition, the effect of system parameters, such as temperature, pH value, vesicle size, and thickness of the vesicle membrane, was studied. In the case of rapid exchange with average residence times significantly smaller than 10 s, the permeation is observed under equilibrium conditions and the data are analyzed by using a simple analytical approach. For slow exchange processes with average residence times above 10 s, the permeation is monitored in a time-resolved measurement under nonequilibrium conditions. Generally, the transmembrane exchange rate of the tracer clearly depends on its hydrodynamic radius. The characteristics of this dependence indicate the presence of two different mechanisms of membrane penetration, one dominating for smaller and one for larger tracer molecules, respectively. The exchange rate also shows a significant dependence on the bilayer thickness and on the vesicle diameter. By contrast, no variation of the membrane permeability with the temperature or the pH value could be detected as long as the vesicles remain stable. PMID- 17464956 TI - Self-repairing coatings containing active nanoreservoirs. AB - Nanocontainers with the ability to release encapsulated active materials in a controlled way can be employed to develop a new family of self-repairing multifunctional coatings, which possess not only passive functionality but also rapid feedback activity in response to changes in local environment. Several approaches to fabricate self-repairing coatings on plastic and metal substrates were surveyed. The release of the active materials occurs only when triggered, which prevents leakage of the active component out of the coating and increases coating durability. This Review also covers some principles and recent developments in the fabrication of nanocontainers with good compatibility with the coating components, the possibility to encapsulate and upkeep active material, and permeability properties of the shell controlled by external stimuli. Depending on the nature of the sensitive components introduced into the container shell, reversible and irreversible changes of the shell permeability can be induced by various stimuli. Different responses can be then observed varying from fine effects like tunable permeability to more drastic ones like total rupture of the container shell. PMID- 17464957 TI - Triazole: the keystone in glycosylated molecular architectures constructed by a click reaction. AB - The copper(I)-catalyzed modern version of the Huisgen-type azide-alkyne cycloaddition to give a 1,4-disubstituted 1,2,3-triazole unit is introduced as a powerful ligation method for glycoconjugation. Owing to its high chemoselectivity and tolerance of a variety of reaction conditions, this highly atom-economic and efficient coupling reaction is especially useful for the effective construction of complex glycosylated structures such as clusters, dendrimers, polymers, peptides, and macrocycles. In all cases the triazole ring plays a key role by locking into position the various parts of these molecular architectures. The examples reported and briefly discussed in this short review highlight the use of this reaction in carbohydrate chemistry and pave the way to further developments and applications. PMID- 17464958 TI - Synthesis of pyrazolo[4',3':5,6]pyrano[2,3-d]pyrimidine derivatives for antiviral evaluation. AB - 6-Amino-3-methyl-4-(4-nitrophenyl)-2,4-dihydropyrano[2,3-c]pyrazole-5 carbonitrile (1) was used as a precursor for preparation of some novel 3,7 dimethyl-4-(4-nitrophenyl)-2,4-dihydropyrazolo[4',3':5,6]pyrano[2,3-d]pyrimidine derivatives 3-6, and some of their corresponding N(2)- and C(5)-S-acyclic nucleosides 7 and 8. Furthermore, the preparation of 5-amino-1-[3,7-dimethyl-4-(4 nitrophenyl)-2,4-dihydropyrazolo[4',3':5,6]pyrano[2,3-d]pyrimidin-5-yl]-1H pyrazole derivatives 10-16 were described. Some of the prepared products were selected and tested for antiviral activity against Herpes Simplex Virus type-1 (HSV-1). PMID- 17464959 TI - Surfactant-coated single-walled carbon nanotubes as a novel pseudostationary phase in capillary EKC. AB - The analytical potential of the use of surfactant-coated single-walled carbon nanotubes (SC-SWNTs) as pseudostationary phase in CE is described. The pseudostationary phase shows an efficient alternative in enhancing electrochromatographic resolution of compounds which are capable of interacting with a nanotube surface, such as aromatic compounds. In general, the resolution is enhanced by increasing nanotube concentration in the buffer but the maximum amount of SWNTs that can be added to background electrolyte was found limited by compatibility with the UV/visible detection. As an alternative, a low-extension partial filling was used, consisting of the introduction into the capillary of concentrated SC-SWNT, just before the sample, with a plug length similar to the sample one. This has been showed as a reliable procedure in increasing resolution and sensitivity by sweeping phenomena. Finally, the potential of SC-SWNTs to perform chiral separations is discussed. PMID- 17464960 TI - Sample stacking for the analysis of catechins by microemulsion EKC. AB - In this study, an on-line concentration method, ASEI (anion-selective exhaustive injection)-sweeping technology which was coupled with microemulsion EKC (MEEKC), was used to analyze and detect six catechins ((-)-epicatechin, (+)-catechin, (-) epigallocatechin gallate, (-)-epicatechin gallate, (-)-epigallocatechin, and (-) gallocatechin). In addition to the effects of the buffer pH and electrolyte concentration on stacking, the compositions of microemulsion (types of oil phase, and types and levels of cosurfactant) also dominated the stacking effect of catechins. In MEEKC, the effect of the type of oil in microemulsion on separation mechanism is often unclear. This study had demonstrated that the oil type in microemulsion indeed altered the affinity of oil droplets with analytes. Finally, this proposed ASEI-sweeping MEEKC method was able to detect trace level of catechins in food products that was not previously possible by a normal MEEKC method. PMID- 17464961 TI - Optimization of CEC for simultaneous determination of eleven nucleosides and nucleobases in Cordyceps using central composite design. AB - A CEC method is described for the simultaneous determination of 11 nucleosides and nucleobases including cytosine, uracil, uridine, hypoxanthine, 2' deoxyuridine, inosine, guanosine, thymidine, adenine, adenosine, and cordycepin in Cordyceps using 5-chlorocytosine arabinoside as internal standard (IS). Chemometric optimization based on central composite design was employed to find the optimum conditions. The factors for optimization were defined as three parameters: voltage, pH, and concentration of ACN as organic modifier. The resolution (R(s)) between inosine and guanosine, as well as the entire run time were employed to evaluate the response function. A running buffer composed of 4 mM ammonium acetate and 2 mM triethylamine (TEA) adjusted to pH 5.3 using acetic acid, and containing 3% ACN as modifier, with gradient voltage (0-4 min: 20 kV, 4 12 min: linear gradient from 20 to 30 kV; 12-16 min: 30 kV) were found to be the optimum conditions for the separation. Separation of the 11 investigated compounds and 5-chlorocytosine arabinoside was achieved within 16 min. The contents of the 11 compounds in natural and cultured Cordyceps sinensis, and cultured Cordyceps militaris were also compared. The result showed that CEC is an efficient method for analysis of nucleosides and nucleobases in Cordyceps, which is helpful to control the quality of this valued traditional Chinese medicine. PMID- 17464962 TI - Two-chiral-component microemulsion electrokinetic chromatography-chiral surfactant and chiral oil: part 1. dibutyl tartrate. AB - The first simultaneous use of a chiral surfactant and a chiral oil for microemulsion EKC (MEEKC) is reported. Six stereochemical combinations of dodecoxycarbonylvaline (DDCV: R, S, or racemic, 2.00% w/v), racemic 2-hexanol (1.65% v/v), and dibutyl tartrate (D, L, or racemic, 1.23% v/v) were examined as chiral pseudostationary phases (PSPs) for the separation of six pairs of pharmaceutical enantiomers: pseudoephedrine, ephedrine, N-methyl ephedrine, metoprolol, synephrine, and atenolol. Subtle differences were observed for three chromatographic figures of merit (alpha(enant), alpha(meth), k) among the chiral microemulsions; a moderate difference was observed for efficiency (N) and elution range. Dual-chirality microemulsions provided both the largest and smallest enantioselectivities, due to small positive and negative synergies between the chiral microemulsion components. For the ephedrine family of compounds, dual chiral microemulsions with surfactant and oil in opposite stereochemical configurations provided higher enantioselectivities than the single-chiral component microemulsion (RXX), whereas dual-chiral microemulsions with surfactant and oil in the same stereochemical configurations provided lower enantioselectivities than RXX. Slight to moderate enantioselective synergies were confirmed using a thermodynamic model. Efficiencies observed with microemulsions comprised of racemic dibutyl tartrate or dibutyl-D-tartrate were significantly higher than those obtained with dibutyl-L-tartrate, with an average difference in plate count of about 25 000. Finally, one two-chiral-component microemulsion (RXS) provided significantly better resolution than the remaining one- and two chiral-component microemulsions for the ephedrine-based compounds, but only slightly better or equivalent resolution for non-ephedrine compounds. PMID- 17464963 TI - Electrokinetic partial filling technique as a powerful tool for enantiomeric separation of DL-lactic acid by CE with contactless conductivity detection. AB - A modified partial filling method for chiral separation of DL-lactic acid as the model chiral compound with vancomycin chloride as the chiral selector was developed by CE with contactless conductivity detection. Electrokinetic partial filling technique (EK-PFT) was used as an alternative method to the conventional hydrodynamic partial filling method. EK-PFT, in contrast to the hydrodynamic partial filling technique, allowed the removal of the chloride counterions from the chiral selector which otherwise led to poor sensitivity in conductivity detection. The baseline separation of DL-lactic acid as the model analyte was achieved in 5 min in a polyacrylamide-coated capillary. The best resolution was achieved by electrokinetic partial filling of vancomycin cations from the injection solution containing 5 mmol/L oxalate L-histidinium at pH 4.5 with 10 mmol/L vancomycin chloride. Computer simulation was used to explain the observed phenomena in the boundary between the inject vial and the capillary during the EK PFT of vancomycin cations. PMID- 17464964 TI - Synthesis and anti-HIV-1 activity of new MKC-442 analogues with an alkynyl substituted 6-benzyl group. AB - Synthesis and antiviral activities are reported of a series of 6-(3-alkynyl benzyl)-substituted analogues of MKC-442 (6-benzyl-1-(ethoxymethyl)-5 isopropyluracil), a highly potent agent against HIV. The 3-alkynyl group is assumed to give a better stacking of the substituted benzyl group to reverse transcriptase (RT) and this was believed to improve antiviral activity against HIV-1. The bromo derivatives, 5-alkyl-6-(3-bromo-benzyl)-1-ethoxymethyl derivatives 7a, b and 5-alkyl-6-(3-bromobenzyl)-1-allyloxymethyl derivatives 9a, b, showed activity against HIV on the same level as their corresponding analogues 10a-d with a 3-trimethylsilylalkynylbenzyl substituent and their desilylated analogues 11a-d. However, they all showed activity against HIV-1 wild type in the range of more than 10fold lower than the one of MKC-442. Moderate activity against Y181C and Y181C + K103N mutated strains was also observed and, in some cases, they were marginally better than those found for MKC-442. A few amino-DABO and S-DABO analogues were also synthesized but they were found to be inactive against HIV. PMID- 17464965 TI - Synthesis, structure and hypoxic cytotoxicity of 3-amino-1,2,4-benzotriazine-1,4 dioxide derivatives. AB - A series of novel 3-amino-1,2,4-benzotriazine-1,4-dioxide derivatives were synthesized and screened for their in vitro cytotoxicity against promyelocytic leukemia HL-60, androgen-independent prostate tumor PC3, hepatocellular carcinoma Bel-7402, human esophagus tumor ECA-109, and human breast tumor MCF-7 cell lines in hypoxia and in normoxia. Most compounds showed higher cytotoxic activity both in hypoxia and in normoxia. Among them, compounds 61 and 62 showed more potent cytotoxic activity and hypoxic selectivity when compared to tirapazamine. PMID- 17464966 TI - Anti-mitotic activity of colchicine and the structural basis for its interaction with tubulin. AB - In this review, an attempt has been made to throw light on the mechanism of action of colchicine and its different analogs as anti-cancer agents. Colchicine interacts with tubulin and perturbs the assembly dynamics of microtubules. Though its use has been limited because of its toxicity, colchicine can still be used as a lead compound for the generation of potent anti-cancer drugs. Colchicine binds to tubulin in a poorly reversible manner with high activation energy. The binding interaction is favored entropically. In contrast, binding of its simple analogs AC or DAAC is enthalpically favored and commences with comparatively low activation energy. Colchicine-tubulin interaction, which is normally pH dependent, has been found to be independent of pH in the presence of microtubule associated proteins, salts or upon cleavage of carboxy termini of tubulin. Biphasic kinetics of colchicines-tubulin interaction has been explained in light of the variation in the residues around the drug-binding site on beta-tubulin. Using the crystal structure of the tubulin-DAMAcolchicine complex, a detailed discussion on the pharmacophore concept that explains the variation of affinity for different colchicine site inhibitors (CSI) has been discussed. PMID- 17464967 TI - Biological agents for ulcerative colitis: hypes and hopes. AB - Ulcerative colitis (UC) is an idiopathic chronic inflammatory disease of the colonic mucosa. Over the last decade, the increasing knowledge on the pathogenic mechanisms underlying intestinal inflammation has led to the development of a number of biological agents, mainly addressed to molecules and/or pathways demonstrated to have a pathogenic role in UC. In UC, clinical course and therapeutic decisions mainly depend on disease activity and extent. While therapeutic approach to mild-to-moderate UC by using aminosalicylates and corticosteroids has been well established, treatment of severe UC is far from being satisfactory. A severe attack of UC remains a challenge to be managed jointly by gastroenterology, surgery, and intensive care units. However, the recent introduction of biological therapies has led to promising changes in the management of UC patients. Aim of this paper is to review the recent advances and future perspectives for the use of biological agents in UC. PMID- 17464968 TI - Validation of critical points in the electron density as descriptors by building quantitative structure-property relationships for the atomic polar tensor. AB - A crucial component of research in the field of quantitative structure activity/property relationships is the identification of molecular descriptors relevant to the activity or property of interest. Descriptors based on the topology of the electron density as formulated in Bader's theory of atoms in molecules are investigated in detail in this work. In a model study, the authors investigate their ability to predict the atomic polar tensor (the gradient of the molecular dipole moment), which contains information on the vibrational intensities in infrared spectroscopy and constitutes a scheme for partitioning the total charge distribution into atomic charges. The atomic polar tensor may therefore be used to investigate whether the descriptors give adequate information on the local electronic structure in the molecule. Both the trace of the atomic polar tensor and for planar molecules its out-of-plane component may be interpreted as definitions of atomic charges suitable for prediction. Hydrogen and carbon atoms in a set of 60 aromatic compounds with various substituents have been studied. Excellent results for prediction of hydrogen and carbon charges have been achieved with cross-validated squared correlation coefficients between predicted and theoretical values varying from 0.92 and 0.977 for the most complex set of substituents when the value, Laplacian, and ellipticity of the electron density in the bond critical points are used as descriptors. The carbon charges defined from the trace of the atomic polar tensor are correlated with its out-of plane component whereas such relationship is not observed for the hydrogen charges studied in this work. PMID- 17464969 TI - Diagnostic accuracy of the Multistix 8 SG reagent strip in diagnosis of spontaneous bacterial peritonitis. AB - Recent studies have shown that the diagnosis of spontaneous bacterial peritonitis (SBP) can be rapidly obtained using leukocyte esterase reagent strips. However, published studies were restricted to one or two centers, and the number of patients with SBP was thus limited. The aims of the current prospective multicenter study were: (1) to assess the diagnostic accuracy of the Multistix 8SG urine test for the diagnosis of SBP; and (2) to assess the prevalence of SBP. From January to May 2004, 2 reactive strips were tested independently in inpatients with cirrhosis and in outpatients undergoing paracentesis. Cultures of ascitic fluid were performed at the bedside using aerobic and anaerobic blood culture bottles. Two thousand one hundred twenty-three paracenteses were performed in 1,041 patients from 70 centers. One hundred seventeen samples, obtained from 91 patients, had ascites polymorphonuclear cell (PMN) counts>or=250/microl (range, 250-34,000), among which 56 were associated with positive ascitic fluid cultures. The prevalence of SBP was 5.5% in the whole population, 9% in inpatients, and 1.3% in outpatients (P<0.0001). The prevalence of SBP was 0.57% in asymptomatic outpatients versus 2.4% in symptomatic outpatients (P=0.04). Using a threshold of 2+ for positivity of the reagent strip, sensitivity was 45.3% for the diagnosis of SBP, specificity was 99.2%, positive predictive value was 77.9%, and negative predictive value was 96.9%. CONCLUSION: This study confirms the low prevalence of SBP in asymptomatic outpatients according to a priori defined criteria, and indicates an absence of diagnostic efficacy for this specific strip test. PMID- 17464970 TI - Racial disparities in the management of hospitalized patients with cirrhosis and complications of portal hypertension: a national study. AB - Having complications of portal hypertension is a harbinger of decompensated cirrhosis and warrants consideration for liver transplantation (LT). Racial disparities in LT have been reported. We sought to characterize disparities in the performing of surgical and endoscopic procedures among hospitalized patients with complications of portal hypertension. We queried the Nationwide Inpatient Sample from 1998 to 2003 to identify patients with cirrhosis and complications of portal hypertension. Logistic regression controlling for confounders was used to evaluate race as a predictor of undergoing a portosystemic shunt and LT and of dying in the hospital. Compared to whites, the adjusted odds ratios of receiving a portosystemic shunt were 0.37 (95% CI: 0.27-0.51) and 0.69 (95% CI: 0.54-0.88) for African Americans (AAs) and Hispanics, respectively. AAs with variceal bleeding were more likely to have endoscopic variceal hemostasis delayed more than 24 hours after admission than were whites (OR 1.6; 95% CI: 1.2-2.1). The adjusted odds ratios of undergoing LT were 0.32 (95% CI:0.20-0.52) and 0.46 (95% CI: 0.25-0.83) for AAs and Hispanics, respectively. Compared to whites, AAs experienced higher in-hospital mortality (OR 1.12; 95% CI: 1.01-1.24), whereas Hispanics had a lower risk of death (OR 0.83; 95% CI: 0.75-0.92). Among variceal bleeders, the odds ratio of death for AAs was 1.7 (95% CI: 1.2-2.4) compared to whites. CONCLUSION: AAs and Hispanics hospitalized for complications of portal hypertension were less likely to undergo a palliative shunt or LT than whites, which may contribute to the higher in-hospital mortality of AAs. Further studies are warranted to elucidate the mechanisms of these exploratory findings. PMID- 17464971 TI - Liver stiffness measurement predicts severe portal hypertension in patients with HCV-related cirrhosis. AB - Measurement of hepatic venous pressure gradient (HVPG) is a standard method for the assessment of portal pressure and correlates with the occurrence of its complications. Liver stiffness measurement (LSM) has been proposed as a noninvasive technique for the prediction of the complications of cirrhosis. In this study, we evaluated the ability of LSM to predict severe portal hypertension compared with that of HVPG in 61 consecutive patients with HCV-related chronic liver disease. A strong relationship between LSM and HVPG measurements was found in the overall population (r=0.81, P<0.0001). However, although the correlation was excellent for HVPG values less than 10 or 12 mm Hg (r=0.81, P=0.0003 and r=0.91, P<0.0001, respectively), linear regression analysis was not optimal for HVPG values>or=10 mm Hg (r2=0.35, P<0.0001) or>or=12 mm Hg (r2=0.17, P=0.02). The AUROC for the prediction of HVPG>or=10 and >or=12 mm Hg were 0.99 and 0.92, respectively and at LSM cutoff values of 13.6 kPa and 17.6 kPa, sensitivity was 97% and 94%, respectively. In patients with cirrhosis, LSM positively correlated with the presence of esophageal varices (P=0.002), although no correlation between LSM and esophageal varices size was detected. The area under the ROC for the prediction of EV was 0.76 and at a LSM cutoff value of 17.6 kPa sensitivity was 90%. CONCLUSION: LSM represents a non-invasive tool for the identification of chronic liver disease patients with clinically significant or severe portal hypertension and could be employed for screening patients to be subjected to standard investigations including upper GI endoscopy and hemodynamic studies. PMID- 17464972 TI - WNT/beta-catenin signaling in liver health and disease. AB - Wnt/beta-catenin signaling is emerging as a forerunner for its critical roles in many facets of human biology. Its roles in embryogenesis, organogenesis, and maintaining tissue and organ homeostasis demonstrate its munificent character. Its roles in pathological conditions such as cancer and other human disorders such as inflammatory disorders and fibrosis reveal its villainous disposition. In liver, it also maintains its dual personality and is clearly of essence in several physiological events such as development, regeneration, and growth. Its aberrant activation is also evident in many different tumors of the liver, and recent studies are beginning to identify its role in additional hepatic pathological conditions. It is contributing to liver physiology and pathology by regulating various basic cellular events, including differentiation, proliferation, survival, oxidative stress, morphogenesis, and others. This review discusses the contribution of the Wnt/beta-catenin signaling pathway in these events and simultaneously provides an essential overview of the major developments in the field of Wnt/beta-catenin and liver pathobiology. In addition, areas that are currently deficient or understudied are identified and discussed along with the avenues of translational and clinical relevance. PMID- 17464973 TI - Role of S-adenosyl-L-methionine in liver health and injury. AB - S-adenosylmethionine (SAMe) has rapidly moved from being a methyl donor to a key metabolite that regulates hepatocyte growth, death, and differentiation. Biosynthesis of SAMe occurs in all mammalian cells as the first step in methionine catabolism in a reaction catalyzed by methionine adenosyltransferase (MAT). Decreased hepatic SAMe biosynthesis is a consequence of all forms of chronic liver injury. In an animal model of chronic liver SAMe deficiency, the liver is predisposed to further injury and develops spontaneous steatohepatitis and hepatocellular carcinoma. However, impaired SAMe metabolism, which occurs in patients with mutations of glycine N-methyltransferase (GNMT), can also lead to liver injury. This suggest that hepatic SAMe level needs to be maintained within a certain range, and deficiency or excess can both lead to abnormality. SAMe treatment in experimental animal models of liver injury shows hepatoprotective properties. Meta-analyses also show it is effective in patients with cholestatic liver diseases. Recent data show that exogenous SAMe can regulate hepatocyte growth and death, independent of its role as a methyl donor. This raises the question of its mechanism of action when used pharmacologically. Indeed, many of its actions can be recapitulated by methylthioadenosine (MTA), a by-product of SAMe that is not a methyl donor. A better understanding of why liver injury occurs when SAMe homeostasis is perturbed and mechanisms of action of pharmacologic doses of SAMe are essential in defining which patients will benefit from its use. PMID- 17464974 TI - Molecular pathogenesis of alpha-1-antitrypsin deficiency-associated liver disease: a meeting review. AB - In recent years, we have witnessed several important paradigm shifts in understanding the molecular basis of liver disease in alpha-1-antitrypsin (AT) deficiency. These shifts have become possible as a result of a number of advances in research on the cell biology of aggregation-prone mutant proteins and in research on the pathobiological mechanisms of liver disease in general. Late breaking research in these areas was the subject of an AASLD/Alpha-1 Foundation Single Topic Conference in Atlanta, Georgia, on January 26 to 28, 2006. The conference was titled "Alpha-1-Antitrypsin Deficiency and Other Liver Diseases Caused by Aggregated Proteins." Investigators from all over the world, representing a broad array of scientific disciplines and perspectives, discussed the pathobiology of AT deficiency, mechanisms of cell injury in diseases associated with aggregation-prone proteins, pathways by which cells respond to protein aggregation and mislocalization, and mechanisms of liver injury in general and in diseases related to AT deficiency. A session of the meeting was devoted to novel therapeutic strategies being developed for AT deficiency as well as to strategies either in development or already being applied to the class of diseases associated with mutant proteins. PMID- 17464975 TI - Bone morphogenetic protein-7 in focus: a member of the transforming growth factor beta superfamily is implicated in the maintenance of liver health. PMID- 17464976 TI - Today's hepatic stellate cells: not your father's sternzellen. PMID- 17464977 TI - Liver anti-inflammatory pathways: potential ways to control liver damage. PMID- 17464978 TI - Low incidence of HCV reinfection: exposure, testing frequency, or protective immunity? PMID- 17464980 TI - Hepatitis D virus infection--not a vanishing disease in Europe! PMID- 17464982 TI - Impact of disease severity on outcome of antiviral therapy in treatment-naive patients with chronic hepatitis C. PMID- 17464984 TI - Transient elastography for diagnosis of portal hypertension in liver cirrhosis: is there still a role for hepatic venous pressure gradient measurement? PMID- 17464985 TI - Bile ductules and stromal cells express hedgehog ligands and/or hedgehog target genes in primary biliary cirrhosis. AB - Indian Hedgehog (Ihh) regulates tissue morphogenesis. Hedgehog (Hh) activity has been demonstrated in human cholangiocarcinoma and hepatocellular carcinoma lines, and in myofibroblasts and progenitors from adult rodent livers. We evaluated Hh pathway involvement in the response to biliary injury in primary biliary cirrhosis (PBC). Liver sections from 3 PBC patients and 3 controls without liver disease were studied. Immunohistochemistry was used to determine if cells that accumulate in PBC livers express Ihh or Hh-target genes including the Hh receptor, Patched (Ptc), and the Hh-transcriptional activator glioblastoma (Gli) 2. Positive cells were further identified by costaining for cytokeratin (CK) 19, a biliary marker, or OV6, a hepatic progenitor marker. In all subjects, Gli2 and Ptc expression localized in portal areas. The numbers of Gli2- or Ptc-expressing cells/portal triad were each 10-fold greater in patients with PBC than in controls (P<0.05). In PBC livers, some CK19+ cells coexpressed Gli2 or Ptc. Many stromal fibroblastic cells were also Gli2+. Strong Ihh expression was detected in most bile ductular cells. Scattered stromal cells also expressed Ihh. The number of Ihh+ cells/portal triad was 6-fold greater in PBC livers than controls (P<0.05). OV6+ progenitors increased significantly in PBC livers, and some of these cells coexpressed Ihh, Ptc, and/or Gli2. CONCLUSION: This is the first direct evidence that noncancerous, adult human livers harbor several types of cells that produce and/or respond to Hh ligands. Such Hh-responsive cells accumulate during the fibroproliferative response to chronic cholestatic liver injury, suggesting a role for Hh signaling in this process. PMID- 17464986 TI - Polymorphism in microsomal triglyceride transfer protein: a link between liver disease and atherogenic postprandial lipid profile in NASH? AB - Nonalcoholic fatty liver disease (NAFLD) is emerging as an independent cardiovascular risk factor, but mechanism(s) linking fatty liver to atherosclerosis are unknown. Microsomal triglyceride transfer protein (MTP) -493 G/T polymorphism modulates circulating lipid and lipoprotein levels in different subsets and has been linked to NAFLD. The impact of MTP -493 G/T polymorphism, adipokines, and diet on postprandial lipoprotein profile and liver disease was assessed in nonalcoholic steatohepatitis (NASH). Plasma lipids, triglyceride-rich lipoprotein subfractions, high-density lipoprotein-C (HDL-C), and oxidized low density lipoprotein (LDL) after an oral fat load were cross-sectionally correlated to MTP -493 G/T polymorphism, dietary habits, adipokines, and liver histology in 29 nonobese nondiabetic patients with NASH and 27 healthy controls. The severity of liver histology, the magnitude of triglycerides (Tg), free fatty acid (FFA), and LDL-conjugated diene responses, and the fall in HDL-C and apoA1 were significantly higher in NASH G/G (66% of patients) than in the other genotypes, despite similar adipokine profile and degree of insulin resistance. Postprandial large intestinal very-low-density lipoprotein (VLDL) subfraction A increases independently predicted Tg (beta=0.48; P=.008), FFA (beta=0.47; P=0.010), HDL-C (beta=0.42; P=0.009), and LDL-conjugated diene (beta=0.52; P=0.002) responses. VLDL A apoB48 response was independently associated with liver steatosis (OR: 2.4; CI 1.7-9.6; P=0.031). Postprandial LDL-conjugated diene response predicted severe necroinflammation (OR: 3.3; CI 1.4-9.7; P=0.016) and fibrosis (OR: 2.8; CI 1.0-8,5; P=0.030); postprandial apoA1 fall predicts severe fibrosis (OR: 2.1; CI: 1.5-6.1; P=0.015). CONCLUSION: MTP -493 G/T polymorphism may impact NASH by modulating postprandial lipemia and lipoprotein metabolism; homozygous GG carriers have a more atherogenic postprandial lipid profile than the other genotypes, independently of adipokines and insulin resistance. PMID- 17464987 TI - CCAAT/enhancing binding protein beta deletion in mice attenuates inflammation, endoplasmic reticulum stress, and lipid accumulation in diet-induced nonalcoholic steatohepatitis. AB - Nonalcoholic steatohepatitis (NASH) is characterized by steatosis, inflammation, and oxidative stress. To investigate whether the transcription factor CCAAT/Enhancer binding protein (C/EBPbeta) is involved in the development of NASH, C57BL/6J wild-type (WT) or C/EBPbeta knockout (C/EBPbeta-/-) mice were fed either a methionine and choline deficient (MCD) diet or standard chow. These WT mice fed a MCD diet for 4 weeks showed a 2- to 3-fold increase in liver C/EBPbeta messenger RNA and protein, along with increased expression of lipogenic genes peroxisome proliferators-activated receptor gamma and Fas. WT mice also showed increased levels of the endoplasmic reticulum stress pathway proteins phosphorylated eukaryotic translation initiation factor alpha, phosphorylated pancreatic endoplasmic reticulum kinase, and C/EBP homologous protein, along with inflammatory markers phosphorylated nuclear factor kappaB and phosphorylated C jun N-terminal kinase compared to chow-fed controls. Cytochrome P450 2E1 protein and acetyl coA oxidase messenger RNA involved in hepatic lipid peroxidation were also markedly increased in WT MCD diet-fed group. In contrast, C/EBPbeta-/- mice fed a MCD diet showed a 60% reduction in hepatic triglyceride accumulation and decreased liver injury as evidenced by reduced serum alanine aminotransferase and aspartate aminotransferase levels, and by H&E staining. Immunoblots and real-time qPCR data revealed a significant reduction in expression of stress related proteins and lipogenic genes in MCD diet-fed C/EBPbeta-/- mice. Furthermore, circulating TNFalpha and expression of acute phase response proteins CRP and SAP were significantly lower in C/EBPbeta-/- mice compared to WT mice. Conversely, C/EBPbeta over-expression in livers of WT mice increased steatosis, nuclear factor-kappaB, and endoplasmic reticulum stress, similar to MCD diet-fed mice. CONCLUSION: Taken together, these data suggest a previously unappreciated molecular link between C/EBPbeta, hepatic steatosis and inflammation and suggest that increased C/EBPbeta expression may be an important factor underlying events leading to NASH. PMID- 17464988 TI - Hepatocyte-specific Gclc deletion leads to rapid onset of steatosis with mitochondrial injury and liver failure. AB - Oxidative stress is considered to be a critical mediator in liver injury of various etiologies. Depletion of glutathione (GSH), the major antioxidant in liver, has been associated with numerous liver diseases. To explore the specific role of hepatic GSH in vivo, we targeted Gclc, a gene essential for GSH synthesis, so that it was flanked by loxP sites and used the albumin-cyclization recombination (Alb-Cre) transgene to disrupt the Gclc gene specifically in hepatocytes. Deletion within the Gclc gene neared completion by postnatal day (PND)14, and loss of GCLC protein was complete by PND21. Cellular GSH was progressively depleted between PND14 and PND28-although loss of mitochondrial GSH was less severe. Nevertheless, ultrastructural examination of liver revealed dramatic changes in mitochondrial morphology; these alterations were accompanied by striking decreases in mitochondrial function in vitro, cellular ATP, and a marked increase in lipid peroxidation. Plasma liver biochemistry tests from these mice were consistent with progressive severe parenchymal damage. Starting at PND21, livers from hepatocyte-specific Gclc knockout [Gclc(h/h)] mice showed histological features of hepatic steatosis; this included inflammation and hepatocyte death, which progressed in severity such that mice died at approximately 1 month of age due to complications from liver failure. CONCLUSION: GSH is essential for hepatic function and loss of hepatocyte GSH synthesis leads to steatosis with mitochondrial injury and hepatic failure. PMID- 17464989 TI - Tissue factor pathway inhibitor-2 as a frequently silenced tumor suppressor gene in hepatocellular carcinoma. AB - In HCC, inactivation of tumor suppressor genes plays a significant role in carcinogenesis. Apart from deletions and mutations, growing evidence has indicated that epigenetic alterations including aberrant promoter methylation and histone deacetylation are also implicated in inactivation of tumor suppressor genes. The goal of this study was to identify epigenetically silenced candidate tumor suppressor genes in human HCC by comparing the changes in oligonucleotide microarray gene expression profiles in HCC cell lines upon pharmacological treatment with the demethylating agent 5-Aza-2'-deoxycytidine (5-Aza-dC). By analyzing the gene expression profiles, we selected tissue factor pathway inhibitor-2 (TFPI-2), a Kunitz-type serine protease inhibitor, for validation and further characterization. Our results showed that TFPI-2 was frequently silenced in human HCC and HCC cell lines. TFPI-2 was significantly underexpressed in approximately 90% of primary HCCs when compared with their corresponding nontumorous livers. TFPI-2 promoter methylation was detected in 80% of HCC cell lines and 47% of human HCCs and was accompanied by reduced TFPI-2 messenger RNA expression. In addition, TFPI-2 expression in HCC cell lines can be robustly restored by combined treatment with 5-Aza-dC and histone deacetylase inhibitor trichostatin A. These findings indicate that TFPI-2 is frequently silenced in human HCC via epigenetic alterations, including promoter methylation and histone deacetylation. Moreover, ectopic overexpression of TFPI-2 significantly suppressed the proliferation and invasiveness of HCC cells. CONCLUSION: Our findings suggest that TFPI-2 is a candidate tumor suppressor gene in human HCC. PMID- 17464990 TI - Contrast-enhanced ultrasound of histologically proven liver hemangiomas. AB - Differentiation of small and atypical hemangiomas from other hepatic masses using imaging methods can be difficult, especially in patients with underlying malignant disease. Therefore, contrast-enhanced ultrasound was assessed in patients with histologically confirmed hemangiomas with respect to contrast enhancing kinetics and tumor characteristics. In 58 patients with indeterminate hepatic lesions demonstrated with at least 2 imaging methods (ultrasound/computed tomography/magnetic resonance imaging), ultrasound-guided liver biopsy revealed hemangioma. In all patients a hepatic neoplasm had been suspected because of underlying malignant disease (n=41), liver cirrhosis (n=15), or growth of the lesion (n=2). All patients underwent nonlinear, low mechanical index real-time contrast-enhanced ultrasound scanning with bolus injections of SonoVue. Peripheral nodular arterial enhancement was detected in 43 patients (74%), whereas the typical metastatic peripheral rim-like enhancement was not observed at all. Strong homogenous arterial enhancement was found in 9 of 58 (16%) patients. In 6 patients (10%), the arterial contrast enhancement pattern could not be determined because of the very small size of the lesions or fibrotic nodules. Forty-five (78%) of the hemangiomas showed homogenous centripetal filling within 180 seconds. CONCLUSION: Contrast-enhanced ultrasound demonstrates typical hemangioma imaging characteristics, that is, peripheral nodular contrast enhancement and iris-diaphragm sign in a high percentage of patients with undetermined lesions. This technique may therefore improve noninvasive functional characterization and differentiation of hemangiomas. PMID- 17464991 TI - Differential regulation of constitutive androstane receptor expression by hepatocyte nuclear factor4alpha isoforms. AB - Constitutive androstane receptor (CAR; NR1I3) controls the metabolism and elimination of endogenous and exogenous toxic compounds by up-regulating a battery of genes. In this work, we analyzed the expression of human CAR (hCAR) in normal liver during development and in hepatocellular carcinoma (HCC) and investigated the effect of hepatocyte nuclear factor 4alpha isoforms (HNF4alpha1 and HNF4alpha7) on the hCAR gene promoter. By performing functional analysis of hCAR 5'-deletions including mutants, chromatin immunoprecipitation in human hepatocytes, electromobility shift and cotransfection assays, we identified a functional and species-conserved HNF4alpha response element (DR1: ccAGGCCTtTGCCCTga) at nucleotide -144. Both HNF4alpha isoforms bind to this element with similar affinity. However, HNF4alpha1 strongly enhanced hCAR promoter activity whereas HNF4alpha7 was a poor activator and acted as a repressor of HNF4alpha1-mediated transactivation of the hCAR promoter. PGC1alpha stimulated both HNF4alpha1-mediated and HNF4alpha7-mediated hCAR transactivation to the same extent, whereas SRC1 exhibited a marked specificity for HNF4alpha1. Transduction of human hepatocytes by HNF4alpha7-expressing lentivirus confirmed this finding. In addition, we observed a positive correlation between CAR and HNF4alpha1 mRNA levels in human liver samples during development, and an inverse correlation between CAR and HNF4alpha7 mRNA levels in HCC. These observations suggest that HNF4alpha1 positively regulates hCAR expression in normal developing and adult livers, whereas HNF4alpha7 represses hCAR gene expression in HCC. PMID- 17464992 TI - Twenty-four-week clevudine therapy showed potent and sustained antiviral activity in HBeAg-positive chronic hepatitis B. AB - Clevudine is a pyrimidine analogue with potent and sustained antiviral activity against HBV. The present study evaluated the safety and efficacy of 30 mg clevudine once daily for 24 weeks and assessed the durable antiviral response for 24 weeks after cessation of dosing. A total of 243 hepatitis B e antigen (HBeAg) positive chronic hepatitis B patients were randomized (3:1) to receive clevudine 30 mg once daily (n=182) or placebo (n=61) for 24 weeks. Patients were followed for a further 24 weeks off therapy. Median serum HBV DNA reductions from baseline at week 24 were 5.10 and 0.27 log10 copies/mL in the clevudine and placebo groups, respectively (P<0.0001). Viral suppression in the clevudine group was sustained off therapy, with 3.73 log10 reduction at week 34 and 2.02 log10 reduction at week 48. At week 24, 59.0% of patients in the clevudine group had undetectable serum HBV DNA levels by Amplicor PCR assay (less than 300 copies/mL). The proportion of patients who achieved normalization of alanine aminotransferase (ALT) levels was 68.2% in the clevudine group and 17.5% in the placebo group at week 24 (P<0.0001). ALT normalization in the clevudine group was well maintained during post-treatment follow-up period. The incidence of adverse events (AEs) was similar between the clevudine group and the placebo group. No resistance to clevudine was detected with 24 weeks of administration of drug. CONCLUSION: A 24-week clevudine therapy was well tolerated and showed potent and sustained antiviral effect without evidence of viral resistance during treatment period in HBeAg-positive chronic hepatitis B. PMID- 17464993 TI - Hepatitis B viral factors in HBeAg-negative carriers with persistently normal serum alanine aminotransferase levels. AB - Chronic hepatitis B patients with high-normal serum ALT (levels of 0.5-1x upper limit of normal) are still at risk of liver disease progression. We thus investigated the correlation between serum ALT level and hepatitis B viral factors in HBeAg-negative carriers with persistently normal serum ALT level (PNALT). Baseline clinical and virological features of 414 HBeAg-negative carriers, including 176 (42.5%) with low-normal ALT (levels of less than 0.5x upper limit of normal) and 238 (57.5%) with high-normal ALT, were compared. Compared with HBV carriers with low-normal ALT, those with high-normal ALT were older (41 vs. 37 years, P<0.001) and had a greater frequency of serum HBV DNA level>10(4) copies/ml (63.4% vs. 47.5%, P<0.001) as well as a higher prevalence of basal core promoter T1762/A1764 mutant (36.5% vs. 24.2%, P=0.01). Multivariate analysis showed that factors associated with a high-normal serum ALT level included male sex [odds ratio (OR), 1.82; 95% confidence interval (CI), 1.10 3.01, P=0.019], increasing age (OR, <30 years: 1, reference; 30-39 years: 2.43, 95% CI, 1.18-5.03, P=0.016; 40-49 years: 4.22, 95% CI, 1.99-8.93, P<0.001; >or=50 years: 4.06, 95% CI, 1.69-9.78, P=0.002) and serum HBV DNA level>10(4) copies/ml (OR, 1.83; 95% CI, 1.07-3.13, P=0.027). CONCLUSION: HBeAg-negative patients with persistently normal ALT are not a homogenous group, and those with high-normal ALT share some of the characteristics that have been associated with adverse long term outcomes. PMID- 17464994 TI - Met signals hepatocyte survival by preventing Fas-triggered FLIP degradation in a PI3k-Akt-dependent manner. AB - The FasL-Fas couple is a general death mediator whose activated signals lead to caspase-8 activation and apoptosis in adult hepatocytes. Suppression of caspase-8 activation and cell death is a protective mechanism modulated by the FLICE-Like Inhibitory Protein (FLIP). Although hepatocyte growth factor (HGF) and its receptor Met are known to mediate cell survival in developing livers, the molecular mechanisms involved in this process are poorly understood. We show here that Met activation by HGF impairs Fas-triggered apoptosis of primary embryonic hepatocytes and cell survival correlates with inhibition of caspase-8 and caspase 3 activities. Furthermore, we found that HGF treatment prevents degradation of FLIPL triggered by Fas activation. In contrast to this, Met activation does not modulate FLIPL levels and its stability in untreated cells, thus showing the specificity of this regulatory mechanism for embryonic hepatocyte survival. Knocking down FLIP expression abolishes the ability of Met to inhibit Fas triggered hepatocyte death, demonstrating the functional requirement of FLIP in HGF anti-apoptotic signals. By combining genetic and pharmacological approaches, we also demonstrate that the PI3K-Akt pathway is required in embryonic hepatocytes to prevent Fas-triggered FLIP degradation and death. Thus, Met acting on PI3K and Akt ensures high levels of FLIPL, and disruption of this pathway contributes to hepatic apoptosis and possibly to Fas-related liver diseases. PMID- 17464995 TI - Hepatocyte-conditioned medium sustains endothelial differentiation of human hematopoietic-endothelial progenitors. AB - Liver neo-angiogenesis plays a fundamental role in physiological and pathological processes such as regeneration, cirrhosis, autoimmune hepatitis, and alcoholic liver disease. How liver parenchymal cells influence angiogenesis is largely unknown. We studied the influence of soluble factors released by hepatocytes on hematopoietic and endothelial cell differentiation. Human CD34+ cells cultured for several weeks in a hepatocyte-conditioned medium gradually decrease the expression of CD34 and CD133 markers (i.e. after 4 weeks from 85% and 69%, respectively, to 6% and 3%, respectively), whereas expression of CD144 and CD14 cell markers increased (from 2% and 8%, respectively, to 54% and 55%, respectively). The cells' capacity to form hematopoietic colonies in methylcellulose declined with time, whereas they acquired endothelial morphology, expressed endothelial markers, and incorporated into newly forming vascular structures both in vitro and in vivo. Cultured single CD34+ cells formed colonies expressing both hematopoietic (CD45+) and endothelial (CD144+) markers, suggesting they constitute a bona fide hemangioblast population. CONCLUSION: This system allowed subsequent stages of differentiation of hematopoietic cells to endothelial cells to be defined, underlining the strict interrelationship between endothelial and hematopoietic cells in a hepatocyte environment. PMID- 17464996 TI - Directed differentiation of human embryonic stem cells into functional hepatic cells. AB - The differentiation capacity of human embryonic stem cells (hESCs) holds great promise for therapeutic applications. We report a novel three-stage method to efficiently direct the differentiation of human embryonic stem cells into hepatic cells in serum-free medium. Human ESCs were first differentiated into definitive endoderm cells by 3 days of Activin A treatment. Next, the presence of fibroblast growth factor-4 and bone morphogenetic protein-2 in the culture medium for 5 days induced efficient hepatic differentiation from definitive endoderm cells. Approximately 70% of the cells expressed the hepatic marker albumin. After 10 days of further in vitro maturation, these cells expressed the adult liver cell markers tyrosine aminotransferase, tryptophan oxygenase 2, phosphoenolpyruvate carboxykinase (PEPCK), Cyp7A1, Cyp3A4 and Cyp2B6. Furthermore, these cells exhibited functions associated with mature hepatocytes including albumin secretion, glycogen storage, indocyanine green, and low-density lipoprotein uptake, and inducible cytochrome P450 activity. When transplanted into CCl4 injured severe combined immunodeficiency mice, these cells integrated into the mouse liver and expressed human alpha-1 antitrypsin for at least 2 months. In addition, we found that the hESC-derived hepatic cells were readily infected by human immunodeficiency virus-hepatitis C virus pseudotype viruses. CONCLUSION: We have developed an efficient way to direct the differentiation of human embryonic stem cells into cells that exhibit characteristics of mature hepatocytes. Our studies should facilitate searching the molecular mechanisms underlying human liver development, and form the basis for hepatocyte transplantation and drug tests. PMID- 17464997 TI - Bone marrow fails to differentiate into liver epithelium during murine development and regeneration. AB - Recent reports have provided conflicting conclusions regarding the role for bone marrow (BM)-derived cells in the regeneration of liver. Our aim was to investigate the potential of BM to contribute to liver epithelium using different BM transplant models designed to explore differentiation during normal liver development and regeneration after toxic injury. BM cells from transgenic green fluorescent protein (GFP) mice were injected into neonatal and adult immunodeficient and neonatal immune-competent mice. Three distinct models of liver injury were employed to test the contribution of marrow to the regeneration of hepatocytes, cholangiocytes, and oval cells in immune-deficient adult animals after neonatal transplant. Immunohistochemistry was combined with flow cytometry (FACS) and reverse transcription (RT)-PCR to increase the sensitivity and specificity of the analyses. Although GFP+ marrow-derived cells were observed in the livers of all transplanted animals, immunohistochemistry failed to demonstrate any marrow derived hepatocytes or cholangiocytes. FACS confirmed that GFP+ marrow-derived cells in the liver maintained expression of CD45, a leukocyte marker. Gene expression studies of GFP+ cells isolated by FACS failed to demonstrate expression of liver specific genes in these marrow-derived cells. CONCLUSION: Through highly sensitive and specific analyses, we were unable to demonstrate any evidence of transdifferentiation of BM-derived cells into epithelial hepatic tissue during the period of rapid growth in the neonatal period. Furthermore, although increased migration of hematopoietic cells to the liver occurred after toxic injury, these cells did not contribute directly to the replacement of hepatocytes, cholangiocytes, or oval cells. PMID- 17464998 TI - Incidence and risk factors for gallstones in patients with inflammatory bowel disease: a large case-control study. AB - The risk for gallstones (GD) in inflammatory bowel diseases and the factors responsible for this complication have not been well established. We studied the incidence of GD in a cohort of Crohn's disease (CD) and ulcerative colitis (UC) patients and investigated the related risk factors. A case-controlled study was carried out. The study population included 634 inflammatory bowel disease (IBD) patients (429 CD, 205 UC) and 634 age-matched, sex-matched, and body mass index (BMI)-matched controls free of GD at enrollment, who were followed for a mean of 7.2 years (range, 5-11 years). The incidence of GD was calculated by dividing the number of events per person-years of follow-up. Multivariate analysis was used to discriminate among the impact of different variables on the risk of developing GD. The incidence rates of GD were 14.35/1,000 persons/year in CD as compared with 7.75 in matched controls (P=0.012) and 7.48/1000 persons/year in UC patients as compared with 6.06 in matched-controls (P=0.38). Ileo-colonic CD location (OR, 2.14), disease duration>15 years (OR, 4.26), >3 clinical recurrences (OR, 8.07), ileal resection>30 cm (OR, 7.03), >3 hospitalizations (OR, 20.7), multiple TPN treatments (OR, 8.07), and long hospital stay (OR, 24.8) were significantly related to GD in CD patients. CONCLUSION: Only CD patients have a significantly higher risk of developing GD than well-matched hospital controls. Site of disease at diagnosis, lifetime surgery, extent of ileal resections, number of clinical recurrences, TPN, and the frequency and duration of hospitalizations are independently associated with GD. PMID- 17464999 TI - Increased activity of hepatic microsomal triglyceride transfer protein and bile acid synthesis in gallstone disease. AB - A strong interrelationship exists between the regulation of bile acid (BA) metabolism and hepatic very low density lipoprotein (VLDL) production. We have recently shown that BA synthesis is increased in gallstone disease. We investigated the activity of hepatic microsomal triglyceride transfer protein (MTTP) as a surrogate of VLDL production, BA synthesis, and mRNA expression levels of proteins that regulate fatty acid (FA) metabolism in the liver of gallstone (GS) patients compared with GS-free patients. Twenty-seven volunteers subjected to elective surgery; 9 were GS-free and 18 with GS agreed to have a liver biopsy. We quantified by a fluorescence assay the activity of MTTP and by quantitative reverse-transcription PCR (RT-PCR) the mRNA content of hepatic MTTP and genes that regulate hepatic sterol and FA metabolism. Plasma was assayed for lathosterol and 7alpha-hydroxy-4-cholesten-3-one. Liver histology was normal in GS and GS-free patients. Serum VLDL triglycerides and apoB were significantly increased in GS. Hepatic triglycerides tripled in GS (P<0.001) compared with GS free. MTTP activity increased 70% (P<0.001). Serum lathosterol and hepatic cholesterol concentrations, and mRNA expressions of MTTP, CD36, and FABP1 were similar in GS-free and GS patients. Hepatic mRNA expression of hydroxy-3 methylglutaryl-coenzyme A reductase (HMGR) and 3-hydroxyl-3-methyl-glutaryl-CoA synthase (HMGS) were significantly decreased--40% and 27%, respectively--in GS. Serum 7alpha-hydroxy-4-cholesten-3-one was 75% higher, and mRNA expression of CYP7A1 was increased sevenfold (P<0.001) in GS. CONCLUSION: Hepatic MTTP activity and BA synthesis are increased in GS. Results suggest that hepatic VLDL production and trafficking of BA are increased in gallstone patients. PMID- 17465000 TI - Cognitive function does not worsen during pegylated interferon and ribavirin retreatment of chronic hepatitis C. AB - Treatment of chronic hepatitis C with pegylated interferon (peginterferon) and ribavirin can cause or exacerbate depression but its effects on cognitive function are largely unknown. The aim of this study was to determine whether treatment with peginterferon and ribavirin adversely impacts cognitive function in patients with chronic hepatitis C. Prior nonresponders to interferon were retreated with peginterferon alfa-2a and ribavirin for 24 (n=177) or 48 weeks (n=57) in the Hepatitis C Antiviral Long-term Treatment Against Cirrhosis trial. Cognitive function was prospectively assessed using a battery of 10 standardized neuropsychological tests at weeks 0, 24, 48, and 72. Cognitive impairment was defined based upon a global deficit score. The Beck Depression Inventory and Brief Symptom Inventory were used to assess mood status. The 57 subjects who completed 48 weeks of antiviral therapy reported significant increases in difficulty concentrating, emotional distress, and symptoms of depression, all of which improved after cessation of therapy [P<0.0001, analysis of variance (ANOVA)]. Nonetheless, the frequency of cognitive impairment did not increase during the first 24 weeks of treatment in 177 patients (34% versus 32%, P=0.64) nor in the 57 patients completing 48 weeks of treatment (P=0.48, ANOVA). CONCLUSION: Retreatment of prior non-responders with peginterferon and ribavirin was not associated with objective evidence of cognitive impairment as measured by a comprehensive battery of neuropsychological tests. The lack of cognitive impairment is reassuring and suggests that self-reported symptoms of cognitive dysfunction are more likely related to the systemic and psychiatric side effects of antiviral treatment rather than measurable changes in cognition. PMID- 17465001 TI - The hepatitis C virus core protein of genotypes 3a and 1b downregulates insulin receptor substrate 1 through genotype-specific mechanisms. AB - Both molecular and clinical evidence support a link between HCV infection and insulin resistance. We examined the in vitro interaction between the HCV core protein of genotypes 3a and 1b with the insulin-signaling pathway. We measured the expression levels of insulin receptor substrate 1 (IRS-1), IRS-2, and other factors involved in the insulin signal pathway in a human hepatoma cell line (Huh 7) transiently expressing the HCV core protein of genotypes 3a or 1b by molecular biology and biochemical techniques. The IRS-1 (but not IRS-2) protein level was significantly reduced in Huh-7 expressing the core protein of both genotypes 3a and 1b, as compared to cells transfected with the empty vector. However, while the core protein of genotype 3a promoted IRS-1 degradation through the downregulation of peroxisome proliferator-activated receptor gamma (PPARgamma) and by upregulating the suppressor of cytokine signal 7 (SOCS-7), the core protein of genotype 1b activated the mammalian target of rapamycin (mTOR). We confirmed these findings by using agonists for PPARgamma (rosiglitazone) or short interfering RNAs for SOCS-7. CONCLUSION: Despite the small sequence divergence of the HCV core proteins of genotypes 3a and 1b, the 2 proteins appear to interfere with the insulin signaling pathway using genotype-specific mechanisms. PMID- 17465002 TI - Serum HBV RNA is a predictor of early emergence of the YMDD mutant in patients treated with lamivudine. AB - Lamivudine (LAM) is a nucleoside analogue widely used for the treatment of chronic hepatitis B virus (HBV) infection. Emergence of resistant strains with amino acid substitutions in the tyrosine-methionine-aspartate-aspartate (YMDD) motif of reverse transcriptase is a serious problem in patients on LAM therapy. The amount of covalently closed circular DNA in the serum is reported to be higher in patients who develop YMDD mutants than in those without mutants. However, there is no useful serum marker that can predict early emergence of mutants during LAM therapy. Analysis of patients who were treated with entecavir (n=7) and LAM (n=36) showed some patients had high serum levels of HBV RNA. Median serum levels of HBV RNA were significantly higher in patients in whom the YMDD mutant had emerged within 1 year (n=6, 1.688 log copies/ml) than in those in whom the YMDD mutant emerged more than 1 year after treatment (n=12, 0.456 log copies/ml, P=0.0125) or in whom the YMDD mutant never emerged (n=18, 0.688 log copies/ml, P=0.039). Our results suggest that HBV RNA is a valuable predictor of early occurrence of viral mutation during LAM therapy. PMID- 17465003 TI - HBsAg seroclearance in asymptomatic carriers of high endemic areas: appreciably high rates during a long-term follow-up. AB - Spontaneous hepatitis B surface antigen (HBsAg) seroclearance in chronic HBV infection has long been suggested as a rare event in high endemic areas. The prevalence of HBsAg in the general population of Taiwan, however, decreased remarkably from 15%-20% before age 40 to 5%-10% after age 60 or 70. This study aimed to reexamine the rates of HBsAg seroclearance by a long-term follow-up of 1965 hepatitis B e antibody-positive asymptomatic adult carriers. Of these, 1076 (55%) were males, the mean (+/-SD) age was 35.6+/-9.2 years and the mean follow up was 10.8+/-5.4 years. Hepatitis relapsed in 314 patients, 0.5 to 18 (mean+/ SD=5.8+/-4.4) years after the entry. The probability of hepatitis relapse correlated positively with male sex (P<0.0001) and age at entry (P<0.0001). Serum HBsAg cleared in 245 patients at the mean age of 47.8+/-9.6 years. The cumulative probabilities of HBsAg seroclearance were 8.1% after 10 years, but increased disproportionally to 24.9% and 44.7%, respectively, after 20 and 25 years. In multivariate analysis, the probability of HBsAg seroclearance correlated positively with age at entry (P<0.0001) and sustained remission of hepatitis (P<0.0001) and marginally significantly with male sex (P=0.053). CONCLUSION: Cumulative rate of HBsAg seroclearance in asymptomatic adult carriers from high endemic areas was approximately 40% after 25 years of follow-up. The low HBsAg seroclearance rates in previous studies might be due to the relative short period of follow-up. PMID- 17465004 TI - Impact of hepatitis B virus basic core promoter mutations on T cell response to an immunodominant HBx-derived epitope. AB - The hepatitis B X (HBx) protein is a crucial component in HBV infection in vivo and has been implicated in HCC. In this study, we aimed to detect and characterize peripheral HBx-specific T cells in chronically infected patients at the inactive carrier state of the disease. HBx-specific IFN-gamma-secreting T cells were found in 36 of 52 patients (69%), and 78% (28/36) of responding patients had T cells targeting epitopes in the carboxy-terminal part of HBx. IL 10 secretion after the stimulation of T cells with HBx-derived peptides was weak or undetectable. IFN-gamma-secreting T cells recognized a previously unknown immunodominant CD4+ T cell epitope, HBx 126-140 (EIRLKVFVLGGCRHK), in 86% (24 of 28) of patients. This peptide bound several HLA-DR molecules (HLA-DRB1*0101, HLA DRB1*0401, HLA-DRB1*1301, and HLA-DRB5*0101). Its coding sequence overlaps a domain of the HBV genome encompassing the basic core promoter (BCP) region. Taking into account the selection of viral core promoter mutants during HBV infection, we found that HBV variants with BCP mutations were present in patient sera. We further demonstrated that these viral mutant sequences activated T cells specific for the immunodominant epitope only weakly, if at all. This is the first study linking BCP mutations and HBx-specific T cell responses. CONCLUSION: Wild type and variant peptides may represent potential tools for monitoring the HBV specific T cell responses involved in sequence evolution during disease progression. Finally, the degenerate HLA-DR binding of this promiscuous, immunodominant peptide would make it a valuable component of vaccines for protecting large and ethnically diverse patient populations. PMID- 17465005 TI - Tim2 is expressed in mouse fetal hepatocytes and regulates their differentiation. AB - Liver development is regulated by various extracellular molecules such as cytokines and cell surface proteins. Although several such regulators have been identified, additional molecules are likely to be involved in liver development. To identify such molecules, we employed the signal sequence trap (SST) method to screen cDNAs encoding a secreted or membrane protein from fetal liver and obtained a number of clones. Among them, we found that T cell immunoglobulin and mucin domain 2 (Tim2) was expressed specifically on immature hepatocytes in the fetal liver. Tim2 has been shown to regulate immune responses, but its role in liver development had not been studied. We have examined the possible role of Tim2 in hepatocyte differentiation. At first, we prepared a soluble Tim2 fusion protein consisting of its extracellular domain and the Fc domain of human IgG (Tim2-hFc) and found that it bound to fetal and adult hepatocytes, suggesting that there are Tim2-binding molecules on hepatocytes. Second, Tim2-hFc inhibited the differentiation of hepatocytes in fetal liver primary culture, i.e., the expression of mature hepatic enzymes and accumulation of glycogen were severely reduced. Third, Tim2-hFc also inhibited proliferation of fetal hepatocytes. Fourth, down-regulation of Tim2 expression by small interfering RNA (siRNA) enhanced the expression of liver differentiation marker genes. CONCLUSION: It is strongly suggested that Tim2 is involved in the differentiation of fetal hepatocytes. PMID- 17465006 TI - Hepatic stellate cell protrusions couple platelet-derived growth factor-BB to chemotaxis. AB - Hepatic stellate cells play an essential role in the liver's injury response. Although stellate cells are defined by the presence of cytoplasmic protrusions, the function of these characteristic structures has been obscure. We hypothesized that stellate cell protrusions act by coupling injury-associated stimuli to chemotaxis. To test this hypothesis, we developed an assay for directly visualizing the response of living stellate cells in early primary culture to local stimulation of the tips of protrusions with platelet-derived growth factor BB (PDGF). Stellate cells exhibited elongate protrusions containing actin, myosin, and tubulin. PDGF, but not cytochrome C, localized at a protrusion tip induced a coordinated series of morphological events--cell spreading at the tip, movement of the cell body toward the PDGF, and retraction of trailing protrusions -that resulted in chemotaxis. Soluble PDGF and AG 1296, a receptor tyrosine kinase inhibitor, both reduced stellate cell chemotaxis. PDGF-induced chemotaxis was associated with an early and transient increase in myosin phosphorylation within the spreading lamella. We observed that blebbistatin, a myosin II inhibitor, completely and reversibly blocked protrusion-mediated lamella formation and chemotaxis. Moreover, blockade of MRLC phosphorylation with the myosin light chain kinase inhibitor, ML-7, or the rho kinase inhibitor, Y-27632, blocked lamella formation, myosin phosphorylation within the protrusion, and chemotaxis. CONCLUSION: These results support a model in which protrusions permit stellate cells to promptly detect PDGF distant from their cell bodies and transduce this signal into mechanical forces that propel the cell toward the site of injury. PMID- 17465007 TI - Sperm transfer and reproductive biology in species of hermaphroditic bivalves (Galeommatoidea: Montacutidae). AB - The histology of the reproductive organs is studied in the protandric hermaphroditic Tellimya ferruginosa. In NW Europe the species reproduces from May through August. Sperm transfer takes place when mature testis follicles are transplanted to the gills or walls of the mantle cavity in recipient hermaphroditic or female bivalves. Transplantation is accompanied by histological changes and sperm cells are released when transplants perish with age. Details are given on the reproduction in Montacuta percompressa which takes place from March through October in North Carolina, USA. All shelled bivalves are females and it is postulated that spermatogenic bodies attached to gills or other surfaces in the female's mantle cavity and previously considered to be dwarf males arise from transplanted larval gonads. The ultrastructure of the euspermatozoa and/or the anucleate paraspermatozoa is described in T. ferruginosa, T. tenella, and M. percompressa. The sperm of the first two species share a number of significant apomorphies with those of another montacutid, Brachiomya stigmatica. In the simultaneous hermaphroditic M. substriata the nucleate paraspermatozoa associate with the euspermatozoa to form spherical spermatozeugmata that are stored in the testis. PMID- 17465008 TI - Quantitative carbamylation as a stable isotopic labeling method for comparative proteomics. AB - A method was developed that uses urea to both solublize and isotopically label biological samples for comparative proteomics. This approach uses either light or heavy urea ((12)CH(4)(14)N(2)O or (13)CH(4)(15)N(2)O, respectively) at a concentration of 8 M and a pH of 7 to dissolve the samples prior to digestion. After the sample is digested using standard proteomic protocols and dried, isotopic labeling is completed by resuspending the sample in a solution of 8 M urea at a pH of 8.5, using the same isotopic species of urea as used for digestion and incubating for 4 h at 80 degrees C. Under these conditions, carbamylation occurs only on the primary amines of the peptides. The effects of complete carbamylation on matrix-assisted laser desorption/ionization time-of flight mass spectrometry (MALDI-TOFMS) and electrospray ionization tandem mass spectrometry (ESI-MS/MS) (collision-induced dissociation (CID)) were examined. Peptides that had a C-terminal carbamylated lysine residue were found to have a reduced intensity when viewed by MALDI-TOFMS. CID of a tryptic peptide that was carbamylated on both the N-terminus and the C-terminus was found to have a more uniform distribution of b- and y-ions, as well as prominent ions from loss of water. Reversed-phase chromatography coupled to ESI-MS/MS was used to identify and quantify the isotopically labeled standard proteins, bovine serum albumin (BSA), bovine transferrin, and bovine alpha-casein. Quantitative error between theoretical and observed data ranged from 1.7-10.0%. Relative standard deviations for protein quantitation ranged from 5.2-27.8% over a dynamic range from 0.1-10 (L/H). The development of a method utilizing urea-assisted carbamylation of lysines and N-termini to globally labeled samples for comparative proteomics may prove useful for samples that require a strong chaotrope prior to proteolysis. PMID- 17465010 TI - Determination of acidic degradation products of nonylphenol polyethoxylates by large-volume injection-port derivatization and gas chromatography/mass spectrometry. PMID- 17465009 TI - Fragmentation of 3-hydroxyflavone; a computational and mass spectrometric study. AB - In a recent study of the collision-induced dissociation of protonated and deprotonated molecules of 3-, 5-, 6-, 7-, 2'-, 3'- and 4'-hydroxyflavone, it was observed that the ratio, gamma, of the propensities for cross-ring cleavage (CRC) to ring opening (RO) varied by a factor of 660, i.e., from 0.014:1 (for deprotonated 3-hydroxyflavone) to 9.27:1 (for deprotonated 5-hydroxyflavone). An explanation for the variation of gamma was presented in terms of experimental NMR (13)C and (1)H spectra modified by computation. Deprotonated 3-hydroxyflavone exhibited the highest ion signal intensity for fragmentation following RO relative to that for CRC in that gamma = 0.014:1. Because no chemical computations of the fragmentation of protonated and deprotonated monohydroxyflavone molecules have been carried out thus far, the basis on which fragmentation mechanisms for deprotonated 3-hydroxyflavone have been proposed is principally chemical intuition. The energy states of product ions formed in the fragmentation of deprotonated 3-hydroxyflavone molecules were computed by the CBS 4M method implemented in Gaussian 03. The overly demanding calculations needed to handle diradicals reliably and directly were circumvented by a process in which each of the bond-breaking reactions was approximated by a process that gave two monoradicals. Bond energies were calculated, with one exception, from the approximation reactions as the energy of products minus the energy of reactants. PMID- 17465011 TI - Employing atmospheric pressure photoionization in liquid chromatography/tandem mass spectrometry to minimize ion suppression and matrix effects for the quantification of venlafaxine and O-desmethylvenlafaxine. AB - During the development of a method for quantitative determination of venlafaxine and its major metabolite O-desmethylvenlafaxine, elevated concentrations of the analyte as well as co-eluting matrix compounds caused ion suppression. This ion suppression was inconsistent and therefore influenced the reproducibility of detection. The use of atmospheric pressure photoionization (APPI) in the positive mode was investigated as a tool to circumvent this problem. Employing APPI resulted in negligible ion suppression and increased linearity of the concentration range. A selective, sensitive and rapid liquid chromatography/tandem mass spectrometry method for the determination of venlafaxine and its major metabolite O-desmethylvenlafaxine in human plasma was developed. The analyte was extracted from plasma into tert-butyl methyl ether followed by back extraction into 2% formic acid. An Agilent 1100 high-performance liquid chromatography (HPLC) system, employing reversed-phase chromatography on a cyano column, coupled to an Applied Biosystems API 3000 triple quadrupole mass spectrometer set to multiple reaction monitoring (MRM) mode, was used for separation and detection of the analytes. The method was validated between 2.36 605 ng per mL with a mean recovery of approximately 88% for both parent compound and metabolite analytes. APPI technology was employed to improve the reproducibility of detection enabling rapid, selective and sensitive quantification of venlafaxine and O-desmethylvenlafaxine in human plasma samples. PMID- 17465012 TI - Matrix-assisted laser desorption/ionization on-target method for the investigation of oligosaccharides and glycosylation sites in glycopeptides and glycoproteins. AB - The significance of glycoproteins in living systems instigates the ceaseless expansion of new techniques and procedures for the analysis of biological samples. Many of these applications are focused on improving the detection limit of analyzed material. In a previous study, we described a procedure for the detection of oligosaccharides cleaved from tryptic glycopeptides. Treatment of deglycosylated fractions with phenylhydrazine gave rise to peaks consistent with labeled glycans, and both types of compounds--deglycosylated peptides and oligosaccharides--were recorded from one spot and observed in one matrix-assisted laser desorption/ionization (MALDI) mass spectrum for the first time. Here, we added an additional step to this simple procedure of deglycosylating glycopeptides directly from the target spot of the first analyzed glycosylated peptides. For the purpose of this new study, a mixture of 2-aza-2-thiothymine and phenylhydrazine hydrochloride showed to be an excellent matrix for glycopeptides, oligosaccharides, deglycosylated peptides and moreover it allowed PNGaseF to be active enough to cleave oligosaccharides from peptides. The efficiency of this procedure is demonstrated on a series of intact glycoproteins and on the analysis of tryptic peptides obtained from IgG and total mouse serum. This one-step on target deglycosylation method with subsequent derivatization on the same spot makes MALDI-MS analyses of glycopeptides fast, simple and accessible for biological samples, where classical procedures cannot produce useful results. PMID- 17465013 TI - Denaturation of alpha-lactalbumin and ubiquitin studied by electrospray and laser spray. AB - Electrospray and laser spray mass spectra of human alpha-lactalbumin and bovine ubiquitin were studied, with an emphasis on the denaturation induced by laser spray. There were no remarkable differences in the electrospray and laser spray mass spectra for acidic and basic aqueous solutions of alpha-lactalbumin in positive and negative modes of operations. This originates from the fact that this protein is tightly folded with four disulfide bonds. For ubiquitin, however, denaturation was induced by laser spray for the positive mode of operation and the [M+nH](n+) with a maximum of n = 13 was observed, i.e., all the acidic amino acid residues are fully neutralized (protonated). In contrast, the laser-induced denaturation was not observed for the negative mode of operation, i.e., denaturation of ubiquitin is largely suppressed in the negatively charged liquid droplets. The marked difference observed in the positive and negative modes of operations for ubiquitin is ascribed to the difference in the susceptibility of side-chain/main-chain interactions in the positive-ion excess and in the negative ion excess liquid droplets. That is, the interactions between the basic residues and main-chain amide carbonyl groups (-NH(3) (+)***O=C< or -NH(2)***O=C<) which play an important role in stabilizing the protein structures are not so affected in the negative mode of operation but are weakened in the positive mode of operation. PMID- 17465015 TI - Application of polarity switching in the identification of the metabolites of RO9237. AB - Polarity switching mass spectrometry is an efficient way to collect structural data on drug metabolites. The value of this approach is illustrated with the in vitro metabolism of RO9237. Metabolites are identified by positive and negative electrospray ionization (ESI) full scan mass spectrometry, MS/MS and MS(3) using unlabelled and (14)C-radiolabelled versions of the drug. Comparison of the relative detectability of these metabolites by +ESI and -ESI shows that neither ESI mode is universal. It is advantageous to screen for metabolites using both positive and negative ionization modes. This is especially true for phase II metabolism which tends to make molecules more polar and often more acidic. Identification of phase II metabolites also benefits greatly from MS(3) experiments because the conjugating groups typically are cleaved in MS/MS and information on the core structure is only obtained in MS(3). A special case of phase II metabolism is the generation of glutathione (GSH) conjugates from reactive metabolites. The detection of GSH conjugates also benefits from generating both positive and negative ESI mass spectral data. PMID- 17465016 TI - Ergot alkaloids in Norwegian wild grasses: a mass spectrometric approach. AB - Ergot alkaloids are mycotoxins which are produced among fungi in the family Clavicipitaceae. Poisoning with ergot alkaloids is an important veterinary problem in animal husbandry and has recently also been recognised in wild animals. While the poisoning syndrome observed in domestic animals such as cattle, horses and sheep is usually caused by endophyte-infected grass, the recently observed ergotism among Norwegian cervids is probably due to infection of wild grasses with Claviceps. Mass spectrometry is today the method of choice for the rapid qualitative and quantitative determination of many natural compounds. This study uses tandem quadrupole mass spectrometry as well as ion trap mass spectrometry in connection with electrospray(+) ionisation for the quantification, screening and fragmentation of ergot alkaloids in extracts from Claviceps sclerotia that had been picked from wild grasses from several locations in Norway. Ergotamine, ergovaline, ergonovine and ergocryptine were available as standards and were quantified in the extracts, while ergocrystine, ergocornine, ergonine/ergosine, lysergic acid and lysergol were identified on the basis of their molecular weights and semi-quantified. Ergocrystine dominated the alkaloid spectrum of most extracts. Levels of the quantified alkaloids were in the range 0.2-9300 microg/g. Several unknown ergot alkaloids were found in the extracts. MS(n) experiments identified some as simple lysergic acid amide derivatives, while othes are probably related to ergocrystine and ergocryptine by dehydration, dehydrogenation and/or amino acid substitution at R(1) of the peptide moiety. PMID- 17465017 TI - Th17 and Th1 responses directed against the immunizing epitope, as opposed to secondary epitopes, dominate the autoimmune repertoire during relapses of experimental autoimmune encephalomyelitis. AB - Experimental autoimmune encephalomyelitis (EAE) is an inflammatory demyelinating disease with similarities to multiple sclerosis (MS). It has been suggested that relapses of EAE and MS may be associated with, and even driven by, T cells specific for novel epitopes that are primed during the course of tissue destruction in the target organ or in secondary lymphoid tissues. We show, however, that IFNgamma and IL-17 responses against the immunizing epitope remain dominant through out the course of multiphasic EAE. Furthermore, induction of tolerance against a putative secondary epitope did not prevent clinical relapses. PMID- 17465018 TI - Role of nitric oxide in the brain during lipopolysaccharide-evoked systemic inflammation. AB - Although the inducible isoform of nitric oxide synthase (iNOS) is a well established source of nitric oxide (NO*) during inflammation of the central nervous system (CNS), little is known about the involvement of constitutive isoforms of NOS (cNOS) in the inflammatory process. The aim of this study was to compare the responses of the expression and activity of iNOS and the two cNOS isoforms, neuronal and endothelial (nNOS and eNOS, respectively), in the brain to systemic inflammation and their roles in the cascade of events leading to degeneration and apoptosis. A systemic inflammatory response in C57BL/6 mice was induced by intraperitoneal injection of lipopolysaccharide [LPS; 1 mg/kg body weight (b.w.)]. The relative roles of the NOS isoforms were evaluated after injection of NG-nitro-L-arginine (NNLA; 30 mg/kg b.w.), which preferentially inhibits cNOS, or 1400W (5 mg/kg b.w.), an inhibitor of iNOS. Biochemical and morphological alterations were analyzed up to 48 hr after administration of LPS. Systemic LPS administration evoked significant ultrastructural alterations in brain capillary vessels, neuropils, and intracellular organelles of neurons, astrocytes, and microglia. Apoptotic/autophagic processes occurred in many neurons of the substantia nigra (SN), which coincided with exclusive enhancement of iNOS expression and activity in this brain region. Moreover, inhibitors of both iNOS and cNOS prevented LPS-evoked release of apoptosis-inducing factor (AIF) from SN mitochondria. Collectively, the results indicate that synthesis of NO* by both the inducible and constitutive NOS isoforms contribute to the activation of apoptotic pathways in the brain during systemic inflammation. PMID- 17465019 TI - Fabrication and in vitro characterization of three-dimensional organic/inorganic scaffolds by robocasting. AB - A key issue for the fabrication of scaffolds for tissue engineering is the development of processing techniques flexible enough to produce materials with a wide spectrum of solubility (bioresorption rates) and mechanical properties matching those of calcified tissues. These techniques must also have the capability of generating adequate porosity to further serve as a framework for cell penetration, new bone formation, and subsequent remodeling. In this study we show how hybrid organic/inorganic scaffolds with controlled microstructures can be built using robotic assisted deposition at room temperature. Polylactide or polycaprolactone scaffolds with pore sizes ranging between 200-500 microm and hydroxyapatite contents up to 70 wt % were fabricated. Compressive tests revealed an anisotropic behavior of the scaffolds, strongly dependant on their chemical composition. The inclusion of an inorganic component increased their stiffness but they were not brittle and could be easily machined even for ceramic contents up to 70 wt %. The mechanical properties of hybrid scaffolds did not degrade significantly after 20 days in simulated body fluid. However, the stiffness of pure polylactide scaffolds increased drastically due to polymer densification. Scaffolds containing bioactive glasses were also printed. After 20 days in simulated body fluid they developed an apatite layer on their surface. PMID- 17465020 TI - Functional characterization of missense variants in the creatine transporter gene (SLC6A8): improved diagnostic application. AB - Creatine transporter deficiency is an X-linked mental retardation disorder caused by mutations in the creatine transporter gene (SLC6A8). So far, 20 mutations in the SLC6A8 gene have been described. We have developed a diagnostic assay to test creatine uptake in fibroblasts. Additionally, we expanded the assay to characterize novel SLC6A8 missense variants. A total of 13 variants were introduced in the SLC6A8 cDNA by site-directed mutagenesis. All variants were transiently transfected in SLC6A8-deficient fibroblasts and tested for restoration of creatine uptake in deficient primary fibroblasts. Thus, we proved that nine variants (p.Gly87Arg, p.Phe107del, p.Tyr317X, p.Asn336del, p.Cys337Trp, p.Ile347del, p.Pro390Leu, p.Arg391Trp, and p.Pro554Leu) are pathogenic mutations and four variants (p.Lys4Arg, p.Gly26Arg, p.Met560Val, and p.Val629Ile) are nonpathogenic. The present study provides an improved diagnostic tool to classify sequence variants of unknown significance. PMID- 17465021 TI - Synthesis and characterization of radiopaque magnetic core-shell nanoparticles for X-ray imaging applications. AB - Radiopaque magnetic gamma-Fe(2)O(3)/poly(2-methacryloyloxyethyl(2,3,5 triiodobenzoate)) core-shell nanoparticles of narrow size distribution were prepared by emulsion polymerization of the iodinated monomer 2 methacryloyloxyethyl(2,3,5-triiodobenzoate) in the presence of maghemite (gamma Fe(2)O(3) nanoparticles coated with a dextran shell are commonly used as contrast agents for magnetic resonance imaging (MRI). The present nanoparticles have similar core-shell structure substituting the dextran for the iodo polymer. These core-shell nanoparticles may therefore be useful as imaging contrast agents to detect various pathogenic zones and to observe different disease states in both modes: X-ray and MRI. PMID- 17465022 TI - Characteristics of novel monofilament sutures prepared by conjugate spinning. AB - Compared with braided multifilament sutures, absorbable monofilaments are attractive suture materials as they exhibit less tissue drag and cause less tearing because of their smooth surfaces. However, monofilament sutures are less flexible and more difficult to tie a knot than multifilament ones, and their knots are more likely to loosen due to inferior knot security. Although various approaches have been reported to improve the flexibility of monofilament sutures, they still have limitations regarding poor knot security. To address this problem, we developed a novel technique to fabricate monofilament sutures by a conjugate spinning method, resulting in the formation of a sea/islands type of bicomponent monofilament suture. These sea/islands type bicomponent monofilament sutures, which can place many fine strands of a polymeric fiber within a matrix of another polymer, exhibited excellent knot security, flexibility, and low strain energy, compared with commercially available monofilament sutures. PMID- 17465023 TI - Development of an engineering autologous palatal mucosa-like tissue for potential clinical applications. AB - The goal of this study was to optimize key processes in recreating functional and viable palatal mucosa-like tissue that would be easy to handle and would promote wound healing. Normal human gingival fibroblasts and epithelial cells and a clinically useful biomaterial, CollaTape, were used. Structural and ultrastructural analyses showed that the gingival fibroblasts and epithelial cells adhered to the biomaterial and proliferated. Following a 6-day culture, using 10(5) fibroblasts and 10(6) epithelial cells, a well-organized palatal mucosa-like tissue was engineered. The engineered epithelium displayed various layers, including a stratum corneum, and contained cytokeratin 16-positive cells located in the supra-basal layer. This palatal mucosa-like engineered tissue was designed to meet a variety of surgical needs. The biodegradable collagen membrane (CollaTape) contributed to the flexibility of the engineered tissue. This engineered innovative tissue may contribute to the reconstruction of oral soft tissue defects secondary to trauma, congenital defects, and acquired diseases. PMID- 17465024 TI - Advantages of using glycolic acid as a retardant in a brushite forming cement. AB - In this study we have compared the effect of using acetic, glycolic, and citric acids on the brushite cement setting reaction and the properties of the resultant cement. The cement solid phase was made by mixing beta-tricalcium phosphate (beta TCP), monocalcium dihydrogen phosphate anhydrate (MCPA), and sodium pyrophosphate, whereas the cement liquid phase consisted of aqueous solutions of carboxy acids at concentrations ranging from 0.5 to 3.5M. Cements were prepared by mixing the solid phase with the liquid phase to form a workable paste. The cement setting time was longer for glycolic and citric acids. The best mechanical properties in dry environments were obtained using glycolic and citric acid liquid phases. In a wet environment at 37 degrees C, the cement set with glycolic acid was the strongest one. Brushite cement diametral tensile strength seems to be affected by the calcium-carboxyl phase produced in the setting reaction. The acceptable setting time and mechanical properties of cements set in glycolic acid solutions are attributed to the additional hydrophilic groups in the carboxylic acid and the low solubility in water of the calcium salt produced in the reaction. Moreover, at high concentrations, carboxylic acids add chemically to the cement matrix becoming reactants themselves. PMID- 17465025 TI - Repair of the dura mater with processed collagen devices. AB - BACKGROUND: We evaluated in a canine duraplasty model how specific differences in device physicomechanical properties, porosity, and crosslinking influenced the biological performance of three processed collagen dural substitutes. METHODS: Three collagen dural substitutes were studied: Dura-Guard, DuraGen, and Durepair. The initial strength, stiffness, and suture retention force were measured using standard mechanical test methods. The relative pore sizes of each device were assessed with a scanning electron microscope. Differential scanning calorimetry was used to measure their respective collagen denaturation temperatures. The biologic response and performance of the materials were evaluated via an acute (1 month) and long-term (3 and 6 months) canine bilateral duraplasty study. RESULTS: The mechanical properties of Dura-Guard and Durepair were similar to native dura. We could not quantify the mechanical properties of DuraGen because of its fragile nature. The denaturation temperature of DuraGen and Dura-Guard differed significantly from that reported for native collagens. The denaturation temperature of Durepair was comparable with the values reported for native collagens. All three materials were tolerated well by the animals. DuraGen did not maintain its structural integrity beyond 1 month. Dura-Guard and Durepair persisted for 6 months. Durepair was populated by fibroblasts and blood vessels, whereas Dura-Guard was not. CONCLUSIONS: The three dural substitutes tested were found to be safe and effective in healing surgically created defects in the dura mater. Although each of these dura substitutes are composed of collagen, differences in the collagen source and processing influenced device physicomechanical properties, porosity, and the nativity of the collagen polymer. These measured differences influenced device intraoperative handling and installation as well as the post-operative biological response, where differences in device resorption, cell penetration, vascularization, and collagen remodeling were observed. PMID- 17465026 TI - Acrylic injectable and self-curing formulations for the local release of bisphosphonates in bone tissue. AB - Two bisphosphonates (BPs), namely 1-hydroxy-2-[4-aminophenyl]ethane-1,1 diphosphonic acid (APBP) and 1-hydroxy-2-[3-indolyl]ethane-1,1-diphosphonic acid (IBP), have been synthesized and incorporated to acrylic injectable and self curing formulations. Alendronic acid monosodium trihydrated salt (ALN) containing cement was formulated as control. These systems have potential applications in low density hard tissues affected by ailments characterized by a high osteoclastic resorption, i.e. osteoporosis and osteolysis. Values of curing parameters of APBP and IBP were acceptable to obtain pastes with enough fluency to be injected through a biopsy needle into the bone cavity. Working times ranged between 8 and 15 min and maximum temperature was around 50 degrees C. Cured systems stored for a month in synthetic body fluid had compressive strengths between 90 and 96 MPa and modulus between 1.2 and 1.3 GPa, which suggest mechanical stabilization after setting and in the short time. BPs were released in PBS at an initial rate depending on the corresponding chemical structure in the order ALN > APBP > IBP to give final concentrations in PBS of 2.21, 0.44, and 0.19 mol/mL for ALN, APBP, and IBP, respectively. Cytotoxicities of bisphosphonates were evaluated, IC(50) values being in the order APBP > ALN > IBP. Absence of cytotoxicity coming from leachables of the cured systems was observed in all cases independently of the BP. An improved cell growth and proliferation for the systems loaded with APBP and IBP compared with that loaded with ALN was observed, as assessed by measuring cell adhesion and proliferation, and total DNA content. PMID- 17465027 TI - Development of a 95/5 poly(L-lactide-co-glycolide)/hydroxylapatite and beta tricalcium phosphate scaffold as bone replacement material via selective laser sintering. AB - 95/5 Poly(L-lactide-co-glycolide) was investigated for the role of a porous scaffold, using the selective laser sintering (SLS) fabrication process, with powder sizes of 50-125 and 125-250 microm. SLS parameters of laser power, laser scan speed, and part bed temperature were altered and the degree of sintering was assessed by scanning electron microscope. Composites of the 125-250 microm polymer with either hydroxylapatite or hydroxylapatite/beta-tricalcium phosphate (CAMCERAM II were sintered, and SLS settings using 40 wt % CAMCERAM II were optimized for further tests. Polymer thermal degradation during processing led to a reduction in number and weight averaged molecular weight of 9% and 12%, respectively. Compression tests using the optimized composite sintering parameters gave a Young's modulus, yield strength, and strain at 1% strain offset of 0.13 +/- 0.03 GPa, 12.06 +/- 2.53 MPa, and 11.39 +/- 2.60%, respectively. Porosity was found to be 46.5 +/- 1.39%. CT data was used to create an SLS model of a human fourth middle phalanx and a block with designed porosity was fabricated to illustrate the process capabilities. The results have shown that this composite and fabrication method has potential in the fabrication of porous scaffolds for bone tissue engineering. PMID- 17465028 TI - Gonadotropin-releasing hormone modulates voltage-activated sodium current and odor responses in Necturus maculosus olfactory sensory neurons. AB - The terminal nerve (nervus terminalis) extends from the basal forebrain to the nasal cavity and has been shown to contain gonadotropin-releasing hormone (GnRH). The specific function of the terminal nerve is unknown, but it has been hypothesized that it modulates the function of olfactory neurons. To examine the effects of GnRH on isolated Necturus maculosus olfactory sensory neurons (OSNs), we used the perforated configuration of the patch clamp technique to record current responses. GnRH had no effect on the membrane current at any holding potential but did modulate voltage-activated TTX-sensitive sodium current (INa). Within 1 min of applying GnRH, approximately 60% of the OSNs showed a decrease in the magnitude of INa. Initial responses to GnRH were inhibitory, although in one group of cells the initial inhibitory response was followed by a potentiation of INa with continual application (approximately 5 min). The time course of the GnRH response suggested that a second messenger pathway mediated the response. Inhibitors of PKC, tyrosine kinase, and PI3K were all able to inhibit the INa, but none of them could prevent the GnRH response. Application of a cAMP analog mimicked the effects of GnRH, and only inhibitors of PKA and PKG could prevent GnRH-induced inhibition of INa. This suggests that the modulation of voltage activated sodium currents by GnRH involve a cyclic nucleotide pathway. In addition, GnRH modulated the odor responses of OSNs. Our data suggest the release of GnRH, presumably from the terminal nerve, can serve to modulate olfactory sensory neurons. PMID- 17465029 TI - Accumulation of phosphorylated alpha-synuclein in dopaminergic neurons of transgenic mice that express human alpha-synuclein. AB - Parkinson's disease is neuropathologically characterized by the presence of Lewy bodies, whose major component is alpha-synuclein. We had previously generated transgenic mice that expressed human alpha-synuclein carrying an Ala53Thr point mutation (halpha-syn140m) under the control of the rat tyrosine hydroxylase promoter and found that halpha-syn140m was localized not only in the cytoplasm but also in the nuclei of mesencephalic dopaminergic neurons. In the present study, we carried out immunohistochemical analysis of the brain of Tg mice using anti-PSer129, an antibody that specifically recognizes alpha-synuclein phosphorylated at Ser129. The antibody detected only phosphorylated halpha syn140m, whereas phosphorylation of endogenous alpha-synuclein, if any, was below the detection limit of the method employed. The analysis showed that approximately one-third of the halpha-syn140m-positive neurons in the midbrain of heterozygous Tg mice were concomitantly reactive to anti-PSer129. The ratio almost doubled in homozygotes, indicating that the phosphorylation level depends directly on the amount of substrate. In addition, the ratio did not change at least up to 48 weeks of age. These data strongly suggest that halpha-syn140m underwent constitutive phosphorylation and that the phosphorylation level was maintained to a certain level until the aged stages. Remarkably, halpha-syn140m localized in the nuclei seemed to be preferentially phosphorylated compared with that in the cytoplasm. Among kinases that have been reported to be involved in the phosphorylation of alpha-synuclein, the beta subunit of casein kinase-2 was detected in the nuclei by immunohistochemistry. These data imply that at least casein kinase-2 is involved in the phosphorylation of halpha-syn140m in the Tg mice. PMID- 17465030 TI - Smooth muscle-associated protein 8: distribution and biological activity in the rat brain. AB - With the use of an antiserum directed against the human smooth muscle-associated protein 8 (SMAP8) fragment SMAP8(98-138), Western blot and immunohistochemical studies revealed SMAP8 expression in the rat brain. A band with a molecular size of about 45 kDa was detected in tissues from the rat hypothalamus and a weaker band from the cortex. SMAP8 immunoreactivity (irSMAP8) was detected in neurons of the hypothalamic paraventricular, supraoptic, and supraoptic retrochiasmatic nuclei; a few irSMAP8 cells were scattered in the zona incerta as well as the cerebral cortex. Immunoreactive cell processes were detected mostly in the internal layer of the median eminence. Double labeling the hypothalamic sections with SMAP8 and vasopressin (VP) or oxytocin (OT) antiserum revealed that a population of VP- and OT-immunoreactive neurons expressed irSMAP8. The biological activity of SMAP8 in rat central neurons was assessed by the calcium microfluorimetric Fura-2 method. SMAP8 (100 nM) elevated cytosolic calcium concentrations [Ca2+]i in a population of dissociated and cultured rat hypothalamic neurons; the response was eliminated in Ca2+-free saline. This is the first evidence of irSMAP8 in a population of VP/OT-containing hypothalamic neurons in the rat, and the peptide is biologically active in hypothalamic neurons, as evidenced by mobilization of extracellular Ca2+. PMID- 17465032 TI - Modafinil: serious skin reactions. PMID- 17465031 TI - Voluntary exercise-induced neurogenesis in the postischemic dentate gyrus is associated with spatial memory recovery from stroke. AB - Spatial cognitive impairment is common after stroke insults. Voluntary exercise could improve the impaired spatial memory. Newly generated neurons in the dentate gyrus are necessary for the acquisition of new hippocampus-dependent memories. However, it is not well known whether voluntary exercise after stroke promotes neurogenesis in the adult dentate gyrus, thereby promoting spatial memory recovery. Here, we examined in mice subjected to focal cerebral ischemia the effect of voluntary or forced exercise on neurogenesis in the ischemic dentate gyrus and spatial memory. Exposure to voluntary wheel running after stroke enhanced newborn cell survival and up-regulated the phosphorylation of cAMP response element binding protein (CREB) in the dentate gyrus and reversed ischemia-induced spatial memory impairment. However, the enhanced newborn cell survival and CREB phosphorylation in the dentate gyrus and improved spatial memory were not observed in the mice exposed to forced swimming. Moreover, there was a significant correlation between the total number of surviving newborn cells in the dentate gyrus and the ability of mice to locate the platform in the Morris water maze. These results suggest that, in the adult mice, exposure to voluntary exercise after ischemic stroke may promote newborn cells survival in the dentate gyrus by up-regulating CREB phosphorylation and consequently restore impaired hippocampus-dependent memory. PMID- 17465033 TI - Lamotrigine: birth defects. PMID- 17465034 TI - Telithromycin: QT prolongation. PMID- 17465035 TI - Intravenous iron and hypersensitivity. PMID- 17465036 TI - Imatinib: cardiac risks? PMID- 17465037 TI - Alendronic acid: synovitis, myalgia and arthralgia. PMID- 17465038 TI - Rotarix: injected by mistake. PMID- 17465039 TI - Reply to "Panico MG, Caporale V, Attena F. Adverse events following hexavalent vaccine (Hexavac): knowledge and information sources". PMID- 17465040 TI - Public reporting and pay for performance. PMID- 17465041 TI - Public reporting and pay for performance. PMID- 17465042 TI - Public reporting and pay for performance. PMID- 17465043 TI - Mortality after release from prison. PMID- 17465044 TI - Mortality after release from prison. PMID- 17465045 TI - Concussion. PMID- 17465046 TI - Concussion. PMID- 17465047 TI - Concussion. PMID- 17465048 TI - Concussion. PMID- 17465049 TI - The increase in solariums in Australia, 1992-2006. PMID- 17465050 TI - [An overview on stroke]. PMID- 17465051 TI - [Molecular mechanism of atherothrombosis]. PMID- 17465052 TI - [Apoptotic mechanism of ischemic brain damage]. PMID- 17465053 TI - [Mechanisms of ischemic cellular injury in response to reduction in cerebral blood flow]. PMID- 17465054 TI - [JSS-DE]. PMID- 17465055 TI - [National Institutes of Health Stroke Scale and Japan Stroke Scale]. PMID- 17465056 TI - [Brain infarction]. PMID- 17465057 TI - [Guideline for the brain imaging of acute ischemic stroke]. PMID- 17465058 TI - [The Guideline for Brain Dock 2003]. PMID- 17465059 TI - [Transcranial Doppler]. PMID- 17465060 TI - [Transesophageal echocardiography (TEE)]. PMID- 17465061 TI - [Magnetoencephalography (MEG) and transcranial magnetic stimulation (TMS)]. PMID- 17465062 TI - [Depression after stroke]. PMID- 17465063 TI - [Post stroke swallowing dysfunction and aspiration pneumonia]. PMID- 17465064 TI - [PICA study--hypertension and asymptomatic cerebral infarction]. PMID- 17465065 TI - [Pleiotropic effects of tissue plasminogen activator]. PMID- 17465066 TI - [Clopidogrel]. PMID- 17465067 TI - [Abciximab]. PMID- 17465068 TI - [Gene therapy for cerebrovascular disease]. PMID- 17465069 TI - [Cell replacement therapy by recruitment of endogenous neural progenitors]. PMID- 17465070 TI - [Regeneration therapy for ischemic stroke using neural stem cell transplantation]. PMID- 17465071 TI - Routine analgesia after episiotomy and perineal tear. PMID- 17465072 TI - A new "Alli" in weight loss. PMID- 17465073 TI - [Pollution by dust, smoke, gases and vapors]. PMID- 17465074 TI - Outbreak news. Meningococcal disease, Burkina Faso--update. PMID- 17465075 TI - Validation of neonatal tetanus elimination in India using lot quality-assurance cluster sampling. PMID- 17465076 TI - Africa Malaria Day. PMID- 17465077 TI - Price of second-line antiretroviral therapy reduced. PMID- 17465078 TI - Monitoring of neuraminidase inhibitor resistance among clinical influenza virus isolates in Japan during the 2003-2006 influenza seasons. PMID- 17465079 TI - Medical schools and teaching hospitals tag their economic impact at $451 billion. PMID- 17465080 TI - Functional biliary disorders: the Rome III experience. PMID- 17465081 TI - Antibiotics for acute necrotizing pancreatitis. PMID- 17465082 TI - [Autism, biomedical information and therapeutic alliance]. PMID- 17465083 TI - [CME-Ultrasonography 16/Solution]. PMID- 17465085 TI - Prevalence and economic implications of chronic pain. AB - Chronic pain is a major public health issue that affects the quality of life and productivity. It is costly and has a significant impact on health resource utilization. Management of chronic pain requires a multidisciplinary approach that focuses on disease management and takes into account the need for ongoing support by family members and other caregivers. Managed care pharmacies can play an important role in pain management to effect positive outcomes and reduce health resource utilization. PMID- 17465084 TI - Dependence of calcium on thyroid hormone for the regulation of cellular functions. AB - Recently, the thyroid hormone has been shown to cause increase in Ca2+ concentration by mobilizing intracellular Ca2+. The mobilization of intracellular Ca2+ in the absence of transmembrane Ca2+ influx has been accepted as evidence for a cell-surface Ca2+- receptor. The possible role of thyroid hormone in the regulation of cellular functions by Ca2+-channel was investigated using parameters proven to indispensably involve Ca2+: Bleeding and clotting times and rat uterine muscle activity in Ca2+-free Tyrode's solution. Adult female Wistar rats divided into four groups, namely; (i) Control, (ii) Thyroidectomized, (iii) Throidectomized-treated with thyroxine and (iv) Thyroxine-treated; were fed on rat chow. While groups i and ii were allowed access to tap water ad libitum for 60 days, groups iii and iv received tyroxine (10mg/kg body weight) every alternate day and tap water ad libitum for 60 days. All groups were kept in an air-conditioned room till they were euthanized. The results showed that thyroxine elicited biphasic contraction in Ca2+ - depletion and potentiated uterine contraction during Ca2+- loading (P<0.001). Inhibition of contractions in thyroidectomy, showed dependency of Ca2+ on thyroid hormones for the regulation of cellular functions. PMID- 17465086 TI - Treatment options for chronic pain management: opioids revisited. AB - Pain is a public health disaster. Although it remains largely undertreated, pharmacists and pharmacy managers can do much to correct the problem. Change will require the education of patients and health care professionals on the facts pertaining to addiction, drug dependence, and drug tolerance. Evidence-based resources, including guidelines published by the APS, should be consulted. A variety of short- and long-acting drugs are available, not the least of which are the opioids. Fears regarding addiction and uncertainty about tolerance have precluded the comprehensive use of these drugs, and varying dose requirements complicate the picture. What is known, however, is that dose titration must be performed with the goal of preventing breakthrough pain. Fortunately, sufficient drug formulae have been developed to accommodate this need. Pain is the primary reason why people come into the health care system, and it is incumbent upon health care professionals to eliminate insufficient pain control, using the drugs available to the patient's best advantage. PMID- 17465087 TI - Disease management tools for chronic pain. PMID- 17465088 TI - Case studies: managing debilitating chronic pain. PMID- 17465089 TI - Medical workforce planning and recruitment must support doctors. PMID- 17465090 TI - New drugs for the treatment of type 2 diabetes. AB - Insights into the pathogenesis of type 2 diabetes has led to increasing therapeutic targets for intervention, and improvements in peptide delivery technology have lead to non-injection methods of insulin delivery. This article reviews new drugs which are likely to become available for use by people with diabetes over the coming year. PMID- 17465091 TI - Diabetic ketoacidosis: principles of management. AB - Diabetic ketoacidosis has a mortality of up to 14% in the UK, which could be attributed to varied treatment policies and a lack of a general consensus statement on its management. This article provides comprehensive guidelines for management of adult patients in diabetic ketoacidosis. PMID- 17465092 TI - New-onset diabetes after transplantation. AB - New-onset diabetes after transplantation is a serious complication of organ transplantation that is becoming increasingly more common. New-onset diabetes after transplantation has implications for graft and patient survival. This article reviews the pathogenesis, diagnosis and management of new-onset diabetes after transplantation. PMID- 17465093 TI - Headache and migraine. AB - Headache is the commonest neurological presentation. Most patients have a benign pain disorder. The majority manage themselves with over-the-counter medications. Unfortunately therapeutic strategies in primary and secondary health care remain suboptimal and thus the condition poorly managed. PMID- 17465094 TI - Surviving shift systems: a guide for the foundation year doctor. PMID- 17465095 TI - The applied anatomy of examination of the knee. PMID- 17465096 TI - An introduction to fluid therapy. PMID- 17465097 TI - Survival kit for a foundation doctor. PMID- 17465098 TI - Injuries to the clavicle and acromioclavicular joint. PMID- 17465099 TI - Haemothorax. PMID- 17465100 TI - Advice for foundation doctors on dealing with elderly patients. PMID- 17465101 TI - Practical management of common atrial arrhy thmias 2: common atrial flutter. AB - This is the second in a series of four practical articles highlighting the important management steps for noncardiologists and non-cardiac electrophysiologists dealing with patients with atrial fibrillation and common atrial flutter. This article will deal with care pathways and management principles for common atrial flutter. PMID- 17465102 TI - Subcutaneous immunoglobulin for patients with antibody deficiency. AB - This article provides an overview on the management of primary immunodeficiency, and discusses the pharmacokinetics of subcutaneous and intravenous immunoglobulin and the relationship between blood levels and therapeutic effects in both treatments. The article will further highlight both treatments' efficacy in preventing infections and the risk of systemic side effects with each treatment. PMID- 17465103 TI - Career choices: foundation year trainees' views on careers advice and information. AB - This article looks at careers information sources and advice for junior doctors in foundation training programmes and their specialty of choice. The need for targeted career planning is discussed. PMID- 17465104 TI - Spontaneous uterine artery rupture in pregnancy. PMID- 17465105 TI - Digoxin toxicity in renal failure: role of the electrocardiogram. PMID- 17465106 TI - Computed tomography visualization of intracerebral haematoma 'pouring' directly into ventricular system. PMID- 17465107 TI - Immunohistochemical characterization of primary urinary bladder endometriosis. PMID- 17465108 TI - Is there a doctor on board? PMID- 17465109 TI - Debt is a family affair. PMID- 17465110 TI - Methadone-associated QT prolongation and torsades de pointes. PMID- 17465111 TI - Should hospitals have a medical emergency team? PMID- 17465112 TI - The red tide event in El Salvador, August 2001-January 2002. AB - A red tide event occurred in El Salvador from August 2001 to January 2002. National health authorities usually measured toxin levels in Ostrea iridescens, however other species were analyzed during this microalgae bloom: Anadara similis, Anadara tuberculosa and Modiolus sp. El Salvador authorities consider 400 mouse units/100 g the highest value that is safe for human health. During this period toxin levels in 0. iridescens and Modiolus sp. increased from values under 400 to 3977 and 15,468 mouse units/100 g, respectively. Persistent and higher levels were recorded in oyster and mussel banks on the west part of the country. The Ministry of Health and Social Assistance treated 41 slight to moderate intoxications associated to bivalve mollusks consumption. PMID- 17465113 TI - [Potentially toxic and harmful phytoflagellates from the Mexican Pacific coasts]. AB - The phytoflagellates are a heterogeneous group of autotrophic, heterotrophic and mixothrophic flagellates of trophic importance in several ecosystems. As in the rest of Latin America, the phytoflagellates that occur in the Mexican Pacific coasts are virtually unknown except for a few records. Their study require complicated collection and analysis methods, a probable cause for the scarce knowledge of this group in tropical and subtropical areas. Material recently collected from various localities along the Mexican Pacific coasts was used to study phytoflagellates, including toxic and potentially toxic species. Plankton samples were treated by gravity and pump filtration, using different methods for fixation and analysis. The phyla Euglenophyta, Heterokontophyta and Haptophyta were found. They occur as plankton in oceanic and shallow coastal waters. PMID- 17465114 TI - The use of pigment "fingerprints" in the study of harmful algal blooms. AB - Along the Mexican coast, harmful algae blooms (HAB) have become more frequent, and therefore, there is an urgent need to establish monitoring programs to avoid the undesired consequences of HAB in human and natural ecosystems. In this work, we analyzed the pigment signatures and the species composition from phytoplankton samples to evaluate the utility of the specific pigment "fingerprints" in HAB monitoring programs. Vertical profiles from a coastal lagoon and temporal samples of a red tide occurring in a shrimp-culture pond and in a coastal zone were taken into consideration. Between 76% and 84% of dinoflagellate and diatom cell density was explained by their specific signature variation, in both vertical and temporal samples. Only the variation of zeaxanthin and the cyanobacteria Anabaena sp. showed a poor relationship, probably from difficulties in counting other cyanobacteria present in the samples examined with the microscopic method. These results suggest that inclusion of pigment analysis in the study and monitoring programs dealing with harmful algae would be very useful. PMID- 17465115 TI - [Discoloration of microalgal blooms as a bioindicating parameter in the Bahia de Mazatlan, Mexico]. AB - In order to verify whether harmful algal blooms increased since 1979, a long-term project for the study of harmful algal blooms in Mazatlan Bay began 25 years ago (1979-2003). Observations were done daily and results indicate five trend types: (I) years without discoloration related to El Nino events; (II) years with days of sudden occurrence of discoloration, related to the release of accumulated energy after El Nino events; (III) years with similar number of discoloration events (days); (IV) years with a progressive decrease of the occurrence (days) of discoloration events; (V) years with gradual increase of discoloration events. The annual mean average of discoloration is of 25.3 days. Ninety-six discoloration events were recorded during year 2000, and represent the highest number of events in the last 25 years. Discoloration events are more common in March (174 days), harmful and toxic blooms included. This indicates continuity in the occurrence of the highest content of microalgae, and therefore, it is concluded that they play an important role for a better understanding of the environmental deterioration undergone by the coastal zone due to an increase of harmful and toxic microalgae blooms. PMID- 17465116 TI - [Presence of Cochlodinium catenatum (Gymnodiniales: Gymnodiniaceae) in red tides of Bahia de Banderas, Mexican Pacific]. AB - The evolution of an ichthiotoxic algal bloom caused by the dinoflagellate Cochlodinium catenatum was studied from July to December 2000. The abnormal multiplication of this dinoflagellate occurred in the form of a discoloration spreading between a temperature and salinity interval of 25-32 degrees C and 33 35 ups, respectively. The density of C. catenatum reached 10 841 cells ml(-1). The event was observed in large areas of Banderas Bay affecting 13 fish species, whose massive killing was due to suffocation (gill obstruction and excessive mucus production). The human population around the area did not present respiratory affections or skin irritation. The C. catenatum measurements suggest a hologamic and heterothalic reproduction. Their morphological characteristics suggest that C. polykrikoides, C. heterolobatum and C. catenatum are the same species. It is estimated that the species could be a recent introduction in the Mexican Pacific. PMID- 17465117 TI - Blooms of Cochlodinium polykrikoides (Gymnodiniaceae) in the Gulf of California, Mexico. AB - Cochlodinium polykrikoides was the species responsible for the discoloration that occurred between September 15th and 27th, 2000 in a shallow coastal lagoon located in the southern part of the Bahia de La Paz, on the west side of the Gulf of California. Blooms of C. polykrikoides were observed four days after two rainy days with a seawater temperature of 29 to 31 degrees C. Nutrient concentration ranges during the bloom were 0.165-0.897 microM NO2+NO3, 0.16-3.25 microM PO4, and 1.0-35.36 microM SiO4. Abundance of C. polykrikoides ranged from 360 x 10(3) to 7.05 x 10(6)/cells l(-1). Biomass expressed in terms of chlorophyll a was high, ranging from 2.7 to 56.8 mg/m3. A typical dinoflagellate pigment profile (chlorophyll a and c, peridinin, diadinoxantin, and beta-carotene) was recorded. In this study, the red tide occurred in front of several fish and shrimp-culture ponds. No PST toxins were found in the samples. However, 180 fish were found dead in the infected fish-pond; the gills were the most affected part. C. polykrikoides is a cyst-forming species that recurs in this area. New blooms were observed in November 2000 and September-November 2001 in the same area. Anthropogenic activities, such as eutrophication caused by water discharge in this shallow lagoon, and nutrient enrichment in the culture ponds, as well as effects from precipitation and wind stress, could have favored the outbreak of this dinoflagellate. PMID- 17465118 TI - [The red tide caused by the dinoflagellate Alexandrium tamarense in the Colombian Pacific coast (2001)]. AB - From April 26th to May 15th 2001, a large algae bloom was observed off Tumaco Bay on the Pacific coast of Colombia. This was the first harmful algae bloom (HAB) reported in the region, and reached Gorgona Island, about 120 km north. A year later, starting March 2002, an offshore HAB developed from Cabo Corrientes North to Solano Bay. The typical abundance during the blooms reached 7.5 x 10(6) cells l(-1) for the 2001 event and 1.6 x 10(6) cells l(-1) for the 2002 event. During both events, low temperature and high salinity were recorded. Typical measurements in the area are 27-27.5 degrees C and 30-31.5 psu. Values observed during the two events were 24-24.6 degrees C and 33-34 psu; 3 degrees C below normal and more than 2.5 psu above average values. These conditions are indicative of local upwelling processes at the time of the events. On both occasions, cells corresponding to the Alexandrium catenella/fundeyense/tamarense complex represented 99-100% of the biomass. It was difficult to differentiate the cells from A. catenella, but the presence of short chains of only 4 cells (single cells represented most of the biomass) was suggestive of A. tamarense. Shape, dimensions, and detailed structure of the apical pore complex, first apical plate, posterior sulcal plate, and position of the ventral pore on plate 1' of cells were consistent with the description of A. tamarense, which has not been reported in the tropical East Pacific. The Control Center of Pacific Contamination of the Maritime General Direction of the Colombian Navy has been monitoring the area since 1994 without finding this species or HABs. This leads us to consider the two events as caused by recently introduced species, where local upwelling processes favor permanent and cyclic HABs. However, during these two events, there were no reports of effects on marine biota or of human poisoning, probably because the blooms occurred some distance offshore and far from exploited shellfish beds. PMID- 17465119 TI - [Blooms of Pseudo-nitzschia spp. (Bacillariophyceae) and other phytoplankton species at Bahia de Mazatlan, Mexico]. AB - Observations on microplankton in the autumn of 1996 revealed the existence of high densities of Pseudo-nitzschia spp. A similar phenomenon was found in November 1998 and November 1999 and a month later it shifted to the southern Mazatlan Bay while in California, USA, incidents of the poisoning of sea mammals were being recorded; this also happened on the Mexican Pacific coasts from January to March. Water samples collected at surface (<0.5 m deep) and subsurface (10 m deep) monthly, from November 1998 to January 1999 and every week from February 1999 to May 2000, were analyzed by the Utermohl technique at 400 enlargements. Phytoplankton quantification showed winter and summer pulses; the 1998-1999 period was higher than the 1999-2000 period. Five species of Pseudo nitzschia were identified by electron microscopy (TEM and SEM); a new study has revealed other species. High values of Pseudo-nitzschia were observed: 730 P-n/ml (Dec. 1999), 610 P-n/ml (Dec. 1998), 335 P-n/ml (Jul. 1999), 198 P-n/ml (Feb. 1999), 170 P-n/ml (Nov. 1998), 123 P-n/ml (Mar. 1999), 108 P-n/ml (Aug. 1999). The blooms of other species of diatoms (Thalassiosira spp., Asterionellopsis glacialis, Chaetoceros spp., Skeletonema costatum), dinoflagellates (Prorocentrum spp., Scripsiella trochoidea) and ciliates (Myrionecta rubra) were also observed. PMID- 17465120 TI - [Species of dinoflagellates of the genus Gambierdiscus (Dinophyceae) in the Mexican Caribbean Sea]. AB - Some dinoflagellates with benthic habits are related to ciguatera intoxication by fish consumption, especially in tropical areas. In the Mexican Caribbean, ciguatera is relatively common, but only one paper seems to have been published on the subject, and there are very few publicactions on phytoplankton and benthic microalgae. Material collected along the coast of the State of Quintana Roo with phytoplankton net (54 mm) and directly from sediment and epiphytes of macroscopic plants, was searched for toxic and other associated dinoflagellates. Samples were studied by light and scanning electron microscopy. Morphological characters were useful for species identification, but eventually physiological, ecological and molecular characters could also be used. Three species of Gambierdiscus, related to the production of ciguatera toxins, were identified: G. belizeanus, G. toxicus and G. yasumotoi. They are distributed in shallow coastal areas, including coastal lagoons. PMID- 17465121 TI - Paralytic shellfish poisoning (PSP) in Margarita Island, Venezuela. AB - A severe outbreak of Paralytic Shellfish Poisoning (PSP) occurred in Manzanillo and Guayacan, northwestern coast of Margarita Island, Venezuela, between August and October 1991. A bloom of dinoflagellates including Prorocentrum gracile, Gymnodinium catenatum and Alexandrium tamarense seemed to be responsible for this outbreak. Levels of PSP toxins in mussels (Perna perna) exceeded the international safety limit of saxitoxin, 80 microg STX/100 microg meat. PSP toxin values varied between 2548 and 115 microg STX/100 g meat in Manzanillo, and between 1422 and 86 microg STX/100 g meat in Guayacan. At both locations, the highest levels were detected in August, when 24 patients exhibited typical symptoms of PSP toxicity after consuming cooked mussels (16 required hospitalization). A high pressure liquid chromatographic (HPLC) procedure was recently used on the 1991 samples. The major toxin detected in samples of both locations was decarbamoyl saxitoxin (dcSTX), but low concentrations of saxitoxin were also found in Manzanillo samples. Gonyautoxins GTX1, GTX2 and GTX3 were detected only at Guayacan, while in both locations, decarbamoylgonyatouxin (dcGTX2,3) toxins were detected. These findings represent the first time that causative toxins of PSP in Venezuela have been chemically identified, and confirm the presence of dcSTX and dcGTX in mussels from the Caribbean Sea. The presence of dcSTX and dcGTX in shellfish is indicative that Gymnodinium catenatum was a causative organism for outbreak of PSP. PMID- 17465122 TI - [Spatial-temporal dynamics of red tide precursor organisms at the Pacific coast of North and Central America]. AB - The Pacific coast of Central and North America has long been and still is impacted by the flourishing of microalgal populations known as Harmful Algal Blooms (HABs). The organisms that have caused recent HABs episodes in the region are among others, Gymnodinium catenatum, Pyrodinium bahamense var. compressum, and recently Cochlodinium cf. catenatum. In spite of the accumulated effects on the human health, the economic activities and the environment, scarce information is available on the subject. The augmented use of coastal zones for human activities is also paralleled by increased awareness of global climate changes. Thus, it is not an easy task to discriminate anthropogenic or natural phenomena, or both, as the major driving forces. The long-term data sets available for limited regions, as well as some sporadic observations during notorious blooms, allowed us to discriminate major changes in the biodiversity and biogeography of HAB organisms. Main changes refer to number of events, covered area, duration and frequency, number of blooming species and appearance of not previously reported harmful taxa. The variables more clearly related to these dynamic phenomena, seems to be sea surface temperature and wind force, but it is not yet possible to weight their contributions. The participation of rain is not fully evaluated to date. The collaborative communication among small-budget monitoring operations in the region allowed to "pass the voice" about peaking concentrations of HAB organisms, diminishing the risk of poisoning. PMID- 17465123 TI - Caribbean ciguatera: a changing paradigm. AB - Analyses of ciguatoxicity in the great barracuda Sphyraena barracuda and quantity of toxic benthic dinoflagellates on coastal reefs (correlated with the number of cases of human ciguatera intoxications in Puerto Rico) were used to construct a model formulated on data obtained during the period of 1985-1988. The validity of the proposed model has been questioned by recent data obtained during the period of 1990-2000. Barracuda ciguatoxicity no longer showed a prominent seasonality while the fraction of randomly caught barracuda that were ciguatoxic significantly increased during this period. These two changes, accompanied by the discovery that ciguatoxic fish contained a variety of multiple toxins, appear to be correlated with the steadily increasing periods of elevated sea surface temperatures in this region. PMID- 17465124 TI - [Presence of the dinoflagellates Ceratium dens, C. fusus and C. furca (Gonyaulacales: Ceratiaceae) in Golfo de Nicoya, Costa Rica]. AB - Harmful Algae Blooms (HAB) are a frequent phenomenon in the Gulf of Nicoya, Costa Rica, as in other parts of the world. The morphology and physiology of these microalgae are important because HAB species have adaptive characteristics. The production of high concentrations of paralytic toxins by Ceratium dinoflagellates has only been documented at the experimental level. However, this genus has been associated with the mortality of aquatic organisms, including oyster and shrimp larva, and fish, and with decreased water quality. Recently, fishermen reported massive mortality of encaged fish near Tortuga Island (Gulf of Nicoya). Samples were taken from an algal bloom that had produced an orange coloration and had a strong foul-smelling odor. Ultrastructural details were examined with scanning electron microscopy. The dinoflagellates Ceratium dens, C. furca and C. fusus were found in samples taken at the surface. The cell count revealed four million cells of this genus per liter. The morphological variability of these species is high; therefore electron microscopy is an useful tool in the ultrastructural study of these organisms. This is the first time that three Ceratium species are reported concurrently producing harmful blooms in Costa Rica. PMID- 17465125 TI - [Harmful blooms of cyanobacteria (Oscillatoriaceae) and dinoflagellates (Gymnodiniaceae) in the Golfo de Nicoya, Costa Rica]. AB - Recently, the Pacific coast of Costa Rica has experienced an increase in both magnitude and frequency of harmful algae blooms (HAB). The lack of data regarding the dynamics of these events in the area, and the species of microalgae that produce them, are themes of great interest. The blooms have produced negative impacts on fishery resources and on human health in Costa Rica. In May 2002 a HAB left a large number of dead fish along the central Pacific coast. Water samples were collected using a phytoplankton net and fixed for subsequent processing by electron microscopy. In addition, a one liter sample of surface water was taken for later cell count. In the observed HAB, the dominating organisms found were the cyanobacteria Trichodesmiun erythraeum surrounded by high concentrations of Gram-bacteria and the dinoflagellate Cochlodinium cf. polykrikoides. T. erythraeum, is one of the most important N2 fixing cyanobacteria in marine waters that has been associated with HAB events in diverse parts of the world as well as with symptoms that produce contact dermatitis and other discomforts. C. cf. polykrikoides is a dinoflagellete associated with fish kills; although the type of associated toxins are unknown. In a national newspaper 17 cases of intoxication in humans were reported during this same period, which presented respiratory disorders and burning of the eyes. This is the first report in Costa Rica where a cyanobacteria and a dinoflagellate were observed together producing HAB. PMID- 17465126 TI - [Algal blooms of the toxigenic diatom Pseudo-Nitzschia (Bacillariophyceae) in the Golfo de Nicoya, Costa Rica]. AB - Water samples were collected during a red tide event in November 2001, near San Lucas Island (Gulf of Nicoya, Costa Rica). Superficial temperature was 27 degrees C and water was turbid, with no fetid smell. One sample was treated with negative staining and observed using a transmission electron microscope (TEM); another sample was observed using a scanning electron microscope (SEM). Samples had high concentrations of the diatom Pseudo-Nitzschia pungensf pungens (characterized by two rows of poroids in the external channel), and lower concentrations of Skeletonema costatum (chains joined by external microtubules) and Chaetoceros lorenzianus (oval apertures and long chains, having setae with distinctive transverse rows and spines). This is the first time that the first species was described producing red tides in Costa Rica. However, reports about red tides with high concentration of species like P. pungens (variety multiseries) are increasing. These species have been related to the production of domoic acid, a low molecular weight amino acid which in humans can cause amnesic intoxications with seafood. Previously, Costa Rican reports of toxic accidents only referred to seafood contaminated with Pyrodinium bahamense var. compressum and Gymnodinium catenatum dinoflagellates. The increase in the number of Pseudo-Nitzschia causing harmful algae blooms is of interest for scientists around the world and must be documented. Similarly, some Chaetoceros species have been reported to be harmful to fish. We strongly recommend the establishment of a permanent surveillance program monitoring the presence of these species new at Costa Rican Pacific coast. Since the amnesic toxin is soluble in water and heat-resistant, we want to stress the possibility of having human cases of amnesic intoxication. PMID- 17465127 TI - Paralytic shellfish toxins in the chocolata clam, Megapitaria squalida (Bivalvia: Veneridae), in Bahia de La Paz, Gulf of California. AB - Occurrence and toxic profiles of paralytic shellfish toxins (PST) in the chocolata clam Megapitaria squalida were investigated. From December 2001 to December 2002, 25 clams were obtained monthly from Bahia de La Paz, Gulf of California. Additionally, net (20 microm) and bottle phytoplankton samples were also collected to identify toxic species. Toxins were analyzed by HPLC with post column oxidation and fluorescence detection. Toxicity in the clam was low and varied from 0.14 to 5.46 microg/STXeq/100 g. Toxicity was detected in December, March, April, June, and August. Toxin profile was composed mainly by STX, GTX2, GTX3, dcGTX2, dcGTX3, C2, dcSTX and B1. Gymnodinium catenatum was the only PST producing dinoflagellate identified in the phytoplankton samples throughout the study period. G. catenatum was observed mainly in net samples from December 2001 to December 2002; however, in bottle samples, G. catenatum was only observed in five months. Highest abundance (2600 cells l(-1)) was observed in March and the lowest (160 cells l(-1)) in June. G. catenatum mainly formed two-cell chains and rarely four or eight. The presence of PST in net phytoplankton samples support the fact that G. catenatum is the main source of PST in the clams. This study represents the first report of PST toxins in the chocolata clam from Bahia de La Paz. PMID- 17465128 TI - PCB concentrations in sediments from the Gulf of Nicoya estuary, Pacific coast of Costa Rica. AB - Thirty-one sediment samples collected from 1996-2003 from the Gulf of Nicoya estuary on the northwestern coast of Costa Rica, have been obtained for PCB analyses. This is part of the first study to evaluate the PCB contamination in coastal Costa Rica. Overall, the concentrations are low, especially when compared to sediments from more temperate climates and/or sediments from more heavily industrialized areas. Values average less than 3 ng/g dw sediment, however, a few samples contained up to 7 ng/g dw sediment. Sediments with the highest concentrations were located in the Punta Morales area, where muds were sampled from among mangrove roots. The Puntarenas samples had surprisingly low PCB concentrations, likely due to their sandy lithology. The congener distribution within the majority of the samples showed signs of either recent sources or lack of degradation. However, a few sites, specifically some of the inter-gulf islands and more remote samples had congener distributions indicative of airborne contaminants and/or degradation. Considering the presence of airborne PCBs in the Gulf of Papagayo to the north, the lack of airborne PCBs and more varied congener distribution in the Gulf of Nicoya estuary was surprising. PMID- 17465129 TI - PCB contamination in marine sediments from Golfo Dulce, Pacific coast of Costa Rica. AB - Twenty-nine marine sediment samples collected from 1996 through 2002 from the Golfo Dulce embayment of Costa Rica were analyzed for PCB concentrations. The Esquinas River and Rincon Bay in the northern and western part of the gulf had relatively low overall concentrations of PCBs, with no samples having greater than 2.1 microg/g dw sediment. The Port of Golfito had the highest overall concentrations, ranging up to 15.7 microg/g dw sediment. These samples were also dominated by higher chlorinated congeners. Samples from the deeper (> 100 m) waters in the northern part of the gulf, as well as within the sediment plume from the Rio Coto Colorado had intermediate values. Within the Rio Coto Colorado sediment plume the concentrations did decrease with increasing depth and the congeners showed a shift towards less chlorinated congeners with depth. However, the deep northern basin had some of the highest PCB concentrations and the shift towards less chlorinated congeners was not apparent or significant. Whether the anoxic conditions that exist in the deep waters are capable of initiating dechlorination is still unknown. Overall, the data from Golfo Dulce show moderate PCB contamination, despite the pristine nature of the gulf and surrounding lands. PMID- 17465130 TI - [Physical-chemical features and determination of pesticides in the lagoon water in Gandoca, Limon, Costa Rica. ]. AB - Nutrients and chlorophylls concentrations, as well as salinity, temperature and Secchi disk depth were determined from November 1999 to April 2000, at three stations and two depths, at Gandoca lagoon, Gandoca-Manzanillo National Wildlife Refuge, Limon, Costa Rica. Salinity profiles indicated that the lagoon was a salt wedge estuary with a partially mixed region near the mouth. No processes of eutrophication were found. The distribution and abundance of nutrients and chlorophylls showed a slight influence of continental water and water circulation patterns in the lagoon. A preliminary study was done in order to analyze the presence of 20 organochlorated and organophosphorated pesticides along the Gandoca lagoon in February 2000. None of the pesticides were detected by the analysis of residues from liquid-liquid extractions. The absence of the pesticides may be due to the fact that they did not reach the lagoon or, if they did, they were washed away by the strong rains during the sampling period. PMID- 17465131 TI - [Oil pollution status expressed as the fraction of dissolved and dispersed petroleum hydrocarbons]. AB - Four coastal ecosystems with contrasting characteristics were sampled in Costa Rica (2000-2002). Oil pollution status, expressed as the fraction of dissolved/dispersed petroleum hydrocarbons related to chrysene equivalents, was determined by the molecular fluorescence analytical technique. A total of 130 water samples were taken, from the Caribbean (Moin Bay), and from the Pacific (Bahia Culebra, Gulf of Nicoya and Dulce Gulf). On one occasion, seven samples along the Puntarenas estuary were also analysed. In Moin the mean and standard deviation were 0.10 microg x L(-1) +/- 0.18 micro x L(-1), ranging from non detectable (nd) to 0.65 microg x L(-1). For the Pacific ecosystems the total range was from nd to 0.37 microg x L(-1). In Bahia Culebra no fluorescence signals were obtained. In the Gulf of Nicoya the mean and standard deviation were 0.04 microg x L(-1) +/- 0.09 microg x L(-1), from nd to 0.33 microg x L(-1). Values in Dulce Gulf were 0.05 microg x L(-1) +/- 0.11 microg x L(-1), from nd to 0.37 microg x L(-1). Along the Puntarenas estuary the range was 0.17 to 5.91 microg x L(-1), with a mean of 1.21 microg x L(-1) and a standard deviation of +/ 2.10 microg x L(-1). The four coastal ecosystems had concentrations below the 10 microg x L(-1) limit for polluted oceanic areas. The Puntarenas estuary reflects the influence of antropogenic activities from and around the City of Puntarenas. These levels are considered low for inshore waters. PMID- 17465132 TI - [Composition of marine sediment samples in the Costa Rica intertidal zones using X-Ray fluorescence analysis]. AB - Using an energy dispersive X-Ray fluorescence analysis, simultaneous evaluation of K, Ca, Ti, Cr, Mn, Ge, Ni, Cu, Zn, Br, Rb, Sr and Pb in 74 marine sediment samples from the Costa Rica intertidal zones was conducted. Samples were collected between June 1999 and December 2001, from Caribbean and Pacific beaches of Costa Rica. Calcium and iron showed the highest abundances and are indicators of the natural origin of the sediments. Calcium is associated with biogenic processes such as coral reefs near the sampling sites and iron indicates a terrigenous origin. In general, the beaches of the Caribbean and North Pacific regions showed the greatest concentration of calcium. This is indicative of the abundant reef structures near these beaches. The beaches of the Central and South Pacific show the greatest iron concentrations, indicating an important lithosphere contribution and/or little contribution of calcium carbonate due to the poor development of coralline structures near the sampling sites. Finally, the analyses did not show evidence of elements associated with anthropogenic pollution. Only a northern section of Puerto Viejo beach showed high concentrations of lead, zinc and titanium, perhaps associated with hydrothermal sources. PMID- 17465133 TI - [Water mass behavior of Dulce Gulf, Costa Rica, during El Nino (1997-1998)]. AB - Abstract: In order to understand the vertical distribution of planktonic organisms in the oceans, it is important to consider a number of factors. Including oceanic circulation patterns, the vertical structure of the water column and the vertical position of the thermocline. This study examines the hydrography of Golfo Dulce, a tropical fjord -like embayment in the Eastern Tropical Pacific, and the influence of the 1997-1998 ENSO on the system. Five sampling stations were selected and visited once a month for two years in order to obtain vertical profiles of temperature (degrees C), salinity (% per hundred) and dissolved oxygen (mg O2/l). Results show a strong vertical stratification of the water column associated to the morphology of Golfo Dulce's basin and to the effects of ENSO. Moreover, the existence of significant differences between the hydrographic characteristics of the inner and outer parts of the basin was detected. Finally, biannual events in the influx of cold, high salinity, and oxygen rich oceanic water masses into the basin were detected. These events might be related to the same forces driving the coastal upwelling system in the Gulf of Panama. PMID- 17465134 TI - Composition, abundance and distribution of macrozooplankton in Culebra Bay, Gulf of Papagayo, Pacific coast of Costa Rica and its value as bioindicator of pollution. AB - The abundance, distribution and composition of the macrozooplankton of Culebra Bay, Costa Rica (10 degrees 38' N - 85 degrees 40' W) were studied at four stations throughout the dry (February-May) and rainy (September-November) seasons of 2000. The samples were collected at two-week intervals using a 500 microm mesh net with a 0.5 m diameter opening. Copepods (23-31%) and ostracods (20-34%) were predominant throughout the year, followed by cladocerans (2.5-14%), zoea (6.6 9.5%), and siphonophores (2.5-7.2%). High densities of zooplankton were obtained in February and March with peak abundance on March 18. The lowest densities were observed on September 3 and November 5. Significant differences in abundances at each station were observed for the groups Acartia tonsa (Copepoda), Ctenophora, Medusae, Ostracoda, Zoea, and Amphipoda. Comparison of the dry and rainy seasons revealed significantly higher zooplankton abundance in the dry season and copepod domination of all stations; during the rainy season ostracods dominated the off shore areas. Zooplankton abundance and distribution are influenced by upwelling, which occurs during the dry season in Culebra Bay. PMID- 17465135 TI - Marine biodiversity of Costa Rica: class Polychaeta (Annelida). AB - Three hundred and thirty two species of polychaetes belonging to 50 families are reported for Costa Rica based on previously published reports. In a few families, available material was re-examined and appropriate changes in species identifications were made. Only three species, (Glycera oxycephala Ehlers, 1887; Hemipodia pustulata (Friedrich 1956); and Scolepis (Scolepis) squamata (Miller, 1806)) have been reported from the Caribbean coast of Costa Rica, and these three species have also been reported from the Pacific Coast. The families with more than 15 species are the Spionidae (26), Cirratulidae (24), Nereididae (21), Lumbrineridae (19) and Paraonidae (16). Despite extensive sampling on the Pacific coast over the last 25 years, the Costa Rican polychaete fauna, especially that of the Caribbean coast, remains poorly known. PMID- 17465136 TI - Box-Jenkins analysis for shark landings in Costa Rica. AB - Sharks are highly vulnerable to intense and prolonged fishery extraction. This article analyzes the data on shark landings from the artisan fishing fleet on Costa Rica's Pacific coast between 1988 and 1997. The data come from an invoicing system administered by the Costa Rican Fisheries Institute (Instituto Costarricense de la Pesca y Acuacultura, INCOPESCA). Pacific coast shark fishing during the period under study represented approximately 20% of the total national fisheries volume. According to data from the invoicing system, the Northern Pacific region was the most productive, reporting 58% of the shark catch nationwide. Within this region, shark fishing in Papagayo Gulf represented 91% and 53% of the landings by fishery region and nationwide, respectively. The mid sized and advanced (length of boat > 10 meters) artisan fishing fleets reported 96% of the shark catches in the zone. The study of shark fisheries in the Papagayo Gulf zone is crucial for an understanding of fishery dynamics for this resource at the national level. A monthly chronological series was constructed with the landings in the Papagayo Gulf zone, and a Univariate Box-Jenkins (UBJ) Model was estimated for first-order moving averages MA(1) with a seasonal component of the Yt = lamda(t-1) + gammaS12 + a(t) type. PMID- 17465137 TI - [Sighting of Stenella attenuata, the spotted dolphin, in Culebra Bay, Costa Rica, 1999-2000]. AB - Parallel to a zooplankton study (1999-2000) observations were made (from an inflatable boat), on the presence of dolphins along a transect (-8 km long) on the axis of Culebra Bay (24 km2), Gulf of Papagayo, Pacific coast of Costa Rica. Dolphins were found during 20 of the 31 boat surveys conducted. The only species of cetacean found in the bay was Stenella attenuata, the spotted dolphin. These sightings were more frequent during the rainy season, particularly during the month of May of both years. The presence of S. attenuata in Culebra Bay might be associated to the abundances of fish and mollusks (their presumed prey: for example, squids), as evidenced by fishery statistics available for this zone of the Pacific coast of Costa Rica. PMID- 17465138 TI - [The Purruja mangrove, Golfito, Costa Rica: a management model]. AB - The Purruja mangrove (Golfito, Costa Rica) has an estimated area of 70 ha. A socio-biological research was the ground to set initial goals to manage the resources and to identify the mangrove biological condition. Community participation and the local organization were key factors in developing an integrated model for the management of natural resources. Constant monitoring and institutional networks were the other two factors to manage the mangrove. The constant profesional support was a tool to facilitate the acomplishment of goals and to establish an institucional network to promote local group iniciatives for collaborative management of the Purruja mangrove. PMID- 17465139 TI - Development and field application of a molecular probe for the primary pathogen of the coral disease white plague type II. AB - One of the current problems in the field of coral disease research is that of tracking coral pathogens in the natural environment. A promising method to do this is by use of pathogen-specific molecular probes. However, this approach has been little used to date. We constructed, and validated in the laboratory, a fluorochrome-labeled molecular probe specific to Aurantimonas coralicida, the bacterial pathogen of the Caribbean coral disease white plague type II (WPIl). We then used the probe to test field samples of diseased coral tissue for the presence of this pathogen. Probe design was based on a unique subset (25 nucleotides) of the complete l6S rRNA gene sequence derived from a pure culture of the pathogen. The pathogen-specific probe was labeled with the fluorochrome GreenStar* FITC (fluorescein isothiocyanate, GeneDetect Ltd, New Zealand). As a control, we used the universal eubacterial probe EUB 338, labeled with a different fluorochrome (TRITC, tetra-methylrhodamine isothiocyanate). Both probes were applied to laboratory samples of pure cultures of bacteria, and field samples collected from the surface of the disease line of corals exhibiting signs of white plague (types I and II), healthy controls, and corals with an uncharacterized disease ("patchy necrosis"). All samples were analyzed using fluorescence in situ hybridization (FISH). We have determined that the probe is specific to our laboratory culture of the coral pathogen, and does not react with other bacterial species (the eubacterial probe does). The WPII pathogen was detected in association with diseased coral samples collected from coral colonies on reefs of the Bahamas (n= 9 samples) exhibiting signs of both WPI and WPII. Diseased (and healthy) tissue samples (n- 4) from corals exhibiting signs of "patchy necrosis" were also assayed. In this case the results were negative, indicating that the same pathogen is not involved in the two diseases. Incorporation and use of pathogen-specific probes can significantly expand our knowledge of the etiology of coral diseases. PMID- 17465140 TI - The distribution and abundance of wetland ichthyofauna, and exploitation of the fisheries in the Godineau Swamp, Trinidad--case study. AB - The Godineau (South Oropuche) Swamp (3171 ha) on the southwestern coast of Trinidad (10 degrees l315'N, 61degrees 30-32'W) is heavily impacted by anthropogenic activities including fishing, oil exploration, drainage manipulation and wetland clearance. To reduce the negative effects of these activities and to manage the swamp more sustainably, more quantitative information is needed on the ecology of the wetland and the activities that occur within it. This study focuses on the distribution of the fish resources and exploited fisheries as a basis for more informed management directives. Sampling was conducted during 2002, in April-May (for dry season) and July-September (for wet season sampling). Ichthyofauna was sampled both day and night using trammel nets and a push seine. Fishing activities were assessed using a questionnaire and informal discussions with fishers. The wetland supports over 29 species of fish distributed over the freshwater, estuarine and saline zones (n=1454). Species distribution is seasonal, with evidence of the wetland being used as a spawning ground. Species richness and species diversity (Shannon-Weiner Index) for the wetland ranged over 2-11 and 0.162-0.967, respectively, in the dry season and 2-7 and 0.036-0.903, respectively, in the wet season. Communities inhabiting the saline and estuarine zones of the wetland were dominated by a single species, Hexanematichthvs bonillai. Percent Similarity Indices were 41.8% for freshwater, 72.7% for estuarine and 79.8% for estuarine-saline communities. The commercial species accounted for 18% of total catch sampled and consisted of Centropomus undecimalis, Megalops atlanticus, Hoplosternum littorale, Hoplias malabaricus, Ophioscion punctatissimus and Macrodon ancyclodon. Full-time and part-time fishers, including recreational fishers, accounted for 14.3% and 85.7%, respectively of all fishers surveyed (n=56). A conservative estimate of the revenue earned directly at point of sale for fish and shellfish, is approximately US$43 000 per annum. Fishing is important in the area. Management directives taking into account the ecology of the target organisms are needed to allow these fishing practices to be continued in a sustainable manner. PMID- 17465141 TI - Chlorophyll a and turbidity patterns over coral reefs systems of La Parguera Natural Reserve, Puerto Rico. AB - Studies of temporal and spatial changes in phytoplankton biomass and turbidity provide essential information on coral reef ecosystem function and health. Fluctuation of phytoplankton biomass responds to several factors including nutrient inputs, both anthropogenic and natural, while turbidity is mostly affected by sediment resuspension or transport from terrestrial systems. These parameters can be used as sentinels of significant environmental factors "modifying" coral reef systems. A chlorophyll a concentration (Chl a) and turbidity (Turb) in situ logger was installed at 10 stations from June 4 to July 7, 2003 in La Parguera Natural Reserve (Southwestern Puerto Rico) to assess short term temporal and geographic variation in patterns of phytoplankton biomass and turbidity at pre-selected sites as part of an interdisciplinary long-term study. Average station Ch1 a variation was 0.17-1.12 microg 1(-1) and 0.2-23.4 NTU for Turb. Results indicate that the western near-coastal stations had higher levels of Turb and Ch1 a. The easternmost mid shelf station, Romero reef, was similar to coastal stations probably due to nutrient and suspended sediment inputs from a source external to our study area to the east, Guanica Bay. Comparisons between different sampling days indicate significant differences between days for most stations suggesting that one-time discrete sampling may not be representative of average water column conditions and illustrate the dynamic nature of coral reef systems. Further work is warranted to assess seasonal changes that integrate short-term (daily) variability in both Turb and Ch1 a. PMID- 17465142 TI - Spatial distribution of heavy metals in sediments from the Gulf of Paria, Trinidad. AB - The Gulf of Paria receives heavy metal input from urban runoff, industrial and agricultural activity, sewage and domestic wastes: both from the west coast and from inland areas of Trinidad. Non-residual concentrations of nine metals, as well as total mercury concentrations, were used to determine spatial distributions of heavy metals in sediments in the Gulf of Paria. Surficial sediment samples were collected at 37 stations, which included the mouths of 11 major rivers that flow into the Gulf of Paria. Stations were sampled twice during the wet season (July 1998 and November/ December 1998) and twice during the dry season (March 1999 and April 1999). Sediments were analyzed for aluminium (Al), cadmium (Cd), chromium (Cr), copper (Cu), iron (Fe), lead (Pb), manganese (Mn), nickel (Ni), zinc (Zn) and mercury (Hg). Total Organic Carbon (TOC) and grain size analyses were also performed on the sediments. Principal component analysis showed that sediments from river mouths subject to greatest land use and anthropogenic input, were distinct from other sediments in the Gulf of Paria. This was due to higher Pb, Zn, Cu and Hg concentrations (3.53-73.30 microg g(-1), 45.8-313.9 microg g(-1), 8.43-39.71 microg g(-1) and 0.03-0.10 microg g(-1), respectively). Sediments further from the coast were also distinct due to their higher Al, Fe, Cr and Mn concentrations (1.37-3.16 mg g(-1), 9.51-18.91 mg g(-1) , 17.22-28.41 microg g(-1) and 323.6-1,564.2 microg g(-1), respectively). Cd and Pb were higher in the wet season while Ni was higher in the dry season. Pb, Zn, Cu and Hg were correlated with each other and with TOC. Correlation was also observed between Al, Fe, Cr, Mn and Ni. Al, Fe, Cr and Mn were correlated with percentage clay in sediments. The results suggest that Pb, Zn, Cu and Hg are preferentially removed by organic matter, which settles at the river-mouths, while Al, Fe, Cr, Mn, and Ni become associated with clay minerals and are transported away from the coast. PMID- 17465143 TI - Heavy metals in sediments, mussels and oysters from Trinidad and Venezuela. AB - The Gulf of Paria is bordered by both Trinidad and Venezuela, from which various metallic pollutants and other contaminants can originate. The Gulf is still a significant source of fish, crabs and shellfish for human consumption to both countries, where concerns over the quality of this marine environment have been long expressed but never properly addressed. In addition, the circulatory current patterns in the Gulf ensure that contaminants originating from either country are likely to affect both countries eventually. Heavy metals were determined in oysters (Crassostrea rhizophorae and C. virginica), green mussels (Perna viridis) and sediments from the Gulf of Paria. Samples were obtained at four sites in Trinidad and three sites in Venezuela in the Gulf of Paria, in addition to comparative samples collected from three sites on the north coast of Venezuela. Edible tissues of twelve shellfish from each location were blended and aliquots digested with concentrated nitric acid, for extraction of cadmium, chromium, copper, lead, nickel and zinc. The solutions were analysed by flame atomic absorption spectroscopy. Mercury was extracted with a mixture of nitric, hydrochloric and sulphuric acids and determined by cold vapour atomic absorption. Sediments were oven-dried at 60'C, before being similarly extracted. Results showed that mercury in sediments at all sites in Trinidad and Venezuela exceeded NOAA and Canadian sediment quality guidelines, while cadmium, copper, nickel, lead and zinc also exceeded these guidelines at several sites. Heavy metal levels in oysters and green mussels varied widely with location. However, oysters from the Gulf of Paria contained significantly higher mean levels of cadmium, copper, nickel and zinc than those from the north coast of Venezuela, but this difference was not apparent in mussels. Cadmium, mercury and zinc in sediments were significantly correlated with those of mussels, but not of oysters, in which copper and zinc at several sites in the Gulf of Paria exceeded local maximum permissible levels (Cu = 20 microg g(-1) wet wt; Zn = 50 microg g(-1) wet wt) for human consumption. These findings indicate that while mussels may be better biological indicators of heavy metal pollution in sediments than oysters, the latter may provide copper and zinc contamination. Further research is needed to determine the most appropriate biological indicators of heavy metal and other pollutants in the local marine environment and to develop protocols for their use. PMID- 17465144 TI - Effects of herbicide on the kidneys of two Venezuelan cultured fish: Caquetaia kraussii and Colossoma macropomum (Pisces: Ciclidae and Characeae). AB - The use of chemical pesticides and herbicides has increased environmental pollution and affected ichthyofauna in the watersheds where they are used. We studied the effect of an herbicide, triazine, on the kidneys of two species (Caquetaia kraussii and Colossomna macropomum) widely found in Caribbean and South American rivers. In Venezuela, these species are abundant and have a high aquaculture potential because they may be cultured and reproduced in captivity. Four kidney samples from juveniles of each species exposed to the herbicide were examined by Transmission Electron Microscopy. Kidney tubule alterations included loss of plasmalemma and cell interdigitations, misshaped mitochondria, decrease in rough endoplasmic reticulum and free polysomes, and the presence of autophagic vacuoles and primary lysosomes. These alterations at the cellular level may explain fish behaviour in terms of kidney tubule pathology, and relative amounts and conditions of organelles within affected cells. PMID- 17465145 TI - The occurrence of black corals in Jamaican reef environments, with special reference to Stichopathes lutkeni (Antipatharia: Antipathidae). AB - The purpose of this study was to record the species of Antipatharia on Jamaican reefs and to carry out limited studies on densities and sizes of the common species. In addition, a cliff face created by dredging in 2002 provided the opportunity to study growth of newly settled colonies. Observations since 1998 and measurements since 2001 were made using SCUBA at depths down to 35 m. Seven species of Antipatharia were observed on steep coral reef escarpments below 25 m depth. The commonest species was the unbranched "wire coral" Stichopathes lutkeni. Other common species included the fan-shaped black corals Antipathes atlantica and A. gracilis. Frequently encountered species included commercially important A. caribbeana and a species with an unusual, scrambling growth form, A. rubusiformis. The other major commercial species in the Caribbean, Plumapathes pennacea, and a cave-dwelling species, A. umbratica, were rarely observed. Greatest black coral abundance occurred on steep slopes of hard substrata in low light intensity but exposed to the long-shore current. Combined densities of the commoner Antipatharia at 30 m deep at Rio Bueno on the north coast, ranged from 0.1 to 2.5 m(-2) (eleven 10 m x 1 m belt transects, 1-25 colonies per transect, 68 colonies in total). Forty-six of the 68 colonies were S. lutkeni, while nearby at Discovery Bay at 30-35 m, 55 out of 59 colonies were S. lutkeni. There was a significant difference between the mean length of colonies in these two populations of S. lutkeni (100 cm and 80 cm, respectively), probably relating to habitat. A third population of S. lukeni growing at 15-20 m deep on the recently dredged cliff had a much smaller mean length of 36.6 cm (n= 27). The largest individual measured 83 cm long, indicating a minimum growth rate of the unbranched corallum of 2.1 mm per day. PMID- 17465146 TI - [Fenology and life cycle of Gracilariopsis tenuifions (Gracilariaceae) at Sucre, Venezuela]. AB - The aim of this work was to determine the frequency of different reproductive phases and to induce the germination of spores from tetrasporic and cystocarpic Gracilariopsis tenuifrons from Chacopata and La Pena, Venezuela, under controlled laboratory conditions (temperature 22 +/- 1 degree C, 12L:12D photoperiod, salinity of 36 +/-1 PSU and irradiance of 269 microE m(-2) s(-1)). Tetrasporic individuals dominated numerically over gametophitic individuals. The proportion of vegetative algae was very low. Only cystocarpic algae were collected and the spermatangia were absent. Sporulation, germination and formation of algae suggest that they present a Polysiphonia-type life cycle. Algae with reproductive structures were not obtained in the laboratory, PMID- 17465147 TI - Coral recruitment patterns in the Florida Keys. AB - This study examines scleractinian zooxanthellate coral recruitment patterns in the Florida Keys to determine if differences in density or community composition exist between regions. From July to September 2002, nine patch reefs, three in each of the upper, middle and lower Keys, were surveyed for coral recruits (colonies <5 cm in diameter) using randomly placed quadrats and transects. Coral recruits were enumerated, measured, and identified to genus. Fourteen genera of corals were observed across all sites and ranged from five to 13 per site. Densities ranged from 6.29 +/- 1.92 (mean +/- SE) to 39.08 +/- 4.53 recruits m( 2), and there were significant site and regional differences in recruit densities. The density of recruits in the upper Keys was significantly lower than in the middle and lower Keys. In addition, the upper Keys were less diverse and had a different recruit size-frequency distribution. The majority of recruits were non-massive scleractinian species that contribute relatively little to overall reef-building processes, a finding that is similar to previous studies. Fewer recruits of massive species were found in the upper Keys compared to the middle and lower Keys. The recruitment patterns of the reefs in the upper Keys could potentially hinder their ability to recover from stress and disturbances. PMID- 17465148 TI - Patterns of sexual recruitment of acroporid coral populations on the west fore reef at Discovery Bay, Jamaica. AB - Coral recruitment was examined on terracotta tiles deployed for four six-month periods between March 2001 and April 2003 on the West Fore Reef at Discovery Bay, Jamaica. During each sampling period, four tiles were deployed on each of two arrays at six depths ranging from 3 m to 33 m. Only three Acropora spat recruited to the tiles over the sampling period. The Acropora spat recruited during only one of the four six-month sampling periods and at only one depth, 3m. That represents a density of 8 spat m(-2) at 3 m depth for one six-month sampling period. Acropora recruitment represented <1 % of the total spat recruiting to the tiles deployed at 3 m during the four sampling periods. Density of acroporids on the West Fore Reef is low. Only one Acropora colony (an A. palmata) was recorded during Point-Quarter surveys of coral cover and density at depths of 3 m, 9 m, 14 m and 19 m. Considering the paucity of acroporid colonies and the infrequent settlement of acroporid spat on the West Fore Reef, it is unlikely that the historic abundance of A. palmata and A. cervicornis will return soon. PMID- 17465149 TI - Potential sources of bacteriological pollution for two bays with marinas in Trinidad. AB - Welcome Bay and Chaguaramas Bay in the northwest peninsula of Trinidad contain large marinas and smaller sections of bathing beaches. Bacteriological surveys were conducted at both bays to assess water quality and to determine potential sources of pollution. These surveys were conducted during the wet season of 1996 and the dry season of 1997. Eleven sample stations were established at Welcome Bay and 12 at Chaguaramas Bay. Freshwater samples were collected from rivers and drains within the survey area. Marine water samples were collected from marinas, bathing beaches and inshore and outer areas at both bays. Five water samples were collected from each sampling station during the wet season of 1996 and six during the dry season of 1997. The membrane filter technique was used to determine faecal coliform and Escherichia coli levels in all samples. There was a seasonal effect on water quality, with significantly higher faecal coliform levels in the wet season, when water quality was not in compliance with international standards. This represents a potential health risk in bathing areas. Water quality was better at the outer area of both bays. Water quality at the inner bay areas was most likely adversely affected by land-based sources of pollution identified in this study. These sources include three drains and two rivers, which discharged into the bays. Yachts were apparently not a source of sewage pollution: there was no significant relationship between yacht number and faecal coliform levels. PMID- 17465150 TI - Transplantation of benthic species to mitigate impacts of coastal development in Jamaica. AB - Maintaining regional competitiveness and economic viability for Port Bustamante - Kingston Harbour, Jamaica, required improved accessibility to "Post Panamax" (too large to pass through the Panama Canal) container vessels. Removal of the northern portion of the shallow coral reef at Rackham's Cay, which was partially obstructing the western end of the east ship channel, was proposed. This aesthetically valuable reef was used by local fishermen and comprises part of the declared Palisadoes - Port Royal Protected Area. The proposal to transplant certain of the benthic species was advanced to mitigate loss of viable reef components. Between December 2001 and February 2002, sixty thousand items, consisting of reef building massive and branching corals; gorgonians; urchins (Diadema and Tripneustes spp.) and Thalassia meristems were relocated. During dredging, sedimentation rates from suspended solids in the water column were 0.003 g/cm(2)/day at the control site and 0.008 g/cm(2)/day at the dredge site. Coral cover in the relocation area increased from 15% to 20% while bare substrate decreased from 27% to 21%. This paper documents the mitigation required; some factors controlling the ecology of Rackham's Cay reef, the methodology of the relocation process; and the level of post-dredging survivorship of relocated corals. Political and economic realities of some proposed developments often override ecological considerations. Transplantation of important marine benthic species although time consuming, technically challenging, and expensive, may be one way for developers and ecologists to achieve sometimes disparate goals. This project cost US$1.7 million. The "items" moved were neither unique nor endemic and remain vulnerable to natural and anthropogenic impacts. This project increased public awareness and interest regarding the ecological and economic importance of reef ecosystems. It is anticipated that future coastal and inland developments will benefit from the lessons taught by these mitigative interventions. PMID- 17465151 TI - Aerial and tidal transport of mosquito control pesticides into the Florida Keys National Marine Sanctuary. AB - This project was undertaken as the initial monitoring program to determine if mosquito adulticides applied along the Florida Keys cause adverse ecological effects in the Florida Keys National Marine Sanctuary (FKNMS). The study monitored the distribution and persistence of two mosquito adulticides, permethrin and dibrom (naled), during three separate routine applications by the Florida Keys Mosquito Control District. The approach was to determine if toxic concentrations of the pesticides entered the FKNMS by aerial drift or tidal transport. The amount of pesticide entering the FKNMS by way of aerial drift was monitored by collection on glass fiber filter pads, set on floats in a grid pattern on either side of the FKNMS. Permethrin was recovered from filter pads on the leeward side for each of the three applications, ranging from 0.5 to 50.1 microg/m(2) throughout the study. Tidal current transport was monitored by collection of surface and subsurface water samples at each grid site. Tidal transport of naled and dichlorvos (naled degradation product) was apparent in the adjacent waters of the FKNMS. These compounds were detected in subsurface, offshore water at 0.1 to 0.6 microg/1, 14 hr after application. Permethrin was not detected in offshore water samples; however, concentrations ranging from 5.1 to 9.4 microg/l were found in surface water from the canal system adjacent to the application route. Comparison of the observed environmental concentrations with toxicity data (permethrin LC-50, 96 hr for Mysidopsis bahia = 0.02 microg/l) indicated a potential hazard to marine invertebrates in the canals with possible tidal transport to other areas. PMID- 17465152 TI - The importance of the marine ornamental reef fish trade in the wider Caribbean. AB - The marine ornamental fish trade began in the 1930s in Sri Lanka, spread to Hawaii and the Philippines in the 1950s, and expanded to a multi-million dollar industry in the 1970s with fisheries established throughout the tropical Pacific, Indian and Atlantic Oceans. Currently, 45 countries supply global markets an estimated 14-30 million fish annually, with an import value of US$28-44 million. The largest suppliers are Indonesia and the Philippines, followed by Brazil, Maldives, Vietnam, Sri Lanka and Hawaii. In the tropical Western Atlantic, 16 countries have export fisheries, including the U.S. (Florida and Puerto Rico). The U.S. is the world's largest buyer, followed by the European Union and Japan. The global trade consists of over 1400 species of reef fishes, of which only about 25 are captive bred on a commercial scale. Damselfish, anemonefish, and angelfish constitute over 50% of the global volume; butterflyfish, wrasses, blennies, gobies, triggerfish, filcfish, hawkfishes, groupers and basselets account for 31% of the trade, and the remaining 16% is represented by 33 families. The most important fishes from the Caribbean are angelfish (six species), seahorses (two species), royal gramma, jawfish, queen triggerfish, redlip blenny, puddingwife, bluehead wrasse, and blue chromis. The Caribbean currently supplies a small percentage of the global trade in marine ornamental species, but ornamental fisheries in this region represent important emerging industries. It is critical that effective ornamental fishery management plans and regulations are developed and enforced, and fishery-dependent and fishery independent data are collected and utilized in decision making processes to ensure sustainable ornamental fisheries throughout the region. PMID- 17465153 TI - Development of management policy for the marine ornamental fish and invertebrate fishery in puerto rico: a case study. AB - In recent years the collection of tropical marine organisms for the aquarium trade has become perceived as an activity with an unsustainable history as well as obvious potential for rehabilitation through resource-based fisheries management and consumer-oriented product certification. In the case of Puerto Rico, collection of ornamentals has existed for decades, though unregulated due to a weak fisheries law dating from the 1930's. The new Fisheries Law 278 of 1998 enabled new regulatory approaches for marine ornamentals, which were met with serious challenges rooted in (1) an information gap concerning the fishery regarding participant numbers, collection methods and export volumes, and (2) the absence of consultation of fishers by agency regulators. The information gap led to worst-case assumptions of impact by regulators, and a closure of the fishery, which set the stage for threatening personal confrontations and lawsuits, the latter leading to de facto resource management by judicial order. To redress these issues and move management back into the arena of science and public policy, regulators have initiated a three-phase program: (1) characterize fisher numbers, methods and exports, (2) describe populations and biology of commercial species, and (3) propose appropriate fisheries management approaches. This paper describes only the first phase of this program. PMID- 17465154 TI - Organization and operation of the marine ornamental fish and invertebrate export fishery in Puerto Rico. AB - This fishery was examined utilizing public records, stakeholder interviews, and operational site visits to describe the fishery for the Puerto Rico Coral Reef Advisory Committee as a first step toward development of policies for the effective management of these natural resources. The fishery is not large, including fewer than 20 licensed fishers operating primarily on the west end of the island. Only three operators export product, with the remaining fishers providing specimens to the exporters based upon customer orders. Most collection of coral reef species occurs over hard rubble zones mixed with relic reef structures and rock, or on the sides and frontal areas of active reefs. Other species are collected from among mangrove prop root zones, tidal flats, and seagrass beds. Collections are made using simple barrier and dip nets for fish and motile invertebrates such as shrimp. Invertebrates such as crabs, starfish, and sea cucumbers are commonly collected by overturning small rocks, gathering the specimens, and then replacing the rocks in their original positions. Specimens are carried to the boat and transferred to individual cup holders to maximize survival. Although statements concerning former use of chemicals to assist capture were noted, no evidence of current chemical use was observed. Specimens are held in re-circulating seawater systems onshore until collections are aggregated and shipped. The fishery strives to operate with mortality of<1%, as mortalities of>3% are described as unacceptable to customers. More than 100 fish species are collected in this fishery, but the top ten species account for >70% of the total numbers and >60% of the total value of the fishery, with a single species, Gramma loreto (Royal Gramma), comprising >40% of the numbers. More than 100 species of invertebrates are collected, but this fishery is also dominated by a handful of species, including anemones, hermit crabs, turbo snails, serpent starfish, and feather duster polychaetes. PMID- 17465155 TI - Linking larval history to juvenile demography in the bicolor damselfish Stegastes partitus (Perciformes: Pomacentridae). AB - Otolith-based reconstructions of daily larval growth increments were used to examine the effect of variation in larval growth on size and age at settlement and post-settlement growth, survival and habitat preferences of juvenile bicolor damselfish (Stegastes partitus Poey). During August 1992 and 1994, newly settled S. partitus were collected from Montastraea coral heads and Porites rubble piles in Tague Bay, St. Croix, U.S. Virgin Islands (17 degrees 45'N, 64 degres 42' W). Daily lapillar otolith increments from each fish were counted and measured with Optimas image analysis software. S. partitus pelagic larval duration was 23.7 d in 1992 (n = 70) and 24.6 d in 1994 (n = 38) and larval age at settlement averaged 13.0 mm total length both years. Analysis of daily otolith increments demonstrated that variation in larval growth rates and size and age at settlement had no detectable effect on post-settlement survivorship but that larger larvae showed a preference for Montastraea coral at settlement. Late larval and early juvenile growth rates showed a significant positive relationship indicating that growth patterns established during the planktonic stage can span metamorphosis and continue into the benthic juvenile phase. Larval growth rates during the first two weeks post-hatching also had a strong effect on age to developmental competence (ability to undergo metamorphosis) in both 1992 and 1994 with the fastest growing larvae being 8 d younger and 0.8 mm smaller at settlement than the slowest growing larvae. These differential growth rates in early stage larvae established trajectories toward larval developmental competence and may prove important in biogeographical studies of larval dispersal. PMID- 17465156 TI - Determination of the distribution of shallow-water seagrass and drift algae communities with acoustic seafloor discrimination. AB - The spatial distribution of seagrass and algae communities can be difficult to determine in large, shallow lagoon systems where high turbidity prevents the use of optical methods like aerial photography or satellite imagery. Further complications can arise when algae are not permanently attached to the substratum and drift with tides and currents. A study using acoustic seafloor discrimination was conducted in the Indian River Lagoon (Florida, USA) to determine the extent of drift algae and seagrass. Acoustic surveys using the QTC View V system based on 50 and 200 kHz transducers were conducted near Sebastian Inlet. Results indicate that areas of seagrass can be identified, and are mixed with a high abundance of drift algae. Nearest-neighbor extrapolation was used to fill in spaces between survey lines and thus obtain spatially cohesive maps. These maps were then ground-truthed using data from towed video and compared using confusion matrices, The maps showed a high level of agreement (60%) with the actual distribution of algae, however some confusion existed between bare sand and algae as well as seagrass. PMID- 17465157 TI - Assessing the accuracy of acoustic seabed classification for mapping coral reef environments in South Florida (Broward County, USA). AB - The Atlantic coast of Broward County, Florida (USA) is paralleled by a series of progressively deeper, shore-parallel coral reef communities. Two of these reef systems are drowned early Holocene coral reefs of 5 ky and 7 ky uncorrected radiocarbon age. Despite the case of access to these reefs, and their major contribution to the local economy, accurate benthic habitat maps of the area are not available. Ecological studies have shown that different benthic communities (i.e. communities composed of different biological taxa) exist along several spatial gradients on all reefs. Since these studies are limited by time and spatial extent, acoustic surveys with the QTCView V bottom classification system based on a 50 kHz transducer were used as an alternative method of producing habitat maps. From the acoustic data of a 3.1 km(2) survey area, spatial prediction maps were created for the area. These were compared with habitat maps interpreted from in situ data and Laser Airborne Depth Sounder (LADS) bathymetry, in order to ground-truth the remotely sensed data. An error matrix was used to quantitatively determine the accuracy of the acoustically derived spatial prediction model against the maps derived from the in situ and LADS data sets. Confusion analysis of 100 random points showed that the system was able to distinguish areas of reef from areas of rubble and sand with an overall accuracy of 61%. When asked to detect more subtle spatial differences, for example, those between distinct reef communities, the classification was only about 40% accurate. We discuss to what degree a synthesis of acoustic and in situ techniques can provide accurate habitat maps in coral reef environments, and conclude that acoustic methods were able to reflect the spatial extent and composition of at least three different biological communities. PMID- 17465158 TI - Summit-to-sea mapping and change detection using satellite imagery: tools for conservation and management of coral reefs. AB - Continuous summit-to-sea maps showing both land features and shallow-water coral reefs have been completed in Puerto Rico and the U.S. Virgin Islands, using circa 2000 Landsat 7 Enhanced Thematic Mapper (ETM+) Imagery. Continuous land/sea terrain was mapped by merging Digital Elevation Models (DEM) with satellite derived bathymetry. Benthic habitat characterizations were created by unsupervised classifications of Landsat imagery clustered using field data, and produced maps with an estimated overall accuracy of>75% (Tau coefficient >0.65). These were merged with Geocover-LC (land use/land cover) data to create continuous land/ sea cover maps. Image pairs from different dates were analyzed using Principle Components Analysis (PCA) in order to detect areas of change in the marine environment over two different time intervals: 2000 to 2001, and 1991 to 2003. This activity demonstrates the capabilities of Landsat imagery to produce continuous summit-to-sea maps, as well as detect certain changes in the shallow-water marine environment, providing a valuable tool for efficient coastal zone monitoring and effective management and conservation. PMID- 17465159 TI - A geographic information system (GIS) analysis for trace metal assessment of sediments in the Gulf of Paria, Trinidad. AB - The Gulf of Paria is a semi-enclosed shallow basin with increasing coastal development activities along Trinidad's west coast. Sediments present a host for trace metal pollutants from overlying waters, therefore determination of their content is critical in evaluating and detecting sources of marine pollution. This paper presents a Geographic Information System (GIS) analysis of geochemical assessment for trace metals in coastal sediments of the Gulf of Paria. This GIS approach facilitates interpretation of the spatial relationships among key environmental processes. The GIS development involves the integration of spatial and attribute data pertaining to bathymetry, current systems, topography, rivers, land use/land cover and coastal sediments. It employs spatial interpolation and retrieval operations to analyze the total trace metal concentrations of aluminum, copper and lead in the sediments and the clay-enriched sediments, to determine whether they are related to sediment type or are affected by the discharge from anthropogenic sources. Spatial distribution modeling of element concentrations are produced to indicate contamination plumes from possible anthropogenic sources such as rivers entering the Gulf of Paria, and to reveal potential hot spots and dispersion patterns. A direct spatial correlation between clay-enriched sediments and high concentrations of aluminum and lead is detected, however regions of high concentrations of copper and lead indicate a relationship to anthropogenic sources. The effectiveness of GIS for visualization, spatial query and overlay of geochemical analysis is demonstrated. PMID- 17465160 TI - Biomass, litterfall and decomposition rates for the fringed rhizophora mangle forest lining the Bon Accord Lagoon, Tobago. AB - The mangrove forest that fringes the Bon Accord Lagoon measures 0.8 km(2) and is dominated by red mangrove (Rhizophora mangle). This forest forms the landward boundary of the Buccoo Reef Marine Park in Southwest Tobago, and is part of a mangrove-seagrass-coral reef continuum. Biomass and productivity, as indicated by litterfall rates, were measured in seven 0.01 ha monospecific plots from February 1998 to February 1999, and decomposition rates were determined. Red mangrove above-ground biomass ranged between 2.0 and 25.9 kg (dry wt.) m(-2). Mean biomass was 14.1+/-8.1 kg (dry wt.) m(-2) yielding a standing crop of 11 318+/-6 488 t. Litterfall rate varied spatially and seasonally. It peaked from May to August (4.2-4.3 g dry wt. m(-2) d(-1)) and was lowest from October to December (2.3-2.8 g dry wt. m(-2) d(-1)). Mean annual litterfall rate was 3.4+/-0.9 g dry wt. m(-2) d(-1). Leaf degradation rates ranged from 0.3% loss d(-1) in the upper intertidal zone to 1% loss d(-1) at a lower intertidal site flooded by sewage effluent. Mean degradation rate was 0.4+/-1% loss d(-1) . The swamp produces 2.8 t dry wt. of litterfall and 12 kg dry wt. of decomposed leaf material daily. Biomass and litterfall rates in Bon Accord Lagoon were compared to five similar sites that also participate in the Caribbean Coastal Marine Productivity Programme (CARICOMP). The Bon Accord Lagoon mangrove swamp is a highly productive fringed forest that contributes to the overall productivity of the mangrove-seagrass-reef complex. PMID- 17465161 TI - The structure and productivity of the Thalassia testudinum community in Bon Accord Lagoon, Tobago. AB - The Thalassia testudinum dominated seagrass community in the Buccoo Reef/ Bon Accord Lagoon Marine Park, measures 0.5 km(2) and is part of a contiguous coral reef, seagrass bed and mangrove swamp system in southwest Tobago. 7 testudinum coverage, productivity and percent turnover rates were measured from February 1998 to February 1999 at four sample locations, while total T. testudinum biomass was measured at two locations in the lagoon from 1992-2002. Productivity and turnover rates varied spatially and seasonally. They were higher in the back-reef area than in the mangrove-fringed lagoon, and were lowest at locations near to a sewage outfall. 7 testudinum coverage ranged from 6.6% in the lagoon to 68.5% in the back-reef area while productivity ranged from 3.9 to 4.9 g dry wt m(-2) d(-1) . Productivity and percentage turnover rates were higher in the dry season (January-June) than in the wet season (July-December). Productivity ranged from 3.0 in the wet season to 5.0 g dry wt m2 d-' in the dry season while percentage turnover rates ranged from 4.2% to 5.6%. Total Thalassia biomass and productivity in Bon Accord Lagoon were compared to six similar sites in the Caribbean that also participate in the Caribbean Coastal Marine Productivity Program (CARICOMP). This seagrass community is being negatively impacted by nutrient-enriched conditions. PMID- 17465162 TI - Environmental education in Tobago's primary schools: a case study of coral reef education. AB - Environmental education is a relatively new area on the primary school curriculum of Trinidad and Tobago. Because of the close relationship between human activities and the degradation of the natural environment in Tobago, environmental education will become increasingly important to the preservation and conservation of the island's fragile natural resources. Current teaching methods rely heavily on text books and utilise a lecture style that does not promote student interaction. Unfortunately, these methods are not very conducive to environmental education. As such, this paper examines a pilot program in which staff from the Buccoo Reef Trust taught students from 15 primary schools about coral reefs using interactive tools and hands-on methods as described in People & Corals: an Education Package for Primary Schools (People & Corals). The pilot program ran over an eight week period with prepared lessons being taught every two weeks and student evaluations taking place once before the first lesson and once after the last lesson. The lessons were supplemented with a field trip to a coral reef ecosystem. Despite several challenges that were faced in the implementation process, the overall outcome of the pilot program was successful. Teachers and students reacted positively to the information that was being shared, thereby reinforcing the effectiveness of using a dynamic, active method of teaching to advance environmental education. PMID- 17465163 TI - Beach dynamics and nest distribution of the leatherback turtle (Dermochelys coriacea) at Grande Riviere Beach, Trinidad & Tobago. AB - Grande Riviere Beach in Trinidad and Tobago is an important nesting site in the Caribbean for the Critically Endangered leatherback sea turtle, Dermochelys coriacea. Community members were concerned that beach erosion and seasonal river flooding were destroying many of the nests deposited annually and thought that a hatchery was a possible solution. Over the 2001 turtle nesting season, the Institute of Marine Affairs (IMA) assessed the spatial and temporal distribution of nests using the Global Positioning System recorded to reference points, and beach dynamics using permanent bench mark profile stations, to determine areas of high risk and more stable areas for nesting. A total of 1449 leatherback nests were positioned. It was evident that at the start of the season in March, the majority of leatherback nests were deposited at the eastern section of the beach. After May, there was a continuing westward shift in nest distribution as the season progressed until August and beach erosion in the eastern section became predominant. The backshore remained relatively stable along the entire beach throughout the nesting season, and erosion was predominant in the foreshore at the eastern section of the beach, from the middle to the end of the season. Similar trends in accretion and erosion were observed in 2000. River flooding did not occur during the study period or in the previous year. With both high risk and more stable regions for turtle nesting available at Grande Riviere Beach, there was no compelling evidence to justify the need for a hatchery. PMID- 17465164 TI - The business role of healthcare quality professionals. PMID- 17465165 TI - Monitoring patient improvement and treatment outcomes in managed behavioral health. AB - This study examined the use of outcome reports sent to clinicians by a managed behavioral healthcare organization to monitor patient progress and its relation to treatment outcome. Results showed that clinicians who reported using outcome information had patients who also reported greater improvement at 6 months from baseline. Improvement per session was greatest among patients whose clinicians reported reading the outcome report and using outcome measures in their clinical practice. Using baseline and ongoing measures to assess patient improvement can provide clinicians with feedback during treatment, which may lead to better clinical outcomes and enable quality management systems in managed care to flag high-risk cases and identify failure of adequate improvement. PMID- 17465166 TI - Improving communication and documentation concerning preliminary and final radiology reports. AB - The communication and documentation of radiology preliminary and final interpretations for the identification of discrepancies failed to evolve with radiology's transition away from film. Processes should facilitate documentation and distribution of preliminary findings to multiple practitioners in real time and should provide timely communication and documentation of additional findings. The evolution of the electronic medical record and digital radiology images provided an opportunity to establish effective, safe, and efficient methods of communication and documentation. Froedtert Hospital (Milwaukee, WI) used the Six Sigma process to improve communication and documentation of radiologic findings. Root causes of problems were determined, hypotheses were tested, and new processes were piloted and implemented. Froedtert Hospital improved its process for documenting and communicating preliminary and final interpretations and identifying discrepancies. PMID- 17465167 TI - Trends in avoidable hospitalizations for children in Florida from 1992 to 2003. AB - This article examines trends in avoidable hospitalizations to determine whether improved access following post-1997 Balanced Budget Act expansions in public health insurance programs resulted in a decreased rate among affected populations, particularly children of ages 0-19 years. Florida inpatient discharge data sets from 1992 to 2003 were compiled in order to analyze trends using descriptive and linear regression methods. The rate of avoidable hospitalizations for children was subsequently compared to that of adults (ages 20-64 years) to assess relative changes. The rate declined significantly for black and other nonwhite children. Based on age, the declines were concentrated in the subgroups for ages 0-1 year, 5-9 years, and 10-14 years. In contrast, the rate for adults, with the exception of nonwhite and black populations, increased. The decline in the overall rate of avoidable hospitalizations for children indicates that increased expenditures related to the expansions were at least partially offset. However, the trend could not be directly attributed to the Balanced Budget Act. PMID- 17465168 TI - Glycemic control and use of the insulin sliding scale in hospitalized patients with diabetes. AB - This study assessed the extent of use of the subcutaneous insulin sliding scale (ISS) with hospitalized medical patients and examined the association between ISS use and glucose control. Despite some concerns about efficacy and suitability, the ISS is often used in the hospital studied. Researchers reviewed records of patients with a secondary diagnosis of diabetes who were admitted to the medical teaching unit of a tertiary care hospital. On day 1, 45.2% of patients were on an ISS alone. Patients on an ISS were more likely to experience hyperglycemia compared with patients on scheduled regimens, but they also had fewer hypoglycemic episodes. PMID- 17465169 TI - A physician self-assessment tool for quality improvement. AB - A physician self-assessment (PSA) tool was evaluated as an alternative to full chart reviews to assess guideline adherence. PSAs, consisting of brief self report questionnaires, were developed for asthma management and mammography screening and were mailed to participating physicians in a partially capitated Medicaid managed care insurance program. A random chart audit was conducted at each practice site, and results were compared to the PSA responses. An 85% concordance was found between the asthma PSA and chart data, and an 88% concordance was found between the mammography PSA and chart data. These findings suggest that the PSA, combined with random chart audits, can be a viable alternative to full chart reviews. PMID- 17465170 TI - William L. Roper on medication safety. PMID- 17465171 TI - Evidence-based monitoring for metabolic syndrome in clients with chronic schizophrenia. AB - Metabolic syndrome is a risk factor for cardiovascular disease. People with chronic schizophrenia are at risk for metabolic syndrome because of their diets, lifestyle, and (in some cases) their medication. The increased risk of metabolic syndrome has implications for the delivery of care to this population. This article provides an overview of metabolic syndrome in patients with schizophrenia and evidence-based criteria for monitoring this population. A recommendation is made to aggregate data collection in one place to facilitate follow-up. A sample form and letter are provided. PMID- 17465172 TI - Modern methods to treat superficial fungal disease. PMID- 17465173 TI - A second generation of active living research. PMID- 17465174 TI - Environment and active living: the roles of health risk and economic factors. AB - PURPOSE: This study examines the associations that a neighborhood ' physical and social environments have with transportation and recreation physical activities, with an emphasis on the roles of health risk and economic factors. DESIGN: It is a cross-sectional study with a hypothesis-testing approach. Setting. The study was conducted within the city of Seattle, Washington. Subjects. The subjects included 438 able-bodied, randomly selected adults. MEASURES: Physical activity and sociodemographic data came from a telephone survey (34 % response rate). Environmental variables were measured subjectively as people's perceptions and objectively using the Geographic Information System. Bivariate analyses and the Structural Equation Model were used to test the overall theoretic framework and the relationships among latent and observed variables. RESULTS: Lower-income populations lived in areas with more routine destinations and higher densities and were more active for transportation than higher-income populations. People with higher health risks were less active for both transportation and recreation purposes. The social environment-perception of people walking and biking in the neighborhood-was more strongly associated with recreational physical activities, while the physical environment was more strongly associated with transportation physical activities. CONCLUSION: Further investigation of different subpopulations and explicit distinction among different purposes of physical activities are needed in future research and interventions. This study is limited to urban areas and cross-sectional data. PMID- 17465175 TI - Urban form relationships with walk trip frequency and distance among youth. AB - PURPOSE: To assess the relationship among objectively measured urban form variables, age, and walking in youth. DESIGN: Cross-sectional analyses of travel diary data mapped against urban form characteristics within a 1-km buffer of participant's place of residence. Setting. Youth in the Atlanta, Georgia region with selection stratified by income, household size, and residential density. SUBJECTS: A total of 3161 5- to 20-year-olds who completed 2-day travel diaries. Diaries of those under 15 years were completed by a parent or legal guardian. MEASURES: Walking distances were calculated from a 2-day travel diary. Residential density, intersection density, land use mix, and commercial and recreation space were assessed within a 1-km network distance around residences. Analysis. Logistic regression analyses were performed for each urban form variable by age groups controlling for the demographic variables. All variables were then entered simultaneously into an analysis of the whole sample. RESULTS: All five urban form variables tested were related to walking. Recreation space was the only variables associated with walking across the four different age groups. All the urban form variables were related to walking in the 12 to 15 years age cohort. For this group, the odds of walking were 3. 7 times greater for those in highest- versus lowest-density tertile and 2.6 times greater for those with at least one commercial and 2.5 times greater for those with at least one recreational destination within 1 km from home. In the analysis of the full sample, number of cars, recreation space, and residential density were most strongly related to walking. CONCLUSIONS: Access to recreation or open space was the most important urban form variable related to walking for all age groups. Children aged 12 to 15 years old may be particularly influenced by urban form. PMID- 17465176 TI - Determinants of activity-friendly neighborhoods for children: results from the SPACE study. AB - PURPOSE: To examine the association between children's physical activity and factors of the built environment. DESIGN: Cross-sectional study. Setting. Ten neighborhoods in six cities in the Netherlands. SUBJECTS: Four hundred twenty-two children (age range, 6-11 years; 49% male). MEASURES: Physical activity diary, neighborhood observations, and anthropometric measures. ANALYSIS: Univariate and multivariate linear regression analyses. RESULTS: According to univariate analyses adjusted for age, sex, body mass index, and highest level of maternal education, physical activity (> or = 3 metabolic equivalents) was significantly (p < .05) associated with the proportion of green space, with the residential density, with the general impression of activity-friendliness of the neighborhood, and with the frequency of certain types of residences (e.g., terraced houses), sports fields, water, dog waste, heavy traffic, and safe walking and cycling conditions (e.g., cycle tracks and 30-km speed zones) in the neighborhood. According to adjusted multivariate analyses, physical activity was best predicted by the frequency of parallel parking spaces in the neighborhood and by the general impression of activity-friendliness of the neighborhood (I2 = 0. 193). CONCLUSIONS: Children's physical activity is associated with certain modifiable factors of the built environment. Longitudinal studies should examine whether there is a causal relationship. PMID- 17465177 TI - Green neighborhoods, food retail and childhood overweight: differences by population density. AB - PURPOSE: This study examines relationships between overweight in children and two environmentalfactors--amount of vegetation surrounding a child's place of residence and proximity of the child's residence to various types of food retail locations. We hypothesize that living in greener neighborhoods, farther from fast food restaurants, and closer to supermarkets would be associated with lower risk of overweight. DESIGN: Cross-sectional study. SETTING: Network of primary care pediatric clinics in Marion County, Indiana. SUBJECTS: We acquired data for 7334 subjects, ages 3 to 18 years, presenting for routine well-child care. MEASURES: Neighborhood vegetation and proximity to food retail were calculated using geographic information systems for each subject using circular and network buffers. Child weight status was defined using body mass index percentiles. Analysis. We used cumulative logit models to examine associations between an index of overweight, neighborhood vegetation, and food retail environment. RESULTS: After controlling for individual socio-demographics and neighborhood socioeconomic status, measures of vegetation and food retail significantly predicted overweight in children. Increased neighborhood vegetation was associated with decreased risk for overweight, but only for subjects residing in higher population density regions. Increased distance between a subject's residence and the nearest large brand name supermarkets was associated with increased risk of overweight, but only for subjects residing in lower population density regions. CONCLUSIONS: This research suggests that aspects of the built environment are determinants of child weight status, ostensibly by influencing physical activity and dietary behaviors. PMID- 17465178 TI - The urban built environment and obesity in New York City: a multilevel analysis. AB - PURPOSE: To examine whether urban form is associated with body size within a densely-settled city. DESIGN: Cross-sectional analysis using multilevel modeling to relate body mass index (BMI) to built environment resources. SETTING: Census tracts (n = 1989) within the five boroughs of New York City. SUBJECTS: Adult volunteers (n = 13,102) from the five boroughs of New York City recruited between January 2000 and December 2002. MEASURES: The dependent variable was objectively measured BMI. Independent variables included land use mix; bus and subway stop density; population density; and intersection density. Covariates included age, gender, race, education, and census tract-level poverty and race/ethnicity. ANALYSIS: Cross-sectional multilevel analyses. RESULTS: Mixed land use (Beta = .55, p < .01), density of bus stops (Beta = -.01, p < .01) and subway stops (Beta = -.06, p < .01), and population density (Beta = -.25, p < .001), but not intersection density (Beta = -. 002) were significantly inversely associated with BMI after adjustmentfor individual- and neighborhood-level sociodemographic characteristics. Comparing the 90th to the 10th percentile of each built environment variable, the predicted adjusted difference in BMI with increased mixed land use was -. 41 units, with bus stop density was -.33 units, with subway stop density was -.34 units, and with population density was -.86 units. CONCLUSION: BMI is associated with built environment characteristics in New York City. PMID- 17465179 TI - Trail characteristics as correlates of urban trail use. AB - PURPOSE: The purpose of this study was to identify the environmental correlates of urban trail use. DESIGN: Three multiuse urban trails at least 15 miles in length were selected. Trails were divided into one-half-mile segments (N = 102 total segments) and marked in the field. An audit of each trail was completed identifying built environmental features. A cross-sectional count of trail users was completed at each segment estimating the type of use, age, gender. SETTING: Data collection occurred on urban trails in Chicago, Dallas, and Los Angeles. SUBJECTS: Trained observers counted 17,338 users across the three trails. MEASURES: The SPACES for Trails instrument was used, and a trail count data collection sheet was developed. ANALYSIS: Univariate and multivariate Poisson regressions controlling for location of trail and density of the population within 1 mile of the trail. RESULTS: Positive associations with trail use were observed for mixed views (beta = .33, p < .0001), streetlights (beta = .30, p < .0001), good trail condition (beta = .28, p < .0001), and the presence of cafes (beta = .38, p < .0001) and other trailside facilities (beta =.08, p < .0001). Negative associations were observed for litter (beta = -.22, p < .0001), noise (beta = -.41, p < .0001), higher vegetation density (beta = -. 10, p < .001), drainage features (beta = -.67, p < .0003), natural areas adjacent to the trail (beta = -.39, p < .0001), and tunnel present (beta = -.20, p < .04). CONCLUSIONS: These correlates should be confirmed in other studies and if supported should be considered in the promotion and design of urban trails. PMID- 17465180 TI - Spatial measures associated with stair use. AB - PURPOSE: Although stair use in workplaces can provide an accessible means of integrating physical activity into work routines, there is little information available on how building design influences stair use. DESIGN: This cross sectional study assessed the relationship between stair use and the design and location of stairs. SETTING: Ten three- or four-story academic buildings on two university campuses. SAMPLE: The buildings contained a total of 38 stairs and 12 elevators. MEASURES: Stair use was measured using infrared monitors. Eighteen environmental variables that operationalized the appeal, convenience, comfort, legibility, and safety of stairs were measured. RESULTS: Regression analysis identified eight spatial variables associated with stair use: travel distances from stair to nearest entrance and the elevator, effective area or occupant load of each stair, accessibility of each stair, area of stair isovist (a graphic representation of the horizontal extent of a person's visual field from a specific point of reference within a building floor plan), number of turns required for travel from the stair to closest entrance, and the most integrated path (MIP). Three variables (effective area, area of stair isovist, and number of turns for travel from the MIP), explained 53 % of stair use in the 10 buildings. Most variables operationalizing the appeal, comfort, and safety of stairs were not statistically influential. CONCLUSIONS: This study suggests that the spatial qualities that optimize the convenience and legibility of stairs may have the most influence on stair use in buildings. PMID- 17465181 TI - The influence of the media environment on physical activity: looking for the big picture. AB - The Influence of the Media Environment on Physical Activity: Looking for the Big Picture Media consumption is expected to continue to increase well into the future. With this in mind the author suggests opportunities to influence the media environment for the purposes of promoting physical activity. A research agenda focused on media consumption, media content, commercial considerations, promoting physical activity through the media, partnerships with industry, positioning, dissemination of proven methods, and environmental change is proposed. PMID- 17465182 TI - The role of personal values, urban form, and auto availability in the analysis of walking for transportation. AB - PURPOSE: To examine the association of personal values, the built environment, and auto availability with walking for transportation. SETTING: Participants were drawn from 11 U.S. metropolitan areas with good transit services. SUBJECTS: 865 adults who had recently made or were contemplating making a residential move. MEASURES: Respondents reported if walking was their primary mode for nine trip purposes. "Personal values" reflected ratings of 15 variables assessing attitudes about urban and environmental attributes, with high reliability (ot = 0.85). Neighborhood form was indicated by a three-item scale. Three binary variables were created to reflect (1) personal values, (2) neighborhood form, and (3) auto availability. DESIGN: The association with walking was reported for each of the three variables, each combination of two variables, and the combination of three variables. An analysis of covariance was applied, and a hierarchic linear regression model was developed. RESULTS: All three variables were associated with walking, and all three variables interacted. The standardized coefficients were 0.23for neighborhood form, 0.21 for autos per person, and 0.18 for personal values. CONCLUSION: Positive attitudes about urban attributes, living in a supportive neighborhood, and low automobile availability significantly predicted more walking for transportation. A framework for further research is proposed in which a factor representing the role of the automobile is examined explicitly in addition to personal values and urban form. PMID- 17465183 TI - Using objective and subjective measures of neighborhood greenness and accessible destinations for understanding walking trips and BMI in Seattle, Washington. AB - PURPOSE: Examine the influence of destinations within walking distance of a residence and vegetation on walking trips and body mass index (BMI). DESIGN: Cross-sectional analysis of data from residences with varying accessibility and greenness. SETTING: Seattle, Washington. SUBJECTS: Stratified random sample of residents, stratified by accessibility and greenness. RESPONSE RATE: 17.5 %, 529 respondents. MEASURES: Accessibility and greenness were measured objectively by Geographic Information Systems (GIS) Network Analysis and normalized difference vegetation index (NDVI), respectively. Self-reported destinations, natural features, walking trips, BMI, and importance of destinations were measured through a postal survey. RESULTS: Objective accessibility were related to walking trips per month (r(2) = .110, p < .0001), as was subjective greenness (r(2) = .051, p < .0001), although objective measures of actual greenness were not. In areas with high accessibility, BMI was lower in areas that had high NDVI, or more greenness (r(2) = .129428, model p < .0001; t-test of interaction p = .0257). Low NDVI areas were associated with overestimation of the number of destinations within walking distance (F1, 499 = 11.009, p = .001). CONCLUSIONS: Objective and subjective measurements of accessibility and greenness led to an understanding of variation among walking trips and BMI in different neighborhoods. PMID- 17465184 TI - Crime prevention and active living. AB - This paper addresses the question of whether crime is a barrier to active living and if it is, what can be done about it? The authors introduce a theoretical model that addresses how crime might influence physical activity behavior. The core components of the model are: situational characteristics, crime and disorder, fear of crime or disorder, and physical activity. These variables are thought to be moderated through psychological, demographic, environmental and other factors. Research questions that derive from the model are featured. PMID- 17465185 TI - Schools as a community resource for physical activity: legal considerations for decision makers. AB - Public schools may offer community residents access to gymnasiums, playfields, etc. When school facilities are available and/or opened up for public use in this manner, what are the legal obligations and liability concerns that arise for the schools and the users? Joint- and shared-use facilities and lands have some legal protections, which this paper introduces and reviews. Legal cases (and precedent) are shared with readers. Within this context, a model depicting factors influencing decisions to allow public use of school facilities for recreation and physical activity is introduced. PMID- 17465186 TI - Promoting active community environments through land use and transportation planning. AB - PURPOSE: To examine the role of land use and transportation plans as policy instruments for promoting active community environments. DESIGN: Cross-sectional analysis using multilevel models to examine whether active community environment scores were associated with leisure and transportation-related physical activity (PA) and whether associations varied by household income. SETTING: 67 North Carolina counties SUBJECTS: Adults (n = 6694) from pooled 2000 and 2002 North Carolina Behavioral Risk Factor Surveillance System (BRFSS) surveys. MEASURES: Active community environment scores, derived from a 2003 survey of planning directors, representing the presence of nonmotorized transportation improvements, mixed land use classification, and comprehensiveness of implementation tools. Dependent variables were self-reported PA measures from the BRFSS. Sociodemographic variables were derived from the 2000 U.S. Census of Population. RESULTS: After adjustment for sociodemographic factors, more favorable active community environment scores were significantly associated with leisure PA (p = .001), transportation PA (p < .01), bicycling (p < .05), walking 150 minutes/week (p < .001), and meeting PA recommendations (p < .0001). In stratified analyses, lower-income individuals (<$25, 000) living in high scoring counties were three times more likely to participate in transportation PA compared with those living in low scoring counties (95% confidence interval, 1.4, 7.3). CONCLUSIONS: This study identifies previously unexamined policy and institutional correlates of PA related to land use and transportation planning. Plans may provide a means to incorporate community support for active living into public policy. PMID- 17465187 TI - [Obesity and its implication in gastroenterological diseases]. PMID- 17465188 TI - [Management of upper gastrointestinal hemorrhage]. PMID- 17465189 TI - [Gastroenterology in the elderly]. PMID- 17465190 TI - [Hepatic amebiasis]. PMID- 17465191 TI - [Hepatitis A and B vaccines]. PMID- 17465192 TI - Gene expression analysis of the mechanisms whereby black cohosh inhibits human breast cancer cell growth. AB - BACKGROUND: Previous studies indicate that specific extracts and the pure triterpene glycoside actein obtained from black cohosh inhibit growth of human breast cancer cells. Our aim is to identify alterations in gene expression induced by treatment with a methanolic extract (MeOH) of black cohosh. MATERIALS AND METHODS: We treated MDA-MB-453 human breast cancer cells with the MeOH extract at 40 microg/ml and collected RNA at 6 and 24 h; we confirmed the microarray results with real-time RT-PCR for 18 genes. RESULTS: At 6 h after treatment there was significant increase in expression of ER stress (GRP78), apoptotic (GDF15), lipid biosynthetic (INSIG1 and HSD17B7) and Phase 1 (CYP1A1) genes and, at 24 h, decrease in expression of cell cycle (HELLS and PLK4) genes. CONCLUSION: Since the MeOH extract activated genes that enhance apoptosis and repressed cell cycle genes, it may be useful in the prevention and therapy of breast cancer. PMID- 17465193 TI - The camptothecin derivative CPT-11 inhibits angiogenesis in a dual-color imageable orthotopic metastatic nude mouse model of human colon cancer. AB - Recent studies have shown the expression of a stem cell marker protein, nestin, in nascent blood vessels in nestin-driven green fluorescent protein (ND-GFP) transgenic nude mice. In the present study, we visualized tumor angiogenesis and evaluated the antiangiogenic efficacy of CPT-11 in ND-GFP nude mice using dual color fluorescence imaging. We orthotopically implanted ND-GFP nude mice with the human cancer cell line HCT-116 expressing red fluorescent protein (RFP). The mice were treated with CPT-11 at 40 mg/kg on days 7, 10, 14. Tumor angiogenesis was imaged and visualized by dual-color fluorescence imaging on day 17, three days after the last CPT-11 treatment. Tumor volume and the mean nascent blood vessel density were determined and compared to the control mice. The growing tumor had high expressions of nestin in the nascent blood vessels. The nascent blood vessels showed co-localization of the endothelial-cell-specific marker CD-31 under immunohistochemical staining. The nascent blood vessels were highly visible and their density was determined. ND-GFP nude mice that were administered CPT-11 showed significant reduction in the mean nascent blood vessel density and tumor volume. The dual-color model of ND-GFP transgenic nude mice orthotopically implanted with HCT-116 expressing RFP proved to be effective in visualizing and quantitating tumor growth and tumor angiogenesis. The results showed that CPT-11 is an effective inhibitor of angiogenesis and provided strong implications for wider clinical application of CPT-11 for colon cancer. PMID- 17465194 TI - Cyclooxygenase-2 directly induces MCF-7 breast tumor cells to develop into exponentially growing, highly angiogenic and regionally invasive human ductal carcinoma xenografts. AB - Based on our studies demonstrating first time evidence that the cyclooxygenase-2 (Cox-2) enzyme is abundant within invasive human breast tumors, we developed a clonally derived human breast tumor cell clone designated as MCF-7/Cox-2 Clone 10 by transfection of human Cox-2 cDNA into slow growing, Cox-2 null, non-metastatic MCF-7 human breast tumor cells. The present studies evaluated the biological characteristics of the MCF-7/Cox-2 Clone 10 human breast tumors compared to the characteristics of MCF-7/empty vector control tumors when grown in vivo following injection of 5x10(6) tumor cells into mammary fat pads of ovariectomized female Crl:Nu-Foxn1(nu) mice implanted with slow release 17-beta estradiol pellets. At 60 days after tumor cell injection, MCF-7/Cox-2 Clone 10 human breast tumors were 4-fold greater (p < 0.01) in volume than MCF-7/empty vector control tumors. MCF 7/Cox-2 Clone 10 human breast tumor xenografts were highly angiogenic based on histological observation of large-bore blood vessels, which was confirmed by immunohistochemical staining with anti-CD-31 antibody and quantitation of mean vessel density. MCF-7/Cox-2 Clone 10 human breast tumor cells were present within regional lymph nodes adjacent to mammary fat pads with their local invasion confirmed by Western blotting of Cox-2-protein. This unique Cox-2-dependent breast tumor model rapidly produces large, angiogenic, locally invasive human breast tumor xenografts in mammary fat pads of ovariectomized female Crl:Nu Foxn1(nu) mice at 42-60 days which recapitulate human breast ductal carcinomas. This unique model may be invaluable as a means to evaluate preclinical safety and efficacy of novel adjuvant therapies for women with metastastic breast cancer including prostanoid receptor antagonists, newly developed anti-angiogenic therapies, as well as other novel approaches for targeting and destruction of human breast tumors and their vasculature. PMID- 17465195 TI - Antiproliferating activity of the mitotic inhibitor pironetin against vindesine- and paclitaxel-resistant human small cell lung cancer H69 cells. AB - Pironetin, isolated from Streptomyces sp., is a potent inhibitor of microtubule assembly and the first compound identified that covalently binds to alpha-tubulin at Lys352. We examined whether pironetin is an effective agent against human small cell lung cancer H69 cells, including two cell lines resistant to the microtubule-targeted drugs vindesine (H69/VDS) and paclitaxel (H69/Txl) that interact with beta-tubulin. Pironetin was found to be effective against these resistant cells as well as their parental cells. In addition, pironetin inhibited the growth of human leukemic K562 multidrug-resistant cells (K562/ADM), which have mdr1 gene expression, as well as the parental K562 cells. In these cell lines, including the parental and resistant cells, pironetin caused complete mitotic arrest; in addition, apoptosis inductions by 30 and 100 nM pironetin were observed. In this study, the new mitotic inhibitor, pironetin, was found to be effective not only against human tumor cell lines resistant to microtubule targeted drugs, but also multidrug-resistant cells with mdr1 gene expression. These results suggest that pironetin is a useful agent for overcoming drug resistance in cancer chemotherapy. PMID- 17465196 TI - Sensitivity of bladder cancer cells to curcumin and its derivatives depends on the extracellular matrix. AB - Because the response of cancer cells to chemotherapeutic agents depends upon the supporting extracellular matrix (ECM), the response in vivo may not be reproduced in 2-dimensional cell culture. The dose-response to curcumin and two derivatives by bladder cancer cells grown on both normal (SISgel) and cancer-derived ECM (Matrigel) and on plastic were contrasted. Cells grown on Matrigel were resistant to curcumins, but cells growing on SISgel, which mimic cancer cells suppressed by normal ECM, were nearly as sensitive as cells grown on plastic. SV40-immortalized urothelial cells, which are models for premalignant cells, were the most sensitive, but even aggressive cell lines were nearly as sensitive when grown on SISgel as on plastic. Curcumin response depends highly on the supporting ECM, and cells grown on plastic poorly models cells growing on natural ECM. Curcumin could prove an effective chemopreventive for bladder cancer recurrence when administered intravesically post-therapy. PMID- 17465197 TI - Induction of differentiation of colon cancer cells by combined inhibition of kinases and histone deacetylase. AB - The MAP kinase pathway inhibitor U0126 in combination with butyrate promotes differentiation in some colon cancer cell lines. We examined several inhibitors of histone deacetylase (HDAC) in combination with U0126 and other protein kinase inhibitors to see if these effects are general properties of HDAC inhibitors or butyrate alone. Alkaline phosphatase and peptidase activities were examined as markers for cellular differentiation in the human colon cancer cell lines Caco-2 and HT29 and the minimally transformed NCM460. Several HDAC inhibitors caused greater increases of alkaline phosphatase in the cancer cells than in NCM460, in which butyrate was the only HDAC inhibitor that caused a consistent increase. Unlike the JNK and PKC inhibitors examined, the MEK 1/2 inhibitor U0126 induced alkaline phosphatase activity in Caco-2 as a single agent and caused additive effects with HDAC inhibitors. The PI-3 kinase inhibitor LY294002 had little effect alone but enhanced the response of most HDAC inhibitors as did the raf inhibitor GW5074. In addition to butyrate, several HDAC inhibitors can induce differentiation in colon cancer cells and the responses may be enhanced by U0126, GW5074 and LY294002. PMID- 17465198 TI - Chromosomal damage in two X-ray irradiated cell lines: influence of cell cycle stage and irradiation temperature. AB - The aim of this study was to investigate if irradiation with X-rays in different cell cycle phases resulted in a different response as measured with the micronucleus technique. In addition, the influence of irradiation temperature was investigated. MATERIALS AND METHODS: Cells from a non-transformed human fibroblast cell line, HS2429, and a human breast cancer cell line, MCF-7, were synchronized by thymidine block and irradiated at either 2 degrees C or 37 degrees C in the G1-, S- and G2/M-phases. After cytokinesis-block by cytochalasin B, the frequency of micronuclei was determined. RESULTS: Clear dose-response relationships were found. More micronuclei were detected in fibroblast cells irradiated in G1 and S than in G2/M, while the differences were not as prominent in MCF-7 cells. The irradiation temperature had no significant influence on the formation of micronuclei in either of the cell lines. CONCLUSION: The formation of micronuclei varies with the cell cycle stage at the time of irradiation. PMID- 17465199 TI - Targeted transarterial therapy of Vx-2 rabbit liver tumor with Yttrium-90 labeled ferromagnetic particles using an external magnetic field. AB - BACKGROUND: Our goal was to study the efficacy of liver cancer embolization with magnetically targeted Yttrium-90 labeled ferromagnetic particles and establish the biodistribution profile of these particles. MATERIALS AND METHODS: Of twenty rabbits, nine underwent transarterial radioembolization of implanted Vx-2 tumor with increasing 90Y-MTC doses, three were treated with carrier particles alone, four remained untreated and four were sacrificed early to document biodistribution. At various intervals, animals were sacrificed and biodistribution, liver cancer viability and toxicity were measured. RESULTS: There was a dose related degree of tumor necrosis, with greater than 90 Gy yielding 100% necrosis (baseline 50%). Blood radioactivity one hour post radioembolization was less than 0.0275 microCi/g. No hematological toxicity was observed. Except for the non-targeted right liver lobe, organ radioactivity levels were within tolerance levels. Significant left (targeted) hepatic lobe necrosis was seen in subjects receiving high doses. CONCLUSION: Hepatic arterial radioembolization with 9Y-MTC bolstered by external magnetic field has significant tumoricidal effect and a favorable biodistribution profile. PMID- 17465200 TI - LKB1 is crucial for TRAIL-mediated apoptosis induction in osteosarcoma. AB - BACKGROUND: Despite improvements in chemotherapy and surgery in the treatment of osteosarcoma, satisfactory results are still difficult to achieve. New therapeutic modalities need to be developed for the improvement of these treatments. TRAIL (TNF-related apoptosis inducing ligand) is known as a selective apoptosis inducer in most tumor cells, but not in normal cells. Therefore, TRAIL is a good candidate target for the treatment of tumors. However, sensitivity of osteosarcoma cells to TRAIL-induced apoptosis is lower than that of other types of tumor cells. Recently, DAP3 (death associated protein 3) was demonstrated to play a critical role in TRAIL-mediated apoptosis through activation of pro caspase-8. Here, we found that LKB1, a serine/threonine kinase, expressed in bone and soft tissue sarcoma cells, associated with DAP3. We also demonstrated that expression of DAP3 induced apoptosis in osteosarcoma cells. Furthermore, expression of LKB1 induced apoptosis and co-expression of LKB1 with DAP3 strongly induced apoptosis in osteosarcoma cells. In addition, expression of LKB1 kinase dead mutant, LKB1 (K78M), inhibited DAP3-induced apoptosis in these cells. These results suggest that LKB1 is critical for TRAIL-induced apoptosis induction, cooperating with DAP3 in osteosarcoma cells. It is predicted that LKB1 and DAP3 could be critical target molecules for the treatment of osteosarcomas. PMID- 17465201 TI - Antifolate pseudo-resistance due to elevated levels of thymidine and hypoxanthine in a commercial serum preparation. AB - BACKGROUND: Batch variability of sera used for cell culture is of considerable experimental concern. A novel fetal calf serum product, FCS Gold, was claimed to be the first defined fetal calf serum free of batch variation. MATERIALS AND METHODS: The efficacy of methotrexate (MTX) and LY231514 (multitargeted antifolate, MTA) in CCRF-CEM cells and KB cells was compared using media supplemented with FCS Gold or conventional fetal bovine serum. RESULTS: IC50 values from tests using conventional serum corresponded to published data. FCS Gold fully protected the cells from antifolate drug cytotoxicity. Dialysis of FCS Gold restored responsiveness to antifolate drugs. Elevated levels of hypoxanthine and thymidine were present in FCS Gold. They were approximately 10-fold greater than the concentrations required to overcome growth arrest mediated by 2 microM MTX. CONCLUSION: FCS Gold or identical products, e.g. FBS Gold, should not be used in studies on antifolate drug action. PMID- 17465202 TI - FEM analysis of RF breast ablation: multiprobe versus cool-tip electrode. AB - BACKGROUND: Radio-frequency ablation (RFA) has recently received much attention as an effective minimally invasive strategy for the local treatment of tumors. The purpose of this study was to evaluate the efficacy of single-needle cool-tip RF breast ablation in terms of temperature distribution and duration of the procedure as compared to multiprobe RF breast ablation. MATERIALS AND METHODS: Two different commercially available radiofrequency ablation needle electrodes were compared. Finite-element method (FEM) models were developed to simulate the thermoablation procedures. A series of ex vivo radiofrequency thermal lesions were induced to check the response of the FEM calculations. RESULTS: Data obtained from FEM models and from ex vivo procedures showed that cool-tip RF breast ablation assures better performances than multiprobe RF breast ablation in terms of temperature distribution and duration of the procedure. Histopathological analysis of the cool-tip RF thermoablated specimens showed successful induction of coagulation necrosis in the thermoablated specimens. CONCLUSION: Data obtained from FEM models and from ex vivo procedures suggest that the proposed cool-tip RF breast ablation may kill more tumor cells in vivo with a single application than the multiprobe RF breast ablation. PMID- 17465203 TI - Role and prognostic significance of proapoptotic proteins in Epstein-Barr virus infected gastric carcinomas. AB - BACKGROUND: The purposes of the present study were to evaluate the role and the prognostic values of proapoptotic proteins involved in the death receptors (Fas and TRAIL receptors) and mitochondrial pathways (Bax) in Epstein-Barr virus (EBV) infected gastric carcinomas. MATERIALS AND METHODS: Fifty-five EBV-infected gastric carcinomas were identified by in situ hybridization for EBV-encoded small RNAs. Immunohistochemistry was performed for Fas, Fas-ligand, FADD, TRAIL, DR4, DR5 and Bax. Apoptotic indices (AIs) were determined using TUNEL assay and assessed. RESULTS: No remarkable differences in protein expressions were observed between EBV-infected gastric carcinomas and conventional gastric carcinomas. Bax positivity tended to be associated with higher AI (p = 0.068), whereas Fas and FADD positivities were related to lower AI (p = 0.006 and 0.059, respectively). Proteins involved in TRAIL pathways showed no statistical significant relationship with AI. TNM stage and Fas and FADD expressions were related to overall survival (p < 0.05), but TNM stage was the only independent prognostic factor. CONCLUSION: Apoptosis in EBV-infected gastric carcinomas probably occurs via the mitochondrial pathway through Bax, rather than via the death receptor pathways. Fas and FADD expressions, and pathological tumor stage (TNM stage) may be the prognostic factors. PMID- 17465204 TI - Increased expression of ADAM 9 and ADAM 15 mRNA in pancreatic cancer. AB - BACKGROUND: A disintegrin and metalloproteases (ADAMs) comprise a multifunctional family of membrane-anchored proteins. ADAM 9 and ADAM 15 are involved in cell migration and invasion. Expression of ADAM 9 and ADAM 15 was reported to be altered in several types of cancer. MATERIALS AND METHODS: Quantitative real-time reverse transcription-polymerase chain reaction was performed to measure the expression of ADAM 9 mRNA in bulk pancreatic tissues. Results showed no significant difference in the expression of ADAM 9 mRNA between pancreatic cancer and non-neoplastic pancreas. Primary cultured pancreatic fibroblasts also expressed ADAM 9 mRNA. Therefore, a laser microdissection and pressure catapulting technique was employed to isolate cancer cells from tumor tissues. The expression of ADAM 9 and ADAM 15 mRNA was measured in microdissected samples (cancer cells, n = 11; normal epithelial cells, n = 13 for ADAM 9; cancer cells, n = 9; normal epithelial cells, n = 9 for ADAM 15). RESULTS: Pancreatic cancer cells expressed significantly higher levels of ADAM 9 and ADAM 15 mRNA than did normal pancreatic epithelial cells (p = 0.016 for ADAM 9; p = 0.004 for ADAM 15). CONCLUSION: ADAM 9 and ADAM 15 are involved in pancreatic cancer. Microdissection based analysis appears to be indispensable for the accurate analysis of the expression of certain ADAM family members in pancreatic cancer. PMID- 17465205 TI - Re-evaluation of cyclooxygenase-2-inhibiting activity of vanillin and guaiacol in macrophages stimulated with lipopolysaccharide. AB - Phytophenols such as para-substituted 2-methoxyphenols exhibit antioxidant and anti-inflammatory activities, however, their biological activities are concentration-dependent, possibly due to their dual property of being both antioxidant and prooxidant. Eugenol (2-allyl-2-methoxyphenol) and isoeugenol (4 propenyl-2-methoxyphenol) did not reveal cyclooxygenase-2 (COX-2)-inhibiting activity in macrophages stimulated with lipopolysaccharide (LPS). In contrast, vanillin (2-hydroxy-3-methoxybenzaldehyde) and guaiacol (2-methoxyphenol), especially the former, inhibited LPS-stimulated nuclear factor kappa B (NF kappaB) activation and cyclooxygenase (COX)-2 gene expression in cells of the RAW 264.7 murine macrophage cell line. Among the 2-methoxyphenols, vanillin demonstrated a potent anti-inflammatory activity. The phenolic O-H bond dissociation enthalpy (BDE) and molecular orbital energies (chemical hardness [eta], electronegativity [chi], and electrophilicity [omega]) were examined to clarify the mechanism responsible for inhibition of COX-2 expression. The BDE, chi, and omega values for vanillin were significantly higher than the corresponding values for the other 2-methoxyphenols. The anti-inflammatory activity of 2-methoxyphenols depended on the BDE and the phenol function was crucial for eliciting this activity. In addition, the anti-inflammatory activity depended on the chi and omega. These findings make vanillin attractive as a candidate therapeutic agent. PMID- 17465206 TI - Chemoembolization of lung metastases--pharmacokinetic behaviour of carboplatin in a rat model. AB - AIM: To improve tumor control in lung metastases using a novel method: unilateral chemoembolization of the lung by instillation of degradable starch microspheres (DSM) and cytotoxic agents via the pulmonal artery. MATERIALS AND METHODS: A rodent model of solitary metastasis (CC531 adenocarcinoma) was studied. The animals were randomized into three groups: the control group receiving carboplatin (45 mg/kg) intravenously, an isolated lung perfusion (ILP) group recieving buffered starch solution and carboplatin (15 mg/kg) and a third group receiving chemoembolization with carboplatin (15 mg/kg) and DSM (2 ml/kg). The total platinum concentration in serum, lung and lung tumor at defined times (15, 30, 60, 120 min) was measured using an inductively coupled plasma mass spectrometer (ICP-MS). RESULTS: The area under concentration (AUC) versus time curves showed a 7.9- to 42.6-fold higher concentration in the tumor tissue comparing the ILP and chemoembolization group to the control group (p < 0.01). In the comparison of the AUCs of ILP versus chemoembolization, the tumor tissue of the lung showed a 5.4-fold higher concentration in the chemoembolization group (p < 0.01). CONCLUSION: This is the first study to measure the concentration of carboplatin during chemoembolization of the lung. Compared to intravenous therapy, chemoembolization produced higher tumor tissue concentrations. Comparing chemoembolization to ILP, there was also an increase of carboplatin in the tumor tissue, without histological damage of the surrounding lung parenchyma. PMID- 17465207 TI - Functional alteration of myeloid dendritic cells through head and neck cancer. AB - BACKGROUND: In head and neck squamous cell carcinoma (HNSCC), the secretion of various immunosuppressive mediators contributes to large-scale effects on the immune functions. The influence of HNSCC on various cellular functions of human myeloid dendritic cells (MDC) was analyzed in this work. MATERIALS AND METHODS: MDCs were isolated from peripheral blood by 'magnetic bead separation'. Cellular functions were analyzed using flow cytometry, migration- and ELISPOT assays as well as cytokine detection assays. RESULTS: HNSCC massively affects MDCs to induce effective T-cell responses. Analysis of MDC migration and cytokine secretion revealed that HNSCC triggers the production of tumor-promoting and immunosuppressive cytokines IL-1 and IL-10 and results in an increased MDC migration activity. CpG-ODN is able to act contradictory to HNSCC. CONCLUSION: HNSCC is able to modulate various functions of human MDCs. CpG-ODN is suggested as a potential immunostimulatory agent of human MDC. PMID- 17465208 TI - Sequence-dependent administration of raloxifene and 5-fluorouracil/pemetrexed protects against pemetrexed cytotoxicity in human bone marrow. AB - BACKGROUND: Pemetrexed (Alimta) is a new-generation multitargeted antifolate that inhibits several key enzymes in the de novo pathways of pyrimidine and purine biosynthesis, including thymidylate synthase (TS), dihydrofolate reductase (DHFR) and glycinamide ribonucleotide formyltransferase (GARFT). Alimta has demonstrated antitumor activity in a broad array of human malignancies, e.g. breast, non-small cell lung cancer, malignant pleural mesothelioma and pancreatic, colorectal, gastric, bladder, head and neck cancer, and is currently in phase III clinical trials. It has been reported that a dose of 600 mg/m2 of pemetrexed showed toxicity to bone marrow and the gastrointestinal system. The aim of this investigation was to evaluate raloxifene (RAL) in combination with 5-fluorouracil (5-FU)/pemetrered multitargeted antifolate (MTA) to determine the most effective regimens and cellular mechanism of action to mitigate pemetrexed cytotoxicity in human bone marrow cells. MATERIALS AND METHODS: In order to determine the sequence-dependent interaction between MTA, 5-FU and RAL on proliferation, cell viability was carried out using the Quick Cell Proliferation Assay by exposing the HS-5 and MCF-7 cells to (i) MTA, 5-FU and RAL alone, or (ii) RAL 24 h prior to 5-FU followed 2 h later by MTA, or (iii) 5-FU 2 h prior to MTA followed 24 h later by RAL. RESULTS: The growth rate in MCF-7 in early RAL was 69 +/- 8.65% and late RAL was 36 +/- 4.6% of the control whereas in bone marrow early RAL was 78 +/- 8.65% and late RAL was 52 +/- 5.49% of the control. The late RAL exhibits significant protection against MTA cytotoxicity in bone marrow. The findings were further supported by cell flow cytometry, apoptosis and Western blot analysis data. CONCLUSION: This study suggests that sequence-dependent administration of RAL (5FU/MTA/RAL), in combination with 5-FU/MTA, protects against MTA toxicity in human bone marrow while maintaining the maximum inhibitory effect of pemetrexed in breast cancer. PMID- 17465209 TI - Epithelial cell nicotinic acetylcholine receptor expression in head and neck squamous cell carcinoma pathogenesis. AB - BACKGROUND: Molecular events following nicotinic acetylcholine receptor (nAChR) activation by nicotine are poorly understood. The phosphatidylinositol 3-kinase (PI3-K)/Akt/PTEN pathway has been suggested to play a role in the antiapoptotic responses to nicotine. MATERIALS AND METHODS: To elucidate the possible role of aalpha3, alpha5 and alpha7 nAChR subunit mediated PI3-K/Akt/PTEN pathway activation in squamous cell carcinoma of the head and neck (HNSCC) development, mRNA was isolated from 30 HNSCC tissues of known Akt activation state and were analyzed by reverse transcription polymerase chain reaction (RT-PCR). RESULTS: alpha3, alpha5 and alpha7 nAChR subunits were expressed in 1/30 (3.33%), 15/30 (50%) and 10/30 (33.33%), respectively. These results did not correlate with pAkt levels, previously assessed in our laboratory, or any of the clinicopathological parameters considered. CONCLUSION: This is the first report on nAChR subunit expression in human HNSCC surgical specimens of known pAkt levels. Our results suggest that nAChRs might exert their function through pathways different from PI3-K/Akt/PTEN and that alpha3, alpha5 or alpha7 nAChR subunit expression might not be useful prognostic markers in HNSCC. PMID- 17465210 TI - Non-cytotoxic and sublethal paclitaxel treatment potentiates the sensitivity of cultured ovarian tumor SKOV-3 cells to lysis by lymphokine-activated killer cells. AB - BACKGROUND: The standard treatment of epithelial ovarian cancer is tumor debulking by surgery, followed by six cycles of chemotherapy consisting of cisplatinum and paclitaxel. However, this therapy protocol is not satisfactory, since about 50% of the treated patients eventually experience recurrence within a few years of follow-up. Thus, a more innovative treatment modality is urgently needed for patients with this malignancy. We hypothesized that pretreatment of ovarian cancer SKOV-3 cells at a non-cytotoxic to sublethal dose range of paclitaxel would result in increased sensitivity to LAK-mediated killing. MATERIALS AND METHODS: MTT and trypan blue dye exclusion were used to determine the non-cytotoxic to sublethal range of paclitaxel against SKOV-3 cells. A 4-h 51Cr release cytotoxicity assay was used to evaluate the sensitivity of paclitaxel-treated and untreated SKOV-3 cells. Immunofluorescence/flow cytometric analysis was used for phenotypic changes of cells with or without paclitaxel treatment. RESULTS: Our results with trypan blue dye exclusion and MTT assays showed that the non-cytotoxic to sublethal range was between 0.001 microM and 0.01 microM. Pretreatment of SKOV-3 cells with paclitaxel at these doses for 72 h revealed significantly enhanced LAK-mediated killing against SKOV-3 cells with the highest sensitivity achieved with cells treated with 0.001 microM paclitaxel, as compared with the baseline killing of untreated cells using LAK cell alone (p < 0.05). The enhanced sensitivity of LAK-mediated killing appeared to be in part due to paclitaxel-induced expression of ICAM-1 on SKOV-3 cells. CONCLUSION: This treatment approach may be useful for further development of an effective therapeutic mode for patients with ovarian cancer. PMID- 17465211 TI - Gene expression profile of 5-fluorouracil metabolic enzymes in primary colorectal cancer: potential as predictive parameters for response to fluorouracil-based chemotherapy. AB - BACKGROUND: Thymidylate synthase (TS), dihydropyrimidine dehydrogenase (DPD), thymidine phosphorylase (TP) and orotate phosphoribosyltransferase (OPRT) have been reported to be predictive parameters for the efficacy of fluoropyrimidine based chemotherapy. Therapy guided by chemotherapy sensitivity and resistance assays may lead to rational treatment decisions. MATERIALS AND METHODS: mRNA expression of TS, DPD, TP, and OPRT were quantified by reverse-transcriptase polymerase chain reaction (RT-PCR) after harvesting from paraffin embedded specimens through microdissection. In vitro chemosensitivity testing by histoculture drug response assay (HDRA) was performed with fresh specimens of the primary tumor from 49 patients with colorectal cancer. Correlations between the results of a chemosensitivity test (the T/C ratio; the percentage of optical density of a tumor treated with anticancer drugs in relation to the optical density of the tumor cultured in RPMI 1640 medium only) and the gene expression were assessed. RESULTS: The gene expression of TS, TP, and OPRT had no correlation with clinicopathological factors, survival, and T/C ratio. The DPD mRNA levels (0.295 vs. 0.381, p = 0.2460) and OPRT/DPD ratio (5.535 vs. 4.394. p = 0.226) had a weak correlation with the T/C ratio. Of the eleven patients who were actually treated with chemotherapy, the responders had higher OPRT/DPD ratios (8.065 vs. 4.081, p = 0.090). CONCLUSION: The DPD mRNA level and OPRT/DPD ratio evaluated from paraffin embedded specimens are candidates for further evaluation as predictors of response against 5-fluorouracil-based chemotherapy in colorectal cancer. PMID- 17465212 TI - Induction of apoptosis by morphine in human tumor cell lines in vitro. AB - Most previous studies of the induction of tumor cell apoptosis by morphine have been conducted with concentrations very much higher than those used clinically. An investigation of the ability of morphine to induce apoptosis at its clinical concentration (10(-8) M) was therefore undertaken. Cytotoxicity was tested by 3 [4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay, induction of early apoptosis and necrosis by fluorescence-activated cell sorter (FACS) analysis with Annexin V and propidium iodide (PI), activation of caspase -2, -3, 8 and -9 by cleavage of specific substrates, DNA fragmentation by agarose gel electrophoresis, radical intensity and O2- scavenging activity by ESR spectroscopy. Millimolar concentrations of morphine showed higher cytotoxicity against human tumor cell lines (HL-60, A549, MCF7) than against normal human cells (HGF, HPC, HPLF). The clinical concentration of morphine produced early apoptotic markers in HL-60 and A549 cells whereas it induced higher numbers of necrotic cells in MCF7 cells, both in a naloxone-sensitive manner. The clinical concentration of morphine failed to activate any caspase species and induced only trace amounts of internucleosomal DNA fragmentation, in contrast to cytotoxic concentrations of morphine. Morphine, with a C-3 hydroxyl group, showed higher cytotoxicity and O2- scavenging activity than codeine, in which the hydroxyl group at C-3 was replaced with a methoxy group, suggesting the involvement of a radical-mediated reaction. The present report may offer new strategies for treatment and prevention of cancer using a clinical concentration of morphine not only as an anti-nociceptive, but also as an apoptosis or necrosis inducer. PMID- 17465213 TI - CPT-11 (SN-38) chemotherapy may be selectively applicable to biliary tract cancer with low hMLH1 expression. AB - Biliary tract cancer is of highly malignancy with a poor 5-year survival. However, established chemotherapeutic regimens have not yet been established. Previously, we have reported that hMLH1, a mismatch repair (MMR) gene was frequently (57%) found to be lacking in surgically resected biliary tract carcinomas and the patients lacking the expression of hMLH1 revealed a poorer prognosis than those patients who possessed it. The MMR gene has been considered to be associated with sensitivity to various chemotherapeutic agents that act on DNA. A loss of MMR expression has been reported to increase sensitivity to topoisomerase inhibitors such as etoposide (ETP) or camptothecins (CPT). In the present study, whether or not hMLH1 deficiency resulted in a higher sensitivity to irinotecan (CPT-11) active form (SN-38) was investigated using a short interfering (Si)RNA system. A quantitative reverse transcription-polymerase chain reaction (RT-PCR) was conducted to measure the levels of hMLH1 expression in seven cancer cell lines, and this was compared with the drug sensitivity (IC50) to SN-38. The hMLH1 expression was correlated with the IC50 for SN-38, although the relationship was not statistically significant (R = 0.717, p = 0.0715). SiRNA double strand RNA (dsRNA) was transiently transfected into KMG-C (gallbladder cancer) cells. hMLH1 mRNA expression was repressed by hMLH1 dsRNA in a dose dependent manner in comparison to the control dsRNA. The cell growth of the hMLH1 dsRNA transfected group was decreased by approximately 50% by SN-38 exposure. Flow cytometry was also carried out to examine the effect of the SN-38 treatment on the cell cycle. Following hMLH1 dsRNA transfection, the subG1 fraction was increased in comparison with the control in a dose-dependent manner. In conclusion, a low expression of hMLH1 in biliary tract cancer may aid in predicting its responsiveness to CPT-11 (SN38). PMID- 17465214 TI - Evaluation of cell death caused by an ethanolic extract of Serenoae repentis fructus (Prostasan) on human carcinoma cell lines. AB - BACKGROUND: Phytotherapy is a third approach for treating lower urinary tract symptoms associated with benign prostatic hyperplasia (BPH). The lipido-sterolic extract of the fruit of Serenoa repens is one of the more widely used phytotherapeutic agents in this regard. MATERIALS AND METHODS: The effect of an ethanolic extract of S. repens (10-1000 microg/ml) was tested in hormone sensitive LNCaP, MCF-7 and hormone-insensitive DU 145, MDA MB231 prostate, breast carcinoma cell lines, renal Caki-1, urinary bladder J82, colon HCT 116 and lung A 549 cancer cells. Its cell growth inhibitory and apoptosis-inducing effects were tested using WST-1 assay and flow cytometry (Annexin V/PI stain) and/or by colorimetric assay (APOPercentage assay). RESULTS: The S. repens extract induced a dose-dependent antiproliferative effect on all human malignant cells tested, with GI50 values between 107 and 327 pmicro/ml. In hormone-sensitive prostate LNCaP and breast MCF-7 cell lines, the effect of extract expressed in GI50 was 2.2- and 2.5-fold more potent (p < 0.01) than in hormone-insensitive DU 145 and MDA MB231 cells. The proportion of apoptotic cells, except in A549 cells, lay between 22.5-36.3%. S. repens extract did not induce apoptosis in lung cancer A 549 cells. CONCLUSION: This study showed that the antiproliferative effect exerted by the ethanolic extract of S. repens is at least triggered by induction of apoptosis. These in vitro data provide some information that may be useful for clinical use and render S. repens extract an interesting tool for new applications. PMID- 17465215 TI - Structure-activity relationship studies of arylthiazolidine amides as selective cytotoxic agents for melanoma. AB - New drugs are urgently needed for improved therapy for melanoma. MATERIALS AND METHODS: Ninety-one novel compounds were evaluated in two melanoma and one normal skin cell lines to identify potential lead compounds with high potency and selectivity. Mechanisms of action for the best compound were also investigated. RESULTS: Three potent lead structures (serine amino alcohols, serine amides and thiazolidines) were identified, with thiazolidines having both excellent potency and high selectivity when compared with sorafenib, a drug used extensively in clinical trials for melanoma. Analyzing the effect of the lead compound showed that it induced DNA degradation consistent with necrotic cell death. CONCLUSION: The lead structure represents a novel class of compounds that can be further optimized for potential drug to treat advanced melanoma. PMID- 17465216 TI - EGFR and VEGFR2 protein expressions in bone metastases of clear cell renal cancer. AB - BACKGROUND: EGFR and VEGFR2 protein expressions are hallmarks of clear cell renal cancer (RCC) with questionable prognostic impact. The skeletal system is one of the most common metastatic sites of RCC. Unfortunately, there are no data for EGFR and VEGFR2 protein expression in such lesions. MATERIALS AND METHODS: Twenty cases of bone metastatic clear cell RCC were analyzed. EGFR and VEGFR2 proteins were detected by immunohistochemistry and analyzed by morphometry scoring both % positivity and the intensity. RESULTS: EGFR protein scores were significantly reduced in bone metastases of RCC due to the reduction of EGFR protein expression in about one third of the cases (7/20). The VEGFR2 protein-positive phenotype of clear cell RCC was relatively frequent (7/20, 35%), but was lost in bone metastases (2/20, 10%). CONCLUSION: These data suggest a phenotypic/genotypic change of clear cell RCC during the progression to bones and warrant further investigation. PMID- 17465217 TI - Prohibitin in the pathogenesis of transitional cell bladder cancer. AB - Our profiling experiment demonstrated that prohibitin 1 (PHB) was ubiquitously expressed in uroepithelial and urothelial carcinoma cell lines and exhibited a trend toward a positive relationship with tumor progression. The aim of this study was, therefore, to examine the potential role of PHB in multistage bladder carcinogenesis and predicting patient outcome. Immunohistochemical staining showed that PHB was overexpressed in 141 out of 167 cases (84.4%) of bladder cancer. This expression was positively related to met receptor overexpression (p = 0.04) and to multiple tumors (p = 0.05). Independent factors in predicting patient survival were multiple tumors (p = 0.002), muscle invasion (p = 0.003), and met overexpression (p = 0.05) in a multivariate analysis. Interestingly, patients with superficial bladder cancer overexpressing both PHB and met had a significantly lower recurrence-free survival rate than those not expressing PHB (p = 0.04). Taken together, our findings showed that PHB was activated at an early stage of carcinogenesis and that it may play a synergistic role with met in the progression of human bladder cancer. In addition, we demonstrated that genistein and justicidin A, natural chemoprevention agents, could suppress the expression of PHB in vitro. Thus, targeting PHB would be a useful approach for treating and preventing human bladder cancer. PMID- 17465218 TI - [Methyl-3H]-choline incorporation into MCF-7 cells: correlation with proliferation, choline kinase and phospholipase D assay. AB - BACKGROUND: [Methyl-3H]-choline is a promising new positron emission tomography (PET) agent used for cancer imaging whose mechanism has still not fully been elucidated. In this study, whether [methyl-3H]-choline determined by measuring the activity of choline kinase (ChoK) and phospholipase D (PLD) in rapidly proliferating and confluent breast cancer MCF-7 cells is related to cell proliferation or not was investigated. MATERIALS AND METHODS: The activity of ChoK and PLD were determined using ion exchange chromatography and transphosphatidylation assay respectively. RESULTS: [Methyl-3H]-PCho content expressed as pmol mg(-1) protein(-1) min(-1) (n = 6) was significantly higher in the exponentially growing (484.04 +/-2 0.23) compared with confluent (70.35 +/ 9.83) cells using Student's t-test (p < 0.001). Moreover, PLD activity expressed as the mean (n = 6) (disintegration per minute (d.p.m.)/microg protein +/- SD (mean S phase +/- SD)) showed significantly higher (p < 0.001) activity in the exponentially growing cells (196.39 +/- 2.21 d.p.m./microg protein (39.69 +/- 4.00%)) compared with confluent cells (99.10 +/- 1.35 d.p.m./microg protein (9.33 +/- 0.82%)). CONCLUSION: This study indicates that the major water-soluble choline metabolite was phosphocholine (PCho) as a consequence of increased ChoK and PLD activity in the exponentially growing cells compared to confluent cells. PMID- 17465219 TI - Biodistribution and elimination characteristics of two 111In-labeled CCK 2/gastrin receptor-specific peptides in rats. AB - BACKGROUND: Due to their high CCK-2/gastrin receptor selectivity, high affinity, and rapid background clearance, radiolabeled minigastrins (MG) are emerging as promising new tools in the diagnosis and therapy of CCK-2/gastrin receptor positive tumors. In this study, the pharmacokinetic profile, particularly the excretion mode, of two 111In-labeled minigastrins was compared in rats. The first tracer, 111In-MG-0 is based on (D)Glu1-MG, while the second, 111In-MG-11, is its des-(Glu)5-derivative, expected to be less retained in renal tissue. MATERIALS AND METHODS: The fate of 111In-MG-0 and 111In-MG-11 in the body of rats was investigated during biodistribution and bioelimination experiments, while the respective elimination parameters were determined in perfused rat liver and kidney models. RESULTS: During biodistribution both compounds were rapidly cleared from the blood and most non-target organs whereas activity levels in the bowel and stomach declined slowly. The overall contribution of hepatobiliary excretion of 111In-MG-0 and 111In-MG-11 was relatively small. In the perfused rat liver their elimination into the bile was negligible. In contrast, renal excretion was the major excretion pathway for both analogs, mainly via glomerular filtration. However, kidney levels were substantially higher and retention was more prolonged in the case of 111In-MG-0 as compared to 111In-MG-11. CONCLUSION: The presence of the (Glu)5-chain in 111Ln-MG-0 appears to be implicated in the prolonged radioactivity retention in the kidney of rats. PMID- 17465220 TI - Expression of the HER family mRNA in breast cancer tissue and association with cell cycle inhibitors p21(waf1) and p27(kip1). AB - BACKGROUND: The HER family of the receptor tyrosine kinases epidermal growth factor receptor (EGFR), HER2, HER3 and HER4 are involved in the pathogenesis of many human malignancies. Although there is extensive literature on the expression of single HER-2 and EGFR receptors in breast cancer, little is known concerning the simultaneous expression at the mRNA level of these four receptors in human breast tissue and their influence in downstream signaling pathways that control cell cycle and proliferation. MATERIALS AND METHODS: The mRNA expression pattern of the four HER-receptors has been investigated and correlated with the expression of the cyclin-dependent kinase (CDK) inhibitors p21(Waf1) and p27(Kip1) in 67 breast cancer specimens. RESULTS: A positive correlation between HER-3 and HER-4 mRNA levels and a negative correlation between HER-2 and HER-3 was found. Compared to normal breast tissue, all four receptors were overexpressed in breast tumors and the strongest overexpression was found for HER 3 (p = 0.001). HER-4 expression was inversely related to histopathological grading (HPG), suggesting that elevated HER-4 mRNA expressions could be a biological marker of a more differentiated phenotype. The expression of p21(Waf1) protein was higher in HER-2-negative tumors, compared to HER-2-positive breast carcinomas. Compared to normal breast tissue, p21delta, the 19 kDa degraded form of p21 protein, was markedly expressed in breast cancer (p < 0.001). Conversely, p27(Kip1) was positively associated with HER-2 receptor and inversely associated with HER-3. CONCLUSION: The HER family members are overexpressed in breast cancer. Complex patterns of HER family expression were observed and the effect on cell cycle regulation was dependent on that pattern. PMID- 17465221 TI - Methylation-mediated silencing of genes is not altered by selenium treatment of prostate cancer cells. AB - BACKGROUND: The role of selenium in reducing the risk of multiple cancers has been described in the literature. Although reports have described the antiproliferative and pro-apoptotic function of selenium by up-regulation of genes in these pathways, information is lacking on the target mechanisms of selenium on specific genes. This study examines whether selenium treatment alters the methylation status of epigenetically silenced genes in prostate cancer cells. MATERIALS AND METHODS: Methylation of glutathione sulfotransferase pi (GSTP1) and Ras associated family 1A (RASSF1A) genes was studied using methylation sensitive PCR (MS-PCR). Gene expression was studied using Reverse Transcriptase PCR and Western Blotting. RESULTS AND CONCLUSION: Treatment of prostate cancer cells with selenium did not alter the expression of genes that were silenced by DNA methylation. Furthermore, the methylation status of these genes remained unaltered after treatment with seleno-DL-methionine. PMID- 17465222 TI - Incadronate induces cell detachment and apoptosis in prostatic PC-3 cells. AB - BACKGROUND: Bisphosphonates are widely used for the treatment and prevention of osteoporosis and are also effective in the treatment of bone metastasis of prostate cancer. Several mechanisms underlying the antitumor effect of bisphosphonates have been proposed, including direct effects on tumor cells, such as induction of apoptosis and inhibition of invasion. MATERIALS AND METHODS: The detached and adherent cells after incadronate treatment were collected separately and stained with trypan blue solution. RESULTS: It was found that incadronate induced cell detachment with dephosphorylation of focal adhesion kinase (FAK). The induction of cell detachment by incadronate was prevented by coincubation with geranylgeraniol. The activation of caspase-3 was observed in incadronate treated floating cells, but not in the adherent cells. A caspase inhibitor did not inhibit cell detachment by incadronate but it markedly prevented cell death. CONCLUSION: These results suggest that incadronate induces cell detachment, followed by caspase-dependent apoptosis. PMID- 17465223 TI - Effects of ACE I/D polymorphism on prostate cancer risk, tumor grade and metastatis. AB - The aim was to substantiate the putative significance of angiotensin-converting enzyme (ACE) (insertion/deletion) I/D polymorphism on prostate cancer risk, BTPSA A TPSA (before treatment-after treatment prostate-specific antigen) levels and tumor development. MATERIALS AND METHODS: 48 prostate cancer patients and 51 healthy volunteers were included. The ACE I/D genotypes were determined by PCR (polymerase chain reaction) and RFLP (restriction fragment length polymorphism) techniques. RESULTS: The DD genotype may have detrimental and the II genotype may have protective effect on prostate cancer (p = 0.03). The highest before treatment PSA (BTPSA) values were found in the patient group having the DD genotype (p = 0.017). PSA-AT levels were higher in homozygous mutant DD than homozygous II and the decrease in PSA-AT level was found to be statistically significant in each genotype (p = 0.000). Patients with the D allele showed a higher prevalence of late stage prostate carcinoma when compared to the patients with II genotype (p = 0.022) and the detrimental effects of the D allele, both in lymph node metastases and distant metastasis were observed. CONCLUSION: The risk of prostate cancer development, the PSA level and tumor metastasis may be associated with genetic variation in the ACE I/D genotypes which may be used as an important biomarker for further studies. PMID- 17465224 TI - Inhibition of cancer cell proliferation and suppression of TNF-induced activation of NFkappaB by edible berry juice. AB - BACKGROUND: Berries contain several phytochemicals, such as phenolic acids, proanthocyanidins, anthocyanins and other flavonoids. There has been growing interest in a variety of potential chemopreventive activities of edible berries. The potential chemopreventive activity of a variety of small berries cultivated or collected in the province of Quebec, Canada were evaluated here. MATERIALS AND METHODS: Strawberry, raspberry, black currant, red currant, white currant, gooseberry, high-bush blueberry, low-bush blueberry, velvet leaf blueberry, serviceberry, blackberry, black chokeberry, sea buckthorn and cranberry were evaluated for antioxidant capacity, anti-proliferative activity, anti inflammatory activity, induction of apoptosis and cell cycle arrest. RESULTS: The growth of various cancer cell lines, including those of stomach, prostate, intestine and breast, was strongly inhibited by raspberry, black currant, white currant, gooseberry, velvet leaf blueberry, low-bush blueberry, sea buckthorn and cranberry juice, but not (or only slightly) by strawberry, high-bush blueberry, serviceberry, red currant, or blackberry juice. No correlation was found between the anti-proliferative activity of berry juices and their antioxidant capacity (p > 0.05). The inhibition of cancer cell proliferation by berry juices did not involve caspase-dependent apoptosis, but appeared to involve cell-cycle arrest, as evidenced by down-regulation of the expression of cdk4, cdk6, cyclin D1 and cyclin D3. Of the 13 berries tested, juice of 6 significantly inhibited the TNF induced activation of COX-2 expression and activation of the nuclear transcription factor NFkappaB. CONCLUSION: These results illustrate that berry juices have striking differences in their potential chemopreventive activity and that the inclusion of a variety of berries in the diet might be useful for preventing the development of tumors. PMID- 17465225 TI - Overexpression of the human homologue for Caenorhabditis elegans cul-4 gene is associated with poor outcome in node-negative breast cancer. AB - BACKGROUND: Cul-4, a member of the Caenorhabditis elegans "cullin" ubiquitin ligase gene family, plays a critical role in regulation of DNA-replication in this nematode. It has been suggested that cul-4 might have an important role in the development and progression of human cancer, but no data on this subject exist. The aim of this study was to investigate the expression and prognostic relevance of CUL-4 protein in lymph node-negative breast cancer, one of the most common malignancies worldwide. MATERIALS AND METHODS: CUL-4 protein expression was determined with immunohistochemistry in 167 specimens of human node-negative invasive breast cancer with long-term follow-up. Results were correlated with overall and disease-free survival of patients. RESULTS: Strong expression of CUL 4 protein was observed in 32 cases (19.2%), moderate expression in 59 (35.3%), weak expression in 64 (38.3%), and in 12 tumors (7.2%) no expression of CUL4 was observed. Patients with strong expression of CUL4 had a significantly shorter overall and disease-free survival (p = 0.04 and p = 0.029, respectively; Cox regression) compared to all other cases. CONCLUSION: Our data provide evidence for the first time that CUL-4 could play an important role in the development and progression of human cancer. PMID- 17465226 TI - Sodium phenylacetate (NaPa) improves the TAM effect on glioblastoma experimental tumors by inducing cell growth arrest and apoptosis. AB - BACKGROUND: Multiform glioblastomas represent the most aggressive brain tumors. Here, the cooperative effects of sodium phenylacetate (NaPa) and/or tamoxifen (TAM) on CNS1 and 9L glioblastoma cell lines in vitro and in an experimental animal tumor model were investigated. MATERIALS AND METHODS: The drug effects on cell cycle and apoptosis were investigated by flow cytometry. CNS1 cells were implanted subcutaneously in nude mice to form tumors which were then treated with NaPa, TAM or NaPa/TAM. RESULTS: A significant inhibitory effect of NaPa on the two glioma cell lines (LD50 of 10 mM) was observed. 10(-5) M of TAM inhibited approximately 35% of 9L cell growth, and 90% of CNS1 cell growth. When a combination of both drugs included 10(-9) M of TAM, inhibition of about 50% of 9L cell growth and 75% of CNS1 cell growth occurred. The NaPa/TAM combined treatment increased the number of G0/G1 arrested cells and apoptotic cells as compared to treatments with NaPa or TAM alone. Inhibition of CNS1 tumor growth were observed after a two week treatment with NaPa (32 mg/kg/day) or TAM (6 mg/kg/day). CONCLUSION: These results showed a synergistic effect between these two drugs on tumor cell proliferation, caused by cell cycle arrest in the G0/G1 phase and by induction of apoptosis. PMID- 17465227 TI - The prognostic significance of human epidermal growth factor receptor correlations in squamous cell cervical carcinoma. AB - The aim of this study was to investigate the expression and prognostic influence of HER1 (EGFR), HER2 (c-erb-B2), HER3 (c-erb-B3) and HER4 (cerb-B4) in squamous cell cervical carcinomas (SCC) and the importance of receptor correlations. PATIENTS AND METHODS: 78 SCC were stained immunohistochemically for HER1-HER4. HER2 gene amplification was determined using fluorescence in situ hybridization (FISH). Parametric correlations were performed between the four receptors and tumor characteristics. Overall survival was evaluated by uni- and multivariate analyses. RESULTS: Overexpression was found in 63% of SCC for HER1, in 21.8% for HER2, in 74.4% for HER3 and in 79.5% for HER4. Correlations were observed between HER1 and HER4 (p = 0.019). Survival analyses revealed a significant association of HER1 overexpression with favorable outcome (p = 0.016), while overexpression of HER2 and HER3 was associated with poor prognosis (p = 0.006; p = 0.05, respectively). HER1 remained significant in multivariate analysis. CONCLUSION: These data suggest that the prognostic relevance of the different HER receptors is influenced by the balance between the various receptors, especially of HER4. PMID- 17465228 TI - Regulation of p53-, Bcl-2- and caspase-dependent signaling pathway in xanthorrhizol-induced apoptosis of HepG2 hepatoma cells. AB - Xanthorrhizol is a sesquiterpenoid compound extracted from the rhizome of Curcuma xanthorrhiza. This study investigated the antiproliferative effect and the mechanism of action of xanthorrhizol on human hepatoma cells, HepG2, and the mode of cell death. An antiproliferative assay using methylene blue staining revealed that xanthorrhizol inhibited the proliferation of the HepG2 cells with a 50% inhibition of cell growth (IC50) value of 4.17 +/- 0.053 microg/ml. The antiproliferative activity of xanthorrhizol was due to apoptosis induced in the HepG2 cells and not necrosis, which was confirmed by the Tdt-mediated dUTP nick end labeling (TUNEL) assay. The xanthorrhizol-treated HepG2 cells showed typical apoptotic morphology such as DNA fragmentation, cell shrinkage and elongated lamellipodia. The apoptosis mediated by xanthorrhizol in the HepG2 cells was associated with the activation of tumor suppressor p53 and down-regulation of antiapoptotic Bcl-2 protein expression, but not Bax. The levels of Bcl-2 protein expression decreased 24-h after treatment with xanthorrhizol and remained lower than controls throughout the experiment, resulting in a shift in the Bax to Bcl-2 ratio thus favouring apoptosis. The processing of the initiator procaspase-9 was detected. Caspase-3 was also found to be activated, but not caspase-7. Xanthorrhizol exerts antiproliferative effects on HepG2 cells by inducing apoptosis via the mitochondrial pathway. PMID- 17465229 TI - P53 expression is associated with malignant potential in xenograft tissues of a fibrosarcoma mouse model. AB - BACKGROUND: The expression of wild-type and mutant p53 was studied in two fibrosarcoma cell lines in a mouse xenograft model. MATERIALS AND METHODS: Human cell lines HT1080 and Hs913(D)T were implanted in athymic mice via intramuscular (i.m.) or subcutaneous (s.c.) routes. After eight weeks, liver, lung and primary inoculation sites were harvested. Sections were stained using two methods: a) haematoxylin and eosin to detect tumour at implantation site, liver and lung; b) immunohistochemistry using monoclonal antibodies to detect expression of wild type (wt) and mutant p53. RESULTS: Both cell lines had similar implantation rates via either route but Hs913(D)T had a higher metastatic rate than HT1080. The Hs913(D)T cells exhibited greater expression of mutant and wild-type p53 than the HT1080 cells. CONCLUSION: The expression of wild-type and mutant p53 is associated with a cell line of greater malignant potential. The inoculation route does not affect primary tumour uptake or metastatic rate. PMID- 17465230 TI - Loss of tumour suppressor p16 expression in initial stages of oral oncogenesis. AB - BACKGROUND: The p16 tumour suppressor gene is known to be involved in regulation of the cell cycle. p16 expression in sequential histological stages of oral squamous cell carcinoma (OSCC) formation was investigated using an experimental model of induced oral carcinogenesis in Syrian golden hamsters. MATERIALS AND METHODS: Thirty-seven animals were divided into one control group (N = 7) and three experimental groups (N = 10 each) which were treated with a carcinogen and sacrificed at 10, 14 and 19 weeks after treatment. Tumour sections were studied immunohistochemically using monoclonal antibodies against p16 protein. RESULTS: p16 was found significantly increased in hyperplasia, sharply decreased in dysplasia and in the subsequent stages of oral carcinogenesis. CONCLUSION: Inactivation of p16 occurs at the early stage of oral mucosal dysplasia in the multistep process of oral tumourigenesis. Therefore, p16 may be considered as a useful prognostic marker for the progression of oral cancer. PMID- 17465231 TI - Locoregional IL-2 therapy in the treatment of colon cancer. Cell-induced lesions of a murine model. AB - BACKGROUND: Local therapy with IL-2 may be very effective in the treatment of different forms of cancer. The aim of this study was to determine the effectiveness of IL-2 locoregional application in the treatment of colon cancer. MATERIALS AND METHODS: Twenty eight syngenic BDIX rats were utilized in this study. The rats were divided into two groups of fourteen animals: group T (treatment) and group C (control). All rats of both groups were injected, under the splenic capsule, with T 10(7) DHD/K2/ TRb neoplastic cells. Then, within and around the site of the previous inoculation, the T group was injected with 1 ml of glucosate solutions + 0.1% albumin (BSA) containing 2.5 x 10(6) IU of IL-2 ( Proleukin-Chiron), whereas the C group was injected with 1 ml of BSA alone. After three weeks, rats were sacrificed and the liver and spleen were removed. The following parameters were considered: volume and weight, neoplastic-non neoplastic tissue index of the spleen, mitotic index and vascular density of splenic and hepatic lesions. RESULTS: All the studied parameters showed statistically significant differences in treated and untreated animals. CONCLUSION: This study of a murine model demonstrated that IL-2 locoregional therapy may be effective in the treatment of colon cancer. PMID- 17465232 TI - Fas-1377A/G and FasL-844 T/C gene polymorphisms and epithelial ovarian cancer. AB - BACKGROUND: The Fas receptor is known to be widely expressed in various tissues and FasL is highly expressed on cells of the immune system and also on cells of immune-privileged areas such as the eyes and brain. Ovarian cells are known to exhibit marked FasL immunoreactivity throughout follicular development; there may also be a relationship between Fas and FasL polymorphisms and the immune privileges of the epithelial ovarian cells. PATIENTS AND METHODS: The study included 47 epithelial ovarian carcinoma patients and 41 healthy subjects. Polymerase chain reaction (PCR) and restriction endonucleases were used to determine the polymorphic Fas and FasL genes. RESULTS: The FasL CC genotype was found to increase the risk of ovarian carcinoma and a protective effect of the GGCT genotype was observed. CONCLUSION: Because of the expressional aspects of the FasL-844T --> C polymorphism, individuals carrying the FasL-844C allele would be expected to have higher FasL expression on tumour cells compared with those carrying the FasL-844T allele. People with such a genotype show a tendency to develop various tumours. PMID- 17465233 TI - Combination of cytotoxic-differentiation therapy with 5-fluorouracil and phenylbutyrate in patients with advanced colorectal cancer. AB - BACKGROUND: Phenylbutyrate (PB), a histone deacetylase inhibitor (HDACi), has been shown in laboratory studies to potentiate growth inhibition by 5 fluorouracil (FUra) of human colon carcinoma cells. PATIENTS AND METHODS: Phase I trial of FUra (24-hour continuous intravenous infusion (CIV)) with dose escalation (2 g/m2 to 2.3 g/m2), in combination with PB (120 hour CIV at fixed dose 410 mg/kg/d x 5), repeated weekly, in patients with advanced colorectal cancer. RESULTS: Nine patients with metastatic colorectal cancer were treated, 8 of whom were evaluable for toxicity. Toxicities were dose-dependent, reversible and included somnolence, fatigue, confusion, hearing loss, triglyceridemia and hyperuricema. Three out of 4 patients who completed at least 8 weeks of treatment had stable disease (SD) lasting 12+, 25 and 54 weeks (2 out of the 3 patients with SD have had multiple prior chemotherapy regimens). CONCLUSION: Weekly infusions of FUra followed by PB were fairly well tolerated with disease stabilization in 3/4 (75%) of patients. This is the first report to demonstrate the feasibility of combining a cytotoxic agent with a HDACi as a cancer treatment. PMID- 17465234 TI - Multicenter phase II study of irinotecan plus bolus fluorouracil/l-leucovorin for metastasic colorectal cancer. AB - Treatment of metastatic colorectal cancer remains inadequate. PATIENTS AND METHODS: In a multicentre Phase II study, irinotecan (100 mg/m2), 5-fluorouracil (5-FU) (500 mg/m2), and l-leucovorin (l-LV) (250 mg/m2) were administered on days 1, 8, and 15 of a five-week cycle. Forty-five patients were enrolled. RESULTS: The objective response rate was 26.7%. The median survival time was 21.8 months and the one-year survival rate was 73.3%. The median number of cycles was 4.0, with a median relative dose intensity of 83.3% for both irinotecan and 5-FU. Grade 3 or 4 haematological toxicities were anaemia in four patients, leukopaenia in six patients, and neutropaenia in 15 patients, while non-haematological toxicities were diarrhoea in three patients, and nausea, vomiting, anorexia and increased transaminases in two patients each. No treatment-related deaths occurred. CONCLUSION: Irinotecan plus 5-FU/l-LV can be used to treat metastatic colorectal cancer on an outpatient basis. PMID- 17465235 TI - Dosage of capecitabine and cyclophosphamide combination therapy in patients with metastatic breast cancer. AB - BACKGROUND: Capecitabine is a highly effective and well-tolerated treatment for metastatic breast cancer (MBC) and extends survival when combined with docetaxel. Capecitabine and cyclophosphamide are orally administered and have preclinical synergy and non-overlapping toxicities. PATIENTS AND METHODS: Sixteen pretreated MBC patients received escalating doses of oral capecitabine 628 to 829 mg/m2 twice daily (bid) plus oral cyclophosphamide 33 to 50 mg/m2 bid, on days 1 to 14 every 21 days. RESULTS: Among the ten patients receiving capecitabine/cyclophosphamide 829/33 mg/m2 bid on days 1 to 14, two experienced dose-limiting toxicities (DLT, treatment delay > 1 week due to grade 2 leukopenia). Because neither patient developed further grade > 1 toxicity and none of the patients experienced grade 3/4 toxicities or further DLTs, this dose level is the recommended regimen, producing partial responses in two of five evaluable patients. CONCLUSION: The recommended all oral capecitabine/cyclophosphamide combination regimen is well tolerated and active in MBC, and is being evaluated in a phase II study in anthracycline-pretreated MBC. PMID- 17465236 TI - Ultrasonographic-guided pervaginal cul-de-sac cytology in the follow-up of ovarian carcinoma. AB - The aim of this study was to compare, prospectively, traditional pervaginal cul de-sac aspiration cytology with an ultrasonographic-guided aspirate in the detection of residual or recurrent ovarian carcinoma. PATIENTS AND METHODS: Fifty one patients with ovarian carcinoma were monitored during chemotherapy (21 patients) or follow-up (30 patients) after first-line treatment. All patients underwent both traditional blind pervaginal cul-de-sac aspiration cytology and an ultrasonographic-guided pervaginal aspirate. The samples were classified as class 0 or insufficient when no mesothelial cells were detected in the aspirate. The results of cytological classification of the aspirates were compared with each other according to sampling order. RESULTS: Samples were classified as class 0 in 56% when the traditional cul-de-sac aspiration was taken first, and in 73% when ultrasonographic-guided aspiration was taken first (p = 0.249, Fisher's exact test). The number of class 0 samples was smaller among those taken second than among those taken first (22 (44%) vs. 33 (65%), p = 0.046). Four recurrences were detected during the mean follow-up of six months (range 2-11 months) in 30 patients who were followed-up after the first-line treatment. In one case, a positive cul-de-sac cytology was the first and only early indication of recurrence. CONCLUSION: The use of ultrasonography did not improve the accuracy of the cul-de-sac aspiration. The greater amount of fluid in the cul-de-sac during the second sampling might contribute to achieving a better result. PMID- 17465237 TI - A phase I study of cisplatin, temozolomide and thalidomide in patients with malignant brain tumors. AB - BACKGROUND: There is pre-clinical evidence of synergism between cisplatin (P) and temozolomide (T) due to higher inhibition by T of O6-alkyl-guanine alkyltransferase (AGAT), an enzyme involved in the mismatch repair system. T and P are active against malignant gliomas while thalidomide (TH) is emerging as an inhibitor of angiogenesis. PATIENTS AND METHODS: Triplets of patients with malignant brain tumors received escalating doses of P, T and TH up to the dose limiting-toxicity (DLT) and the maximal tolerated dose (MTD). RESULTS: Seventeen patients were enrolled and a total of 74 cycles were delivered. The MTD was P 75 mg/m2 day 1 and T 150 mg/m2 days 1 to 5 every 21 days with a concomitant 200 mg total daily dose of TH. DLT events were G4 thrombocytopenia and febrile neutropenia. CONCLUSION: Concomitant administration of P 75 mg/m2 day 1, T 150 mg/m2 days 1 to 5 every 21 days and concomitant TH at a total daily dose of 150 mg is feasible and safe. Early efficacy data are encouraging and a phase II study is ongoing. PMID- 17465238 TI - The prognostic role of the STK15 T91A polymorphism and of STK15 mRNA expression in patients with urothelial cell carcinoma. AB - BACKGROUND: The prognostic role of the STK15 T91A polymorphism and of STK15 mRNA expression was investigated in patients with urothelial cell carcinoma (UCC). MATERIALS AND METHODS: The STK15 genotype with respect to the T91A polymorphism was assessed by restriction fragment length polymorphism in 135 patients. STK15 mRNA expression was measured in tumor tissues of 103 patients, using real-time quantitative PCR. RESULTS: The T91A polymorphism lacked any prognostic information in our patient cohort. Interestingly though, STK15 mRNA expression was increased in invasive and high-grade tumors (p-values of 0.009 and 0.0001, respectively). Additionally, patients with superficial UCC (n = 82) who had a tumor recurrence in the first year after surgery displayed elevated STK15 mRNA expression levels (p = 0.009). Kaplan-Meier survival analysis revealed an increased risk of tumor progression for patients with Ta tumors (n = 62) and high STK15 expression (log-rank p = 0.04). Furthermore, a decreased overall (log-rank p = 0.006) and UCC-specific survival (log-rank p = 0.001) were shown for patients with elevated STK15 mRNA levels. CONCLUSION: Patients with UCC and elevated levels of STK15 mRNA generally showed a more adverse disease course than patients with low levels. This may help in identifying patients in need of more aggressive treatment. PMID- 17465239 TI - Impact of tumor biological factors on response to pre-operative epirubicin and paclitaxel chemotherapy in primary breast cancer. AB - BACKGROUND: The objective of this study was to determine the predictive impact of several established tumor biological factors (progesterone receptor, estrogene receptor, HER2/neu, and Ki-67) on response to pre-operative combination chemotherapy with epirubicin and paclitaxel in a representative group of primary breast cancer patients. PATIENTS AND METHODS: Thirty-eight primary breast cancer patients (metastasis-negative) received pre-operative chemotherapy with epirubicin and paclitaxel. The response characteristics analyzed included pathological complete response and partial remission determined by 3D ultrasound, as well as down-staging and breast conserving surgery. RESULTS: Pathologically complete response occurred in six patients. Overexpression of HER2/neu, low (negative) hormone receptors and high Ki-67 were all significant positive predictive factors for response to pre-operative chemotherapy. CONCLUSION: HER2/neu overexpression is predictive for response not only to trastuzumab, but also to epirubicin and paclitaxel. PMID- 17465240 TI - Long-term follow-up for patients with papillary thyroid carcinoma treated as benign nodules. AB - BACKGROUND: The recent prevalence of ultrasonography and fine-needle aspiration biopsy (FNAB) has facilitated the detection and diagnosis of papillary thyroid carcinoma. However, there are still cases that are preoperatively misdiagnosed and treated as benign nodules because ultrasonographic and FNAB findings do not provide sufficient evidence for a malignant diagnosis. In this study, we investigated the clinical outcomes of patients with papillary carcinoma that had not been recognized preoperatively. PATIENTS AND METHODS: We investigated the prognoses of 56 patients with papillary carcinoma who underwent thyroidectomy without node dissection under a diagnosis of benign nodules. RESULTS: None of the patients underwent further surgery such as completion total thyroidectomy and node dissection after the pathological diagnosis of papillary carcinoma was established. However, to date, only 3 patients (5.3%) showed recurrence 116, 133 and 148 months after the initial surgery, respectively. Two patients showed recurrence in the remnant thyroid and one showed recurrence in the bone. None of the patients have died of thyroid carcinoma. CONCLUSION: Papillary carcinomas misdiagnosed as benign nodules on ultrasonography and FNAB are indolent and very slow-growing. Immediate further surgery is not needed for such cases, even if they were resected as benign nodules at the initial surgery. PMID- 17465241 TI - 32-year survival with metastatic adrenal cortical carcinoma--update of a case report. AB - Adrenal cortical carcinoma (ACC) is a rare and highly malignant tumour with up to 70% of the patients diagnosed at an advanced clinical stage and up to 40% presenting with metastasis. In 2004, we reported a case of a 62-year-old woman with a non-functional ACC of the left adrenal gland (T2 N0 M0, classified as stage II) who survived the disease for 28 years with 3 operations for metastases of the contralateral adrenal gland and 4 operations for metastases of the lung. We can now give an update of a further four years of survival and one additional operation. The case again emphasises the need for thorough radiographic examination and follow-up for the possibility of a repeated re-operation whenever possible to provide long-term survival over decades. PMID- 17465242 TI - Small cell cancer of the anal canal--case report of a rare tumor. AB - BACKGROUND: We report on a rare case of small cell cancer located at the anal canal. CASE REPORT: A 41-year old woman presented with locally advanced small cell anal cancer and simultaneous hepatic and pulmonal deposits. Due to metastatic disease, chemotherapy with etoposide and cisplatin was performed with mixed response after four cycles of chemotherapy. After application of two additional chemotherapy cycles, locally progressive disease occurred causing symptomatic bowel obstruction. Pelvic irradiation was started and, several days later, additional irradiation of cerebral metastases was initiated due to rapid progression of distant disease. Despite adequate local treatment the patient's condition further deteriorated and irradiation was stopped. The patient died 10 months after initial diagnosis due to rapid tumor progression. CONCLUSION: In patients with metastatic small cell anal cancer chemotherapy remains the mainstay of therapy. Radiotherapy exerts additional activity and remains a prime choice to gain local control and ameliorate symptoms. Careful histopathological examination, together with immunohistochemistry, is needed to determine the therapeutic strategy to be followed. PMID- 17465243 TI - Simultaneous proteomic and genomic analysis of primary Ta urothelial cell carcinomas for the prediction of tumor recurrence. AB - BACKGROUND: The prediction of tumor recurrence in patients with Ta urothelial cell carcinoma is inaccurate and new prognostic markers are desirable. MATERIALS AND METHODS: Surface-enhanced laser-desorption ionization time-of-flight mass spectrometry (SELDI-TOF MS) was performed on 33 primary Ta tumors (16 and 17 tumors were from patients with long and short recurrence-free periods, respectively) and data were compared to previously obtained mRNA expression profiles of 49 genes. RESULTS: The intensities of a protein peak at m/z 33331 varied most significantly between the two patient groups (p = 0.0048). This was comparable to survivin, whose mRNA expression differed most significantly (p = 0.0042) of the 49 genes. ROC analysis revealed an area under the curve for protein peak 33331 and survivin of 0.78 (95% CI, 0.62-0.94) and 0.79 (95% CI, 0.63-0.94), respectively. Protein peak 33331 and survivin identified 3 (17%) and 8 (47%) patients with a recurrence-free period of at least 4 years, respectively, without generating false-negatives. CONCLUSION: These findings indicate that SELDI-TOF MS and real-time Q-PCR analysis on the same tissue can result in the identification of markers with comparable differential expression. Such combined analyses may yield combinations of several markers that might improve disease prognosis. PMID- 17465244 TI - Maspin expression in adenocarcinoma of the ampulla of Vater: relation with clinicopathological parameters and apoptosis. AB - AIM: Maspin is a unique serine proteinase inhibitor which has tumor suppressor activity. The aim of the present study was to investigate the role of maspin in ampullary adenocarcinomas, its correlation with apoptosis and its value as a prognostic marker. PATIENTS AND METHODS: Twenty-three cases of ampulla of Vater adenocarcinoma were collected from archival material. For each sample, maspin, M30, p53 and Mib1 immunohistochemical reactivity were evaluated and results compared with clinicopathological parameters. RESULTS: A statistical relation was found between nuclear maspin and M30 (Spearman's Q = 0.46, p = 0.02), and p53 (Kruskal-Wallis = 0.03); a trend was found between nuclear maspin and pT (Kruskal Wallis = 0.09), and pM (Mann-Whitney = 0.08) and pN status (Fisher's mid-point test: p = 0.070). CONCLUSION: The present study evaluated the role of maspin in ampullary adenocarcinomas and for the first time demonstrated its association with apoptosis, tumor growth and metastasis. PMID- 17465245 TI - Temozolomide and thalidomide in the treatment of glioblastoma multiforme. AB - OBJECTIVE: The aim of this study was to assess efficacy and toxicity of temozolomide given alone or in combination with thalidomide, an anti-angiogenetic drug, in patients with newly diagnosed glioblastoma multiforme (GBM). PATIENTS AND METHODS: 46 patients with histologically proven GBM were eligible for inclusion. Twenty-three patients (15 males and 8 females) received temozolomide on a conventional schedule; 23 patients (12 males and 11 females) received temozolomide on the same schedule and thalidomide was dose-adjusted in each individual patient based on their tolerance. RESULTS: The median survival time was 12 months for temozolomide and 13 months for temozolomide + thalidomide. CONCLUSION: The administration of temozolomide in association with thalidomide after radiotherapy (RT) does not offer an advantage over temozolomide alone in adults with newly diagnosed GBM. The two therapeutic strategies produce similar results for survival, but the latter regimen shows a moderate increase in toxicity. PMID- 17465246 TI - Chromosomal aneusomy (chr 1, 11, 17) detected by fluorescence in situ hybridization may be a prognostic factor in breast cancer. AB - The relationship between clinicopathological findings and the long-term prognosis was investigated in 42 breast cancer patients in whom aneusomy was detected for chromosomes 1, 11 and 17. The frequencies of aneusomy of those chromosomes were 78.6%, 47.5% and 52.5%, respectively, and more than 90% of anomalies consisted of polysomy. The relationship between aneusomy and the clinicopathological findings showed a statistical correlation with a high histological grade in the case of polysomy of chromosome 17 compared with disomy, indicating a tendency for a high incidence of lymph node metastasis. Analysis of the survival data revealed that the prognosis was poor when there was polysomy of chromosomes 1 or 11. These results indicate the possibility that aneusomy of chromosomes 1, 11 and 17 can serve as prognostic factors of poor outcome in breast cancer patients. PMID- 17465247 TI - Primary rectal carcinoma in patients with stage IV resectable disease at diagnosis. AB - BACKGROUND: Rectal cancer is commonly diagnosed at a precocious stage, but for patients presenting at diagnosis with stage IV disease the best treatment is still undefined. The purpose of this study was to review the feasibility and outcome of multimodality treatment of rectal cancer patients metastatic at diagnosis. PATIENTS AND METHODS: From January 2000 to December 2005, 40 patients with histologically proven stage IV rectal adenocarcinoma (< 12 cm from the anal verge) were examined. Variables considered were age (under or over 65 years), tumour grade, presence of peritoneal carcinomatosis, type of surgery (palliative versus resection). RESULTS: The median age was 61 years (range, 32-83) and 27 were male and 13 female. Seventeen patients with unresectable or potentially resectable metastatic disease received neoadjuvant chemoradiotherapy (CHT-RT) with 5-fluorouracil (5FU) (plus oxaliplatin in 11 cases), radical surgery was performed in almost half of the cases; only in two patients were metastases also resected. If the patient is a candidate for radical surgical resection, the primary tumour should initially be treated as in a patient without metastatic disease and subsequently the primary tumour and metastases should be treated surgically. If the metastases are unresectable, the treatment of the primary lesion, according to the patient's symptoms, should be by palliative CHT. It is still not determined whether RT should be reserved for the symptomatic cases as an alternative to local surgery. In five patients treated with neoadjuvant CHT alone, radical local surgery was performed in two cases. Eighteen symptomatic patients were resected primarily; all of them received a postoperative CHT but only five of them also received postoperative RT. Nevertheless, after a multimodality treatment (neoadjuvant CHT +/- RT) 22.5% achieved a response rate (RR) (one complete remission (CR) and eight partial remission (PR)). Considering that all except two of the patients were locally radically resected and two of them also underwent metastases surgery, the overall RR was 17.5% (four CR and three PR). All of the CR were disease-free and alive after a median follow-up of 19.3 months. Age > or = 65 years had no impact on overall survival (OS), but the presence of peritoneal carcinosis in five patients showed a trend towards diminished survival, although it was not statistically significant (p = 0.08). CONCLUSION: The best treatment on diagnosis of metastatic rectal cancer is a multimodality CHT-RT approach. New prospective studies should evaluate non cross resistant regimens as additional therapy for those patients with a systemic residual disease after common CHT-RT. PMID- 17465248 TI - Risk factors for early age at breast cancer onset--the "COSA program" population based study. AB - BACKGROUND: Breast cancer is the most frequent cancer in women in western countries. A number of risk factors are now known, but the etiology of the disease is not fully understood. The aim of this study was to determine the effects of reproductive, anthropometric and environmental factors on cancer onset. PATIENTS AND METHODS: 934 women who developed a non-hereditary breast cancer were recruited from different hospitals in the Auvergne region (France) and completed a questionnaire. RESULTS: The use of oral contraceptives (OC) increased the risk of early cancer development (odds ratio = 1.84, 95% confidence interval = 1.38-2.44). The age at first OC use appeared to be a major factor since the risk decreased when OC use was after the age of 23 years (odds ratio = 0.52, 95% confidence interval = 0.34-0.79). A duration of breast-feeding greater than 26 weeks decreased the risk of early cancer development (odds ratio = 0.62, 95% confidence interval = 0.39-0.97). No overall association was found with anthropometric or lifestyle factors and early age at breast cancer onset. CONCLUSION: OC use, age at first OC use and lactation were significantly associated with an early age at breast cancer onset. Thus, a number of "risk factors" could be considered as "early onset risk factors". PMID- 17465249 TI - Actual chemotherapeutical possibilities in hormone-refractory prostate cancer (HRPC) patients. AB - Androgen deprivation therapy still remains the gold standard in the treatment of advanced prostate cancer. Unfortunately, patients with metastatic prostate cancer treated with androgen deprivation therapy frequently develop androgen-independent prostate cancer. Cytotoxic chemotherapy has not been used routinely and the current standard regimens have not demonstrated any significant alteration in the development of hormone-refractory disease. Recent phase III randomized clinical trials have suggested that docetaxel-based therapy, demonstrating a real increase of survival in treated patients, could represent the new standard treatment for metastatic patients. There is also promising activity of new drug combinations, such as taxanes plus vinca alkaloids, and of classic chemotherapeutic agents plus biological drugs. This review focuses on the current therapies for the treatment of HRPC. PMID- 17465250 TI - Cancer treatment-induced oral mucositis. AB - Oral mucositis is one of the main complications in non-surgical cancer treatments. It represents the major dose-limiting toxicity for some chemotherapeutic agents, for radiotherapy of the head and neck region and for some radiochemotherapy combined treatments. Many reviews and clinical studies have been published in order to define the best clinical protocol for prophylaxis or treatment of mucositis, but a consensus has not yet been obtained. This paper represents an updated review of prophylaxis and treatment of antineoplastic therapy-related mucositis using a MEDLINE search up to May 2006, in which more than 260 clinical studies have been found. They have been divided according to antineoplastic therapy (chemotherapy, radiotherapy, chemo-radiotherapy, high-dose chemotherapy). The prophylactic or therapeutic use of the analysed agents, the number of enrolled patients and the study design (randomized or not) were also specified for most studies. Accurate pre-treatment assessment of oral cavity hygiene, frequent review of symptoms during treatment, use of traditional mouthwashes to obtain mechanical cleaning of the oral cavity and administration of some agents like benzydamine, imidazole antibiotics, tryazolic antimycotics, povidone iodine, keratinocyte growth factor and vitamin E seem to reduce the intensity of mucositis. Physical approaches like cryotherapy, low energy Helium Neon laser or the use of modern radiotherapy techniques with the exclusion of the oral cavity from radiation fields have been shown to be efficacious in preventing mucositis onset. Nevertheless a consensus protocol of prophylaxis and treatment of oral mucositis has not yet been obtained. PMID- 17465251 TI - Evaluation of the quality of life between inpatients and outpatients receiving cancer chemotherapy in Japan. AB - A quality of life (QOL) assessment has become increasingly common in cancer clinical trials. Seventy-four consecutive patients treated for cancer between August 2005 and January 2006 at the Cancer Chemotherapy Center in the University of Occupational and Environmental Health, Japan, were examined. The 8-Short form health survey (SF-8) was utilized as a comprehensive scale and quality of life questionnaire for cancer patients treated with anticancer drugs (QOL-ACD) as disease specific scale for the QOL evaluation. The QOL for outpatients was investigated in comparison with that for inpatients. All questionnaires were collected and baseline questionnaires were filled in by 98.1% of the subjects. The physical comprehensive score (PCS) of SF-8 for the outpatients was higher than that for the inpatients. The physical condition of the outpatients was better than that of inpatients. There was no difference in the baseline scores of the QOL-ACD scales in daily activity, psychological condition, social attitude, and face scale of the analyzed domains between the two groups. Furthermore, a longitudinal study from admission to outpatient was carried out on 27 patients who were treated on an outpatient basis in our clinic. No difference in the baseline scores of the SF-8 and QOL-ACD scales were observed in any of the analyzed domains. These data suggest that the present QOL study has a sufficient feasibility for the outpatients evaluated in our study, and QOL of outpatients after discharge is equal to that of inpatients receiving cancer chemotherapy. PMID- 17465252 TI - Prediction of emergency hospitalization of outpatients receiving cancer chemotherapy. AB - The purpose of this study was to establish criteria to predict the need for emergency hospitalization of patients receiving chemotherapy, based on information at presentation. 158 consecutive patients treated for cancer at the Cancer Chemotherapy Center in the University of Occupational and Environmental Health were examined. The number of emergency hospitalization cases for outpatients undergoing cancer chemotherapy was 14 (8.9%) and including seven lung carcinomas, six hematological carcinomas, and one mediastinal tumor. The reason for emergency hospitalization in twelve (85.7%) of the cases was infection. No significant difference was observed between the cases with and without emergency hospitalization regarding age, gender, cancer type, previous treatment, objective of the chemotherapy, or line of chemotherapy. A significantly higher number of the emergency cases were associated with performance status 2, severe adverse events and comorbidity than with a performance status 0-1 where there were no or only mild adverse events and no comorbidity. Multiple logistic regression models indicated that severe adverse events and comorbidities were independent predictive factors for patients with emergency hospitalization. By combining selected clinical information for outpatients receiving cancer chemotherapy, the need for emergency hospitalization could be predicted. PMID- 17465253 TI - Tumor infiltrating dendritic cells predict treatment response to immmunotherapy in patients with metastatic renal cell carcinoma. AB - BACKGROUND: Although renal cell carcinoma (RCC) is considered to be an immunogenic tumor, the role of immunogenicity in this tumor for predicting treatment response has been little investigated. PATIENTS AND METHODS: Resected RCC specimens from 25 patients who received cytokine treatment for metastases were investigated using immunohistochemistry for CD83+ or S100+ dendritic cells (DCs), CD8+ T-cells, HLA-DR+ tumor cells, CD68+ tumor associated macrophages, microvascular density and vascular endotherial growth factor. RESULTS: Among the examined parameters, DCs status showed predictive value, that is, higher numbers of CD83+ or S100+ cells in tumors were associated with favorable treatment response. However, only higher CD83 status, which indicates mature and activated DCs, contributed to better survival (p = 0.0339). CONCLUSION: Increased tumor infiltration of mature DCs would be a predictor of treatment response and outcome in metastatic RCC patients, who receive immunotherapy. PMID- 17465254 TI - The role of imatinib mesylate in adjuvant therapy of extra-abdominal desmoid tumors. AB - BACKGROUND: Extra-abdominal desmoid tumors are rare neoplasms with variable biological behavior. The mainstay of treatment is surgery. Complementary treatment with tyrosine-kinase receptor inhibitor drugs, particularly imatinib mesylate, has been reported in the literature. The purpose of this study was to determine the possible presence of tyrosine-kinase receptors in extra-abdominal desmoid tumors as a marker for imatinib mesylate therapy. PATIENTS AND METHODS: From 1999 to 2004, immunohistochemical methods were carried-out in 14 patients with histologically confirmed extra-abdominal desmoid tumors to determine c-KIT positivity (existence of tyrosine-kinase receptors and PDGFRA and PDGFRB). RESULTS: All desmoid tumors were c-KIT negative, which demonstrates absence of tyrosine-kinase receptors. CONCLUSION: The histological c-KIT markup is an easy and reliable method that can detect whether a desmoid tumor is sensitive to additional treatment with a tyrosine-kinase receptor inhibitor. Molecular biological analysis for the identification of KIT and PDGFR mutation should be performed before imatinib mesylate is included in any treatment protocol. PMID- 17465255 TI - PSA measurement following prostatectomy: an unexpected error. AB - An unexpected and exceptional observation was made during the biological follow up of a patient who had undergone a radical prostatectomy for a prostate adenocarcinoma. A residual PSA level was first considered as a reliable marker of relapse. Certain limits are highlighted concerning the reliability of the PSA assay, which must be interpreted along with all the other prognostic information on the tumor and possibly verified using other biological techniques. PMID- 17465256 TI - Lower specific micronutrient intake in colorectal cancer patients with tumors presenting promoter hypermethylation in p16(INK4a), p4(ARF) and hMLH1. AB - BACKGROUND: The diversity of the Mediterranean diet and the heterogeneity of acquired epigenetic alterations in colorectal cancer (CRC) led us to examine the possible association between dietary factors and promoter hypermethylation in genes implicated in the pathogenesis of these neoplasms (p16(INK4a), p14(ARF), hMLH1) and the interaction with methylene tetrahydrofolate reductase (MTHFR) C677T polymorphism. PATIENTS AND METHODS: For the molecular study, 120 CRC patients were analyzed for hMLH1 promoter methylation status and MTHFR genotyping. Dietary patterns and molecular data on p16(INK4a) and p14(ARF) methylation were obtained from previous studies with this populations. RESULTS: Patients with methylation in p16(INK4a) consumed significantly less folate (p = 0.01), vitamin A (p = 0.01), vitamin B1 (p = 0.007), potassium (p = 0.03) and iron (p = 0.02) than controls. Patients with methylation in p14(ARF) or hMLH1 consumed significantly less vitamin A (p = 0.001 and p = 0.05, respectively). CONCLUSION: These results support that certain micronutrients protect against colorectal neoplasia and emphasize the importance of considering the different molecular forms of CRC as etiologically distinct diseases. PMID- 17465257 TI - Prognostic significance of HER-2, p53 and Bcl-2 in patients with epithelial ovarian cancer. AB - BACKGROUND: Several oncogenes and onco-suppressor genes have been implicated in epithelial ovarian carcinogenesis, but their clinical significance is not clear and conflicting data have been found in various studies. PATIENTS AND METHODS: The immunohistochemical expression of HER-2, p53 and Bcl-2 proteins was investigated in a cohort of 95 patients with advanced epithelial ovarian cancer (stages IIc-IV). These patients participated in a phase III randomized clinical trial and were treated either with paclitaxel/carboplatin, orpaclitaxel/carboplatin alternating with paclitaxel/cisplatin. RESULTS: Positive immunostaining for HER-2, p53 and Bcl-2 proteins was found in 18%, 70.5% and 69.5% of the cases, respectively. In multivariate analysis, older patients (< 63 vs. > or = 63 years, p < 0.001), worse grade (I-II vs. III, p = 0.04) and p53 expression (negative vs. positive, p = 0.002) were significant prognostic factors independently associated with survival. CONCLUSION: p53 status along with age and grade appear to be independent prognostic factors for survival in patients with epithelial ovarian cancer. PMID- 17465258 TI - Brain radiotherapy during treatment with anticonvulsant therapy as a trigger for toxic epidermal necrolysis. AB - Toxic epidermal necrolysis (TEN) is a severe mucocutaneous syndrome that can be occasionally caused by anticonvulsant drugs. In some cases, cranial irradiation may act as a precipitating factor. Thus, in cancer patients who suffer from brain metastases and are administered antiepileptic drugs for seizure prophylaxis, the risk of developing TEN after receiving palliative brain radiotherapy cannot be ignored. We is reported. The case of a young patient with non-small cell lung cancer (NSCLC) treated with prophylactic phenobarbital who developed TEN within a few days of completing cranial radiotherapy for brain metastases is reported. To minimize the risk of TEN in patients undergoing brain radiotherapy, prophylactic anticonvulsant therapy is recommended only after an accurate measurement of the true benefits. Alternatively, discontinuation of antiepileptic treatment before the initiation of brain radiotherapy, or the use of anticonvulsants associated with a lower risk of developing cutaneous reactions might be considered. PMID- 17465259 TI - Prognostic significance of ezrin expression in pleomorphic malignant fibrous histiocytoma. AB - BACKGROUND: Newly defined pleomorphic malignant fibrous histiocytoma represents a pleomorphic sarcoma which shows no definable line of differentiation. In this study, the survival and validity of known prognostic factors for patients with pleomorphic MFH, with the possibility of ezrin expression as a new prognostic factor were investigated. PATIENTS AND METHODS: Forty-seven patients without metastasis at presentation were analyzed. Immunohistochemical analysis with anti ezrin antibody was performed using tissue microarray. RESULTS: The overall, metastasis-free, and local recurrence-free survival rates were 59%, 57%, 52% at 5 years, respectively. Uni- and multivariate analyses revealed that truncal location and ezrin positivity had negative effects on overall and metastasis-free survival. The independent factors of local recurrence-free survival were narrow surgical margin, truncal location and female gender. CONCLUSION: Ezrin expression was an independent predictor of distant metastasis in pleomorphic MFH. Thus, ezrin seems to provide additional prognostic information and may be a novel target for the development of new adjuvant therapies. PMID- 17465260 TI - Pharmacokinetics and toxicity of an early single intravesical instillation of gemcitabine after endoscopic resection of superficial bladder cancer. AB - BACKGROUND: The tolerability and plasma absorption of gemcitabine administered at 40 mg/ml after small and extensive endoscopic transurethral resection of bladder tumors (TURB) were evaluated. PATIENTS AND METHODS: Nine patients with a history of recurrent superficial bladder cancer were eligible for a single immediate, post TURB, intravesical instillation of gemcitabine. The endoscopic resection was small in 5 patients and extensive in 4. The drug was administered at 40 mg/ml concentration (2000 mg in 50 ml saline) and held in the bladder for 1 hour. Plasma concentrations of gemcitabine and its metabolite (2',2' difluorodeoxyuridine) were determined with a validated HPLC assay. The blood count and chemistry were performed one day and one week postoperatively. RESULTS: Toxicity was comparable for patients who underwent small or large TURB. The most significant side-effects were grade 2 vomiting and a transient grade 2 leukopenia after small and large TURB respectively. Mean maximum gemcitabine concentrations were 1.47 microg/ml in small TURB and 2.8 microg/ml in large TURB. The highest peak concentration of 4.26 microg/ml was found after extended bladder resection. CONCLUSION: A single, immediate postoperative, intravesical instillation of gemcitabine at high concentration is feasible with acceptable toxicity, and it may be considered as an option taking into account patient performance status, tumor characteristics and TURB extension. PMID- 17465261 TI - Usefulness of homocysteine as a cancer marker: total thiol compounds and folate levels in untreated lung cancer patients. AB - BACKGROUND: Apart from being a risk factor for atherosclerotic cardiovascular diseases, the latest research suggests homocysteine as a marker for cancer. We aimed to explore the clinical utility of plasma homocysteine levels as a marker in lung cancer. PATIENTS AND METHODS: Changes in serum total thiols and folate levels were investigated in newly diagnosed untreated lung cancer patients (n = 37) and compared with healthy controls (n = 26). Fluorometric HPLC methods were used for the determination of thiols. Other parameters were determined with commercial diagnostic kits. RESULTS: Increased total homocysteine (t-Hcy), decreased total glutathione (t-GSH) and folate levels were observed in lung cancer patients compared with healthy controls. Total levels of thiols and folate did not show any significant difference between SCLC and NSCLC patients. However, there were significantly higher t-Hcy, lower t-GSH and folate levels in the advanced-stage group compared with controls. Prevalence of hyperhomocysteinemia was 65% in lung cancer patients when 12 micromol/l were taken as a cut-off value for t-Hcy levels. CONCLUSION: Homocysteine is suggested as a marker for several types of cancer, but our result did not support this hypothesis for lung cancer. Although higher homocysteine levels were observed in the present study, further investigation in the larger cancer population would clarify the importance of homocysteine as a cancer marker. PMID- 17465262 TI - Survival improvement of young patients, aged 16-23, with Hodgkin lymphoma (HL) during the last three decades. AB - The prognostic factors, treatments and outcomes of 55 young adults (16-23 years old) with Hodgkin lymphoma (HL) treated in the Second Department of Internal Medicine Propaedeutic, Medical Oncology Unit, Athens University, over the past 25 years, are reviewed. Patients were treated with the chemotherapy regimens available at each time period which were MOPP (Group A; 1978-1987), MOPP/ABVD (Group B; 1988-1993) and BEACOPP or ABVD (Group C; 1994-2003). The eligible patients, received radiotherapy (RT) according to treatment consensus. Additionally, the patients were retrospectively divided according to risk factors (abnormal erythrocyte sedimentation rate (ESR), bulky mediastinal disease, > 3 involved nodes and extranodal involvement) into low [stage I/II; five patients (9%)], intermediate [stage III with adverse prognostic factors; 18 patients (33%)] and high risk categories [stages IIB bulky and III/IV; 32 patients (58%)]. A total of 21 (38%) patients experienced relapse (three intermediate and 19 high risk). The 5-year survival and the 5-year event free survival (EFS) figures were Group A: 65% and 53%, Group B: 80% and 65%, Group C: 100% and 88.5%, respectively, the improvements between Group B and C were statistically significant (p = 0.04 and p = 0.005, respectively) among the three time periods. The overall survival (OS) and EFS differed significantly between intermediate and high risk categories (OS: p = 0.04, EFS: p = 0.005). The sequential use of RT did not influence OS and EFS but there was a trend of improvement with RT in the later periods. Survival of young patients with HL is significantly improving most probably due to improved chemotherapy treatment and understanding of the risk factors. Current controversial issues surrounding this disease, including the role of radiotherapy, positron emission tomography (PET), bone marrow biopsy and stem cell transplantation are discussed. PMID- 17465263 TI - Shorter CAG repeat in the AR gene is associated with atypical hyperplasia and breast carcinoma. AB - BACKGROUND: Previous reports into the role of [CAG]n repeat lengths in the androgen receptor (AR) gene indicate that these may play an important part in the development and progression of breast cancer, however, knowledge regarding benign breast lesions is limited. PATIENTS AND METHODS: PCR-based GeneScan analysis was used to investigate the [CAG]n repeat length at exon 1 of the AR gene in 59 benign breast lesions (27 fibroadenomas, 18 atypical hyperplasias, and 14 hyperplasias without atypia) and 54 ductal breast carcinomas. Seventy-two cancer free women were used as a control group. In addition, [CAG]n repeats were evaluated for the presence of loss of heterozygosity (LOH) and microsatellite instability (MSI) in a subset of these samples (27 fibroadenomas, 14 hyperplasias without atypia and 22 breast carcinomas). RESULTS: Shorter [CAG]n repeat lengths were strongly correlated with atypical hyperplasias (p = 0.0209) and carcinomas (p < 0.0001). LOH was found in 1/12 and 4/20 informative cases of hyperplasias without atypia and breast carcinomas, respectively. Three patients with breast carcinoma who had previously presented atypical hyperplasia showed a reduction in the [CAG]n repeat length in their carcinomas. CONCLUSION: Short [CAG]n repeat length (< or = 20) polymorphisms are strongly associated with breast carcinomas and atypical hyperplasias. Although non-significant, a subgroup of patients with breast carcinoma and genotype SS showed an association with parameters of worse outcome. PMID- 17465264 TI - Circulating cell-free mRNA in plasma as a tumor marker for patients with primary and recurrent gastric cancer. AB - BACKGROUND: The diagnostic value of circulating mRNA for the early detection of primary and recurrent gastric cancer was evaluated. PATIENTS AND METHODS: Circulating hTERT and MUC1 mRNA were amplified in the plasma from 52 gastric cancer patients (40 preoperative and 12 postoperative patients) and 20 healthy controls. The results were compared with those of a circulating cancer cell assay and methylation-specific polymerase chain reaction assay. RESULTS: Cell-free mRNA of the analyzed genes was detected in 6 (15%) preoperative gastric cancer patients (hTERT: 3 and MUC1: 4 patients) and 2 follow-up patients. These mRNAs were not detected in the plasma from healthy volunteers. There was no correlation between the results of the cell-free mRNA and the other assays. CONCLUSION: Detection of circulating cell-free mRNA might serve as a new complementary tumor marker for gastric cancer. PMID- 17465265 TI - Prognostic factors and treatment-related outcome in patients with uterine papillary serous carcinoma. AB - A retrospective analysis was performed in order to evaluate prognostic factors and treatment-related outcome in patients with uterine papillary serous carcinoma (UPSC). PATIENTS AND METHODS: Between 1989 and 2003 forty-five patients with UPSC were treated at the McGill University Health Centre (MUHC), and therefore were included in the analysis. Age, race, history, tumor stage and grade, invasion and metastasis, treatment and outcome for each patient were evaluated. RESULTS: According to FIGO classification, Stage I, II, III, and IV tumors were identified in 36%, 9%, 44%, and 11% of the patients, respectively. At the time of analysis, 37.8% of patients (17/45) were dead due to disease, with a mean survival of 22 months (SD +/- 7.5 months). Cause-specific survival for the entire group was 69%, 66%, and 58% at 2, 3, and 5 years, respectively. With respect to disease stage, 5 year cause-specific survival for stage I, II, III, and IV was 100%, 75%, 43%, and 0%, respectively. Univariate analysis comparing cause-specific survival curves demonstrated a statistically significant difference for disease stage (p < 0.0001) and depth of myometrial invasion (p = 0.008). However, in multivariate analysis, only disease stage had a significant impact on cause-specific survival (p < 0.01). CONCLUSION: Disease stage is the only independent significant prognostic factor regarding cause-specific survival in patients with UPSC. PMID- 17465266 TI - Prevalence of hepatitis C virus (HCV) genotypes and increase of type 4 in central Italy: an update and report of a new method of HCV genotyping. AB - Hepatitis C virus (HCV) genotyping is very useful for identifying the patients (type 1 and 4) that need more aggressive management. In recent years, genotype 4 has shown spread in different parts of Europe. The aim of this study was to update on the prevalence of HCV genotypes of 288 patients in Central Italy, to analyze the possible increase of genotype 4 and to evaluate a new simple genotyping method. A line-probe assay (LiPA, Bayer) was used based on the reverse hybridization of HCV genome fragments previously amplified and biotinylated by a polymerase chain reaction (PCR) assay, COBAS System Amplicor HCV monitor version 2.0 (Roche) or previously amplified by COBAS Ampliprep/TaqMan HCV test (Roche). This last method uses non-biotin-labeled primers, therefore we added for each sample 10 microl of amplified HCV products, 10 microl of denaturation solution and 10 microl of biotinylated-nested primers (Bayer) to utilize the genotyping procedure previously used. The results showed that the prevalence of type 1, 2 and 3 (482, 34.6 and 10.5%, respectively) as well as the prevalent subtypes, 1b and 2a/2c (30.7 and 27.2%, respectively) were similar to previous data. Type 1 and 2 were statistically associated with an older group of patients when compared with type 3 and 4 (p < 0.001). Type 3 and 4 showed a significant prevalence of male patients compared to type 1 and in particular to type 2 (p < 0.014). The prevalence of type 4 was 5.6% in 2004, 6.1% in 2005 and 9.9% from January to July 2006. Type 4 showed an increase of male prevalence over a 3-year period (p < 0.001). In conclusion, subtype 1b and 2a/2c showed a very similar prevalence, age and gender distribution in Central Italy. The type 4 patient group was analyzed because an increase of this genotype (1.8 times) was detected. PMID- 17465267 TI - Prognostic impact of haemoglobin levels in breast cancer. AB - BACKGROUND: Many patients with solid tumours suffer from anaemia, as a consequence of the disease itself or its treatment. Anaemia affects the quality of life and can have a negative impact on overall survival. The aim of the current study was to analyse the impact of haemoglobin levels on the prognosis of patients with primary breast cancer. PATIENTS AND METHODS: Retrospective data from 249 women treated for operable breast tumours were included in our study. Haemoglobin (Hb) levels independently of anticancer therapy were compared for the prognostic influence on disease-free and overall survival. RESULTS: A significant correlation between higher minimum Hb level during chemotherapy and the disease free and overall survival was found. Pre-treatment haemoglobin levels had no prognostic influence on the disease-free and overall survival. CONCLUSION: The present data showed that anaemia during adjuvant chemotherapy to be a negative prognostic indicator for survival of patients with breast cancer. PMID- 17465268 TI - Effects of manganase superoxide dismutase Ala-9Val polymorphism on prostate cancer: a case-control study. AB - BACKGROUND: Manganese superoxide dismutase (MnSOD) is a major enzyme that is responsible for the detoxification of reactive oxygen species in the mitochondria. Mitochondrial DNA damage may contribute to carcinogenesis as an important risk factor. The aim of this study was to investigate the relationship between prostate cancer and MnSOD Ala-9Val polymorphism in Turkish men with prostate cancer. PATIENTS AND METHODS: Fifty patients with prostate cancer and 50 healthy controls were included in this study. Gene polymorphism was determined using a PCR-RFLP method. RESULTS: The Ala/Ala genotype and the Ala allele were found at statistically higher frequencies in patients with prostate cancer as compared to controls (p < 0.05). The patients suffering from prostate cancer were divided into two groups according to Gleason score: aggressive prostate cancer and non-aggressive prostate cancer. It was observed that carrying the Ala/Ala genotype or the Ala allele resulted in an insignificant increase in the frequency of aggressive prostate cancer compared to nonaggressive prostate cancer. It was concluded that MnSOD Ala allele might be the cause of prostate cancer risk among alcohol users. CONCLUSION: The results of our study of Turkish prostate cancer patients suggest that mutation of the MnSOD gene may be an important risk factor for prostate cancer. PMID- 17465269 TI - [Electrophysiological investigation of the texture discrimination mechanisms]. AB - In electrophysiological and psychophysical experiments, we investigated mechanisms of the visual system underlying local and global texture processing. Textures included rectangular matrixes composed of Gabor patches (sine wave grating windowed by a Gaussian envelope). Orientation of each grating varied from 0 to 165 degrees with the step of 15 degrees. Matrixes differed by the amount of Gabor patches with vertical or horizontal orientation. The observers' task was to discriminate the dominant orientation. The advantage of such stimuli involved a possibility to calculate global statistics of the textures, which we considered as the difference between whole amount of vertical and horizontal orientations in the stimulus irrespective of their location. The local statistics was calculated as relative amount of spatially organized nearby gratings (i. e. collinear contours). The subjects' accuracy was low in discriminating less organized textures and gradually improved with the amount of vertically of horizontally oriented Gabor patches, while the reaction time decreased. Visual evoked potentials (VEPs) recorded from occipital lobes revealed different dependencies of their components' magnitude on the amount of equally oriented gratings. Amplitude of the late positive component P3 with latency 400 ms directly depended on the texture discriminability, and N2 wave with latency 180 ms had an S-like dependence. Opposite to that, the magnitude of P2 wave with latency 260 ms was maximal in response to less organized textures and gradually decreased with the amount of equally oriented gratings. The dependencies received were compared with the textures' statistics. Data analysis allowed us to suppose that, in the conditions of our experimental paradigm, two mechanisms were involved in discrimination of the textures--the local and the global processing. We believe that by recording VEPs one can separately investigate activity of these two processes. PMID- 17465270 TI - [Individual alpha activity of electroencephalogram and nonverbal creativity]. AB - The main objective of present correlational investigation was to clarify relationships between nonverbal creativity indices and individual electroencephalogram alpha activity indices: individual alpha peak frequency, alpha band width, magnitude of alpha desynchronization and alpha spindle indices such as duration, amplitude, variability and skewness. The EEG was recorded in 98 healthy male right-handed subjects. Scores of nonverbal creativity (i. e., fluency, originality, and flexibility) were assessed using the Torrance test of nonverbal performance. The study showed that fluency in creative performance was associated with individual alpha peak frequency and alpha spindles duration, whereas originality and plasticity--with individual alpha band width and spindle amplitude variability. The findings also show that both highest and lowest individual alpha peak frequency indices are associated with enhanced scores of originality. It is suggested that individual alpha activity indices could be presented as individual predictors of fluency, plasticity and originality of nonverbal creativity. PMID- 17465271 TI - [Physiological effects of using the low intensity strength training without relaxation in single-joint and multi-joint movements]. AB - The effects of classical strength training (CT) and low intensity strength training without relaxation (TwR) upon size, strength and fatigability of leg muscles in men were compared. A 8-10-week strength training led to an increase of size and maximal voluntary contraction of trained muscles. After the CT, the increment of strength was higher; on the other hand, strength increments related to total work performed increased after the TwR noticeably higher than after the CT. Two training programs influenced the size of total muscle and of muscle fibers (MF) differently: the volume of m. quadriceps femoris increased more after the CT than after the TwR. The CT induced a significant increase of cross sectional area (CSA) of fast MF, and the TwR induced an increase of CSA of slow MF. Resistance to fatigue after the TwR was higher than after the. The effects of TwR were more pronounced in single-joint movements training than in multi-joint movement. PMID- 17465272 TI - [Transport function of lymph nodes in body antiorthostatic posture]. AB - The spontaneous contractile activity of isolated lymph nodes and the lymph flow from intestine lymphatic vessel in antiorthostatic posture of rats with an inclination angle of 30 degrees during 7-14 days, was decreased. Contractions of the rat lymph nodes in response to actions of adrenaline, acetylcholine and histamine were diminished. There are changes of biochemical components of lymph and blood plasma with simultaneous decrease of the blood plasma volume. It is concluded that the lymphatic system on antiorthostatic posture plays the compensatory role with the purpose of stabilization of homeostasis in the brain. PMID- 17465273 TI - [Coagulation hemostasis and fibrinolytic potential of blood in conditions of chronic stress and terahertz therapy]. AB - The effects of electromagnetic rays of maximum high frequencies of radiation molecular spectrum and absorption of nitrogen oxide 150, 176-150, 664 GHz on blood coagulation properties of white laboratory rats subjected to chronic immobilization stress have been studied. It is shown that preventive course of electromagnetic irradiation with terahertz range at the frequencies of molecular spectrum of radiation and absorption of nitrogen oxide 150, 176-150, 664 GHz warns about development of stress disturbances of coagulation component of the hemostasis system and fibrinolysis in animals. PMID- 17465274 TI - [Effect of corticoliberin fragment CRF4-6 on blood glucose level and body temperature of rats]. AB - The effects of tripeptide corticoliberin fragment CRF4-6 (Pro-Pro-Ile) on blood glucose level and the rat body temperatire were investigated. Intracerebroventricularly injected CRF4-6 (6, 30, 150 nmol/head) causes a dosedependent hyperglycemia and hyperthermia in anaesthetized animals. Corticotropin releasing factor antagonist alpha-helical CRF4-6 (6.5 nmol/head) abolishes the influence of tripeptide CRF4-6 (6 nmol/head) on blood glucose level and body temperature of rats. Bilateral adrenalectomy has no effect on tripeptide induced hyperglycemia and hyperthemia. This result indicates that hyperglycemic and hyperthermal effects of tripeptide occur independently of adrenal gland catecholamines. In addition, non-pituitary corticoliberin receptors are involved in CRF4-6 influences on blood glucose level and body temperature. PMID- 17465275 TI - [Corticosteroid hormones in pathogenetic heterogenety of post-stress depression]. AB - The role of corticosteroid hormones in post-stress depression progress and their usage for correction of this psychopathology was studied in active and passive Wistar rats. It was found that only passive individuals had sensitivity to corticosteroids and, among them, only adrenal gland hormones lead to corrections of post-stress depression. PMID- 17465276 TI - [Development of protective functions of the pig digestive tract in ontogenesis]. AB - It has been found that the state of protective functions of the digestive tract change during postnatal period. The biosynthesis and secretion of glycoproteins of mucus layer which is the main protective structure of digestive tract depend on the hormonal background and the nutrition conditions. The biosynthesis stages and secretion glycoproteins result in sexual maturity and in conditions of definitive nutrition. Opposite to that, the antiradical activity of adherent mucous layer decreases with age. The allocation of the studied components of adherent mucous layer in the stomach and in the intestine is determined. PMID- 17465277 TI - [On the mechanism of gastroduodenal motility inhibition under psychogenic stress in rabbits]. AB - In experiments on unanaesthetized rabbits, myoelectric activity (contractile activity index) in antral and pyloric parts of the stomach and in two sites of proximal duodenum was studied under stress induced by fastening rabbit to a table in supine position. The stressor impact induced inhibition of contractile activity in antrum and pylorus. The duodenal contractile activity after initial complete suppression overshot its initial level. Blockade of beta1/beta2 adrenoceptor with propranolol and blockade of alpha2-adrenoceptor with yohimbine did not influence qualitatively the pattern of the stressor responses of antrum and pylorus, and of the postpyloric part of duodenum. In conditions of unselective blockade of alpha-adrenoceptor with dihydroergotoxin there was no initial complete inhibition of duodenal contractile activity, and its strengthening was more expressed than in the control experiments. The received data indicate that the stressor inhibition of antral and pyloric contractile activity possibly results from activation of non-adrenergic inhibitory neurons of the enteric nervous system. The initial short-term suppression of duodenal motility resulted from its "adrenergic" inhibition which can also be a factor limiting the manifestation of stimulating effect of the humoral agent on the duodenal motility. In the period after release of the animal, index of antral and pyloric contractile activity did not significantly differ from its initial level; after beta1/beta2-adrenoceptor blockade in antral and after alpha2-adrenoceptor blockade or nonselective alpha-blockade in antral and pyloric parts of the stomach, there was decrease of contractile activity compared with its initial level; after alpha2- or beta1/beta2-adrenoceptor blockade there was no poststressor exceeding of the initial level of the duodenal contractile activity, observed in the control experiments. PMID- 17465278 TI - [Light modes and ovulation function in rats in ontogenesis]. AB - Enhancement of illumination was shown to lead to occurrence of early and more obvious age-dependent changes of ovulation cycle in rats as compared with the same changes in animals staying in conditions of light deprivation or standard illumination. The effect of 24-hour illumination during a year induced more obvious and still earlier changes of the estrus cycle that the effect of natural illumination in the North-West of Russia with its peculiar yearly photo periodicity (short light day in the autumn-winter period and "white nights" in the spring-summer period). PMID- 17465279 TI - Plant lipid transfer proteins (LTPS): biochemical aspect in panallergen- structural and functional features, and allergenicity. AB - Lipid transfer proteins (LTPs) are highly conserved and widely distributed throughout the plant kingdom. Several members of LTP family have been identified as relevant allergens in food and pollens. Because of their high resistance to heat treatments and enzymatic digestion, these proteins are allergenic candidates for oral route sensitisation. This review presents biochemical features, allergenicities and cross reactivities of fruit, cereal and pollen LTPs. PMID- 17465280 TI - Role of tropomyosin as a cross-reacting allergen in sensitization to cockroach in patients from Martinique (French Caribbean island) with a respiratory allergy to mite and a food allergy to crab and shrimp. AB - BACKGROUND: Tropomyosin has been described as cross-reacting allergen between mite, cockroach and shrimp. METHODS: In 13 patients with asthma and/or rhinitis sensitized to mite and/or German cockroach and presenting urticaria, oral allergy syndrome or angio-edema upon eating shrimp and/or crab, we measured specific IgE to mite, cockroach, crab and shrimp tropomyosin. RESULTS: Ten patients had specific IgE to tropomyosin from mite, 8 from shrimp, 6 from crab and 5 from cockroach. AST inhibition tests indicated that mite allergen is a primary sensitizer and is cross-reacting with shrimp, crab and cockroach allergens. CONCLUSION: Tropomyosin could be the cross-reacting allergen relevant for clinical symptoms to mite, cockroach, shrimp and crab. PMID- 17465281 TI - Respiratory allergy in agriculture. AB - Allergic respiratory diseases in farmers may be caused by exposures to many organic substances. Potentially inhalable particulate material of biologic origin are referred to collectively as organic dust, whose composition includes also molds and other microorganisms. Organic dust may evoke immuno-allergic reactions and cause rhinitis, asthma and extrinsic allergic alveolitis. The agricultural work environment represents a risk factor for these diseases, whose occupational origins are often overlooked by clinicians. Prevalence studies of respiratory allergic diseases among agricultural workers are advocated for the development of prevention strategies. PMID- 17465282 TI - Sublingual immunotherapy: from safety to mechanism of action. AB - Allergen specific immunotherapy is an important option for the treatment of respiratory allergy and its clinical efficacy has been clearly demonstrated by several studies. However, the injective route of administration and the possibility of severe side effects has limited its use in children and led to the introduction of new forms of administration. Sublingual immunotherapy (SLIT) has proven to be an effective and safe treatment for respiratory allergy. However, its mechanism of action is still debated. Pharmacokinetic studies showed that, differently from nasal mucosa, allergen extracts administered by SLIT are not immediately adsorbed but are long retained before being drained to local lymph nodes. This difference may be responsible of the absence of severe side effects and instead of short-lasting local symptoms. Studies by biopsies of the oral mucosa should greatly help in defining the presence and the role of cells involved in the mechanisms of oral tolerance. PMID- 17465283 TI - External fixation of tibial pilon fractures and fracture healing. AB - Distal tibial fractures are rare and difficult to treat because the bones are subcutaneous. External fixation is commonly used, but the method often results in delayed union. The aim of the present study was to find out the factors that affect fracture union in tibial pilon fractures. For this purpose, prospective data collection of tibial pilon fractures was carried out in 1998-2004, resulting in 159 fractures, of which 83 were treated with external fixation. Additionally, 23 open tibial fractures with significant > 3 cm bone defect that were treated with a staged method in 2000-2004 were retrospectively evaluated. The specific questions to be answered were: What are the risk factors for delayed union associated with two-ring hybrid external fixation? Does human recombinant BMP-7 accelerate healing? What is the role of temporary ankle-spanning external fixation? What is the healing potential of distal tibial bone loss treated with a staged method using antibiotic beads and subsequent autogenous cancellous grafting compared to other locations of the tibia? The following risk factors for delayed healing after external fixation were identified: post-reduction fracture gap of >3 mm and fixation of the associated fibula fracture. Fracture displacement could be better controlled with initial temporary external fixation than with early definitive fixation, but it had no significant effect on healing time, functional outcome or complication rate. Osteoinduction with rhBMP-7 was found to accelerate fracture healing and to shorten the sick leave. A staged method using antibiotic beads and subsequent autogenous cancellous grafting proved to be effective in the treatment of tibial bone loss. Healing potential of the bone loss in distal tibia was at least equally good as in other locations of the tibia. PMID- 17465284 TI - Immediate neonatal outcomes after elective induction of labor. AB - OBJECTIVE: To examine immediate neonatal outcomes associated with elective labor induction. STUDY DESIGN: Labor inductions occurring at > or = 38 weeks' gestation were examined during a 6-month period at 2 community hospitals. Medical records were reviewed by trained abstractors to determine the reason for induction (elective vs. medical) and maternal characteristics. The need for newborn resuscitation (1-minute Apgar score < 4) was the primary end point. Multiple logistic regression was used to calculate odds ratios (ORs) and 95% confidence intervals (CIs). RESULTS: Of the 364 inductions, 54.9% were elective. The odds of a 1-minute Apgar score being < or = 3 were significantly greater when labor was induced for elective reasons than for medical reasons (OR 5.5, 95% CI 1.1-27.9) or was spontaneous (OR 6.5, 95% CI 2.4-17.8), after controlling for mother's age, race and route of delivery. Elective induction was not associated with feal intolerance to labor, a low 5-minute Apgar score or need for admission to a special care nursery. CONCLUSION: An elective abortion induction is an independent risk factor for delivery of an infant requiring immediate attention. PMID- 17465285 TI - Diagnosis of adenomyosis: a review. AB - This article reviews the various preoperative diagnostic methods for uterine adenomyosis. MEDLINE and PubMed were searched using the keywords adenomyosis and adenomyosis diagnosis. Reviews, case-controlled studies and reports published from 1949 through March 2005 and written, or at least abstracted, in English were analyzed. Transvaginal sonography is superior to the transabdominal route, but its diagnostic sensitivity is still suboptimal, ranging from 50% to 87%. Magnetic resonance imaging is most effective for both diffuse and focal adenomyosis, with sensitivity and specificity that are comparable to or even better than those of sonography as it depicts contrasts between low-intensity lesions and surrounding tissue. Computed tomography has poor diagnostic value due to similar images portrayed by foci and normal myometrium. Myometrial biopsy of the posterior uterine wall, as pursued by some authorities in recent years, is superior to sonography, but its routine use is not recommended. Although various methods were added to the clinician's armamentarium over the last 2 decades, the preoperative diagnosis of adenomyosis remains challenging. PMID- 17465286 TI - Risk-adjusted cesarean rates: what risk factors for cesarean delivery are important to practicing obstetricians? AB - OBJECTIVE: To determine which primary cesarean delivery risk factors are important to practicing physicians. STUDY DESIGN: A sample of current members of the American College of Obstetrician Gynecologists were surveyed about the risk factors for primary cesarean delivery that they thought were most important. Data on demographic and practice characteristics were also collected. Sample size was determined by theoretical saturation. RESULTS: Theoretical saturation was reach at 60 responses. A total of 290 surveys were returned. The 10 most common factors listed by physicians as preexisting patient risk factors for primary cesarean delivery were, in descending order of importance, medical problems, maternal obesity, macrosomic infant, malpresentation, multiple gestation, maternal age, Bishop score, patient's fear, preterm labor, and postdate pregnancy. Six of the 10 factors listed by respondents are in previously published risk-adjustment models. CONCLUSION: Our study suggested that the addition of obesity and birth weight to previously published risk-adjustment models should improve representation of practicing obstetricians' views. PMID- 17465287 TI - Effect of test order on sensitivity in vulvodynia. AB - OBJECTIVE: To assess the effect of order of sensitivity testing at the vulva and thumb on the sensitivity determined at the second site tested among women with and without vulvodynia. STUDY DESIGN: We evaluated the stability of sensitivity measurements to pressure at the vulva and thumb when the order of testing was randomized to vulva first vs. thumb first; we repeated testing 1 week later in the opposite order. RESULTS: Stability of results over time and the influence of the order of testing were determined among 13 women with vulvodynia and 20 asymptomatic control women. We found a strong correlation between results compared between the first and second visits as well as no order effect. CONCLUSION: The order of testing at vulvar and peripheral sites has little impact on the results of pressure-responsive sensitivity testing among women with and without vulvodynia. PMID- 17465288 TI - Expression of CD44s, vascular endothelial growth factor, matrix metalloproteinase 2 and Ki-67 in peritoneal, rectovaginal and ovarian endometriosis. AB - OBJECTIVE: To understand the pathogenetic mechanisms of endometriosis by examining the expression of adhesion molecules (CD44s), angiogenic factor (VEGF) and matrix protease and to perform Ki-67 labeling for evaluation of proliferative activity. STUDY DESIGN: Twenty-nine peritoneal endometriosis lesions (9 red, 12 black and 8 white), 11 rectovaginal and 22 ovarian were obtained. Immunohistochemical staining was performed with antibodies for CD44, VEGF, MMP-2 and Ki-67. RESULTS: CD44s were expressed mainly in stroma and showed higher expression in glandular epithelium of peritoneal endometriosis than in rectovaginal and ovarian endometriosis. The stroma in red and white lesions showed higher MMP-2 expression than in black lesions. The stromal cells in rectovaginal endometriosis showed significantly lower expression of Ki-67 (p = 0.002) than in peritoneal and ovarian endometriosis. When endometriosis was analyzed according to the revised American Fertility Society classification, Ki 67 expression was high in glandular epithelium in stages I and II (p = 0.025), whereas MMP-2 expression in stromal cells was significantly high (p < 0.001) in stages III and IV. CONCLUSION: CD44, VEGF and MMP-2 were consistently expressed in endometriotic epithelial and stromal cells. White lesions of peritoneal endometriosis should not be regarded as an inactive state, and MMP-2 in stromal cells may be responsible for the progression of endometriosis. The macroscopic appearance of endometriotic lesions should not be used as a criterion to define the degree of activity. PMID- 17465289 TI - Indicated vs. routine prenatal urine chemical reagent strip testing. AB - OBJECTIVE: To determine if urinary tract infection, high blood pressure and gestational diabetes mellitus (GDM) are underdiagnosed when prenatal urine testing is done on a clinically indicated vs. routine basis. STUDY DESIGN: Prenatal and delivery records of 2,981 subjects were reviewed in a predominantly Hispanic and medically underserved population. Patients prior to August 2002 received routine urine screening. After August 2002, women were tested only if preestablished criteria were present. RESULTS: The number of patients diagnosed with high blood pressure and urinary tract infection was equivalent in the 2 groups. Despite unchanged screening for GDM between groups, the incidence of GDM declined from 9.3% to 4.2%. CONCLUSION: GDM, high blood pressure and urinary tract infection will not be underdiagnosed if prenatal urine testing is done on an indicated basis. It is safe to discontinue routine urine screening when a regimen of initial urine cultures, GDM screening at 24-28 weeks, indicated urine chemical reagent strip testing and routine blood pressure determination is used. Criteria for indicated urine testing should be clearly defined and consistently followed by all staff. PMID- 17465290 TI - Pelvic actinomycosis with retained intrauterine fetal bone: a case report. AB - BACKGROUND: Pelvic actinomycosis is a rare infection that can manifest as pelvic inflammatory disease and in severe cases can cause extensive fibrosis. Most cases are associated with long-standing use of an intrauterine device (IUD). CASE: A 30 year-old woman presented with abdominal pain, fever and a pelvic mass. She underwent removal of an intrauterine foreign body, surgical drainage of a tuboovarian abscess and intravenous antibiotic therapy. Pathology studies revealed that the foreign body consisted of bone tissue, and the agent of infection was identified as Actinomyces israelii. CONCLUSION: Pelvic actinomycosis, although usually occurring in women using an IUD, may result from retained intrauterine fetal bone through a similar pathogenesis. PMID- 17465291 TI - Benign lipoblastomalike tumor of the vulva: report of a case affecting a young patient. AB - BACKGROUND: Mesenchymal vulvar tumors are rare, and a benign lipoblastomalike tumor may be confused with malignancy. CASE: A 15-year-old patient consulted for a right labium tumefaction measuring 15 cm. After excision, histologic examination revealed a well-circumscribed and lobulated tumor. The lobules were composed of slender spindle cells showing slightly eosinophilic cytoplasm with indistinct boundaries, uniform nuclei with finely granular chromatin and no nucleoli. Scattered Signet-ring-type lipoblasts were observed. The background was myxoid, with a "chickenwire" capillary vascular network mimicking a myxoid liposarcoma. No mitotic figures, necrosis or pleomorphism was noted. CONCLUSION: This mesenchymal tumor had adipocytic differentiation and no patent sign of malignancy, akin to infantile lipoblastoma. Because of the benign behavior of this tumor, complete excision without radiation is recommended. PMID- 17465292 TI - Mirror syndrome associated with sacrococcygeal teratoma: a case report. AB - BACKGROUND: Mirror syndrome is associated with both nonimmune and immune hydrops fetalis. The clinical manifestations are quite varied, and the pathophysiology is poorly understood. We describe a case of mirror syndrome associated with afetus that had a rapidly growing sacrococcygeal teratoma (SCT) without overt hydrops. CASE: At 30 weeks' gestational age a fetus with SCT began to show early sonographic evidence of right heart failure, placentomegaly and polyhydramnios without overt fetal hydrops. Shortly after these findings were noted, the mother began to develop hypertension, epigastric pain, proteinuria and thrombocytopenia. These findings were all reversed after delivery of the fetus. Subsequent surgery on the infant was successful. CONCLUSION: Mirror syndrome has been linked with SCT and is usually associated with severe fetal hydropic changes. In our case the development of mirror syndrome preceded the manifestations of overt hydrops. Identification of early signs of fetal compromise or hydrops may help to predict patients who will develop mirror syndrome and improve outcomes with earlier intervention. PMID- 17465293 TI - Maternal and neonatal death from advanced choriocarcinoma due to a delay in diagnosis: a case report. AB - BACKGROUND: Metastatic choriocarcinoma rarely coexists with a normal pregnancy. CASE: A 36-year-old multipara presented to the hospital at 23 weeks' gestation with progressive dyspnea and tachycardia. She was admitted and evaluated by the Psychiatry Department for presumed panic disorder and had perinatology, pulmonary, cardiology and infectious disease consultations. Over the course of 5 days she developed respiratory failure, delivered a nonviable infant and died from complications of metastatic pulmonary choriocarcinoma. Choriocarcinoma was not considered in the differential diagnosis of the respiratory failure until the day prior to the patient's death. CONCLUSION: Choriocarcinoma, although rare, should be considered in the differential diagnosis of pregnant women presenting with abnormally elevated serum beta-human chorionic gonadotropin levels and respiratory failure. PMID- 17465294 TI - Cesarean birth in the morbidly obese woman: a report of 3 cases. AB - BACKGROUND: Primary and repeat cesarean births are a frequent occurrence among morbidly obese women. Technical difficulties encountered in caring for these patients, coupled with physiologic differences, affect their operative management. CASES: Three morbidly obese women (190-296 kg, body mass index 56.7 93.6) had cesarean births utilizing a supraumbilical incision and internal retention abdominal wall closures. Alternative anesthetic management was required in 2 patients. Bariatric operative and postoperative equipment was required in each case, and varying thromboprophylaxis strategies were employed. CONCLUSION: Careful planning along with appropriate specialty consultation is required for a safe and successful cesarean birth in the morbidly obese woman. PMID- 17465295 TI - Rupture of a tuboovarian abscess into the anterior abdominal wall: a case report. AB - BACKGROUND: Rupture of a tuboovarian abscess is a life-threatening emergency. The current standard of care is surgery to manage the onset of peritonitis and sepsis. CASE: A 34-year-old woman presented with gradually worsening abdominal pain over several weeks. She had fever, pelvic tenderness, an elevated white blood cell count and ultrasonographic evidence of a tuboovarian abscess. After 48 hours of triple antibiotic therapy without a clinical improvement, she underwent surgery, during which she was found to have an abscess that had ruptured into the abdominal wall. A supracervical hysterectomy/bilateral salpingo-oophorectomy and surgical debridement of the subcutaneous tissue was performed, with wet to dry dressing changes of the surgical wound. Over the following 18 months, the patient required extensive wound care for recurrent bouts of wound drainage. CONCLUSION: Rupture of a tuboovarian abscess usually presents with sudden worsening of the patient's condition. Delays in surgical debridement and drainage increase the rate of associated mortality. Atypical presentations may present as cases resistant to conventional medical therapy, with fewer of the associated risks of life-threatening peritonitis and subsequent sepsis. Our patient represents the first reported case of extraperitoneal spillage of abscess contents from rupture of a tuboovarian abscess into the anterior abdominal wall. PMID- 17465296 TI - Double vagina and cervix communicating bilaterally with a single uterine cavity: report of a case with an unusual congenital uterine malformation. AB - BACKGROUND: The existence of a longitudinal vaginal septum with double cervix communicating bilaterally with a nonseptate uterine body and normal adnexa is an unusual mullerian anomaly. CASE: A 43-year-old woman presented with menorrhagia and duplication of the cervix and vagina. Afibromatous uterus was suggested by clinical examination and confirmed by ultrasonography. The patient underwent total abdominal hysterectomy with bilateral salpingooophorectomy. The surgical specimen revealed a fibromatous uterus with double cervix communicating bilaterally with a nonseptate uterine body; both adnexa were normal. CONCLUSION: This rare mullerian anomaly is inconsistent with the classical embryologic theory of caudal to cranial mullerian development but supports the alternative embryologic hypothesis suggested by Muller et al, according to which fusion and absorption begin at the isthmus and proceed simultaneously in both the cranial and caudal directions. PMID- 17465297 TI - Simultaneous ovarian and endometrial osseous metaplasia: a case report. AB - BACKGROUND: Endometrial ossification is a rare disease. More than 80% of cases occur after pregnancy, but it has been observed in patients with a history of endometritis, dilation and curettage, and metabolic disorders. CASE: A 42-year old woman presented with osseous metaplasia of both the endometrium and ovaries. At laparoscopy both adnexa were covered with adhesions and were adherent to the posterior wall of the uterus. Following adhesiolysis, calcified nodules were removed from both ovaries with biopsy forceps. Endometrial bone tissue was removed by hysteroscopic resection. CONCLUSION: To our knowledge, this is the first reported case of osseous metaplasia of both the endometrium and ovaries since all cases described to date in the literature involved only the uterine cavity. Conservative management with endoscopic surgery is effective. PMID- 17465298 TI - Antenatal diagnosis of meconium peritonitis in dichorionic twins: a case report. AB - BACKGROUND: Meconium peritonitis (MP) is a chemical peritonitis, and little is known about its natural history. Below we report MP in both fetuses from a twin pregnancy, with identical clinical ultrasonographic findings but a delay in presentation. CASE: A 31-year-old woman with a twin pregnancy was referred to our institution due to fetal bowel dilatation and ascites. Serial ultrasonography demonstrated gradual but dynamic changes. Both infants were delivered at 29 weeks'gestation. The larger infant had type II MP and experienced more postoperative complications and longer ventilatory care, parenteral nutrition and hospital stay than the smaller one, who had type III MP. CONCLUSION: The type of MP at birth may be more important than birth weight in terms of neonatal morbidity. This case increases our understanding of the natural history and prognosis of MP. PMID- 17465299 TI - Huge teratoma of the fallopian tube: a case report. AB - BACKGROUND: Primary mature solid teratomas of the fallopian tube are extremely rare and only 11 cases have been reported in the literature. A woman presented with a mature, solid teratoma of the fallopian tube, possibly the largest solid teratoma on record. CASE: A 24-year-old, nulliparous woman presented with abdominal distention, intermittent abdominal pain, nausea and vomiting for 4 weeks and was admitted to the hospital. Abdominal examination revealed a palpable, nontender mass. Computed tomography showed a hypodense lesion in the abdomen. Serum concentrations of carcinoembryonic antigen (2.6 ng/mL) and beta human chorionic gonadotropin (0.1 mIU/mL) were within normal limits, while serum levels of CA-125 (130 U/mL) and CA-19.9 (237 U/mL) were elevated. A multilobulated, pedunculated, mature, solid teratoma of the fallopian tube measured 31 x 21 x 14 cm and weighed 2,400 g. At laparotomy, it was located in the fimbrial portion of the left fallopian tube, and a left salpingectomy was performed. The tumor consisted of bone, hair, and sebaceous and sweat glands. The cavity of the cyst was lined mainly with skin composed of keratinized squamous epithelium and contained abundant sebaceous and sweat glands. CONCLUSION: In cases of undetermined pelvic or abdominal masses, especially with calcification, a teratoma of the fallopian tube should be considered. PMID- 17465300 TI - Cervical heterotopic pregnancy after assisted reproductive technology successfully treated with only simple embryo aspiration: a case report. AB - BACKGROUND: The etiology of cervical heterotopic pregnancy is unknown, but most are associated with assisted reproductive techniques. Various types of conservative management to save the intrauterine pregnancy have been attempted. CASE: A 35-year-old woman conceived after in vitro fertilization/embryo transfer for primary malefactor infertility. At 7(3/7) weeks of gestation, only the embryo was aspirated without fluid. Delivery of a healthy infant at 35 weeks was successful. CONCLUSION: Simple embryo aspiration under transvaginal ultrasonography guidance can be used in cervical heterotopic pregnancy. PMID- 17465301 TI - Spontaneous resolution of severe fetal ascites in the second trimester: a case report. AB - BACKGROUND: Fetal ascites is an infrequent anomaly. The prognosis for fetal and neonatal survival is poor, especially with the development of ascites before 24 weeks' gestation. The spontaneous remission of severe fetal ascites without intrauterine treatment, especially with an uncomplicated neonatal outcome, is extremely rare. CASE: A woman had sonographically documented fetal ascites at 19 weeks' gestation. The workup for immunologic and nonimmunologic causes was negative. Serial ultrasound examination showed ascites from 19 to 28 weeks' gestation. A complete resolution of the ascites was noted at 33 weeks' gestation. A normal male infant was vaginally delivered at 39 weeks' gestation. CONCLUSION: Spontaneous resolution of fetal ascites, with a good prognosis, can occur in cases with an idiopathic etiology. PMID- 17465302 TI - White spot syndrome virus envelope protein VP53A interacts with Penaeus monodon chitin-binding protein (PmCBP). AB - White spot syndrome virus (WSSV) is the causative agent of a severe disease of cultivated shrimp. Using purified WSSV virions, VP53A encoded by open reading frame wssv067 was identified as a structural protein by SDS-PAGE and proteomics. Immunoelectron microscopy with a gold-labeled secondary antibody revealed that VP53A was distributed on the viral envelope. In order to further explore the link between WSSV067 and host proteins, we performed a yeast 2-hybrid screening of a Penaeus monodon cDNA library, using WSSV067C as bait. One of the molecules that specifically interacted with WSSV067C was the P. monodon chitin-binding protein (PmCBP). An in vitro binding assay showed that c-myc-WSSV067C was capable of co precipitating HA-PmCBP-C. Furthermore, PmCBP was expressed in almost all organs but appeared to be up-regulated at the late stage of WSSV infection. PMID- 17465303 TI - Identification of icp11, the most highly expressed gene of shrimp white spot syndrome virus (WSSV). AB - This study investigates white spot syndrome virus (WSSV) gene expression levels in the cells of 2 hosts (Penaeus monodon and Litopenaeus vannamei). Microarray and expressed sequence tag (EST) analysis of the mRNA profiles in WSSV-infected P. monodon cells were used to identify WSSV genes that were very highly expressed. Results showed that the mRNA of the WSSV icp11 gene consistently had the highest copy number of all (3x higher than the major envelope protein, VP28). At the protein level in WSSV-infected L. vannamei, 2-dimensional gel analysis and liquid chromatography-nano-electrospray ionization tandem mass spectrometry (LC nanoESI-MS/MS) protein identification also showed that this WSSV non-structural protein has the highest expression levels reported to date. ICP11 is capable of self-multimerization, and it becomes located in both the cytoplasm and nucleus of the host cell. These data suggest that ICP11 plays an important, but presently unknown, role during viral infection, and that expression of the WSSV icp11 gene/WSSV ICP11 protein is potentially a good and diagnostically useful indicator of WSSV infection. PMID- 17465304 TI - Serological, virological and histopathological study of an outbreak of sleeping disease in farmed rainbow trout Oncorhynchus mykiss. AB - A prospective longitudinal survey for sleeping disease (SD) was carried out over a 20 wk period on a caged freshwater population of farmed rainbow trout Oncorhynchus mkyiss. Pancreas, heart and red and white skeletal muscle were examined histologically and the presence and severity of lesions recorded. Sera were tested for viraemia with Salmonid Alphavirus (SAV) and for virus neutralizing (VN) antibodies. Viraemia was detected for 4 wk, beginning at Week 6 and with a peak prevalence of 57.9% at Week 7. Clinical signs and mortalities appeared at Week 8. Total mortality in the study cage from Week 6 onward was 6.3 %, but other cages at the site had mortality levels of up to 47.2%. VN antibodies were first detected at Week 9, with seroprevalence increasing to 80% by the end of the study (Week 20). Geometric mean antibody titres peaked at 1/89.4 at Week 17. Histological lesions were first detected at Week 7 (pancreas only), before increasing in prevalence and severity to peak at Weeks 9 and 10. The majority of lesions were resolved by Week 15. PMID- 17465305 TI - Multiplex-PCR for simultaneous detection of 3 bacterial fish pathogens, Flavobacterium columnare, Edwardsiella ictaluri, and Aeromonas hydrophila. AB - A multiplex PCR (m-PCR) method was developed for simultaneous detection of 3 important fish pathogens in warm water aquaculture. The m-PCR to amplify target DNA fragments from Flavobacterium columnare (504 bp), Edwardsiella ictaluri (407 bp) and Aeromonas hydrophila (209 bp) was optimized by adjustment of reaction buffers and a touchdown protocol. The lower detection limit for each of the 3 bacteria was 20 pg of nucleic acid template from each bacteria per m-PCR reaction mixture. The sensitivity threshold for detection of the 3 bacteria in tissues ranged between 3.4 x 10(2) and 2.5 x 10(5) cells g(-1) of tissue (channel catfish Ictalurus punctatus Rafinesque). The diagnostic sensitivity and specificity of the m-PCR was evaluated with 10 representative isolates of each of the 3 bacteria and 11 other Gram-negative and 2 Gram-positive bacteria that are taxonomically related or ubiquitous in the aquatic environment. Except for a single species (A. salmonicida subsp. salmonicida), each set of primers specifically amplified the target DNA of the cognate species of bacteria. m-PCR was compared with bacteriological culture for identification of bacteria in experimentally infected fish. The m-PCR appears promising for the rapid, sensitive and simultaneous detection of Flavobacterium columnare, E. ictaluri and A. hydrophila in infected fish compared to the time-consuming traditional bacteriological culture techniques. PMID- 17465306 TI - Real-time PCR for quantification of viable Renibacterium salmoninarum in chum salmon Oncorhynchus keta. AB - Quantification of msa gene mRNA of Renibacterium salmoninarum, the causative agent of bacterial kidney disease (BKD), was investigated using reverse transcription followed by real-time PCR assay on R. salmoninarum in culture, and in experimentally challenged chum salmon Oncorhynchus keta fry kidney tissues (total of 70 samples) after intraperitoneal (i.p.) injection and bath infection. Correlations of msa gene mRNA concentrations with culturable cell concentrations (as colony forming units [CFU]), determined by drop-plate culture method on selective kidney disease medium (SKDM) agar through a 12 wk incubation time, and msa gene DNA concentrations by real-time PCR assay were examined. Furthermore, ovarian fluid samples from wild chum salmon adults with no clinical signs of disease were collected from 8 rivers and from clinically infected kokanee 0. nerka and masu salmon O. masou that were reared in 1 and 2 hatcheries, respectively (total of 414 samples). All samples were examined by nested PCR assay. Then, positive samples were examined by real-time PCR assays for mRNA and DNA; mRNA was detectable at 8 log units (5.0 x 101 to 5.0 x 10(9) copies p11(-1)) with high correlation (R2 = 0.999). The mRNA concentration correlated with CFU in kidney tissue from fish infected by i.p. injection (R2 = 0.924), by bath infection (R2 = 0.502) and in culture (R2 = 0.888). R. salmoninarum was detected and quantified by real-time PCR assay for mRNA in ovarian fluid samples in both subclinically infected chum salmon adults and clinically infected kokanee and masu salmon adults; detection rates ranged from 0 to 44.4% and concentrations ranged from 9.7 x 10(2) to 5.6 x 10(5) copies pl(-1). These results indicate that real-time PCR assay for the mRNA is a rapid, sensitive and reliable method to detect and quantify the viability of R. salmoninarum in kidney and ovarian fluid samples of salmonid fishes with both clinical and subclinical infection of the pathogen. PMID- 17465307 TI - Parasite fauna of bream Abramis brama and roach Rutilus rutilus from a man-made waterway and a freshwater habitat in northern Germany. AB - Fifty specimens each of bream Abramis brama and roach Rutilus rutilus were examined for metazoan parasite fauna and trichodinid ciliates; 25 specimens of each species were collected from the Kiel Canal, a man-made waterway, and a nearby freshwater lake, the Dieksee. This is the first detailed parasitological examination of A. brama and R. rutilus at these locations: 30 parasite species were found, comprising 4 protozoans, 4 myxozoans, 5 digeneans, 3 monogeneans, 2 cestodes, 6 nematodes, 2 acanthocephalans, 3 crustaceans and 1 hirudinean. The crustacean Caligus lacustris occurred in both habitats while 2 other crustacean species, 2 acanthocephalans and 1 hirudinean were recorded exclusively for the lake habitat. Larval as well as adult stages of the different parasite species were found, indicating that both fish species act as intermediate and final hosts in both habitats. The Kiel Canal (total of 17 parasite species) showed a lower parasite species richness for A. brama and R. rutilus (14 and 10 parasite species, respectively) than the lake (25 parasite species). A. brama had a higher parasite richness (22 species) than R. rutilus (16 species) in the lake habitat. Most parasites collected were of freshwater origin. Consequently, the observed infection pattern of both fish species in the waterway is mainly influenced by the limited salinity tolerance of freshwater parasites, which are negatively affected even by a salinity of 2.3 to 4.5. In the central Kiel Canal, neither fish species was infected with marine parasites of low host specifity. These parasites are either limited by the low salinity at this sampling site (<4.5 to 6.0) or they cannot enter the canal due to the environmental conditions prevailing in this artificial brackish water habitat. Thus, the canal may comprise a natural barrier preventing the distribution of North Sea parasites into the Baltic Sea. However, the brackish water Baltic Sea nematodes Paracuaria adunca and Cosmocephalus obvelatus were found in R. rutilus from the canal, demonstrating the ability of some parasite species to invade and extend their range of distribution through this man-made shipping route from the Baltic to the North Sea. PMID- 17465308 TI - Effect of water temperature on reproductive development of Benedenia seriolae (Monogenea: Capsalidae) from Seriola lalandi in Australia. AB - The monogenean Benedenia seriolae (Yamaguti, 1934) Meserve, 1938 is a major pathogen of farmed yellowtail kingfish Seriola lalandi in South Australia. To control parasite populations in commercial farms, an understanding of the effect of water temperature on parasite development is vital. This study investigated the effect of water temperature (14, 18, 22 and 26 +/- 0.5 degrees C) on development, growth rates and age at sexual maturity of B. seriolae. Five distinct developmental stages, defined by the progressive development of reproductive organs, were used to describe development of B. seriolae from recently invaded larvae to sexually mature adults. Parasite age at the first sign of sexual maturity (ability to lay eggs) was strongly influenced by water temperature and was attained at 41, 24, 16 and 14 d post infection (p.i.) at 14, 18, 22 and 26 +/- 0.5 degrees C respectively. Four parameters (parasite total length, maximum parasite width, accessory sclerite length and anterior hamulus length) were examined for suitability as an index of parasite age. Growth rates for each parameter increased with water temperature; however, mean anterior hamulus length was identified as the most reliable index of parasite age. Equations derived from these data can be used to estimate parasite age and time to sexual maturity at water temperatures ranging from 14 to 26 degrees C. The equations provide a simple tool to assist implementation of strategic treatment plans for B. seriolae infections in commercial kingfish farms, not only in South Australia but in other localities worldwide where S. lalandi is farmed. PMID- 17465309 TI - Synechocystis MCCB 114 and 115 as putative probionts for Penaeus monodon post larvae. AB - Synechocystis MCCB 114 and 115 were segregated as putative probionts for shrimp larvae from a collection of 54 cyanobacterial cultures enriched from seawater. On feeding Penaeus monodon post-larvae with the cyanobacteria, the generic diversity of the intestinal bacterial flora could be enhanced with substantial reduction or total absence of Vibrio spp. A significant difference (p < 0.001) in the percent survival of batches of post-larvae fed on the cyanobacterial cultures was observed and, on repeated challenge with V. harveyi, the relative percent survival of those batches of larvae fed on Synechocystis MCCB 114 and 115 was significantly higher. The Synechocystis MCCB 114 and 115 cultures were found to contain high levels of protein (34 to 43%), in addition to carotenoids. PMID- 17465310 TI - Scuticociliate infection and pathology in cultured turbot Scophthalmus maximus from the north of Portugal. AB - During the years 2004 and 2005 high mortalities in turbot Scophthalmus maximus (L.) from a fish farm in the north of Portugal were observed. Moribund fish showed darkening of the ventral skin, reddening of the fin bases and distended abdominal cavities caused by the accumulation of ascitic fluid. Ciliates were detected in fresh mounts from skin, gill and ascitic fluid. Histological examination revealed hyperplasia and necrosis of the gills, epidermis, dermis and muscular tissue. An inflammatory response was never observed. The ciliates were not identified to species level, but the morphological characteristics revealed by light and electronic scanning microscopes indicated that these ciliates belonged to the order Philasterida. To our knowledge this is the first report of the occurrence of epizootic disease outbreaks caused by scuticociliates in marine fish farms in Portugal. PMID- 17465311 TI - Is matching innate? AB - Experimentally naive mice matched the proportions of their temporal investments (visit durations) in two feeding hoppers to the proportions of the food income (pellets per unit session time) derived from them in three experiments that varied the coupling between the behavioral investment and food income, from no coupling to strict coupling. Matching was observed from the outset; it did not improve with training. When the numbers of pellets received were proportional to time invested, investment was unstable, swinging abruptly from sustained, almost complete investment in one hopper, to sustained, almost complete investment in the other-in the absence of appropriate local fluctuations in returns (pellets obtained per time invested). The abruptness of the swings strongly constrains possible models. We suggest that matching reflects an innate (unconditioned) program that matches the ratio of expected visit durations to the ratio between the current estimates of expected incomes. A model that processes the income stream looking for changes in the income and generates discontinuous income estimates when a change is detected is shown to account for salient features of the data. PMID- 17465312 TI - The effects of reinforcer magnitude on timing in rats. AB - The relation between reinforcer magnitude and timing behavior was studied using a peak procedure. Four rats received multiple consecutive sessions with both low and high levels of brain stimulation reward (BSR). Rats paused longer and had later start times during sessions when their responses were reinforced with low magnitude BSR. When estimated by a symmetric Gaussian function, peak times also were earlier; when estimated by a better-fitting asymmetric Gaussian function or by analyzing individual trials, however, these peak-time changes were determined to reflect a mixture of large effects of BSR on start times and no effect on stop times. These results pose a significant dilemma for three major theories of timing (SET, MTS, and BeT), which all predict no effects for chronic manipulations of reinforcer magnitude. We conclude that increased reinforcer magnitude influences timing in two ways: through larger immediate after-effects that delay responding and through anticipatory effects that elicit earlier responding. PMID- 17465313 TI - Tests of behavioral-economic assessments of relative reinforcer efficacy: economic substitutes. AB - This experiment was conducted to test predictions of two behavioral-economic approaches to quantifying relative reinforcer efficacy. According to the first of these approaches, characteristics of averaged normalized demand curves may be used to predict progressive-ratio breakpoints and peak responding. The second approach, the demand analysis, rejects the concept of reinforcer efficacy, arguing instead that traditional measures of relative reinforcer efficacy (breakpoint, peak response rate, and choice) correspond to specific characteristics of non-normalized demand curves. The accuracy of these predictions was evaluated in rats' responding for food or fat: two reinforcers known to function as partial substitutes. Consistent with the first approach, predicted peak normalized response output values (Omax) obtained under single schedule conditions ordinally predicted progressive-ratio breakpoints and peak responding. Predictions of the demand analysis had mixed success. Pmax and Omax were significantly correlated with PR breakpoints and peak responding (respectively) when fat, but not when food, was the reinforcer. Relative consumption of food and fat under single schedules of reinforcement did not predict preference better than chance. The normalized demand analysis is supplemented with the economic concept of diminishing marginal utility, to predict preference shifts across the range of food and fat prices examined. PMID- 17465314 TI - Local effects of delayed food. AB - Five pigeons were trained on a procedure in which seven concurrent variable interval schedules arranged seven different food-rate ratios in random sequence in each session. Each of these components lasted for 10 response-produced food deliveries, and components were separated by 10-s blackouts. We varied delays to food (signaled by blackout) between the two response alternatives in an experiment with three phases: In Phase 1, the delay on one alternative was 0 s, and the other was varied between 0 and 8 s; in Phase 2, both delays were equal and were varied from 0 to 4 s; in Phase 3, the two delays summed to 8 s, and each was varied from 1 to 7 s. The results showed that increasing delay affected local choice, measured by a pulse in preference, in the same way as decreasing magnitude, but we found also that increasing the delay at the other alternative increased local preference. This result casts doubt on the traditional view that a reinforcer strengthens a response depending only on the reinforcer's value discounted by any response-reinforcer delay. The results suggest that food guides, rather than strengthens, behavior. PMID- 17465315 TI - Stimulus control and compounding with ambient odor as a discriminative stimulus on a free-operant baseline. AB - Previous experiments have demonstrated that the simultaneous presentation of independently established discriminative stimuli can control rates of operant responding substantially higher than the rates occasioned by the individual stimuli. This "additive summation" phenomenon has been shown with a variety of different reinforcers (e.g., food, water, shock avoidance, cocaine, and heroin). Discriminative stimuli previously used in such studies have been limited to the visual and auditory sensory modalities. The present experiment sought to (1) establish stimulus control on a free-operant baseline with an ambient olfactory discriminative stimulus, (2) compare olfactory control to that produced with an auditory discriminative stimulus, and (3) determine whether compounding independently established olfactory and auditory discriminative stimuli produces additive summation. Rats lever pressed for food on a variable-interval schedule in the presence of either a tone or an odor, with comparable control developed to each stimulus. In the absence of these stimuli responding was not reinforced. During stimulus compounding tests, the tone-plus-odor compound occasioned more than double the responses occasioned by either the tone or odor presented individually. Thus, the current study (1) established stimulus control with an ambient olfactory discriminative stimulus in a traditional free-operant setting and (2) extended the generality of stimulus-compounding effects by demonstrating additive summation when olfactory and auditory discriminative stimuli were presented simultaneously. PMID- 17465316 TI - Preference for a stimulus that follows a relatively aversive event: contrast or delay reduction? AB - Several types of contrast effects have been identified including incentive contrast, anticipatory contrast, and behavioral contrast. Clement, Feltus, Kaiser, and Zentall (2000) proposed a type of contrast that appears to be different from these others and called it within-trial contrast. In this form of contrast the relative value of a reinforcer depends on the events that occur immediately prior to the reinforcer. Reinforcers that follow relatively aversive events are preferred over those that follow less aversive events. In many cases the delay reduction hypothesis proposed by Fantino (1969) also can account for such effects. The current experiments provide a direct test of the delay reduction and contrast hypotheses by manipulating the schedule of reinforcement while holding trial duration constant. In Experiment 1, preference for fixed interval (FI) versus differential-reinforcement-of-other-behavior (DRO) schedules of reinforcement was assessed. Some pigeons preferred one schedule over the other while others demonstrated a position (side) preference. Thus, no systematic preference was found. In Experiment 2, a simultaneous color discrimination followed the FI or DRO schedule, and following training, preference was assessed by presenting the two positive stimuli simultaneously. Consistent with the contrast hypothesis, pigeons showed a significant preference for the positive stimulus that in training had followed their less preferred schedule. PMID- 17465318 TI - The analysis of behavior: what's in it for us? AB - When we publish behavioral research, we are not allowed to communicate the thrill, the poetry, or the exhilaration that are outcomes of the discovery process. Yet, these are among our most potent reinforcers. Explicit recognition of the emotional accompaniments to research could help attract students into the experimental analysis of behavior. PMID- 17465317 TI - New knowledge derived from learned knowledge: functional-anatomic correlates of stimulus equivalence. AB - Forming new knowledge based on knowledge established through prior learning is a central feature of higher cognition that is captured in research on stimulus equivalence (SE). Numerous SE investigations show that reinforcing behavior under control of distinct sets of arbitrary conditional relations gives rise to stimulus control by new, derived relations. This investigation examined whether frontal-subcortical and frontal-parietal networks known to support reinforced conditional relations also support derived conditional relations. Twelve adult subjects completed matching-to-sample (MTS) training with correct/wrong feedback to establish four trained conditional relations within two distinct, three-member stimulus classes: (1) A1-->B1, B1-->C1 and (2) A2-->B2, B2-->C2. Afterwards, functional neuroimaging was performed when MTS trials were presented involving matching two identical circles (a sensorimotor control condition), trained relations (A-->B, B-->C), and derived relations: symmetry (B-->A, C-->B), transitivity (A-->C), and equivalence (C-->A). Conditional responding to trained and derived relations was similarly correlated with bilateral activation in the targeted networks. Comparing trained to derived relations, however, highlighted greater activation in several prefrontal regions, the caudate, thalamus, and putamen, which may represent the effects of extended training or feedback present during imaging. Each derived relation also evidenced a unique activation pattern. Collectively, the findings extend the role of frontal-subcortical and frontal parietal networks to derived conditional relations and suggest that regional involvement varies with the type of derived conditional relation. PMID- 17465319 TI - The symmetry of man. AB - The paper contains a description of basic regularities in the manifestation of symmetry of human structural organization and its ontogenetic and phylogenetic development. A concept of macrobiocrystalloid with inherent complex symmetry is proposed for the description of the human organism in its integrity. The symmetry can be characterized as two-plane radial (quadrilateral), where the planar symmetry is predominant while the layout of organs of radial symmetry is subordinated to it. Out of the two planes of symmetry (sagittal and horizontal), the sagittal plane is predominant. The symmetry of the chromosome, of the embrio at the early stages of cell cleavage as well as of some organs and systems in their phylogenetic development is described. An hypothesis is postulated that the two-plane symmetry is formed by two mechanisms: a) the impact of morphogenetic fields of the whole crystalloid organism during embriogenesis and, b) genetic mechanisms of the development of chromosomes having two-plane symmetry. PMID- 17465320 TI - The biocrystalloid structure of man: an extracellular theory. AB - Medical science based solely on the postulates of the cellular theory does not describe the structure and functions of biological organisms in full or meet the needs of modern medicine. Biological organisms, man included, can be regarded as biocrystalloids that are composites of crystals and paracrystalline media at the level of the whole organism and individual cells alike. The core component of biocrystalloids is represented by extracellular components of the connective tissue. The paper describes major features of biocrystalloids and highlights their similarity to mineral organisms in which a growing or weathering crystal also regulates the flows of the paracrystalline medium. The paper concludes that further research into the force fields of a biocrystalloid will allow developing new diagnostic and treatment modalities. PMID- 17465321 TI - Reconstruction in space and visualization of a planar image: a mathematical and computational introduction. AB - Aim of this paper is to provide a short introduction to the fundamental mathematical ideas involved in tri-dimensional (3D) image reconstruction, in a basic way as to make such ideas available to a large audience. Particular attention is then given to the case in which the bi-dimensional (2D) input data are represented of very few images. A feasibility study is outlined for a 3D reconstruction of a human gland or organ, satisfying hypothesis of uniformity in the tissue and in the arterial distribution, spacial extension and density. The resulting method aims at obtaining functional information on the target organ, minimizing the number (and social cost) of possibly invasive imaging techniques by employing one or very few scintigraphic or echographic images. PMID- 17465322 TI - Complexity seems to open a way towards a new Aristotelian-Thomistic ontology. AB - Today's sciences seem to converge all towards very similar foundational questions. Such claims, both of epistemological and ontological nature, seem to rediscover, in a new fashion some of the most relevant topics of ancient Greek and Mediaeval philosophy of nature, logic and metaphysics, such as the problem of the relationship between the whole and its parts (non redictionism), the problems of the paradoxes arising from the attempt to conceive the entity like an univocal concept (analogy and analogia entis), the problem of the mind-body relationship and that of an adequate cognitive theory (abstraction and immaterial nature of the mind), the complexity of some physical, chemical and biological systems and global properties arising from information (matter-form theory), etc. Medicine too is involved in some of such relevant questions and cannot avoid to take them into a special account. PMID- 17465323 TI - Finite machines, mental procedures, and modern physics. AB - A Turing machine provides a mathematical definition of the natural process of calculating. It rests on trust that a procedure of reason can be reproduced mechanically. Turing's analysis of the concept of mechanical procedure in terms of a finite machine convinced Godel of the validity of the Church thesis. And yet, Godel's later concern was that, insofar as Turing's work shows that "mental procedure cannot go beyond mechanical procedures", it would imply the same kind of limitation on human mind. He therefore deems Turing's argument to be inconclusive. The question then arises as to which extent a computing machine operating by finite means could provide an adequate model of human intelligence. It is argued that a rigorous answer to this question can be given by developing Turing's considerations on the nature of mental processes. For Turing such processes are the consequence of physical processes and he seems to be led to the conclusion that quantum mechanics could help to find a more comprehensive explanation of them. PMID- 17465324 TI - Perception in action. AB - For long time perception and action have been considered as separated and peripheral processes, devoid of any intrinsic cognitive value. Over the last years, however, this view has been challenged by several neurophysiological data. The aim of this paper is to provide an overview of some recent studies on the cortical visual and motor systems. In particular, we'll focus on the discovery (in the inferior parietal lobe and in the premotor cortex) of two different kinds of sensorimotor neurons that not only are involved in the motor control of actions, but also play an essential role in object categorizing and in action understanding. We'll argue that the functional properties of these kinds of neurons show the limits and abstractness of any strict dichotomy between perception and action, and pave the way to a requalification of some relevant philosophical perspectives on these issues. PMID- 17465325 TI - Synaptic plasticity and the neurobiology of learning and memory. AB - Learning and memory are fundamental higher brain functions that allow the individual to adapt to the environment, to build up his own history as a unique creature, to widen the personal cultural background and, ultimately, the population culture. In this review, we will briefly examine the cellular and molecular mechanisms that contribute to the various forms of memory that include short- and long-term memory as well as unconscious and conscious memory. Although in mammals various brain areas participate in distinct forms of memory, the molecular and cellular mechanisms of very simple to complex forms of learning and memory are extremely conserved across evolution from molluscs to man and among various forms of memory and consist in short-to-long lived rearrangements in synaptic efficiency and in the structure of neuronal networks. PMID- 17465326 TI - Computation and brain processes, with special reference to neuroendocrine systems. AB - The development of neural networks and brain automata has made neuroscientists aware that the performance limits of these brain-like devices lies, at least in part, in their computational power. The computational basis of a. standard cybernetic design, in fact, refers to that of a discrete and finite state machine or Turing Machine (TM). In contrast, it has been suggested that a number of human cerebral activites, from feedback controls up to mental processes, rely on a mixing of both finitary, digital-like and infinitary, continuous-like procedures. Therefore, the central nervous system (CNS) of man would exploit a form of computation going beyond that of a TM. This "non conventional" computation has been called hybrid computation. Some basic structures for hybrid brain computation are believed to be the brain computational maps, in which both Turing like (digital) computation and continuous (analog) forms of calculus might occur. The cerebral cortex and brain stem appears primary candidate for this processing. However, also neuroendocrine structures like the hypothalamus are believed to exhibit hybrid computional processes, and might give rise to computational maps. Current theories on neural activity, including wiring and volume transmission, neuronal group selection and dynamic evolving models of brain automata, bring fuel to the existence of natural hybrid computation, stressing a cooperation between discrete and continuous forms of communication in the CNS. In addition, the recent advent of neuromorphic chips, like those to restore activity in damaged retina and visual cortex, suggests that assumption of a discrete continuum polarity in designing biocompatible neural circuitries is crucial for their ensuing performance. In these bionic structures, in fact, a correspondence exists between the original anatomical architecture and synthetic wiring of the chip, resulting in a correspondence between natural and cybernetic neural activity. Thus, chip "form" provides a continuum essential to chip "function". We conclude that it is reasonable to predict the existence of hybrid computational processes in the course of many human, brain integrating activities, urging development of cybernetic approaches based on this modelling for adequate reproduction of a variety of cerebral performances. PMID- 17465327 TI - Infundibular tanycytes as modulators of neuroendocrine function: hypothetical role in the regulation of the thyroid and gonadal axis. AB - Tanycytes comprise a heterogeneous population of specialized cells of glial origin that line the floor and ventrolateral walls of the third ventricle between the rostral and caudal limits of the hypothalamic median eminence. While morphologic and ultrastructural features suggest a role as barrier cells, creating separate compartments between the cerebrospinal fluid, median eminence and hypothalamus, tanycytes likely have multiple other important functions that have yet to be fully elucidated. Possibilities to consider are a role in neuroendocrine regulation including modulation of the hypothalamic-pituitary thyroid axis during fasting and infection, regulation of reproductive function, particularly in seasonal breeders, and in feeding. PMID- 17465328 TI - 13C NMR isotopomeric analysis and its application in the study of endocrine cell metabolism and function. AB - Defining mechanisms and enzymatic paths critical to cellular function (e.g., secretion) of endocrine cells is a key research goal that can lead toward novel avenues of therapeutic intervention for a variety of disorders. 13C NMR spectroscopy and isotopomer analysis of cell extracts are excellent tools to quantitatively assess metabolism through intermediate labeling and estimate carbon entry to the TCA cycle. Discussed are: cell lines and in vitro culturing; extraction of intracellular material; NMR spectroscopy of the extract; isotopomeric analysis and modeling to obtain relative metabolic fluxes to the TCA cycle. This paper describes issues related to the application of NMR spectroscopic techniques on cell line extracts. Included are results of two studies that illustrate considerations that must be taken when performing analogous studies on neuroendocrine tissue: one involving the effect of media composition on cell behavior and isotopomer labeling; the second looking at effects of applying different metabolic models to 13C data and inferences that may be drawn. NMR isotopomeric analysis is a powerful technique that may be applied to better understand endocrine cell function. PMID- 17465329 TI - Intranuclear 3'-phosphoinositide metabolism and apoptosis protection in PC12 cells. AB - Lipid second messengers, particularly those derived from the polyphosphoinositide metabolism, play a pivotal role in multiple cell signaling networks. Phosphoinositide 3-kinase (PI3K) generates specific 3'-phosphorylated inositol lipids that have been implicated in a multitude of cell functions. One of the best characterized targets of PI3K lipid products is the serine/threonine protein kinase Akt (protein kinase B). Recent findings have implicated the PI3K/Akt pathway in cancer progression because it stimulates cell proliferation and suppresses apoptosis. Evidence accumulated over the past 15 years has highlighted the presence of an autonomous nuclear inositol lipid cycle, and strongly suggests that lipid molecules are important components of signaling networks operating within the nucleus. PI3K, its lipid products, and Akt have also been identified at the nuclear level. In this review, we shall summarize the most updated findings about these molecules in relationship with suppression of apoptotic stimuli in PC12 cells. PMID- 17465330 TI - Serum peptide profiling: identifying novel cancer biomarkers for early disease detection. AB - Recent advances in mass spectrometry have enabled the identification of hundreds of low molecular weight (LMW) peptides that have previously been difficult to detect in human serum. Serum peptide patterns can now be analyzed using commercially available statistical programs to identify potential peptide patterns that may correlate with the presence or absence of specific diseases. A serum peptide profile (SPP), which is unique to each patient, can be created and compared to a known SPP from a specific disease. The SPP thus serves as a potential early stage biomarker prior to the clinical manifestation of disease. A unique and automated technology platform has been developed by members of the Protein Center at Memorial Sloan-Kettering Cancer Center (MSKCC). It involves a magnetic bead-based approach to extract LMW peptides from serum, placing them by robotic automation on a stainless steel MALDI-TOF target plate, subjecting them to mass spectrometric analysis, and using GeneSpring software to analyze the peptide patterns. Human serum from a cohort of 27 patients with metastatic thyroid cancer and 32 controls were analyzed on the MSKCC platform. 549 individual LMW peptides were identified. A SPP composed of 98 discriminatory LMW peptides was able to distinguish between the two groups of serum samples with high statistical accuracy. We believe that our automated system will serve as a model for future biotechnology laboratories in the quest for hidden diagnostic clues that may be detected by simply analyzing a drop of blood. PMID- 17465331 TI - The bioartificial thyroid: a biotechnological perspective in endocrine organ engineering for transplantation replacement. AB - A new concept for ex situ endocrine organ bioengineering is presented, focused on the realization of a human bioartificial thyroid gland. It is based on the theoretical assumption and experimental evidence that symmetries in geometrical coordinates of the thyroid tissue remain invariant with respect to developmental, physiological or pathophysiological transformations occuring in the gland architecture. This topological arrangement is dependent upon physical connections established between cells, cell adhesion molecules and extracellular matrix, leading to the view that the thyroid parenchyma behaves like a deformable "putty", moulded onto an elastic stromal/vascular scaffold (SVS) dictating the final morphology of the gland. In particular, we have raised the idea that the geometry of the SVS per se provides pivotal epigenetic information to address the genetically-programmed, thyrocyte and endothelial/vascular proliferation and differentiation towards a functionally mature gland, making organ form a pre requirementfor organ function. A number of experimental approaches are explored to obtain a reliable replica of a human thyroid SVS, and an informatic simulation is designed based on fractal growth of the thyroid intraparenchymal arterial tree. Various tissue-compatible and degradable synthetic or biomimetic polymers are discussed to act as a template of the thyroid SVS, onto which to co-seed autologous human thyrocyte (TPC) and endothelial/vascular (EVPC) progenitor cells. Harvest and expansion of both TPC and EVPC in primary culture are considered, with specific attention to the selection of normal thyrocytes growing at a satisfactory rate to colonize the synthetic matrix. In addition, both in vitro and in vivo techniques to authenticate TPC and EVPC lineage differentiation are reviewed, including immunocytochemistry, reverse trascriptase-polymerase chain reaction, flow cytomery and proteomics. Finally, analysis of viability of the thyroid construct following implantation in animal hosts is proposed, with the intent to obtain a bioartificial thyroid gland morphologically and functionally adequate for transplantation. We believe that the biotechnological scenario proposed herein may provide a template to construct other, more complex and clinically-relevant bioartificial endocrine organs ex situ, such as human pancreatic islets and the liver, and perhaps a new approach to brain bioengineering. PMID- 17465332 TI - Hypothesis: Musculin is a hormone secreted by skeletal muscle, the body's largest endocrine organ. Evidence for actions on the endocrine pancreas to restrain the beta-cell mass and to inhibit insulin secretion and on the hypothalamus to co ordinate the neuroendocrine and appetite responses to exercise. AB - Recent studies indicate that skeletal muscle may act as an endocrine organ by secreting interleukin-6 (IL-6) into the systemic circulation. From an analysis of the actions of IL-6 and of additional literature, we postulate that skeletal muscle also secretes an unidentified hormone, which we have named Musculin (Latin: musculus = muscle), which acts on the pancreatic beta-cell to restrain the size of the (beta-cell mass and to tonically inhibit insulin secretion and biosynthesis. It is suggested that the amount of Musculin secreted is determined by, and is positively correlated with, the prevailing insulin sensitivity of skeletal muscle, thereby accounting for the hyperinsulinemia that occurs in insulin resistant disorders such as type 2 diabetes mellitus, obesity, and the polycystic ovary syndrome. In addition, it is postulated that Musculin acts on the hypothalamus (arcuate nucleus, dorsomedial hypothalamic nucleus) to co ordinate the neuroendocrine and appetite responses to exercise. However, the possibilities that Musculin may act on additional central nervous system sites and that an additional hormone(s) may be responsible for these actions are not excluded. It is suggested that a search be made for Musculin, since analogues of such a substance may be of therapeutic benefit in the treatment of the current global diabetes and obesity epidemic. PMID- 17465333 TI - A pathogenic mechanism leading to partial lipodistrophy and prospects for pharmacological treatment of insulin resistance syndrome. AB - The understanding of a common complex phenotype such as insulin resistance can be favoured by evaluation of monogenic syndromes. Clinical definition, pathogenesis, and therapeutical strategies for the insulin resistance syndrome can thus be improved by the characterization at the molecular genetic level of monogenic forms of lipodystrophies. Here we report experimental evidence on the pathogenic mechanism underlying insulin resistance in a rare form of laminopathy, due to mutation of the LMNA gene coding for lamin A/C, the Dunnigan-type familial partial lipodystrophy (FPLD). The defect, consisting in the intranuclear accumulation of mutant unprocessed precursors of lamin A, reduces the amount of the DNA-bound adipocyte transcription factor sterol regulatory element binding protein 1 (SREBP1) and lowers the peroxisome proliferator-activated receptor (PPARgamma) expression, causing the impairment of pre-adipocyte differentiation. The treatment with the PPARgamma ligand troglitazone (TDZ) is able to rescue the adipogenic program. Since FPLD recapitulates the essential metabolic abnormalities of the common insulin resistance syndrome, the beneficial effects of TDZ on monogenic lipodystrophies might provide a clue as to the future treatment strategies also for the common syndrome of insulin resistance. PMID- 17465334 TI - Thyrotropin releasing hormone (TRH) may preserve pancreatic islet cell function: potential role in the treatment of diabetes mellitus. AB - Thyrotropin Releasing Hormone (TRH), first identified in the hypothalamus as a regulator of the Pituitary-Thyroid axis, has also been found in the beta-cell of the pancreas co-localised with insulin. The significance of this association is emphasised by the report that the TRH knock-out (KO) mouse is hyperglycemic. These findings have led to speculation that TRH may have a physiologic role in the regulation of carbohydrate metabolism. To understand better the role of TRH in the pancreas, TRH was administered to rats rendered diabetic from streptozotocin damage to the islets of Langerhans. This resulted in almost complete normalisation of the profound hyperglycemia. TRH is capable of reversing Diabetes Mellitus (DM) in an experimental animal model, possibly by promoting neogenesis of beta cells through induction of adult stem cells in the pancreas. These studies point to a potential therapeutic role for TRH in the treatment of DM in man. PMID- 17465335 TI - Frailty as a dysruption of steroid "syncrinology" in elderly man. AB - In the background of the endocrinological hypothesis of aging, frailty in man can be seen as a dysruption of syncrinology, a term with which we could define the harmonized activity of steroid hormones (DHEAS, cortisol, testosterone). Such endocrine age-related modifications might promote the activation of sarcopenia, which exerts a core position in the progression of frailty in the elderly. Consequently, the use of replacement treatment in order to delay the beginning of such steroid derangement might be a suitable strategy to improve the quality of life of man, whose life length has been significantly extended. PMID- 17465336 TI - Time for integration: communication in the immune system. AB - Signals exchanged between cells in the immune system are generated at specific contact regions termed "immunological synapses" (IS). The morphology and the functions of IS vary with circumstances and include directing secretion and integrating positive and negative signals to tune the extent of response - a model that is essentially derived from neurosciences. Indeed, the mechanisms of signal integration and antigen decoding are not yet completely understood and many hypotheses are worked out. PMID- 17465337 TI - Central nervous system recognition of peripheral inflammation: a neural, hormonal collaboration. AB - A key question in understanding communication between the immune system and the brain is: how does the central nervous system (CNS) recognise the extent of inflammatory or other immune responses taking place in peripheral tissues, so that it may respond appropriately? A variety of experimental models, particularly of fever, hypothalamus pituitary adrenal axis (HPAA) activation and sickness behaviour have been employed to investigate this. Although each of these responses is, at least in part, regulated in different areas of the CNS, they all require common information about peripheral inflammation in order to mount a coordinated response. Cytokines produced during the inflammatory response have been proposed as the important humoral mediators by which this is achieved. The experimental systems employed to investigate this have been dominated largely by examination of responses to bacterial lipopolysaccharide (LPS), which produces robust responses but does not make it easy to track down communication pathways when the LPS reaches the circulation. The data has been quite varied and several mediators and mechanisms have been considered responsible for interacting with the CNS, possibly because each plays a role at some level and their importance depends on the experimental system being examined. Both cytokines and LPS are able to induce the production of prostaglandins, which can in turn modify neural afferent pathways affecting CNS responses. Accompanying the variable dependence of these responses on neural regulation, the main endocrine input that arises from inflammatory tissues seems attributable to interleukin-6 (IL-6). This may gain direct access to the CNS but appears primarily to act by inducing the brain microvasculature to produce prostaglandin E2, which then alters the activity of neurons controlling body temperature, HPAA and behavioural responses. PMID- 17465338 TI - Pituitary autoantibodies in autoimmune polyendocrinopathy-candidiasis-ectodermal dystrophy (APECED). AB - Autoimmune polyendocrinopathy-candidiasis-ectodermal dystrophy (APECED) is an autosomal recessive disease due to mutations in the AIRE (AutoImmune REgulator) gene. The role of pituitary autoimmunity in APECED is not known. We determined the prevalence of pituitary autoantibodies in a cohort of 67 Finnish patients with APECED from 217 serum samples collected over 26 years by one investigator. Overall, autoantibodies to the 49 kDa cytosolic autoantigen, human pituitary enolase were detected in 39 of the 67 patients (58%). On their first sample, 25 patients had autoantibodies compared to 5 of 68 controls (chi-square, 1df=17.11, p< 0.001; OR=7.32), but subsequently 14 patients seroconverted between 10 and 53 years of age. Once seropositive, all but two of the patients maintained their positive autoantibody status, even over many years. In the current study all but 7 of the 19 patients known to have high titre anti-candidal enolase antibodies had developed autoantibodies directed against human pituitary enolase. Other pituitary autoantibody reactivities were detected against cytosolic proteins of molecular weights 40-, 45-, 60- and 105 kDa in 15%, 16%, 12% and 3% of patients respectively. Autoantibodies to pituitary enolase are markers of neuroendocrine autoimmunity but seem not to be associated with clinical hypopituitarism in APECED patients. PMID- 17465339 TI - Prolactin and autoimmune diseases in humans. AB - Prolactin has been shown to have immunomodulatory as well as lactogenic effects. Generally less well known is that prolactin may also play a role in the activity of autoimmune diseases such as systemic lupus erythematosus and rheumatoid arthritis. Studies have shown decreasing prolactin production to be beneficial in animal models of autoimmune disease. Thus far, double-blinded, placebo-controlled studies of dopamine agonist treatment in humans with autoimmune disease have been done only in lupus patients, and support the potential efficacy of such agents. Small, open-label trials have also suggested potential benefit in patients with rheumatoid arthritis, Reiter's syndrome, and psoriasis. More studies are required to further delineate the mechanisms by which prolactin affects autoimmune disease activity, to determine in which specific diseases prolactin plays a significant role, and to test the efficacy of prolactin-lowering agents as therapy for such diseases. PMID- 17465340 TI - A role of the TRAIL-TRAIL receptor system in the pathogenesis of diabetes. AB - The TNF-alpha super-family of cytokines comprises structurally related proteins that play pivotal roles in regulating cell death, immune response and inflammation. A new member of the family namely Tumor necrosis factor alpha Related Apoptosis-Inducing Ligand (TRAIL) is involved not only in apoptosis and immune regulation, but also it has a provocative role in vascular biology as reported recently. In this report we provide evidence that this new function of TRAIL may have a significance in the pathogenesis of diabetes and in particular in the vascular alterations that occur late during the natural history of the illness. Noteworthy, depending on the type of diabetes and on the disease stage, TRAIL can have a dual role, either as immune modulator as well as a regulatory molecule of the vascular wall fitness. PMID- 17465341 TI - If you think you're a liberal, wait awhile. PMID- 17465342 TI - The radiographic bone loss pattern adjacent to immediately placed, immediately loaded implants. AB - PURPOSE: The purpose of this study was to evaluate radiographic bone levels adjacent to implants placed in fresh extraction sockets (ESs) and immediately loaded with a fixed full-arch provisional restoration compared to bone levels adjacent to implants placed in native bone (NB) under the same restorative conditions. MATERIALS AND METHODS: Patients with a hopeless maxillary and/or mandibular dentition had their remaining teeth extracted and 6 to 8 implants placed and restored within 72 hours. Radiographs were obtained at time 0, 3 to 6 months, and annually for 5 years. The radiographs were digitized, and the bone level changes were measured using a computer-assisted method. RESULTS: A total of 139 implants, 42 ES and 97 NB, placed in 17 patients were evaluated. The overall results indicated that for all implants (ES + NB), 0.60+/-0.71 mm of bone was lost after 6 months; 1.17 +/-0.59 mm of bone was lost after 18 months; 0.87+/ 0.76 mm bone was lost after 36 months; and 1.35 +/-0.42 mm of bone was lost after 60 months. When stratifying for NB versus ES implants, it was found that for NB implants, 0.75+/- 0.21 mm of bone was lost after 6 months; 1.31 +/- 0.91 mm of bone was lost after 12 months; 1.07+/-0.21 mm of bone was lost after 36 months; and 1.45 +/- 0.49 mm of bone was lost after 54 months. For ES implants, 0.14 +/ 0.33 mm of bone was lost after 6 months; 1.02 +/-0.27 mm of bone was lost after 12 months; 0.86+/-0.42 mm of bone was lost after 36 months; and 1.30 +/-0.48 mm of bone was lost after 54 months. CONCLUSION: The combination of ES and NB implants can be immediately loaded with a fixed full-arch prosthesis and remain stable for greater than 5 years. The bone loss adjacent to these implants is similar to that seen surrounding those placed and restored using traditional protocols. PMID- 17465343 TI - Long-term results in placement of screw-type implants in the pterygomaxillary pyramidal region. AB - PURPOSE: To present the long-term results for screw-type implants consecutively placed in the pterygomaxillary-pyramidal region in the treatment of patients with posterior maxillary edentulism. MATERIALS AND METHODS: One hundred fifty-two implants were inserted in 92 partially edentulous patients using cylindric osteotomes as bone-site formers, thus minimizing the use of drills in the bone preparation. RESULTS: The 152 implants placed were loaded for an average of 89.7+/-30.7 months. There were a total of 8 failures; 6 took place between implant placement and prosthesis delivery (early failures), and 2 failed in following functional loading. The overall survival rate was 94.7%. DISCUSSION: Cylindric osteotomes allow the surgeon to accurately place implants. The use of surgical drilling was minimized, resulting in lowered surgical risk and diminished bone loss. CONCLUSION: Implant placement in the posterior pterygomaxillary region using cylindric osteotomes for osteotomy preparation resulted in an implant survival rate of 94.7%. PMID- 17465344 TI - Evaluation of autogenous bone grafts, particulate or collected during osteotomy with implant burs: histologic and histomorphometric analysis in rabbits. AB - PURPOSE: The aim of this study was to evaluate bone regeneration in bone cavities filled with particulate autogenous bone either harvest in blocks and subjected to milling procedures or collected during osteotomy with implant burs. MATERIALS AND METHODS: In 12 rabbits, 3 noncritical unicortical cavities 7 mm in diameter were prepared with a trephine drill on the right tibia. The cavities were filled respectively with particulate autogenous bone achieved with a manual bone crusher (particulate group), with particulate autogenous bone obtained using bone collector during osteotomy (collected group), and with blood clot (control group). Animals were sacrificed at 7, 15, and 30 days after surgery (4 animals for each time period). The sections were examined by histologic and histomorphometric analysis. RESULTS: At 7 days, the samples were filled by coagulum, and bone particles were observed only in the collected (24%) and particulate groups (44.75%). At 15 days, there was connective differentiation in all groups, with presence of grafted bone particles and onset of newly formed bone in the collected (38.88%) and particulate groups (46.0%). At 30 days, there was bone fill (immature trabecular bone) of the cavities in the control (50%), collected (64.63%) and particulate groups (66%). CONCLUSION: No significant difference was demonstrated between noncritical unicortical bone defects in rabbit tibiae filled with particulate bone harvested as a block and subjected to milling and those filled with bone collected during osteotomy with implant drills when the defects were observed up to 30 days following their creation. PMID- 17465345 TI - Bone density assessments of dental implant sites: 3. Bone quality evaluation during osteotomy and implant placement. AB - PURPOSE: In previous publications of this series of studies on human cadaver jaws, bone densities were assessed and compared using subjective evaluation, conventional computed tomography (CT), and cone-beam computed tomography (CBCT). The aim of this study was to compare subjective bone quality during osteotomy and implant insertion resistance torque to noninvasive subjective and objective radiographic bone density assessments. MATERIALS AND METHODS: Forty-two designated implant sites were selected. Self-tapping implants were inserted into these sites. The operator subjectively rated the bone density during the osteotomy procedure. Resistance torque was recorded during insertion of the implants. RESULTS: Subjective drilling resistance was modestly correlated to subjective radiographic density evaluation (Lekholm and Zarb; Spearman's rho of 0.53, P < .001). Subjective drilling resistance compared to the bone density in Hounsfield units (HU) obtained using CT and CBCT showed correlation coefficients of 0.61 and 0.59, respectively (P < .001). Significant overlap of density values was found for adjacent drilling ratings. On average, a difference in bone density of 180 HU was required to identify differences between drilling resistance groups. Comparisons of 2 implant insertion resistance torque variables (highest reading and regression slope of available readings) with CT and CBCT HU showed correlation coefficients of 0.61 to 0.63 (P < .01). CONCLUSION: Insertion torque resistance was modestly correlated with objective CT and CBCT measurements of bone density. The merit of these assessments of cadavers awaits clinical study. PMID- 17465346 TI - Analysis of stress and strain around orthodontically loaded implants: an animal study. AB - PURPOSE: The aim of this study was to describe the stress and strain fields around orthodontically loaded dental implants using the finite element method and to evaluate the relationship between the generated strain and the biologic reaction expressed through histomorphometric parameters. Finally, this study aimed to evaluate the interaction between the orthodontic loading and the deformation generated by normal occlusal function. MATERIALS AND METHODS: Sixteen titanium dental implants were inserted in extraction sockets after the removal of the second premolars and first molars of 4 adult Macaca fascicularis monkeys. After 17 weeks of healing, the implants were loaded by a pair of Sentalloy springs (50 cN) for 16 weeks. After sacrifice, tissue blocks including the implants and surrounding bone were excised. Five tissue blocks were scanned with a synchrotron radiation-based microtomography (microCT) scanner and sample specific finite element models were generated. Subsequently all samples were prepared for histomorphometric analysis. RESULTS: All implants were osseointegrated, although the surrounding alveolar bone differed from sample to sample. As a consequence the finite element analyses showed that the stresses and strains in the peri-implant alveolar bone greatly varied among the samples. A high level of remodeling activity was found close to the implants. DISCUSSION: Individual differences between the receptors (in this case, the monkeys) have a large effect on both the biologic and morphologic parameters. These variations were indeed found to have a substantial impact on the (re)modeling dynamics and the load transfer mechanisms around the implants. CONCLUSIONS: By integrating different analysis techniques to evaluate bone (re)modeling around orthodontically loaded implants, this study has demonstrated the complexity and case-specific character of alveolar adaptation to orthodontic loading. Furthermore, stresses generated by combined functional and orthodontic forces should not be neglected. PMID- 17465347 TI - Clinical evaluation of a prospective multicenter study on 1-piece implants. part 1: marginal bone level evaluation after 1 year of follow-up. AB - PURPOSE: The aim of the present investigation was to evaluate the marginal bone level after 1 year of follow-up of 1-piece implants after immediate provisional restoration fabrication. MATERIALS AND METHODS: Patients received NobelDirect and NobelPerfect 1-piece implants (Nobel Biocare, G6teborg, Sweden) that were immediately restored and placed into function as part of a 3-year, multicenter investigation. Life table analysis was used for evaluation of implant cumulative survival rates. The Student t test (for dichotomous variables) and Pearson correlation (for continuous and ordered categorical variables) were used to estimate the influence of separate parameters on marginal bone. RESULTS: Eighty seven patients received 152 NobelDirect or NobelPerfect 1-piece implants. Of these, 81 patients returned for the 1-year follow-up and 21 for the 2-year follow up. Three implants were lost, resulting in a cumulative survival rate of 97.9% after up to 2 years. The average marginal bone level at implant placement was 0.33 mm (SD 1.20, n = 141) superior to the reference point (lower edge of the implant collar). Marginal bone level was -0.77 mm (SD 1.33, n = 138) at 6 months and -0.98 mm (SD 1.38, n = 123) at 1 year. Average bone level at the 2-year follow-up was 0.17 mm (SD 1.20, n = 26). After 1 year of loading, bone level in the maxilla was more apical compared to that in the mandible (P = .05), and a positive correlation was found between bone level at placement and bone level at 12 months (P = .008). Shallow implant positioning resulted in less marginal bone remodeling compared to deep implant positioning. CONCLUSION: On the basis of this prospective multicenter study, stable marginal bone level and soft tissue support the hypothesis that the 1-piece implant has the capacity to preserve both hard and soft tissue. PMID- 17465348 TI - Resonance frequency analysis assessment of implant stability in labial onlay grafted posterior mandibles: a pilot clinical study. AB - PURPOSE: The objectives of this study were (1) to compare the stability, evaluated by means of resonance frequency analysis (RFA), of implants placed posterior mandibles augmented with autogenous bone harvested from the mandibular symphysis with that of implants placed in nongrafted edentulous posterior mandibles and (2) to compare peri-implant marginal bone height changes and implant failure for the 2 groups. MATERIALS AND METHODS: Eight patients with thin posterior mandibular ridges (buccolingual crestal width less than 4 mm) underwent labial onlay alveolar grafting with symphyseal bone blocks 4 months prior to placement of 17 implants. Seven nongrafted patients received 18 implants in the edentulous posterior mandible; these patients served as a control group. RFA was performed the day of implant placement (baseline), 1 month postplacement, 4 months postplacement (after prosthesis delivery), and 12 months postloading. Peri implant bone height changes at a level of 0.01 mm were assessed using periapical radiographs at baseline, the 1-month follow-up, and the 4-month follow-up. Analysis of variance was used to evaluate statistical differences within the groups, and t test was used to make comparisons between groups. RESULTS: None of the patients presented postoperative complications or implant failure. Mean implant stability quotient (ISQ) was 63.0+/-6.0 to 70.2 +/-3.5 for the grafted group and 64.1+/-4.1 ISQ to 70.1+/-3.9 for the nongrafted group. No significant difference was found in mean ISQ between the grafted and nongrafted groups at baseline, the 1-month follow-up, 4 months postplacement, or 12 months postloading (P = .211, P = .873, P = .925, P = .735, respectively). Mean peri-implant bone loss was 0.16 +/- 0.04 mm mesially and 0.16 +/- 0.05 mm distally. CONCLUSION: RFA revealed no difference in implant stability between mandibular ridges augmented with autologous bone grafts at baseline or after loading. PMID- 17465349 TI - Midline mandibular deformation during nonmasticatory functional movements in edentulous subjects with dental implants. AB - PURPOSE: Mandibular deformation during function in patients with implant supported prostheses is associated with increased strain at the bone-implant interface when dental implants are connected by a relatively rigid superstructure. Whilst there is a body of evidence concerning deformation as measured between the corpora, there are little data on its effects in the midline. This study measured 3 types of midline mandibular deformation during nonmasticatory functional mandibular movements in edentulous subjects with dental implants. MATERIALS AND METHODS: A range of custom displacement transducers was fabricated for 5 edentulous subjects who had been treated with dental implants in the anterior mandible. These transducers were mounted on contralateral implant abutments adjacent to the midline to measure medial convergence, corporal rotation, and anteroposterior shear. Their output was recorded for offline analysis by a personal computer. RESULTS: The values of medial convergence ranged from 15 to 42 microm during opening, from 10 to 21 microm during lateral excursions, and from 18 to 53 microm during protrusion. Corporal rotation varied from 0.05 to 0.11 degrees during opening, from 0.03 to 0.08 degrees during lateral excursions, and from 0.03 to 0.15 degrees during protrusion. Anteroposterior shear varied from 38 to 93 pm during opening, from 28 to 56 microm during lateral excursions, and from 52 to 103 pm during protrusion. DISCUSSION AND CONCLUSIONS: Nonmasticatory physiological mandibular movements cause the jaw to deform about the midline in at least 3 directions. It is important for the clinician to be aware of the phenomenon of mandibular deformation, which should be taken into consideration in the design and monitoring of mandibular prostheses. PMID- 17465350 TI - Resistance of internal-connection implant connectors under rotational fatigue loading. AB - PURPOSE: To aid in developing mechanically optimized implant-abutment connectors, the fatigue resistance of 5 connector configurations of the Replace Select system (Easy abutment, Easy abutment without antirotational mechanism, Multi-unit abutment, Esthetic Alumina abutment, Esthetic Zirconia abutment) was investigated. Other purposes of the study were to determine whether the connector's antirotational mechanism participates in fatigue resistance and to compare the results with previous data on Straumann connectors. MATERIALS AND METHODS: The repetitive, alternating, and multivectorial intraoral force pattern was reproduced by subjecting the test specimens to the rotating cantilever beam test. To this end, the samples were spun around their long axis while clamped into a revolving collet on one end and loaded normal to their long axis on the other end. The aim was to determine the load level at which 50% of the specimens survived and 50% fractured before 106 cycles. Means were determined using the staircase procedure. They were fitted with 95% confidence intervals for intergroup comparisons. RESULTS: In the chosen testing configuration, 2 statistical groups emerged. The Easy abutments with and without antirotational mechanism were statistically similar, with mean failure loads in the 70 to 72 N range. Both ceramic and the Multi-unit abutments belonged to the second group, with mean failure loads in the 53 to 58 N range. CONCLUSIONS: (1) The fatigue resistance of ceramic and the Multi-unit abutments was approximately 20% less than that of the Easy Abutments. (2) The antirotational mechanism did not participate in mechanical resistance. (3) The fatigue strength of the Easy abutment connectors was approximately 20% greater than the equivalent abutments in the Straumann system. PMID- 17465351 TI - Bone regeneration using a synthetic matrix containing a parathyroid hormone peptide combined with a grafting material. AB - PURPOSE: The aim of the present study was to test whether a newly developed synthetic matrix made of polyethylene-glycol (PEG) containing a covalently bound peptide of the parathyroid hormone (PTH1-34) enhances bone regeneration compared to grafting procedures and to spontaneous healing. MATERIALS AND METHODS: In each of 16 rabbits used, 4 titanium cylinders were screwed into perforated slits made in the cortical bone of the calvaria. The cylinders were either left empty (control) or filled with 1 of the following: (1) PEG matrix and hydroxyapatite/tricalcium phosphate (HA/TCP) granules, (2) PEG matrix containing 100microg/mL of PTH and HA/TCP granules, or (3) PEG matrix containing 20microg/mL of PTH1-34 and HA/TCP granules. After 8 weeks, the animals were sacrificed, and ground sections were obtained for histology. RESULTS: Quantitative histomorphometry demonstrated a significantly increased amount of newly formed bone for PTH1-34 compared to sites treated with PEG and HA/TCP and to empty control sites (P < .01; analysis of variance and subsequent pairwise Student t test). The mean percentages of mineralized bone were 19.6%+/-6.0% for 100 microg/mL PTH, 18.0% +/- 6.2% for 20microg/mL PTH, 12.0% +/-6.5% for PEG and HA/TCP without PTH, and 10.5% +/-3.7% for the empty control. The mean areas of bone regenerated within the cylinders were 53.5% +/-22.7% for 100 microg/mL PTH, 51.1% +/-22.6% for 20 microg/mL PTH, 34.3% +/- 22.5% for PEG and HA/TCP without PTH, and 23.2% +/-10.1% for the empty control. DISCUSSION: Human and animal trials have demonstrated that daily systemic injection of PTH increases bone mineral density. The present study showed that local administration of PTH was also effective in stimulating bone formation. CONCLUSION: It is concluded that this synthetic PEG hydrogel containing a covalently bound peptide of the PTH combined with HA/TCP granules significantly stimulated in situ bone augmentation in rabbits. PMID- 17465352 TI - A retrospective evaluation of a treatment protocol for dental implant periapical lesions: long-term results of 39 implant apicoectomies. AB - PURPOSE: The goal of this study was to evaluate retrospectively the efficacy of a treatment modality for a lesion at the apical portion of a nonmobile dental implant. MATERIALS AND METHODS: All patients were treated with an intraoral treatment approach. A flap was elevated facial to the effected implant site, exposing the bone. A carbide bur was used to open a window in the bone. The bur was then used to cut the implant and completely remove the affected portion of the implant. The surgical site was then closed with interrupted vicryl sutures, and patients were prescribed. RESULTS: Thirty-nine implants in 35 patients with a mean age of 58.3 years were identified clinically and radiographically with the presence of a periapical lesion. These 39 implants, which constituted 9.9% of implants (39 of 395) placed in these 35 patients, were consecutively treated using the intraoral apicoectomy procedure. Thirty-eight of the 39 implants (97.4%) treated with this technique remained stable and continued in function with no further complication. Follow-up time averaged 4.54 years; the longest follow-up time exceeded 15 years. CONCLUSION: Based upon the results of this retrospective study, lesions in the apical region of implants can be treated successfully using an intraoral apicoectomy procedure. PMID- 17465353 TI - Histomorphometric study of ion implantation and diamond-like carbon as dental implant surface treatments in beagle dogs. AB - PURPOSE: Improvements in the bone-implant interface can provide clinical benefits, such as increasing the amount of bone in contact with the implant and shortening the time required to achieve sufficient bone appositioning to allow early prosthetic loading. The present study describes the results obtained with 2 new surface treatments: (a) CO ion implantation; and (b) diamond-like carbon (DLC) coating. MATERIALS AND METHODS: Each group (ion implantation, DLC, and the control group, turned titanium) consisted of 12 samples. Beagle dogs subjected to previous partial edentulation were used. Dual histologic evaluation was made of percentage bone-implant contact (% BIC) of all samples based on conventional histomorphometric analysis and environmental scanning electron microscopy (ESEM). RESULTS: The results obtained after 3 and 6 months of dental implant placement showed greater and faster bone integration in the CO ion implantation group (61% and 62% BIC, respectively) compared with the DLC group (47% and 50%); the data corresponding to the ion implanted samples were statistically significant compared with the control group (33% and 49% BIC after 3 and 6 months, respectively). CONCLUSIONS: The results showed improved % BIC for implants with ion-implanted surfaces in comparison to DLC coating and machined controls. Furthermore, bone integration appeared to be accelerated in the ion implantation group, since high % BIC values were recorded in the early stages after in vivo implantation. PMID- 17465354 TI - Augmentation of the posterior atrophic edentulous maxilla with implants placed in the ulna: a prospective single-blind controlled clinical trial. AB - PURPOSE: To evaluate a new method to treat the posterior atrophic edentulous maxilla: dental implants placed in the ulna and transplanted with their surrounding bone blocks as inlays into the sinus. Conventional sinus augmentation with particulated autogenous bone grafts served as a control procedure. MATERIALS AND METHODS: Fifty-two implants were placed in the ulnas of 20 patients. After 6 weeks, bone blocks containing 1 to 3 implants were harvested and transplanted into the sinuses protruding 3 to 4 mm. Implants were left to heal for 6 weeks. Twenty patients with similar treatment indications treated with particulated bone grafts from the mental symphysis, tibia, or iliac crest acted as controls. Grafts were allowed to heal for 6 months in the control group. Fifty-two control-group implants were allowed to heal for 4 months. The main outcome measures were prosthetic and implant success. Stability of individual implants was assessed with Osstell and Periotest at baseline and after 6 and 12 months of loading. Independent sample chi-square tests, t tests, and paired t tests were used with a significance level of .05. RESULTS: No patient dropped out or withdrew; no prosthesis or implant failed. No major surgical complications were occurred. There were no differences between the 2 groups at any time point in implant stability. Both modalities resulted in a significant increase of implant stability at 6 and 12 months. The mean change (SD) from baseline to 1 year in Periotest measurements was 1.44 (0.48) in the test and 1.29 (0.58) in the control (paired t tests; P < .001). For the Osstell, these values were -5.88 (4.18) and 5.48 (3.93) for the test and control groups, respectively (paired t tests: P < .001). CONCLUSIONS: Ulna implant block grafting represents an alternative to conventional sinus augmentation, particularly when vertical augmentation is desirable or large iliac crest grafts are needed. PMID- 17465355 TI - Efficacy of clinical methods to assess jawbone tissue prior to and during endosseous dental implant placement: a systematic literature review. AB - PURPOSE: To evaluate the evidence for the diagnostic accuracy of clinical methods to assess bone density, quantity, or quality prior to and during dental implant placement. MATERIALS AND METHODS: A PubMed literature search with specific indexing terms and a hand search were made. From the retrieved titles and abstracts, 3 reviewers selected publications on the basis of predetermined inclusion and exclusion criteria. Data were extracted from the selected publications using a protocol. Original studies were interpreted with the aid of the Quality Assessment of Diagnostic Accuracy Studies (QUADAS) tool. RESULTS: The literature search yielded 145 titles and abstracts, of which 71 were selected and read in full text. Seven studies were judged relevant. In 1 study, no method was presented as the test method. In 6 studies, the results of the test method were compared to those of a reference method. However, only 1 study presented the results in terms of percentage of correct diagnoses. In that study, the use of periapical radiography together with reference images yielded correct assessment of the trabecular pattern of the mandible in 3 categories in 58% of the sites. The kappa index of interobserver agreement was 0.35 and intraobserver agreement was 0.67. Corresponding kappa values for 4 classes of bone quality presented by Lekholm and Zarb were 0.33 and 0.43, respectively. No study examined the accuracy of the method originally described by Lekholm and Zarb. CONCLUSION: The evidence for the efficacy of clinical methods to assess jawbone tissue prior to and during endosseous dental implant placement is sparse. This emphasizes the need for studies that incorporate accepted methodologic criteria for diagnostic efficacy. PMID- 17465356 TI - Influence of off-axis loading of an anterior maxillary implant: a 3-dimensional finite element analysis. AB - PURPOSE: To evaluate the influence of the stress/strain distribution in bone around an anterior maxillary implant using 2 types of bone and under 3 different loads. MATERIALS AND METHODS: A premaxillary finite element model featuring an implant and its superstructure was created. Six different testing conditions incorporating 2 types of cancellous bone (high density and low density) under 3 different loading angles (0, 30, and 60 degrees) relative to the long axis of the implant were applied in order to investigate resultant stress/strain distribution. RESULTS: The maximum equivalent stress/strain increased linearly with the increase of loading angle. For each 30-degree increase in loading angle, the maximum equivalent stress in cortical bone increased, on average, 3 to 4 times compared with that of the applied axial load. In addition to loading angle, bone quality also influenced resultant stress distribution. For the low-density bone model, a substantial strain in the cancellous bone was found not only near the implant neck but also at the implant apex. CONCLUSION: To achieve a favorable prognosis under off-axis loading of an anterior maxillary implant, careful case selection for appropriate bone quality and precise occlusal adjustment should be attempted to optimally direct occlusal force toward the long axis of the implant. PMID- 17465357 TI - Hearing preservation surgery for neurofibromatosis Type 2-related vestibular schwannoma in pediatric patients. AB - OBJECT: The authors reviewed the proportion of pediatric patients with neurofibromatosis Type 2 (NF2) in whom hearing was preserved after middle fossa resection of vestibular schwannoma (VS). METHODS: In this retrospective chart review the authors examined the cases of 35 children with NF2 who had undergone middle fossa resection (47 surgeries) between 1992 and 2004 in a neurotological tertiary care center. Surgical outcome was assessed using pure-tone average (PTA) thresholds obtained before and immediately after resection. Speech discrimination scores (SDSs) and pre- and postfacial nerve grades were also recorded. In 55% of surgeries, hearing of less than or equal to 70 dB PTA was maintained postoperatively. The American Academy of Otolaryngology-Head and Neck Surgery Class A hearing (PTA < or = 30 dB and SDS > or = 70%) was preserved in 47.7%. Facial nerve function was good (House-Brackmann Grades I or II) in 81% of the patients. Twelve patients had bilateral middle fossa resections; in nine (75%) of these patients hearing was maintained postoperatively in both ears. CONCLUSIONS: More than half of the children with NF2 in the authors' cohort experienced hearing preservation after middle fossa resection was performed for VS. The authors recommend this approach for preserving hearing in children with NF2. PMID- 17465358 TI - Resection of myxopapillary ependymomas in children. AB - OBJECT: Currently, the optimal treatment of children harboring myxopapillary ependymomas of the spinal cord remains somewhat debatable. The authors present a retrospective study in which they evaluated the records of patients in whom resection of these lesions had been performed. METHODS: Fourteen pediatric patients who had undergone resection of a spinal cord tumor between September 1982 and July 2004 were identified from the database as having histologically classified myxopapillary ependymomas. There were 10 boys and four girls ranging in age from 7 to 18 years (mean age 12.6 years); 71% of the patients were boys. The clinical presentation of the tumor's course was slow and indolent, and the patients had a mean symptom duration of 19.6 months. Twelve patients, who underwent a total of 16 operations, were available for long-term follow-up review. Thirteen gross-total resections and three subtotal resections were performed. There were no deaths due to surgery. Postoperatively, patients initially remained at their preoperative level of function or improved. Patients who had undergone previous surgery and radiotherapy were treated more conservatively than patients who were undergoing surgery for the first time. Four children experienced significant complications following treatment. CONCLUSIONS: As the authors demonstrate in this study, excellent outcomes may be obtained with the use of aggressive surgical techniques with the goal being that of gross-total resection. Despite the best of resections, however, the risk of recurrence remains. Therefore, periodic neuroimaging surveillance of the neuraxis and close clinical follow up are warranted throughout the patient's life. The role for adjunctive chemo- and radiotherapy remains to be defined in the management of myxopapillary ependymomas. PMID- 17465359 TI - Outcome following decompressive craniectomy in children with severe traumatic brain injury: a 10-year single-center experience with long-term follow up. AB - OBJECT: The authors examine the indications for and outcomes following decompressive craniectomy in a single-center pediatric patient population with traumatic brain injury (TBI). METHODS: A retrospective review of data was performed using a prospectively acquired database of patients who underwent decompressive craniectomy at the authors' institution between January 1995 and April 2006. The patients' neuroimages were examined to evaluate the extent of intracranial injury, and the patients' records were reviewed to determine the admission Glasgow Coma Scale (GCS) score, the extent of systemic injuries, the time to craniectomy, and the indications for craniectomy. Long-term functional outcome and independence levels were evaluated using the Glasgow Outcome Scale (GOS) and a Likert patient quality-of-life rating scale. Twenty-three craniectomies were performed in children during the study period. The mean patient age at craniectomy was 11.9 years (range 2-19 years). In all patients, the computed tomography scans obtained at presentation revealed pathological findings, with diffuse axonal injury and traumatic contusions being the most common abnormalities. The median presenting GCS score was 4.6 (range 3-9). Nineteen patients (83%) suffered from other systemic injuries. One patient (4%) died intraoperatively and six patients (26%) died postoperatively. Postoperative intracranial pressure (ICP) control was obtained in 19 patients (83%); an ICP greater than 20 mm Hg was found to have the strongest correlation with subsequent brain death (p = 0.001). The mean follow-up duration was 63 months (range 11-126 months, median 49 months). The mean GOS score at the 2-year follow-up examination was 4.2 (median 5). At the most recent follow-up examination, 13 (81%) of 16 survivors had returned to school and only three survivors (18%) were dependent on caregivers. CONCLUSIONS: Although the mortality rate for children with severe TBI remains high, decompressive craniectomy is effective in reducing ICP and is associated with good outcomes in surviving patients. PMID- 17465360 TI - Long-term motor outcome analysis using a motor score composite following surgical brachial plexus repair. AB - OBJECT: Due to the complex and variable nature of brachial plexus injury, outcome analysis can be cumbersome and imprecise. Many scales have been devised, but no single scale is used uniformly. Moreover, despite several studies in which the authors have reported brachial plexus surgical data, no highly predictive clinical model has been defined. METHODS: In this study the authors performed a retrospective analysis of 114 consecutive brachial plexus surgeries performed by the senior author during the past 14 years at St. Louis Children's Hospital. Of these, 63 are included in this study. The authors defined the motor score composite (MSC) and used this novel metric to perform a detailed analysis of their surgical outcomes. The mean MSC was 0.50 preoperatively, 0.71 at 1 year postoperatively, and 0.80 at 2 years postoperatively. By 2 years postoperatively, 89% of the patients attained a good or excellent recovery. Age at surgery, time to visit, location, and severity were predictive of outcome. Using MSC data, the authors developed a prognostic model that enabled the prediction (with 88% accuracy) of surgical outcomes using preoperative variables. CONCLUSIONS: The MSC is an efficient metric for the reporting of data regarding outcomes of brachial plexus injury. It provides information about extent and severity of injury in a single proportion and facilitates complex data analysis. The authors used the MSC model to accurately predict surgical outcome. This metric could have wide applicability for the prediction of postoperative recovery to improve both surgical decision making and family counseling. PMID- 17465361 TI - Predicting functional change from preintervention measures in selective dorsal rhizotomy. AB - OBJECT: In this investigation the authors attempted to predict change in function following selective dorsal rhizotomy (SDR) and intensive physical therapy in patients with spastic diplegic cerebral palsy (CP) based on multidomain preintervention measures. METHODS: Data pertaining to 22 children with CP were collected before the SDR and again 20 months afterward. Although equations for predicting change in gait speed and function (such as the Gross Motor Function Measure) were derived, the 95% confidence interval (CI) widths were too broad to make accurate predictions that were clinically useful outside the study group. CONCLUSIONS: Future work should be focused on developing additional measures such as lower-extremity motor control and balance in an attempt to reduce the CIs to more clinically relevant values. PMID- 17465362 TI - Effect of shunt catheter on the systemic immune response: evaluation of neutrophil count, function, and rate of chemotaxis. AB - OBJECT: The localized impairment of the host defense mechanism due to the presence of a shunt apparatus has been suggested as a risk factor for shunt infection. The purpose of this study was to evaluate the probable systemic effect of a shunt catheter on neutrophil phagocytosis and chemotaxis in vivo. METHODS: Twenty-four children with hydrocephalus who were referred to the Children's Hospital Medical Center in Tehran for ventriculoperitoneal shunt placement were included in this study. Neutrophil count, chemotaxis, and nitroblue tetrazolium (NBT) tests were performed before and 2 months after the operation. In comparing the preoperative neutrophil count, NBT percentage, and chemotaxis (with and without the addition of a chemoattractant factor) with these same factors postoperatively, the authors found no statistically significant differences. In four children, shunt infections developed during the follow-up period. There were no significant differences between the aforementioned parameters in children with infected shunts and those with uninfected shunts. CONCLUSIONS: The results of this study do not support the idea of systemic impairment of neutrophils after shunt insertion. Further studies with more specific methods are required to elaborate on this issue. PMID- 17465363 TI - Macrocephaly-cutis marmorata telangiectatica congenita. PMID- 17465364 TI - Surgical management of cerebellar tonsillar herniation in three patients with macrocephaly-cutis marmorata telangiectatica congenita. Report of three cases. AB - The authors report on three patients with a congenital brain overgrowth syndrome, macrocephaly-cutis marmorata telangiectatica congenita (M-CMTC), who experienced neurological sequelae associated with herniation of the cerebellar tonsils through the foramen magnum. In two of these patients, the authors document postnatal brain overgrowth that contributed to recurrent descent of the cerebellar tonsils and complicated the surgical treatment in one of the patients. The authors address the neurosurgical concerns related to this syndrome with special attention to acquired tonsillar ectopia and postulate that some patients may be at risk for progressive tonsillar herniation and consequent neurological symptoms due to cerebellar overgrowth. Ectopic cerebellar tonsils have been reported previously in cases of M-CMTC, and this phenomenon may be a secondary event associated with brain overgrowth rather than due to a congenitally small posterior fossa. PMID- 17465365 TI - Hydrogel-induced cervicomedullary compression after posterior fossa decompression for Chiari malformation. Case report. AB - The use of an absorbable hydrogel dural sealant has been approved for neurosurgical applications with no published reports of complications to date. The authors present the case of a 13-year-old girl with syringomyelia and quadriparesis who underwent posterior fossa decompression and dural augmentation for Chiari malformation Type I. Dural closure was performed with a dural substitute patch, hydrogel dural sealant, and gelatin sponge. Magnetic resonance imaging, performed after initial postoperative improvement in the patient's quadriparesis deteriorated, demonstrated an expanding epidural mass collection causing cervicomedullary compression. Exploration on postoperative Day 15 revealed an expanded layer of hydrogel underlying a layer of gelatin sponge. The authors conclude that hydrogel dural sealants should be used cautiously in spaces that cannot tolerate significant mass effect. PMID- 17465366 TI - Mature teratoma associated with an interparietal encephalocele. Case report. AB - Teratomas and encephaloceles are both uncommon congenital malformations that originate near the midline in the head and neck. An encephalocele is an extracranial extension of intracranial structures through a defect in the skull, whereas a teratoma consists of tissue from all three embryonic germ layers and is a true congenital tumor. In the present study the authors describe the first reported case of a mature teratoma associated with an interparietal encephalocele presenting as a large midline subcutaneous mass in a newborn child. The appearance of these two pathological entities together suggests a common origin. The embryological significance of this finding is discussed, and the pertinent literature is reviewed. PMID- 17465367 TI - Primary atypical teratoid/rhabdoid tumor of the clival region. Case report. AB - An atypical teratoid/rhabdoid tumor of the central nervous system (CNS) is a rare, aggressive neoplasm found in infants and children that has similar characteristics to CNS primitive neuroectodermal tumors/medulloblastomas. The authors present the case of a patient with an atypical teratoid/rhabdoid tumor and discuss the imaging, histopathological, immunohistochemical, and cytogenetic findings. Tumor cells displayed positive reactions for vimentin, epithelial membrane antigen, and cytokeratin, and they displayed no reaction for glial fibrillary acidic protein, desmin, and actin. The karyotype was 46, XY. The phenotype of an atypical teratoid/rhabdoid tumor appears heterogeneous when examined by histological, immunohistochemical, and genetic analysis. The authors describe the case of a 4-year-old boy who harbored an atypical teratoid/rhabdoid tumor in the clivus, which appeared as a chordoma on neuroimages. To their knowledge, this location of an atypical teratoid/rhabdoid tumor has not been described in the literature. PMID- 17465368 TI - Radiosurgical posterior corpus callosotomy in a child with Lennox-Gastaut syndrome. Case report. AB - The authors report the successful use of radiosurgery in a child for posterior corpus callosotomy; the early results are good and the patient has not suffered any morbid conditions. The relevant literature pertaining to the use of radiosurgery for treating epilepsy is reviewed. Details of the radiosurgical techniques and prescription dose used are presented, along with 1-year serial neuroimaging results. PMID- 17465369 TI - Double spinal dysraphism. Report of three cases. AB - The simultaneous presence of multiple spinal neural tube defects is unusual. There have been only a few of these cases reported in the literature. The authors report on three cases of double spina bifida cystica. One patient had two myelomeningoceles (MMCs) at the cervical and lumbosacral regions, one was noted to have both thoracolumbar and sacral defects, and the third presented with double MMCs at lumbar and lumbosacral levels. All three neonates in these cases underwent surgical treatment and ventriculoperitoneal (VP) shunt insertion for associated hydrocephalus. One child died at the age of 2 months despite a well functioning VP shunt. The other two patients had no complications. Current models of neural tube closure do not thoroughly explain the mechanisms of multiple spinal dysraphism, but the multisite closure model provides a better understanding of caudal neural tube closure than other closure-site models. PMID- 17465370 TI - Endovascular placement of a ventriculoatrial shunt. Technical note. AB - Atrial shunt revision surgeries are sometimes difficult due to venous occlusion and neck scarring. A direct approach guided by venography facilitates exposure and guarantees accurate placement of the distal catheter. Five patients with complicated histories of shunt malfunction were treated using an endoscope assisted technique. The distal end of an atrial catheter was advanced into the atrium after having been connected to a venous catheter of a slightly smaller diameter than the one previously advanced from the femoral vein through the atrium. Once the position of the atrial catheter was confirmed fluoroscopically, the venous catheter was detached and removed. No complications developed in any patient. This endoscope-assisted technique offers three advantages: it demonstrates the patency of the jugular vein through venography, facilitates identification of the internal jugular vein in the neck, and provides a quick way to confirm that the distal end of the atrial catheter has been placed correctly. This technique should be considered for use in patients with a history of failed atrial shunts. PMID- 17465371 TI - Iatrogenic intradural epidermoid cyst after lumbar puncture. Case illustration. PMID- 17465372 TI - Intracranial foreign bodies causing delayed brain abscesses: intracranial sewing needles. Case illustration. PMID- 17465373 TI - The Section on Neurological Surgery of the American Academy of Pediatrics: Annual Report 2005 to 2006. PMID- 17465374 TI - Pediatric aneurysms. PMID- 17465375 TI - Chiari malformation. PMID- 17465376 TI - Trapped IV ventricle. PMID- 17465377 TI - [The treatment of acute recurrent pharyngotonsillitis]. AB - AIM OF THE STUDY: In paediatric patients, recurrent febrile pharyngo-tonsillitis is usually due to chronic inflammation of the tonsils and/or adenoids: these episodes are often associated with other clinical manifestations (respiratory obstruction of naso-pharyngeal origin; auricular inflammation, especially effusive otitis media and acute otitis media; streptococcal beta-haemolytic Group A (SBEGA) infection, causing a distant disorder of varying severity and which may, therefore, give rise to serious pathological conditions). A retrospective multicentric investigation has, therefore, been carried out in order not only to further elaborate findings emerging from earlier studies, performed in other countries, in which random methods were used, but also to focus on: 1. the possible association of these clinical manifestations with recurrent febrile pharyngo-tonsillar inflammation in relation to frequency of the condition; 2. results obtained with adenoidectomy or with adeno-tonsillectomy, referring to the overall clinical outcome of the pathological condition, bearing in mind data emerging from the control groups, submitted to strict follow-up, associated with any medical treatment deemed necessary. MATERIAL AND METHODS: The study population comprised 692 patients (394 male, 298 female), age range 2-11 years and with a minimum follow-up of 2 years. Considering the clinical manifestations emerging from the case history, the following two investigations were carried out: 1. the first, on 501 patients, all previously submitted to adeno tonsillectomy in whom the recurrent febrile pharyngo-tonsillar inflammation had occurred, respectively > 4 or < or = 4 episodes during the last 12 months; 2. the second, on 455 patients in whom recurrent febrile pharyngo-tonsillar inflammation, in the 12 months prior to the beginning of the study, had been < or = 4 episodes: in 264 cases, adeno-tonsillectomy was carried out; in 144, adenoidectomy; 47 children were studied as controls and, therefore, submitted to watchful waiting. The results obtained were statistically analysed by chi2 and Fisher test: a "p" value of < 0.05 was considered statistically significant. RESULTS: The first investigation showed that all symptomatological manifestations considered were resolved following adeno-tonsillectomy, in a significantly high percentage of cases (recurrent febrile pharyngo-tonsillar in flammation: 91.8%; naso-pharyngeal respiratory obstruction: 92.2%; pathological ASO titre: 69.8%; distant disorders probably due to SBEGA: 76%; effusive otitis media: 76%; acute recurrent otitis media: 87.5%). The most interesting clinical findings obtained can be summarized as follows: naso-pharyngeal respiratory obstruction is a frequent finding; the incidence is statistically higher in cases in which recurrent febrile pharyngo-tonsillar inflammation is less frequent (p < 0.05): in these patients, this is an important pathological manifestation indicating the need for surgery; the higher incidence is associated with at significant finding of pathological auricular processes; the percentages of recovery for naso pharyngeal respiratory obstruction are statistically greater in the group of patients in whom the recurrent febrile pharyngo-tonsillar inflammation is more frequent: an identical result is found in patients with the effusive form of otitis media and in those with recurrent acute otitis media; a pathological ASO titre is more frequently found, the difference being statistically significant (p < 0.05), in the group of patients with more frequent recurrent febrile pharyngo tonsillar inflammation, albeit this factor does not affect the percentage of recovery; also the favourable evolution and recovery of clinical distant manifestations, related to streptococcal beta-haemolytic Group A infection, are correlated with the return to normal of the ASO titre. The second investigation demonstrated the efficacy of surgical treatment, adeno-tonsillectomy and adenoidectomy, in all the clinical manifestations studied, the possibility of achieving resolution of recurrent febrile pharyngo-tonsillar inflammation even in the control subjects submitted to medical treatment, but also the lack of success, in the latter, as far as concerns naso-pharygeal respiratory obstruction, the pathological ASO titre and the inflammatory auricular processes. CONCLUSIONS: The present investigations confirm previous observations emerging from randomized clinical studies, regarding the possibility to achieve valid prevention of recurrent febrile pharyngo-tonsillar inflammation, also in a large percentage of control patients submitted to watchful waiting; these investigations, however, also demonstrated that, in the control group, the percentage of subjects in whom recovery of other symptoms was achieved was very low compared to that in the groups submitted to surgery. In other words, clearly emerges as far as concerns the overall symptomatological picture of the patients, the efficacy of the surgical treatment, but also the lack of success of the medical treatments in all clinical manifestations associated with recurrent febrile inflammations. Therefore, it is necessary to select and advise the most suitable treatment strategy, not in relation to the Guidelines elaborated on the basis of results emerging from statistical research, that, due to the intrinsic methodology limits cannot be applied to all cases in an acritical fashion, but in relationship to the specific clinical picture of each individual patient. PMID- 17465378 TI - [Acute recurrent pharyngotonsillitis and otitis media]. AB - BACKGROUND: In the more recent randomized clinical studies, the surgical procedures adopted (adenoidectomy and adeno-tonsillectomy) for the treatment of the effusive and the acute recurrent forms of otitis media have generally shown a relatively modest and not persistent efficacy. It should be pointed out, however, that the cases studied displayed no clinical elements suggesting the existence of any aetio-pathogenic relationship between the auricular inflammation and a possible pathological disorder localized in the tonsils and/or adenoids. AIM OF THE STUDY: The present investigation aimed to establish: a) the frequency with which the two forms of tympanic inflammation, under study, were detected in children with recurrent febrile pharyngo-tonsillar inflammation, also considering the possible presence of naso-pharyngeal respiratory obstruction and the severity of these two pathological manifestations; b) in relationship to these parameters, what might be the outcome with respect to the pathological auricular processes following surgical treatment of the pharyngeal disorder (adeno-tonsillectomy or adenoidectomy) vs. watchful waiting, c) the possible advantages offered by ventilation tube, inserted during surgical treatment, in the prevention of the two forms of otitis media studied. MATERIAL AND METHODS: This retrospective multi centre investigation involved 678 subjects (388 male, 290 female), age range 2 - 11 years: all subjects were followed-up for a minimum period of 2 years. Patients in this study population were submitted to 3 groups of investigations: a) in the first (487 children submitted to adeno-tonsillectomy), the study population was subdivided into 2 groups according to the frequency of the complaints of febrile pharyngo-tonsillar inflammation reported during the previous 12 months (> 4 or < or = 4); b) in the second (443 children with recurrent febrile pharyngo-tonsillar inflammation during the previous 12 months < or = 4), three groups of cases were compared; in the 1st (252 cases), patients were submitted to adeno-tonsillectomy; in the 2nd (144 subjects), adenoidectomy was carried out; in the 3rd (47 cases), the watchful waiting approach was adopted; c) in the third (193 cases, with both pathological auricular manifestations), patients were submitted to adeno tonsillectomy (119 cases) or to adenoidectomy 74 cases); during surgery, ventilation tube was applied bilaterally in 61 cases, while 132 patients formed the control group. The results obtained were statistically analysed by chi2 and Fisher test: a "p" value of < 0.05 was considered statistically significant. RESULTS: The first investigation showed: a) the incidence of cases with auricular disorders was greater in the group presenting recurrent febrile pharyngo tonsillar inflammation < or = 4 (effusive otitis media: p < 0.05); in this group, the presence of cases with naso-pharyngeal respiratory obstruction was statistically greater (p < 0.05); b) the number of cases in which the pathological auricular process was resolved following adeno-tonsillectomy, appeared high (effusive otitis media: 136/179 = 76%; acute recurrent otitis media: 189/216 = 87.5%), not unlike the percentage of patients with positive results on recurrent febrile pharyngo-tonsillar inflammation and on naso pharyngeal respiratory obstruction (91.6% and 92.4%, respectively); c) the percentages of recovery, for auricular disorders, were significantly higher in the group presenting more frequent recurrent febrile pharyngo-tonsillar inflammation, i.e. > 4 (p < 0.05); in this group, also the incidence of cases, in which the naso-pharyngeal respiratory obstruction was resolved, was greater. The second investigation demonstrated, in the groups in which surgical treatment was carried out, compared to controls: a) a significantly greater percentage of cases achieving recovery, for both parameters (p < 0.05); as far as concerns the effusive form of otitis media, a total of 72 (67.3%) subjects were cured following adeno-tonsillectomy; 67 (81.7%), following adenoidectomy; in acute recurrent otitis media, the subjects achieving recovery were; for adeno tonsillectomy, 97 (82.2%); for adenoidectomy; 85 (81.7%); b) in the subjects in whom, following surgical treatment, the inflammatory auricular disorder was resolved, it was generally found that recovery had been achieved not only for the recurrent febrile pharyngo-tonsillar inflammation but also for the naso pharyngeal respiratory obstruction; c) in the controls, the results appeared to be significantly less satisfactory (< 0.001); together with the lack of success in the watchful waiting approach, as far as concerns auricular disorders, we found, likewise, a significant lack of efficacy (p < 0.001) of this treatment in the management of naso-pharyngeal respiratory obstruction (5/35 cases cured: 14.3%). The third investigation showed that the application of the ventilation tube during adeno-tonsillectomy or adenoidectomy did not lead to any improvement in the results; indeed, the percent recovery of the pathological tympanic processes was significantly greater (p < 0.05) in those subjects in whom transtympanic drainage had not been adopted. CONCLUSIONS: The results of these investigations reveal a clear relationship between the naso-pharyngeal respiratory obstruction and the pathological auricular processes in subjects with episodes of recurrent febrile pharyngo-tonsillar inflammation. These results also demonstrate that adeno-tonsillectomy and/or adenoidectomy are able to resolve the auricular inflammatory manifestations in a very large percentage of cases. These data are apparently in contrast with those described in recent reports related to randomised investigations. In effect, the two groups of investigations had different aims: the studies carried out according to a randomised approach were performed on study populations in which the findings of pathological auricular manifestations were not linked by the demonstration of a correlation with possible adenoid or adeno-tonsillar inflammation; the present study, on the other hand, was performed on patients in whom the auricular disorder was part of the clinical picture of recurrent febrile pharyngo-tonsillar inflammation. Therefore we believe that each of these study methods has precise possibilities, but also limitations; thus, one should not exclude the other, but rather be integrated. PMID- 17465379 TI - Treatment of subdural hematomas in infants. PMID- 17465380 TI - Factors influencing the complication rate of subduroperitoneal shunt placement for the treatment of subdural hematomas in infants. AB - OBJECT: In this study the authors' goal was to identify the complication rate of subduroperitoneal (SDP) shunts for the treatment of subdural hematomas (SDHs) in infants and to determine the influences on and predictive factors for these complications. METHODS: The authors present a case series spanning the years 1994 to 2003 and include a statistical analysis of 161 children 2 years of age or younger with SDH who were treated using a unilateral valveless SDP shunt. The patient history, characteristics, and treatment methods including prior therapies, neuroimaging findings, and clinical outcomes were measures of evaluation. Thirty-six children (22.4%) suffered complications related to SDP shunts: obstruction in 27 (16.8%), infections in eight (5%), disconnection in four (2.5%), migration in three (1.9%), wound complications (leakage and skin ulceration) in two (1.2%), and symptomatic subdural rebleeding in one (0.6%) necessitating bur hole evacuation. Seventeen children (10.6%) underwent placement of a second SDP shunt because of ipsilateral or contralateral persistent fluid collections, or premature shunt removal. With the exception of 12 patients (7.4%), shunt removal was performed systematically and resulted in the following minor complications in 30 children (18.6%): an adherent proximal catheter in 16 (9.9%), transient symptoms of intracranial hypertension in six (3.7%), subcutaneous cerebrospinal fluid accumulation in four (2.5%), local infections in three (1.9%), and hydrocephalus requiring placement of a ventriculoperitoneal shunt in one (0.6%). Status epilepticus at presentation and neuroimaging findings such as areas of hyperdensity on computed tomography (CT) scans representing fresh blood in the subdural fluid collections before shunt insertion and at follow up were predictors of shunt-related complications. Correlations were also discovered for the following CT findings: ischemic lesions before shunt treatment, cerebral atrophy and ventricular dilation during the last follow up, and residual medium to large collections before shunt removal. Children who attained a good outcome were less affected by shunt-related complications, unlike those who presented with focal deficits and/or visual impairment. CONCLUSIONS: Subduroperitoneal shunt placement for the treatment of SDH in infants is--despite the complication rate--an effective and often inevitable treatment option, especially for most large and symptomatic SDHs; a certain number of complications could be reduced with careful and precise surgical techniques. Close observation for detection of risks is mandatory, and seizure control is essential to prevent further brain injury that may result in large subdural fluid collections that are difficult to treat. PMID- 17465381 TI - Pediatric gravitational shunts: initial results from a prospective study. AB - OBJECT: The authors' goal in this paper was to evaluate prospectively the efficacy and safety of a new pediatric gravitational shunt to determine whether it warrants inclusion in a randomized, controlled trial with other shunts. METHODS: A total of 55 children between the ages of 0 and 6 years (median age 0.5 years, average age 4+/-6 years) underwent primary shunt implantation; all received the Miethke Paedi-GAV. The follow-up period ranged between 12 and 77 months (mean 47+/-21 months). The primary end point of the study was the first shunt failure necessitating revision. The 1- and 2-year shunt survival rates were 75 and 68%, respectively. The average failure-free shunt survival duration was 1423 +/- 641 days. Based on imaging findings, no slitlike ventricles occurred. The complication rate was 33%, and the median time to shunt failure was 45 days. Underdrainage occurred in one child (1.8%) and overdrainage in two children (3.6%). CONCLUSIONS: These preliminary results prove the Miethke Paedi-GAV to be a safe and effective pediatric shunt worthy of inclusion in a randomized comparison with other shunts in the pediatric population. PMID- 17465382 TI - Chiari malformation Type I. PMID- 17465383 TI - The role of limited posterior fossa craniectomy in the surgical treatment of Chiari malformation Type I: experience with a pediatric series. AB - OBJECT: Despite progress in the understanding of the causes and pathophysiology of Chiari malformation Type I (CM-I), definitive surgical treatment remains unresolved. Various techniques have been propounded over the years, but there is no general consensus on the most appropriate surgical management for this condition. The authors report their experience with the surgical treatment of 30 pediatric patients with CM-I. METHODS: The results obtained in 30 patients who underwent surgery for symptomatic CM-I with a purely extradural procedure were retrospectively reviewed. The patient age at operation ranged from 2 months to 16 years (mean 68 months). In 26 patients the CM-I occurred as an isolated disease, whereas in four a recognized bone anomaly involving the cranial base (achondroplasia) was observed. All patients underwent suboccipital craniectomy, which was accompanied by C-1 laminectomy in 22. In all cases a thick fibrous band at the level of foramen magnum was resected; in 11 children serial incisions of the outer layer of the dura mater were performed as well to expand the posterior fossa volume. The follow-up duration varied from a minimum of 12 months to 12.6 years (mean 4.7 years). The most frequent symptoms and signs were head and/or neck pain (56.7%), followed by vertigo (27.7%), upper-and lower-extremity weakness (20.0%), and ataxia (20.0%). Syringomyelia was noted in 12 patients (40.0%) on magnetic resonance (MR) images. Improvement in or resolution of clinical symptoms and signs was observed in all patients. However, only minimal or no modifications could be found in the position of cerebellar tonsils on postoperative MR images in most patients (only occasional improvement in tonsillar herniation), whereas syringomyelic cavitations reduced in size in half of the cases. Nevertheless, in only two patients was a reoperation necessary. CONCLUSIONS: In the authors' experience, suboccipital craniectomy and C-I laminectomy (eventually integrated by dural delamination) can represent an effective treatment for symptoms associated with CM-I. PMID- 17465384 TI - Critical analysis of the Chiari malformation Type I found in children with lipomyelomeningocele. AB - OBJECT: Occasional comments are found in the literature regarding patients with lipomyelomeningocele and concomitant Chiari malformation Type I (CM-I). The object of this study was to explore the association between these two conditions. METHODS: The authors performed a retrospective database analysis of lipomyelomeningocele cases to identify cases of concomitant CM-I. Analysis of posterior fossa volume (based on the Cavalieri principle) was performed in all identified cases in which appropriate neuroimages were available, and the results were compared with those obtained in age-matched controls. Seven (13%) of 54 patients with lipomyelomeningocele were found to also have CM-I. Two of these were symptomatic (cervicothoracic syrinx and occipital headaches) and required posterior fossa decompression. No correlation was found between the amount of hindbrain herniation and the level of the conus medullaris or the type of lipomyelome-ningocele (for example, caudal or transitional). Volumetric studies of the posterior fossa revealed normal age-matched volumes in all but one patient (who had asymptomatic CM-I). CONCLUSIONS: The incidence of CM-I in patients with lipomyelomeningocele appears to be significantly greater than that of the general population and the association rate is too high for the finding to be a chance occurrence. Decreases in the volume of the posterior cranial fossa were not found in the majority of patients in this small cohort; therefore, the cause of the concomitant occurrence of lipomyelomeningocele and CM-I remains undetermined. Clinicians should consider obtaining imaging studies of the entire neuraxis in patients with lipomyelomeningoceles and should investigate other causes for syringes found in association with lipomyelomeningoceles. PMID- 17465385 TI - Tethered spinal cord and VACTERL association. AB - OBJECT: Vertebral defects, anal atresia, cardiovascular anomalies, tracheoesophageal fistulas (TEFs), renal anomalies, and limb defects (most often of the radius) are commonly associated and known collectively by the acronym VACTERL. The authors studied these nonrandomly associated birth defects to determine if a further relationship exists between VACTERL association and the presence of a tethered spinal cord (TSC). METHODS: From 2001 to 2004, 12 patients with VACTERL association who were treated operatively by a single pediatric surgeon underwent magnetic resonance (MR) imaging to evaluate the intraspinal abnormalities that may cause tethering. Three patients were excluded from the study due to complications after surgery for TEF. Coincidentally, these three patients did not have imperforate ani. In the remaining nine patients, seven had associated urogenital anomalies, and six of these also had high-type imperforate ani. Five of the six patients and the one patient with low-type imperforate anus and a urogenital anomaly were found to have TSCs. In the remaining two patients without urogenital anomalies there was a high-type imperforate anus without a TSC in one patient and a low-type imperforate anus with a TSC in the other. All seven patients with TSCs underwent successful untethering. The lesions contributing to TSC included terminal filum lipomas (TFLs) in five patients, an intramedullary ependymal cyst in one patient, and a lipo-meningomyelocele in another patient. CONCLUSIONS: The authors found that in patients with VACTERL association there was a high incidence of TSC (seven of nine patients) if an imperforate anus was present as one of the anomalies. In patients with VACTERL association and urogenital anomalies, the incidence of TSC was even higher (86%). Five of the seven cases of TSC in the present study were caused by a TFL, a lesion that can be easily and safely managed surgically. The authors conclude that MR imaging is essential for ruling out the possibility of a TSC in patients with VACTERL association combined with urogenital anomalies or an imperforate anus. PMID- 17465386 TI - Outcome following surgery for temporal lobe epilepsy with hippocampal involvement in preadolescent children: emphasis on mesial temporal sclerosis. AB - OBJECT: The authors conducted a multiinstitutional, retrospective analysis to better define outcome and prognostic indicators for temporal lobe epilepsy surgery for suspected mesial temporal sclerosis (MTS) in young children. METHODS: Data were collected for all children undergoing temporal resections at four epilepsy centers over approximately 10 years. Children with a histopathological diagnosis of neoplasm were excluded. Forty-nine patients (28 boys and 21 girls) were included in the study. Their mean age at surgery was 9.1 years (range 1.25 13.9 years). The mean age at seizure onset was 3.2 years (range birth-10 years). Histopathological examination demonstrated MTS in 26 cases, gliosis in nine, dysplasia in five, gliosis with dysplasia in four, and nonspecific or normal findings in five. Forty-one anterior temporal lobectomies (nine tailored) and eight selective amygdalohippocam-pectomies were performed (28 left side, 21 right side). Twenty-nine children (59.2%) underwent invasive monitoring. Operative complications included extraaxial hematomas (two cases), cerebrospinal fluid leaks (two cases), and hydrocephalus (one case), each in children undergoing invasive monitoring. The mean duration of follow up was 26.4 months (range 5-74 months) overall and 23.9 months (range 6-74 months) for the Engel Class I subgroup. Outcomes at the most recent follow-up examination were categorized as Engel Class I-II in 31 (63.3%) of 49 children overall, 20 (76.9%) of 26 children with confirmed MTS, four (36.4%) of 11 children with gliosis, and four (57.1%) of seven children with dysplasia. All patients who underwent selective amygdalohippocampectomies had confirmed MTS and Engel Class I outcomes. Patients with more than one seizure type (p = 0.048) or moderate to severe developmental delay (p = 0.03) had significantly worse outcomes (Engel Class III or IV). Age at seizure onset, age at surgery, and duration of seizure disorder were not significantly related to outcome. There was a trend for bilateral or extratemporal findings on electroencephalography (EEG) (p = 0.157), high preoperative seizure frequency (p = 0.097), and magnetic resonance (MR) imaging findings inconsistent with MTS (p = 0.142) to be associated with worse outcome, although it did not reach statistical significance. In only 12 (46.1%) of the 26 patients with confirmed MTS was the condition prospectively diagnosed on preoperative MR imaging. CONCLUSIONS: Younger children with temporal lobe epilepsy have satisfying surgical outcomes, particularly when MTS is present. Magnetic resonance imaging may not be as sensitive in detecting MTS in children as in older patients. Negative predictors identified include multiple seizure types and preoperative developmental delay. Multifocal or bilateral EEG findings, high preoperative seizure frequency, and MR imaging findings inconsistent with MTS also independently suggested worse outcome. PMID- 17465387 TI - Fetal spina bifida. PMID- 17465388 TI - Fetal spina bifida in a mouse model: loss of neural function in utero. AB - OBJECT: The devastating neurological deficit associated with myelomeningocele has previously been assumed to be a direct and inevitable consequence of the primary malformation-failure of neural tube closure. An alternative view is that secondary damage to the pathologically exposed spinal cord tissue in utero is responsible for the neurological deficiency. If the latter mechanism were shown to be correct, it would provide an objective rationale for the performance of in utero surgery for myelomeningocele, because coverage of the exposed spinal cord could be expected to alleviate or perhaps prevent neurodegeneration. To examine this question, the authors studied the development of neuronal connections and neurological function of mice during fetal and neonatal stages in a genetic model of exposed lumbosacral spina bifida. METHODS: The persistently exposed spinal cord of mouse fetuses carrying both curly tail and loop-tail mutations exhibited essentially normal anatomical and functional hallmarks of development during early gestation (embryonic Days 13.5-16.5), including sensory and motor projections to and from the cord. A significant proportion of fetuses with spina bifida at early gestation exhibited sensorimotor function identical to that seen in age-matched healthy controls. However, at later gestational stages, increasing neurodegeneration within the spina bifida lesion was detected, which was paralleled by a progressive loss of neurological function. CONCLUSIONS: These findings provide support for the hypothesis that neurological deficit in human myelomeningocele arises following secondary neural tissue destruction and loss of function during pregnancy. PMID- 17465389 TI - Glutaric aciduria type 1 presenting as bilateral subdural hematomas mimicking nonaccidental trauma. Case report and review of the literature. AB - Glutaric aciduria type 1 (GA1) is a rare neurometabolic disorder with characteristic neuroimaging and clinicopathological features. The authors describe a case of GA1 in a 7-month-old girl presenting with macrocephaly and bilateral subdural hematomas (SDHs) who was initially evaluated for nonaccidental trauma (NAT). Bilateral subdural drains were placed because of significant mass effect from the chronic SDHs, with subsequent neurological and neuroimaging documented improvement. Clinical and neuroimaging findings led to further laboratory investigation to confirm the diagnosis of GA1, after which a specialized low-protein diet was initiated. After a thorough investigation, NAT was ruled out. At the follow-up examination, the patient experienced improvement in her symptoms and resolution of the bilateral subdural collections. The presence of bilateral SDHs in an infant raises the suspicion of NAT and presents a difficult diagnostic challenge because of the legal and social implications. Glutaric aciduria type 1 should be considered in the differential diagnosis of bilateral SDHs, and an evaluation should be performed. The authors review the clinical manifestations, diagnosis, medical and surgical management, and specific considerations regarding GA1, including misdiagnosis of NAT. PMID- 17465390 TI - Engorged epidural venous plexus and cervical myelopathy due to cerebrospinal fluid overdrainage: a rare complication of ventricular shunts. Case report. AB - The authors report on a 17-year-old boy with cervical myelopathy from dilated epidural veins due to cerebrospinal fluid (CSF) overdrainage. The patient had a long-standing subdural-peritoneal shunt and presented with incapacitating spastic tetraparesis. Magnetic resonance imaging revealed significant cervical spinal cord compression from a markedly dilated epidural venous plexus. The shunt was externalized so that CSF flow dynamics could be assessed, and the patient was found to have low intracranial pressure (ICP). The patient was gradually acclimated to higher ICPs, and a new shunt was placed with an antisiphon device and a programmable valve set at the higher pressure. Postoperatively the child experienced significant clinical improvement, and reduction of spinal cord compression was evident on images. Compensatory engorgement of the epidural venous plexus due to long-term shunt usage should be considered in the differential diagnosis when cervical myelopathy due to a dilated epidural venous plexus is present. PMID- 17465391 TI - Giant intracranial teratoma with epignathus in a neonate. Case report and review of the literature. AB - Epignathus, a congenital oropharyngeal teratoma, is a rare clinical entity with variable clinical outcomes described in the literature. Even fewer cases of epignathus with intracranial extension have been reported, all with poor outcomes. In this manuscript, the authors present a case of epignathus with intracranial extension, emphasizing clinical presentation, imaging findings, a staged surgical approach, multidisciplinary management, and outcome. PMID- 17465392 TI - Modified encephaloduroarteriosynangiosis with bifrontal encephalogaleoperiosteal synangiosis for the treatment of pediatric moyamoya disease. Technical note. AB - The authors describe a modified technique of encephaloduroarteriosynangiosis (EDAS) with bifrontal encephalogaleoperiosteal synangiosis (EGPS) and present the preliminary results of the procedure. Between January 2004 and June 2005 the authors performed modified EDAS with bifrontal EGPS in 17 patients with moyamoya disease. Surgical results were evaluated in terms of clinical outcomes, changes visible on neuroimages, extent of revascularization noted on angiograms, and hemodynamic changes demonstrated on single-photon emission computed tomography (SPECT) scans. The follow-up period ranged from 6 to 21 months (mean 11.5 months). The overall clinical outcomes were excellent or good in 15 patients (88.2%) and poor in two (11.8%). The overall morbidity rate was 5.9% (one of 17 patients). Based on changes in the anterior cerebral artery (ACA) and middle cerebral artery (MCA) territories after surgery, as shown on SPECT scans following administration of acetazolamide, 14 patients (82.4%) exhibited an improved vascular reserve capacity in both the ACA and MCA territories. It is the authors' opinion that wide covering of the cortex is necessary for sufficient revascularization. In the present study they demonstrate that modified EDAS with bifrontal EGPS is a safe and efficient surgical approach that covers not only the MCA territory but also the ACA territory. PMID- 17465393 TI - Cervical intramedullary tuberculoma in an infant. Case illustration. PMID- 17465394 TI - Subarachnoid air mimicking a basilar apex aneurysm. Case illustration. PMID- 17465395 TI - [Crenotherapy and upper airways diseases. Consensus Conference]. PMID- 17465396 TI - Morphological and chemical studies of pathological human and mice brain at the subcellular level: correlation between light, electron, and nanosims microscopies. AB - Neurodegenerative diseases induce morphological and chemical alterations in well characterized regions of the brain. Understanding their pathological processes requires the use of methods that assess both morphological and chemical alterations in the tissues. In the past, microprobe approaches such as scanning electron microscopy combined with an X-ray spectrometer, Proton induced X-ray emission, secondary ion mass spectrometry (SIMS), and laser microprobe mass analysis have been used for the study of pathological human brain with limited success. At the present, new SIMS instruments have been developed, such as the NanoSIMS-50 ion microprobe, that allow the simultaneous identification of five elements with high sensitivity, at subcellular spatial resolution (about 50-100 nm with the Cs(+) source and about 150-200 nm with O(-) source). Working in scanning mode, 2D distribution of five elements (elemental maps) can be obtained, thus providing their exact colocalization. The analysis can be performed on semithin or ultrathin embedded sections. The possibility of using transmission electron microscopy and SIMS on the same ultrathin sections allows the correlation between structural and analytical observations at subcellular and ultrastructural level to be established. Our observations on pathological brain areas allow us to establish that the NanoSIMS-50 ion microprobe is a highly useful instrument for the imaging of the morphological and chemical alterations that take place in these brain areas. In the human brain our results put forward the subcellular distribution of iron-ferritin-hemosiderin in the hippocampus of Alzheimer disease patients. In the thalamus of transgenic mice, our results have shown the presence of Ca-Fe mineralized amyloid deposits. PMID- 17465397 TI - Alcohols as electrophiles in C--C bond-forming reactions: the hydrogen autotransfer process. AB - The hydrogen autotransfer process involves an initial oxidative hydrogen elimination, followed by different types of reactions, and is completed with a reductive hydrogen addition to give the final product. The sequence allows the alkylation of different nucleophilic agents using environmentally benign alcohols as electrophiles, mild conditions, and soft bases, with water produced as the only waste material. Recent examples of modulating the organometallic catalyst have also lent themselves to expansion of the range of available substrates, as described in this Minireview. PMID- 17465398 TI - ADAM 12 as a second-trimester maternal serum marker in screening for Down syndrome. AB - BACKGROUND: ADAM 12 is a placenta-derived glycoprotein that is involved in growth and differentiation. The maternal serum concentration of ADAM 12 is a potential first-trimester maternal serum marker of Down syndrome (DS). Here we examine the potential of ADAM 12 as a second-trimester maternal serum marker of DS. MATERIALS AND METHODS: The concentration of ADAM 12 was determined in gestational week 14 19 in 88 DS pregnancies and 341 matched control pregnancies. Medians of normal pregnancies were established by polynomial regression and the distribution of log(10) MoM ADAM 12 values in DS pregnancies and controls determined. Correlations with alpha-fetoprotein (AFP) and free beta-human chorionic gonadotrophin (free beta-hCG) were established and used to model the performance of maternal serum screening with ADAM 12 in combination with other second trimester serum markers. RESULTS: The ADAM 12 maternal serum concentration was significantly increased with a median MoM of 1.85 and a mean log(10) MoM (SD) of 0.268 (0.2678) compared to a mean log(10) MoM (SD) of 0.013 (0.4318) in controls. ADAM 12 correlated with maternal weight and ethnicity (with the serum concentration increased in Afro-Caribbeans), but neither with maternal age nor gestational age, and only marginally with AFP (r(DS) = 0.078, r(controls) = 0.093) and free beta-hCG (r(DS) = 0.073, r(controls) = 0.144. The increase in detection rate-for a false positive rate of 5%--by adding ADAM 12 to the double test (AFP + free beta-hCG) was 4%, similar to that of adding uE3 to the double test. CONCLUSION: ADAM 12 is an efficient second-trimester marker for DS. Further studies should be conducted to determine whether it may be a useful additional or alternative marker to those currently used in the second-trimester. PMID- 17465403 TI - Recent progress in the catalytic synthesis of imidazoles. AB - Imidazole is one of the important heteroaromatic compounds that have found a broad range of uses in, for example, pharmaceutical and industrial applications. Several conventional methodologies for the synthesis of imidazole rings are known; however, they are limited in terms of substrate generality and are not always efficient. Recent studies toward the development of catalytic reactions have brought significant improvements to the preparation of imidazoles. This Focus Review highlights the very recent progress in the catalytic synthesis of imidazoles, particularly those involving the formation of five-membered ring cores. PMID- 17465404 TI - Bioinspired nanodevice based on the folic acid/titanium dioxide system. AB - A new bioinspired nanomaterial has been obtained by chemisorption of folic acid onto nanocrystalline titanium dioxide. The organic chromophore is linked with the semiconductor surface via the glutamate chain and anchored with the carboxylate group. The geometry and electronic structure of the chromophore was studied in detail with DFT. Photoelectrochemical studies revealed photosensitization of the new material towards visible light. The photoelectrodes composed of the folic acid/titanium dioxide hybrid material generated photocurrent over a 300-600-nm window. Moreover, the direction of the photocurrent could be changed from anodic to cathodic and vice versa by application of the appropriate photoelectrode potential. Photoelectrochemical and spectroscopic studies allowed the elucidation of the mechanism of photocurrent switching. Photoelectrodes composed of folate modified titanium dioxide may serve as a simple model of optoelectronic switches and may constitute the basis for molecular photoelectronic devices. PMID- 17465405 TI - Folding dynamics of 10-residue beta-hairpin peptide chignolin. AB - Short peptides that fold into beta-hairpins are ideal model systems for investigating the mechanism of protein folding because their folding process shows dynamics typical of proteins. We performed folding, unfolding, and refolding molecular dynamics simulations (total of 2.7 micros) of the 10-residue beta-hairpin peptide chignolin, which is the smallest beta-hairpin structure known to be stable in solution. Our results revealed the folding mechanism of chignolin, which comprises three steps. First, the folding begins with hydrophobic assembly. It brings the main chain together; subsequently, a nascent turn structure is formed. The second step is the conversion of the nascent turn into a tight turn structure along with interconversion of the hydrophobic packing and interstrand hydrogen bonds. Finally, the formation of the hydrogen-bond network and the complete hydrophobic core as well as the arrangement of side chain-side-chain interactions occur at approximately the same time. This three step mechanism appropriately interprets the folding process as involving a combination of previous inconsistent explanations of the folding mechanism of the beta-hairpin, that the first event of the folding is formation of hydrogen bonds and the second is that of the hydrophobic core, or vice versa. PMID- 17465406 TI - Carbonyldinitrosyltris(fluorosulfato)tungstate(II) and -molybdate(II) anions: synthesis, spectroscopy, and density functional theory calculations. AB - Carbonyldinitrosyltris(fluorosulfato)tungstate(II) and -molybdate-(II) anions, [fac-M(CO)(NO)2(SO3F)3]- (M=W, Mo), which are novel weakly coordinating anions that contain a metal carbonyl/nitrosyl moiety, have been generated in fluorosulfonic acid and completely characterized by multinuclear NMR, IR, and Raman spectroscopy as well as ESI mass spectrometry. ESI MS measurements performed for the first time on a superacidic solution system unambiguously reveal the formation of the monoanionic, mononuclear W and Mo complexes formulated as [M(CO)(NO)2(SO3F)3]- (M=W, Mo). Multinuclear NMR spectroscopic studies at natural abundance and 13C and 15N enrichment clearly indicate the presence of one CO ligand, two equivalent NO ligands, and two types of nonequivalent SO3F- groups in a 2:1 ratio. The IR and Raman spectra reveal that the two equivalent NO ligands have a cis conformation, thus indicating a fac structure. Density functional calculations at the B3LYP level of theory predict that these anions have a singlet ground state (1A) with a Cs symmetry along with C-O and N-O vibrational frequencies that are in agreement with the experimental observations. Mulliken population analysis shows that the monovalent negative charge is dispersed on the bulky sphere, the surface of which is covered by all the negatively charged O and F atoms with charge densities much lower than SO3F-, suggesting that [fac-M(CO)(NO)2(SO3F)3]- (M=W, Mo) are weakly nucleophilic and poorly coordinating anions. PMID- 17465407 TI - Phosphorus ligands with a large cavity: synthesis of triethynylphosphines with bulky end caps and application to the rhodium-catalyzed hydrosilylation of ketones. AB - Trialkynylphosphines substituted with bulky triarylsilyl groups at the alkyne termini were synthesized. The new phosphines P(C[triple chemical bond]CSiAr(3))(3) (Ar=3,5-tBu(2)-4-MeOC(6)H(2), 3,5-(Me(3)Si)(2)C(6)H(3)) are uncrowded near the phosphorus atom but bulky in the distal region. As a result, they contain a large cavity, at the bottom of which the phosphine lone-pair electrons are located. The compounds are stable to oxidation by air and hydrolysis. DFT calculations suggested that the triethynylphosphines are good pi acceptor ligands, comparable with P(OAr)(3). The trialkynylphosphines reacted with [{RhCl(cod)}(2)] (P/Rh=1.1:1) to give selectively the monophosphine-rhodium complex [RhCl(cod)P(C[triple chemical bond]CSiAr(3))(3)]. X-ray crystal-structure analysis revealed that the {RhCl(cod)} fragment is fully accommodated by the cavity of the phosphine ligand. Compared to the effect of analogues with smaller end caps and PPh(3), the trialkynylphosphines accelerated markedly the rhodium catalyzed hydrosilylation of ketones with a triorganosilane. It is proposed that the higher catalytic activity observed with the holey phosphines is a result of the preferential formation of a monophosphine-rhodium species. PMID- 17465408 TI - Reactivity differences between carbon nano onions (CNOs) prepared by different methods. AB - The carbon nanoparticles obtained from either arcing of graphite under water or thermal annealing of nanodiamonds are commonly called carbon nano onions (CNOs), or spherical graphite, as they are made of concentric fullerene cages separated by the same distance as the shells of graphite. A more careful analysis reveals some dramatic differences between the particles obtained by these two synthetic methods. Physicochemical methods indicate that the CNOs obtained from nanodiamonds (N-CNOs) are smaller and contain more defects than the CNOs obtained from arcing (A-CNOs). These properties explain the enhanced reactivity of the N CNOs in cycloaddition and oxidation reactions, as well as in reactions involving radicals. Given the easier functionalization of the N-CNOs, they are the most obvious choice for studying the potential applications of these multi-shelled fullerenes. PMID- 17465409 TI - Modular synthesis and dynamics of a variety of donor-acceptor interlocked compounds prepared by click chemistry. AB - A series of donor-acceptor [2]-, [3]-, and [4]rotaxanes and self-complexes ([1]rotaxanes) have been synthesized by a threading-followed-by-stoppering approach, in which the precursor pseudorotaxanes are fixed by using Cu(I) catalyzed Huisgen 1,3-dipolar cycloaddition to attach the required stoppers. This alternative approach to forming rotaxanes of the donor-acceptor type, in which the donor is a 1,5-dioxynaphthalene unit and the acceptor is the tetracationic cyclophane cyclobis(paraquat-p-phenylene), proceeds with enhanced yields relative to the tried and tested synthetic strategies, which involve the clipping of the cyclophane around a preformed dumbbell containing pi-electron-donating recognition sites. The new synthetic approach is amenable to application to highly convergent sequences. To extend the scope of this reaction, we constructed [2]rotaxanes in which one of the phenylene rings of the tetracationic cyclophane is perfluorinated, a feature which significantly weakens its association with pi electron-rich guests. The activation barrier for the shuttling of the cyclophane over a spacer containing two triazole rings was determined to be (15.5+/-0.1) kcal mol(-1) for a degenerate two-station [2]rotaxane, a value similar to that previously measured for analogous degenerate compounds containing aromatic or ethylene glycol spacers. The triazole rings do not seem to perturb the shuttling process significantly; this property bodes well for their future incorporation into bistable molecular switches. PMID- 17465410 TI - Formal total synthesis of (-)-apicularen a by a strategy based on ring-closing metathesis and transannular cyclization. AB - A formal synthesis of (-)-apicularen A, a potent antitumor agent with unique biological properties, has been completed in a 15-step sequence starting from a known, enantiomerically pure hydroxyepoxide, which was generated by using the Jacobsen hydrolytic-kinetic-resolution methodology. The 12-membered macrocyclic lactone in the target was constructed by ring-closing metathesis, and the trans tetrahydropyran ring system was created through the transannular etherification of a hydroxyalkene. PMID- 17465416 TI - Simultaneous chiral separation and determination of ephedrine alkaloids by MEKC ESI-MS using polymeric surfactant I: method development. AB - In this work, simultaneous separation of eight stereoisomers of ephedrine and related compounds ((+/-)-ephedrine, (+/-)-pseudoephedrine, (+/-)-norephedrine and (+/-)-N-methylephedrine) was accomplished using a polymeric chiral surfactant, i.e. polysodium N-undecenoxycarbonyl-L-leucinate (poly-L-SUCL) by chiral (C)MEKC ESI-MS. The conditions of CMEKC were first investigated. The baseline separation of all eight stereoisomers of ephedrine and related compounds was achieved under optimum CMEKC conditions (35 mM poly-L-SUCL, 15 mM NH(4)OAc, pH 6.0, 30% v/v ACN, 30 kV and 20 degrees C) in less than 30 min. Next, a central composite design for response surface modeling has been described to evaluate the electrospray chamber parameters and the sheath liquid conditions. Optimum mass abundance of stereoisomers of ephedrine and related compounds was observed using the spray chamber parameters, namely 250 degrees C drying gas temperature and 8 L/min drying gas flow rate at a nebulizer pressure of 4 psi. Furthermore, the experimental design indicates that the optimum mass abundance of the stereoisomers of ephedrine and related compounds can be obtained using a sheath liquid containing 80:20 v/v methanol-water, 5 mM NH(4)OAc at pH 8.5 delivered at 5 microL/min. Finally, compared to MEKC-UV, the use of poly-L-SUCL in MEKC-MS provided significantly higher sensitivity for stereoisomers of ephedrine and related compounds. PMID- 17465417 TI - Identifying bacterial species using CE-MS and SEQUEST with an empirical scoring function. AB - CE-MS/MS analysis of proteolytic digests of bacterial cell extracts was combined with SEQUEST searching and a new scoring system to identify bacteria species in bacterial mixtures. Searches of MS/MS spectra against protein databases enabled the identification of bacterial species by the matching of the proteins associated with the corresponding species. An empirical scoring function was obtained by evaluating the SEQUEST search results of 38 samples that contained single bacterial species. The scoring by the empirical function helped move up the positive identification results from their original positions in the ranking based on Xcorr values alone. Therefore, the identification of bacteria in the samples that contained bacterial mixtures was improved. Bacterial species in 20 bacterial mixtures, including one real sample, were correctly identified by database searches and the new scoring function. PMID- 17465418 TI - Simultaneous chiral separation of ephedrine alkaloids by MEKC-ESI-MS using polymeric surfactant II: application in dietary supplements. AB - Chiral MEKC-MS method was utilized for separation, identification, and quantitation of ten enantiomers of ephedrine and related compounds. Enantioselective separations of all ephedrine alkaloids were accomplished through a combination of polysodium N-undecenoxycarbonyl-L-leucinate (poly-L-SUCL) with 30% v/v ACN. Interestingly, the more hydrophilic stereoisomers were eluted later than the hydrophobic ones indicating that hydrogen bonding interactions are much stronger than hydrophobic interactions in the presence of ACN in chiral MEKC. The method was validated in terms of linearity, LOD, LOQ, precision and robustness. The method was finally used in the analysis of three standard reference materials (SRMs). Results of (-)-ephedrine ranged from 12.49 to 0.24 mg/g, for (+) pseudoephedrine from 4.04 to 0.019 mg/g, for (-)-norephedrine from 0.36 to 0.0031 mg/g, for (+)-norpseudoephedrine from 0.68 to 0.0052 mg/g, for (-) methylephedrine from 1.18 to 0.0092 mg/g and for (+)-methylpseudoephedrine from 0.086 to 0.00037 mg/g in the SRMs. PMID- 17465421 TI - CEC separation of heterocyclic amines using methacrylate monolithic columns. AB - Two methacrylate-based monolithic columns, one with a negatively charged group (sulfonic group) and another with a new monomer N,N-dimethylamino ethyl acrylate (DMAEA), were prepared and tested for the separation of basic compounds by CEC. This new monolithic stationary phase was prepared by the in situ polymerization of DMAEA with butyl methacrylate and ethylene dimethacrylate, using a ternary porogenic solvent consisting of water, 1-propanol and 1,4-butanediol. The performance of this column was evaluated by means of the analysis of a family of heterocyclic amines. Separation conditions such as pH, amount of organic modifier, ionic strength and elution mode (normal or counterdirectional flow) were studied. At the optimal running electrolyte composition, and using the counterdirectional mode, symmetrical electrochromatographic peaks were obtained, with the number of theoretical plates up to 30,000 and a good resolution between closely related peaks. The 2-acrylamido-2-methyl-1-propane-sulfonic acid column was used for CEC-MS, taking advantage of the compatibility of its elution mode (normal flow) with the MS coupling. PMID- 17465422 TI - Development of a new method for analysis of Sudan dyes by pressurized CEC with amperometric detection. AB - A new analytical method, pressurized CEC (pCEC) with amperometric detection (AD) using 1.5 microm RP nonporous silica packed columns has been developed for the rapid separation and determination of four Sudan dyes in hot chilli. The influence of several experimental parameters on the retention behavior has been investigated. The electrochemical oxidation of Sudans I-IV separated by pCEC can be reliably monitored with a carbon electrode at +0.95 V (vs. Ag/AgCl). Fast and efficient separation of the analytes was achieved within 7 min by pCEC under the optimum conditions with an ACN/water (95:5%) mobile phase containing formic acid (pH 4.3), 5% acetone and 0.002% triethylamine using a separation voltage of 12 kV. The detection limits for four Sudan dyes ranged from 8.0 x 10(-7) to 1.2 x 10(-6) mol/L. To evaluate the feasibility and reliability of this method, the proposed pCEC-AD method was further demonstrated with hot chilli samples spiked with Sudan dyes. PMID- 17465423 TI - Simultaneous determination of cefepime and vancomycin in plasma and cerebrospinal fluid by MEKC with direct sample injection and application for bacterial meningitis. AB - A simple MEKC with UV detection at 214 nm for simultaneous analysis of cefepime and vancomycin in plasma and in cerebrospinal fluid (CSF) by direct injection without any sample pretreatment is described. The separation of cefepime and vancomycin from biological matrices was performed at 25 degrees C using a BGE consisting of a Tris buffer with SDS and methanol as the electrolyte solution. Under optimal MEKC conditions for biological samples, good separations with high efficiency and short analysis time are achieved. Several parameters affecting the separation of the drugs from biological matrices were studied, including methanol, pH, and concentrations of the Tris buffer and SDS. The linear ranges of the method for the determination of cefepime and vancomycin in plasma and in CSF using imidazole or cefazolin as an internal standard, respectively, were all over the range of 1-30 microg/mL; the detection limits of cefepime and vancomycin in biological matrices (injection 10 kV, 15 s) were 0.3 and 0.5 microg/mL, respectively. The applicability of the proposed method for the determination of cefepime and vancomycin in plasma and CSF collected after intravenous administration of the drugs in patients with meningitis was demonstrated. PMID- 17465424 TI - Comparative studies of the interaction between ferulic acid and bovine serum albumin by ACE and surface plasmon resonance. AB - Affinity capillary electrophoresis (ACE) was used to study the interaction between ferulic acid (FA) and BSA. The interaction between FA and BSA was facilitated by injecting FA into a BSA-containing running buffer. Both mobility ratio and mobility shift assays were performed to deduce the binding constant (K(b)). However, the K(b )value obtained with the mobility ratio assay was only approximately 20% of that extracted from the mobility shift assay. The former assay yielded a K(b) value (5.6 +/- 0.4 x 10(4) M(-1)), which compares well with the result obtained with surface plasmon resonance (SPR) (5.1 +/- 0.6 x 10(4) M( 1)). The discrepancy between the mobility ratio and mobility shift assays suggests that the data extrapolation from the mobility ratio should be more reliable for cases when both changes in the EOF and viscosity of the running buffer are important. The work demonstrates that ACE, a solution-based technique, and SPR, a technique addressing interfacial processes, are highly complementary to each other and the comparative studies are confirmatory and allow binding constants to be accurately determined. PMID- 17465425 TI - Mixed-metal (platinum, palladium), mixed-pyrimidine (uracil, cytosine) self assembling metallacalix[n]arenes: dynamic combinatorial chemistry with nucleobases and metal species. AB - Reactions between the mononuclear mixed-nucleobase complex [Pt(en)(UH-N1)(CH2 N3)]+ (1; en: ethylenediamine; UH-N1: uracil monoanion bonded through the N1 atom; CH2-N3: neutral cytosine bonded through the N3 atom) and [Pd(II)(en)] or [Pd(II)(2,2'-bpy)] (2,2'-bpy: 2,2'-bipyridine) lead to libraries of compounds of different stoichiometries and different connectivities. In these compounds, the palladium entity binds to or cross-links either the N3 sites of uracil and/or the N1 sites of cytosine, following deprotonation of these positions to give uracil dianions (U) and cytosine monoanions (CH). Cyclic species, which can be considered as metallacalix[n]arenes, have been detected in several cases, with n being 4 and 8. The complexity of the compounds formed not only results from the possibility of the two different nucleobases in building block 1 engaging in different connectivities with the Pd entities, but also from the potential for the formation of oligomers of different sizes and different conformations; in the case of cyclic tetranuclear Pt(2)Pd(2) species, this can, in principle, lead to the various arrangements (cone, partial cone, 1,2-alternate, 1,3-alternate) known from calix[4]arene chemistry. A further complication arises from the fact that, depending on the mutual orientation of the exocyclic groups of the two nucleobases (O2 and O4 of uracil, O2 and N4 of cytosine), these sites can be engaged in additional chelation of [Pd(II)(en)] and [Pd(II)(2,2'-bpy)]. Thus, penta-, hexa-, and octanuclear complexes, Pt(2)Pd(3), Pt(2)Pd(4), and Pt(2)Pd(6), derived from cyclic Pt(2)Pd(2) tetramers have been isolated and characterized. PMID- 17465426 TI - Design and non-covalent DNA binding of platinum(II) metallacalix[4]arenes. AB - A set of cyclic tetranuclear complexes of the metallacalix[4]arene type with formula [{Pt(en)(L)}(4)](4+) (en=ethylenediamine; 2: LH=5-chloro-2 hydroxypyrimidine (5-Cl-Hpymo); 3: LH=5-bromo-2-hydroxypyrimidine (5-Br-Hpymo); 4: LH=5-iodo-2-hydroxypyrimidine (5-I-Hpymo)) have been obtained from the reaction between cis-protected square-planar [Pt(en)(H(2)O)(2)](2+) metal entities and LH in aqueous media. Additionally, the binding properties of 2, 3, 4 and their congener [{Pt(en)(L)}(4)](4+) (1: LH=2-hydroxypyrimidine (Hpymo)) with calf thymus-DNA (ct-DNA) have been studied by using different techniques including circular and linear dichroism (CD and LD, respectively) and UV-visible absorbance spectroscopies, gel electrophoresis, fluorescence competitive-binding studies and atomic force microscopy (AFM). The results are consistent with significant non-covalent interactions taking place between the polynuclear cyclic species and ct-DNA. Moreover, gel electrophoresis, linear dichroism titrations and AFM images of ct-DNA with metallacalixarenes show ct-DNA coiling at low metallacalixarene concentrations and upon subsequent increments in metallacalixarene concentration ct-DNA can be seen to uncoil with concomitant formation of long and inflexible ct-DNA structures. PMID- 17465427 TI - Calix[4]arene-based bis[2]catenanes: synthesis and chiral resolution. AB - The exclusive formation of hydrogen-bonded dimers between tetraaryl and tetratosylurea calix[4]arenes has been used to prepare a series of ten "bisloop" tetraurea calix[4]arenes 3, in which adjacent phenylurea groups are covalently linked through alpha,omega-dioxyalkane chains. This dimerization with tetratosylurea 2 as template preorganizes the alkenyl residues of tetra(m alkenyloxyphenyl) ureas 1 and enables their selective connection in high yield (up to 95 %) by olefin metathesis followed by hydrogenation. The "bisloop" calixarenes 3 also exclusively form heterodimers with 1. Thus, in a separated metathesis/hydrogenation sequence, a series of 14 cyclic bis[2]catenanes 4, in which two calix[4]arenes are connected through their wide rims by two pairs of interlocked rings (total size 29 to 41 atoms), were prepared in yields of up to 97 %. Optical resolution of these chiral bis[2]catenanes was studied by HPLC on chiral stationary phases. The single-crystal X-ray structure of one example (4(P,10)) confirmed the interlocking rings and revealed that the hydrogen-bonded dimeric capsule of the calix[4]arene can be "completely" opened. PMID- 17465428 TI - Rational synthetic tuning between itinerant antiferromagnetism and ferromagnetism in the complex boride series Sc2FeRu(5-n)RhnB2 (0or= 1.5 mg/dL, odds ratio, 16.0) independently predicted postoperative CDAD. CONCLUSION: CDAD should be considered in the differential diagnosis of patients with postoperative diarrhea after LDLT. PMID- 17465451 TI - Preoperative diagnosis of gastrointestinal stromal tumor by endoscopic ultrasound guided fine needle aspiration. AB - AIM: To evaluate the role of endoscopic ultrasonography-guided fine needle aspiration (EUS-FNA) in the preoperative diagnosis of gastrointestinal stromal tumor (GIST). METHODS: From September 2002 to June 2006, Fifty-three consecutive EUS-FNAs of GI tract subepithelial hypoechoic tumors with continuity to proper muscle layer suspected as GIST by standard EUS were evaluated prospectively. The reference standards for the final diagnosis were surgery (n = 31), or clinical follow-up (n=22). Additionally, immunophenotyping of specimens obtained by EUS FNA and surgical resection specimens were compared. RESULTS: In 2 cases puncture was not performed because of anatomical problems. The collection rate of adequate specimens from the GI tract subepithelial hypoechoic tumor with continuity to proper muscle layer was 82% (42/51). The diagnostic rate for the tumor less than 2 cm, 2 to 4 cm, and 4 cm or more were 71% (15/21), 86% (18/21), and 100% (9/9), respectively. In 29 surgically resected cases, the sensitivity, specificity, positive predictive value, negative predictive value, and diagnostic accuracy of EUS-FNA using immunohistochemical analysis of GIST were 100% (24/24), 80% (4/5), 96% (24/25), 100% (4/4), and 97% (28/29), respectively. No major complications were encountered. CONCLUSION: EUS-FNA with immunohistochemical analysis is a safe and accurate method in the pretherapeutic diagnosis of GIST. It should be taken into consideration in decision making, especially in early diagnosis following minimal invasive surgery for GIST. PMID- 17465452 TI - Gastric digestion of pea ferritin and modulation of its iron bioavailability by ascorbic and phytic acids in caco-2 cells. AB - AIM: To understand the digestive stability and mechanism of release and intestinal uptake of pea ferritin iron in caco-2 cell line model. METHODS: Pea seed ferritin was purified using salt fractionation followed by gel filtration chromatography. The bioavailability of ferritin iron was assessed using coupled in vitro digestion/Caco-2 cell model in the presence or absence of ascorbic acid and phytic acid. Caco-2 cell ferritin formation was used as a surrogate marker of iron uptake. Structural changes of pea ferritin under simulated gastric pH were characterized using electrophoresis, gel filtration and circular dichroism spectroscopy. RESULTS: The caco-2 cell ferritin formation was significantly increased (P<0.001) with FeSO(4) (19.3 +/- 9.8 ng/mg protein) and pea ferritin (13.9 +/- 6.19 ng/mg protein) compared to the blank digest (3.7 +/- 1.8 ng/mg protein). Ascorbic acid enhanced while phytic acid decreased the pea ferritin iron bioavailability. However, either in the presence or absence of ascorbic acid, the ferritin content of caco-2 cells was significantly less with pea ferritin than with FeSO(4). At gastric pH, no band corresponding to ferritin was observed in the presence of pepsin either on native PAGE or SDS-PAGE. Gel filtration chromatography and circular dichroism spectroscopy revealed a pH dependent loss of quaternary and secondary structure. CONCLUSION: Under gastric conditions, the iron core of pea ferritin is released into the digestive medium due to acid induced structural alterations and dissociation of protein. The released iron interacts with dietary factors leading to modulation of pea ferritin iron bioavailability, resembling the typical characteristics of non-heme iron. PMID- 17465453 TI - Crohn's disease, fatigue, and infliximab: is there a role for cytokines in the pathogenesis of fatigue? AB - AIM: To study the effect of infliximab on fatigue in relation to cytokine levels in Crohn's disease (CD) patients. METHODS: Fourteen CD patients were blinded for treatment and received placebo at baseline, and infliximab 2 wk later, with a follow-up of 4 wk. Blood samples were drawn on a regular basis, and questionnaires on fatigue, depression, quality of life, and clinical disease activity were completed at regular intervals. RESULTS: After placebo infusion, fatigue scores decreased within 3 d (3.5 points +/- 1.1, P T (34.5%) in ethnic Chinese compared to literature reports on other Asian populations and Caucasians. The frequency of the 516TT genotype is higher in the Han majority (23.1%) than in three other ethnic minority groups (i.e., 7.4%, 9.1% and 15.8%) in southern China. CONCLUSION: This was the first study to document the spectrum of CYP2B6 allelic variants and genotypes in a southern Chinese population. The 516G > T allele is associated with a defective metabolism of efavirenz (EFV), which therefore may predispose to drug toxicity. Treatment regimens for human immunodeficiency virus (HIV) and heroin addiction may need to be optimized in different populations because of the marked variability of the key metabolizing enzyme. PMID- 17465456 TI - Relationship between hepatitis B virus DNA levels and liver histology in patients with chronic hepatitis B. AB - AIM: To study the relationship between hepatitis B virus (HBV) DNA levels and liver histology in patients with chronic hepatitis B (CHB) and to determine the prevalence and characteristics of hepatitis B e antigen (HBeAg) negative patients. METHODS: A total of 213 patients with CHB were studied, and serum HBV DNA levels were measured by the COBAS Amplicor HBV Monitor test. All patients were divided into two groups according to the HBeAg status. The correlation between serum HBV DNA levels and liver damage (liver histology and biochemistry) was explored. RESULTS: Of the 213 patients with serum HBV DNA levels higher than 10(5) copies/mL, 178 (83.6%) were HBeAg positive, 35 (16.4%) were HBeAg negative. The serum HBV DNA levels were not correlated to the age, history of CHB, histological grade and stage of liver disease in either HBeAg negative or HBeAg positive patients. There was no correlation between serum levels of HBV DNA and alanine aminotransferanse (ALT), aspartate aminotransferase (AST) in HBeAg positive patients. In HBeAg negative patients, there was no correlation between serum levels of HBV DNA and AST, while serum DNA levels correlated with ALT (r=0.351, P=0.042). The grade (G) of liver disease correlated with ALT and AST (P<0.05, r=0.205, 0.327 respectively) in HBeAg positive patients. In HBeAg negative patients, correlations were shown between ALT, AST and the G (P<0.01, and r=0.862, 0.802 respectively). HBeAg negative patients were older (35 +/- 9 years vs 30 +/- 9 years, P<0.05 ) and had a longer history of HBV infection (8 +/ 4 years vs 6 +/- 4 years, P<0.05) and a lower HBV DNA level than HBeAg positive patients (8.4 +/- 1.7 Log HBV DNA vs 9.8 +/- 1.3 Log HBV DNA, P<0.001). There were no significant differences in sex ratio, ALT and AST levels and liver histology between the two groups. CONCLUSION: Serum HBV DNA level is not correlated to histological grade or stage of liver disease in CHB patients with HBV DNA more than 10(5) copies/mL. Compared to HBeAg positive patients, HBeAg negative patients are older and have a lower HBV DNA level and a longer HBV infection history. There is no significant difference in sex ratio, ALT and AST levels and liver histology between the two groups. PMID- 17465457 TI - Relationship between preoperative staging by endoscopic ultrasonography and MMP-9 expression in gastric carcinoma. AB - AIM: To investigate the relationship between the staging by endoscopic ultrasonography (EUS) and the expression of carcinoma metastasis associated gene in the patients with gastric carcinoma. METHODS: Sixty-three patients with gastric cancer were diagnosed by electric gastroscopy and EUS. The preoperative staging of gastric cancer was measured by EUS and compared with pathologic staging and MMP-9 expression. Peripheral serum level of MMP-9 was measured with enzyme-linked immunosorbent assay (ELISA), while the expression of MMP-9 protein was tested with immunohistochemistry and hybridization in situ in the gastric carcinoma tissues. RESULTS: The total accuracy of EUS in estimating invasive depth of gastric cancer was 80.95%, while that in estimating lymphatic metastasis was 73.02%. Serum MMP-9 levels were consistent with the expression of MMP-9 protein and MMP-9 mRNA in tissue, a result closely correlated with invasive degree, staging with EUS and lymphatic metastasis in gastric cancer (P<0.05). The total accuracy of estimating invasive depth in gastric cancer was 95.22% using both EUS and MMP-9. CONCLUSION: The MMP-9 level of preoperative serum presents the reference value for preoperative staging by EUS in the patients with gastric cancer. When serum MMP-9 level in gastric cancer is significantly high, physicians should pay closer attention to the metastasis which reaches the serosa or beyond. Combining EUS and MMP-9 improves the accuracy in deciding the invasion and metastasis in the patients with gastric carcinoma. PMID- 17465458 TI - Trans-arterial gene therapy for hepatocellular carcinoma in a rabbit model. AB - AIM: To study the effect of adenovirus (Ad)-p53 gene therapy on hepatocellular carcinoma (HCC) in a rabbit model. METHODS: VX2 tumor was grown in the liver of 24 rabbits. Animals were divided into four groups: group A receiving trans arterial gene therapy (Ad-p53) only, group B receiving combined Ad-p53 therapy and trans-arterial embolization (lipiodol), group C receiving trans-arterial chemoembolization (lipiodol + mitomycin C), control group (D) receiving sodium chloride. Tumor volume (V1) was measured by using MRI (d 13). Interventional procedure was applied (d 14). Tumor volume (V2) was assessed by MRI (d 21) and the mean ratio (V2/V1) was calculated. After the second MRI, specimens of the liver were abstained and examined immunohistochemically using mutant-type p53 antibody. The positive expression was scored. RESULTS: Compared with control group (chi= 3.14 +/- 0.64), therapeutic groups all showed a significant decrease in the tumor growth ratio (P<0.05). A slight difference was found between group A (chi = 2.35 +/- 0.59) and group B (chi = 1.75 +/- 0.28) (P=0.048). No statistically significant difference was observed between group B and group C (chi = 2.00 +/- 0.44). The positive expression rate of mutant-type p53 was the lowest in group B and significantly different between group A and group C (P<0.05). Compared to the control subjects, groups A and C both showed a decrease in the expression of mutant-type p53, but there was no significant difference between them. CONCLUSION: Trans-arterial Ad-p53 gene therapy can reduce tumor growth of HCC in rabbit model. PMID- 17465459 TI - Differences in expression of retinal proteins between diabetic and normal rats. AB - AIM: To compare and identify the differences in expression of retinal proteins between normal and diabetic rats, and to analyze the molecular pathogenetic mechanisms of retinal diseases caused by diabetes. METHODS: Changes in protein expression of retinal tissues from diabetic and normal rats were observed using 2 dimensional polyacrylamide gel electrophoresis (2-DE). Some protein spots exhibiting statistically significant variations (P<0.05) were selected randomly and identified by tandem mass spectrometry and analyzed by bioinformatics. RESULTS: 2-DE showed that the expression was up-regulated in 5 retinal proteins, down-regulated in 23 retinal proteins, and disappeared in 8 retinal proteins. Eight spots were identified from the 36 spots by tandem mass spectrometry (MS/MS) and analyzed by bioinformatics. Guanylate kinase 1, triosephosphate isomerase 1, ATP synthase subunit d, albumin and dimethylarginine dimethylaminohydrolase 2 played an important role in signal transduction. Triosephosphate isomerase 1, crystallin alpha B, ATP synthase subunit d and peroxiredoxin 6 were involved in energy metabolism of retinal tissues. Guanylate kinase 1 played an important role in photoexcitation of retinal rod photoreceptor cells. Whether crystallin beta A1 plays a role in diabetic retinas is unknown so far. CONCLUSION: There are differences in expression of retinal proteins between diabetic and normal rats. These proteins may be involved in the mechanisms and prognosis of retinal diseases caused by diabetes. PMID- 17465460 TI - Gut perforation after orthotopic liver transplantation in adults. AB - AIM: To describe cases of gut perforation after orthotopic liver transplantation. METHODS: Data were collected from our center database and medical records. Six of 187 patients (3.2%) who underwent orthotopic liver transplantation from January to December 2005 developed gut perforation. All patients were male with an average age of 46 years. Modified piggyback liver transplantation was performed at the Organ Transplantation Center, First Affiliated Hospital, Sun Yat-Sen University. RESULTS: Previous operation, steroid therapy, and prolonged portal venous cross clamp time, poor nutritional status and iatrogenic injury were found to be its ecological factors. The patients with gut perforation were found to have fever, increased leukocytes, mild abdominal pain and tenderness. The median portal venous clamp time was 63 min (range 45-72 min), median cold ischaemia time was 11.3 h (range 7-15 h). Median intraoperative blood loss was 500 mL (range 100 1200 mL) and median operation time was 8.8 h (range 6-12 h). None of the six patients developed acute cellular rejection. White cell count was above 18 x 10(9)/L in five patients (neutrophilic leukocytes were above 90%) and 1.5 x 10(9)/L in one patient. Bacterial culture in drainage liquid revealed enterococci in five patients. Of the 6 patients undergoing orthotopic liver transplantation, 3 survived and 3 died after modified piggyback liver transplantation. CONCLUSION: Gut perforation occurs after orthotopic liver transplantation in adults. A careful and minimal dissection during OLT, longer retention of the stomach tube, and reducing the portal clamp time and steroid dose should be taken into consideration. If gut perforation is not prevented, then early diagnosis, preferably through detection of enterococci may ensure better survival. PMID- 17465461 TI - Presacral ganglioneuroma: a case report and review of literature. AB - Presacral ganglioneuromas are so rare benign tumors that only 17 cases have been reported in the literature. They are abdominal masses growing slowly and differential diagnoses have to be considered. Surgical resection is important for definitive diagnosis because it represents the only therapeutic choice. Because of the benign nature of ganglioneuroma, adjuvant chemo- or radiotherapy is not indicated but regular follow-up is necessary for an early diagnosis of potential local recurrence. We report a case of a 64-year-old man with a presacral ganglioneuroma. PMID- 17465462 TI - A case of oesophageal ulcer developed after taking homeopathic pill in a young woman. AB - Oesophageal ulcers occur mainly as a result of gastro-oesophageal reflux disease (GERD). However, pill-induced oesophageal ulcers are a fairly common event. The lesion is mainly due to entrapment of the pill and/or its chemical composition thereof. This case report describes an oesophageal mucosa ulcer occurred in a healthy 35-year old woman who had no previous history of oesophageal disorders and received homeopathic medication. The present case reveals that pill entrapment can occur even in the oesophagus of healthy young individuals and that oesophageal mucosal ulcer can be triggered by substances generally thought devoid of any potentially mucosal aggressive effect. PMID- 17465463 TI - A case of capecitabine-induced coronary microspasm in a patient with rectal cancer. AB - 5-Fluorouracil (5-FU) is the most frequently used chemotherapy agent concomitant with radiotherapy in the management of patients with rectal cancer. Capecitabine is an oral fluoropyrimidine that mimics the pharmaconkinetics of infusional 5-FU. This new drug is replacing 5-FU as a part of the combined-modality treatment of a number of gastrointestinal cancers. While cardiac events associated with the use of 5-FU are a well known side effect, capecitabine-induced cardiotoxicity has been only rarely reported. Here, we reviewed the case of a patient with rectal cancer who had a capecitabine-induced coronary vasospasm. The most prominent mutation of the dihydropyrimidine dehydrogenase gene was also analyzed. PMID- 17465464 TI - Celiac disease and skin: psoriasis association. PMID- 17465465 TI - Microsatellite instability and MLH1 promoter hypermethylation in colorectal cancer. AB - Colorectal cancer (CRC) is caused by a series of genetic or epigenetic changes, and in the last decade there has been an increased awareness that there are multiple forms of colorectal cancer that develop through different pathways. Microsatellite instability is involved in the genesis of about 15% of sporadic colorectal cancers and most of hereditary nonpolyposis cancers. Tumors with a high frequency of microsatellite instability tend to be diploid, to possess a mucinous histology, and to have a surrounding lymphoid reaction. They are more prevalent in the proximal colon and have a fast pass from polyp to cancer. Nevertheless, they are associated with longer survival than stage-matched tumors with microsatellite stability. Resistance of colorectal cancers with a high frequency of microsatellite instability to 5-fluorouracil-based chemotherapy is well established. Silencing the MLH1 gene expression by its promoter methylation stops the formation of MLH1 protein, and prevents the normal activation of the DNA repair gene. This is an important cause for genomic instability and cell proliferation to the point of colorectal cancer formation. Better knowledge of this process will have a huge impact on colorectal cancer management, prevention, treatment and prognosis. PMID- 17465467 TI - Synergistic effect of oxymatrine and angiogenesis inhibitor NM-3 on modulating apoptosis in human gastric cancer cells. AB - AIM: To investigate the synergistic effect of oxymatrine (OM) and angiogenesis inhibitor NM-3 on modulating apoptosis in human gastric cancer cell lines SGC 7901, MKN-45, MKN-74. METHODS: Human gastric cancer lines SGC-7901, MKN-45, MKN 74 were treated with OM in the absence and presence of NM-3. The inhibitory rates were detected by MTT assay. Synergistic effect of OM and NM-3 on the growth of survivin, bcl-2, bax and p53 in SGC-7901 cells were examined by semiquantitative RT-PCR and Western blotting, and their growth inhibitory effects were also observed on SGC-7901 tumor xenograft in nude mice. RESULTS: OM combined with NM-3 exhibited a synergistic inhibitory effect on the growth of SGC-7901, MKN-45 and MKN-74 cells in a time-dependent manner. Twenty-four hours after treatment with OM, NM-3 alone and their combination, mRNA expression of survivin and bcl-2 in SGC-7901 cells decreased, p53 mRNA expression increased. OM (4 g/L) combined with NM-3 significantly increased the expression of p53 mRNA and decreased the expression of survivin and bcl-2 compared with either agent alone (193% +/- 34% vs 129% +/- 12%; 44% +/- 18% vs 92% +/- 18%; 36 +/- 17% vs 93% +/- 23%, P < 0.05). Western blotting showed that the synergistic effect of OM and NM-3 on protein translation of survivin, bcl-2 and p53 was in accordance with their mRNAs. Furthermore, OM/NM-3 combination obviously exhibited antitumor growth effect in xenografted human gastric cancer cells SGC-7901 compared with either agent alone. CONCLUSION: OM combined with NM-3 has synergistic inhibitory effects on human gastric cancer cells in vitro and can suppress the growth of xenografted human gastric cancer cells SGC-7901 in vivo. PMID- 17465466 TI - A comparative review of HLA associations with hepatitis B and C viral infections across global populations. AB - Hepatitis B (HBV) and hepatitis C (HCV) viral infection or co-infection leads to risk of development of chronic infection, cirrhosis and hepatocellular carcinoma (HCC). Immigration and globalization have added to the challenges of public health concerns regarding chronic HBV and HCV infections worldwide. The aim of this study is to review existing global literature across ethnic populations on HBV and HCV related human leukocyte antigen (HLA) associations in relation to susceptibility, viral persistence and treatment. Extensive literature search was conducted to explore the HLA associations in HBV and HCV infections reported across global populations over the past decade to understand the knowledge status, weaknesses and strengths of this information in different ethnic populations. HLA DR13 is consistently associated with HBV clearance globally. HLADRB1*11/*12 alleles and DQB1*0301 are associated with HBV persistence but with HCV clearance worldwide. Consistent association of DRB1*03 and *07 is observed with HCV susceptibility and non-responsiveness to HBV vaccination across the population. HLA DR13 is protective for vertical HBV and HCV transmission in Chinese and Italian neonates, but different alleles are associated with their susceptibility in these populations. HLA class I molecule interactions with Killer cell immunoglobulin like receptors (KIR) of natural killer (NK) cells modulate HCV infection outcome via regulating immune regulatory cells and molecules. HLA associations with HBV vaccination, interferon therapy in HBV and HCV, and with extra hepatic manifestations of viral hepatitis are also discussed. Systematic studies in compliance with global regulatory standards are required to identify the HLA specific viral epitope, stage specific T cell populations interacting with different HLA alleles during disease progression and viral clearance of chronic HBV or HCV infections among different ethnic populations. These studies would facilitate stage specific therapeutic strategies for clearance of HBV and HCV infections or co-infections across global populations and aid in identification of HBV-HCV combined vaccine. HLA associations of chronic HBV or HCV development with confounding host factors including alcohol, drug abuse, insulin resistance, age and gender are lacking and warrant detailed investigation across global populations. PMID- 17465468 TI - Association of serum vascular endothelial growth factor-C and lymphatic vessel density with lymph node metastasis and prognosis of patients with gastric cancer. AB - AIM: To investigate whether serum vascular endothelial growth factor-C (SVEGF-C), VEGF-C, and lymphatic vessel density (LVD) in tumor tissues are related to lymph node metastasis (LNM) and prognosis in gastric cancer. METHODS: SVEGF-C levels of 80 gastric cancer patients and 20 healthy donors were examined using ELISA. VEGF C expression and LVD were examined using immunohistochemical staining. Kaplan Meier survival analysis was performed to determine their influence on the prognosis of the patients. RESULTS: The SVEGF-C level in gastric cancer patients (595.9 +/- 201.0 ng/L) was significantly higher (P = 0.000) than controls (360.0 +/- 97.4 ng/L). Both SVEGF-C and LVD were significantly higher in poorly differentiated adenocarcinomas, T3 and T4, LNM, distant metastasis, and pTNM groups III and IV (P = 0.000). The sensitivity and specificity of SVEGF-C for predicting LNM were 82.8% and 81.8%, respectively (cut-off = 542.5 ng/L). The positive expression rate of VEGF-C was significantly higher in cancerous than in normal tissues (65% vs 20%; P = 0.001). VEGF-C expression up-regulation was significantly related to differentiation, depth of invasion, LNM, distant metastasis, and pTNM stage (P = 0.000). LVD was 10.7 +/- 3.1/200 HP in the experimental group vs 4.9 +/- 1.3/200 HP in controls (P = 0.000); LVD in cancerous tissues with and without LNM was 12.0 +/- 2.7/200 HP vs 7.6 +/- 0.5/200 HP, respectively (P = 0.000). SVEGF-C and LVD were significantly higher in VEGF-C positive than in negative patients (P = 0.000); SVEGF-C level was related to LVD (P = 0.000). Kaplan-Meier survival analysis factors predicating poor prognosis were: SVEGF-C level (P = 0.001), VEGF-C expression and LVD (both P = 0.000). CONCLUSION: SVEGF-C level, VEGF-C and LVD are related to LNM and poor prognosis of patients with gastric cancer. SVEGF-C may be a biomarker for LNM in gastric cancer. PMID- 17465469 TI - Cloning and expression of special F protein from human liver. AB - AIM: To clone human liver special F protein and to express it in a prokaryotic system. METHODS: Total RNA was isolated from human liver tissue and first-strand cDNA was reverse transcribed using the PCR reverse primer. Following this, cDNA of the F protein was ligated into the clone vector pUCm-T. The segment of F protein's cDNA was subcloned into the expression vector pET-15b and transformed into E. coli BL21 (DE3) pLyss. Isopropy-beta-D-thiogalactoside (IPTG) was then used to induce expression of the target protein. RESULTS: The cDNA clone of human liver special F protein (1134bp) was successfully produced, with the cDNA sequence being published in Gene-bank: DQ188836. We confirmed the expression of F protein by Western blot with a molecular weight of 43 kDa. The expressed protein accounted for 40% of the total protein extracted. CONCLUSION: F protein expresses cDNA clone in a prokaryotic system, which offers a relatively simple way of producing sufficient quantities of F protein and contributes to understanding the principal biological functions of this protein. PMID- 17465470 TI - Sonographic fatty liver and hepatitis B virus carrier status: synergistic effect on liver damage in Taiwanese adults. AB - AIM: To examine the epidemiology of hepatitis B virus carrier status (HBVC) and sonographic fatty liver (SFL) in Taiwanese adults, and to evaluate their possible interaction in inducing liver damage (LD). >From an epidemiological viewpoint, we analyzed previous studies which indicated that fatty liver sensitizes host immune response to HBV infection and enhances liver damage. METHODS: A cross-sectional retrospective analysis of health records including medical history, physical examination, abdominal sonogram, blood biochemistry and hepatic virological tests. We utilized the Student's t-test, chi-square, multivariate logistic regression and synergy index to assess risks for LD. RESULTS: Among a total of 5406 Taiwanese adults (mean age 46.2 years, 51.5% males), the prevalence of LD, HBVC and SFL were 12.3%, 15.1% and 33.4%, respectively; 5.1% of participants had SFL plus HBVC. Multivariate logistic regression analysis demonstrated that male gender (odds ratio (OR) = 2.8, 95% confidence interval (CI): 2.3-3.5), overweight state (OR = 1.6, 95% CI: 1.3-2.0), HBVC (OR = 2.5, 95% CI: 2.0-3.1) and SFL (OR = 4.2, 95% CI: 2.2-5.3) were independently associated with LD. Synergism analysis showed that the adjusted OR for LD in adults with HBVC-alone was 3.3 (95% CI: 2.4 4.6), SFL-alone, 4.7 (95% CI: 3.7-6.1) and combined HBVC and SFL, 9.5 (95% CI: 6.8-13.3); the synergy index was 1.4 (95% CI: 1.001-2.0). CONCLUSION: In Taiwanese adults, SFL plus HBVC have a significant synergistic association with LD. PMID- 17465471 TI - Clinicopathological study of solid and pseudopapillary tumor of pancreas: emphasis on magnetic resonance imaging findings. AB - AIM: To report the clinicopathological features and magnetic resonance imaging (MRI) findings of solid and pseudopapillary tumor (SPT) of pancreas. METHODS: From 1981 to 2005, 26 surgically treated cases of SPT were retrospectively reviewed. MRI findings of the latest 11 consecutive SPT cases were investigated. RESULTS: There were 25 women and one man having SPT (median age: 23 year) with a median tumor size of 7.5 cm. Among them, nine patients developed solid pseudopapillary carcinoma. During the median follow-up period of 66 mo, the 5 year survival rate of the 26 SPT patients was 96.2%. Three MRI features were proposed including Type 1 image, displaying SPT with completely solid part. All SPT patients with type 1 image were detected incidentally. Type 2 image displays of SPT with solid mass hemorrhage and type 3 image with massive hemorrhage. All the eight SPT patients with type 2 and 3 images suffered abdominal pain due to hemorrhage from SPT. CONCLUSION: SPT had a favorable survival rate irrespective of surgical procedures, malignancy, and MRI findings, however, MRI could reliably correlate with its clinicopathological features. PMID- 17465472 TI - Appropriateness of colonoscopy: diagnostic yield and safety in guidelines. AB - AIM: To evaluate if the guidelines for the appropriateness of performing colonoscopy by American Society for Gastrointestinal Endoscopy (AGSE) and Italian Society of Digestive Endoscopy (SIED) yield a good diagnostic efficacy and do not present risks of missing important colonic pathologies in an Italian population sample. METHODS: A total of 1017 consecutive patients (560 men and 457 women; mean age 64.4 +/- 16 years) referred to an open-access endoscopy unit for colonoscopy from July 2004 to May 2006 were evaluated according to ASGE and SIED guidelines for appropriateness of performing the procedure. Diagnostic yield was defined as the percentage of relevant colonic pathologies of the total number of colonoscopies performed. RESULTS: About 85.2% patients underwent colonoscopy that was considered appropriate based on at least one ASGE or SIED criterion, while it was considered inappropriate for 14.8% of patients. The diagnostic yield of colonoscopy was significantly higher for appropriate colonoscopies (26.94% vs 10.6%, P < 0.001) than for inappropriate colonoscopies (5.3%). There was no missed colorectal cancer following the ASGE/SIED criteria. CONCLUSION: ASGE/SIED guidelines have shown a good diagnostic yield and the rate of missing relevant colonic pathologies seems very low. Unfortunately, the percentage of inappropriate referrals for colonoscopy in an open-access endoscopy system is still high, despite the number of papers published on the issue and the definition of international guidelines. Further steps are required to update and standardize the guidelines to increase their diffusion and to promote educational programs for general practitioners. PMID- 17465473 TI - Role of intravenously administered hyoscine butyl bromide in retrograde terminal ileoscopy: a randomized, double-blinded, placebo-controlled trial. AB - AIM: To evaluated the role of hyoscine butyl bromide in facilitating retrograde ileoscopy. METHODS: Retrograde terminal ileoscopy was attempted in 200 consecutive patients undergoing colonoscopy. After intubation of the cecum and visualization of the ileocecal valve, butyl bromide injection or normal saline was given intravenously to the patients in a double blind random fashion. The pulse rate and oxygen saturation were measured continuously. After completion of the procedure, endoscopists were then asked to score the ease of intubation and the ease of visualization of the terminal ileum on a visual scale of 1 to 10. The patients were also asked to score the pain after receiving hyoscine butyl bromide injection on a score of 1 to 10. RESULTS: Terminal ileoscopy could be performed in 188 patients. The mean (SD) visual analogue score for the ease of intubation of the cecum was 7.4 (0.65) in the injection group and 5.9 (0.8) in the placebo group (P < 0.001). The mean (SD) length of ileum visualized in the injection group was 14.4 (3.3) cm and 10.4 (2.7) cm in the placebo group (P < 0.001). The mean (SD) visual analogue score for ease of visualization of the terminal ileum was 7.5 (0.69) in the injection group and 5.9 (0.7) in the placebo group (P < 0.001). The pain score experienced by the patients was 6.5 (0.7) in the injection group and 6.7 (0.69) in the placebo group (P < 0.008). Although the pulse rate increased significantly in patients receiving the drug, no statistically significant difference was noted in the oxygen saturation between the two groups either before or after administration of the drug. No complications were observed in either of the groups. CONCLUSION: Hyoscine butyl bromide injection is a useful adjunct in helping the intubation and visualization of terminal ileum during colonoscopy. PMID- 17465474 TI - Clinical heterogeneity in autoimmune acute liver failure. AB - AIM: To describe the outcome and prognosis in a cohort of patients with acute liver failure due to autoimmune hepatitis without liver transplantation. METHODS: A retrospective trial was conducted in 11 patients with acute liver failure due to autoimmune hepatitis who attended the Instituto Nacional de Ciencias Medicas y Nutricion Salvador Zubiran. Demographic, biochemical and severity indexes, and treatment and outcome were assessed. RESULTS: Among the 11 patients, with a median age of 31 years, 72% had inflammatory response syndrome, and six patients received corticosteroids. The mortality rate within four weeks was 56%, and the one-year survival was 27%. In the survivors, severity indexes were lower and 83% received corticosteroids. CONCLUSION: We observed a relatively high survival rate in patients with acute liver failure due to autoimmune hepatitis. This survival rate could be influenced by severity of the disease and/or use of corticosteroids. PMID- 17465475 TI - Prevalence and risk factors of asymptomatic hepatitis C virus infection in Egyptian children. AB - AIM: To identify the prevalence, risk factors and manifestations of asymptomatic hepatitis C virus (HCV) infection in Egyptian children. METHODS: Children at the age of 1-9 years were screened for HCV antibodies and alanine aminotransferase (ALT) levels. Every child with elevated ALT and/or detectable HCV antibodies was tested for HCV RNA by RT-PCR and compared with two negative controls for risk factors and signs and symptoms of liver disease. RESULTS: We screened 1042 children, six of them had elevated ALT, negative HCV antibody and positive RNA, likely representing acute hepatitis C cases. Fifteen children were HCV seropositive, 5 of them were HCV RNA positive. Asymptomatic HCV infection was present in 2.02% (positive results for either HCV antibodies or HCV-RNA or both). Symptoms such as diarrhea, abdominal pain, history of fatigue and school absence because of illness and risk factors such as dental care were significantly more common among HCV positive cases than among controls. None of the HCV positive children was diagnosed as having signs of advanced liver disease upon clinical or ultrasonographic examination. CONCLUSION: Asymptomatic HCV infection is detectable in 2.02% Egyptian children. PMID- 17465476 TI - Cyclic vomiting syndrome in children: experience with 181 cases from southern Iran. AB - AIM: To evaluate the clinical presentation, response to prophylactic therapy and outcome of children with cyclic vomiting syndrome (CVS) in Shiraz, Iran. METHODS: During a period of 11 years (March 1994 to March 2005), 181 consecutive children with a final diagnosis of CVS were evaluated, treated and followed in our center. Patients were randomized to receive either amitriptyline or propranolol as prophylactic treatments. RESULTS: There were 88 boys and 93 girls with mean age of onset of symptoms of 4.9 +/- 3.3 years (range, neonatal period to 14 years), the mean age at final diagnosis was 6.9 years (range, 1.5 to 14), and the mean duration between the onset of the first attack and the final diagnosis of CVS was 2 +/- 1.81 years (range, 1/6 to 8). The mean duration of each attack was 4.26 days (range, from few hours to 10 d) and the mean interval between the attacks was 1.8 mo (range, 1 wk to 12 mo). The time of onset of the attacks was midnight to early morning in about 70% of cases. Amitriptyline was effective in 46 out of 81 (56%) patients (P < 0.001). Propranolol appeared to have a superior action and was effective in 74 out of 83 (92%) patients (P < 0.0001). CONCLUSION: There is a significant lag time between the onset of clinical symptoms and the final diagnosis of CVS in our area. In patients with typical clinical presentations of CVS, who are examined by an experienced physician, invasive workup is not necessary. Propranolol appears more effective than amitriptyline for prophylactic use in children with CVS. PMID- 17465477 TI - Knowledge, attitude and practice related to liver fluke infection in northeast Thailand. AB - AIM: To investigate the knowledge, attitude and practice (KAP) in prevention and control of liver fluke infection in northeast Thailand. METHODS: A descriptive KAP survey pertaining to liver fluke infection was carried out in June 2005 to October 2006 using structured questionnaires. Data were collected by questionnaires consisting of general parameters, knowledge, attitude, practice, and a history of participation in the prevention and control of liver fluke infection. RESULTS: A total of 1077 persons who were inter-viewed and completed the questionnaires were enrolled in the study. The majority were females (69.5%) and many of them were 15-20 years of age (37.26%). The questionnaires revealed that information resources on liver fluke infection included local public health volunteers (31.37%), public health officers (18.72%), televisions (14.38%), local heads of sub-districts (12.31%), doctors and nurses (9.18%), newspaper (5.72), internets (5.37%), and others (12.95%). Fifty-five point eleven percent of the population had a good level of liver fluke knowledge concerning the mode of disease transmission and 79.72% of the population had a good level of prevention and control knowledge with regards to defecation and consumption. The attitude and practice in liver fluke prevention and control were also at a good level with a positive awareness, participation, and satisfaction of 72.1% and 60.83% of the persons studied. However, good health behavior was found in 39.26% and 41.42% of the persons studied who had unhygienic defecation and ate raw cyprinoid's fish. The result also showed that 41.25% of the persons studied previously joined prevention and control campaigns. CONCLUSION: The persons studied have a high level of liver fluke knowledge and positive attitude. However, improvement is required regarding personal hygiene specifically with hygienic defecation and consumption of undercooked fish. PMID- 17465478 TI - Immunoscintigraphy of local recurrent rectal cancer with 99mTc-labeled anti-CEA monoclonal antibody CL58. AB - AIM: To explore a specific diagnostic method for local recurrent rectal cancer. METHODS: Immunoscintigraphy with (99m)Tc-labeled anti-CEA monoclonal antibody (MoAb) CL-58 was performed for patients suspected of having a postoperative local recurrent rectal cancer and the findings were compared with the results of conventional imaging and pathology. RESULTS: A total of 36 patients with a suspected local recurrent rectal cancer underwent immunoscintigraphy with (99m)Tc conjugated CL58. Local recurrence of rectal cancer was identified in 31 patients and established in 30 during operation, endoscopy and pathological examination. No local recurrence was found in 5 patients without specific accumulation of (99m)Tc during the follow-up. Immunoscintigraphy had a positive rate of 86.11%, a specificity of 83.33%, and a sensitivity of 100%. CONCLUSION: Immunoscintigraphy has a highly specific and predictive value for detecting local recurrent rectal cancer, especially after abdominal perineal resection (APR). PMID- 17465479 TI - Screening for metronidazole-resistance associated gene fragments of H pylori by suppression subtractive hybridization. AB - AIM: To screen for metronidazole (MTZ)-resistance associated gene fragments of H pylori by suppression subtractive hybridization (SSH). METHODS: Five MTZ resistant (tester, T) and 1 MTZ-susceptible (driver, D) clinical H pylori isolates were selected. Genomic DNAs were prepared and submitted to Rsa I digestion. Then two different adaptors were ligated respectively to the 5'-end of two aliquots of the tester DNA fragments and SSH was made between the tester and driver DNAs. The specific inserts of tester strains were screened and MTZ resistance related gene fragments were identified by dot blotting. RESULTS: Among the randomly selected 120 subtractive colonies, 37 DNA fragments had a different number of DNA copies (> or = 2 times) in resistant and susceptible strains and 17 of them had a significantly different number of DNA copies (> or = 3 times). Among the sequences obtained from the 17 DNA fragments, new sequences were found in 10 DNA fragments and duplicated sequences in 7 DNA fragments, representing respectively the sequences of depeptide ABC transporter periplasmic dipeptide binding protein (dppA), permease protein (dppB), ribosomal protein S4 (rps4), ribonuclease III (rnc), protease (pqqE), diaminopimelate epimerase (dapF), acetatekinase (ackA), H pylori plasmid pHP51 and H pylori gene 1334. CONCLUSION: Gene fragments specific to MTZ-resistant H pylori strains can be screened by SSH and may be associated with MTZ-resistant H pylori. PMID- 17465480 TI - Contribution of mononuclear bone marrow cells to carbon tetrachloride-induced liver fibrosis in rats. AB - AIM: To study the inhibitory effect of mononuclear bone marrow cell (BMC) transplantation on carbon tetrachloride (CCl(4)) -induced liver fibrosis in rats. METHODS: Rat liver fibrosis models were induced by CCl(4) and alcohol administration. After 8 wk, twenty rats were randomly allocated into treatment group (n = 10) and control group (n = 10). BMC were infused into the rats in treatment group via the portal vein, while heparinized saline was infused in control group. CCl(4) was hypodermically injected into the rats twice a week for 4 wk. At the end of wk 12, all rats were humanely sacrificed. Liver samples were taken and stained with HE or Masson trichrome. The general conditions, liver fibrosis (hydroxyproline and collagen fibre) and liver pathological grades in rats were evaluated. RESULTS: The general conditions of the rats in treatment group improved markedly, but not in control group. Hydroxyproline was 504.6 +/- 128.8 microg/g in treatment group, and 596.0 +/- 341.8 microg/g in control group. The percentage of collagen fibre was 3.75% +/- 0.98% in treatment group and 5.02% +/- 0.44% in control group. There was a significant difference between the two groups (P < 0.05). Liver pathological grade decreased from grade IV to grade III partially in treatment group (P < 0.05) with no obvious improvement in control group (P > 0.05). There was a significant difference between treatment group and control group (P < 0.05). CONCLUSION: Transplantation of BMC can improve liver fibrosis due to chronic liver injury in rats. PMID- 17465481 TI - A clinicopathological analysis in unsuspected gallbladder carcinoma: a report of 23 cases. AB - AIM: To study the clinicopathological characteristics of unsuspected gallbladder carcinoma (UGC). METHODS: We retrospectively studied 23 cases of UGC in Tongji Hospital, and compared their clinicopathological characteristics with 33 cases of preoperatively diagnosed gallbladder carcinoma (PDGC). RESULTS: The proportion of UGC coexisting with cholecystolithiasis was significantly higher than that of PDGC (chi(2) = 13.53, P < 0.01). The infection rate of hepatitis B virus was 21.74% (5/23) in UGC and 30.30% (10/33) in PDGC. Nine (39.13%) of 23 patients with UGC and 8/33 (24.24) PDGC had contact with schistosome pestilent water. The rate of multiple pregnancies was 56.52% (13/23) in the patients with UGC and 42.42% (14/33) in PDGC. The primary location of the UGC was mostly in the neck and body of the gallbladder, and that of the PDGC was often in the body and bottom. The incidence of Nevin stage I and II UGC was significantly higher than that of PDGC (chi(2) = 4.44, P < 0.05 and chi(2) = 4.96, P < 0.05) while that of Nevin stage V UGC was significantly lower than that of PDGC (chi(2) = 7.59, P < 0.01). According to the grading of carcinoma, the incidence of well differentiated UGC was significantly higher than that of PDGC (chi(2) = 4.16, P < 0.05), and that of poorly-differentiated UGC was significantly lower than that of PDGC (chi(2) = 4.48, P < 0.05). CONCLUSION: There are different characteristics between UGC and PDGC, such as in primary location, malignant degree and incidence of coexistence with cholecystolithiasis. Cholecystolithiasis, hepatitis B, schistosome and multiple pregnancies were high risk factors for gallbladder carcinoma. PMID- 17465482 TI - Significance of blood analysis in hemophiliacs co-infected with human immunodeficiency virus and hepatitis viruses. AB - AIM: To study the effect of hepatitis virus infection on cirrhosis and liver function markers in HIV-infected hemophiliacs. METHODS: We have analyzed the immunological, liver function and cirrhosis markers in a cohort of hemophiliacs co-infected with human immunodeficiency virus (HIV) and hepatitis viruses. RESULTS: There was no difference in immunological markers among co-infected patients and patients infected with HIV only and those co-infected with one or more hepatitis virus. Although liver function and cirrhosis markers remained within a normal range, there was a worsening trend in all patients co-infected with hepatitis virus C (HCV), which was further exacerbated in the presence of additional infection with hepatitis virus B (HBV). CONCLUSION: Co-infection with HIV, HBV and HCV leads to worsening of hyaluronic acid and liver function markers. Increases in serum hyaluronic acid may be suggestive of a predisposition to liver diseases. PMID- 17465483 TI - Vagina vasorum dissection during D2 lymphadenectomy for gastric carcinoma. AB - AIM: To explore the relationship between metastasis and vagina vasorum in the progress of gastric carcinoma and to find some facts and references for gastric surgeons. METHODS: One hundred and seven specimens of left or right gastric arteries (55 left and 52 right) were gathered from 59 patients undergoing radical gastrectomy for gastric carcinoma. All the frozen specimens were cut into 3 microm-thick sections and stained with hematoxylin-eosin (HE) and immunohistochemical method separately. Cytokeratin (CK) and mesothelial cells (MC) were stained with immunohistochemical method. Cancer cells inside vagina vasorum were detected and the structure of artery wall was observed under microscope. RESULTS: Metastatic cancer cells or tubercles were found inside vagina vasorum in some stage III or IV specimens, but not in stage I or II specimens. Tumor cells in vagina vasorum were CK positive in 26 specimens of 14 tumors. Among them, stage III was found in 4 specimens of 2 tumors, and stage IV in 22 specimens of 12 tumors. None of these specimens was positive for MC. The positive rate of CK increased with TNM staging. Compared with the lower part, tumors in the upper and middle parts of stomach were more likely to metastasize into vagina vasorum. CONCLUSION: Vagina vasorum dissection should be performed during D2 lymphadenectomy for TNM stage III or IV gastric carcinoma. PMID- 17465484 TI - Significance of hepatitis B virus surface antigen, hepatitis C virus expression in hepatocellular carcinoma and pericarcinomatous tissues. AB - AIM: To investigate the correlation between hepatitis B virus surface antigen (HBsAg), hepatitis C virus (HCV) expression in hepatocellular carcinoma (HCC), the HAI score of the noncancerous region of the liver and the serum Alpha fetoprotein (AFP) level. METHODS: The patterns of HBsAg and HCV in 100 cases of HCC and their surrounding liver tissues were studied on paraffin-embedded sections with immuno-histochemistry, the histological status was determined by one pathologist and one surgeon simultaneously using the hepatitis activity index (HAI) score, and AFP was detected by radioimmunity. The study included 100 consecutive patients who underwent curative resection for HCC. Based on HBsAg and HCV expression, the patients were classified into 4 groups: patients positive for HBsAg (HBsAg group), patients positive for HCV (HCV group), patients negative for both HCV and HBsAg (NBNC group) and patients positive for both HBsAg and HCV (BC group). RESULTS: The BC group had significantly higher HAI scores than the other three groups. (BC > HCV > HBsAg > NBNC). HBV and HCV virus infection was positively correlated with HAI (r(s) = 0.39, P = 0.0001). The positive rate of AFP (85.7%) and the value of AFP (541.2 ng/mL) in the group with HBV and HCV co infection were the highest among the four groups. The positive rate (53.3%) of AFP and the value of AFP ( 53.3 ng/mL) in the group with none-infection of HBV and HCV were the lowest. HBV and HCV virus infection was positively correlated with AFP(r(s) = 0.38, P = 0.0001). CONCLUSION: The AFP increase in patients with liver cancer was positively correlated with the infection of HBV and HCV. The serum AFP elevation by the infection of HBV and HCV is one of mechanisms which lead to hepatocarcinogenesis, and the antivirus intervening treatment of hepatitis is significant for the prognosis of liver cancer. From our Spearman's rank correlation analysis, we can conclude that the severity of virally induced inflammation is correlated with HBsAg and HCV expression in HCC tissues and noncancerous tissues. Prior co-infection of HBV in HCV patients may be an adverse risk factor for intrahepatic inflammation. PMID- 17465485 TI - Diphyllobothrium latum: a case of an incidental finding. AB - Colorectal cancer screening can be performed by fecal occult blood testing, sigmoidoscopy, double contrast barium enema or colonoscopy. Colonoscopy has the significant advantage that polyps can be removed during the same procedure. Occasionally, colonoscopy can also reveal unexpected findings, including parasitic infections, even in asymptomatic patients. Tapeworms or cestodes are hermaphroditic parasites, which can live for considerable periods of time in the human gastrointestinal tract. Fish tapeworm (Diphyllobothrium species) are endemic in various parts of the world, the commonest species being Diphyllobothrium latum. Humans are the main definitive host for D. Latum and the majority of individuals harbouring the parasite are asymptomatic, while 40% of infected individuals may have low vitamin B12 levels. We describe a case of D. latum infection found on routine colonoscopic screening for colorectal cancer in an asymptomatic patient, which was successfully treated with praziquantel. The infection likely arose following raw fish (sushi) consumption. PMID- 17465486 TI - Localized amyloidosis of the stomach: a case report. AB - We report an unusual case of primary amyloidosis of the stomach in a patient complaining of dyspeptic symptoms. The diagnosis was confirmed histologically and other gastrointestinal site or systemic involvement was ruled out. Uncharacteristic dyspeptic symptoms may hide this rare metabolic disease. PMID- 17465487 TI - Unusual causes of intrahepatic cholestatic liver disease. AB - We report five cases with unusual causes of intrahepatic cholestasis, including consumption of Teucrium polium (family Lamiaceae) in the form of tea, Stauffer's syndrome, treatment with tamoxifen citrate for breast cancer, infection with Coxiella Burnetii (acute Q fever), and infection with Brucella melitensis (acute brucellosis). PMID- 17465488 TI - Relationship between the pathogenesis of sarcoidosis and hepatitis C. PMID- 17465489 TI - Short mucin 1 alleles are associated with low virulent H pylori strains infection. PMID- 17465490 TI - A different approach for sterilization of liver hydatid cysts. PMID- 17465491 TI - Recent advances in basic and clinical aspects of inflammatory bowel disease: which steps in the mucosal inflammation should we block for the treatment of inflammatory bowel disease? AB - There are four steps in the interaction between intestinal microbes and mucosal inflammation in genetically predisposed individuals from the viewpoints of basic and clinical aspects of inflammatory bowel disease (IBD). The first step is an interaction between intestinal microbes or their components and intestinal epithelial cells via receptors, the second step an interaction between macrophages and dendritic cells and mucosal lymphocytes, the third step an interaction between lymphocytes and vascular endothelial cells, and the fourth step an interaction between lymphocytes and granulocytes producing proinflammatory cytokines or free radicals and mucosal damage and repair. Recent therapeutic approaches for IBD aim to block these four steps in the intestinal inflammation of patients with IBD. PMID- 17465492 TI - Effect of smoking on inflammatory bowel disease: Is it disease or organ specific? AB - Smoking is an important environmental factor in inflammatory bowel disease (IBD) with differing effects in ulcerative colitis (UC) and Crohn's disease (CD). Never smoking and formerly smoking increase the risk of UC, whereas smoking exacerbates the course of CD. The potential mechanisms involved in this dual relationship are yet unknown. A reasonable assumption is that smoking has different effects on the small and large intestine. This assumption is based on animal and human studies that show that the effects of smoking/nicotine on CD and UC depend on the site of inflammation and not on the type of disease. PMID- 17465493 TI - Celiac disease in the developing countries: a new and challenging public health problem. AB - In the past, celiac disease was believed to be a chronic enteropathy, almost exclusively affecting people of European origin. The availability of new, simple, very sensitive and specific serological tests (anti-gliadin, anti-endomysium and anti-transglutaminase antibody assays) have shown that celiac disease is common not only in Europe and in people of European ancestry but also in the developing countries where the major staple diet is wheat (Southern Asia, the Middle East, North West and East Africa, South America), both in the general population and in the groups at risk. Gluten intolerance thus appears to be a widespread public health problem and an increased level of awareness and clinical suspicion are needed in the New World where physicians must learn to recognize the variable clinical presentations (classical, atypical and silent forms) of celiac disease. In the developing countries, both serological screening in the general population and serological testing in groups at risk are necessary for an early identification of celiac patients. The gluten-free diet poses a challenging public health problem in the developing countries, especially since commercial gluten-free products are not available. PMID- 17465494 TI - In vivo subsurface morphological and functional cellular and subcellular imaging of the gastrointestinal tract with confocal mini-microscopy. AB - AIM: To evaluate a newly developed hand-held confocal probe for in vivo microscopic imaging of the complete gastrointestinal tract in rodents. METHODS: A novel rigid confocal probe (diameter 7 mm) was designed with optical features similar to the flexible endomicroscopy system for use in humans using a 488 nm single line laser for fluorophore excitation. Light emission was detected at 505 to 750 nm. The field of view was 475 microm multiply 475 microm. Optical slice thickness was 7 microm with a lateral resolution of 0.7 microm. Subsurface serial images at different depths (surface to 250 microm) were generated in real time at 1024 multiply 1024 pixels (0.8 frames/s) by placing the probe onto the tissue in gentle, stable contact. Tissue specimens were sampled for histopathological correlation. RESULTS: The esophagus, stomach, small and large intestine and meso, liver, pancreas and gall bladder were visualised in vivo at high resolution in n = 48 mice. Real time microscopic imaging with the confocal mini-microscopy probe was easy to achieve. The different staining protocols (fluorescein, acriflavine, FITC-labelled dextran and L. esculentum lectin) each highlighted specific aspects of the tissue, and in vivo imaging correlated excellently with conventional histology. In vivo blood flow monitoring added a functional quality to morphologic imaging. CONCLUSION: Confocal microscopy is feasible in vivo allowing the visualisation of the complete GI tract at high resolution even of subsurface tissue structures. The new confocal probe design evaluated in this study is compatible with laparoscopy and significantly expands the field of possible applications to intra-abdominal organs. It allows immediate testing of new in vivo staining and application options and therefore permits rapid transfer from animal studies to clinical use in patients. PMID- 17465495 TI - Therapeutic and prophylactic thalidomide in TNBS-induced colitis: synergistic effects on TNF-alpha, IL-12 and VEGF production. AB - AIM: To evaluated the therapeutic and prophylactic effect of thalidomide on 2, 4, 6-trinitrobenzene sulfonic acid (TNBS)-induced colitis. Thalidomide has been reported to downregulate the expression of tumor necrosis factor alpha (TNF alpha), IL-12, and vascular endothelial growth factor (VEGF), hallmarks of intestinal inflammation in Crohnos disease (CD). METHODS: Male Wistar rats were divided in five groups of ten animals each. Four groups received a rectal infusion of TNBS in ethanol. The first group was sacrificed 7 d after colitis induction. The second and third groups received either thalidomide or placebo by gavage and were sacrificed at 14 d. The fourth group received thalidomide 6 h before TNBS administration, and was sacrificed 7 d after induction. The fifth group acted as the control group and colitis was not induced. Histological inflammatory scores of the colon were performed and lamina propria CD4+ T cells, macrophages, and VEGF+ cells were detected by immunohistochemistry. TNF-alpha and IL-12 were quantified in the supernatant of organ cultures by ELISA. RESULTS: Significant reduction in the inflammatory score and in the percentage of VEGF+ cells was observed in the group treated with thalidomide compared with animals not treated with thalidomide. Both TNF-alpha and IL-12 levels were significantly reduced among TNBS induced colitis animals treated with thalidomide compared with animals that did not receive thalidomide. TNF-alpha levels were also significantly reduced among the animals receiving thalidomide prophylaxis compared with untreated animals with TNBS-induced colitis. Intestinal levels of TNF-alpha and IL-12 were significantly correlated with the inflammatory score and the number of VEGF+ cells. CONCLUSION: Thalidomide significantly attenuates TNBS induced colitis by inhibiting the intestinal production of TNF-alpha, IL-12, and VEGF. This effect may support the use of thalidomide as an alternate approach in selected patients with CD. PMID- 17465496 TI - Effect of pre-moxibustion on apoptosis and proliferation of gastric mucosa cells. AB - AIM: To observe the effects of pre-moxibustion on apoptosis and proliferation of gastric mucosal cell in rats with stress-induced ulcer, and to analyze the relationship between those effects and the expression of heat-shock protein 70 (HSP70). METHODS: Sixty healthy Sprague Dawley rats were randomly assigned into four groups, namely group A, B, C and D. The animal model of stress ulcer was established by water immersion and restraint stress. The rats in group A, B, and D served as the restraint, model, and non-acupoint controls, respectively, while those in group C received moxibustion at Zusanli and Liangmen points. Immunohistochemical methodology was used to detect the expression of HSP70, apoptosis index (AI, multiply 10(-6)/microm(2)) and proliferation index (PCNA-LI, multiply 10(-6)/microm(2)). The mucosal expression of transforming growth factor alpha(TGF-alpha) was detected by radioimmunoassay. RESULTS: Moxibustion at Zusanli and Liangmen points significantly decreased the gastric injury and the apoptosis of gastric mucosal cells, while markedly increased the mucosal expression of TGF-alpha and HSP70 as well as the proliferation of gastric mucosal cells. Compared with group A, ulcer index (UI) (26.8 +/- 9.8 vs 12.0 +/- 5.9, P < 0.01), AI (9.6 +/- 4.2 vs 4.4 +/- 2.6, P < 0.05) and expression of HSP70 (9.6 +/- 4.2 vs 4.4 +/- 2.6, P < 0.05) were significantly increased, but the content of TGF-alpha (104.7 +/- 51.2 pg/mL vs 254.0 +/- 86.9 pg/mL, P < 0.01) and PCNA-LI (6.9 +/- 4.7 vs 14.9 +/- 4.6, P < 0.05) were significantly decreased in group B. However, ulcer index values (UI) and AI were obviously lower in group C compared to groups B and D (14.1 +/- 5.4 vs 26.8 +/- 9.8 and 26.2 +/- 7.7, P < 0.01; 3.0 +/- 1.6 vs 9.6 +/- 4.2 and 8.2 +/- 5.2, P < 0.05, respectively), but content of TGF-alpha (237.0 +/- 72.6 pg/mL vs 104.7 +/- 51.2 pg/mL and 154.1 +/- 61.3 pg/mL, P < 0.01) and expression of HSP70 (0.13 +/- 0.03 vs 0.08 +/- 0.06 and 0.06 +/- 0.04, P < 0.05) were higher in group C. Furthermore, the PCNA-LI was significantly higher in group C than in group B (21.6 +/- 4.1 vs 6.9 +/- 4.7, P < 0.01). CONCLUSION: Moxibustion at Zusanli and Liangmen points has a protective effect on rats gastric mucosa in stress-induced gastric ulcer, which is closely related to its actions in promoting synthesis of TGF-alpha and proliferation of gastric mucosal cells, suppressing gastric mucosal cell apoptosis, and up regulating HSP70 expression. PMID- 17465497 TI - Small sphincterotomy combined with papillary dilation with large balloon permits retrieval of large stones without mechanical lithotripsy. AB - AIM: To introduce a new method: small endoscopic sphincterotomy (ES) combined with endoscopic papillary large balloon dilation (SES + EPLBD) to treat patients with large biliary stones. METHODS: Retrieval of large biliary stones was performed in 88 patients. Mean stone size was 14 +/- 3 mm and mean number of stones was 2.5 +/- 3.5. Firstly, ES with a small incision was performed. Next, endoscopic papillary dilation was performed with a large balloon to slowly match the size of the bile duct. Stones were then retrieved from the biliary duct with a balloon and a basket. RESULTS: Stone retrieval was successful in all cases except one cystic duct stone case without the need to crush large stones. Mean procedure time was 30 +/- 5 min. Dilating the papillary orifice with a large balloon made it possible to remove large stones smoothly without crushing them. After dilation with the large balloon, there were some instances of oozing, but no perforations. One instance of post-procedural pancreatitis (1%) occurred. CONCLUSION: SES + EPLBD was effective for the retrieval of large biliary stones without the use of mechanical lithotripsy. PMID- 17465498 TI - Safety and success of precut biliary sphincterotomy: Is it linked to experience or expertise? AB - AIM: To determine the rates of success and complications of precut biliary sphincterotomy (PBS) based on prior experience and to compare the complication rates between PBS and standard endoscopic sphincterotomy (ES). METHODS: A retrospective evaluation of prospectively collected non-randomized data at an academic tertiary referral center. The study included all patients in an eight year period who underwent PBS and ES by a single endoscopist who had no formal training in PBS. The main outcome measures of the study were success and complications of PBS with a comparison to complications of ES. RESULTS: A total of 2939 endoscopic retrograde cholangiopancreatographies (ERCPs) were performed during the study period, including 818 (28%) ES and 150 (5%) PBS procedures. Selective biliary cannulation via PBS was successful at the first attempt in 75% of the patients. Cannulation was achieved in an additional 13% of the patients at a subsequent attempt (total 87%). Complication rate from PBS was 45% higher than ES, but did not differ significantly [7% (10/50) vs 5% (38/818), P = 0.29]. None of the complications from PBS was severe. A significant trend towards increasing success existed with regard to the endoscopistos first attempt at precut (P = 0.0393, Cochran-Armitage exact test for trend, Z = -1.7588). CONCLUSION: Despite the lack of specific training in this technique, PBS was performed with a high success rate and a complication rate similar to or less than reports from other experienced centers. These results suggest that endoscopic experience and perhaps innate endoscopic skill may play an important role in the outcome of this procedure. PMID- 17465499 TI - CCL2-2518 A/G and CCR2 190 A/G do not influence the outcome of hepatitis C virus infection in the Spanish population. AB - AIM: To assess whether CCL2 or interactions between this chemokine and its receptor (CCR2) are associated with outcomes of chronic hepatitis C and with responses to antiviral therapy. METHODS: Two hundred and eighty-four patients with chronic hepatitis C and 193 non-infected matched controls were included in this study. Patients were categorized according to their Scheuer score of hepatic fibrosis as F0-F2 (n = 202) or F3-F4 (n = 82) and according to their response to anti-Hepatitis C virus (HCV) therapy as sustained response (SR, n = 101) or non sustained response (NSR, n = 98). Genotyping of the -2518 (A/G) CCL2 was performed using PCR-RFLP, genotyping of the 190 (A/G) CCR2 using a PCR-ARMS system, and genotyping of the rs3138042 (G/A) CCR2 using Taqman probes. RESULTS: Univariate analyses identified 4 parameters (infection duration time, viral genotype, gender and AST levels) that tended to influence fibrosis and 7 parameters (CCL2G, CCL2ACCR2A, viremia levels, fibrosis stage, viral genotype, infection duration time and AST levels) that significantly influenced or tended to influence response to treatment. Multivariate analysis identified gender and AST levels as parameters that independently influenced fibrosis stage and viral genotype and infection duration time were the two parameters that independently influenced response to treatment. CONCLUSION: Our results indicate that the mutations studied in the gene pair CCL2/CCR2 do not play a major role in the outcome and response to treatment for HCV infection in the Spanish population. PMID- 17465500 TI - Evaluation of 5 versus 10 granulocyteaphaeresis sessions in steroid-dependent ulcerative colitis: a pilot, prospective, multicenter, randomized study. AB - AIM: To evaluate the efficacy of 5 compared to 10 granulocyteaphaeresis sessions in patients with active steroid-dependent ulcerative colitis. METHODS: In this pilot, prospective, multicenter randomized trial, 20 patients with moderately active steroid-dependent ulcerative colitis were randomized to 5 or 10 granulocyteaphaeresis sessions. The primary objective was clinical remission at wk 17. Secondary measures included endoscopic remission and steroid consumption. RESULTS: Nine patients were randomized to 5 granulocyteaphaeresis sessions (group 1) and 11 patients to 10 granulocyteaphaeresis sessions (group 2). At wk 17, 37.5% of patients in group 1 and 45.45% of patients in group 2 were in clinical remission. Clinical remission was accompanied by endoscopic remission in all cases. Eighty-six percent of patients achieving remission were steroid-free at wk 17. Daily steroid requirements were significantly lower in group 2. Eighty-nine per cent of patients remained in remission during a one year follow-up. One serious adverse event, not related to the study therapy, was reported. CONCLUSION: Granulocyteaphaeresis is safe and effective for the treatment of steroid-dependent ulcerative colitis. In this population, increasing the number of aphaeresis sessions is not associated with higher remission rates, but affords a significant steroid-sparing effect. PMID- 17465501 TI - In vitro study of lovastatin interactions with amiodarone and with carbon tetrachloride in isolated rat hepatocytes. AB - AIM: To investigate the interactions at a metabolic level between lovastatin, amiodarone and carbon tetrachloride in isolated rat hepatocytes. METHODS: For cell isolation two-step collagenase liver perfusion was performed. Lovastatin was administered alone in increasing concentrations (1 mumol/L, 3 mumol/L, 5 mumol/L and 10 mumol/L) and in combination with CCl(4) (86 mumol/L). The cells were also pretreated with 14 mumol/L amiodarone and then the other two compounds were added. RESULTS: Lovastatin promoted concentration-dependent significant toxicity estimated by decrease in cell viability and GSH level by 45% and 84%, respectively. LDH-activity increased by 114% and TBARS content by 90%. CCl(4)induced the expected severe damage on the examined parameters. CCl(4) induced toxicity was attenuated after lovastatin pretreatment, which was expressed in less increased values of LDH activity and TBARS levels, as well as in less decreased cell viability and GSH concentrations. However, the pretreatment of hepatocytes with amiodarone abolished the protective effect of lovastatin. CONCLUSION: We suggest that the observed cytoprotective effect was due to interactions between lovastatin, CCl(4) and amiodarone at a metabolic level. PMID- 17465502 TI - No association of the cytotoxic T-lymphocyte associated gene CTLA4 +49A/G polymorphisms with Crohn's disease and ulcerative colitis in Hungarian population samples. AB - AIM: The goal of the current work was to analyse the prevalence of the +49A/G variant of the cytotoxic T-lymphocyte antigen 4 gene (CTLA4) in Hungarian patients with Crohnos disease (CD) and ulcerative colitis (UC). METHODS: A total of 130 unrelated subjects with CD and 150 with UC, and 170 matched controls were genotyped for the single nucleotide polymorphism (SNP). The genotypes were determined by using PCR/RFLP test. RESULTS: The G allele frequency and the prevalence of the GG genotype were 38.1% and 12.3% in the CD group, 40.6% and 18.6% in the UC patients, and 37.4% and 15.9% in the control group, respectively. CONCLUSION: The results of the current study show that carriage of the +49G SNP in heterozygous or in homozygous form does not confer risk either for CD or for UC in the Hungarian population. PMID- 17465503 TI - Expression of p21(WAF1) is related to acetylation of histone H3 in total chromatin in human colorectal cancer. AB - AIM: To explore the relationship between acetylation of histone in total chromatin and p21(WAF1) expression regulation in human colorectal carcinoma. METHODS: We analyzed the expression of tumor suppressor gene p21(WAF1) mRNA by RT PCR or real-time PCR in 33 samples of colorectal cancerous tissue, corresponding para-cancerous tissue and normal colorectal mucosa, and also examined the level of acetylated histone H3 in total chromatin using Western blotting. RESULTS: The expression level of p21(WAF1) mRNA was significantly lower in colorectal cancerous tissue from 33 patients than in para-cancerous tissue and normal colorectal mucosa (2377.95 +/- 865.80 vs 3216.58 +/- 1149.42 and 3541.61 +/- 1433.17 respectively, P < 0.01). In addition, when p21(WAF1) mRNA expression was undectectable or at very low level (50% less than that in adjacent tissue and normal colorectal mucosa) in all tissues, the level of acetylated histone H3 in colorectal cancerous tissue was significantly lower than that in corresponding para-cancerous tissue and normal colorectal mucosa in five of seven (71.43%) cases. The transcriptional level of p21(WAF1) in colorectal carcinoma might not be associated with its biological behaviors. CONCLUSION: The down-regulation of p21(WAF1) transcription is involved in the tumorigenesis and development of colorectal carcinoma. The down-expression of p21(WAF1) mRNA in colorectal carcinoma might be associated with histone hypoacetylation in chromatin but not with biological behaviors. PMID- 17465504 TI - Hypermethylation and aberrant expression of Wnt antagonist secreted frizzled related protein 1 in gastric cancer. AB - AIM: To identify the methylation of secreted frizzled-related protein 1 (SFRP1) in gastric cancer and to investigate the aberrant expression of SFRP1 and its correlation with the clinical pathological features of patients. METHODS: We determined SFRP1 methylation and SFRP1 mRNA expression in 3 gastric cancer cell lines SGC-7901, BGC-823, HGC-27, from 52 primary gastric cancer specimens and matched tumor adjacent tissue specimens by methylation-specific (MSP) PCR and RT PCR respectively. Fisher's exact test was used to analyze the statistical association between clinical pathological data and aberrant expression of SFRP1. RESULTS: In 3 cancer cell lines, BGC-823 and HGC-27 had methylated SFRP1 and lost SFRP1 mRNA expression. After treatment of BGC-823 and HGC-27 with the demethylating agent, 5-aza-2'-deoxycytidine, SFRP1 was re-expressed. In 52 primary gastric cancer specimens and matched tumor adjacent tissue specimens, hypermethylation of SFRP1 was detected in 23 (44%) and 8 (15%) specimens respectively (chi(2) = 10.34, P < 0.01). Loss of SFRP1 expression was detected in 17(33%) and 6 (12%) specimens respectively (chi(2) = 6.75, P < 0.01). There was a significant correlation between SFRP1 hypermethylation and SFRP1 expression loss. SFRP1 expression was also correlated significantly with tumor stage and lymph node status, but not with patient sex, age and histological type. CONCLUSION: SFRP1 inactivation is a common and early event caused mainly by hypermethylation in gastric cancer. SFRP1 expression loss may be correlated with tumor metastasis in primary gastric cancer. PMID- 17465505 TI - Gene expression analysis of pancreatic cystic neoplasm in SV40Tag transgenic mice model. AB - AIM: To study the gene expression changes in pancreatic cystic neoplasm in SV40Tag transgenic mice model and to provide information about the prevention, clinical diagnosis and therapy of pancreatic cancer. METHODS: Using the pBC SV40Tag transgenic mice model of pancreatic cystic neoplasm, we studied the gene expression changes by applying high-density microarrays. Validation of part gene expression profiling data was performed using real-time PCR. RESULTS: By using high-density oligonucleotide microarray, of 14113 genes, 453 were increased and 760 decreased in pancreatic cystic neoplasm, including oncogenes, cell-cycle related genes, signal transduction-related genes, skeleton-related genes and metabolism-related genes. Among these, we confirmed the changes in Igf, Shh and Wnt signal pathways with real-time PCR. The results of real-time PCR showed similar expression changes in gene chip. CONCLUSION: all the altered expression genes are associated with cell cycle, DNA damage and repair, signal pathway, and metabolism. SV40Tag may cooperate with several proteins in promoting tumorigenesis. PMID- 17465506 TI - Glutamine and recombinant human growth hormone protect intestinal barrier function following portal hypertension surgery. AB - AIM: To evaluate the effects of combined treatment of glutamine (Gln) and recombinant human growth hormone(rhGH) on intestinal barrier function following portal hypertension surgery. METHODS: This study was designed as a prospective, randomized and controlled clinical trial. Forty two patients after portal hypertension surgery were randomly assigned into 2 groups: control group (n = 20) and supplemental group (adding Gln and rhGH, n = 22). Every patient received isocaloric and isonitrogenous standard total parenteral nutrition (TPN) starting 3 d after surgery for 7 d. Blood samples were obtained before surgery and at the 3rd and 10th day postoperatively. Host immunity was evaluated by measuring levels of CD4, CD8, CD4/CD8, IgG, IgM and IgA, and the inflammatory responses were determined by assessing IL-2, TNF-alpha and C-reactive protein (CRP) levels. Intestinal permeability and integrity was evaluated by L/M test and histological examination, respectively. RESULTS: On postoperative d 10, CD4, CD4/CD8, IgG and IL-2 levels in supplemental group were significantly higher than those in control group (33.7 +/- 5.5 vs 31.0 +/- 5.4, P < 0.05, (1.17 +/- 0.32 vs 1.05 +/- 0.15, P < 0.05, 13.94 +/- 1.09 vs 12.33 +/- 1.33, P < 0.05, and 368.12 +/- 59.25 vs 318.12 +/- 45.65, P < 0.05, respectively), whereas the increase in serum TNF alpha concentration was significantly reduced (41.02 +/- 27.56 vs 160.09 +/- 35.17, P < 0.05). The increase in L/M ratio was significantly lower in the supplemental group than in the control group (0.0166 +/- 0.0017 vs 0.0339 +/- 0.0028, P < 0.05). Moreover, mucosal integrity in the supplemental group was better than in the control group. CONCLUSION: Postoperative administration of TPN supplemented with Gln and rhGH in patients after portal hypertension surgery improves immune function, modulates inflammatory response, prevents the intestinal mucous membrane from atrophy and preserves intestinal integrity. PMID- 17465507 TI - Effect of acupuncture at Foot-Yangming Meridian on gastric mucosal blood flow, gastric motility and brain-gut peptide. AB - AIM: To observe the effect of acupuncture at Foot-Yangming Meridian on gastric mucosal blood flow (GMBF), gastric motility and brain-gut peptide. METHODS: Sixty SD rats were randomly divided into 6 groups: normal control group, model group (group with gastric mucosal damage, GMD), Sibai group (with acupuncture at Sibai point + GMD), Tianshu group (with acupuncture at Tianshu point + GMD), Zusanli group (with acupuncture at Zusanli point + GMD) and non-acupoint group (with acupuncture at non-acupoint + GMD). The GMD model group was induced by infusing pure alcohol into gastric cavity. H(2) Gas Clearance Test (HGCT) was used to measure GMBF, the frequency and amplitude of gastric motility were measured by the method of aerocyst, the content of brain-gut peptide in sinus ventriculi and bulbus medullae were detected by radioimmunoassay. RESULTS: Inhibitory effect of the frequency and amplitude of gastric motility were shown in model group, and the rates of frequency and amplitude changes were remarkably different from the normal control group (-19.41 +/- 17.21 vs -4.71 +/- 10.32, P < 0.05; -51.61 +/- 29.02 vs 1.81 +/- 14.12, P < 0.01). In comparison with control group, the GMBF was 0.52 +/- 0.161 mL vs 1.03 +/- 0.255 mL per 100g tissue/min, P < 0.01, the content of motilin in sinus ventriculi and bulbus medullae was 63.04 +/- 7.77 pg/mL vs 72.91 +/- 8.42 pg/mL, P < 0.05 and 50.96 +/- 8.77 pg/mL vs 60.76 +/- 8.05 pg/mL, P < 0.05, but the content of somatostatin in sinus ventriculi and bulbus medullae was 179.85 +/- 43.13 ng/g vs 90.54 +/- 40.42 ng/g, P < 0.01 and 532.86 +/- 122.58 ng/g vs 370.91 +/- 76.29 ng/g, P < 0.05,respectively. In comparison with model group, the amplitude of gastric motility was 1.52 +/- 20.13, -6.52 +/- 23.31, 6.92 +/- 25.21 vs -51.61 +/- 29.02, P < 0.01 and GMBF was 0.694 +/- 0.160 mL vs 0.893 +/- 0.210 mL, 1.038 +/- 0.301 mL vs 0.52 +/- 0.161 mL per 100g tissue/min, P < 0.01, respectively in Tianshu, Sibai and Zusanli groups, the content of motilin in sinus ventriculi and bulbus medullae was 71.64 +/- 9.35 pg/mL vs 63.04 +/- 7.77 pg/mL, P < 0.05; 58.96 +/- 6.69 pg/mL vs 50.96 +/- 8.77 pg/mL, P < 0.05 in Zusanli group and 72.84 +/- 8.62 pg/mL vs 63.04 +/- 7.77 pg/mL, P < 0.05 in Sibai group, while the content of somatostatin in bulbus medullae in Tianshu, Sibai and Zusanli groups was 480.62 +/- 151.69 ng/g, 388.53 +/- 155.04 ng/g, 365.76 +/- 143.93 ng/g vs 532.86 +/- 122.58 ng/g, P < 0.05, respectively. CONCLUSION: Electro-acupuncture at acupoints of Foot-Yangming Meridian could enhance the gastric motility, improve GMBF, and regulate the content of motilin and somatostatin in sinus ventriculi and bulbus. The effects of acupuncture on GMBF and gastric motility may be related to the content of brain-gut peptide. PMID- 17465508 TI - Reversal of multidrug resistance in vincristine-resistant human gastric cancer cell line SGC7901/VCR by LY980503. AB - AIM: To investigate the reversal effect of LY980503, a benflumetol derivative, on multidrug resistance in vincristine (VCR) -resistant human gastric carcinoma cell line SGC7901/VCR. METHODS: Cells of a human gastric cancer cell line, SGC7901, and its VCR-resistant variant, SGC7901/VCR, were cultivated with LY980503 and /or doxorubicin (DOX). The cytotoxicity of drugs in vitro was assayed by MTT method. Based on the flow cytometric technology, the uptake of DOX was detected in these cells by measuring DOX-associated mean fluorescence intensity (MFI). RESULTS: SGC7901/VCR cells were 23.5 times more resistant to DOX in comparison with SGC7901 cells. LY980503 at the concentrations of 2.0 micromol/L-10 micromol/L had no obvious cytotoxicity to SGC7901 and SGC7901/VCR cells. After simultaneous treatment with LY980503 at the concentrations of 2.0, 4.0 and 10 micromol/L, the IC(50) of DOX to SGC7901/VCR cells decreased from 1.6 +/- 0.12 micromol/L to 0.55 +/- 0.024, 0.25 +/- 0.032 and 0.11 +/- 0.015 micromol/L, respectively, thus, increasing the DOX sensitivity by 2.9-fold (P < 0. 05), 6.4-fold (P < 0. 01) and 14.5-fold (P < 0. 01), respectively. In the uptake study of DOX, simultaneous incubation of SGC7901/VCR cells with LY980503 significantly increased the DOX associated MFI in SGC7901/VCR cells. No such results were found in parental SGC7901 cells. CONCLUSION: LY980503 at non-cytotoxic concen-trations can effectively circumvent resistance of SGC7901/VCR cells to DOX by increasing intracellular DOX accumulation. PMID- 17465509 TI - Hepatitis A vaccine associated with autoimmune hepatitis. AB - To describe a case of probable relapsing autoimmune hepatitis associated with vaccination against hepatitis A virus (HAV). A case report and review of literature were written concerning autoimmune hepatitis in association with hepatitis A and other hepatotropic viruses. Soon after the administration of formalin-inactivated hepatitis A vaccine, a man who had recently recovered from an uncharacterized but self-limiting hepatitic illness, experienced a severe deterioration (AST 1687 U/L, INR 1.4). Anti-nuclear antibodies were detectable, and liver biopsy was compatible with autoimmune hepatitis. The observation supports the role of HAV as a trigger of autoimmune hepatitis. Studies in helper T-cell activity and antibody expression against hepatic proteins in the context of hepatitis A infection are summarized, and the concept of molecular mimicry with regard to other forms of viral hepatitis and autoimmunity is briefly explored. PMID- 17465510 TI - Complicated small-bowel diverticulosis: a case report and review of the literature. AB - While jejunoileal diverticula are rare and often asymptomatic, they may lead to chronic non-specific or acute symptoms. The large majority of complications present with an acute abdomen similar to appendicitis, cholecystitis or colonic diverticulitis but they also may appear with atypical symptoms. As a result, diagnosis of complicated jejunoileal diverticulosis can be quite difficult, and may solely depend on the result of surgical exploration. In the absence of contra indications, diagnostic laparoscopy has the benefit of thorough examination of the abdominal contents and helps to reach an absolute diagnosis. Surgical resection of the involved small-bowel segment with primary anastomosis is the preferred treatment in patients with symptomatic complicated jejunoileal diverticular disease. An atypical presentation of complicated jejunal diverticulitis in conjunction with sigmoid diverticulitis diagnosed with laparoscopy and treated with surgical resection is presented. PMID- 17465511 TI - Solitary recurrence of hilar cholangiocarcinoma in a mediastinal lymph node two years after curative resection. AB - We recently encountered an unusual case of hilar cholangiocarcinoma in which a solitary recurrence in a mediastinal lymph node occurred two years after curative resection of the primary tumor. A 64-year old woman was admitted to our hospital with a complaint of right hypochondrial discomfort. After imaging studies demonstrated a hilar cholangiocarcinoma in the left hepatic duct, a curative resection of the tumor was performed, consisting of a left hepatic lobectomy along with caudate lobectomy, regional lymph node dissection, and resection of the extrahepatic bile duct. No nodal metastasis was observed histologically. Two years after surgery, the patient was found to have a nodule in the posterior mediastinum, which was thoracoscopically resected. No other swollen lymph nodes, local recurrence, or distant metastasis were noted. Histologically, the nodule proved to be a metastatic lymph node, and adjuvant chemoradiation therapy was initiated. The patient remained well for the four years following her first operation and had no evidence of disease recurrence 28 mo after her second operation. To our knowledge, this case is the first report of solitary recurrence in a mediastinal lymph node after curative resection of hilar cholangiocarcinoma. PMID- 17465512 TI - Multiple carcinoids in the duodenum, pancreas and stomach accompanied with type A gastritis: a case report. AB - We report a case of multiple duodenal, pancreatic, and gastric carcinoids. A 67 year old woman was admitted to our hospital for treatment of a duodenal carcinoid. Laboratory tests revealed that the patient was associated with macrocytic anemia and hypergastrinemia, and type A gastritis was shown by gastrofiberscopy. During surgery, another tumor was incidentally found in the head of the pancreas. The tumors in the duodenum and pancreas were completely excised by pancreatoduodenectomy and immunohistologically diagnosed as gastrin and serotonin-producing carcinoids, respectively. Pathological examination revealed that in addition to the grossly found carcinoids, there were subclinical carcinoids, one of which was an endocrine cell micronest, located in the stomach and duodenum. The tumors in the duodenum, pancreas, and stomach showed different characteristics from one another morphologically and immunochemically. Although no definitive evidence has been obtained, some sort of genetic anomaly may have been involved in this case, and hypergastrinemia due to duodenal gastrinoma may induce multiple gastric carcinoids. PMID- 17465513 TI - Chemoradiotherapy for a patient with a giant esophageal fistula. AB - We describe our experience of treatment for a giant esophageal malignant fistula, which has not been reported previously. A 36-year-old woman who was diagnosed as having massive esophageal small cell carcinoma with metastases was treated with chemoradiotherapy. However, a giant esophagomediastinal fistula appeared due to shrinkage of the massive tumor, and all anti-cancer treatment was suspended. However, chemoradiotherapy was restarted at the request of the patient despite the presence of the fistula. After restarting treatment, the giant esophageal fistula was naturally closed despite intensive chemoradiotherapy, and the patient became able to eat and drink. Although the patient finally died, her QOL and prognosis seemed to be improved by the chemoradiotherapy. Anti-cancer treatment could be safely performed despite the presence of a giant fistula. The giant fistula closed while intensive chemotherapy was administered to the patient. Therefore, the presence of a fistula may not be a contraindication for curative chemoradiotherapy. Completion of treatment with proper management and maintenance of patients would be of benefit to patients with fistula. PMID- 17465514 TI - Severe delayed complication after percutaneous endoscopic colostomy for chronic intestinal pseudo-obstruction: a case report and review of the literature. AB - Percutaneous endoscopic colostomy (PEC) is increasingly proposed as an alternative to surgery to treat various disorders, including acute colonic pseudo obstruction, chronic intestinal pseudo-obstruction and relapsing sigmoid volvulus. We report on a severe complication that occurred two months after PEC placement. A 74-year-old man with a history of chronic intestinal pseudo obstruction evolving since 8 years was readmitted to our hospital and received PEC to provide long-standing relief. The procedure was uneventful and greatly improved the patient's quality of life. Two months later, the patient developed acute stercoral peritonitis. At laparotomy, the colostomy flange was embedded in the abdominal wall but no pressure necrosis was found at the level of the colonic wall. This complication was likely related to inadvertent traction of the colostomy tube. Subtotal colectomy with terminal ileostomy was performed. We review the major features of 60 cases of PEC reported to date, including indications and complications. PMID- 17465515 TI - Persistent omphalomesenteric duct causing small bowel obstruction in an adult. AB - An extremely rare case of persistent omphalomesenteric duct causing small bowel obstruction is presented. A 20-year-old female patient without medical history presented with colicky abdominal pain, vomiting, absence of passage of gas and feces, and abdominal distension of 24 h duration. Physical examination and blood tests were normal. Abdominal X-ray showed small bowel obstruction. Computed tomography of the abdomen demonstrated dilated small bowel and a band originating from the umbilicus and continuing between the small bowel loops; an omphalomesenteric duct remnant was suspected. In exploratory laparotomy, persistent omphalomesenteric duct causing small bowel obstruction was identified and resected. The patient had an uneventful recovery and was discharged on the 5(th) postoperative day. Although persistent omphalomesenteric duct is an extremely infrequent cause of small bowel obstruction in adult patients, it should be taken into consideration in patients without any previous surgical history. PMID- 17465516 TI - Early detection of response to imatinib therapy for gastrointestinal stromal tumor by using 18F-FDG-positron emission tomography and computed tomography imaging. AB - A 41-year old female with metastatic gastrointestinal stromal tumor was referred to 18F-FDG-positron emission tomography and computed tomography (PET/CT) scan before and after one-month treatment with imatinib (Glivec, Gleevec, Novartis, Basel, Switzerland), a tyrosine kinase inhibitor (400 mg/d). Metabolic response was evaluated before and after one month of therapy. The decrease of the maximum standardised uptake value (SUV) was 79% (from 9.8 to 2.1). Positron emission tomography demonstrated complete metabolic response after one-month of imatinib treatment. Additionally, the previous lesion was compared with the coronal computerized tomographic image. There was no difference in the size of the tumor before and after therapy according to CT images. However, metabolic activity was inhibited. 18F-FDG-PET is a valuable method for the detection of response to one month imatinib treatment in patients with gastrointestinal stromal tumors. PMID- 17465517 TI - A primary intestinal lymphangiectasia patient diagnosed by capsule endoscopy and confirmed at surgery: a case report. AB - Intestinal lymphangiectasia (IL) is a rare disease characterized by dilated lymphatic vessles in the intestinal wall and small bowel mesentery which induce loss of protein and lymphocytes into bowel lumen. Because it most often occurs in the intestine and cannot be detected by upper gastroendoscopy or colonoscopy, and the value of common image examinations such as X-ray and computerized tomography (CT) are limited, the diagnosis of IL is difficult, usually needing the help of surgery. Capsule endoscopy is useful in diagnosing intestinal diseases, such as IL. We here report a case of IL in a female patient who was admitted for the complaint of recurrent edema accompanied with diarrhea and abdominal pain over the last twenty years, and aggravated ten days ago. She was diagnosed by M2A capsule endoscopy as a primary IL and confirmed by surgical and pathological examination. PMID- 17465518 TI - Genetically encoded short peptide tags for orthogonal protein labeling by Sfp and AcpS phosphopantetheinyl transferases. AB - Short peptide tags S6 and A1, each 12 residues in length, were identified from a phage-displayed peptide library as efficient substrates for site-specific protein labeling catalyzed by Sfp and AcpS phosphopantetheinyl transferases (PPTases), respectively. S6 and A1 tags were selected for useful levels of orthogonality in reactivities with the PPTases: the catalytic efficiency, kcat/Km of Sfp-catalyzed S6 serine phosphopantetheinylation was 442-fold greater than that for AcpS. Conversely, the kcat/Km of AcpS-catalyzed A1 labeling was 30-fold higher than that for Sfp-catalyzed A1 labeling. S6 and A1 peptide tags can be fused to N- or C-termini of proteins for orthogonal labeling of target proteins in cell lysates or on live cell surfaces. The development of the orthogonal S6 and A1 tags represents a significant enhancement of PPTase-catalyzed protein labeling, allowing tandem or iterative covalent attachment of small molecules of diverse structures to the target proteins with high efficiency and specificity. PMID- 17465519 TI - A fragment-based approach for the discovery of isoform-specific p38alpha inhibitors. AB - In this study, we describe a novel approach for lead discovery against protein kinases, pharmacophore by interligand nuclear Overhauser effect (ILOE), in which a pair of ligands that bind to adjacent pockets on the target surface is identified by the detection of protein-mediated ILOEs. We demonstrate that a pharmacophore-based search guided by experimental binding data of weakly interacting fragments can be rapidly and efficiently used to identify (or synthesize) high-affinity, selective ligands. Targeting the inactive state of protein kinases represents a promising approach to achieve selectivity and cellular efficacy. In this respect, when we apply the method for the discovery of potent p38alpha inhibitors, we also demonstrate that the resulting bidentate compounds are highly selective and exhibit a cellular activity that parallels their in vitro binding to the inactive form of the kinase. The method is relatively simple and of general applicability, and as such we anticipate its potential implementation against a variety of macromolecular targets, including not only protein kinases but also those involved in protein-protein interactions or even nucleic acids. PMID- 17465520 TI - Inorganic sensing using organofunctional sol-gel materials. AB - This Account describes recent work in the development and applications of sol-gel sensors for concentrated strong acids and bases and metal ions. The use of sol gel films doped with organic indicators for the optical sensing of concentrated strong acids (HCl, 1-10 M) and bases (NaOH, 1-10 M) has been explored, and the development of dual optical sensor approaches for ternary systems (HCl-salt-H 2O and NaOH-alcohol-H 2O) to give acid and salt, as well as base and alcohol, concentrations is discussed. The preparation of transparent, ligand-grafted sol gel monoliths is also described, and their use in the analysis of both metal cations (Cu (2+)) and metal anions [Cr(VI)] is presented. A new model using both metal ion diffusion and immobilization by the ligands in such monoliths has been developed to give metal concentrations using the optical monolith sensors. In addition to optical sensing, a method utilizing ligand-grafted sol-gel films for analyte preconcentration in the electrochemical determination of Cr(VI) has been explored and is discussed. PMID- 17465521 TI - Aromaticity of carbon nanotubes. AB - Carbon nanotubes are composed of cylindrical graphite sheets. Both nanotubes and graphite sheets are benzenoid derivatives composed of sp2 carbon atoms arranged in a hexagonal pattern. Therefore both systems are aromatic. The extent of the aromatic character of a molecule G (here benzenoids) can be explained in terms of the number of possible Kekule structures in G. In this work the Kekule structures in carbon nanotubes and the corresponding, rectangular, graphite-sheets the tubes might originate from, were enumerated. It was shown that (2,2), (3,3), and (4,4) carbon nanotubes are more aromatic than the corresponding, rectangular, planar structures. This explains why it might be more difficult to saturate nanotubes by addition reactions than the respective, "narrow", graphite sheets. PMID- 17465522 TI - Three dissimilarity measures to contrast dendrograms. AB - We discussed three dissimilarity measures between dendrograms defined over the same set, they are triples, partition, and cluster indices. All of them decompose the dendrograms into subsets. In the case of triples and partition indices, these subsets correspond to binary partitions containing some clusters, while in the cluster index, a novel dissimilarity method introduced in this paper, the subsets are exclusively clusters. In chemical applications, the dendrograms gather clusters that contain similarity information of the data set under study. Thereby, the cluster index is the most suitable dissimilarity measure between dendrograms resulting from chemical investigation. An application example of the three measures is shown to remark upon the advantages of the cluster index over the other two methods in similarity studies. Finally, the cluster index is used to measure the differences between five dendrograms obtained when applying five common hierarchical clustering algorithms on a database of 1000 molecules. PMID- 17465523 TI - Conformational coverage by a genetic algorithm: saturation of conformational space. AB - The molecular modeling is traditionally based on analysis of minimum energy conformers. Such simplifying assumptions could doom to failure the modeling studies given the significant variation of the geometric and electronic characteristics across the multitude of energetically reasonable conformers representing the molecules. Moreover, it has been found that the lowest energy conformers of chemicals are not necessarily the active ones with respect to various endpoints. Hence, the selection of active conformers appears to be as important as the selection of molecular descriptors in the modeling process. In this respect, we have developed effective tools for conformational analysis based on a genetic algorithm (GA), published in J. Chem. Inf. Comput. Sci. (1994, 34, 234; 1999, 39 (6), 997) and J. Chem. Inf. Model. (2005, 45 (2), 283). This paper presents a further improvement of the evolutionary algorithm for conformer generation minimizing the sensitivity of conformer distributions from the effect of smoothing parameter and improving the reproducibility of conformer distributions given the nondeterministic character of the genetic algorithm (GA). The ultimate goal of the saturation is to represent the conformational space of chemicals with an optimal number of conformers providing a stable conformational distribution which cannot be further perturbed by the addition of new conformers. The generation of stable conformational distributions of chemicals by a limited number of conformers will improve the adequacy of the subsequent molecular modeling analysis. The impact of the saturation procedure on conformer distributions in a specific structural space is illustrated by selected examples. The effect of the procedure on similarity assessment between chemicals is discussed. PMID- 17465524 TI - Core-shell Au nanoparticle formation with DNA-polymer hybrid coatings using aqueous ATRP. AB - We report here a direct surface-grafting approach to forming DNA-containing polymer shells outside of Au nanoparticles using aqueous atom transfer radical polymerization (ATRP). In this approach, DNA molecules were immobilized on Au particles to introduce ATRP initiators on the surface. The same DNA molecules also acted as particle stabilizers through electrostatic repulsion and allowed particles to stay suspended in water. The immobilized ATRP initiators prompted polymer chain growth under certain conditions to form thick polymer shells outside of the particles. The formation of DNA-polymer hybrids outside of Au nanoparticles was characterized using absorption spectroscopy, dynamic light scattering (DLS), transmission electron microscopy (TEM), and gel electrophoresis. The presence of thick polymer shells improved particle stability in high ionic strength media, whereas particles with the DNA coating only aggregated. A visible color difference between these two particle solutions was clearly observed, providing the basis for DNA sensing in homogeneous solutions. PMID- 17465525 TI - Effect of protein charge on the generation of aggregation-prone conformers. AB - This study describes how charge modification affects aggregation of ovalbumin, thereby distinguishing the role of conformational and electrostatic stability in the process. Ovalbumin variants were engineered using chemical methylation or succinylation to obtain a range of protein net charge from -1 to -26. Charge modification significantly affected the denaturation temperature. From urea induced equilibrium denaturation studies, it followed that unfolding proceeded via an intermediate state. However, the heat-induced denaturation process could still be described as a two-state irreversible unfolding transition, suggesting that the occurrence of an intermediate has no influence on the kinetics of unfolding. By monitoring the aggregation kinetics, the net charge was found not to be discriminative in the process. It is concluded that the dominant factor determining ovalbumin aggregation propensity is the rate of denaturation and not electrostatic repulsive forces. PMID- 17465526 TI - Modeling intrinsic kinetics of enzymatic cellulose hydrolysis. AB - A multistep approach was taken to investigate the intrinsic kinetics of the cellulase enzyme complex as observed with hydrolysis of noncrystalline cellulose (NCC). In the first stage, published initial rate mechanistic models were built and critically evaluated for their performance in predicting time-course kinetics, using the data obtained from enzymatic hydrolysis experiments performed on two substrates: NCC and alpha-cellulose. In the second stage, assessment of the effect of reaction intermediates and products on intrinsic kinetics of enzymatic hydrolysis was performed using NCC hydrolysis experiments, isolating external factors such as mass transfer effects, physical properties of substrate, etc. In the final stage, a comprehensive intrinsic kinetics mechanism was proposed. From batch experiments using NCC, the time-course data on cellulose, cello-oligosaccharides (COS), cellobiose, and glucose were taken and used to estimate the parameters in the kinetic model. The model predictions of NCC, COS, cellobiose, and glucose profiles show a good agreement with experimental data generated from hydrolysis of different initial compositions of substrate (NCC supplemented with COS, cellobiose, and glucose). Finally, sensitivity analysis was performed on each model parameter; this analysis provides some insights into the yield of glucose in the enzymatic hydrolysis. The proposed intrinsic kinetic model parametrized for dilute cellulose systems forms a basis for modeling the complex enzymatic kinetics of cellulose hydrolysis in the presence of limiting factors offered by substrate and enzyme characteristics. PMID- 17465527 TI - A novel technique for loading of paclitaxel-PLGA nanoparticles onto ePTFE vascular grafts. AB - The major cause of hemodialysis vascular access dysfunction (HVAD) is the occurrence of stenosis followed by thrombosis at venous anastomosis sites due to the aggressive development of venous neointimal hyperplasia. Local delivery of antiproliferative drugs may be effective in inhibiting hyperplasia without causing systemic side effects. We have previously demonstrated that paclitaxel coated expanded poly(tetrafluoroethylene) (ePTFE) grafts, by a dipping method, could prevent neointimal hyperplasia and stenosis of arteriovenous (AV) hemodialysis grafts, especially at the graft-venous anastomoses; however, large quntities of initial burst release have remained a problem. To achieve controlled drug release, paclitaxel (Ptx)-loaded poly(lactic-co-glycolic acid) (PLGA) nanoparticles (Ptx-PLGA-NPs) were prepared by the emulsion-solvent evaporation method and then transferred to the luminal surface and inner part of ePTFE vascular grafts through our micro tube pumping and spin penetration techniques. Scanning electron microscope (SEM) images of various stages of Ptx-PLGA-NPs unequivocally showed that micro tube pumping followed by spin penetration effectively transferred Ptx-PLGA-NPs to the inner part, as well as the luminal surface, of an ePTFE graft. In addition, the in vitro release profiles of paclitaxel demonstrated that this new system achieved controlled drug delivery with a reduced initial burst release. These results suggest that loading of Ptx PLGA-NPs to the luminal surface and the inner part of an ePTFE graft is a promising strategy to ultimately inhibit the development of venous neointimal hyperplasia. PMID- 17465528 TI - Monitoring of conductivity changes in passive layers by scanning electrochemical microscopy in feedback mode: localization of pitting precursor sites on surfaces of multimetallic phase materials. AB - Scanning electrochemical microscopy (SECM) in feedback mode was applied to monitor changes in the electronic conductivity of a passive layer existing on a nanocrystalline Nd-Fe-B-type permanent magnet of the following composition, Nd13.5Fe79.5Si1B6 (where subscripts denote at. %). This model magnet being a complex multimetallic-phase material (fabricated according to the hydrogenation disproportionation-desorption-recombination route) contained intrinsic iron inclusions, as evident from SEM and EDX mapping. SECM measurements performed in feedback mode using ferrocenecarboxylic acid as a mediator (in phosphate buffer at pH 7) in the presence and absence of chloride anions permitted evaluation of the local surface (passive layer) conductivity changes during pitting corrosion caused by Cl- anions. The spots of high conductivity (electronic or redox) act as precursor sites for initiation of pits. It can be rationalized that iron inclusions are responsible for the high susceptibility of Nd-Fe-B magnets to pitting corrosion. The approach is fairly general and allows localization of pitting precursor sites under different corrosion environments that include a wide range of concentrations of anions causing the passive layer breakdown. PMID- 17465529 TI - Spatiotemporal micropatterning of cells on arbitrary substrates. AB - This paper presents a readily accessible patterning platform--based upon geometric constraints, discrete cell suspension droplets, and controlled cell settling--that provides both temporal and spatial patterning capabilities. As a demonstration, single-cell (and bead) suspensions as well as multicellular human embryonic stem cell colonies are spatiotemporally patterned onto arbitrary substrates. These substrates include tissue culture surfaces, cell monolayers, protein-coated surfaces, and 3D gel matrices. The generation of soluble factor gradients is also demonstrated. This method is completely passive and does not require external power sources. Spatiotemporal patterning provides a foundation for future biological studies that explore the time-dependent relationships between cell-cell signaling and cellular responses. PMID- 17465530 TI - Time-of-flight secondary ion mass spectrometric analysis of polymer tertiary structure in Langmuir monolayer films of poly(dimethylsiloxane). AB - A series of model systems of poly(dimethylsiloxane) (PDMS) of molecular mass 2400 Da and low polydispersity (1.09) were prepared using the Langmuir-Blodgett technique to investigate the effects of tertiary structure on the ion formation probability in time-of-flight secondary ion mass spectrometry (TOF-SIMS). Using data from the measured surface pressure-area isotherm for PDMS ordered monolayer, films were transferred to silver mirror substrates from the various regions in the isotherm where structural changes are observed. Two particular conformations of the polymer are examined here: a linear caterpillar-like structure and a coiled helical structure. Reflection absorption Fourier transform infrared spectroscopic analysis indicates structural changes for the films related to adjustments to the orientation of the polymeric backbone consistent with the formation of two distinct structures. The polymeric changes translate into differences in ion formation probability of fragments in both the low (<250 Da) and high (>1000 Da) mass range. Data are also presented to analyze how tertiary structure may affect the apparent polydispersity index calculated from the TOF SIMS spectra. PMID- 17465531 TI - Surface-enhanced Raman scattering based nonfluorescent probe for multiplex DNA detection. AB - To provide rapid and accurate detection of DNA markers in a straightforward, inexpensive, and multiplex format, an alternative surface-enhanced Raman scattering based probe was designed and fabricated to covalently attach both DNA probing sequence and nonfluorescent Raman tags to the surface of gold nanoparticles (DNA-AuP-RTag). The intensity of Raman signal of the probes could be controlled through the surface coverage of the nonfluorescent Raman tags (RTags). Detection sensitivity of these probes could be optimized by fine-tuning the amount of DNA molecules and RTags on the probes. Long-term stability of the DNA-AuP-RTag probes was found to be good (over 3 months). Excellent multiplexing capability of the DNA-AuP-RTag scheme was demonstrated by simultaneous identification of up to eight probes in a mixture. Detection of hybridization of single-stranded DNA to its complementary targets was successfully accomplished with a long-term goal to use nonfluorescent RTags in a Raman-based DNA microarray platform. PMID- 17465532 TI - Determination of OH number densities outside of a platinum catalyst using cavity ringdown spectroscopy. AB - It is demonstrated that cavity ringdown spectroscopy (CRDS) can be used to probe reaction intermediates desorbing from the surface during a heterogeneous catalytic reaction and provide information valuable in understanding the reaction kinetics. During water formation from H2 and O2, desorbed OH molecules outside of a polycrystalline platinum catalyst were quantified as a function of the relative hydrogen concentration, alphaH2 using CRDS. The temperature of the catalyst was 1500 K, the total pressure was 26 Pa, and the flow was set to 100 sccm. At a distance of 6.5 mm from the Pt catalyst, the maximum OH concentration was found to be 1.5+/-0.2x10(12) cm(-3) at an alphaH2 value of 10%, and the rotational temperature was determined to be 775+/-24 K. The desorbed OH molecules were also probed using laser-induced fluorescence (LIF), and the alphaH2-dependent OH abundance was compared with the CRDS results. The relative concentration of OH probed with LIF appeared to be lower at alphaH2=30-50% compared to what was determined by CRDS. The observed discrepancy is suggested to be due to electronic quenching, as was indicated by a shorter fluorescence lifetime at alphaH2=30% compared to at alphaH2=10%. PMID- 17465533 TI - Infrared-infrared double resonance spectroscopy of the isomers of acetylene-HCN and cyanoacetylene-HCN in helium nanodroplets. AB - Infrared-infrared double resonance spectroscopy is used to probe the vibrational dynamics of molecular complexes solvated in helium nanodroplets. We report results for the acetylene-HCN and cyanoacetylene-HCN binary complexes, each having two stable isomers. We find that vibrational excitation of an acetylene HCN complex results in a population transfer to the other isomer. Photoinduced isomerization is found to be dependent on both the initially excited vibrational mode and the identity of the acetylene-HCN isomer. However, population transfer is not observed for the cyanoacetylene-HCN complexes. The results are rationalized in terms of the ab initio intermolecular potential energy surfaces for the two systems with particular emphasis on the long-range barriers to rearrangement. PMID- 17465534 TI - Two liquid phases of water in the deeply supercooled region and their roles in crystallization and formation of LiCl solution. AB - The properties of supercooled liquid water and the mechanism of crystallization in it were investigated using time-of-flight secondary ion mass spectrometry and reflection absorption infrared spectroscopy. The self-diffusion of the water molecules commences at 136 K, and then the liquid-liquid phase transition occurs at 160-165 K. The latter is evidenced not only by the occurrence of fluidity but also by the formation of a LiCl solution. The infrared absorption band also changes drastically above 160 K due to crystallization of water (on the Au film) and the formation of LiCl solution (on the LiCl film). The immediate crystallization and dissolution of LiCl are thought to be characteristic of normal water that is created in a deeply supercooled region, indicating that viscous liquid water (T > 136 K) is transformed into supercooled liquid water at around 160 K. The crystallization kinetics is different between these two phases because the former (latter) involves nuclear growth (spontaneous nucleation). Without nuclei, crystallization is quenched below 160 K in the present experiment. It is suggested that the viscous liquid phase coexists at the surface or grain boundaries of metastable ice Ic. PMID- 17465535 TI - Charging and aggregation of positively charged latex particles in the presence of anionic polyelectrolytes. AB - Charging behavior and colloidal stability of amidine latex particles are studied in the presence of poly(sodium styrene sulfonate) (PSS) and KCl. Detailed measurements of electrophoretic mobility, adsorbed layer thickness, and aggregation (or coagulation) rate constant on varying the polymer dose, molecular mass of the polymer, and ionic strength are reported. Polyelectrolyte adsorption leads to the characteristic charge reversal (or overcharging) of the colloidal particles at the isoelectric point (IEP). In accordance with classical Derjaguin Landau-Verwey-Overbeek (DLVO) theory, uncharged particles tend to aggregate because of van der Waals attraction, whereas charged particles are stabilized by electrical double layer repulsion. Attractive patch-charge interactions originating from the laterally inhomogeneous structure of the adsorbed polymer substantially decrease the suspension stability or even accelerate the aggregation rate beyond diffusion control. These electrostatic non-DLVO forces become progressively important with increasing molecular mass of the polymer and the ionic strength of the solution. At higher polymer dose of typically 10 times the IEP, one observes the formation of a saturated layer of the adsorbed polymer with a thickness of several nanometers. Its thickness increases with increasing molecular mass, whereby the layer becomes increasingly porous. This layer does not seem to be involved in the suspension stabilization, since at such high polymer doses the double layer repulsion has attained sufficient strength to stabilize the suspension. PMID- 17465536 TI - Backbone motions of free and pheromone-bound major urinary protein I studied by molecular dynamics simulation. AB - Molecular motions of free and pheromone-bound mouse major urinary protein I, previously investigated by NMR relaxation, were simulated in 30 ns molecular dynamics (MD) runs. The backbone flexibility was described in terms of order parameters and correlation times, commonly used in the NMR relaxation analysis. Special attention was paid to the effect of conformational changes on the nanosecond time scale. Time-dependent order parameters were determined in order to separate motions occurring on different time scales. As an alternative approach, slow conformational changes were identified from the backbone torsion angle variances, and "conformationally filtered" order parameters were calculated for well-defined conformation states. A comparison of the data obtained for the free and pheromone-bound protein showed that some residues are more rigid in the bound form, but a larger portion of the protein becomes more flexible upon the pheromone binding. This finding is in general agreement with the NMR results. The higher flexibility observed on the fast (fs-ps) time scale was typically observed for the residues exhibiting higher conformational freedom on the ns time scale. An inspection of the hydrogen bond network provided a structural explanation for the flexibility differences between the free and pheromone-bound proteins in the simulations. PMID- 17465537 TI - Why does Kevlar decompose, while Nomex does not, when treated with aqueous chlorine solutions? AB - Kevlar and Nomex are high-performance polymers which have wide varieties of applications in daily life. Recently, they have been proposed to be biocidal materials when reacted with household bleach (sodium hypochlorite solution) because they contain amide moieties which can be chlorinated to generate biocidal N-halamine functional groups. Although Nomex can be chlorinated without any significant decomposition, Kevlar decomposes under the same chlorination conditions. In this study, two mimics for each of the polymers were synthesized to simulate the carboxylate and diaminophenylene components of the materials. It was found that the p-diaminophenylene component of the Kevlar mimic is oxidized to a quinone-type structure upon treatment with hypochlorous acid, which then decomposes. However, such a mechanism for the Nomex mimic is not possible. In this paper, based upon these observations, a plausible answer will be provided to the title question. PMID- 17465539 TI - New BEDT-TTF/[Fe(C5O5)3]3- hybrid system: synthesis, crystal structure, and physical properties of a chirality-induced alpha phase and a novel magnetic molecular metal. AB - The paramagnetic and chiral anion [Fe(C5O5)3]3- (C5O52-=croconate) has been combined with the organic donor BEDT-TTF (=ET=bis(ethylenedithio)tetrathiafulvalene) to synthesize a novel paramagnetic semiconductor with the first chirality-induced alpha phase, alpha-(BEDT TTF)5[Fe(C5O5)3].5H2O (1), and one of the few known paramagnetic molecular metals, beta-(BEDT-TTF)5[Fe(C5O5)3].C6H5CN (2). Both compounds present layers of BEDT-TTF molecules, with the alpha or beta packing modes, alternating with layers containing the high-spin S=5/2 Fe(III) anions and solvent molecules. In the alpha phase, the alternation of the chiral [Fe(C5O5)3]3- anions along the direction perpendicular to the BEDT-TTF chains induces an alternation of the tilt angle of the BEDT-TTF molecules, giving rise to the observed alpha phase. The alpha phase presents a semiconductor behavior with a high room-temperature conductivity (6 S.cm-1) and an activation energy of 116 meV. The beta phase presents a metallic behavior down to ca. 120 K, where a charge localization takes place with a reentrance to the metallic state below ca. 20 K followed by a metal-semiconductor transition at ca. 10 K. The magnetic properties are dominated by the paramagnetic S=5/2 [Fe(C5O5)3]3- anion with an extra Pauli-type paramagnetism in the metallic beta phase. The ESR spectra confirm the presence of the high-spin Fe(III) containing anion and show a progressive localization in the organic sublattice along with an antiferromagnetic coupling below ca. 120 K that, in the metallic beta phase, could be at the origin of the transition from the metallic to the activated conductivity regime. The correlation between crystal structure and conductivity behavior has been studied by means of tight-binding band structure calculations which provide a rationalization of the charge distribution and conductivity results. PMID- 17465540 TI - Metalation of cyclic pseudopeptidic thiosulfinates with Ni(II) and Zn(II) after ring opening: a mechanistic investigation. AB - Thiosulfinates are an emerging class of oxidized sulfur species that are frequently supposed to be involved in biochemical processes. Reaction of 12- and 10-membered ring pseudopeptidic thiosulfinates 1a (4,4,7,7-tetramethyl 1,3,4,7,8,10-hexahydro-5,6,1,10-benzodithiadiazacyclododecine-2,9-dione 5-oxide) and 1b (3,3,6,6-tetramethyl-1,8-dihydro-4,5,1,8-benzodithiadiazecine-2,7(3H,6H) dione 4-oxide) with a Ni(II) salt leads after ring cleavage under alkaline conditions to the isolation of diamidato/thiolato/sulfinato complexes. These two thiolato/sulfinato complexes of nickel, which can also be prepared by dioxygen oxidation of the parent diamidato/dithiolato complexes, were characterized by X ray crystallography. They show a square-planar geometry with a S-bonded sulfinato ligand. A similar reaction between 1b and a Zn(II) salt leads to a thiolato/sulfinato complex with an O-bonded sulfinate via the intermediate formation of a mixed thiolato/sulfinic ester. On the basis of 1H NMR, IR, and mass analyses, the sulfinic ester in the intermediate is proposed to be O-bonded to the zinc center. Then, an in-depth study of the cleavage of these thiosulfinates with the oxyanions RO- and HO- was performed. This led, after trapping of the open species with CH3I, to the identification of three polyfunctionalized products containing a methyl thioether, with either an isothiazolidin-3-one S-oxide, a methyl sulfone, or a methyl sulfinic ester. All of these products arise from a selective nucleophilic attack at the sulfinyl sulfur, promoted either directly by RO- or HO- or by an internal peptidic nitrogen of the thiosulfinate after deprotonation with RO- or HO-. PMID- 17465541 TI - Synthesis and structures of Mo3Se7Te2Br10, Mo3Se7TeI6, and Mo6Te21I22 containing TeX3- (X=Br, I) ligands coordinated to a triangular cluster core. AB - New ternary and quaternary molybdenum cluster chalcohalides were obtained by high temperature reactions between Mo, chalcogens, and halogens in evacuated ampules. The crystal structures of [Mo3Se7(TeBr3)Br2]2[Te2Br10] (1), [Mo3Se7(TeI3)I2]I (2), and [Mo3Te7(TeI3)3]2(I)(TeI3) (3) were determined by single-crystal X-ray diffraction. The structures of 1 and 2 consist of positively charged zigzag chains infinity1 [Mo3Se7(TeX3)X4/2] (X=Br, I), with Te2Br102- and I-, respectively, as counterions. The TeI3- and TeBr3- ions function as bidentate ligands in 1 and 2. In 3, TeI3- is not coordinated to the metal but acts as a counterion to the [Mo3Te7(TeI3)3]+ cluster cation. PMID- 17465542 TI - Zero to three dimensional increase of silver(I) coordination assemblies controlled by deprotonation of 1,3,5-tri(2-benzimidazolyl)benzene and aggregation of multinuclear building units. AB - Four Ag(I) complexes of a triangular multidentate ligand 1,3,5-tri(2 benzimidazolyl)benzene (H3TBimB), namely, [Ag2(H3TBimB)2](CF3SO3)2 (1), [Ag4(HTBimB)2]n (2), [Ag9(HTBimB)4(TAZ)]n (HTAZ=1,2,4-triazole) (3), and [Ag17(TBimB)5(HTBimB)(H2O)5]n.nH2O (4), have been synthesized at different pH values adjusted by addition of NH3.H2O under solvothermal conditions and characterized by X-ray single-crystal diffraction. Complex 1 shows an M2L2 dimeric structure, 2 displays a one-dimensional chain containing M4L2 basic units, 3 is a two-dimensional network built up from an M9L4 subunit, and 4 exhibits a three-dimensional framework generated by an M17L6 motif. Dimensional increase in complexes 1-4 was caused by deprotonation of the H3TBimB ligand, thus offering more coordinating donors and resulting in aggregation of oligomeric Ag(I) building units. In the cases of complexes 3 and 4, TAZ or H2O molecules serve as auxiliary ligands to complete the coordination geometry of the Ag(I) ions wherever necessary. The photoluminescent properties of the ligand H3TBimB and the complexes 1-3 have been investigated. PMID- 17465543 TI - Adsorption energy, growth mode, and sticking probability of Ca on poly(methyl methacrylate) surfaces with and without electron damage. AB - The adsorption of Ca atoms on pristine and electron-irradiated poly(methyl methacrylate) (PMMA) surfaces at 300 K has been studied by adsorption microcalorimetry, atomic beam/surface scattering, and low-energy He+ ion scattering spectroscopy (ISS). On pristine PMMA, the initial sticking probability of Ca is 0.5, increasing quickly with Ca coverage. Below 0.5 ML, the heat of adsorption is 730-780 kJ/mol, much higher than Ca's sublimation energy (178 kJ/mol). The Ca here is invisible to ISS, which is attributed to Ca binding to ester groups below the CH3/CH2-terminated PMMA surface. The adsorption energy increases with coverage, suggesting attractions between neighboring Ca-ester complexes. Above 0.5 ML, Ca starts to grow as three-dimensional (3D) Ca clusters on top of the surface, which dominate growth after 2 ML. It is proposed that each Ca reacts with two esters to form the Ca carboxylate of PMMA, because this reaction's heat would be close to that observed. The total amount of Ca that binds to subsurface sites is estimated from the integral heat of adsorption to involve 4-6 layers of ester groups. Exposing the PMMA surface to electrons increases Ca's initial sticking probability but lowers its adsorption energy. This is attributed to electron-induced defects acting as nucleation sites for 3D Ca islands, whose growth now competes kinetically with Ca diffusing to subsurface esters. Consequently, only two layers of subsurface esters get populated at saturation. The heat eventually reaches Ca's bulk heat of sublimation on all PMMA surfaces, where pure, bulk-like Ca thin films form. PMID- 17465544 TI - A complete macroion-"blackberry" assembly-macroion transition with continuously adjustable assembly sizes in {Mo132} water/acetone systems. AB - A complete, continuous transition from discrete macroions to blackberry structures, and then back to discrete macroions, is reported for the first time in the system of {Mo132}/water/acetone, with {Mo132} (full formula (NH4)42[Mo132O372(CH3COO)30(H2O)72].ca.300H2O.ca.10CH3COONH4) as the C60-like anionic polyoxomolybdate molecular clusters. Laser light scattering studies reveal the presence of the self-assembled {Mo132} blackberry structures in water/acetone mixed solvents containing 3 vol % to 70 vol % acetone, with the average hydrodynamic radius (Rh) of blackberries ranging from 45 to 100 nm with increasing acetone content. Only discrete {Mo132} clusters are found in solutions containing <3 vol % and >70 vol % acetone. The complete discrete macroion (cluster)-blackberry-discrete macroion transition helps to identify the driving forces behind the blackberry formation, a new type of self-assembly process. The charge density on the macroions is found to greatly affect the blackberry formation and dissociation, as the counterion association is very dominant around blackberries. The transitions between single {Mo132} clusters and blackberries, and between the blackberries with different sizes, are achieved by only changing the solvent quality. PMID- 17465545 TI - Infrared characterization of hydroxyl groups on MgO: a periodic and cluster density functional theory study. AB - The infrared OH stretching frequencies of the various types of hydroxyl groups on MgO surfaces have been calculated by periodic (VASP) and cluster (Gaussian) DFT simulations. Surface irregularities (mono and diatomic steps, corners, step divacancies, and kinks) have been considered to model the IR spectra of hydroxylated MgO powders. A good correspondence between calculated and experimental frequencies is obtained with the B3LYP functional. Hydrogen-bonding is the parameter which influences most the IR frequency of OH groups, followed by location of OH groups in concave or convex areas of the surface and then oxygen coordination. The evolution of experimental IR spectra upon evacuation at increasing temperature can be rationalized on the basis of calculated thermal stabilities of each kind of OH groups. A new model is finally proposed to help assign the experimental bands, in terms of hydrogen-bonding, local topology of the hydroxylated sites, and coordination of oxygen. PMID- 17465546 TI - Enhancing one-dimensional charge transport through intermolecular pi-electron delocalization: conductivity improvement for organic nanobelts. PMID- 17465547 TI - Carbon dioxide fixation into chemicals (methyl formate) at high yields by surface coupling over a Pd/Cu/ZnO nanocatalyst. PMID- 17465548 TI - Strong intermolecular electronic coupling within a tetrathiafulvalene island embedded in self-assembled monolayers. AB - Electroactive tetrathiafulvalene thiol, specially designed to pursue an intermolecular electronic coupling, was embedded in an n-alkanethiol SAM matrix as islands and was studied under potential control using in situ scanning tunneling microscopy. The apparent height of the islands increased with the island size, irrespective of the oxidation state of the tetrathiafulvalene backbones. This behavior can be rationalized on the basis of the strong intermolecular electronic coupling that creates efficient intermolecular conduction paths. PMID- 17465549 TI - Crucial role of anions on the deprotonation of the cationic dihydrogen complex trans-[FeH(eta2-H2)(dppe)2]+. AB - The kinetics of reaction of the dihydrogen complex trans-[FeH(eta2-H2)(dppe)2]+ with an excess of NEt3 to form cis-[FeH2(dppe)2] shows a first-order dependence with respect to both the metal complex and the base. The corresponding second order rate constant only shows minor changes when the solvent is changed from THF to acetone. However, the presence of salts containing the BF4-, PF6-, and BPh4- anions causes larger kinetic changes, the reaction being accelerated by BF4- and PF6- and decelerated in the presence of BPh4-. These results can be interpreted considering that the ion pairs formed by the complex and the anion provide a reaction pathway more efficient than that going through the unpaired metal complex. From the kinetic results in acetone solution, the stability of the ion pairs and the rate constant for their conversion to the reaction products have been derived. Theoretical calculations provide additional information about the reaction mechanism both in the absence and in the presence of anions. In all cases, the reaction occurs with proton transfer from the trans-dihydride to the base through intermediate structures showing Fe-H2...N and Fe-H...H...N dihydrogen bonds, isomerization to the cis product occurring once the proton transfer step has been completed. Optimized geometries for the ion pairs show that the anions are placed close to the H2 ligand. In the case of BPh4-, the bulky phenyls hinder the approach of the base and make the ion pairs unproductive for proton transfer. However, ion pairs with BF4- and PF6- can interact with the base and evolve to the final products, the anion accompanying the proton through the whole proton transfer process, which occurs with an activation barrier lower than for the unpaired metal complex. PMID- 17465550 TI - Understanding the effect of carbonate ion on cisplatin binding to DNA. PMID- 17465551 TI - Synthesis of molecular brushes by "grafting onto" method: combination of ATRP and click reactions. AB - Molecular brushes (densely grafted polymers or bottle-brush macromolecules) were synthesized by the "grafting onto" method via combination of atom transfer radical polymerization (ATRP) and "click" reactions. Linear poly(2-hydroxyethyl methacrylate) (PHEMA) polymers were synthesized first by ATRP. After esterification reactions between pentynoic acid and the hydroxyl side groups, polymeric backbones with alkynyl side groups on essentially every monomer unit (PHEMA-alkyne) were obtained. Five kinds of azido-terminated polymeric side chains (SCs) with different chemical compositions and molecular weights were used, including poly(ethylene glycol)-N3 (PEO-N3), polystyrene-N3, poly(n-butyl acrylate)-N3, and poly(n-butyl acrylate)-b-polystyrene-N3. All click coupling reactions between alkyne-containing polymeric backbones (PHEMA-alkyne) and azido terminated polymeric SCs were completed within 3 h. The grafting density of the obtained molecular brushes was affected by several factors, including the molecular weights and the chemical structures of the linear SCs, as well as the initial molar ratio of linear chains to alkynyl groups. When linear polymers with "thinner" structure and lower molecular weight, e.g., PEO-N3 with Mn = 775 g/mol, were reacted with PHEMA-alkyne (degree of polymerization = 210) at a high molar ratio of linear chains to alkynyl groups in the backbone, the brush copolymers with the highest grafting density were obtained (Y(grafting) = 88%). This result indicates that the average number of SCs was ca. 186 per brush molecule and the average molecular weight of the brush molecules was ca. 190 kg/mol. PMID- 17465552 TI - Discrete heterogeneous quaternary structure formed by alpha/beta-peptide foldamers and alpha-peptides. PMID- 17465553 TI - Magnetic tweezers measurement of the bond lifetime-force behavior of the IgG protein A specific molecular interaction. AB - The bond lifetime-force behavior of the immunoglobulin G (IgG)-protein A interaction has been studied with magnetic tweezers to characterize the physical properties of the bond under nonequilibrium conditions. Super-paramagnetic microparticles were developed that have a high and uniform magnetization to simultaneously apply a piconewton-scale tensile force to many thousands of IgG protein A bonds. A strong and a weak slip bond were detected with an effective bond length that is characteristic of short-range, stiff intermolecular interactions. These bonds are attributed to the interaction of protein A with the constant region (Fc) and heavy chain variable domain (VH) of IgG, respectively. The IgG-VH interaction appears to be one of the weakest specific molecular interactions that has been identified with a single molecule force measurement technique. This study demonstrates that magnetic tweezers can be used to rapidly characterize very weak biomolecular interactions as well as strong biomolecular interactions with a high degree of accuracy. PMID- 17465554 TI - Laser photoexcitation of NAD(P)H induces reduction of P450 BM3 heme domain on the microsecond time scale. AB - We demonstrate that photoexcitation of NAD(P)H at 355 nm using a Nd:YAG laser leads to rapid reduction of the heme domain of the Bacillus megaterium fatty acid hydroxylase flavocytochrome P450 BM3. An aqueous electron derived from photoexcited NAD(P)H is rapidly transferred to the heme domain, enabling the formation of a carbon monoxy complex of the ferrous P450 (FeII-CO) on the microsecond time scale. Using this approach we have determined the limiting rate constant (1770 s-1 for substrate-free heme domain) for formation of the FeII-CO complex. We find no dependence of the observed rate of FeII-CO complex formation on NAD(P)H concentration but demonstrate a hyperbolic dependence on carbon monoxide concentration. The apparent dissociation constant for the complex of carbon monoxide bound noncovalently to the ferric form of the BM3 heme domain (and with NADH as reductant) is 323 microM. Binding of a P450 substrate (N palmitoylglycine) weakened the complex between carbon monoxide and the ferric BM3 heme domain (Kd increased to 1404 microM) but enhanced the rate of formation of the FeII-CO complex (3036 s-1 for substrate-free heme domain). This study demonstrates the applicability of NAD(P)H photoexcitation as a method for rapid electron delivery to P450 enzymes and provides a new route to probing the P450 catalytic cycle and its transient intermediates. PMID- 17465557 TI - Biomimetic synthesis of the tetracyclic core of berkelic acid. AB - Acid-catalyzed condensation of 2,6-dihydroxybenzoic acid 3 with ketal aldehyde 14 in methanol at 25 degrees C, followed by CH2N2 esterification, gave a 4:1:4:1 mixture of diastereomers 15b-18b in 60% yield. Equilibration of this mixture with TFA in CDCl3 provided tetracycle 15b (83% yield) with the complete skeleton of berkelic acid. A similar condensation at 0 degrees C afforded 15b-18b and a reduction product 19b, which was probably formed by a 1,5-hydride shift. PMID- 17465558 TI - Aminals as substrates for sulfur ylides: a synthesis of functionalized aziridines and N-heterocycles. AB - Sulfur ylides stabilized by Ar, vinyl, or amide groups react with five-membered ring tert-butylsulfinyl aminals to give functionalized chiral, nonracemic aziridines in high yield and with good selectivities (up to 15:1 trans:cis, up to >95:5 trans dr, always >95:5 cis dr). The intermediate aziridines can be converted into pyrrolidines or piperidines depending on the reaction conditions. PMID- 17465559 TI - Rhodium-catalyzed conjugate addition-enantioselective protonation: the synthesis of alpha,alpha'-dibenzyl esters. AB - Alpha-benzyl acrylates, which are conveniently prepared from the corresponding aldehydes, can be employed as substrates in a tandem rhodium-catalyzed conjugate addition-enantioselective protonation protocol to afford enantiomerically enriched alpha,alpha'-dibenzyl esters. The synergistic effect of enantiopure ligand and proton source was rapidly optimized with use of a microwave reactor. PMID- 17465560 TI - Rapid synthesis of the A-E fragment of ciguatoxin CTX3C. AB - The A-E fragment of the marine natural product CTX3C has been prepared in an efficient manner by using a strategy in which two-directional and iterative ring closing metathesis (RCM) reactions were employed for ring construction. PMID- 17465561 TI - Gold-catalyzed efficient preparation of linear alpha-iodoenones from propargylic acetates. AB - Only 2 mol % of Au(PPh3)NTf2 is needed to convert readily accessible propargylic acetates into versatile linear alpha-iodoenones in good to excellent yields. This reaction is easy to execute and has broad substrate scope. Good to excellent Z selectivities are observed in the cases of aliphatic propargylic acetates derived from aldehydes. PMID- 17465562 TI - Gold-catalyzed cyclization of allene-substituted malonate esters: synthesis of beta,gamma-unsaturated delta-lactones. AB - An efficient method for the preparation of beta,gamma-unsaturated delta-lactones has been developed. The starting materials for the synthesis of these compounds are allene-substituted malonates which undergo gold-catalyzed cyclization by means of nucleophilic attack of the ester moiety on the allene. It is worth mentioning that this is the first example where an ester group attacks as a nucleophile in a gold-catalyzed transformation of allenes. PMID- 17465563 TI - Control of peptide helix sense by temperature tuning of noncovalent chiral domino effect. AB - We have investigated temperature effect on control of a peptide helix sense through the noncovalent chiral domino effect (NCDE: Inai, Y. et al., J. Am. Chem. Soc. 2003, 125, 8151-8162). Nonapeptide (1: Inai, Y.; Komori, H. Biomacromolecules 2004, 5, 1231-1240), which alone prefers a right-handed helix, maintained a screw-sense balance or a small imbalance at room temperature in the presence of Boc-d-amino acid. Cooling of the solution induced a left-handed helix more clearly. Conversely, heating from room temperature recovered the original right-handed sense. This helix-helix transition was essentially reversible in cooling-heating cycles. An increase in the Boc-d-amino acid concentration elevated temperature for switching CD signs based on the conformational transition. A similar thermal-driven inversion of helix sense was observed for 1 at other initial concentrations, suggesting that this behavior is insensitive to some peptide aggregation. NMR study provided direct evidence for the domino-type control of helix sense, in which Boc-Leu-OH is mainly located at the N-terminal segment. In addition, a left-handed helix induced by the d-isomer was shown to participate in equilibrium with a right-handed helix, whereas the right-handed helix was predominant in the presence of l-isomer. Consequently, we here have proposed a model for controlling a peptide helix sense (or its screw-sense bias) through temperature tuning of the external chiral interaction specific to the N terminal sequence. PMID- 17465564 TI - Investigation of the regioselectivity for the staudinger reaction and its application for the synthesis of aminoglycosides with N-1 modification. AB - The criteria for controlling the regioselectivity of Staudinger reduction of azides have been investigated. These findings enable a convenient direct N-1 modification of the perazidoneamine and perazidoribostamycin resulting in the synthesis of aminoglycoside antibiotics with activity against drug-resistant bacteria. PMID- 17465565 TI - Guanidine/Pd(OAc)2-catalyzed room temperature Suzuki cross-coupling reaction in aqueous media under aerobic conditions. AB - A highly efficient Pd(OAc)2/guanidine aqueous system for the room temperature Suzuki cross-coupling reaction has been developed. The new water-soluble and air stable catalyst Pd(OAc)2.(1f)2 from Pd(OAc)2 and 1,1,3,3-tetramethyl-2-n butylguanidine (1f) was synthesized and characterized by X-ray crystallography. In the presence of Pd(OAc)2.(1f)2, coupling of arylboronic acids with a wide range of aryl halides, including aryl iodides, aryl bromides, even activated aryl chlorides, was carried out smoothly in aqueous solvent to afford the cross coupling products in good to excellent yields and high turnover numbers (TONs) (TONs up to 850,000 for the reaction of 1-iodo-4-nitrobenzene and phenylboronic acid). Furthermore, this mild protocol could tolerate a broad range of functional groups. PMID- 17465566 TI - Convenient route to enantiopure fmoc-protected morpholine-3-carboxylic acid. AB - Enantiopure Fmoc-protected morpholine-3-carboxylic acid was synthesized from dimethoxyacetaldehyde and serine methyl ester through a short and practical synthetic route. The preparation consisted of a five-step process based on reductive amination, intramolecular acetalization, and concomitant elimination of the anomeric methoxy substituent, followed by hydrogenation of the double bond and final acidic ester hydrolysis. The optical purity of both enantiomers of the title amino acid was demonstrated by HPLC analysis of the corresponding amide derivatives obtained from coupling with chiral (S)-(-)-1-phenylethylamine. Moreover, the synthesis of a model tripeptide showed full compatibility of the title Fmoc-amino acid with solid-phase peptide synthesis, thus allowing the application of Fmoc-morpholine-3-carboxylic acid in peptidomimetic chemistry on the solid phase. PMID- 17465567 TI - Solution structure of some lambda(3) Iodanes: an (17)O NMR and DFT study. AB - The structure of a series of I-O bonded bis(acyloxy)iodoarenes and benzoiodoxolones in chloroform solution has been investigated by 17O NMR spectroscopy and by density functional theory (DFT) calculations, employing the PBE0 functional together with the LANL2DZ basis set extended with polarization (d) and diffuse (p) functions. This combined approach allowed us to ascertain that, although these classes of lambda(3) iodanes maintain in chloroform solution their solid state "T-shaped" structure, a degenerate [1,3] sigmatropic shift of iodine between the two oxygens of the acyloxy groups occurs in solution. The energy barrier involved in this process differs in the two classes, thus causing significant differences in the 17O NMR spectra, at room temperature, of the two classes of compounds. PMID- 17465568 TI - First O-glycosylation of hydroxamic acids. AB - The first O-glycosylation of hydroxamic acids is reported. This process involves the use of glycosyl N-phenyl trifluoroacetimidates as glycosyl donors in the presence TMSOTf and 4 A molecular sieves in dichloromethane. Under such conditions, a wide range of new glycosyl donors including glucosyl, galactosyl, mannosyl, glucuronyl, and ribosyl hydroxamates were prepared in good to high yields. This procedure appears to be an advantageous alternative for the synthesis of glycosyl hydroxamates of biological interest. PMID- 17465569 TI - Gas-phase diastereoselectivity of secondary 5-substituted (X)-adamant-2-yl (X = F, Si(CH(3))(3)) cations. AB - Secondary 5-X-adamant-2-yl cations IX (X = F, Si(CH3)3) have been generated in the gas phase (total pressure = 760 Torr) from protonation-induced defluorination of epimeric 2-F-5-X-adamantanes 1X and their kinetic diastereoselectivity toward CH318OH investigated in the 40-160 degrees C range. The experimental results indicate that the facial selectivity of IX is insensitive to the composition of the starting 1X epimers as well as to the presence and the concentration of a powerful base (N(C2H5)3). This kinetic picture, supported by B3LYP/6-31G* calculations, is consistent with a single stable pyramidalized structure for IX, that is, (Z)-5-F-adamant-2-yl (I(Z)F) and (E)-5-Si(CH3)3-adamant-2-yl cations (I(E)Si). The temperature dependence of the IX diastereoselectivity lends support to the intermediacy of noncovalent adducts [IX*CH318OH], characterized by a specific C2-H+...O18(H)CH3 hydrogen bonding interaction. Their conversion to the covalently bonded O-methylated (Z)- (II(Z)X) and (E)-5-X-adamantan-2-ols (II(E)X; X = F, Si(CH3)3) is governed by activation parameters, whose magnitude depends on the specific IX face accommodating CH318OH. The gas-phase diastereoselectivity of IX toward CH318OH is compared to that exhibited in related gas-phase and solution processes. The emerging picture indicates that the factors determining the diastereoselectivity of IX toward simple nucleophiles in the gaseous and condensed media are completely different. PMID- 17465570 TI - Kinetic resolution in a bridgehead lithiation mediated by a chiral bis-lithium amide: assignment of the absolute configuration of clusianone. AB - A chiral base kinetic resolution of an advanced bicyclic intermediate enabled access to the polyprenylated phloroglucinol natural product (+)-clusianone in enantiomerically pure form. X-ray structure determination of another product obtained by the same method then allowed the absolute stereochemistry of (+) clusianone to be assigned. PMID- 17465571 TI - Toward an understanding of the acceleration of Diels-Alder reactions by a pseudo intramolecular process achieved by molecular recognition. A DFT study. AB - The pseudo-intramolecular Diels-Alder (DA) reaction between a 2-substituted furan (1) and a N-maleimide derivative (2) has been analyzed using DFT methods. Formation of two hydrogen bonds between the appendages on furan and maleimide derivatives favors thermodynamically the formation of a molecular complex (MC1) through an efficient molecular recognition process. The large enthalpy stabilization associated with the molecular recognition overcomes the unfavorable activation entropy associated with the bimolecular process. As a consequence, the subsequent DA reaction is clearly accelerated through a pseudo-intramolecular process. PMID- 17465572 TI - Approach to the homoerythrina alkaloids using a tandem N-alkylation/azomethine ylide cycloaddition. AB - Synthetic efforts toward the homoerythrina alkaloids 1-3 are described. Two separate model systems guided the pivotal [3 + 2] azomethine ylide cycloaddition cascade to form the A-C rings of these alkaloids. The cycloaddition precursors 63 and 68, prepared in nine and ten steps, respectively, from alkyne 47, each contain an enolizable ketone, a tethered electrophile, and an electron-poor dipolarophile. Heating 63 and 68 with the stannyl amine 17 generated demethoxyschelhammeridine 65 and demethoxyschelhammericine 70, the products of intramolecular azomethine ylide cycloadditions. Subsequent attempts to install the C-3 methoxy group of 1-3 are also described. PMID- 17465573 TI - A practical synthesis of a gamma-secretase inhibitor. AB - A practical and scaleable synthesis of the gamma-secretase inhibitor 1 is reported. The inhibitor consists of a central trisubstituted cyclohexane core with appended propionic acid, 2,5-difluorophenyl, and 4-chlorophenylsulfonyl moieties. Two alternative synthetic strategies, proceeding by way of a common disubstituted cyclohexanone derivative 5, were studied. In the preferred route, conjugate reduction of acrylonitrile derivative 4 with L-Selectride configures the desired relative stereochemistry of the cyclohexane core with >99.9:0.1 dr. A second strategy, based on catalyst-controlled hydrogenation of racemic cyclohexene derivative 2, is more convergent but less diastereoselective (up to 75:25 dr). The common cyclohexanone intermediate 5 was constructed by a regioselective Diels-Alder condensation of a 1,1-disubstituted vinyl sulfone 6 with 2-trimethylsiloxybutadiene. PMID- 17465574 TI - Extracellular carbohydrate metabolites from Streptomyces coelicolor A3(2). AB - A new ribose trisaccharide, alpha-Ribf-(1-->2)-alpha-Ribf-(1-->3)-alpha-Ribf (1), was isolated together with 5-O-(alpha-mannosyl)-myo-inositol (2), 2-O-(alpha mannosyl)-myo-inositol (3), trehalose (4), and d-ribulose (5) from a submerged cultivation of Streptomyces coelicolor A3(2). The structures of these compounds were elucidated by spectroscopic and chemical methods. Concentrations of these compounds in the medium were in the range from 0.04 (1) to 0.5 (4) mg/mL. PMID- 17465575 TI - Adsorption of designed pyrimidine derivative ligands on an activated carbon for the removal of Cu(II) ions from aqueous solution. AB - The adsorption of five Nalpha-substituted amino acids with a 5-nitroso-6-oxo pyrimidine as substituent on a commercial activated carbon (AC) has been studied in aqueous solution at several pH values. The adsorption processes of these organic compounds have been analyzed on the basis of the electrolytic behavior of the adsorbates. In all cases, the adsorption process is highly irreversible due to strong pi-pi interactions between the arene centers of the AC and the pyrimidine residue of the adsorbates. This interaction is consistent with XPS data and HOMO-LUMO theoretical calculations. The adsorption of these organic compounds provides a new route for the functionalization of the AC surface with carboxyl groups. In addition, the adsorption capacity of the AC/organic compound systems for Cu(II) ions in aqueous solution has been studied at different pH values. These systems show an increase of the adsorption capacity for Cu(II) compared to the AC, which is related to the AC functionalization with carboxyl groups due to the adsorbed organic compounds. PMID- 17465576 TI - Design of a superhydrophobic surface using woven structures. AB - The relationship between surface tension and roughness is reviewed. The Cassie Baxter model is restated in its original form, which better describes the most general cases of surface roughness. Using mechanical and chemical surface modification of nylon 6,6 woven fabric, an artificial superhydrophobic surface was prepared. A plain woven fabric mimicking the Lotus leaf was created by further grafting 1H,1H-perfluorooctylamine or octadecylamine to poly(acrylic acid) chains which had previously been grafted onto a nylon 6,6 woven fabric surface. Water contact angles as high as 168 degrees were achieved. Good agreement between the predictions based on the original Cassie-Baxter model and experiments was obtained. The version of the Cassie-Baxter model in current use could not be applied to this problem since the surface area fractions in this form is valid only when the liquid is in contact with a flat, porous surface. The angle at which a water droplet rolls off the surface has also been used to define a superhydrophobic surface. It is shown that the roll-off angle is highly dependent on droplet size. The roll-off angles of these superhydrophobic surfaces were less than 5 degrees when a 0.5 mL water droplet was applied. PMID- 17465577 TI - Evaporation of water droplets on polymer surfaces. AB - The evaporation of water droplets on polymer surfaces was investigated by using a digital image analysis technique. There were three distinct stages in the water evaporation process: a constant contact area mode, a constant contact angle mode, and a mixed mode that is independent of both the initial quantity of water droplets and the hydrophobic properties of the polymer surfaces. The physical factors influencing the first and second transitions in the evaporation process were found to be the attainment of the receding angle on the polymer surfaces and the Marangoni instability in the evaporating water droplets, which result from the concentration gradient of contaminants. This study also provides qualitative information about the microfluid flows inside the evaporating water droplets and the morphology of drying stains on polymer surfaces. The contaminants were found to be concentrated at the perimeter of the stains, in agreement with the observed outward microfluid flow in the mixed mode of the evaporation process. PMID- 17465578 TI - Control of the microdomain orientation in block copolymer thin films with homopolymers for lithographic application. AB - Block copolymer lithography is a promising method for fabricating periodical nanopatterns of less than 20 nm by self-assembly and can be applicable for fabricating patterned magnetic media with a recording density over 1 Tb/in.2. We found a simple technique to control the orientation of cylindrical microdomains in thin films. Simply by mixing polystyrene-block-poly(methyl methacrylate) (PS-b PMMA) diblock copolymers with the homopolymer (PS or PMMA) of the major component, we could align the cylindrical microdomains perpendicular to the film surface. The added homopolymer induces conformational entropic relaxation of the block chains in microdomain space and stabilizes the perpendicular orientation of hexagonally packed cylindrical microdomains. Thus formed perpendicular cylinders can be readily aligned in a regular array with a grating substrate. PMID- 17465579 TI - Macroscopic modeling of the surface tension of polymer-surfactant systems. AB - Polymer-surfactant mixtures are increasingly being used in a wide range of applications. Weakly interacting systems, such as SDS/PEO and SDS/PVP, comprise ionic surfactants and neutral polymers, while strongly interacting systems, such as SDS/POLYDMDAAC and C12TAB/NaPSS, comprise ionic surfactants and oppositely charged ionic polymers. The complex nature of interactions in the mixtures leads to interesting and surprising surface tension profiles as the concentrations of polymer and surfactant are varied. The purpose of our research has been to develop a model to explain these surface tension profiles and to understand how they relate to the formation of different complexes in the bulk solution. In this paper we show how an existing model based on the law of mass action can be extended to model the surface tension of weakly interacting systems, and we also extend it further to produce a model for the surface tension of strongly interacting systems. Applying the model to a variety of strongly interacting systems gives remarkable agreement with the experimental results. The model provides a sound theoretical basis for comparing and contrasting the behavior of different systems and greatly enhances our understanding of the features observed. PMID- 17465580 TI - Lateral diffusion coefficients of an eicosanyl-based bisglycerophosphocholine determined by PFG-NMR and FRAP. AB - We report the lateral diffusion properties of 2,2'-di-O-decyl-3,3'-di-O (eicosanyl)-bis-(rac-glycero)-1,1'-diphosphocholine (C20BAS) using pulsed-field gradient NMR (PFG-NMR) and fluorescence recovery after photobleaching (FRAP). C20BAS membranes display a melting transition at Tm = 15.7 degrees C, as determined by differential scanning calorimetry and 31P NMR chemical shift anisotropy. The lateral diffusion coefficient of C20BAS, as determined by PFG-NMR and FRAP, at 25 degrees C, were DPFG-NMR = 1.9 +/- 0.6 x 10(-8) cm2/s and DFRAP C20BAS = 1.2 +/- 0.1 x 10(-8) cm2/s, respectively. In comparison, the lateral diffusion coefficient of the monopolar phospholipid, 1-palmitoyl-2-oleoyl-sn glycero-3-phosphocholine (POPC), was 1.8 +/- 0.9 x 10(-8) and 2.5 +/- 0.9 x 10( 8) cm2/s using PFG-NMR and FRAP, respectively. PMID- 17465581 TI - Probing protein adsorption onto mercaptoundecanoic acid stabilized gold nanoparticles and surfaces by quartz crystal microbalance and zeta-potential measurements. AB - The adsorption characteristics of three proteins [bovine serum albumin (BSA), myoglobin (Mb), and cytochrome c (CytC)] onto self-assembled monolayers of mercaptoundecanoic acid (MUA) on both gold nanoparticles (AuNP) and gold surfaces (Au) are described. The combination of quartz crystal microbalance measurements with dissipation (QCM-D) and pH titrations of the zeta-potential provide information on layer structure, surface coverage, and potential. All three proteins formed adsorption layers consisting of an irreversibly adsorbed fraction and a reversibly adsorbed fraction. BSA showed the highest affinity for the MUA/Au, forming an irreversibly adsorbed rigid monolayer with a side-down orientation and packing close to that expected in the jamming limit. In addition, BSA showed a large change in the adsorbed mass due to reversibly bound protein. The data indicate that the irreversibly adsorbed fraction of CytC is a monolayer structure, whereas the irreversibly adsorbed Mb is present in form of a bilayer. The observation of stable BSA complexes on MUA/AuNPs at the isoelectric point by zeta-potential measurements demonstrates that BSA can sterically stabilize MUA/AuNP. On the other hand, MUA/AuNP coated with either Mb or CytC formed a reversible flocculated state at the isoelectric point. The colloidal stability differences may be correlated with weaker binding in the reversibly bound overlayer in the case of Mb and CytC as compared to BSA. PMID- 17465582 TI - Mechanism and modeling of nanorod formation from nanodots. AB - A population balance model based on Smoluchowski aggregation kinetics is developed to explain the formation of nanorods from a colloidal suspension of spherical nanoparticles (nanodots). Our model shows that linear pearl-chain aggregates form by the oriented attachment (OA) of nanodots during the early stages of synthesis, since it occurs with a time scale smaller than the coalescence time scale of nanodots present within an aggregate. The slower coalescence step leads to the transformation of the linear pearl-chain aggregate into a smooth nanorod over a longer time scale of many hours, as observed in experiments. The attachment kinetics is modeled by a modified Brownian collision frequency, with the latter decreasing with nanorod length, leading to the experimentally observed slower growth in nanorod length at longer times. The collision frequency also includes the effects of attractive dipole-dipole and van der Waals interactions between nanodots, which are primarily responsible for OA. Our model predictions are general, and they compare favorably with available experimental data in the literature on the distribution of the aspect ratio (length to diameter) of ZnO and ZnS nanorods over different time scales. PMID- 17465583 TI - Measurement of the charge number per adsorbed molecule and packing densities of self-assembled long-chain monolayers of thiols. AB - We have applied a recently developed method (Langmuir 2006, 22, 5509-5519) to determine charge numbers per adsorbed molecule and packing densities in self assembled monolayers (SAMs) of octadecanethiol (C18SH), a representative long chain thiol. Our method yields values of area per molecule that are physically reasonable, in contrast to the popular reductive desorption method, which gives molecular areas that are smaller than those determined by the van der Waals radii. In a nonadsorbing electrolyte, we were able to model the dependence of the charge number per adsorbed molecule on the electrode potential, taking into account that the desorption process is a substitution reaction between the solvent and the adsorbate. We have also shown that the charge number per adsorbed thiol is affected by the specific adsorption of the anion of the electrolyte. In the latter case, the thiol competes for adsorption sites at the surface not only with water but also with the anion of the electrolyte, and this competition has an effect on the measured charge number. PMID- 17465584 TI - Anisotropic wetting characteristics on submicrometer-scale periodic grooved surface. AB - Submicrometer-scale periodic structures consisting of parallel grooves were prepared on azobenzene-containing multiarm star polymer films by laser interference. The wetting characteristics on the patterned surfaces were studied by contact angle measurements. Macroscopic distortion of water drops was found on such small-scale surface structures, and the contact angles measured from the direction parallel to the grooves were larger than those measured from the perpendicular direction. A thermodynamic model was developed to calculate the change in the surface free energy as a function of the instantaneous contact angle when the three-phase contact line (TPCL) moves along the two orthogonal directions. It was found that the fluctuations, i.e., energy barriers, on the energy versus contact angle curves are crucial to the analysis of wetting anisotropy and contact angle hysteresis. The calculated advancing and receding contact angles from the energy versus contact angle curves were in good agreement with those measured experimentally. Furthermore, with the groove depth increasing, both the degree of wetting anisotropy and the contact angle hysteresis perpendicular to the grooves increased as a result of the increase in the energy barrier. The theoretical critical value of the groove depth, above which the anisotropic wetting appears, was determined to be 16 nm for the grooved surface with a wavelength of 396 nm. On the other hand, the effect of the groove wavelength on the contact angle hysteresis perpendicular to the grooves was also interpreted on the basis of the thermodynamic model. That is, with the wavelength decreasing, the contact angle hysteresis increased due to the increase in the number of energy barriers. These results may provide theoretical evidence for the design and application of anisotropic wetting surface. PMID- 17465585 TI - Spatially organized free-standing poly(p-xylylene) nanowires fabricated by vapor deposition. AB - Thin films of poly(chloro-p-xylylene) (PPXC) grown by the pyrolysis and evaporation of chloro-[2.2]paracyclophane in an evacuated chamber contain free standing, slanted, parallel columns that are 50 mum long and are assemblies of 50 to 100-nm-diameter nanowires, which thus can have an unprecedented aspect ratio as high as 1000:1. The nanostructured thin films organize spatially with a chemical structure similar to that of planar PPXC thin films, but the former also possess nanostructured morphology. Nanostructured thin films of poly(p-xylylene) (PPX) and its derivatives shall be useful as functionalized interfaces for antifouling coatings and biomedical devices. PMID- 17465586 TI - Elucidating the mechanism of cellular uptake and removal of protein-coated gold nanoparticles of different sizes and shapes. AB - We investigated the mechanism by which transferrin-coated gold nanoparticles (Au NP) of different sizes and shapes entered mammalian cells. We determined that transferrin-coated Au NP entered the cells via clathrin-mediated endocytosis pathway. The NPs exocytosed out of the cells in a linear relationship to size. This was different than the relationship between uptake and size. Furthermore, we developed a mathematical equation to predict the relationship of size versus exocytosis for different cell lines. These studies will provide guidelines for developing NPs for imaging and drug delivery applications, which will require "controlling" NP accumulation rate. These studies will also have implications in determining nanotoxicity. PMID- 17465587 TI - Proceedings of the 2006 World Congress on the Science and Medicine of the Marathon. PMID- 17465588 TI - An overview of the 1976 New York academy of science meeting. AB - In the late 1960s and early 1970s, endurance running was gaining popularity at an exponential rate. Paul Milvy organised the scientific and medical conference entitled 'The Marathon: Physiological, Medical, Epidemiological, and Psychological Studies' associated with 1976 New York Marathon and sponsored by the New York Academy of Sciences to provide a forum for scientists to interact and to publish the proceedings to serve as a resource for future sports medicine research. The conference was a major success and the published proceedings have continued to be a significant resource for those in exercise science and sports medicine over the past 30 years. The challenge for this meeting is to bring forward the many new findings on marathon running and fill many of the gaps in our knowledge of exercise performance and human health that have occurred since 1976. PMID- 17465589 TI - The history of the marathon : 1976-present. AB - Since the 1976 New York City Marathon, the first 'urban tour' marathon, the sport has exploded on a global basis. Once limited largely to the Olympics, the Boston Marathon and several others, marathons are now held worldwide, with dozens of major cities holding races that attract upwards of 15,000 participants. The growth of women's and masters' age group (>40 years old) marathon racing has been particularly startling. PMID- 17465590 TI - The evolution of marathon running : capabilities in humans. AB - Humans have exceptional capabilities to run long distances in hot, arid conditions. These abilities, unique among primates and rare among mammals, derive from a suite of specialised features that permit running humans to store and release energy effectively in the lower limb, help keep the body's center of mass stable and overcome the thermoregulatory challenges of long distance running. Human endurance running performance capabilities compare favourably with those of other mammals and probably emerged sometime around 2 million years ago in order to help meat-eating hominids compete with other carnivores. PMID- 17465591 TI - Marathon medical support historical perspectives : 'from cradle to averting the grave'. AB - The history of marathon medical support is largely undocumented and anecdotal. Modern mass-participation marathon events attract thousands of variably trained and acclimatised participants to an event that is physically very demanding, in some cases covers difficult terrain, and may be held under stressful climatic conditions. Medical support for mass-participation events such as the marathon is directed at minimising the potential risks and avoiding the worst scenario where local medical emergency services and hospitals are flooded with casualties. PMID- 17465592 TI - American women in the marathon. AB - American women have made great advances in the sport of marathon running over the past 4 decades. The purpose of this study was to examine the trend of marathon times among American female runners between 1976 and 2005, and to compare physiological characteristics of male and female runners. The best marathon times of American female and male marathon runners for each year (1976-2005) were collected from several published sources. Two research studies were reviewed that examined a variety of physiological variables of female and male elite distance runners. While the best marathon times of American men have remained fairly constant in recent decades ( approximately 2:10:00), the best times of American women have decreased dramatically from 2:47:10 in 1976 to 2:21:25 in 2005, a decrease of 15.6% over the 30-year period. The physiological characteristics of elite American female marathon runners differ from those of elite male marathon runners (e.g. maximal oxygen uptake = 67.1 +/- 4.2 mL/kg/min vs 74.1 +/- 2.6 mL/kg/min). These differences are comparable with the differences seen in marathon performance. Over the past 30 years, participation by women in marathon running has grown dramatically and during that same period the marathon performances of women have improved at a remarkable rate. PMID- 17465593 TI - Can children and adolescents run marathons? AB - There are few data on youth marathon runners and the most commonly asked questions of "can children run marathons?" and "what are the health consequences?" remain unanswered. Expert opinion is split with regard to running this distance at a young age. There have been many thousands of finishers <18 years old at the Los Angeles Marathon in an organised programme for youth running and nearly 300 finishers in at the Twin Cities Marathon. The youngest child in these data sets was 7 years old. There has not been any significant medical injury at these events. Children who choose of their own accord to participate in marathon training should be allowed to do so as long as their social, academic, psychological and physiological development is not disrupted. Follow-up studies of these young runners would help evaluate the long-term health consequences of long distance running and shape future recommendations. PMID- 17465594 TI - Marathon runners: how do they age? AB - Marathon running performance among men and women is generally fastest, as indicated by world record performances, when individuals are 25-35 years old. The time to complete a marathon gradually increases with age, with substantial losses in performance after the age of 70 years. A decline in cardiovascular capacity of 0.5% per decade occurs in highly trained distance runners, while a 1.0% and 1.5% decline per decade occurs in moderately trained and untrained individuals, respectively. In middle-aged veteran runners, skeletal muscle continues to have high aerobic potential, while a decline in muscle cell size and contractile performance are apparent. These changes in the skeletal muscle profile may contribute to distance running performance with age. The changes in physiological function and running performance with age are closely related to the level of distance run training. Current research supports the concept that continued running late into life attenuates a decline in physiological function with age and is beneficial for overall health. PMID- 17465595 TI - Physiological regulation of marathon performance. AB - Running a marathon at the fastest speed possible appears to be regulated by the rate of aerobic metabolism (i.e. marathon oxygen uptake) of a limited amount of carbohydrate energy (i.e. muscle glycogen and blood glucose) and the velocity that can be maintained without developing hyperthermia. According to a model proposed by Joyner in 1991, people possess the physiological ability to run a marathon in approximately 1:58:00. This could be accomplished if the current world record pace for the 'half-marathon' is maintained for the entire marathon. The ultimate limit to marathon performance might be dictated by the limits of running economy and a recruitment of the running musculature with a pattern that minimises fatigue, possibly by spreading the work over many motor neuron. PMID- 17465596 TI - Thermoregulatory function during the marathon. AB - Marathon races are performed over a broad range of environmental conditions. Hyperthermia is a primary challenge for runners in temperate and warm weather, but hypothermia can be a concern during cool-wet or cold conditions. Body temperature during the marathon is a balance between metabolic heat production and exchange with the environment described by the heat balance equation. During exercise, core temperature is proportional to the metabolic rate and largely independent of a wide range of environmental conditions. In temperate or cool conditions, a large skin-to-ambient temperature gradient facilitates radiant and convective heat loss, and reduces skin blood flow requirements, which may explain the tolerance for high core temperature observed during marathons in cool conditions. However, in warmer environments, skin temperatures and sweating rates increase. In addition, greater skin blood flow is required for heat loss, magnifying thermoregulatory and circulatory strain. The combined challenge of exercise and environment associated with marathon running can substantially challenge the human thermoregulatory system. PMID- 17465597 TI - Running economy : the forgotten factor in elite performance. AB - Running performance depends on maximal oxygen uptake (VO(2max)), the ability to sustain a high percentage of VO(2max) for an extended period of time and running economy. Running economy has been studied relatively less than the other factors. Running economy, measured as steady state oxygen uptake (VO(2)) at intensities below the ventilatory threshold is the standard method. Extrapolation to a common running speed (268 m/min) or as the VO(2) required to run a kilometer is the standard method of assessment. Individuals of East African origin may be systematically more economical, although a smaller body size and a thinner lower leg may be the primary factors. Strategies for improving running economy remain to be developed, although it appears that high intensity running may be a common element acting to improve economy. PMID- 17465598 TI - Marathon performance in thermally stressing conditions. AB - It is generally appreciated that warm weather negatively affects marathon running performance. This brief review summarises the historical literature on this topic and recent work that our laboratory has performed to quantify the impact of weather on marathon running performance. Using 140 race-years of data, we have demonstrated that marathon performance times slow progressively as weather warms above 5-10 degrees C wet bulb globe temperature, that men and women are affected similarly, but slower runners suffer a greater performance penalty than elite runners. The recent generation of a nomogram that predicts changes in finishing time consequent to changes in weather conditions offers runners and coaches a tool for use in developing marathon race strategy. PMID- 17465599 TI - Strategies for optimising marathon performance in the heat. AB - During the next few years, several top-level sport competitions involving endurance running, such as the marathon, will occur in hot/humid Asian summertime weather. These competitions include the World Athletics Championships, the World Student Games and the Summer Olympic Games. Well prepared athletes have a fitness level so high that they can produce more metabolic heat than can be dissipated, and thus run the risk of encountering heat illness unless they fine-tune their pace, making it compatible with manageable heat production. The ideal starting time for such long races would be early evening, so that athletes encounter steadily improving weather (less sun, gradual cooling and minimum rise in humidity). However, current plans for competition scheduling have the marathon races starting in the early morning hours. Thus, as the race develops and athletes experience a trend towards dehydration and energy depletion, weather conditions will likely deteriorate and impact negatively upon performance. Suggestions are provided for optimising performance under these circumstances. PMID- 17465600 TI - Metabolic adaptations to marathon training and racing. AB - During the past 30 years, considerable progress has been made in our understanding of the cellular and molecular factors regulating fuel metabolism during exercise. In particular, advancements in the fields of exercise biochemistry and cell signalling have helped elucidate the mechanism(s) by which perturbations in energy status are monitored inside contracting muscle cells, and have helped identify target molecules that increase fuel supply to maintain adenosine triphosphate concentration. In this brief commentary, we summarise some of the major cellular and molecular adaptations in human skeletal muscle resulting from the intense endurance training required to run a marathon. PMID- 17465601 TI - Regulation of substrate use during the marathon. AB - The energy required to run a marathon is mainly provided through oxidative phosphorylation in the mitochondria of the active muscles. Small amounts of energy from substrate phosphorylation are also required during transitions and short periods when running speed is increased. The three inputs for adenosine triphosphate production in the mitochondria include oxygen, free adenosine diphosphate and inorganic phosphate, and reducing equivalents. The reducing equivalents are derived from the metabolism of fat and carbohydrate (CHO), which are mobilised from intramuscular stores and also delivered from adipose tissue and liver, respectively. The metabolism of fat and CHO is tightly controlled at several regulatory sites during marathon running. Slower, recreational runners run at 60-65% maximal oxygen uptake (VO(2max)) for approximately 3:45:00 and faster athletes run at 70-75% for approximately 2:45:00. Both groups rely heavily on fat and CHO fuels. However, elite athletes run marathons at speeds requiring between 80% and 90% VO(2max), and finish in times between 2:05:00 and 2:20:00. They are highly adapted to oxidise fat and must do so during training. However, they compete at such high running speeds, that CHO oxidation (also highly adapted) may be the exclusive source of energy while racing. Further work with elite athletes is needed to examine this possibility. PMID- 17465602 TI - Protein metabolism and endurance exercise. AB - Prolonged endurance exercise stimulates whole-body protein turnover (synthesis and degradation) but it remains contentious whether this translates into an increased net protein oxidation or dietary requirement for protein. Skeletal muscle is the major energy consumer during exercise and the oxidation of branched chain amino acids (BCAA) is increased several-fold, suggesting an increased requirement for fuel. Increased BCAA oxidation has been proposed to impair aerobic energy provision during prolonged exercise, but there is little evidence to support this theory. Endurance training blunts the acute exercise-induced increase in whole-body protein turnover and skeletal BCAA oxidation at a given work intensity. However, training also increases the maximal capacity for skeletal muscle BCAA oxidation, as evidenced by a higher maximal activity of the rate-determining enzyme branched-chain oxo acid dehydrogenase. Exercise-induced changes in protein metabolism are affected by nutritional status, with high carbohydrate availability (as typically practiced by endurance athletes) generally associated with reduced net protein utilisation. Ingestion of protein with carbohydrate improves net protein balance during exercise and recovery compared with carbohydrate alone, but it remains to be determined whether this practice facilitates the adaptive response to chronic training. PMID- 17465603 TI - Lactate: link between glycolytic and oxidative metabolism. AB - Once thought to be the consequence of oxygen lack in contracting skeletal muscle, the glycolytic product lactate is formed and utilised continuously under fully aerobic conditions. 'Cell-cell' and 'intracellular lactate shuttle' concepts describe the roles of lactate in delivery of oxidative and gluconeogenic substrates as well as in cell signalling. Examples of cell-cell shuttles include lactate exchanges (i) between white-glycolytic and red-oxidative fibres within a working muscle bed; (ii) between working skeletal muscle and heart; and (iii) between tissues of net lactate release and gluconeogenesis. Lactate shuttles exist in diverse tissues including in the brain, where a shuttle between astrocytes and neurons is linked to glutamatergic signalling. Because lactate, the product of glycogenolysis and glycolysis, is disposed of by oxidative metabolism, lactate shuttling unites the two major processes of cellular energy transduction. Lactate disposal is mainly through oxidation, especially during exercise when oxidation accounts for 70-75% of removal and gluconeogenesis the remainder. Lactate flux occurs down proton and concentration gradients that are established by the mitochondrial lactate oxidation complex. Marathon running is a power activity requiring high glycolytic and oxidative fluxes; such activities require lactate shuttling. Knowledge of the lactate shuttle is yet to be imparted to the sport. PMID- 17465604 TI - Nutrition strategies for the marathon : fuel for training and racing. AB - Muscle glycogen provides a key fuel for training and racing a marathon. Carbohydrate 'loading' can enhance marathon performance by allowing the competitor to run at their optimal pace for a longer period before fatiguing. For the well trained runner, this may be achieved by tapering exercise over the final days before the marathon and ensuring carbohydrate intakes of 10-12 g/kg/day over the 36-48 hours prior to the race. Sports nutrition guidelines recommend that the runner consumes sufficient carbohydrate to promote restoration of muscle glycogen between training sessions. This strategy should allow the runner to 'train harder' and recover optimally between workouts. A recent hypothesis suggests that runners might 'train smarter' by training with low glycogen stores, since this might promote greater stimulation of the training response. However, there is no evidence that a low carbohydrate diet enhances the outcomes of training or provides benefits as a depletion phase prior to carbohydrate loading. In fact, a low carbohydrate diet may even impair performance if carried out for extended periods. If there are benefits to manipulating glycogen stores for some workouts, this is likely to happen as the natural outcome of the periodisation of the high volume programme of an elite runner. PMID- 17465605 TI - Low energy availability in the marathon and other endurance sports. AB - Energy availability is the amount of dietary energy remaining after exercise training for all other metabolic processes. Excessively low energy availability impairs reproductive and skeletal health, although genetics and age may alter an individual's initial conditions and sensitivity when low energy availability is imposed. Many marathon runners and other endurance athletes reduce energy availability either (i) intentionally to modify body size and composition for improving performance; (ii) compulsively in a psychopathological pattern of disordered eating; or (iii) inadvertently because there is no strong biological drive to match energy intake to activity-induced energy expenditure. Inadvertent low energy availability is more extreme when consuming a low fat, high carbohydrate diet. Low energy availability, reproductive disorders, low bone mineral density and stress fractures are more common in female than male athletes. Functional menstrual disorders caused by low energy availability should be diagnosed by excluding diseases that also disrupt menstrual cycles. To determine energy availability (in units of kilocalories or kilojoules per kilogram of fat-free mass), athletes can record their diets and use diet analysis software to calculate energy intake, measure energy expenditure during exercise using a heart monitor and measure fat-free mass using a bioelectrical impedance body composition scale. All are commercially available at consumer prices. PMID- 17465606 TI - Fluid replacement and performance during the marathon. AB - The primary purpose of this review is to relate a universal strategy for replacing fluids to optimise marathon performance. A secondary purpose is to examine common 'matters of debate' that may modify fluid needs to include the importance of realistic convective air flow, metabolic water production and waters of association with glycogen. The metabolic demands of marathon running can result in substantial sweat losses and levels of dehydration consistent with compromised endurance performance. Recommendations are provided to individualise fluid intakes with the goal of preventing excessive dehydration (>2% body mass) as well as weight gain. The minor importance of 'matters of debate' to fluid replacement is also discussed. PMID- 17465607 TI - The role of salt and glucose replacement drinks in the marathon. AB - There is a large and growing body of scientific evidence that documents the benefits of ingesting salt and glucose (carbohydrates) during prolonged exercise. Those benefits include maintenance of cardiovascular function, enhanced carbohydrate oxidation, blunted decline in plasma sodium concentration and improved performance. The consumption of approximately 1g of carbohydrate per kilogram of bodyweight per hour appears sufficient to improve performance in prolonged exercise. Research also indicates that approximately 450mg of sodium per hour is the minimum amount required to maintain plasma volume and slow the decline in plasma sodium concentration that can accompany prolonged exercise in some runners. Adequate carbohydrate and electrolyte intake can be achieved by consuming a well formulated sports drink at regular intervals during exercise, in volumes designed to minimise dehydration. For marathon runners, this could range from approximately 400mL to >1.5L per hour, depending upon individual sweating rates. PMID- 17465608 TI - Exertional rhabdomyolysis and acute renal failure in marathon runners. AB - Strenuous exercise, including marathon running, can result in damage to skeletal muscle cells, a process known as exertional rhabdomyolysis. In most cases, this damage is resolved without consequence. However, when the damage is profound, there is a release of muscle proteins into the blood; one of these proteins, myoglobin, in high concentrations and under certain conditions (such as dehydration and heat stress) can precipitate in the kidneys, thereby resulting in acute renal failure. Although the marathon is a gruelling physiological challenge, with races sometimes run in hot and humid weather, acute renal failure is relatively infrequent. From case reports, a high proportion of marathon runners who developed acute renal failure had taken analgesics, had a viral or bacterial infection, or a pre-existing condition. The rare cases of acute renal failure in marathon runners may be a situation of the 'perfect storm' where there are several factors (heat stress, dehydration, latent myopathy, non-steroidal anti-inflammatory or other drug/analgesic use, and viral/bacterial infection) that, in some combination, come together to result in acute renal failure. PMID- 17465609 TI - Muscle cramping in the marathon : aetiology and risk factors. AB - Skeletal muscle cramps are commonly encountered in marathon runners by medical staff. However, the aetiology, and therefore management, of this condition is not well understood. Exercise-associated muscle cramping (EAMC) is defined as an involuntary, painful contraction of skeletal muscle during or immediately after exercise. In early anecdotal reports, cramps were associated with profuse sweating, together with changes in serum electrolyte concentrations. No mechanism explains how such imbalances in serum electrolytes result in localised muscle cramping. The 'muscle fatigue' hypothesis suggests that EAMC is the result of an abnormality of neuromuscular control at the spinal level in response to fatiguing exercise and is based on evidence from epidemiological studies, animal experimental data on spinal reflex activity during fatigue and electromyogram data recorded during bouts of acute cramping after fatiguing exercise. The development of premature muscle fatigue appears to explain the onset of EAMC. PMID- 17465610 TI - The role of sodium in 'heat cramping'. AB - 'Heat cramping' is defined here as severe, spreading, sustained, sharply painful muscle contractions that can sideline athletes. Not all cramps are alike, but three lines of evidence suggest heat cramping is caused by 'salty sweating', specifically by the triad of salt loss, fluid loss and muscle fatigue. The first line of evidence is historical. Dating back 100 years, heat cramping in industrial workers was alleviated by saline, and in a self-experiment, salt depletion provoked muscle cramping. The second line of evidence is from field studies of athletes. In tennis and football alike, heat-crampers tend to be salty sweaters. Some evidence also suggests that triathletes who cramp may lose more salt during the race than peers who do not cramp. The third line of evidence is practical experience with therapy and prevention. Intravenous saline can reverse heat cramping, and more salt in the diet and in sports drinks can help prevent heat cramping. For heat cramping, the solution is saline. PMID- 17465611 TI - Hyperthermia impairs brain, heart and muscle function in exercising humans. AB - Marathon running poses a severe challenge to multiple regulatory systems and cellular homeostasis, especially when performed in hot environments without fluid replacement. During exercise in the heat, the ensuing dehydration causes hyperthermia and the synergistic effects of both stressors reduce cardiac output and blood flow to muscle, skin, brain and possibly splanchnic tissues. The drop in blood flow beyond the regulatory adjustment to concurrent increases in blood oxygen content leads to reductions in oxygen delivery, suppressed muscle aerobic energy turnover and greater reliance of the exercising muscles on anaerobic metabolism before fatigue. The accelerated hyperthermia-mediated fatigue during prolonged and maximal exercise is preceded by functional alterations in multiple bodily systems including the brain, heart and muscle. It is proposed that the impaired marathon running performance in warm environments is associated with a greater thermal, cardiovascular and metabolic strain, and perception of effort that prevents marathon runners from running at their personal record speed without inducing an accelerated regulatory dysfunction in multiple bodily systems. PMID- 17465612 TI - The central governor model of exercise regulation applied to the marathon. AB - Two popular models hold that performance during exercise is limited by chemical factors acting either in the exercising muscles or in the brain producing either 'peripheral' or 'central' fatigue, respectively. A common feature of both models is that neither allows humans to 'anticipate' what will happen in the future and modify their exercise response accordingly. The peripheral fatigue model predicts that exercise terminates only after there has been catastrophic failure in one or more body systems and only when all the available motor units in the active muscles have been activated. The marathon race provides evidence that human athletes race 'in anticipation' by setting a variable pace at the start, dependent in part on the environmental conditions and the expected difficulty of the course, with the capacity to increase that pace near the finish. Marathoners also finish such races without evidence for a catastrophic failure of homeostasis characterised by the development of a state of absolute fatigue in which all the available motor units in their active muscles are recruited. These findings are best explained by the action of a central (brain) neural control that regulates performance in the marathon 'in anticipation' specifically to prevent biological harm. PMID- 17465613 TI - Heat exhaustion and dehydration as causes of marathon collapse. AB - This article reviews causes of marathon collapse related to physical exhaustion, heat exhaustion and dehydration. During severe exercise-heat stress (high skin and core temperatures), cardiac output can decrease below levels observed during exercise in temperate conditions. This reduced cardiac output and vasodilated skin and muscle can make it difficult to sustain blood pressure and perhaps cerebral blood flow. Dehydration can accentuate this cardiovascular strain. In contrast, excessive heat loss to the environment during cold weather may result in hypothermic collapse. Other factors contributing to post-race collapse might include reduced skeletal muscle pump activity and dehydration and prior heat stress mediated changes in cerebrovascular responses to orthostatic challenges. PMID- 17465615 TI - History and prevalence of doping in the marathon. AB - Anti-doping activism represents the mainstream of the current anti-doping campaign and includes both an ethical critique of doping and a call for the sharpening of sanctions. At the same time, evolving evidence indicates that systemic doping in some sports has existed on a larger scale than most observers thought possible. Doping has historical roots in the physiological demands of extreme endurance sport. The Victorian philosophy of sportsmanship and fair competition that emphasises the importance of a capacity for self-restraint and expresses a code of honour, once associated with the gentleman ideal, is the basis of the current anti-doping movement. The problem with this focus is the assumption that there is a contradiction between high-performance endurance sports and doping practices that is not found in the historical record. Compared with the hundreds of professional cyclists who have tested positive for banned substances in recent decades, the list of offenders in the marathon is very short. At the same time, the low number of positive tests is an inadequate basis for estimating the extent of doping among elite marathon runners today. A better understanding of this phenomenon would require something like anthropological fieldwork conducted in this subculture. PMID- 17465614 TI - Reduced peripheral resistance and other factors in marathon collapse. AB - Athletes sometimes collapse either during or after exercise. Conditions that cause collapse during exercise may be life-threatening and account for 10-15% of all exercise-related medical encounters at endurance sporting events. These conditions are not always 'sports-specific' and are managed according to the usual clinical care guidelines. The majority (>85%) of exercise-related collapse occurs after athletes have completed the event and key features are that patients are fully conscious and they have no clinical evidence for other serious medical conditions, but they have postural hypotension (blood pressure usually <100mm Hg) with unexpectedly low heart rates (<100 beats per minute). The cause of the postural hypotension appears to be the combination of physiological changes induced by exercise that tend to maintain a state of abnormally low peripheral vascular resistance for some hours after exercise. The most appropriate treatment for these patients is the supine position with the legs and pelvis above the level of the heart. Additional therapeutic interventions should be reserved for those who fail to respond adequately (blood pressure >100mm Hg; normal heart rate) to this simple treatment. PMID- 17465616 TI - Blood doping : infusions, erythropoietin and artificial blood. AB - As science marches on, athletes and coaches march close behind. Researchers have long been interested in how red cell mass and blood volume affect exercise capacity. Interest in blood doping soared after the 1968 Mexico City Olympics. Studies in the 1970s and 1980s suggested that transfusing red cells could speed endurance performance. Diverse athletes of the time were accused of blood doping. In the late 1980s, recombinant human erythropoietin (EPO) began to supplant transfusion for doping. EPO use is a suspect in nearly 20 deaths in 4 years in European cyclists. In the 1998 Tour de France, a team was ejected for using EPO and six other teams quit the race. The beat goes on; in recent years, diverse endurance and sprint athletes have been caught or accused of using EPO. Tests to detect EPO are improving but are not yet foolproof. As EPO tests improve, blood transfusion is back in vogue and some athletes may have infused artificial blood. Tests for detecting artificial blood also exist, but it seems it will take widespread, year-round, unannounced, out-of-competition testing and stern penalties to deter blood doping. PMID- 17465617 TI - Altitude training for the marathon. AB - For nearly 40 years, scientists and elite endurance athletes have been investigating the use of altitude in an effort to enhance exercise performance. While the results of many early studies on the use of altitude training for sea level performance enhancement have produced equivocal results, newer studies using the 'live high, train low' altitude training model have demonstrated significant improvements in red cell mass, maximal oxygen uptake, oxygen uptake at ventilatory threshold, and 3000m and 5000m race time. For the marathoner looking to add altitude training to their peak performance plans, residence at an altitude of 2000-2500m, a minimum of 20 hours per day, for 4 weeks, appears to hold the greatest potential for performance enhancement. Based on published mathematical models of marathon performance, a marathoner with a typical or average running economy who performed 'live high, train low' altitude training could experience an improvement of nearly 8.5 minutes (or approximately 5%) over the 26.2-mile race distance. PMID- 17465618 TI - Heat and cold : what does the environment do to the marathon runner? AB - The marathon poses a considerable physical challenge for athletes of all levels. When combined with high heat and humidity, not only is performance potentially compromised, but health and well-being are also at risk. There are well recognised effects of heat and hydration status on the cardiovascular and thermoregulatory systems that can account for the decreased performance and increased sensation of effort that are experienced when competing in the heat. Elevated exercise heart rate and core temperature at the same absolute exercise intensity are commonly reported. Dehydration occurring during exercise in the heat and results in reductions in stroke volume, cardiac output and blood pressure, as well as a marked decline in blood flow to the working muscles. Recent work suggests that hyperthermia may have a direct affect on the CNS and the brain may contribute to fatigue during prolonged exercise in a warm environment. At present, evidence supports a significant role of catecholaminergic neurotransmission, but there are a number of metabolic and circulatory perturbations occurring within the brain that may also be important in the fatigue process. PMID- 17465619 TI - Heat and cold: what does the environment do to marathon injury? AB - The medical work load seems to increase both with heat and humidity, and with cold and rainy conditions. Heat tolerance during exercise is variable and heat intolerance may contribute to collapse and increase medical encounters. Exposure to cold, wet conditions results in increasing incidence of hypothermia in exhausted marathon runners. Finish-line encounters and course dropouts increase as conditions cool and warm away from the most advantageous conditions in the 4.4 15 degrees C (40-59 degrees F) wet bulb globe temperature (WBGT) range. The risk of requiring medical attention and not finishing rises considerably when the WBGT is >15.5 degrees C (60 degrees F). Comparing the correlation coefficients of the Boston Marathon and Twin Cities Marathon data suggests that the risks of medical problems and not finishing are associated with the warmest temperature of the race and not the start temperature. The community consequences of races conducted in hot and humid conditions can be significant, particularly when the WBGT is >15.5 degrees C. The emergency medical systems can be overwhelmed with a surge of patients, some very ill, and the emergency call response times drop to unacceptable levels blocking access for the citizens of the community. With respect to marathon encounters, heat stress increases both the finish-line medical encounter rate and the on course drop-out rate, and seems to increase the incidence of hyponatraemia and heat stroke. Cold conditions increase the drop-out rate along the course and, if associated with wet conditions, also increase the encounter rate. PMID- 17465620 TI - The psychology of the marathoner : of one mind and many. AB - The unique physiological attributes of marathoners have long been recognised, but until the pioneering research of Morgan and Pollock (1977) little was known about their psychological characteristics. Their work revealed marathoners have significantly better mental health compared with non-athletes, with desirable mental health variables being most pronounced in elite competitors. It was also found that during competition, elite marathoners typically utilise a unique cognitive strategy labelled 'association', whereby they regulate pace based upon bodily sensations including pain and effort. More recent research indicates there are considerable individual differences in the psychological responses of marathoners to the stressors associated with training and competition, and in some cases negative emotions traditionally presumed to be harmful actually benefit performance. This brief review will highlight findings of psychological research involving marathoners and other endurance athletes, distinguishing between characteristics common among groups (i.e. nomothetic) with those particular to individuals (i.e. ideographic) or sub-groups of elite and non-elite competitors. PMID- 17465621 TI - Monitoring and titrating symptoms : a science-based approach to using your brain to optimise marathon running performance. AB - A key challenge to optimising marathon running performance is to train with adequate frequency, duration and intensity as well as get enough recovery to optimise biological adaptations underlying performance. Some marathon runners train inadequately and underperform, while others perform poorly because they become injured or develop staleness in response to overtraining. Staleness, a depression-like syndrome, could plausibly be caused by overtraining-induced molecular and cellular changes in brain circuits involved in depression or other related mood states such as anger, fatigue, vigor and confusion. The central thesis of this paper is that easily assessed resting and/or exercise symptoms, valid markers of either difficult-to-access, expensive-to-assess or unmeasurable brain circuits, can be used to optimise marathon running performance by helping to avoid either inadequate training or excessive training resulting in staleness. Available models of human performance and relevant data, admittedly incomplete at the present time, suggest that marathon runners may benefit from systematically using symptom responses to training in order to aid in adjusting training loads for the purpose of optimising training. As this approach is better linked with neuroscience and neuroimaging findings, it could be refined and prove to be useful for elite as well as non-elite marathon runners. PMID- 17465622 TI - Marathon training and immune function. AB - Many components of the immune system exhibit adverse change after marathon-type exertion. These immune changes occur in several compartments of the immune system and body (e.g. the skin, upper respiratory tract mucosal tissue, lung, peritoneal cavity, blood and muscle). Of all immune cells, natural killer (NK) cells, neutrophils and macrophages (of the innate immune system) exhibit the greatest changes in response to marathon competition, both in terms of numbers and function. Many mechanisms appear to be involved, including exercise-induced changes in stress hormone and cytokine concentrations, body temperature changes, increases in blood flow and dehydration. During this 'open window' of immune dysfunction (which may last between 3 and 72 hours, depending on the immune measure), viruses and bacteria may gain a foothold, increasing the risk of subclinical and clinical infection. Of the various nutritional and pharmacological countermeasures to marathon-induced immune perturbations that have been evaluated thus far, ingestion of carbohydrate beverages during intense and prolonged exercise has emerged as the most effective. However, carbohydrate ingestion during a marathon attenuates increases in plasma cytokines and stress hormones, but is largely ineffective against changes in other immune components including suppression of NK and T-cell function, and salivary IgA output. Other countermeasures, such as glutamine, antioxidant supplements and ibuprofen, have had disappointing results and thus the search for companion agents to carbohydrate continues. PMID- 17465623 TI - Strategies to enhance immune function for marathon runners : what can be done? AB - Marathoners are at an increased risk of developing upper respiratory tract infections (URTIs) following races and periods of hard training, which are associated with temporary changes in the immune system. The majority of the reported changes are decreases in function or concentration of certain immune cells. During this period of immune suppression, by some referred to as an 'open window' in immune function, it has been hypothesised that viruses and bacteria might gain a foothold, which would increase the risk of infections. In light of this, nutritional interventions that can enhance immune function and reduce the risk of URTIs have been sought. This paper focuses on the effect of glutamine, vitamin C, bovine colostrum and glucose. Although, some of these supplements can affect the physiological and immune changes associated with marathon racing, none of the supplements discussed have consistently been shown to reduce the risk of URTIs and therefore cannot be recommended for use as enhancers of immune function in marathon runners. PMID- 17465624 TI - Biomechanical factors contributing to marathon race success. AB - During marathon running, the body's metabolic resources become depleted and musculoskeletal stress and fatigue begin to hinder performance, making efficient running a must. Biomechanical studies of long distance running have tried to identify how body structure and running mechanics interact with performance, economy and injury, and typically have involved groups of subjects. While moderate relationships have been identified, the outcomes include conflicting results, vague conclusions and unclear consequences. Easily identifiable and universally applicable patterns of efficient movement have not been found. An alternative avenue of research is described that concentrates on identifying how an individual runner's structure and functional abilities influence performance, economy and injury. It is hoped that when such an approach identifies important relationships for individuals, the patterns identified will lead to a more general understanding of the underlying mechanisms. PMID- 17465625 TI - Genotypes and distance running : clues from Africa. AB - A look at the medal podium in almost any international sporting competition reveals that some athletes and certain countries enjoy regular success in particular events. While environmental influences such as training and diet are important, it is likely that there is also some genetic component to elite athletic performance. One of the most compelling examples of athletic domination is that of east African runners in international distance running competition. This phenomenon has led to the suggestion that east Africans possess some inherent genetic advantage predisposing them to superior athletic performances. The concurrent success of athletes of west African ancestry in sprint events also appears to have augmented this belief given their similar skin colour. A growing body of evidence suggests that genetic variation does influence athletic performance, yet despite the speculation that African athletes have a genetic advantage for physical performance, there is no genetic evidence to suggest that this is the case. The only available genetic studies of elite African athletes do not find that these athletes possess a unique genetic makeup; rather, they serve to highlight the high degree of genetic diversity in east African populations and also among elite east African athletes. PMID- 17465626 TI - Marathon race medical administration. AB - Marathon race medical coverage involves coordinating many organisations, personnel and government agencies. An infrastructure of aid stations, finish-line medical tents, ambulance services, communication networks and transport vehicles support the medical professionals covering the event. Knowledge of the volume and type of medical problems that arise during a marathon allow the medical director to assemble the necessary components for a safe event. Published data and/or historical race information will assist in planning for anticipated medical utilisation. Organisers must prepare for uncommon but possible catastrophic medical events such as cardiac arrest and exertional heat stroke. Advances in point-of-care medical devices and significant increases in training and education of medical professionals providing care at these events have greatly improved the quality of on-site medical care at marathon races. PMID- 17465627 TI - Exercise-associated collapse care matrix in the marathon. AB - Exercise-associated collapse (EAC) was developed as a marathon collapse classification matrix to speed clinical decision making and improve care. The definition was simply stated as the need for assistance during or immediately after endurance activity that specifically excluded cardiac arrest, insulin shock, anaphylaxis, trauma, skin conditions and orthopedic injuries. The presenting symptoms were neither specific nor sensitive and the initial classification, based on presenting rectal temperature, neurological status and ability to walk, was intended to shape the on-site medical intervention. The treatment of EAC centres on fluid redistribution and replacement to improve cerebral and core organ circulation and body temperature correction if needed. Most runners are discharged from the medical area in the company of another person. PMID- 17465628 TI - Intravenous fluids post marathon : when and why? AB - The medical management of marathon casualties involves several potential treatment pathways. It is helpful to develop defined treatment protocols for commonly experienced conditions addressing intervention selection criteria and monitoring response to therapy. Providing intravenous (IV) fluids for runners post marathon should be scrutinised based upon the effectiveness and safety of the intervention. Commonly agreed upon indications for IV fluids are replacement for clinical dehydration and support for unconscious, hypoglycaemic or persistently hypotensive athletes. Most clinicians recommend serum sodium evaluation prior to IV initiation. IV fluid use in athletes with persistent nausea, generalised muscle cramping and to augment whole-body cooling is commonly deployed despite the paucity of supporting scientific evidence and remains an area for further study. Marathon medical support leaders should develop and disseminate guidelines to direct the administration of IV fluids at their medical aid stations. PMID- 17465629 TI - Epidemiology and aetiology of marathon running injuries. AB - Over the last 10-15 years, there has been a dramatic increase in popularity of running marathons. Numerous articles have reported on injuries to runners of all experience, with yearly incidence rates for injury reported to be as high as 90% in those training for marathons. To date, most of these studies have been cohort studies and retrospective surveys with remarkably few prospective studies. However, from the studies available, it is clear that more experienced runners are less prone to injury, with the number of years running being inversely related to incidence of injuries. For all runners, it is important to be fully recovered from any and all injury or illness prior to running a marathon. For those with less experience, a graduated training programme seems to clearly help prevent injuries with special attention to avoid any sudden increases in running load or intensity, with a particularly high risk for injury once a threshold of 40 miles/week is crossed. In both sexes, the most common injury by far was to the knee, typically on the anterior aspect (e.g. patellofemoral syndrome). Iliotibial band friction syndrome, tibial stress syndrome, plantar fasciitis, Achilles tendonitis and meniscal injuries of the knee were also commonly cited. PMID- 17465630 TI - Exertional heat stroke in the marathon. AB - Exertional heat stroke (EHS) during or following a marathon race can be fatal if not promptly recognised and treated. EHS is a true medical emergency and immediate cooling markedly improves the outcomes. It is critical to recognise EHS and stop the cell damage before the cascade of heat-induced tissue changes becomes irreversible. The goal is to keep the area that is >40.5 degrees C under the body temperature versus time curve at <60 degree-minutes. Measuring the rectal temperature is the only precise estimate of core temperature available for field use. The field treatment of EHS is immediate, total-body cooling with ice water tub immersion or rapidly rotating ice-water towels to the trunk, extremities and head, combined with ice packing of the neck, axillae and groin. Any combination of delayed recognition or cooling increases the potential for morbidity and mortality. For optimal outcomes, it is best to treat immediately with on-site whole-body cooling if cardiorespiratory status is 'stable' and then to transfer the runner for additional evaluation and care. PMID- 17465631 TI - Cardiovascular adaptations to marathon running : the marathoner's heart. AB - Endurance exercise training produces a series of cardiac adaptations including resting bradycardia, first and second degree atrioventricular block, increased intolerance to orthostatic stress, and enlargement of the left ventricular walls and of all cardiac chambers. Cardiac dimensions may be increased beyond the upper limits of normal and some endurance athletes demonstrate mild reductions in estimated left ventricular ejection fraction. Among athletes, such adaptations occur primarily in well trained endurance athletes. Clinicians should be aware of the cardiac changes accompanying endurance training to avoid unnecessary evaluation of physiological changes. On the other hand, the presence of conduction abnormalities or cardiac enlargement in low level or recreational athletes should prompt a search for pathological causes. Many of these findings were presented in the 1977 report on the marathon and have simply been better defined with subsequent studies. PMID- 17465632 TI - Marathon cardiac deaths : the london experience. AB - Data from the London Marathon, with 650,000 completed runs, show that cardiac arrests occur even in the most experienced runners. Although coronary artery disease was the commonest cause of sudden cardiac arrest (SCA) with five deaths and six resuscitations, hypertrophic cardiomyopathy or idiopathic left ventricular hypertrophy (HCM) was diagnosed at autopsy on three occasions. HCM deaths had the same average age as the runners with ischaemic heart disease who had SCA or sudden cardiac death. The cardiac arrests were at the finish in less than one-third of cases and the remainder occurred between 6 and 26 miles on the course. Only one of the eight runners who died had reported symptoms to his family or physician suggestive of cardiac disease. The runner who had reported pre-race angina pain was investigated with a negative exercise stress test prior to the marathon and despite this died with a left anterior descending coronary artery stenosis. The cardiac death rate for the London Marathon is 1 in 80,000 finishers. PMID- 17465633 TI - Hyponatraemia : identification and evaluation in the marathon medical area. AB - Exercise-associated hyponatraemia (EAH) in marathon runners has been reported in the medical literature with incidence varying from 0-130 per 1000 finishers. EAH assessment is enhanced utilising a combination of race weight changes, screening questions, vital signs and clinical examination. Those who fail to lose 0.75kg are seven times more likely to be hyponatraemic than those who lose >0.75kg. Because EAH presentations vary, a three-level surveillance system may help identify mild to moderate cases of EAH that may progress and speed treatment for those in need. After the initial evaluation, the clinical response to treatment is very important to evaluating the severity of EAH. For mild symptomatic hyponatraemia, restrict hypotonic fluids until the runner is urinating and give oral hypertonic solutions if the runner can take oral fluids. For severely symptomatic EAH, intravenous 3% sodium chloride solution will speed recovery and improve outcomes. Pre-race education addressing early symptoms and expected weight changes as well as follow-up instructions detailing the appropriate post race fluids should be available to all registered runners and specifically given to those runners monitored/treated in the medical area. PMID- 17465634 TI - Renal function and vasopressin during marathon running. AB - Over the past 2 decades, exercise-associated hyponatraemia (EAH) has emerged as an important complication of prolonged endurance physical activities. Data collected since the first reports of EAH have strongly implicated a dilutional hyponatraemia from inappropriate retention of body water as the primary cause of EAH. Although high rates of fluid consumption clearly contribute to the pathogenesis of EAH, a review of the available data does not support the view that EAH can be ascribed solely to excess drinking. Because the kidney is exquisitely sensitive to low plasma levels of the antidiuretic hormone arginine vasopressin (AVP) and because many non-osmotic stimuli to AVP secretion normally occur during prolonged endurance exercise activity, it is more likely that a combination of higher than normal fluid intakes in the setting of modest elevations of plasma AVP levels from a variety of potential stimuli during prolonged physical activity accounts for the majority of cases of EAH. In any individual, the degree to which AVP secretion is stimulated and whether it can be suppressed with sufficient fluid ingestion, will determine their susceptibility to EAH as a result of fluid ingestion both before and after physical activity, accounting for the high degree of individual variability in the occurrence of this potentially life-threatening metabolic disorder. PMID- 17465635 TI - Hypertonic (3%) sodium chloride for emergent treatment of exercise-associated hypotonic encephalopathy. AB - Exercise-associated hyponatraemia (EAH) is an acute-onset imbalance in the tonicity of extracellular fluids during or after endurance exercise which results in a blood sodium concentration <135 mmol/L. Both excessive fluid intake and a concurrent decrease in urine formation contribute to this rapid-onset, predominantly dilutional, decrease in serum sodium, which can result in life threatening pulmonary and cerebral oedema. Marathon runners with hypotonic encephalopathy related to EAH, including two cases with fatal cerebral oedema, demonstrated non-osmotic secretion of arginine vasopressin and fulfilled the essential diagnostic criteria for the syndrome of inappropriate antidiuretic hormone secretion (SIADH). The pathophysiology of SIADH as the proximate cause of EAH accounts for otherwise puzzling clinical observations such as cases occurring after only moderate fluid intake or presenting hours after races. This formulation provides a framework for enhancing prevention by monitoring weight changes during races to detect positive fluid balance before the onset of mental status changes. Most importantly, SIADH supports a strategy for use of oral and intravenous hypertonic solutions, including 3% sodium chloride, for the emergent treatment of moderate and life-threatening symptoms of hypotonic encephalopathy, respectively. PMID- 17465636 TI - Hydration in the marathon : using thirst to gauge safe fluid replacement. AB - Early human ancestors evolved hunting in the midday heat on the dry African savannah and developed favourable biological adaptations that permit prolonged running in the heat. These physiological adaptations must have included the capacity to sweat profusely to maintain a low body temperature when running for 4 6 hours in dry heat, an absence of adverse consequences from developing mild to moderate fluid deficits caused by sweat losses during the hunt, a serum osmolality based thirst mechanism and the ability to 'outrun their thirst' (to resist the deleterious psychological and other effects of severe thirst). Until the early 1970s, the guidelines for fluid ingestion during exercise were not to drink and are consistent with this interpretation. By 1996, guidelines stated, "individuals should be encouraged to consume the maximal amount of fluids during exercise that can be tolerated without gastrointestinal discomfort up to a rate equal to that lost from sweating," and this was interpreted by some as "to drink as much as tolerable." This article argues that humans are designed to drink just enough to maintain plasma osmolality, not necessarily bodyweight, both at rest and during exercise. Drinking to maintain bodyweight may impair exercise performance by inducing a weight penalty and may increase the probability of exercise-associated hyponatraemia in slow marathon runners. PMID- 17465637 TI - Clinical pharmacokinetics of atypical antipsychotics: a critical review of the relationship between plasma concentrations and clinical response. AB - In the past, the information about the dose-clinical effectiveness of typical antipsychotics was not complete and this led to the risk of extrapyramidal adverse effects. This, together with the intention of improving patients' quality of life and therapeutic compliance, resulted in the development of atypical or second-generation antipsychotics (SGAs). This review will concentrate on the pharmacokinetics and metabolism of clozapine, risperidone, olanzapine, quetiapine, amisulpride, ziprasidone, aripiprazole and sertindole, and will discuss the main aspects of their pharmacodynamics. In psychopharmacology, therapeutic drug monitoring studies have generally concentrated on controlling compliance and avoiding adverse effects by keeping long-term exposure to the minimal effective blood concentration. The rationale for using therapeutic drug monitoring in relation to SGAs is still a matter of debate, but there is growing evidence that it can improve efficacy, especially when patients do not respond to therapeutic doses or when they develop adverse effects. Here, we review the literature concerning the relationships between plasma concentrations of SGAs and clinical responses by dividing the studies on the basis of the length of their observation periods. Studies with clozapine evidenced a positive relationship between plasma concentrations and clinical response, with a threshold of 350-420 ng/mL associated with good clinical response. The usefulness of therapeutic drug monitoring is well established because high plasma concentrations of clozapine can increase the risk of epileptic seizures. Plasma clozapine concentrations seem to be influenced by many factors such as altered cytochrome P450 1A4 activity, age, sex and smoking. The pharmacological effects of risperidone depend on the sum of the plasma concentrations of risperidone and its 9-hydroxyrisperidone metabolite, so monitoring the plasma concentrations of the parent compound alone can lead to erroneous interpretations. Despite a large variability in plasma drug concentrations, the lack of studies using fixed dosages, and discrepancies in the results, it seems that monitoring the plasma concentrations of the active moiety may be useful. However, no therapeutic plasma concentration range for risperidone has yet been clearly established. A plasma threshold concentration for parkinsonian side effects has been found to be 74 ng/mL. Moreover, therapeutic drug monitoring may be particularly useful in the switch between the oral and the long-acting injectable form. The reviewed studies on olanzapine strongly indicate a relationship between clinical outcomes and plasma concentrations. Olanzapine therapeutic drug monitoring can be considered very useful in assessing therapeutic efficacy and controlling adverse events. A therapeutic range of 20-50 ng/mL has been found. There is little evidence in favour of the existence of a relationship between plasma quetiapine concentrations and clinical responses, and an optimal therapeutic range has not been identified. Positron emission tomography studies of receptor blockade indicated a discrepancy between the time course of receptor occupancy and plasma quetiapine concentrations. The value of quetiapine plasma concentration monitoring in clinical practice is still controversial. Preliminary data suggested that a therapeutic plasma amisulpride concentration of 367 ng/mL was associated with clinical improvement. A therapeutic range of 100-400 ng/mL is proposed from non-systematic clinical experience. There is no direct evidence concerning optimal plasma concentration ranges of ziprasidone, aripiprazole or sertindole. PMID- 17465638 TI - Pharmacokinetics of mycophenolic acid and estimation of exposure using multiple linear regression equations in Chinese renal allograft recipients. AB - OBJECTIVES: To investigate the pharmacokinetics of mycophenolic acid (MPA) in Chinese adult renal allograft recipients, and to generate the validated model equations for estimation of the MPA area under the plasma concentration-time curve from 0 to 12 hours (AUC(12)) with a limited sampling strategy. PATIENTS AND METHODS: The pharmacokinetics in 75 Chinese renal allograft recipients treated with mycophenolate mofetil 2 g/day in combination with cyclosporin and corticosteroids were determined. The MPA concentration was assayed by high performance liquid chromatography at pre-dose (C(0)) and at 0.5 (C(0.5)), 1 (C(1)), 1.5 (C(1.5)), 2 (C(2)), 4 (C(4)), 6 (C(6)), 8 (C(8)), 10 (C(10)) and 12 (C(12)) hours after dosing on day 14 post-transplant. Patients were randomly divided into: (i) a model group (n = 50) to generate the model equations by multiple stepwise regression analysis for estimation of the MPA AUC by a limited sampling strategy; and (ii) a validation group (n = 25) to evaluate the predictive performance of the model equations. RESULTS: The mean MPA AUC(12) was 52.97 +/- 15.09 mg . h/L, ranging from 24.0 to 102.3 mg . h/L. The patient's age and serum albumin level had a significant impact on the MPA AUC(12). The correlation between the pre-dose MPA trough level (C(0)) and the MPA AUC(12) was poor (r(2) = 0.02, p = 0.33). Model equations 7 (MPA AUC(12) = 14.81 + 0.80 . C(0.5) + 1.56 . C(2) + 4.80 . C(4), r(2) = 0.70) and 11 (MPA AUC(12) = 11.29 + 0.51 . C(0.5) + 2.13 . C(2) + 8.15 . C(8), r(2) = 0.88) were selected for MPA AUC calculation in Chinese patients, resulting in good agreements between the estimated MPA AUC and the full MPA AUC(12), with a mean prediction error of +/ 10.1 and +/-6.9 mg . h/L, respectively. CONCLUSION: In Chinese renal allograft recipients, MPA pharmacokinetics manifest substantial interindividual variability, and the MPA AUC(12) tends to be higher than that in Caucasian patients receiving the same dose of mycophenolate mofetil. Two validated model equations with three sampling timepoints are recommended for MPA AUC estimation in Chinese patients. PMID- 17465639 TI - Steady-state pharmacokinetics of roflumilast and roflumilast N-oxide in patients with mild and moderate liver cirrhosis. AB - BACKGROUND: Roflumilast and its primary N-oxide metabolite are targeted phosphodiesterase 4 (PDE4) inhibitors with similar in vivo potency. Roflumilast is being developed for the treatment of inflammatory airway diseases such as chronic obstructive pulmonary disease and asthma. OBJECTIVE: To investigate the effects of mild and moderate liver cirrhosis on the steady-state pharmacokinetics of roflumilast and roflumilast N-oxide. METHODS: Patients with mild (n = 8, Child Pugh A) and moderate (n = 8, Child-Pugh B) liver cirrhosis and healthy subjects (n = 8) matched with patients with cirrhosis with regard to sex, age and bodyweight received oral roflumilast 250 microg once daily for 14 days. Blood samples were collected for 24 hours after the last dose on day 14. Steady-state plasma concentrations of roflumilast and roflumilast N-oxide were determined using a validated high-performance liquid chromatography with tandem mass spectrometry assay. The pharmacokinetics were compared between groups using ANOVA. RESULTS: In patients with liver cirrhosis, the average total exposure (area under the plasma concentration-time curve from 0 to 24 hours [AUC(24)]) of roflumilast was approximately 51% (Child-Pugh A) and 92% (Child-Pugh B) higher than in healthy subjects. In contrast, roflumilast maximum plasma concentration (C(max)) was unaltered in Child-Pugh A patients and was increased by 27% in Child Pugh B patients. Changes in the AUC(24) of roflumilast N-oxide were less distinct, with 24% and 41% increases and corresponding C(max) increases of 26% and 40% in Child-Pugh A and B patients, respectively, compared with healthy subjects. Overall, changes in average potency-corrected exposure to the sum of the free fractions of both compounds were estimated to result in approximately 26% and 46% increases in total PDE4 inhibitory capacity (tPDE4i) in Child-Pugh A and B patients, respectively, relative to healthy subjects. Roflumilast was well tolerated. CONCLUSIONS: Mild and moderate liver cirrhosis resulted in distinct alterations of exposure to roflumilast but only in modest alterations of exposure to roflumilast N-oxide. The integrated exposure-weighted assessment of the observed pharmacokinetic changes of roflumilast and roflumilast N-oxide (tPDE4i) indicates modest average exposure increases to the sum of both compounds. These findings and the favourable tolerability profile suggest that roflumilast can be safely used in patients with mild and moderate liver cirrhosis without special precautions or dose adjustment. PMID- 17465640 TI - Exposure-effect population model of inolimomab, a monoclonal antibody administered in first-line treatment for acute graft-versus-host disease. AB - BACKGROUND AND OBJECTIVE: Inolimomab, a monoclonal antibody against interleukin (IL)-2Ralpha (CD25) has shown promising results in the treatment of corticosteroid-resistant acute graft-versus-host disease (GvHD). The objective of the present study was to characterise the pharmacokinetic and pharmacodynamic properties of inolimomab as first-line treatment in this condition. METHODS: The data came from 21 patients with acute GvHD (8 with an International Bone Marrow Transplant Registry [IBMTR] score of B, 11 with a score of C and 2 with a score of D) following haematopoietic stem cell transplantation after a median delay of 26 days (range 10-127 days). Inolimomab was administered at 0.1, 0.2, 0.3 or 0.4 mg/kg daily in association with methylprednisolone (2 mg/kg) for 8 or 16 days depending on the status at day 9. Then, for responder patients, administrations were continued three times weekly until day 28. Inolimomab concentrations and pharmacodynamic data (acute GvHD scores) were recorded during the study. The pharmacodynamic data were assessed in four grades according to the IBMTR and Glucksberg classification in parallel with Karnofsky scores. A population analysis was developed using a nonlinear mixed-effects model to define the pharmacokinetic model, to test covariates and, when apparent, to model the exposure-effect relationship by a proportional odds model. The modelling was finally qualified by a predictive check. RESULTS: The best pharmacokinetic model was two-compartmental. For each score, the most demonstrative exposure-effect graphics linked the cumulative area under the concentration-time curve to cumulated probabilities of observing a specific score. This relationship was identified as a maximum effect model for the skin (with two patient subpopulations: sensitive/less sensitive) and a linear model for the intestinal tract and liver. No covariate was identified as influencing any of these parameters. CONCLUSION: Inolimomab exposure-effect relationships as first-line treatment for acute GvHD have been identified and modelled. The discovered dose effect relationship allows confirmation of the treatment response, thereby establishing the first step towards optimising the inolimomab dosage in future trials. PMID- 17465641 TI - Animal-to-human extrapolation of the pharmacokinetic and pharmacodynamic properties of buprenorphine. AB - OBJECTIVES: This investigation describes the interspecies scaling of the pharmacokinetics and pharmacodynamics of buprenorphine. METHODS: Data on the time course of the antinociceptive and respiratory depressant effects of buprenorphine in rats and in humans were simultaneously analysed on the basis of a mechanism based pharmacokinetic-pharmacodynamic model. RESULTS: An allometric three compartment pharmacokinetic model described the time course of the concentration in plasma. The value of the allometric coefficient for clearance was 35.2 mL/min (relative standard error [RSE] = 5.6%) and the value of the allometric exponent was 0.76 (RSE 5.61%). A combined biophase distribution-receptor association/dissociation model with a linear transduction function described hysteresis between plasma concentration and effect. The values of the drug specific pharmacodynamic parameters were identical in rats and in humans. For the respiratory depressant effect, the values of the second-order rate constant of receptor association (k(on)) and the first-order rate constant of receptor dissociation (k(off)) were 0.23 mL/ng/min (RSE = 15.8%) and 0.014 min(-1) (RSE = 27.7%), respectively, and the value of the equilibrium dissociation constant (K(diss)) was 0.13 nmol/L. The value of the intrinsic activity alpha was 0.52 (RSE = 3.4%). For the antinociceptive effect, the values of the k(on) and k(off) were 0.015 mL/ng/min (RSE = 18.3%) and 0.053 min(-1) (RSE = 23.1%), respectively. The value of the K(diss) was 7.5 nmol/L. An allometric equation described the scaling of the system-specific parameter, the first-order distribution rate constant (k(e0)). The value of the allometric coefficient for the k(e0) was 0.0303 min(-1) (RSE = 11.3%) and the value of the exponent was -0.28 (RSE = 9.6%). CONCLUSIONS: The different values of the drug-specific pharmacodynamic parameters are consistent with the different opioid mu receptor subtypes involved in the antinociceptive and respiratory depressant effects. PMID- 17465643 TI - Long-term soft tissue changes after orthodontic and surgical corrections of skeletal class III malocclusions. AB - OBJECTIVE: To evaluate long-term soft tissue changes after orthodontic and surgical corrections of skeletal Class III malocclusions. MATERIALS AND METHODS: Postoperative cephalometric radiographs at 1 year and at 5 years or more after treatment were digitized for 92 patients who had surgical correction of their Class III problem by LeFort I maxillary advancement (n = 48), mandibular setback (n = 12), or a combination of the two procedures (n = 32) and for 25 patients who received orthodontic treatment only. RESULTS: For all groups, the mean changes were quite small. For most measurements, fewer than 20% of patients experienced long-term changes from 2 mm to 4 mm, and fewer than 10% experienced long-term changes greater than 4 mm. CONCLUSIONS: A smaller percentage of surgically treated Class III patients showed long-term soft tissue changes than did surgically treated Class II patients, but compared with both Class II patients and untreated adults they experienced greater long-term forward projection of the soft tissue chin. PMID- 17465644 TI - Longitudinal intermaxillary relationships in class III malocclusions with low and high mandibular plane angles. AB - OBJECTIVE: To analyze the sagittal, vertical, and transverse relationships of the maxilla and mandible in Japanese girls with Class III malocclusions with different inclination of mandibular plane. MATERIALS AND METHODS: This longitudinal study utilized serial posteroanterior and lateral cephalograms of 56 untreated subjects from the age of 8 years until the age of 14 years (low mandibular plane angle group: n = 20; average mandibular plane angle group: n = 15; high mandibular plane angle group: n = 21). Sagittal and vertical growth was analyzed on the basis of lateral cephalograms, and transverse growth was analyzed on the basis of posteroanterior cephalograms. RESULTS: There was no significant difference in intermaxillary sagittal relationships among the three groups from age 8 until 14. On the other hand, there were significant changes in the vertical and transverse intermaxillary relationships during this period. When comparing the three groups at the same age, there were significant differences in vertical and transverse intermaxillary relationships in some ages, whereas no significant difference was found in sagittal relationships in any ages. CONCLUSIONS: The inclination of mandibular plane might play a role in anticipating changes in vertical and transverse intermaxillary relationships from 8 until 14 years of age. PMID- 17465645 TI - Orthodontic space opening in patients with congenitally missing lateral incisors. AB - OBJECTIVE: To determine the best time to begin orthodontic treatment for patients scheduled for implants to replace congenitally missing upper lateral incisors. The aim of timing is to maximize the amount of bone available for implant insertion and to improve incisors inclination. MATERIALS AND METHODS: Seventy three plaster casts of 14 patients with 26 missing lateral incisors were cross sectioned in the center of the planned insertion of the implant, and the implant profile was projected into the area at three different times: T1--beginning of orthodontic treatment, T2-- end of orthodontic treatment, and T3--implant insertion. Deficiency of alveolar ridge volume needed for implantation was determined by Leica Quin analySIS software. RESULTS: An increase of ridge-volume deficiency from 0.26 mm(2) at T1 to 3.77 mm(2) at T3 was found. During orthodontic treatment the incisors protruded about 9.4 degrees (differing from the O1-NA standard of 7.5 degrees ). To ensure optimal esthetic and functional implantation results, time management concerning orthodontic treatment has to be done carefully. CONCLUSIONS: To avoid a high degree of alveolar bone atrophy and the risk of relapse and retreatment, orthodontic treatment involving tooth movement should not be initiated before the age of 13 years. Furthermore, it is important to maintain anchorage of the upper incisors because high incisor proclination causes extra-axial stress on the implant. An interdisciplinary approach is essential to provide the best treatment outcome. PMID- 17465646 TI - Histological evaluation of the effects of initially light and gradually increasing force on orthodontic tooth movement. AB - OBJECTIVE: To investigate histologically the effect of initially light and gradually increasing force on tooth movement in the rat. MATERIALS AND METHODS: Cuboids made of neodymium-iron-boron magnets (experimental groups) or titanium (control group) were bonded to the lingual surface of the right and left maxillary first molars of 18-week-old male Wistar rats. The initial distances between materials were 1.0 mm generating 4.96 gf (experimental group 1) and 1.5 mm generating 2.26 gf (experimental group 2). In three groups, rats were killed 1, 3, 7, 10, or 14 days after treatment. Histological sections were prepared and stained with hematoxylin and eosin or for tartrate-resistant acid phosphatase (TRAP) activity. The number of TRAP-positive osteoclasts was counted, and the relative hyalinized area was measured on the pressure side of periodontal ligament. RESULTS: There were significant differences in the number of osteoclasts among the three groups (P < .05). On days 1 and 3, the numbers of osteoclasts in experimental group 2 were greater than in experimental group 1. There were significant differences in the relative hyalinized area between the control group and experimental group 1 (P < .01) and between experimental groups 1 and 2 (P < .01). On days 1 and 3, the hyalinized area in experimental group 1 was larger than in experimental group 2. CONCLUSION: Initially light and gradually increasing force induced tooth movement without the lag phase and showed smooth recruitment of osteoclasts and inhibition of hyalinization. PMID- 17465647 TI - Orthodontic treatment acceleration with corticotomy-assisted exposure of palatally impacted canines. AB - OBJECTIVE: To evaluate the effectiveness of a new surgical technique in the treatment of palatally impacted canines. MATERIALS AND METHODS: Six consecutive patients presenting with bilaterally impacted canines were compared. One canine was surgically exposed using a conventional surgical technique while the contralateral canine was exposed using a corticotomy-assisted technique. RESULTS: After tooth movement was completed, statistical comparisons of the two methods revealed a reduction of treatment time of 28-33% for the corticotomy-assisted canines. No significant differences were observed in final periodontal condition between the canines exposed by these two methods. CONCLUSION: This preliminary study supports the concept that a corticotomy-assisted surgical technique helps reduce orthodontic treatment time for palatally impacted canines. PMID- 17465648 TI - The dentition and occlusal development in children of african american descent. AB - OBJECTIVE: The objectives of this study were to (1) determine normative dental arch dimensions (arch length [AL], arch width [AW], arch perimeter [AP], and amount of interdental space [IDS]) in the primary dentition of African American (AA) children; (2) compare the established normative arch dimensions in AA children with their European American (EA) cohorts; and (3) contrast a theoretical construct of mandibular transitional occlusal development between children of AA and EA descent on the basis of normative values. MATERIALS AND METHODS: Two hundred seventeen sets of AA primary dental casts (110 boys, 107 girls) made from alginate impressions were measured and compared with historical EA standard values. Independent group t-tests were used to analyze sample differences. RESULTS: AA children showed significant (P = .001) sex differences in each respective dental arch dimension with the exception of the mandibular canine AW. AA boys and girls revealed statistically larger (P = .001) arch dimensions (AP, AL, AW) and amounts of IDS compared with their EA cohorts. CONCLUSIONS: The total amount of IDS within the primary dental arches is approximately equal in AA boys and girls, but significant site-specific gender dimorphism existed. The primary dental arch dimensions of AA children were significantly larger than those of EA children in AW, AL, AP, and IDS. PMID- 17465649 TI - Maxillary transverse discrepancies and potentially impacted maxillary canines in mixed-dentition patients. AB - OBJECTIVE: To investigate the correlation between maxillary transverse discrepancy and the occurrence of impacted canines in patients during the mixed dentition stage. MATERIALS AND METHODS: Panoramic radiographs and dental casts were evaluated of randomly selected patients in the mixed dentition. The experimental group consisted of 84 orthodontic patients with a maxillary transverse discrepancy. The control group included 100 orthodontic patients without a maxillary transverse discrepancy. Intermolar widths of the experimental group were measured and recorded. The permanent canines of both groups were placed into a sector classification by using a panoramic radiograph. The experimental group was then analyzed to identify whether these patients had an impacted maxillary canine associated with the transverse discrepancy. The results were further evaluated based on type of impaction (unilateral or bilateral). RESULTS: Results of this study showed that patients with a transverse discrepancy are more likely to have an impacted canine than those patients without a transverse discrepancy, with the impaction more likely being unilateral. However, patients with a transverse discrepancy do not have a greater likelihood of having a bilateral impaction compared with patients without a transverse discrepancy. CONCLUSIONS: There appears to be an association between potentially impacted canines and transverse discrepancies. Identification can be made early based on proper panoramic evaluation and clinical detection. If a possibly impacted canine is detected early, appropriate treatment should be taken to minimize complications and avoid definitive impaction. PMID- 17465650 TI - Functional changes of the temporomandibular joint mechanoreceptors induced by a lateral mandibular shift in rats. AB - OBJECTIVE: To investigate changes in functional characteristics of the temporomandibular joint (TMJ) mechanoreceptors under mandibular lateral shift. MATERIALS AND METHODS: Forty 7-week-old male Wistar rats were divided into control and experimental groups. Rats in the experimental group received a modified guiding appliance (MGA) that was designed to shift the mandible to the left side in the occlusal position. Single-unit activities of the TMJ mechanoreceptors were evoked by indirect stimulation of passive jaw movement. Electrophysiologic recordings of TMJ units were obtained 1, 3, 5, 7, and 9 weeks after MGA setting from the gasserian ganglion. RESULTS: At 1 week after mandibular shift, the firing thresholds were the lowest and the maximum instantaneous frequencies were the highest in TMJ units. At 5 weeks, the firing thresholds in the nonshifted side were significantly lower than those in the shifted side. The maximum instantaneous frequencies in the nonshifted side were significantly higher than those in the shifted side at 1, 5, and 7 weeks. CONCLUSION: These results suggest that the functional lateral shift of the mandible could alter the response properties of TMJ mechanoreceptors, particularly on the nonshifted side. PMID- 17465651 TI - Longitudinal effects of rapid maxillary expansion. AB - OBJECTIVE: To evaluate the long-term effects of rapid maxillary expansion (RME) via banded expanders in the sagittal and vertical facial planes. MATERIALS AND METHODS: The sample consisted of 25 patients who had undergone RME (with either Haas-type or Hyrax hygienic expanders) followed by standard edgewise orthodontic therapy. This sample was compared with a group of 25 patients who had edgewise treatment only and with a control nontreatment group of 26 subjects, matched by age and gender with the patients of the other two groups. Lateral cephalograms were taken before treatment (T1), at the end of treatment (T2), and at 3 years posttreatment (T3), comprising a 5-year average time of observation. RESULTS: RME treatment, in the long-term, did not influence the sagittal position of the apical jaw bases or the facial vertical dimension. CONCLUSION: Unfavorable cephalometric changes resulting immediately after RME are temporary, and therefore concerns about using RME in patients with vertical growth patterns or an extremely convex facial profile are not substantiated. PMID- 17465652 TI - Treatment efficiency in skeletal Class II patients treated with the jasper jumper. AB - OBJECTIVE: To analyze the effects of the Jasper Jumper appliance during the treatment of skeletal Class II malocclusion. MATERIALS AND METHODS: Lateral cephalograms and hand-wrist radiographs were collected from 45 Class II growing patients (22 boys, 23 girls). Three sets of records (initial, before Jasper Jumper, after Jasper Jumper) from 25 patients were compared with 20 control subjects of similar skeletal developmental stage. Mean age of the treatment and control groups were 11.83 years and 11.3 years, respectively. The data were analyzed by using paired t-tests. RESULTS: The results demonstrated that the Jasper Jumper effectively corrected Class II malocclusion, but the changes were 80% dentoalveolar. The Jasper Jumper induced a clockwise rotation of the occlusal plane without much alteration in vertical dimension. Skeletally, the maxillary growth was restricted and pogonion moved forward, improving the profile. CONCLUSION: The Jasper Jumper appliance may be an effective method to improve both the skeletal imbalance and the profile in growing patients. PMID- 17465653 TI - Masticatory exercise as an adjunctive treatment for hyperdivergent patients. AB - OBJECTIVE: This retrospective study was designed to evaluate the morphologic effects of masticatory muscle exercise as an adjunctive therapy for hyperdivergent patients treated with fixed orthodontic appliances. MATERIALS AND METHODS: Three samples of 50 subjects were selected, including one sample treated with orthodontics combined with exercise, one sample treated with orthodontics only, and an untreated control sample. Subjects were matched on the basis of age, sex, mandibular plane angle (MPA), treatment duration, and treatment rendered. Patients in the treated exercise sample were instructed to clench their teeth together as hard as possible for 15 seconds and to repeat this process at least four times for a total of one minute; this one-minute exercise was to be performed as often as possible throughout the day. Morphologic data was derived from pre- and posttreatment lateral cephalograms. RESULTS: Exercise with orthodontics produced significant (P < .05) increases in overbite compared to orthodontics alone. However, changes in vertical facial morphology were not significantly different between the two treated samples. Relative to untreated controls, both treated samples showed significantly greater increases in the MPA (S-N to Go-Me), Y-axis, and the lower to total facial height ratio; the treated samples also showed significantly less true forward mandibular rotation than the untreated controls. CONCLUSION: Short-term clenching exercises performed daily are insufficient as an adjunct to traditional orthodontic treatment for correcting or controlling the vertical dimension. PMID- 17465654 TI - Skeletal sagittal and vertical facial types and electromyographic activity of the masticatory muscle. AB - OBJECTIVE: To investigate the electromyographic activities of the anterior temporal (T) and masseter (M) muscles in different facial skeletal types. MATERIALS AND METHODS: The samples consisted of 105 subjects (38 males and 67 females; mean age 22.0 +/- 6.7 years) and were classified into six groups according to the values of ANB and SN-GoMe: group 1 for Class I malocclusion and normodivergent type (n = 27), group 2 for Class I and hyperdivergent type (n = 20), group 3 for Class II and normodivergent type (n = 10), group 4 for Class II and hyperdivergent type (n = 23), group 5 for Class III and normodivergent type (n = 12), and group 6 for Class III and hyperdivergent type (n = 13). Temporal muscle activity (TMA), masseter muscle activity (MMA), and T/M ratio were evaluated at resting and clenching status. RESULTS: Although there was no significant difference in resting MMA among all groups, group 6 showed a higher resting TMA than did other groups and a significant difference in resting T/M ratio compared with groups 1 and 3. There were no significant differences in clenching TMA and MMA among all groups. Although all groups showed a significant increase of TMA and MMA from resting to clenching status, group 6 showed a significant decrease of clenching T/M ratio compared with resting T/M ratio. CONCLUSIONS: The results suggest that the more Class III and the more hyperdivergent type, the higher resting TMA and the lesser increase of clenching MMA than expressed by other groups. Significant differences existed in TMA and MMA according to sagittal and vertical facial skeletal types. PMID- 17465655 TI - Occlusal risk factors for temporomandibular disorders. AB - OBJECTIVE: To determine the role of occlusal variables (overbite; overjet; number of anterior and posterior teeth; bilateral canine guidance on lateral and protrusive movements; anterior centric slide; Angle Classes I, II, and III malocclusion) as risk indicators for the development of temporomandibular disorders (TMDs). MATERIALS AND METHODS: Seventy-two TMD patients with myofascial pain, with or without limited opening and arthralgia, as well as 30 age- and gender-matched pain-free concurrent controls were included. The association (critical odds ratio [OR] = 2.0) between the significant occlusal variables and TMD was calculated. Confounders were controlled in the inclusion-exclusion criteria as well as in the analysis stage (unconditional logistic regression) by variation in the OR (15%). RESULTS: Angle Class II malocclusion (crude OR = 8.0, confidence interval [CI] = 2.2 to 29.3) and the absence of bilateral canine guidance on lateral excursion (crude OR = 3.9, CI = 1.6 to 9.7) were statistically more common in patients than in controls. Spontaneous pain as well as pain on palpation (Class II or higher) were also statistically worse in TMD patients. Significant confounders (ie, employment, age, cigarette and alcohol consumption) acted as effect modifiers not changing the critical OR (adjusted OR Angle Class II and bilateral canine guidance = 8.3 to 12.4 and 2.2 to 4.1, respectively). CONCLUSIONS: Absence of bilateral canine guidance on lateral excursion and particularly Angle Class II malocclusion were considered important risk indicators for the development of TMD in this investigation, even when some sociodemographic factors were considered as effect modifiers. PMID- 17465656 TI - Temperature rise in the pulp chamber during different stripping procedures. AB - OBJECTIVE: To measure the temperature changes in the pulp chamber when different stripping procedures were used without any type of coolant. MATERIALS AND METHODS: Ninety intact, freshly extracted human teeth were used in this study. The teeth were separated into nine groups of 10 teeth each. Mesial and distal sides of the teeth were used separately. The stripping procedures were performed on three different tooth groups (incisor, canine, premolar) with a metal handheld stripper, perforated stripping disk, or tungsten carbide bur. A J-type thermocouple wire was positioned in the center of the pulp chamber and was connected to a data logger during application of stripping procedures. The results were analyzed by analysis of variance (ANOVA) and the Duncan test. RESULTS: Two-factor ANOVA revealed significant interaction between the stripping procedure and the tooth type (P = .000). The results of this study demonstrate that tungsten carbide burs used on mandibular incisors had the highest temperature variation (DeltaT) values, which exceeded the critical level (5.5 degrees C), and this was significantly higher than those of the other stripping procedures (DeltaT: 5.63 +/- 1.73 degrees C). On the other hand, six of the nine groups also produced temperature increases above the critical level (5.5 degrees C) for some of the specimens. CONCLUSIONS: Frictional heat is a common side effect of stripping procedures, and appropriate measures (ie, cooling application) should be taken particularly for high-speed hand-piece stripping of mandibular incisors. PMID- 17465657 TI - Surface modification of orthodontic wires with photocatalytic titanium oxide for its antiadherent and antibacterial properties. AB - OBJECTIVE: To test the antiadherent and antibacterial properties of surface modification of orthodontic wires with photocatalytic titanium oxide (TiO(2)). MATERIALS AND METHODS: TiO(2) was coated on the surface of the orthodontic wires by a sol-gel thin film dip-coating method. Bacterial adhesion to the wires was evaluated by the weight change of the wires. The antibacterial activity of the surface-modified orthodontic wires was demonstrated by the dilution agar plate method for Streptococcus mutans and spectrophotometry for Porphyromonas gingivalis. RESULTS: The orthodontic wires coated with the photocatalytic TiO(2) showed an antiadherent effect against S. mutans compared with the uncoated wires. The bacterial mass that bound to the TiO(2)-coated orthodontic wires remained unchanged, whereas that of the uncoated wires increased by 4.97%. Furthermore, the TiO(2)-coated orthodontic wires had a bactericidal effect on S. mutans and P. gingivalis, which cause dental caries and periodontitis, respectively. The antiadherent and antibacterial mechanisms of TiO(2) to break down the cell wall of those bacteria were revealed by scanning electron microscopy. CONCLUSION: The surface modification of orthodontic wires with photocatalytic TiO(2) can be used to prevent the development of dental plaque during orthodontic treatment. PMID- 17465658 TI - Bleaching and desensitizer application effects on shear bond strengths of orthodontic brackets. AB - OBJECTIVE: To evaluate the effects of bleaching and desensitizer application on shear bond strengths of orthodontic brackets. MATERIALS AND METHODS: Forty-eight extracted human premolar teeth were randomly assigned to 4 groups of 12 each. The first group of teeth was bleached with a 35% hydrogen peroxide office bleaching agent. The second group was bleached the same as the first group and UltraEZ desensitizer was applied. No bleaching procedures were applied on the third and fourth groups. UltraEZ desensitizer alone was applied to teeth in the third group. The fourth group served as control. Orthodontic brackets were bonded with a LC (light cure) composite resin and cured with a halogen light. After bonding, the shear bond strengths of the brackets were tested with a Universal testing machine. RESULTS: The results showed that bleaching, bleaching plus desensitizer, and desensitizer procedures significantly reduced the bonding strengths of the orthodontic brackets (P < .05, P < .001, and P < .01, respectively). No statistically significant difference was found between bleaching, bleaching plus desensitizer, and desensitizer groups (P > .05). CONCLUSIONS: Because bleaching and desensitizer application significantly affected shear bond strengths of orthodontic brackets on human enamel, they should be delayed until the completion of orthodontic treatment. PMID- 17465659 TI - Influence of filler level on the bond strength of orthodontic adhesives. AB - OBJECTIVE: To evaluate the effect of different filler contents of orthodontic adhesives on shear bond strength and Adhesive Remnant Index (ARI). MATERIALS AND METHODS: Four experimental adhesive groups were created: group 1 was an unfilled urethane-dimethacrylate (UDMA) adhesive, group 2 consisted of UDMA and a filler content of 30 vol%, group 3 consisted of UDMA and a filler content of 50 vol%, and group 4 was manufactured with a filler level of 70 vol% in a UDMA matrix. The embedded filler was silicon dioxide. After etching and priming, stainless steel brackets were bonded to extracted human third molars (n = 60) with the experimental adhesives (15 teeth per group). After storage in distilled water for 72 hours, shear bond strength and ARI scores were evaluated. Kruskal-Wallis and post hoc test were performed for statistical analysis. RESULTS: Shear bond strength of UDMA-based adhesives depends on filler content. Higher adhesive filler levels reveal greater bond strength between enamel and stainless steel brackets. ARI scores showed no significant difference among the groups. However, higher filled adhesives (filler content 70 vol%) seem to present higher ARI scores. CONCLUSION: Highly filled UDMA adhesives offer greater bond strength than do lower filled or unfilled resins. PMID- 17465660 TI - Thermal analysis of as-received and clinically retrieved copper-nickel-titanium orthodontic archwires. AB - OBJECTIVE: To compare as-received copper-nickel-titanium (CuNiTi) archwires to those used in patients by means of differential scanning calorimetry (DSC). Also, the thermal or phase properties of 27 degrees C, 35 degrees C, and 40 degrees C CuNiTi archwires were studied to ascertain if their properties match those indicated by the manufacturer. MATERIALS AND METHODS: Six wires of 27 degrees C, 35 degrees C, and 40 degrees C CuNiTi were tested as-received, and six each of the 27 degrees C and 35 degrees C wires were examined after use in patients for an average of approximately 9 and 7 weeks, respectively. Segments of archwire were investigated by DSC over the temperature range from -100 degrees C to 150 degrees C at 10 degrees C per minute. RESULTS: There were no significant differences between as-received and clinically used 27 degrees C and 35 degrees C wires for all parameters (heating onset, endset, and enthalpy and cooling onset, endset, and enthalpy), except the 27 degrees C wires exhibited a significant decrease in the heating enthalpy associated with the martensite-to-austenite transition after clinical use. The heating endsets (austenite finish temperatures) of the 27 degrees C and 35 degrees C wires were within 2 degrees C of those claimed by the manufacturer, but the 40 degrees C wires were found to be nearer to 36 degrees C than 40 degrees C. CONCLUSIONS: Clinical use of CuNiTi wires resulted in few differences when compared with as-received wires analyzed by DSC. Two temperature varieties of CuNiTi are reasonably within the parameters of those identified by the manufacturer. PMID- 17465661 TI - The effect of modifying the self-etchant bonding protocol on the shear bond strength of orthodontic brackets. AB - OBJECTIVE: To compare the shear bond strength (SBS) of orthodontic brackets when the self-etching primer (SEP) and the bracket adhesive are light cured either separately or simultaneously. MATERIALS AND METHODS: Seventy-five human molars were randomly divided into five equal groups. Brackets precoated with Transbond XT composite adhesive were used. The five protocols were: Group 1 (control), the SEP Transbond Plus was applied, brackets placed, and adhesive light cured for 20 seconds; Group 2, SEP Adper Prompt L-Pop was applied, light cured, brackets placed, and light cured; Group 3, the same SEP as in Group 2 was used, however, the SEP and bracket adhesive were light cured together; Group 4, SEP Clearfil S3 Bond was applied, light cured, brackets placed, and light cured; and Group 5, the same SEP as in group 4 was used, however, the SEP and the adhesive were light cured together. The teeth were debonded using a universal testing machine, and the enamel was examined for residual adhesive. Analysis of variance was used to compare the SBS. RESULTS: The SBS of Clearfil S3 Bond after one light cure and two light cures were significantly greater than the bonds of brackets using Transbond Plus. Brackets bonded using Adper Prompt L-Pop after one light cure and two light cures were not significantly different from the other groups. The groups did not differ significantly in their bracket failure modes. CONCLUSION: Only one light curing application is needed to successfully bond brackets when using SEPs and adhesives. This approach can potentially reduce technique sensitivity as well as chair time. PMID- 17465662 TI - Bracket placement in lingual vs labial systems and direct vs indirect bonding. AB - OBJECTIVE: To examine the ultimate accuracy of bracket placement in labial vs lingual systems and in direct vs indirect bonding techniques. MATERIALS AND METHODS: Forty pretreatment dental casts of 20 subjects were selected. For each dental cast, four types of bracket placement were compared: labial direct (LbD), labial indirect (LbI), lingual direct (LgD), and lingual indirect (LgI). Direct bonding was performed with the casts held in a mannequin head. Labial brackets were oriented with a Boone gauge, and lingual brackets were oriented with the Lingual-Bracket-Jig System. Torque error (TqE) and rotation deviation (RotD) were measured with a torque geometric triangle and a toolmaker's microscope, respectively. Both torque and rotational measurements were evaluated statistically as algebraic and absolute numeric values, using analysis of variance with repeated measures. RESULTS: Absolute TqE and RotD were significantly (P < .001) higher in direct than in indirect bonding techniques higher in both the labial and lingual bracket systems by twofold and threefold, respectively (LbD = 7.26 degrees , 1.06 mm; vs LbI = 3.02 degrees , 0.75 mm; LgD = 8.42 degrees , 1.13 mm; vs LgI = 3.18 degrees , 0.55 mm). No statistically significant difference was found between labial and lingual systems for the same bonding technique. Maxillary incisors demonstrated the largest RotD angle (eg, right lateral: 12.04 degrees ). A distal off-center RotD was predominant in the mandibular dentition. CONCLUSIONS: Labial and lingual systems have the same level of inaccuracy. For both systems, indirect bonding significantly reduces absolute TqE and RotD. The TqE found can cause transverse discrepancy (scissors or crossbite) combined with disclusion with antagonist teeth. The RotD found can result in irregular interproximal contact points. PMID- 17465663 TI - Shear bond strength of orthodontic brackets bonded using conventional vs one and two step self-etching/adhesive systems. AB - OBJECTIVE: To assess and compare the effects of one- and two-step self-etching primer and adhesive with conventional acid-etching and bonding system on the shear bond strength of orthodontic brackets. MATERIALS AND METHODS: The one-step self-etching primer and adhesive used was Clearfil tri-S bond, the two-step fluoride-releasing antibacterial self-etching primer and adhesive was Clearfil Protect Bond, and the fluoride-releasing conventional acid-etching and bonding system was Kurasper F Bond. Brackets were bonded to defect-free human premolars (n = 14 per group) according to each manufacturer's recommendations by using light-cured bracket adhesive Kurasper F Paste with a light-emitting diode of a light-curing unit. The specimens were stored in deionized water at 37 degrees C for 48 hours and then tested in shear with a universal testing machine at a crosshead speed of 5 mm/min until the brackets debonded. The mode of failure of the brackets was determined by a modified adhesive remnant index. RESULTS: Mean shear bond strength values were 9.00 MPa for Kurasper F Bond, 9.55 MPa for Clearfil Protect Bond, and 9.48 MPa for Clearfil tri-S Bond. One-way analysis of variance detected no statistically significant difference among groups (P = .98, P > .05). The predominant failure for the three groups was at the bracket adhesive interface leaving less than 25% of the adhesive on the bracket base. CONCLUSIONS: One-step self-etching adhesive and two-step fluoride-releasing antibacterial self-etching adhesive have sufficient mechanical properties for the bonding of orthodontic brackets. PMID- 17465664 TI - Shear bond strength of precoated and uncoated brackets using a self-etching primer. AB - OBJECTIVE: To test the hypothesis that there are no significant differences in the shear bond strength or the adhesive remaining on the tooth after debonding between precoated and uncoated brackets using a self-etching primer. MATERIALS AND METHODS: APC Plus precoated brackets and uncoated brackets were bonded with Transbond XT adhesive using the self-etching primer Transbond Plus Self-Etching Primer (TPSEP). The brackets were bonded to extracted human premolars and categorized into two groups: (1) TPSEP/Transbond XT and (2) TPSEP/APC Plus. Shear bond strength was measured with a universal testing machine. The adhesive remnant on each tooth after debonding was quantified with image analysis equipment. Scanning electron microscope (SEM) observations of enamel surfaces treated with TPSEP were also carried out. RESULTS: No significant differences were observed in the shear bond strengths for the two groups evaluated (P < .05). TPSEP/APC Plus left significantly less adhesive on the tooth after debonding than did TPSEP/Transbond XT (P < .05). It was observed that SEM left a porous and potentially retentive surface. CONCLUSIONS: There was no significant difference in the bond strength of the two systems tested, but there was a significant difference in the percentage of area of adhesive remaining on the tooth. PMID- 17465665 TI - Surface roughness of acrylic resins after different curing and polishing techniques. AB - OBJECTIVE: To conduct a laboratory evaluation of the surface roughness of self curing acrylic resin after different curing and polishing techniques. MATERIALS AND METHODS: Sixty specimens were separated into four groups. The conventional curing process was followed by a second curing cycle in a microwave oven to decrease the residual monomer levels in two groups. After curing, two groups received manual polishing and the other groups underwent chemical polishing at 70 degrees C for 10 seconds. Roughness analysis was performed in a mechanical and laser profilometer. Analyses by Bartlett, Shapiro-Wilk, and Dunnett tests were used to compare whether the mean of the response variable was the same in all groups. RESULTS: The results suggested greater influences from the polishing method than the curing method and showed that the chemical polishing method yielded the highest surface roughness. CONCLUSIONS: The average pattern of roughness of the self-curing acrylic resin was statistically the same in the groups with different curing methods. However, chemical polishing increases the average pattern of roughness. PMID- 17465666 TI - Antibacterial activity and shear bond strength of 4-methacryloxyethyl trimellitate anhydride/methyl methacrylate-tri-n-butyl borane resin containing an antibacterial agent. AB - OBJECTIVE: To produce an antibacterial adhesive for orthodontic bonding without compromising the mechanical property. MATERIALS AND METHODS: We added benzalkonium chloride (BAC) to the Superbond C&B (4-methacryloxyethyl trimellitate anhydride/methyl methacrylate-tri-n-butyl borane [4-META/MMA-TBB]), a resin that exhibits a strong bonding strength between enamel and bracket. BAC concentrations in the BAC composites were 0.25%, 0.75%, 1.25%, 1.75%, 2.5%, and 5% (wt/wt). Antibacterial activity of the BAC composite was measured by the disk diffusion method. BAC-composite discs were placed on the surface of the agar inoculated with Streptococcus mutans and Streptococcus sobrinus, and the plates were incubated at 37 degrees C. After 48 hours of incubation, the inhibition zone around each sample was measured and recorded. The BAC-modified composite was used to bond metal brackets to the phosphoric acid-etched enamel surface of human premolars. The shear bond strengths were measured after immersion in water at 37 degrees C for 24 hours. RESULTS: The BAC-composite samples showed significant (P < .0001) antibacterial activity compared with the control. Measurable zones of bacterial inhibition increased as the BAC content in test samples increased. The shear bond strength declined with the increase in BAC concentration in the composite. A significant difference was found between the control composite and composites containing 1.25%, 1.75%, 2.5%, and 5% BAC (P < .05). No significant difference was found between the control composite and composites containing 0.25% and 0.75% BAC. However, shear bond strengths of the modified composites ranged from 10.12 MPa to 20.94 MPa. CONCLUSIONS: These results confirmed that BAC modified 4-META/MMA-TBB resin has a possibility for clinical application as an orthodontic bonding adhesive. PMID- 17465667 TI - Shear bond strength differences of types of maxillary deciduous and permanent teeth used as anchor teeth. AB - OBJECTIVE: To compare the bond strengths of identical orthodontic brackets bonded to maxillary deciduous and permanent teeth, which were used as anchor teeth in sectional edgewise appliance therapy, and to evaluate the modes of bracket/adhesive failure. MATERIALS AND METHODS: A total of 28 human maxillary teeth were divided into four groups by tooth type: permanent first premolars (group I), permanent second premolars (group II), deciduous canines (group III), and deciduous second molars (group IV). Each group consisted of seven teeth. Metal premolar brackets were bonded to these specimens using an acid-etching adhesive system. Shear bond strengths were measured using a universal testing machine, and the bracket/ adhesive failure modes were evaluated with the Adhesive Remnant Index. RESULTS: The shear strengths of the bonds on the deciduous canines and second molars were significantly lower than those on the permanent first and/or second premolars. There were no significant differences in shear bond strength between the permanent first and second premolars, and between the deciduous canines and second molars. The shear bond strengths of all four tooth types were higher than the clinically sufficient range of strengths from 6 to 8 MPa. Bond failure at the enamel-adhesive interface occurred more frequently in the deciduous second molars than in the permanent first premolars. CONCLUSIONS: There were significant differences in shear bond strength among different permanent and deciduous tooth types, but deciduous teeth could be used as anchor teeth in orthodontic treatment with sectional edgewise appliances. PMID- 17465668 TI - TMD in relation to malocclusion and orthodontic treatment. AB - OBJECTIVE: The aim of this systematic literature review was to evaluate associations between different malocclusions, orthodontic treatment, and signs and symptoms of temporomandibular disorders (TMD). MATERIALS AND METHODS: This review was part of a project at the Swedish Council on Technology Assessment in Health Care focusing on malocclusion and orthodontic treatment from a health perspective. As a first step, the literature was searched in the Medline and Cochrane Library databases from 1966 to May 2003. A later update was made in January 2005. Human studies in English or in Scandinavian languages were included. RESULTS: Associations between certain malocclusions and TMD were found in some studies, whereas the majority of the reviewed articles failed to identify significant and clinically important associations. TMD could not be correlated to any specific type of malocclusion, and there was no support for the belief that orthodontic treatment may cause TMD. Obvious individual variations in signs and symptoms of TMD over time according to some longitudinal studies further emphasized the difficulty in establishing malocclusion as a significant risk factor for TMD. A considerable reduction in signs and symptoms of TMD between the teenage period and young adulthood has been shown in some recent longitudinal studies. CONCLUSIONS: Associations between specific types of malocclusions and development of significant signs and symptoms of TMD could not be verified. There is still a need for longitudinal studies. PMID- 17465669 TI - Early correction of a developing skeletal Class III malocclusion. AB - This case report describes the treatment of a Japanese girl aged 11 years 10 months who had a severe Class III malocclusion with a concave facial profile. She presented hypodivergent skeletal pattern with a -4.0-mm anterior crossbite and a deep overbite. She also had facial asymmetry attributed partly to the lateral mandibular shift to avoid incisal interferences. The treatment plan included a monoblock appliance, rapid palatal expansion, and fixed edgewise appliances at the final stage. The monoblock appliance was used to redirect the growth of the mandible to a clockwise direction and simultaneously correct the incisal relationships along with fixed edgewise appliances. Good incisal relationships were achieved, and facial esthetics was greatly improved after 28 months of treatment. Stability of the treatment result was excellent in the 3-year 9-month follow-up at the age of 18. PMID- 17465670 TI - Orthodontic and surgical treatment of hemimandibular hyperplasia. AB - Correction of facial asymmetries is still a major problem in need of an adequate solution. In this case report, the differential diagnosis and treatment strategies of hemimandibular hyperplasia are described with the present patient. PMID- 17465671 TI - Orthodonticinformation.com. PMID- 17465672 TI - Deciphering protein-protein interactions. Part II. Computational methods to predict protein and domain interaction partners. AB - Recent advances in high-throughput experimental methods for the identification of protein interactions have resulted in a large amount of diverse data that are somewhat incomplete and contradictory. As valuable as they are, such experimental approaches studying protein interactomes have certain limitations that can be complemented by the computational methods for predicting protein interactions. In this review we describe different approaches to predict protein interaction partners as well as highlight recent achievements in the prediction of specific domains mediating protein-protein interactions. We discuss the applicability of computational methods to different types of prediction problems and point out limitations common to all of them. PMID- 17465673 TI - Dog as an outgroup to human and mouse. PMID- 17465674 TI - Coping with viral diversity in HIV vaccine design. AB - The ability of human immunodeficiency virus type 1 (HIV-1) to develop high levels of genetic diversity, and thereby acquire mutations to escape immune pressures, contributes to the difficulties in producing a vaccine. Possibly no single HIV-1 sequence can induce sufficiently broad immunity to protect against a wide variety of infectious strains, or block mutational escape pathways available to the virus after infection. The authors describe the generation of HIV-1 immunogens that minimizes the phylogenetic distance of viral strains throughout the known viral population (the center of tree [COT]) and then extend the COT immunogen by addition of a composite sequence that includes high-frequency variable sites preserved in their native contexts. The resulting COT(+) antigens compress the variation found in many independent HIV-1 isolates into lengths suitable for vaccine immunogens. It is possible to capture 62% of the variation found in the Nef protein and 82% of the variation in the Gag protein into immunogens of three gene lengths. The authors put forward immunogen designs that maximize representation of the diverse antigenic features present in a spectrum of HIV-1 strains. These immunogens should elicit immune responses against high-frequency viral strains as well as against most mutant forms of the virus. PMID- 17465675 TI - Patterns of mesenchymal condensation in a multiscale, discrete stochastic model. AB - Cells of the embryonic vertebrate limb in high-density culture undergo chondrogenic pattern formation, which results in the production of regularly spaced "islands" of cartilage similar to the cartilage primordia of the developing limb skeleton. The first step in this process, in vitro and in vivo, is the generation of "cell condensations," in which the precartilage cells become more tightly packed at the sites at which cartilage will form. In this paper we describe a discrete, stochastic model for the behavior of limb bud precartilage mesenchymal cells in vitro. The model uses a biologically motivated reaction diffusion process and cell-matrix adhesion (haptotaxis) as the bases of chondrogenic pattern formation, whereby the biochemically distinct condensing cells, as well as the size, number, and arrangement of the multicellular condensations, are generated in a self-organizing fashion. Improving on an earlier lattice-gas representation of the same process, it is multiscale (i.e., cell and molecular dynamics occur on distinct scales), and the cells are represented as spatially extended objects that can change their shape. The authors calibrate the model using experimental data and study sensitivity to changes in key parameters. The simulations have disclosed two distinct dynamic regimes for pattern self-organization involving transient or stationary inductive patterns of morphogens. The authors discuss these modes of pattern formation in relation to available experimental evidence for the in vitro system, as well as their implications for understanding limb skeletal patterning during embryonic development. PMID- 17465676 TI - Fundamental limits to position determination by concentration gradients. AB - Position determination in biological systems is often achieved through protein concentration gradients. Measuring the local concentration of such a protein with a spatially varying distribution allows the measurement of position within the system. For these systems to work effectively, position determination must be robust to noise. Here, we calculate fundamental limits to the precision of position determination by concentration gradients due to unavoidable biochemical noise perturbing the gradients. We focus on gradient proteins with first-order reaction kinetics. Systems of this type have been experimentally characterised in both developmental and cell biology settings. For a single gradient we show that, through time-averaging, great precision potentially can be achieved even with very low protein copy numbers. As a second example, we investigate the ability of a system with oppositely directed gradients to find its centre. With this mechanism, positional precision close to the centre improves more slowly with increasing averaging time, and so longer averaging times or higher copy numbers are required for high precision. For both single and double gradients, we demonstrate the existence of optimal length scales for the gradients for which precision is maximized, as well as analyze how precision depends on the size of the concentration-measuring apparatus. These results provide fundamental constraints on the positional precision supplied by concentration gradients in various contexts, including both in developmental biology and also within a single cell. PMID- 17465677 TI - RNAi development. PMID- 17465678 TI - Identification of a CCR5-expressing T cell subset that is resistant to R5-tropic HIV infection. AB - Infection with HIV-1 perturbs homeostasis of human T cell subsets, leading to accelerated immunologic deterioration. While studying changes in CD4(+) memory and naive T cells during HIV-1 infection, we found that a subset of CD4(+) effector memory T cells that are CCR7(-)CD45RO(-)CD45RA(+) (referred to as TEMRA cells), was significantly increased in some HIV-infected individuals. This T cell subset displayed a differentiated phenotype and skewed Th1-type cytokine production. Despite expressing high levels of CCR5, TEMRA cells were strikingly resistant to infection with CCR5 (R5)-tropic HIV-1, but remained highly susceptible to CXCR4 (X4)-tropic HIV-1. The resistance of TEMRA cells to R5 tropic viruses was determined to be post-entry of the virus and prior to early viral reverse transcription, suggesting a block at the uncoating stage. Remarkably, in a subset of the HIV-infected individuals, the relatively high proportion of TEMRA cells within effector T cells strongly correlated with higher CD4(+) T cell numbers. These data provide compelling evidence for selection of an HIV-1-resistant CD4(+) T cell population during the course of HIV-1 infection. Determining the host factors within TEMRA cells that restrict R5-tropic viruses and endow HIV-1-specific CD4(+) T cells with this ability may result in novel therapeutic strategies against HIV-1 infection. PMID- 17465679 TI - A novel TLR4-mediated signaling pathway leading to IL-6 responses in human bladder epithelial cells. AB - The vigorous cytokine response of immune cells to Gram-negative bacteria is primarily mediated by a recognition molecule, Toll-like receptor 4 (TLR4), which recognizes lipopolysaccharide (LPS) and initiates a series of intracellular NF kappaB-associated signaling events. Recently, bladder epithelial cells (BECs) were reported to express TLR4 and to evoke a vigorous cytokine response upon exposure to LPS. We examined intracellular signaling events in human BECs leading to the production of IL-6, a major urinary cytokine, following activation by Escherichia coli and isolated LPS. We observed that in addition to the classical NF-kappaB-associated pathway, TLR4 triggers a distinct and more rapid signaling response involving, sequentially, Ca(2+), adenylyl cyclase 3-generated cAMP, and a transcriptional factor, cAMP response element-binding protein. This capacity of BECs to mobilize secondary messengers and evoke a more rapid IL-6 response might be critical in their role as first responders to microbial challenge in the urinary tract. PMID- 17465680 TI - Dietary restriction in Drosophila: delayed aging or experimental artefact? AB - Lifespan can be extended by reduction of dietary intake. This practice is referred to as dietary restriction (DR), and extension of lifespan by DR is evolutionarily conserved in taxonomically diverse organisms including yeast, invertebrates, and mammals. Although these two often-stated facts carry the implication that the mechanisms of DR are also evolutionarily conserved, extension of lifespan could be a case of evolutionary convergence, with different underlying mechanisms in different taxa. Furthermore, extension of lifespan by different methods of DR in the same organism may operate through different mechanisms. These topics remain unresolved because of the very fact that the mechanisms of DR are unknown. Given these uncertainties, it is essential that work on the mechanisms of DR is not clouded by imprecise descriptions of methods or by technical problems. Here we review the recent literature on DR in Drosophila to point out some methodological issues that can obscure mechanistic interpretations. We also indicate some experiments that could be performed to determine if DR in Drosophila operates through similar mechanisms to the process in rodents. PMID- 17465681 TI - The association of a SNP upstream of INSIG2 with body mass index is reproduced in several but not all cohorts. AB - A SNP upstream of the INSIG2 gene, rs7566605, was recently found to be associated with obesity as measured by body mass index (BMI) by Herbert and colleagues. The association between increased BMI and homozygosity for the minor allele was first observed in data from a genome-wide association scan of 86,604 SNPs in 923 related individuals from the Framingham Heart Study offspring cohort. The association was reproduced in four additional cohorts, but was not seen in a fifth cohort. To further assess the general reproducibility of this association, we genotyped rs7566605 in nine large cohorts from eight populations across multiple ethnicities (total n = 16,969). We tested this variant for association with BMI in each sample under a recessive model using family-based, population based, and case-control designs. We observed a significant (p < 0.05) association in five cohorts but saw no association in three other cohorts. There was variability in the strength of association evidence across examination cycles in longitudinal data from unrelated individuals in the Framingham Heart Study Offspring cohort. A combined analysis revealed significant independent validation of this association in both unrelated (p = 0.046) and family-based (p = 0.004) samples. The estimated risk conferred by this allele is small, and could easily be masked by small sample size, population stratification, or other confounders. These validation studies suggest that the original association is less likely to be spurious, but the failure to observe an association in every data set suggests that the effect of SNP rs7566605 on BMI may be heterogeneous across population samples. PMID- 17465683 TI - Mitotic recombination accelerates adaptation in the fungus Aspergillus nidulans. AB - Understanding the prevalence of sexual reproduction in eukaryotes is a hard problem. At least two aspects still defy a fully satisfactory explanation, the functional significance of genetic recombination and the great variation among taxa in the relative lengths of the haploid and diploid phases in the sexual cycle. We have performed an experimental study to explore the specific advantages of haploidy or diploidy in the fungus Aspergillus nidulans. Comparing the rate of adaptation to a novel environment between haploid and isogenic diploid strains over 3,000 mitotic generations, we demonstrate that diploid strains, which during the experiment have reverted to haploidy following parasexual recombination, reach the highest fitness. This is due to the accumulation of recessive deleterious mutations in diploid nuclei, some of which show their combined beneficial effect in haploid recombinants. Our findings show the adaptive significance of mitotic recombination combined with flexibility in the timing of ploidy level transition if sign epistasis is an important determinant of fitness. PMID- 17465682 TI - PPAR gamma 2 prevents lipotoxicity by controlling adipose tissue expandability and peripheral lipid metabolism. AB - Peroxisome proliferator activated receptor gamma 2 (PPARg2) is the nutritionally regulated isoform of PPARg. Ablation of PPARg2 in the ob/ob background, PPARg2(-/ ) Lep(ob)/Lep(ob) (POKO mouse), resulted in decreased fat mass, severe insulin resistance, beta-cell failure, and dyslipidaemia. Our results indicate that the PPARg2 isoform plays an important role, mediating adipose tissue expansion in response to positive energy balance. Lipidomic analyses suggest that PPARg2 plays an important antilipotoxic role when induced ectopically in liver and muscle by facilitating deposition of fat as relatively harmless triacylglycerol species and thus preventing accumulation of reactive lipid species. Our data also indicate that PPARg2 may be required for the beta-cell hypertrophic adaptive response to insulin resistance. In summary, the PPARg2 isoform prevents lipotoxicity by (a) promoting adipose tissue expansion, (b) increasing the lipid-buffering capacity of peripheral organs, and (c) facilitating the adaptive proliferative response of beta-cells to insulin resistance. PMID- 17465684 TI - The tailless ortholog nhr-67 regulates patterning of gene expression and morphogenesis in the C. elegans vulva. AB - Regulation of spatio-temporal gene expression in diverse cell and tissue types is a critical aspect of development. Progression through Caenorhabditis elegans vulval development leads to the generation of seven distinct vulval cell types (vulA, vulB1, vulB2, vulC, vulD, vulE, and vulF), each with its own unique gene expression profile. The mechanisms that establish the precise spatial patterning of these mature cell types are largely unknown. Dissection of the gene regulatory networks involved in vulval patterning and differentiation would help us understand how cells generate a spatially defined pattern of cell fates during organogenesis. We disrupted the activity of 508 transcription factors via RNAi and assayed the expression of ceh-2, a marker for vulB fate during the L4 stage. From this screen, we identified the tailless ortholog nhr-67 as a novel regulator of gene expression in multiple vulval cell types. We find that one way in which nhr-67 maintains cell identity is by restricting inappropriate cell fusion events in specific vulval cells, namely vulE and vulF. nhr-67 exhibits a dynamic expression pattern in the vulval cells and interacts with three other transcriptional regulators cog-1 (Nkx6.1/6.2), lin-11 (LIM), and egl-38 (Pax2/5/8) to generate the composite expression patterns of their downstream targets. We provide evidence that egl-38 regulates gene expression in vulB1, vulC, vulD, vulE, as well as vulF cells. We demonstrate that the pairwise interactions between these regulatory genes are complex and vary among the seven cell types. We also discovered a striking regulatory circuit that affects a subset of the vulval lineages: cog-1 and nhr-67 inhibit both one another and themselves. We postulate that the differential levels and combinatorial patterns of lin-11, cog-1, and nhr-67 expression are a part of a regulatory code for the mature vulval cell types. PMID- 17465685 TI - Genetic basis for dosage sensitivity in Arabidopsis thaliana. AB - Aneuploidy, the relative excess or deficiency of specific chromosome types, results in gene dosage imbalance. Plants can produce viable and fertile aneuploid individuals, while most animal aneuploids are inviable or developmentally abnormal. The swarms of aneuploid progeny produced by Arabidopsis triploids constitute an excellent model to investigate the mechanisms governing dosage sensitivity and aneuploid syndromes. Indeed, genotype alters the frequency of aneuploid types within these swarms. Recombinant inbred lines that were derived from a triploid hybrid segregated into diploid and tetraploid individuals. In these recombinant inbred lines, a single locus, which we call SENSITIVE TO DOSAGE IMBALANCE (SDI), exhibited segregation distortion in the tetraploid subpopulation only. Recent progress in quantitative genotyping now allows molecular karyotyping and genetic analysis of aneuploid populations. In this study, we investigated the causes of the ploidy-specific distortion at SDI. Allele frequency was distorted in the aneuploid swarms produced by the triploid hybrid. We developed a simple quantitative measure for aneuploidy lethality and using this measure demonstrated that distortion was greatest in the aneuploids facing the strongest viability selection. When triploids were crossed to euploids, the progeny, which lack severe aneuploids, exhibited no distortion at SDI. Genetic characterization of SDI in the aneuploid swarm identified a mechanism governing aneuploid survival, perhaps by buffering the effects of dosage imbalance. As such, SDI could increase the likelihood of retaining genomic rearrangements such as segmental duplications. Additionally, in species where triploids are fertile, aneuploid survival would facilitate gene flow between diploid and tetraploid populations via a triploid bridge and prevent polyploid speciation. Our results demonstrate that positional cloning of loci affecting traits in populations containing ploidy and chromosome number variants is now feasible using quantitative genotyping approaches. PMID- 17465686 TI - Training emergency medicine doctors for rural and regional Australia: can we learn from other countries? AB - INTRODUCTION: Australia is a country with a relatively small rural population dispersed over an enormous area. Issues similar to how best to deliver health services and recruit health professionals to rural areas exist in other countries. For professional and lifestyle reasons, most specialist doctors (including emergency medicine specialists), choose to live and work in major metropolitan centres. Outside the major Australian cities, most presentations to emergency departments are dealt with by 'non-specialist' doctors, often with limited specialist back up. Recruitment of suitably trained medical staff is increasingly difficult. There is increasing reliance on overseas trained doctors from widely varying backgrounds. In Canada and New Zealand, family medicine trained emergency medicine doctors are a significant proportion of the workforce in rural and regional emergency departments. AIM: To undertake a detailed investigation of the non-specialist emergency medicine doctors in Australia, and examine strategies to secure a more highly trained emergency medicine workforce for rural and regional Australia. METHODS: A survey was undertaken of 230 doctors working in 57 rural and regional Australian emergency departments. Consultations and interviews were held with 53 key clinicians, educators and administrators. RESULTS: There were no training or education standards for doctors working in smaller Australian emergency departments. There was considerable instability in the workforce with many planning to leave their current position or reduce their involvement in emergency medicine. The vast majority felt a need to undertake further emergency medicine training. There was little agreement among stakeholders as to how emergency medicine should be taught, or services delivered in rural and regional Australia. CONCLUSIONS: There is need in Australia to offer a specific postgraduate qualification in emergency medicine for doctors wishing to practise emergency medicine outside major city hospitals. Ideally, such a course would be largely taught in rural and regional hospitals and would contain additional elements relevant to rural practice. The Canadian and New Zealand emergency medicine qualifications may be useful models. PMID- 17465687 TI - Bringing rural nurses, midwives & rural communities closer together: eliminating the isolation barrier through technology. PMID- 17465688 TI - Effect of serotonin on small intestinal contractility in healthy volunteers. AB - The physiological significance of serotonin released into the intestinal lumen for the regulation of motility is unknown in humans. The aim of this study was to evaluate the effect of serotonin infused into the lumen of the gastric antrum, duodenum or the jejunum, on antro-duodeno-jejunal contractility in healthy human volunteers. Manometric recordings were obtained and the effects of either a standard meal, continuous intravenous infusion of serotonin (20 nmol/kg/min) or intraluminal bolus infusions of graded doses of serotonin (2.5, 25 or 250 nmol) were compared. In addition, platelet-depleted plasma levels of serotonin, blood pressure, heart rate and electrocardiogram were evaluated. All subjects showed similar results. Intravenous serotonin increased migrating motor complex phase III frequency 3-fold and migrating velocity 2-fold. Intraluminal infusion of serotonin did not change contractile activity. Platelet-depleted-plasma levels of serotonin increased 2-fold following both intravenous and high doses of intraluminal infusions of serotonin. All subjects reported minor short-lived adverse effects following intravenous serotonin stimulation, while only half of the subjects reported minor short-lived adverse effects following intraluminal serotonin stimulations. We conclude that exogenous serotonin in the lumen of the upper part of the small intestine does not seem to change antro-duodeno-jejunal contractility significantly in healthy adult volunteers. PMID- 17465689 TI - Ryanodine receptors, voltage-gated calcium channels and their relationship with protein kinase A in the myocardium. AB - We present a review about the relationship between ryanodine receptors and voltage-gated calcium channels in myocardium, and also how both of them are related to protein kinase A. Ryanodine receptors, which have three subtypes (RyR1 3), are located on the membrane of sarcoplasmic reticulum. Different subtypes of voltage-gated calcium channels interact with ryanodine receptors in skeletal and cardiac muscle tissue. The mechanism of excitation-contraction coupling is therefore different in the skeletal and cardiac muscle. However, in both tissues ryanodine receptors and voltage-gated calcium channels seem to be physically connected. FK-506 binding proteins (FKBPs) are bound to ryanodine receptors, thus allowing their concerted activity, called coupled gating. The activity of both ryanodine receptors and voltage-gated calcium channels is positively regulated by protein kinase A. These effects are, therefore, components of the mechanism of sympathetic stimulation of myocytes. The specificity of this enzyme's targeting is achieved by using different A kinase adapting proteins. Different diseases are related to inborn or acquired changes in ryanodine receptor activity in cardiac myocytes. Mutations in the cardiac ryanodine receptor gene can cause catecholamine-provoked ventricular tachycardia. Changes in phosphorylation state of ryanodine receptors can provide a credible explanation for the development of heart failure. The restoration of their normal level of phosphorylation could explain the positive effect of beta-blockers in the treatment of this disease. In conclusion, molecular interactions of ryanodine receptors and voltage-gated calcium channels with PKA have a significant physiological role. However, their defects and alterations can result in serious disturbances. PMID- 17465690 TI - Effect of chronic cadmium exposure on antioxidant defense system in some tissues of rats: protective effect of selenium. AB - The effects of selenium (Se) on antioxidant defense system in liver and kidneys of rats with cadmium (Cd)-induced toxicity were examined. Cd exposure (15 mg Cd/kg b.m./day as CdCl(2) for 4 weeks) resulted in increased lipid peroxidation (LP) in both organs (p<0.005 and p<0.01). Vitamin C (Vit C) was decreased in the liver (p<0.005), whereas vitamin E (Vit E) was increased in the liver and kidneys (p<0.005 and p<0.05) of Cd-exposed animals. Superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) activities were decreased in both tissues (p<0.05 and p<0.005), whereas catalase (CAT) activity was decreased only in liver (p<0.005). Glutathione S-transferase (GST) increased in both tissues (p<0.005 and p<0.01). Treatment with Se (0.5 mg Se/kg b.m./day as Na(2)SeO(3) for 4 weeks) significantly increased liver and kidneys SOD and GSH-Px activities (p<0.05 to p<0.005), as well as CAT and GST activities only in the liver (p<0.01). In animals exposed to Se, both the concentrations of Vit C (p<0.01) and Vit E (p<0.005) were increased in both tissues. Co-treatment with Se resulted in reversal of oxidative stress with significant decline in analyzed tissues Cd burden. Our results show that Se may ameliorate Cd-induced oxidative stress by decreasing LP and altering antioxidant defense system in rat liver and kidneys and that Se demonstrates the protective effect from cadmium-induced oxidative damage. PMID- 17465691 TI - Effects of endothelin-1 1-31 on cell viabililty and [Ca2+]i in cultured neonatal rat cardiomyocytes. AB - We previously found that endothelin-1(1-31) (ET-1(1-31)) exhibited a pro arrhythmogenic effect in isolated rat hearts. In this study, we further investigated the effects of ET-1(1-31) on a cell viability and observed [Ca(2+)](i) in cultured cardiomyocytes. Cultured neonatal rat cardiomyocytes were treated with 0.1, 1, and 10 nM ET-1(1-31) for 24h in the presence or absence of ET(A) receptor antagonist (BQ(123)) or phosphoramidon, a NEP/ECE inhibitor. Cell injury was evaluated by supernatant lactate dehydrogenase (LDH) assay, superoxide dismutase (SOD) activity, and malondialdehyde (MDA) content. Cell viability was assessed by MTT assay. [Ca(2+)](i) was measured with Fluo-3/AM under a laser confocal microscope. 1) ET-1(1-31) dose-dependently increased LDH release and decreased cell viability. 2) LDH and MDA levels were significantly elevated and SOD activity decreased after administration of 1 nM ET-1(1-31) for 24h, and these changes were markedly attenuated by 1 uM BQ(123). 3) Exposure to 10 nM ET 1(1-31) caused a continuous increase in [Ca(2+)](i) to cultured beating cardiomyocytes and termination of [Ca(2+)](i) transient within 6 min, and this change was reversed by 1 uM BQ(123) and attenuated by 0.5 mM phosphoramidon. These results suggest that ET-1(1-31) could cause cell injury, and that the effect of ET-1(1 31) on [Ca(2+)](i) transients is mainly mediated by ET(A) receptor and partially attributed to the conversion of ET-1(1-31) to ET-1(1-21). PMID- 17465692 TI - Bone remodeling, particle disease and individual susceptibility to periprosthetic osteolysis. AB - Bone remodeling is a tightly coupled process consisting of repetitive cycles of bone resorption and formation. Both processes are governed by mechanical signals, which operate in conjunction with local and systemic factors in a discrete anatomic structure designated a basic multicellular unit (BMU). The microenvironment around total joint arthroplasty is a dynamic and complex milieu influenced by the chemical and physical stimuli associated with servicing the prosthesis. A key factor limiting the longevity of the prosthesis is polyethylene wear, which induces particle disease, and this may lead to increased and prolonged activity of BMUs resulting in periprosthetic osteolysis. Several pathways regulating BMU function have been reported in the past, including RANKL/RANK/OPG/TRAF6, TNF-alpha/TNFR/TRAF1, and IL-6/CD126/JAK/STAT. Moreover, the expression and functional activity of all these molecules can be affected by variations in their genes. These may explain the differences in severity of bone defects or prosthetic failure between patients with similar wear rates and the same prosthesis. Simultaneously, this data strongly support the theory of individual susceptibility to prosthetic failure. PMID- 17465693 TI - Leptin affects proliferation-, apoptosis- and protein kinase A-related peptides in human ovarian granulosa cells. AB - The aim of our in vitro studies was to understand the role of leptin in controlling proliferation, apoptosis, and protein kinase A (PKA) in human ovarian cells. We analyzed the in vitro effects of leptin (0, 1, 10 or 100 ng/ml) on the accumulation of proliferation-related peptides (PCNA, cyclin B1), apoptosis associated peptide (Bax) and the intracellular signaling molecule PKA in cultured human granulosa cells using immunocytochemistry and Western immunoblotting. It was observed that leptin stimulated in a dose-dependent manner the accumulation of PCNA (at doses 1-100 ng/ml), cyclin B1 (at doses 10 or 100 ng/ml), Bax (at doses 10 or 100 ng/ml) and PKA (at doses 1-100 ng/ml) in cultured human ovarian cells. These observations suggest the ability of leptin to control directly human ovarian cell functions: proliferation, apoptosis, and intracellular messenger PKA. PMID- 17465694 TI - Catheter-based management of aortic valve regurgitation in experimental cardiology. AB - Non-surgical management of aortic valve disease has been given considerable attention. Several recent publications have already reported its use in clinical practice. The main issue is to get an understanding of the pathophysiological processes and, most importantly, extensive experimental activity. In addition to testing various animal models, technical and material aspects are also being intensively investigated. It is not clear yet whether the durability and applicability of this promising development will be comparable with the standard of current cardiac surgery. Nonetheless, even the use of some models as a temporary approach helping to improve the circulatory status, not allowing safe surgery, is certainly justified. At any rate, a new stage of research and clinical application has been set off. However, experimental background continues to be simply indispensable. The paper is a short review of the issue. PMID- 17465695 TI - The influence of wine polyphenols on reactive oxygen and nitrogen species production by murine macrophages RAW 264.7. AB - The aim was to study the antioxidant properties of four wine polyphenols (flavonoids catechin, epicatechin, and quercetin, and hydroxystilbene resveratrol). All three flavonoids exerted significant and dose-dependent scavenging effects against peroxyl radical and nitric oxide in chemical systems. The scavenging effect of resveratrol was significantly lower. All polyphenols decreased production of reactive oxygen species (ROS) by RAW264.7 macrophages. Only quercetin quenched ROS produced by lipopolysaccharide-stimulated RAW264.7 macrophages incubated for 24 h with polyphenols. Quercetin and resveratrol decreased the release of nitric oxide by these cells in a dose-dependent manner which corresponded to a decrease in iNOS expression in the case of quercetin. In conclusion, the higher number of hydroxyl substituents is an important structural feature of flavonoids in respect to their scavenging activity against ROS and nitric oxide, while C-2,3 double bond (present in quercetin and resveratrol) might be important for inhibition of ROS and nitric oxide production by RAW 264.7 macrophages. PMID- 17465696 TI - Neurobiological aspects of depressive disorder and antidepressant treatment: role of glia. AB - Depression is a complex disorder related to chronic inflammatory processes, chronic stress changes and a hippocampal response. There is a increasing knowledge about the role of glial cells in nutrient supply to neurons, maintenance of synaptic contacts and tissue homeostasis within the CNS. Glial cells, viewed in the past as passive elements with a limited influence on neuronal function, are becoming recognized as active partners of neurons and are starting to be discussed as a possible therapeutic target. Their role in the pathogenesis of depressive disorders is also being reconsidered. Attention is devoted to studies of the different types of antidepressants and their effects on transmembrane signaling, including levels of alpha subunits of G proteins in C6 glioma cells in vitro as a model of postsynaptic changes in vivo. These models indicate similarities in antidepressant effects on G proteins of brain cells and effector cells of natural immunity, natural killers and granulocytes. Thus, an antidepressant response can exhibit certain common characteristics in functionally different systems which also participate in disease pathogenesis. There are, however, differences in the astrocyte G-protein responses to antidepressant treatment, indicating that antidepressants differ in their effect on glial signalization. Today mainstream approach to neurobiological basis of depressive disorders and other mood illnesses is linked to abnormalities in transmembrane signal transduction via G-protein coupled receptors. Intracellular signalization cascade modulation results in the activation of transcription factors with subsequent increased production of a wide array of products including growth factors and to changes in cellular activity and reactivity. PMID- 17465697 TI - Neuroendocrine and oxidoreductive mechanisms of stress-induced cardiovascular diseases. AB - The review concerns a number of basic molecular pathways that play a crucial role in perception, transmission, and modulation of the stress signals, and mediate the adaptation of the vital processes in the cardiovascular system (CVS). These highly complex systems for intracellular transfer of information include stress hormones and their receptors, stress-activated phosphoprotein kinases, stress activated heat shock proteins, and antioxidant enzymes maintaining oxidoreductive homeostasis of the CVS. Failure to compensate for the deleterious effects of stress may result in the development of different pathophysiological states of the CVS, such as ischemia, hypertension, atherosclerosis and infarction. Stress induced dysbalance in each of the CVS molecular signaling systems and their contribution to the CVS malfunctioning is reviewed. The general picture of the molecular mechanisms of the stress-induced pathophysiology in the CVS pointed out the importance of stress duration and intensity as etiological factors, and suggested that future studies should be complemented by the careful insights into the individual factors of susceptibility to stress, prophylactic effects of 'healthy' life styles and beneficial action of antioxidant-rich nutrition. PMID- 17465698 TI - Intracerebroventricular injection of oxidant and antioxidant molecules affects long-term potentiation in urethane anaesthetized rats. AB - Production of superoxide anions in the incubation medium of hippocampal slices can induce long-term potentiation (LTP). Other reactive oxygen species (ROS) such as hydrogen peroxide are able to modulate LTP and are likely to be involved in aging mechanisms. The present study explored whether intracerebro-ventricular (ICV) injection of oxidant or antioxidant molecules could affect LTP in vivo. With this aim in mind, field excitatory post-synaptic potentials (fEPSPs) elicited by stimulation of the perforant pathway were recorded in the dentate gyrus of the hippocampal formation in urethane-anesthetized rats. N-acetyl-L cysteine, hydrogen peroxide (H2O2) or hypoxanthine/xanthine-oxidase solution (a superoxide producing system) were administrated by ICV injection. The control was represented by a group injected with saline ICV. Ten minutes after the injection, LTP was induced in the granule cells of the dentate gyrus by high frequency stimulation of the perforant pathway. Neither the H(2)O(2) injection or the N acetyl-L-cysteine injection caused any variation in the fEPSP at the 10-min post injection time point, whereas the superoxide generating system caused a significant increase in the fEPSP. Moreover, at 60 min after tetanic stimulation, all treatments attenuated LTP compared with the control group. These results show that ICV administration of oxidant or antioxidant molecules can modulate LTP in vivo in the dentate gyrus. Particularly, a superoxide producing system can induce potentiation of the synaptic response. Interestingly, ICV injection of oxidants or antioxidants prevented a full expression of LTP compared to the saline injection. PMID- 17465699 TI - Inhibition of palmityl carnitine oxidation in rat liver mitochondria by tert butyl hydroperoxide. AB - Mitochondria as an energy generating cell device are very sensitive to oxidative damage. Our previous findings obtained in hepatocytes demonstrated that Complex I of the respiratory chain is more sensitive to oxidative damage than other respiratory chain complexes. We present additional data on isolated mitochondria showing that palmityl carnitine oxidation is strongly depressed at a low (200 microM) tert-butyl hydroperoxide (tBHP) concentration, while oxidation of the flavoprotein-dependent substrate-succinate is not affected and neither is ATP synthesis inhibited by tBHP. In the presence of tBHP, the respiratory control index for palmityl carnitine oxidation is strongly depressed, but when succinate is oxidized the respiratory control index remains unaffected. Our findings thus indicate that flavoprotein-dependent substrates could be an important nutritional factor for the regeneration process in the necrotic liver damaged by oxidative stress. PMID- 17465700 TI - Leukocyte and endothelial adhesion molecules in patients with hypercholesterolemia: the effect of atorvastatin treatment. AB - Atherogenesis involves the migration of leukocytes into vascular subendothelial space, a process mediated by endothelial and leukocyte cell adhesion molecules. Endothelial molecules are assessed indirectly via serum levels, but leukocyte molecules can be assessed directly. We have therefore hypothesized that leukocyte adhesion molecules are altered to a greater degree in hypercholesterolemia than serum endothelial adhesion molecules. We examined 29 subjects with hypercholesterolemia and 27 controls at baseline and after 12 weeks of atorvastatin treatment (20 mg/day). Expression of leukocyte integrins CD11a, CD11b, CD18, and CD49d and of L-selectin was measured by flow cytometry. Serum ICAM-1, E-selectin and von Willebrand factor were measured by ELISA. Expression of leukocyte adhesion molecules was significantly higher in patients at baseline than in the controls, except for CD11a. Expression significantly decreased after atorvastatin in most adhesion molecules except for CD11b. In contrast, there was no effect of hypercholesterolemia and/or atorvastatin on the serum endothelial molecules. Leukocyte but not endothelial adhesion molecules were influenced by hypercholesterolemia and by lipid lowering treatment. Leukocyte molecules may therefore be a more sensitive marker of atherogenesis than endothelial molecules. Our results support the role of increased leukocyte adhesiveness in atherogenesis. PMID- 17465701 TI - Noninvasive in vivo assessment of the skeletal muscle and small intestine serous surface microcirculation in rat: sidestream dark-field (SDF) imaging. AB - The pathophysiology of microcirculation is intensively investigated to understand disease development at the microscopic level. Orthogonal polarization spectral (OPS) imaging and its successor sidestream dark-field (SDF) imaging are relatively new noninvasive optical techniques allowing direct visualization of microcirculation in both clinical and experimental studies. The goal of this experimental study was to describe basic microcirculatory parameters of skeletal muscle and ileal serous surface microcirculation in the rat using SDF imaging and to standardize the technical aspects of the protocol. Interindividual variability in functional capillary density (FCD) and small vessels (<25 microm in diameter) proportion was determined in anesthetized rats on the surface of quadriceps femoris (m. rectus femoris and m. vastus medialis) and serous surface of ileum. Special custom made flexible arm was used to fix the SDF probe minimizing the pressure movement artifacts. Clear high contrast images were analyzed off-line. The mean FCD obtained from the surface of skeletal muscle and ileal serous surface was 219 (213-225 cm/cm(2)) and 290 (282-298 cm/cm(2)) respectively. There was no statistically significant difference between rats in mean values of FCD obtained from the muscle (P = 0.273) in contrast to ileal serous surface, where such difference was statistically significant (P = 0.036). No statistically significant differences in small vessels percentage was detected on either the muscle surface (P = 0.739) or on ileal serous surface (P = 0.659). Our study has shown that interindividual variability of basic microcirculatory parameters in rat skeletal muscle and ileum is acceptable when using SDF imaging technique according to a highly standardized protocol and with appropriate fixation device. SDF imaging represents promising technology for experimental and clinical studies. PMID- 17465702 TI - Morphological changes of rat jejunum after whole body gamma-irradiation and their impact in biodosimetry. AB - Gastrointestinal form is the second stage of the Acute Radiation Syndrome (ARS) with a threshold dose of 8 Gy. It represents an absolutely lethal clinical pathological unit, enteritis necro-hemorrhagica (duodenitis, jejunitis, ileitis, respectively) with unknown causal therapy. The purpose of our study has been to evaluate the morphological changes in a model of radiation-induced enteritis in rats and estimate the significance of changes in biodosimetry. Wistar rats were randomly divided into 21 groups, 10 animals per group. Samples of the jejunum were taken 24, 48, 72, and 96 h after the whole-body gamma-irradiation with the doses of 1, 5, 10, 15, and 20 Gy, and routinely stained with hematoxylin and eosin. Five morphometric markers--intercryptal distance, enterocytal height on the top and base of villus, length of basal lamina of 10 enterocytes and enterocytal width--in irradiated rat jejunum were examined. The results were compared with sham-irradiated control group. After lethal doses of irradiation, all morphometric parameters of jejunum significantly changed. With the exception of intercryptal distance, they might be considered as suitable biodosimetric markers under these experimental conditions. Our morphometry results in radiation induced jejunitis are in accordance with those in other studies. We were the first who quantified morphological post-irradiation changes in animal jejunum. Some of them might be used under experimental conditions. This experimental study is a predecessor of the clinical assessment of a specific marker. Under clinical practice, the sensitive biodosimetric parameter could serve as one of the guidance for evaluation of the absorbed dose in irradiated troops as well as rescue workers. This is in accordance with tasks and Standardization Agreement of the North Atlantic Treaty Organization. PMID- 17465703 TI - Adiponectin as a potential marker of prostate cancer progression: studies in organ-confined and locally advanced prostate cancer. AB - Serum levels of adiponectin were measured in patients with benign prostatic hyperplasia and prostate cancer of pT2 and pT3 stage. Adiponectin ELISA assay, immunohistochemistry, and selected metabolic and biochemical parameters measurement was performed in 25 patients with benign prostatic hyperplasia and 43 with prostate cancer (17 patients with organ-confined and 26 patients with locally advanced disease). Serum adiponectin levels did not differ between prostate benign hyperplasia and cancer clinical stage T2, but was significantly higher in pT3 relative to pT2 group (14.51+/-4.92 vs. 21.41+/-8.12, P = 0.003). Tissue immunohistochemistry showed enhanced staining in neoplastic prostate glands and intraepithelial neoplasia relative to benign prostatic hyperplasia without distinction between disease grade and stage. Serum adiponectin levels are higher in locally advanced relative to organ-confined prostate cancer and may thus serve as an auxiliary marker providing further improvement for discrimination between pT2 and pT3 stages. PMID- 17465704 TI - Nutrigenomics in perspective. PMID- 17465705 TI - Future of toxicogenomics and safety signatures: balancing public access to data with proprietary drug discovery. PMID- 17465706 TI - Legislation to prevent genetic discrimination approved. PMID- 17465707 TI - Glutathione S-transferase M1 null genotype as a risk factor for carbamazepine induced mild hepatotoxicity. AB - The aim of this study is to verify whether the combination of glutathione S transferase (GST) M1 null and GSTT1 null genotypes, which is a candidate genetic risk factor for troglitazone-induced liver failure, is common to that for the carbamazepine-induced mild hepatotoxicity. PATIENTS & METHODS: The genotypes of GSTM1 and GSTT1, and microsomal epoxide hydrolase-3 and -4, were determined in 192 Japanese epileptics treated with carbamazepine. RESULTS: The GSTM1 null (GSTM1-) and GSTT1 null (GSTT1-) genotypes in the subjects were 55.7 and 39.6%, respectively. The alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels were elevated in 46 (24.0%) and 62 (32.3%) cases, and the mean values were approximately 2.3- and 1.8-times higher than the upper limit of normal levels, respectively. The levels of ALT and AST were significantly higher in GSTM1- than in GSTM1 present (GSTM1+) genotypes (p = 0.007 and 0.004, respectively). The level of ALT was significantly higher in GSTM1-/T1- than in GSTM1+/T1- and GSTM1+/T1+ (p = 0.01 and 0.01, respectively), and that of AST was significantly higher in GSTM1-/T1- and GSTM1-/T1+ than in GSTM1+/T1+ (p = 0.02 and 0.003, respectively). The microsomal epoxide hydrolase genotype did not influence the hepatotoxicity. CONCLUSION: These findings suggested that GSTM1- rather than GSTM1-/T1- was a risk factor for carbamazepine-induced mild hepatotoxicity. PMID- 17465708 TI - Sex-dependent genetic markers of CYP3A4 expression and activity in human liver microsomes. AB - OBJECTIVE: To find genetic markers of the individual cytochrome P450 (CYP)3A expression. METHODS: A large collection of liver samples phenotyped for CYP3A expression and activity was genotyped for CYP3A variants. Data were analyzed for associations between CYP3A phenotypes and genotypes, and for evidence of recent selection. RESULTS: We report associations between the hepatic CYP3A4 protein expression level, as well as its enzymatic activity, measured as verapamil N dealkylation, and genetic polymorphisms from two regions within the CYP3A gene cluster. One region is defined by several variants, mostly located within CYP3A7, the other by a single nucleotide polymorphism in intron 7 of CYP3A4. The effects of these single nucleotide polymorphisms are sex-dependent. For example, female carriers of T alleles of the single nucleotide polymorphism rs4646437C>T in CYP3A4 intron 7 have, respectively, 5.1-fold and 2.7-fold higher expression and activity compared with male T-carriers, but only 2.2-fold and 1.4-fold higher expression and activity compared with males of genotype CC. A regression analysis indicates that the impact of these single nucleotide polymorphisms in men goes beyond the previously reported sex effect. The rs4646437C undergoes positive selection in Caucasians, as evidenced by its relative extended haplotype homozygosity value located within the uppermost percentile of a genome-wide test set of haplotypes in the same 5% frequency bin. CONCLUSIONS: Our findings reconcile the apparent contradiction between the evidence for the influence of the individual genetic makeup on CYP3A4 expression and activity suggested by clinical studies, and the failure to identify the responsible gene variants. PMID- 17465709 TI - Genetic association meets RNA interference: large-scale genomic screens for causation and mechanism of complex diseases. AB - While the genomic era offers great promise for biomedicine in general and for biomarker discovery in particular, it has yet to significantly impact drug target discovery. Meanwhile, despite improvements over the past 20 years in reducing attrition in clinical trials due to adverse drug responses, the pharmaceutical industry continues to be beset by the high rate of attrition of compounds in late stage development, primarily due to the lack of drug efficacy. Clearly, even highly potent drugs with ideal safety and pharmacokinetic profiles will fail to survive clinical trials if the drug target itself is not a key point of intervention for most patients. Genetic association studies and RNA interference are two scaleable genomic approaches that together can address the quality as well as quantity of candidate drug targets. Human genetic information has long been used to identify 'molecular bottlenecks' that can highlight the importance of a gene or pathway at the clinical level. The recent availability of the human HapMap and of high-throughput genotyping platforms now enables more systematic genetic screens for novel, clinically-relevant drug targets. In addition, RNA interference can help dissect the molecular role of a candidate drug target in preclinical model systems in vitro and in vivo. Wider applicability of RNA interference methods will closely follow continued progress on efficient delivery into appropriate cell models and target tissues. PMID- 17465710 TI - Adducin polymorphisms and the treatment of hypertension. AB - Hypertension is an important public health problem affecting more than 50 million individuals in the USA alone. The most common form, essential hypertension, results from the complex interplay between genetic predisposition and environmental influences. Epidemiological, migration, intervention and genetic studies in humans and animals provide very strong evidence of a causal link between high salt intake and high blood pressure. One of the candidate genes for salt-sensitive hypertension is adducin. Adducin is a heterodimeric cytoskeleton protein, the three subunits of which are encoded by genes (ADD1, ADD2 and ADD3) that map to three different chromosomes. A long series of parallel studies in the Milan hypertensive rat strain model of hypertension and humans indicated that an altered adducin function might cause hypertension through enhanced constitutive tubular sodium reabsorption. An example of a prospective efficacy of pharmacogenetics and pharmacogenomics is the detection and impact of adducin polymorphisms on hypertension. In particular, the selective advantages of diuretics in preventing myocardial infarction and stroke over other antihypertensive therapies that produce a similar blood pressure reduction in carriers of the mutated adducin may support new strategies aimed at optimizing the use of new antihypertensive agents for the prevention of hypertension associated organ damage. PMID- 17465711 TI - Key aspects of analyzing microarray gene-expression data. AB - One major challenge with the use of microarray technology is the analysis of massive amounts of gene-expression data for various applications. This review addresses the key aspects of the microarray gene-expression data analysis for the two most common objectives: class comparison and class prediction. Class comparison mainly aims to select which genes are differentially expressed across experimental conditions. Gene selection is separated into two steps: gene ranking and assigning a significance level. Class prediction uses expression profiling analysis to develop a prediction model for patient selection, diagnostic prediction or prognostic classification. Development of a prediction model involves two components: model building and performance assessment. It also describes two additional data analysis methods: gene-class testing and multiple ordering criteria. PMID- 17465712 TI - Gene therapy to fight infection in skin transplants. PMID- 17465714 TI - Pharmacogenetics and stomach cancer: an update. AB - Although new drugs and association regimens have been used in recent years, the chemotherapeutic outcome for gastric cancer is still poor and improvement in patient survival is not satisfactory. Pharmacogenetics could represent a useful approach to optimize chemotherapeutic treatments in order to identify individuals that are true candidates for clinical benefits from therapy, avoiding the development of severe side effects. The most recent update regarding gastric cancer pharmacogenetics highlights a prominent role of genetic polymorphisms of thymidylate synthase and glutathione S-transferase in the pharmacological treatment with commonly used drugs, such as 5-fluorouracil and platinum derivatives. In order to validate the genetic markers, further larger scale and controlled studies are required. A future challenge is represented by the introduction of targeted therapy in gastric cancer treatment, with the potential emerging tool of pharmacogenetic impact on this field. PMID- 17465713 TI - Identifying the genotype behind the phenotype: a role model found in VKORC1 and its association with warfarin dosing. AB - Genotype-phenotype studies in pharmacogenomics promise to identify the genetic factors that contribute substantially to variation in individual drug response. While most genetic association studies have failed to deliver this promise, several recent examples serve as a reminder that these associations do exist and can be identified when investigated using well-designed studies. Here, we describe the path taken to identify the association between common vitamin K epoxide reductase complex subunit 1 genetic variation and warfarin dosing in patients. We also describe the key elements that led the way, such as definition of the phenotype, confirmation of a genetic component, determination of biological plausibility and selection of genetic polymorphisms. We also describe several avenues that are yet to be explored for the specific vitamin K epoxide reductase complex subunit 1 warfarin example that can also be generalized as future directions for many genetic association studies in pharmacogenomics. These future avenues will be best explored using diverse approaches encompassing clinical, statistical and genomic methods currently being developed for genotype phenotype studies in human populations. PMID- 17465715 TI - How can the emerging patient-centric health records lower costs in pharmacogenomics? AB - The costs of clinical trials in which the subjects' inherited and somatic genetic variations are taken into consideration are higher than those trials where no genetic data are used. The higher costs are due to increased complexity in exploring clinical and genomic factors, as well as the need to coordinate the various outputs of disparate entities, such as clinical and genetic laboratories, and infer significant associations between genotype and phenotype observations. Environment, diet and lifestyle factors, in addition to clinical data, can all affect a person's response to medicines and thus there is a need to obtain the complete health history of a subject enrolled in a clinical trial in order to assert the right genotype-phenotype associations. Such complete histories can be made available through the emerging patient-centric health records, and challenges concerning these records, such as sustainability, usability and governance, are discussed in this paper. PMID- 17465717 TI - Genetic nondiscrimination legislation: a critical prerequisite for pharmacogenomics data sharing. PMID- 17465718 TI - New solid tumor targets for therapeutic monoclonal antibodies. AB - Monoclonal antibodies are a rapidly growing class of drugs used for therapy of human cancers and other diseases. They can be used effectively to target tumor specific molecules and thereby modulate key signaling pathways that play a role in tumor growth, survival and metastasis. Clinical success of novel antibodies has stimulated great interest in the promise of antibody therapeutics for cancer. In this editorial, the author describes three key characteristics that define an ideal target antigen for a therapeutic antibody. PMID- 17465719 TI - Targeting the PI3K and MAPK pathways to treat Kaposi's-sarcoma-associated herpes virus infection and pathogenesis. AB - Cells require the ability to appropriately respond to signals in their extracellular environment. To initiate, inhibit and control these processes, the cell has developed a complex network of signaling cascades. The phosphatidylinositol 3-kinase (PI3K) and mitogen-activated protein kinase (MAPK) signaling pathways regulate several responses including mitosis, apoptosis, motility, proliferation, differentiation and many others. It is not surprising, therefore, that many viruses target the PI3K and MAPK pathways as a means to manipulate cellular function. Recently, Kaposi's sarcoma-associated herpes virus (KSHV) has been added to the list. KSHV manipulates the PI3K and MAPK pathways to control such divergent processes as cell survival, cellular migration, immune responses, and to control its own reactivation and lytic replication. Manipulation of the PI3K and MAPK pathways also plays a role in malignant transformation. Here, the authors review the potential to target the PI3K and MAPK signaling pathways to inhibit KSHV infection and pathogenesis. PMID- 17465720 TI - IL-19 and IL-20: two novel cytokines with importance in inflammatory diseases. AB - IL-19 and IL-20 are two cytokines that were discovered in 2000 and 2001, respectively. Based on the structure and location of their genes, their primary and secondary protein structures and the used receptor complexes, they were classified with IL-10, IL-22, IL-24, IL-26, IL-28 and IL-29 in the IL-10 family of cytokines, and form a subgroup with IL-24 within this family. IL-19 and IL-20 are produced by monocytes as well as non-immune tissue cells under inflammatory conditions. IL-19 and IL-20 act via a receptor complex that consists of the IL 20R1 and IL-20R2 chains. IL-20 is additionally able to signal via a second receptor complex (IL-22R1/IL-20R2). It is controversial whether or not IL-19 and IL-20 regulate the function of immune cells. However, the expression of their receptors aliments the perception that the cells of the skin, lungs and reproductive organs as well as various glands are major targets of these mediators. Results from animal experiments and massively increased expression of these mediators in human inflamed tissues support the assumption that they play an important role in the pathogenesis of a few inflammatory diseases. For this reason, the authors have reviewed the facts known at present regarding these cytokines and postulate that IL-19 and IL-20 are pharmacologically interesting distal elements of an inflammatory cascade. PMID- 17465721 TI - The IL-6/sIL-6R complex as a novel target for therapeutic approaches. AB - IL-6 plays a pivotal role in immune responses and certain oncologic conditions. The intense investigation of its biological activity and function led to the discovery of two different IL-6-driven signalling pathways. Binding to the membrane-bound IL-6 receptor (mIL-6R, CD126) causes the recruitment of two gp130 co-receptor molecules (CD130) and the activation of intracellular signalling cascades via gp130. Although this classical pathway is mainly limited to hepatocytes, neutrophils, monocytes/macrophages and certain other leukocyte populations, which express IL-6R on their surface, an alternative mechanism has also been described. Proteolytic cleavage of the mIL-6R protein or translation from alternatively spliced mRNA leads to the generation of a soluble form of the IL-6R (sIL-6R), which is likewise able to bind to IL-6. The resulting IL-6/sIL-6R complex is also capable of binding to gp130 and inducing intracellular signalling. Through this so-called 'trans-signalling' mechanism, IL-6 is able to stimulate cells that lack an endogenous mIL-6R. High levels of IL-6 and sIL-6R have been reported in several chronic inflammatory and autoimmune diseases as well as in cancer. Preclinical animal disease models have provided strong evidence that specific blockade of IL-6-regulated signalling pathways represents a promising approach for the therapy of these diseases. An optimised variant of the recently described fusion protein sgp30Fc is now heading towards its clinical evaluation. PMID- 17465722 TI - New therapeutic targets in atrophic age-related macular degeneration. AB - Age-related macular degeneration (AMD) is the leading cause of blindness in developed countries. There is no effective treatment for the most prevalent atrophic (dry) form of AMD. Atrophic AMD is triggered by abnormalities in the retinal pigment epithelium (RPE) that lies beneath the photoreceptor cells and normally provides critical metabolic support to these light-sensing cells. Secondary to RPE dysfunction, macular rods and cones degenerate leading to the irreversible loss of vision. Oxidative stress, formation of drusen, accumulation of lipofuscin, local inflammation and reactive gliosis represent the pathologic processes implicated in pathogenesis of atrophic AMD. This review discusses potential target areas for small-molecule and biologic intervention, which may lead to development of new therapeutic treatments for atrophic AMD. PMID- 17465723 TI - Multifaceted targeting in cancer: the recent cell death players meet the usual oncogene suspects. AB - Recent complicated advances towards the blueprinting of the altered molecular networks that lie behind cancer development have paved the way for targeted therapy in cancer. This directed a significant part of the research community to the development of specialized targeted agents, many of which are already available or in clinical trials. The prospect of patient-tailored therapeutic strategies, although very close to becoming a reality also raises the level of complexity of the therapeutic approach. This review summarizes the functions, in vivo expression patterns and aberrations of factors presently targeted or representing potential targets by therapeutic agents, focusing on those implicated in death receptor-induced apoptosis. The authors overview the regulation of these factors and death receptor-induced apoptosis by classical oncogenes (e.g., RAS, MYC, HER2) and their effectors/regulators, most of which are also being targeted. In addition, the importance of orthologic systemic approaches in future patient-tailored therapies are discussed. PMID- 17465724 TI - Fatty acid receptors as new therapeutic targets for diabetes. AB - G-protein-coupled receptors (GPCRs) are key regulators of several physiological functions. Their roles in cellular signal transduction have made them the target for majority of all currently prescribed drugs. Additionally, there are many orphan GPCRs that provide potential novel therapeutic targets. Several GPCRs are involved in metabolic regulation and glucose homeostasis such as GLP-1 receptor, glucagon receptor, adiponectin receptor and so on. Recently, free fatty acids (FFAs) have been demonstrated as ligands for orphan GPCRs and have been proposed to play a critical role in physiological glucose homeostasis. GPR40 and GPR120 are activated by medium and long-chain FFAs, whereas GPR41 and GPR43 can be activated by short-chain FFAs. GPR40, which is preferentially expressed in pancreatic beta-cells, mediates the majority of the effects of FFAs on insulin secretion. In this review, these findings and also critical analysis of these GPCRs as novel targets for diabetes are discussed. PMID- 17465725 TI - Targeting signal transduction in pancreatic cancer treatment. AB - Pancreatic cancer is a lethal disease with a 5-year survival rate of 4%. The only opportunity for improved survival continues to be complete surgical resection for those with localized disease. Although chemotherapeutic options are limited for the few patients with resectable disease, this problem is even more magnified in the majority (85%) of patients with unresectable or metastastic disease. Therefore, there is an urgent need for improved therapeutic options. The recent success of inhibitors of signal transduction for the treatment of other cancers supports the need to identify and validate aberrant signaling pathways important for pancreatic tumor growth. This review focuses on the validation of specific signaling networks and the present status of inhibitors of these pathways as therapeutic approaches for pancreatic cancer treatment. PMID- 17465726 TI - Nicotinamide adenine dinucleotide metabolism as an attractive target for drug discovery. AB - Nicotinamide adenine dinucleotide (NAD(+)) has crucial roles in many cellular processes, both as a coenzyme for redox reactions and as a substrate to donate ADP-ribose units. Enzymes involved in NAD(+) metabolism are attractive targets for drug discovery against a variety of human diseases, including cancer, multiple sclerosis, neurodegeneration and Huntington's disease. A small-molecule inhibitor of nicotinamide phosphoribosyltransferase, an enzyme in the salvage pathway of NAD(+) biosynthesis, is presently in clinical trials against cancer. An analog of a kynurenine pathway intermediate is efficacious against multiple sclerosis in an animal model. Indoleamine 2,3-dioxygenase plays an important role in immune evasion by cancer cells and other disease processes. Inhibitors against kynurenine 3-hydroxylase can reduce the production of neurotoxic metabolites while increasing the production of neuroprotective compounds. This review summarizes the existing knowledge on NAD(+) metabolic enzymes, with emphasis on their relevance for drug discovery. PMID- 17465727 TI - The potential for phospholipase D as a new therapeutic target. AB - Mammalian phospholipase D (PLD), a signal transduction-activated enzyme, hydrolyzes phosphatidylcholine to generate the lipid second messenger phosphatidic acid (PA) and choline. Genetic and pharmacological methods have implicated PLD and its product PA in a wide variety of cellular processes including vesicle trafficking, receptor signaling, cell proliferation and survival. Dysregulation of these cell biologic processes occurs in a diverse range of illnesses including cancer. This review summarizes PLD regulation and function and highlights its potential as a therapeutic target in disease settings. PMID- 17465728 TI - Towards the discovery of novel T-type calcium channel blockers. AB - Despite their presence in many tissues and their potential implication in various disease states, low-voltage activated T-type calcium channels (T-channels) have only recently become targets of interest. Unfortunately, the lack of selective T channel blockers has hampered further characterisation of these channels. The recent availability of cloned T-channels, the Ca(V)3 proteins, facilitates identification of novel T-channel blockers. Also, studies performed in knockout animals have fostered novel interest. Selective inhibition of T-channels may have clinical importance in cardiovascular diseases, some forms of epilepsy, sleep disorders, pain and possibly cancer. This review focuses on novel research approaches to discover potent and selective T-channel modulators. These molecules may be potential drugs for treating human diseases, as well as important tools to decipher the physiological role of these channels. PMID- 17465729 TI - Regenerative medicine bioprocessing: the need to learn from the experience of other fields. PMID- 17465732 TI - Neural induction and neural stem cell development. AB - Embryonic stem (ES) cells are a pluripotent and renewable cellular resource with tremendous potential for broad applications in regenerative medicine. Arguably the most important consideration for stem cell-based therapies is the ability to precisely direct the differentiation of stem cells along a preferred cellular lineage. During development, lineage commitment is a multistep process requiring the activation and repression of sets of genes at various stages, from an ES cell identity to a tissue-specific stem cell identity and beyond. Thus, the challenge is to ensure that the pattern of genomic regulation is recapitulated during the in vitro differentiation of ES cells into stem/progenitor cells of the appropriate tissue in a robust, predictable and stable manner. To address this issue, we must understand the ontogeny of tissue-specific stem cells during normal embryogenesis and compare the ontogeny of tissue-specific stem cells in ES cell models. Here, we discuss the issue of directed differentiation of pluripotent ES cells into neural stem cells, which is fundamentally linked to two early events in the development of the mammalian nervous system: the 'decision' of the ectoderm to acquire a neural identity (neural determination) and the origin of neural stem cells within this neural-committed population of cells. A clearer understanding of the molecular and cellular mechanisms that govern mammalian neural cell fate determination will lead to improved ES technology applications in neural regeneration. PMID- 17465733 TI - History of regenerative medicine: looking backwards to move forwards. AB - If the song by the international popstar Sting is to be relied upon, 'History will teach us nothing'. At the risk of contradicting a one-time schoolteacher, I believe the reverse is true for regenerative medicine. In fact, I think we dismiss the past at our peril. In this review, I aim to trace the history of regenerative medicine to date. I will examine parallels with other areas of medicine and show how commercial, technical and socio/economic factors have influenced the pace and direction of the sector's evolution. I will discuss how, by learning from the past, those involved in regenerative medicine are reinventing their sector for the better. In conclusion, I will evaluate the current state of the industry, suggest what the future may hold and explain why I believe regenerative medicine is about to 'come of age'. PMID- 17465734 TI - China and the global stem cell bioeconomy: an emerging political strategy? AB - There is a growing consensus, amongst policy analysts and scientists alike, that China is likely to play a key role in the scientific, clinical and commercial development of stem cell research. However, to date, there exist few detailed analyses of China's current investment in the field. After introducing the UK's recent political strategy on stem cell science, this article develops an in-depth discussion of the formal organization of China's research and development in the area, as well as its rapidly evolving commercial, regulatory and ethical environment. From here, we go on to assess the probability of China's emergence as a global player in the increasingly internationalized business of stem cell biomedicine. PMID- 17465735 TI - Taking tissue-engineering principles into theater: augmentation of impacted allograft with human bone marrow stromal cells. AB - Human bone marrow contains bone progenitor cells that arise from multipotent mesenchymal stem cells. Seeding bone progenitor cells onto a scaffold can produce a 3D living composite with significant mechanical and biological potential. This article details laboratory and clinical findings from two clinical cases, where different proximal femoral conditions were treated using impacted allograft augmented with marrow-derived autogenous progenitor cells. Autologous bone marrow was seeded onto highly washed morselized allograft and impacted. Samples of the impacted graft were also taken for ex vivo analysis. Both patients made an uncomplicated clinical recovery. Imaging confirmed defect filling with encouraging initial graft incorporation. Histochemical and alkaline phosphatase staining demonstrated that a live composite graft with osteogenic activity had been introduced into the defects. These studies demonstrate that marrow-derived cells can adhere to highly washed morselized allograft, survive the impaction process and proliferate with an osteoblastic phenotype, thus creating a living composite. PMID- 17465737 TI - Patenting of stem cells. AB - Investors in any new technology are concerned to protect their investment, a key part of such protection being the availability of patent protection. Stem cells, human embryonic stem cells in particular, are a highly controversial area, and this controversy extends to the patenting of stem cells. In this article, the legal issues affecting patenting of stem cell technology in the USA and Europe are reviewed. The types of patents that have been granted are also considered, as an illustration of the protection that can be obtained. Finally, the overall trends in patent filings are discussed, to identify key aspects of the patent landscape. PMID- 17465738 TI - New guidance on codes of practice, standardized methods and regulations for cell based therapeutics. AB - Consultations with members of the UK regenerative medicine community have identified the need for greater overall clarity regarding the current legislation, guidance and standards to be used. To facilitate this, the UK Department of Trade and Industry commissioned the British Standards Institution to develop guidance on the product pathway from cell/tissue donation to clinical trial for cell-based therapies within the UK and outline the accompanying legislation, codes of practice, guidance documents and standards for this process. This document will be aimed at a cross-section of end users from academic groups through to small and medium enterprises, larger industrial companies and the general public. PMID- 17465739 TI - Regulation of tissue-engineered products in the European Union: where are we heading? AB - The emergence of tissue-engineered products (TEPs) raises a standard question for regulators: is the existing regulatory regime appropriate or is there a case for a new regulatory framework? In the USA, the FDA has developed a risk-based approach to TEPs, whereas in Europe, a common regulatory strategy for these products has not yet been implemented. In order to fill this perceived gap, member states have set up domestic rules, which has led to an unclear and patchy regulatory situation. The Regulation on Advanced Therapy Medicinal Products, voted on by the European Commission in November 2005, has been developed by European Union regulators to provide the necessary framework to regulate TEPs. As the text is still to be discussed and to be passed, many concerns have been raised regarding the appropriateness of the proposed framework. PMID- 17465740 TI - Stem cell therapy delivery: treading the regulatory tightrope. AB - The concept of stem cell therapy has engaged the attention of the public and scientists alike. Intensive research effort is focused upon understanding the biology and therapeutic potential of embryonic and adult stem cells, with the eventual goal of treating such pathologies as Parkinson's disease, diabetes, neurological injury and degenerations and cancer. Ex vivo expansion and transplantation of limbal epithelial stem cells to the corneas to treat blinding ocular surface disease was one of the first stem cell therapies to successfully reach the clinic. However, limbal epithelial stem cell research and therapy delivery has remained largely within the noncommercial academic clinician scientist environment from which it was originally pioneered. In our experience, gaining regulatory approval has been as great a hurdle as surmounting the scientific challenges of stem cell therapy. Based upon our model of delivering 'accredited' limbal epithelial stem cell therapy to patients in compliance with Good Manufacturing Practice and the new European Union Tissues and Cells Directive, we address the key regulatory questions. This may help colleagues who are developing innovative academic research-driven stem cell therapies regarding donor consent, raw materials, quality assurance, laboratory specification, indemnity and funding. PMID- 17465741 TI - Stem Cell Sciences plc. AB - Stem Cell Sciences' core objective is to develop safe and effective stem cell based therapies for currently incurable diseases. In order to achieve this goal, Stem Cell Sciences recognizes the need for multiple technologies and a globally integrated stem cell initiative. The key challenges for the successful application of stem cells in the clinic is the need for a reproducible supply of pure, fully characterized stem cells that have been grown in suitable conditions for use in the clinic. PMID- 17465742 TI - Interspecies SCNT-derived human embryos--a new way forward for regenerative medicine. PMID- 17465743 TI - Mesenchymal stem cells for the treatment of heart disease. PMID- 17465746 TI - Engineering the heart piece by piece: state of the art in cardiac tissue engineering. AB - According to the National Transplant Society, more than 7000 Americans in need of organs die every year owing to a lack of lifesaving organs. Bioengineering 3D organs in vitro for subsequent implantation may provide a solution to this problem. The field of tissue engineering in its most rudimentary form is focused on the developed of transplantable organ substitutes in the laboratory. The objective of this article is to introduce important technological hurdles in the field of cardiac tissue engineering. This review starts with an overview of tissue engineering, followed by an introduction to the field of cardiovascular tissue engineering and finally summarizes some of the key advances in cardiac tissue engineering; specific topics discussed in this article include cell sourcing and biomaterials, in vitro models of cardiac muscle and bioreactors. The article concludes with thoughts on the utility of tissue-engineering models in basic research as well as critical technological hurdles that need to be addressed in the future. PMID- 17465747 TI - Meeting the needs of monitoring in tissue engineering. AB - Tissue engineering is a rapidly growing field that aims to develop biological substitutes that restore, maintain or improve tissue function. The focus of research to date has been the underlying biology required for tissue-engineered therapies. However, as tissue-engineered products reach the marketplace, there is a pressing need for an improved understanding of the engineering and economic issues associated with them. This is motivated by the lack of commercial viability of many of the initial therapies that have been produced. It has been suggested in the literature that this is partly due to poor process and system design in tissue production, as well as a lack of process monitoring and control. This review argues that principles of design, measurement and process monitoring from the physical sciences are needed to move tissue engineering forward, and that much of the technology needed to realize this is already available. PMID- 17465748 TI - Hepatocyte growth factor: a regenerative drug for acute hepatitis and liver cirrhosis. AB - Liver cirrhosis is a major cause of morbidity worldwide and is characterized by the loss of hepatocytes with interstitial fibrosis. In this review, we discuss the potential uses of hepatocyte growth factor for treating hepatic diseases, focusing on the molecular mechanisms whereby hepatocyte growth factor reverses liver cirrhosis. Hepatic myofibroblasts play a central role in the development of liver cirrhosis, while myofibroblasts acquire c-Met. Using a rat model of liver cirrhosis, we recently delineated the direct effect of hepatocyte growth factor toward myofibroblasts: the induction of apoptotic cell death associated with matrix degradation, the inhibition of overproliferation and the suppression of transforming growth factor-beta1 production in myofibroblasts. Hepatocyte growth factor elicits mitogenic, anti-apoptotic and anti-inflammatory functions in hepatocytes, therefore contributing to reversing liver dysfunction. Considering the insufficient production of hepatocyte growth factor is responsible for the manifestation of chronic hepatitis, supplementation with or reinduction of hepatocyte growth factor represents a new strategy for attenuating intractable liver diseases. PMID- 17465749 TI - Stem cell-based therapy for the treatment of Type 1 diabetes mellitus. AB - Diabetes mellitus is the most common metabolic disorder, which occurs in two forms: Type 1 diabetes (juvenile or insulin-dependent diabetes mellitus) and Type 2 diabetes (adult or noninsulin-dependent diabetes mellitus). Type 1 diabetes mellitus is a T-cell-mediated, organ-specific autoimmune disorder, in which the body's own immune system attacks beta-cells and damages them sufficiently resulting in reduced insulin production. To overcome autoimmunity, immunosuppressive therapy, gene therapy, islet cell regeneration or encapsulation of islet cells offer dramatic treatment solutions. At present, efforts for finding ways to replace damaged insulin-secreting beta-cells by implanting new cells is an active field of research. Various therapeutic strategies are under investigation and stem cell-based therapy with the combination of other treatments offers exciting possibilities for the development of treatment for such diseases. In the current review, we focus on stem cells and their potential clinical applications and summarize the recent progress in this field. PMID- 17465750 TI - A panel of tests to standardize the characterization of human embryonic stem cells. AB - Human embryonic stem cells offer a renewable source of a wide range of cell types for use in research and cell-based therapies. Characterizing these cells provides important information about their current state and affords relevant details for subsequent downstream manipulation. Prior to considering therapeutic applications, it is crucial that the cells are surveyed at a genetic and proteomic level during the extensive propagation, expansion and differentiation. Hence, a set of characterization tests to measure stem cell stability and identity--genomic, epigenomic and mitochondrial markers, as well as functional measures of utility, need to be developed. Thus, we outline a plan of standard assays that can be afforded by multiple laboratories to unambiguously test the quality of human embryonic stem cells. In this manuscript, we describe a comprehensive characterization of ReliCell hES1, the only human embryonic stem cell line reported from the Indian subcontinent. Our study employs gene expression analysis using quantitative reverse transcription-polymerase chain reaction and microarray, mitochondrial DNA sequencing, microRNA analysis, immunophenotyping and teratoma formation, in addition to demonstrating its capacity to propagate under feeder-free conditions. PMID- 17465751 TI - Creating prodynorphin-expressing stem cells alerted for a high-throughput of cardiogenic commitment. AB - BACKGROUND: The development of cell therapy for the rescue of damaged heart muscle is a major area of inquiry. Within this context, the establishment of a cardiogenic cell line may remarkably facilitate the molecular dissection of cardiac fate specification, a low-efficiency and still poorly understood process, paving the way for novel approaches in the use of stem cells for cardiac repair. METHODS & RESULTS: We used GTR1 cells, a derivative of mouse R1 embryonic stem cells bearing the puromycin-resistance gene driven by the cardiomyocyte-specific alpha-myosin heavy chain promoter, affording a gene trapping selection of a virtually pure population of embryonic stem cell-derived cardiomyocytes. Third generation lentiviral vectors were used to overexpress the prodynorphin gene, previously shown to orchestrate a dynorphinergic system acting as a major conductor of embryonic stem cell cardiogenesis. Lentiviral prodynorphin transduction remarkably enhanced the transcription of GATA-4 and Nkx-2.5, two cardiac lineage-promoting genes, resulting in a dramatic increase in the number of spontaneously beating cardiomyocytes. Transduced cells also exhibited a subcellular redistribution patterning of protein kinase C-beta, -delta and epsilon, a major requirement in cardiac lineage commitment. This activation resulted from a sustained increase in the transcription of targeted protein kinase C genes. Prodynorphin transduction was selective in nature and failed to activate genes responsible for skeletal myogenesis or neuronal specification. CONCLUSIONS: The cell line developed in this study provides a powerful in vitro model of cardiomyogenesis that may help clarify the cascade of transcriptional activation and signaling networks that push multipotent cells to take on the identity of a cardiac myocyte. PMID- 17465752 TI - Why we should, in fact, pay for egg donation. AB - In this perspective, I shall argue that women who donate eggs solely for human embryonic stem cell research ought to be compensated. My argument rests on three inter-related principles. First, it is important to recruit the healthiest possible egg donors to minimize the risks of donation. This would relieve pressure to donate on those suffering from diseases that might be treatable with stem cell-based therapies, who are likely to be at greater risk from donation. Second, I believe that it is crucial to be pro-active in building representative stem cell banks, especially in stem cell initiatives paid for, in part, by the public/government. The right of all groups to participate in and benefit from equitable and safe research must be developed for egg donors as for other kinds of research participants. Particular attention should be paid to the opinions and desires of women from historically underserved populations as to how to conduct donations and guide research so as to serve all members of society. Third, reasonable payment would undermine tendencies for domestic and international black and grey egg markets for stem cell research to develop. I then suggest replacing the question of compensation with the question of harm mitigation as the central donor protection issue. PMID- 17465754 TI - Advanced Cell Technology, Inc. AB - Advanced Cell Technology, Inc. (OTCBB: ACTC) is a biotechnology company applying novel human embryonic stem cell technologies in the emerging field of regenerative medicine. We believe that regenerative medicine has the potential to revolutionize the field by enabling scientists to produce human cells of any kind for use in a wide array of therapies. PMID- 17465755 TI - Proposal for a universal minimum information convention for the reporting on the derivation of human embryonic stem cell lines. PMID- 17465758 TI - Physical activity-mediated functional recovery after spinal cord injury: potential roles of neural stem cells. AB - As data elucidating the complexity of spinal cord injury pathophysiology emerge, it is increasingly being recognized that successful repair will probably require a multifaceted approach that combines tactics from various biomedical disciplines, including pharmacology, cell transplantation, gene therapy and material sciences. Recently, new evidence highlighting the benefit of physical activity and rehabilitation interventions during the post-injury phase has provided novel possibilities in realizing effective repair after spinal cord injury. However, before a comprehensive therapeutic strategy that optimally utilizes the benefits of each of these disciplines can be designed, the basic mechanisms by which these various interventions act must be thoroughly explored and important synergistic and antagonistic interactions identified. In examining the mechanisms by which physical activity-based functional recovery after spinal cord injury is effected, endogenous neural stem cells, in our opinion, engender a potentially key role. Multipotent neural stem cells possess many faculties that abet recovery, including the ability to assess the local microenvironment and deliver biofactors that promote neuroplasticity and regeneration, as well as the potential to replenish damaged or eradicated cellular elements. Encouragingly, the functional recovery owing to physical activity-based therapies appears relatively robust, even when therapy is initiated in the chronic stage of spinal cord injury. In this article, we review experimental outcomes related to our hypothesis that endogenous neural stem cells mediate the functional recovery noted in spinal cord injury following physical activity-based treatments. Overall, the data advocates the incorporation of increased physical activity as a component of the multidimensional treatment of spinal cord injury and underscores the critical need to employ research-based mechanistic approaches for developing future advances in the rehabilitation of neurological injury and disorders. PMID- 17465759 TI - Umbilical cord-derived stem cells for tissue therapy: current and future uses. AB - Organ and tissue transplants provide a means to correct disease but are limited, mostly owing to the lack of available donor tissue. Tissue matching and speed of procurement are important parameters that must be met for a successful graft, however the lack of available donors leads to expanding waiting lists and suboptimal human leukocyte antigen-matching, often leading to reduced transplant success. The discovery of embryonic stem cells and tissue-specific stem cells has provided hope for many patients. Stem cell-based therapy has provided possible new sources of human leukocyte antigen-matched tissue but, before routine clinical application of stem cells becomes a reality, many obstacles must first be overcome. Focusing on umbilical cord blood cells, we discuss some of the challenges that stem cell therapy faces, including obtaining clinically relevant numbers of stem cells and the ability of stem cells to provide for permanent engraftment of multiple tissue types. We discuss possible solutions to these problems, such as in vitro stem cell expansion and the differentiation potential of tissue-specific stem cells. PMID- 17465760 TI - Biocompatibility and the efficacy of medical implants. AB - Biocompatibility is key to the performance of any material in a biological environment. This review outlines current opinion on the factors that lead to biocompatibility and focuses on the interactions that occur at the interface between material and environment. The sequence of events, from protein adsorption, cell attachment and behavior, to biocompatibility, is traced. Although these processes are studied and reported widely, there is, as yet, little published evidence that implant biocompatibility can be enhanced in the long term by surface engineering. This lack of evidence does not necessarily imply a lack of effect, but may be ascribed to a lack of robust characterization and poor modeling of the implant environment. PMID- 17465761 TI - Commercial development of stem cell technology: lessons from the past, strategies for the future. AB - This paper presents historical and contemporary survey data on the commercial development of stem cell technology from the 1990s to the present day. We describe the first wave of industrial investment in hematopoietic stem cells during the 1990s and contrast this with the more recent expansion of the sector. In particular, we explore the cell types used, diseases targeted and business models adopted by firms. We conclude, by arguing that the commercial prospects for stem cell technologies remain highly uncertain and that innovative public policies should be adopted to prevent 'market failure'. PMID- 17465762 TI - Clinical experience using cultured epithelial autografts leads to an alternative methodology for transferring skin cells from the laboratory to the patient. AB - We report a 10-year audit using cultured epithelial autografts (CEAs) for patients with extensive burns. Clinical take using CEAs averaged only 45% (as has been reported by others) but over half of all cells cultured for these patients had to be discarded owing to difficulties of timing the production of CEA sheets to the needs of the patients. CEAs could not be used until they had reached confluence and formed an integrated sheet, which took, on average, 12 days. However, once formed, they needed to be used within 2-3 days or they lost the ability to attach to wound beds. In response to this we developed a simpler carrier dressing methodology for transferring cultured subconfluent keratinocytes from the laboratory to the wound bed. This methodology offers an increase in speed of delivery but its major contribution is the greater flexibility in timing the transfer of cells from the laboratory to the changing needs of the patients. PMID- 17465763 TI - Emotion in political discourse: contrasting approaches to stem cell governance in the USA, UK, Israel and Germany. AB - In August 2004, Stojkovic and Murdoch from the University of Newcastle upon Tyne, UK, were granted the UK's first license to create human embryonic stem cells (hESCs) using cell nuclear replacement. While this news made headlines around the globe, a spokesman for the German Ministry of Research warned scientists in his country of the illegality of advising their English colleagues on hESC research. Meanwhile, US Members of Congress had asked President Bush to revoke his decision to limit federal funding to research on a limited number of hESC lines created before 9 August, 2001 (a decision that he confirmed in July 2006, while nonfederally funded research on hESC continues to be unrestricted). In Israel, where hESC research is legal and has never been a contested political issue, a bioethicist argued that, in light of the potential to alleviate human suffering, "banning research is against human dignity". How can such striking differences in the regulation of hESC research be explained? PMID- 17465764 TI - Stem cell research and the ethics of transparency. AB - Stem cell research represents a field of scientific inquiry subject to intense public and political attention and debate worldwide. There are several reasons for this. First, it is a research endeavor surrounded by great expectations of future therapeutic benefits. Some of these expectations seem to be well founded, while others originate from adult and embryonic research enthusiasts having hyped their case. A second reason why this represents a field of public and political attention relates to deeply felt concerns regarding the moral justifiability of sacrificing potential human lives for research. The aim of this paper is to discuss different ways of making the performance of international embryonic stem cell research more transparent, and of unveiling the need for more open-minded dialog concerning the ethical costs of this research endeavor. PMID- 17465766 TI - Theregen, inc. AB - Theregen, Inc. is a regenerative medicine company based in San Francisco, California, USA, that develops cell-based therapies for patients with cardiovascular and vascular disease. The company is conducting a Phase I human clinical safety trial at two US sites for its lead product, Anginera, a cell based epicardial patch therapy. Anginera is designed to promote blood vessel growth in damaged cardiac tissue, stimulate wound healing and, potentially, to restore cardiac function in patients with heart failure. PMID- 17465768 TI - European approval system for advanced therapies: good news for patients and innovators alike. PMID- 17465769 TI - Stem cells for multiple sclerosis: promises and reality. PMID- 17465770 TI - Regenerative medicine 2.0. PMID- 17465772 TI - California dreaming? A new start for regenerative medicine in the Golden State. Interview with Dr. Zach Hall. AB - The California Institute for Regenerative Medicine (CIRM) was established in 2004 with the passage of Proposition 71, the California Stem Cell Research and Cures Initiative. The statewide ballot measure, which provided US$3 billion in funding for stem cell research at California universities and research institutions, was approved by California voters, and called for the establishment of an entity to make grants and provide loans for stem cell research, research facilities and other vital research opportunities. Here, Dr Zach Hall, Interim President of the CIRM, outlines the ethos and aspirations of the CIRM to Regenerative Medicine. Dr Hall trained as a basic neuroscientist and became a faculty member and department chair at the University of California, San Francisco. In 1994, he was appointed Director of National Institute of Neurological Disorders and Stroke within the National Institutes of Health, and was responsible for a research program that awarded more than US$500 million a year in grants and contracts. Since that time, he has held senior positions in research administration within both the University of California, San Francisco, where he was Executive Vice Chancellor, and the University of Southern California. Full information about the CIRM can be found at www.cirm.ca.gov. PMID- 17465774 TI - Bone marrow-derived stem cells in neurological diseases: stones or masons? AB - In spite of the commonly held belief that 'the brain does not regenerate', it is now accepted that postnatal neurogenesis does occur. Thus, one wonders whether cellular-replacement therapy might be used to heal the brain in diseases caused by neuronal cell loss. The existence of neural stem cells has been demonstrated by many scientists and is now generally accepted. The exact role of these cells, how their numbers are regulated and how they participate in CNS and spinal cord regeneration in postnatal life are still not well known. There are many reviews summarizing work on these cells; consequently, I will focus instead on other cells that may participate in postnatal neurogenesis: bone marrow-derived stem cells. The possibility that bone marrow-derived stem cells populate the CNS and differentiate into various neural elements is certainly not universally accepted. PMID- 17465775 TI - Protocols for studying adult neurogenesis: insights and recent developments. AB - The first evidence that neurogenesis occurs in the adult brain was reported in rodents in the early 1960s, using [(3)H]-thymidine autoradiography. In the 1980s and 90s, the advent of new techniques and protocols for studying cell proliferation in situ, and particularly bromodeoxyuridine labeling, helped to confirm that neurogenesis occurs in the adult brain and neural stem cells reside in the adult CNS, including in humans. Bromodeoxyuridine labeling is currently the method most commonly used for studying neurogenesis in the adult brain. However, this procedure is not without limitations, and controversies. In this article, I will review recent protocols for studying adult neurogenesis, particularly new protocols for studying cell kinetics and cell proliferative history, using halopyrimidines. I will review these techniques, and discuss their implications for the field of adult neurogenesis. PMID- 17465776 TI - Role of a paracrine action of mesenchymal stem cells in the improvement of left ventricular function after coronary artery occlusion in rats. AB - PURPOSE: We aimed to determine whether soluble factors released by cultured mesenchymal stem cells (MSCs) improved cardiac function in an experimental model of myocardial infarction. METHODS: MSCs were cultured in fresh medium. The conditioned medium, which contained factors secreted by MSCs, was collected after 4 days of culture. Fischer rats with 1-week-old myocardial infarction were divided into four groups that received: saline (n = 12); fresh medium (n = 10); conditioned medium (n = 8); or 2 million MSCs in fresh medium (n = 10) by direct intramyocardial injection. A total of 4 weeks later, left ventricular (LV) function was assessed by LV angiogram and by LV catheterization. Hearts were processed for histology. RESULTS: Before treatment, LV angiogram assessment demonstrated that the baseline LV function was comparable among the four groups. At 4 weeks after treatment, LV angiogram and LV catheterization showed that LV ejection fraction was better in the fresh medium (49.5 +/- 1.0%), conditioned medium (48.5 +/- 2.1%) and MSCs groups (49.9 +/- 4.2%) than in the saline group (43.7 +/- 1.2%; p < 0.05). There were no significant differences in heart rate, blood pressure, postmortem LV volume, infarct size or septum thickness among the groups. The scar thickness was similar in the saline (395 +/- 31 microm), fresh medium (404 +/- 30 microm) and conditioned medium (397 +/- 34 microm) groups, but significantly thicker in the MSCs group (560 +/- 51 microm; p < 0.05). CONCLUSION: Fresh medium, conditioned medium and MSC injection all improved LV function at 4 weeks after treatment compared with saline treatment in a rat myocardial infarct model; only MSCs increased wall thickness. Since the culture medium contains nutrients and bovine serum, the roles of the soluble factors released by MSCs might be masked. The effect of these nutrients needs further investigation. PMID- 17465777 TI - Endogenous neurogenesis in the human brain following cerebral infarction. AB - Increased endogenous neurogenesis has a significant regenerative role in many experimental models of cerebrovascular diseases, but there have been very few studies in humans. We therefore examined whether there was evidence of altered endogenous neurogenesis in an 84-year-old patient who suffered a cerebrovascular accident 1 week prior to death. Using antibodies that specifically label neural stem/neural progenitor cells, we examined the presence of immunopositive cells around and distant from the infarcted area, and compared this with a control, age matched individual. Interestingly, a large number of neural stem cells, vascular endothelial growth factor-immunopositive cells and new blood vessels were observed only around the region of infarction, and none in the corresponding brain areas of the healthy control. In addition, an increased number of neural stem cells was observed in the neurogenic region of the lateral ventricle wall. Our results suggest increased endogenous neurogenesis associated with neovascularization and migration of newly-formed cells towards a region of cerebrovascular damage in the adult human brain and highlight possible mechanisms underlying this process. PMID- 17465778 TI - Stem cell science in India: emerging economies and the politics of globalization. AB - The globalization of stem cell science is increasingly being shaped by the emerging economies of the Asia/Pacific region. Undaunted and unhampered by the more established views of the commercialization of science, countries such as India are constructing models of innovation, policies and patterns of investment that challenge such orthodoxies. This report examines the position of India within the globalization of stem cell science, its adjustments to the developing knowledge market in this field and its particular contribution to the likely future of this promising bioeconomy. PMID- 17465779 TI - Tissue engineering today, not tomorrow. PMID- 17465780 TI - Genzyme: 15 years of cell and gene therapy research. AB - Finding solutions for patients with unmet needs is at the heart of Genzyme's innovative medical research. While small molecules and enzyme-replacement therapies have made significant strides in improving patients' quality of life and expectancy, the solutions to many conditions, such as heart disease or CNS diseases, rely on the capacity of the body to regenerate cells. Cell and gene therapy lend themselves well to providing solutions for these illnesses with no current cure, by harnessing the body's natural ability to heal. Promising research in these fields continues to evolve and constitutes a long-term investment in addressing serious unmet medical needs in various therapeutic areas. Over the past 15 years, Genzyme has established itself as a pioneer in shaping the research and commercial application of cell and gene therapy. Genzyme's first commercially available innovative cell therapies--Carticel and Epicel--have provided a solid foundation to move into new, cutting-edge areas. PMID- 17465781 TI - Clinical applications of stem cell therapy--the pros and cons of stem cell sources. PMID- 17465782 TI - The time has come to engineer tissues and not just tissue engineer. PMID- 17465785 TI - Human embryonic stem cells: the battle between self-renewal and differentiation. AB - Human embryonic stem cells are pluripotent cells derived from the inner cell mass of blastocyst-stage embryos. These cells possess two unique properties: an indefinite self-renewal capacity and pluripotency, the ability to differentiate to cells from the three germ layers. The pathways governing self-renewal and pluripotency are currently under intensive research. Much effort is devoted to the establishment of feeder-free cultures by elucidation of the cytokines and growth factors required for cell propagation. These seem thus far, to be distinct from those required by mouse embryonic stem cells. In addition, transcriptional regulators unique to embryonic stem cells seem to govern the pluripotent state. These transcriptional regulators determine cell fate, and decide whether the cell will remain pluripotent or differentiate. Together, the understanding of the exogenous and endogenous factors determining cell fate will facilitate the use of these cells in cell-based therapies and will allow understanding of early developmental processes. PMID- 17465786 TI - Conversion of embryonic stem cells into pancreatic beta-cell surrogates guided by ontogeny. AB - Cellular therapies to treat Type 1 diabetes are being devised and the use of human embryonic stem cells (hESCs) offers a solution to the issue of supply, because hESCs can be maintained in a pluripotent state indefinitely. Furthermore, hESCs have advantages in terms of their plasticity and reduced immunogenicity. Several strategies that have so far been investigated indicate that hESCs are capable of differentiating into insulin producing beta-cell surrogates. However the efficiency of the differentiation procedures used is generally quite low and the cell populations derived are often highly heterogenous. A strategy that appears to have long term potential is to design differentiation procedures based on the ontogeny of the beta-cell. The focus of this strategy is to replicate signaling processes that are known to be involved in the maturation of a beta cell. The earliest pancreatic progenitors found in the developing vertebrate fetus are produced via a process known as gastrulation and form part of the definitive endoderm germ layer. hESCs have recently been differentiated into definitive endoderm with high efficiency using a differentiation procedure that mimics the signaling that occurs during gastrulation and the formation of the definitive endoderm. Subsequent events during pancreas development involve a section of the definitive endoderm forming into pancreatic epithelium, which then branches into the pancreatic mesenchyme to form islet clusters of endocrine cells. A proportion of the endocrine precursor cells within islets develop into insulin producing beta-cells. The challenge currently is to design hESC differentiation procedures that mimic the combined events of these stages of beta cell development. PMID- 17465787 TI - Bone marrow-derived stem cell therapy in ischemic heart disease. AB - Since the first experiments of cell transplantation into the heart were performed in the early 1990s, the identification of adult stem cells has triggered attempts to regenerate damaged heart tissue by cellular transplantation. Until recently, a multitude of adult stem or progenitor cells from various tissues have been proposed to meet this end. Bone marrow in particular has emerged as the most promising source for stem and progenitor cells because, besides being the organ of hematopoietic maintenance, it contains a complex assortment of stem and progenitor cells. A large body of provocative experimental evidence for vascular and myocardial regeneration by these cells has generated further enthusiasm for their use. However, many questions remain unanswered in this new field of research regarding the therapeutic potential and the mechanisms responsible for the observed therapeutic effects. In this review, the authors discuss the therapeutic capacity of currently available representative bone marrow-derived stem and progenitor cells for treating ischemic heart diseases. PMID- 17465788 TI - Magnetic resonance imaging and its role in myocardial regenerative therapy. AB - There has been extensive interest recently in cardiac stem cell therapy. Current research has been hampered by differences in cell type, methods of delivery and efficacy evaluation. In this article we review the use of magnetic resonance imaging in this growing area and argue that it is well suited to all areas of myocardial regeneration: from patient identification, through cell delivery and tracking of appropriately labeled cells, to evaluation of therapeutic effect. Potential future advances are discussed including magnetic resonance imaging guided intervention suites and the use of higher field strength magnets for cell tracking. PMID- 17465789 TI - Are countries that ban human embryonic stem cell research hypocritical? AB - In this paper I analyze whether countries that ban human embryonic stem cell research, but are likely to allow their citizens to be treated with stem cell derived products are guilty of hypocrisy. I conclude that the charge of hypocrisy is much more difficult to sustain than many believe. PMID- 17465790 TI - Cognitive regeneration or enhancement: the ethical issues. AB - The neurological and physiological basis of brain function and disease has been a significant focus of investigation throughout the history of medical research. Recent advances in understanding have led to the development of new treatments for diseases of the brain and defects of cognitive and behavioral function: pharmacological, cell-based and even gene therapy may all provide keys to cognitive regeneration. Such therapies, however, might be applied not only towards restoring brain function in the case of disease but to enhance cognitive function for healthy individuals. The concept of cognitive enhancement raises many ethical issues: whether brain-enhancing treatments should be developed and made available and to whom; and what potential consequences might arise? This paper explores some of the ethical arguments associated with cognitive enhancement and concludes that although the technology involved is as yet uncertain and issues of social equity remain to be addressed, the potential benefit of enhancing human brain function is clear. PMID- 17465791 TI - Hypocellularity and absence of compaction as criteria for embryonic death. AB - BACKGROUND: A precise definition of death is important for the appropriate application of medical resources and the harvesting of tissues for transplantation. For developed humans, life is considered to end when the criteria for brain death are met, but corresponding criteria are lacking for human embryos, and thus, we undertook a natural history study of embryonic death. METHODS: De-identified records of the observations of human embryos in culture were analyzed retrospectively. The embryos were generated by in vitro fertilization for the purpose of reproduction. Cell number and morphology were recorded on embryonic days 2, 3, 5, and 6. Viable embryos (n = 248) were compared to nonviable embryos (n = 444) and the latter were analyzed in subgroups defined by cell number and morphology. RESULTS: Many nonviable embryos (n = 142 out of 444) were hypocellular and lacked compaction on embryonic day 5 (ED5). All of these hypocellular embryos did not progress to compacted morula or normal blastocyst when observed further. No criteria could be discerned for the diagnosis of death on ED3. CONCLUSIONS: Arrested development at the multicellular stage on ED5 indicates an irreversible loss of integrated organic function, and hence, the condition of death for the organism. Approximately a fifth of all embryos generated for in vitro fertilization--heretofore misclassified among the 'nonviable'--are in fact dead on ED5 by our criteria. We propose that the ethical framework currently used for obtaining essential organs from deceased persons for transplantation could be applied to the harvesting of live cells from dead human embryos for the creation of stem cells. PMID- 17465792 TI - The International Society for Stem Cell Research (ISSCR): history and perspectives. AB - The International Society for Stem Cell Research (ISSCR) is an independent, nonprofit organization founded in 2002 to foster professional and public communication and education regarding stem cell research. Under the leadership of prominent stem cell scientists from around the world, the ISSCR membership has grown exponentially, creating a diverse, international community of researchers who have expertise in many facets of stem cell biology. The ISSCR is dedicated to promoting the exchange of ideas and has developed two major forums for dissemination of stem cell research and related issues, an annual meeting and an information-rich website. With the field of stem cell research and technologies rapidly advancing, discourse is more critical than ever to maintain high standards across the full spectrum of research. PMID- 17465795 TI - Stem cells & regenerative medicine: from molecular embryology to tissue engineering and therapeutics. PMID- 17465796 TI - VetCell Bioscience Ltd--regenerative medicine for the world of animal health. AB - VetCell Bioscience is a UK-based company focused on pioneering the use of regenerative medicine in the animal health market. VetCell was formed in partnership with the Royal Veterinary College and the Institute for Orthopaedic and Musculoskeletal Science to develop the use of cellular therapies to treat athletic injuries in horses. This ground-breaking work has been the springboard from which the Company has expanded into other areas of veterinary regenerative medicine. PMID- 17465797 TI - Heart failure and regenerative cardiology. PMID- 17465800 TI - Bioactive wound healing, bioaesthetics and biosurgery: three pillars of product development. Interview with Geoff MacKay. AB - Tissue regeneration specialist company Organogenesis Inc. was one of the first biotech companies formed. Incorporated in 1985, the company was originally a spin off from a research program at MIT. For the first 10-15 years, Organogenesis was heavily research based, but then gradually moved into development. The company's flagship product is Apligraf--a living, bilayered skin construct with two FDA approved indications: diabetic foot ulcers and venous leg ulcers. As Apligraf neared the market, it was necessary to 'graft' a manufacturing capability onto the company. As a consequence the company moved south from Massachusetts's cradle of biotechnology to Canton, MA, USA. Having experienced many of the highs and lows that characterize the biotech industry, the company is now consolidating its position as a center of expertise in commercializing living, cell-based products. The company has now built a sales, marketing and reimbursement team with the unique skill set to integrate novel technology into the US healthcare system. President & Chief Executive Officer Geoff MacKay takes great pride in the leading role that Organogenesis is playing in ushering in the field of tissue regeneration. Here, he discusses with Regenerative Medicine's Elisa Manzotti the 'three pillars' of the Organogenesis pipeline: bioactive wound healing, bioaesthetics and biosurgery. He focuses both on the rewards, and the trials and tribulations, of the commercialization of living cell-based technology. PMID- 17465801 TI - Potential application of embryonic stem cells in Parkinson's disease: drug screening and cell therapy. AB - Embryonic stem (ES) cells are genetically normal, continuous cell lines that can give rise to a variety of somatic cells in culture. These include the midbrain dopaminergic neurons, a major cell type lost in Parkinson's disease. With the promising outcome of mesencephalic fetal transplantation in some Parkinson's disease patients, the establishment of human ES cells has sparked much attention in both the scientific and general community regarding their potential as an alternative to aborted fetal tissue for cell replacement therapies. There is also great interest in developing the ES cell system as a platform for pharmaceutical and toxicological screening. Progress has been made in developing protocols for dopaminergic neuronal specification in ES cell development. Research to define the criteria for the 'right' category of therapeutic dopaminergic cells is underway. However, the promise of human ES cells rests largely on our ability to expand stem cells without genetic and epigenetic compromise, and to direct stem cell differentiation with absolute phenotypic fidelity. The delivery of these goals will require a much better understanding of the control of ES cell self renewal, proliferation and the commitment of differentiation. PMID- 17465802 TI - Human embryonic stem cell-derived cardiomyocytes: inducing strategies. AB - Cell-based therapy is emerging as an innovative approach for the treatment of degenerative diseases, and stem cells appear to be an ideal source of cells for this. In cardiology, in particular, human embryonic stem cell (hESC)-derived cardiomyocytes theoretically fulfill most, if not all, of the properties of an ideal donor cell, but several critical obstacles need to be overcome. Many research projects are focusing on set-up strategies for directing hESC differentiation toward the cardiac lineage. It is one of the main difficulties in the search to provide a valuable source of cells to effect regeneration of myocardial tissue in patients with severe heart failure. To date, there are no easy and efficient protocols for the induction of hESC differentiation toward the cardiac lineage. Discovering new molecules or tools capable of doing this is imperative. PMID- 17465803 TI - Notch signaling and its integration with other signaling mechanisms. AB - An improved understanding of stem cell differentiation is critical for progress in regenerative medicine. It is an emerging view that a relatively small number of intracellular signaling mechanisms play particularly important roles in differentiation control. As one may expect, these pathways are highly evolutionarily conserved, used in many tissues and iteratively during differentiation of a particular tissue. The Notch signaling system is one pathway meeting these criteria. In many cases, Notch signaling keeps stem/progenitor cells undifferentiated, although it can in some cellular contexts be instructive for differentiation toward a particular fate. Here, we review our current understanding of how Notch controls cellular differentiation in various organs and how Notch integrates with other major signaling pathways, primarily focusing on Notch signaling in mammals. Given the importance of Notch in many stem cell fate decisions, the possibility of experimentally manipulating Notch signaling opens up new avenues to control stem cell differentiation. PMID- 17465804 TI - New neurons for the injured brain: mechanisms of neuronal regeneration in adult teleost fish. AB - In contrast to mammals, teleost fish exhibit an enormous potential to continuously produce new neurons in many areas of the adult brain, and to regenerate neural tissue after brain injury. The regenerative capability of the teleost fish brain is based upon a series of well-orchestrated individual processes, including: elimination of damaged cells by apoptosis, removal of cellular debris by the action of microglia/macrophages, proliferation of endogenous neural precursor cells, radial glia-mediated migration of their progeny to the site of the lesion, neuronal differentiation, promotion of cellular survival, and integration of the new neurons into existing neural circuits. Combination of a well-defined cerebellar lesion paradigm with differential proteome analysis has demonstrated that identification of the multitude of proteins mediating the regenerative potential of the adult fish brain is feasible in the foreseeable future. A molecular understanding of brain regeneration in fish could help investigators to define novel strategies to stimulate endogenous neural precursor cells in the mammalian brain to undergo neurogenesis, thus forming the basis of a neuronal replacement therapy for brain injury or neurodegenerative diseases. PMID- 17465805 TI - Realizing the cardiac stem cell promise: a case for trophism. AB - Recent advances in stem cell biology have given rise the new field of cardiac regenerative medicine. Specifically, the development of cardiac stem cell science now offers the promise of novel cardiovascular therapies based on a dynamic body of basic and translational research. Importantly, the potential wide-spread clinical application of this technology will require that therapies be optimized for individuals with potential impairments in cardiac stem cell function. To this end, the previous experience of hematopoietic stem cell therapies can provide important guidance in the development and maturation of the young cardiac stem cell field. Parallel to the impact that exogenous growth factors have made in the field of hematopoietic therapies, the discovery and potential application of the factor(s) that govern cardiac regeneration may speed the progression of cardiac stem cell technology into an assessable and potent clinical therapy. PMID- 17465806 TI - Efficient transient genetic labeling of human CD34+ progenitor cells for in vivo application. AB - Genetic labeling of human hematopoietic progenitor cells (HPC) and their consecutive fate-mapping in vivo is an approach to answer intriguing questions in stem cell biology. We recently reported efficient transient genetic labeling of human CD34+ HPC with the truncated low-affinity nerve growth factor receptor (DeltaLNGFR) for in vivo application. Here we investigate whether HPC labeling with DeltaLNGFR affects lineage-specific cell differentiation, whether DeltaLNGFR expression is maintained during lineage-specific cell differentiation and which leukemia cell line might be an appropriate cell culture model for human CD34+ HPC. Human CD34+ peripheral blood stem cells and various leukemia cell lines were characterized by immunophenotyping. Cells were transfected using nucleofection. Hematopoietic differentiation was studied by colony-forming assays. DeltaLNGFR expression was assessed using reverse transcription-PCR, immunofluorescence and flow cytometry. Nucleofection was efficient and did not significantly reduce hematopoietic cell differentiation. Mature myeloid cells (CD66b+) derived from human CD34+ HPC and Mutz2 cells maintained DeltaLNGFR expression at a high percentage (70 +/- 2% and 58 +/- 2%, respectively). Mutz2 cells may serve as an in vitro model for human myeloid HPC. The method described herein has been adopted to Good Manufacturing Practices (GMP) guidelines and is ready for in vivo application. PMID- 17465807 TI - Regional and global protective effects of tissue kallikrein gene delivery to the peri-infarct myocardium. AB - BACKGROUND: The kallikrein-kinin system participates in the maintenance of the cardiovascular phenotype. We previously demonstrated that human tissue kallikrein gene (hTK) transfer promotes the healing of ischemic limbs. The present investigation aimed to test the original hypothesis that hTK delivery to the peri infarct myocardium would prevent post-ischemic heart failure. METHODS AND RESULTS: Myocardial infarction (MI) was induced in anesthetized mice by permanently occluding the left coronary descendant. hTK was delivered to the peri infarct myocardium via an adenoviral vector (Ad.hTK). Controls received Ad.Null or saline. Survival rate was similar among groups. Ad.hTK increased the number of circulating endothelial progenitor cells and promoted the growth of capillaries and arterioles in the peri-infarct myocardium. In addition, Ad.hTK increased the abundance of cardiac progenitor cells (CPCs) in the peri-infarct and suppressed the apoptotic death of peri-infarct cardiomyocytes in vivo and ex vivo. As a consequence of these beneficial effects, at 5 weeks from MI, hTK-transduced hearts were protected from post-MI ventricular dilatation and showed better systolic and diastolic functions. CONCLUSIONS: Ad.hTK benefits the neovascularization and viability of peri-infarct myocardium and increases CPC abundance, thereby decreasing ventricular dysfunction. Our study significantly adds to the knowledge of the protective effects of TK gene transfer on ischemic diseases and opens new avenues for the treatment of post-MI cardiac failure. PMID- 17465808 TI - Autologous adult rodent neural progenitor cell transplantation represents a feasible strategy to promote structural repair in the chronically injured spinal cord. AB - Adult neural progenitor cells (NPCs) represent an attractive source for cell based regenerative strategies in CNS disease. In animal models of spinal cord injury, syngenic adult NPCs, which were isolated from pooled post-mortem CNS tissue and co-transplanted together with fibroblasts, have been shown to promote substantial structural repair. The autologous transplantation of adult NPCs represents a major advantage compared with other sources of neural stem/progenitor cells. However, the feasibility of autologous NPC generation from a single biopsy in a relevant preclinical CNS disease model has yet to be demonstrated. To investigate this matter, adult Wistar rats underwent a cervical spinal cord lesion, which was followed by a minimal subventricular zone aspiration biopsy 2 days later. NPCs were isolated and propagated separately for each animal for the following 8 weeks. Thereafter, they were co-transplanted with simultaneously harvested skin fibroblasts in an autologous fashion into the cervical spinal cord lesion site. A total of 4 weeks later, graft survival, tissue replacement and axonal regeneration were assessed histologically. Animals receiving either allogenic NPCs combined with fibroblasts or autologous pure fibroblast grafts served as control groups. Within 8 weeks after the biopsy more than 3 million NPCs could be generated from a single aspiration biopsy, which displayed a differentiation pattern indistinguishable from syngenic NPC grafts. NPCs within autologous co-grafts readily survived, replaced cystic lesion defects completely and differentiated exclusively into glial phenotypes, thus paralleling previous findings with syngenic NPCs. The delayed transplantation 8 weeks after the spinal cord lesion elicited substantial axonal regeneration. These findings demonstrate that the therapeutic strategy to induce structural repair by transplanting adult autologous NPCs, after the successful propagation from a small brain biopsy into an acute CNS disease model, such as spinal cord injury, is feasible at the preclinical level. PMID- 17465809 TI - Autologous human-derived bone marrow cells exposed to a novel TGF-beta1 fusion protein for the treatment of critically sized tibial defect. AB - We report the first clinical case of transplantation of autologous bone marrow derived cells in vitro exposed to a novel recombinant human transforming growth factor (rhTGF)-beta1 fusion protein bearing a collagen-binding domain (rhTGF beta(1)-F2), dexamethasone (DEX) and beta-glycerophosphate (beta-GP). When such culture-expanded cells were loaded into porous ceramic scaffolds and transplanted into the bone defect of a 69-year-old man, they differentiated into bone tissue. Marrow cells were obtained from the iliac crest and cultured in collagen gels impregnated with rhTGF-beta1-F2. Cells were selected under serum-restricted conditions in rhTGF-beta(1)-F2-containing medium for 10 days, expanded in 20% serum for 22 days and osteoinduced for 3 additional days in DEX/beta-GP supplemented medium. We found that the cell number harvested from rhTGF-beta(1) F2-treated cultures was significantly higher (2.3- to 3-fold) than that from untreated cultures. rhTGF-beta(1)-F2 treatment also significantly increased alkaline phosphatase activity (2.2- to 5-fold) and osteocalcin synthesis, while calcium was only detected in rhTGF-beta(1)-F2-treated cells. Eight weeks after transplantation, most of the scaffold pores were filled with bone and marrow tissue. When we tested the same human cells treated in vitro in a rat model using diffusion chambers, there was subsequent development of cartilage and bone following the subcutaneous transplantation of rhTGF-beta(1)-F2-treated cells. This supports the suggestion that such cells were marrow-derived cells, with chondrogenic and osteogenic potential, whereas the untreated cells were not under the same conditions. The ability for differentiation into cartilage and bone tissues, combined with an extensive proliferation capacity, makes such a marrow derived stem cell population valuable to induce bone regeneration at skeletal defect sites. PMID- 17465811 TI - Patients and stem cells. PMID- 17465812 TI - ES Cell International & regenerative medicine. AB - ES Cell International believes that human embryonic stem cells offer a renewable source material for the elaboration of a multitude of different tissues for use in human cell replacement therapy. There are many challenges to the successful implementation of this vision including technical, regulatory, financial and ethical hurdles. However with demographic changes towards an aging population in the richer countries, leading to growing demands of ways to combat degenerative disease, science would be remiss not to explore every potential medical solution. PMID- 17465816 TI - Cell therapy--back on the up-curve. Interview with Paul Kemp by Elisa Manzotti. AB - Intercytex is an emerging healthcare company developing cell therapy products for the wound care and aesthetic medicine markets. The company, based in Cambridge, UK, commenced operations in 2000 and has raised over 31 million pounds in four private equity funding rounds. It is using its proprietary expertise in cell therapy to develop products that harness the innate ability of human cells to regenerate and repair the body. Here, founder and Chief Scientific Officer of Intercytex, Dr Paul Kemp, reflects on current status and future prospects for cell therapy with Regenerative Medicine's Elisa Manzotti. Dr Kemp was formerly Vice President of Research at Organogenesis, Inc. and has more than 17 years' experience in the commercial development of cell therapy. In this interview, Dr Kemp draws attention to a cycle that so often characterizes novel medical research: initial hype, a subsequent trough of disappointment and final emergence of viable technology. He explains how cell therapy is now emerging from the trough to become a rational, real and successful component of modern medicine. PMID- 17465817 TI - Adult neurogenesis and neurodegenerative disease. AB - Advances in stem cell biology of the adult brain and the discovery of adult neurogenesis have raised the hope that neurodegenerative disorders might ultimately become amenable to causal therapy. Stem cells contribute to cellular plasticity during the lifespan, and in some sense, brain development never ends. However, neurodegeneration is not just a lack of neuroregeneration, and cell genesis in the adult brain does not apparently lead to successful endogenous responses to neurodegeneration. The brain heals poorly; nevertheless, the onset, severity and progression of neurodegenerative disorders show large variation and can often be influenced by cognitive training and physical activity. Rather than providing endogenous repair, cellular plasticity, including adult neurogenesis might thus contribute to the 'cognitive reserve' that determines how well an organism can compensate for neurodegeneration. From this perspective, neurodegenerative disorders, such as Alzheimer's, Parkinson's, Lewy body and Huntington's diseases, might share a relevant biological principle that even links them to psychiatric disorders, like depression, which are not considered 'neurodegenerative' in a classical sense. However, the integration of neuroregenerative phenomena and most notably adult neurogenesis into the concepts of neurodegeneration is not without problems and remains speculative at present. Adult neurogenesis might be part of the physiological regenerative response and might thereby alter or alleviate symptoms, but it might also become impaired by the disease mechanism and thereby contribute to the symptoms of neurodegeneration. In any case, the extent to which effects on the level of cellular plasticity, be it degenerative or regenerative, are relevant functionally remains to be determined. The present review gives an overview of what is known about cell genesis and adult neurogenesis in neurodegenerative disorders and discusses how cellular plasticity might be part of concepts that integrate aspects of development and cellular plasticity into neurodegeneration. PMID- 17465818 TI - Corneal epithelial stem cells: characterization, culture and transplantation. AB - The epithelium covering the cornea at the front of the eye is maintained by stem cells located at its periphery, in a region known as the limbus. A lack or dysfunction of these so-called limbal stem cells (LSCs) results in the painful and blinding disease of LSC deficiency. In this review, current knowledge regarding the biology of these particular stem cells will be outlined, including recent advances that are enabling the gene expression analysis of these cells. The use of LSCs in therapeutic interventions for LSC deficiency will also be discussed, including the role for ex vivo expansion. In particular, the translation of basic science advances in LSC biology into therapeutic strategies will be highlighted. PMID- 17465819 TI - Regeneration of squamous epithelia from stem cells of cultured grafts. AB - The only cultured cell types extensively used for tissue regeneration are the keratinocyte and the chondrocyte. Cultured autologous keratinocytes derived from the epidermis have been used for many years to produce grafts that regenerate an epidermis over a full-thickness wound, such as a third-degree burn. But there have been many failures of engraftment, and in the absence of criteria for the quality of the cultures, the causes of failure cannot be analyzed. It has become clear that the essential feature of the graft is the presence of an adequate number of stem cells. This article describes the criteria for estimating that number. Advances in graft preparation, combining better preservation of stem cells with ease of application of the graft, are also described. These improvements have been applied to cultures of ocular limbal cells, which contain the keratinocyte stem cells of the corneal epithelium. Cultures meeting the criteria of stem cell number have been grafted to 116 patients suffering from chemical destruction of the limbus. The procedure has been highly successful in the alleviation of suffering and the restoration of vision. PMID- 17465820 TI - Reparative medicine: from tissue engineering to joint surface regeneration. AB - Biological regeneration is proving to be an increasingly attractive alternative/complement to prosthetic replacement of tissues and organs. In cell based therapeutic approaches, cells are manipulated in vitro and administered to patients as living and dynamic biological agents. In this review, we have focused on the regeneration of the injured joint surface to discuss novel issues that these new therapeutic agents raise and are difficult to address within the paradigms of traditional pharmacology. They include: determination of the mechanism of action and dose, evaluation of potency, safety and toxicity, as well as upscale, delivery and identification of proper indications. Finally, novel potential approaches are proposed in which resident progenitor cells are targeted to regenerate the damaged tissue. PMID- 17465821 TI - Oxygen, epigenetics and stem cell fate. AB - Coordinated interactions between the embryo/fetus and its environment are critical for proper development. In addition to acting as metabolic substrates for cellular homeostasis, basic physiological factors, such as oxygen tension, have a profound influence on developmental outcomes. Since the mammalian embryo resides in a physiologically hypoxic environment during gestation, understanding its responses to oxygen deprivation on a cellular level is critical. In this review, we analyze interactions between the hypoxia-inducible factor family of transcriptional regulators and epigenetic mechanisms governing chromatin structure. The ability of hypoxia-inducible factors to interact physically with histone deacetylase (HDAC) enzymes and modulate nuclear HDAC activities places them in the pivotal position of integrating physiological and epigenetic effectors. Multiple embryonic and extra-embryonic stem cell populations in mice and humans rely on this interaction--an important determinant of stem cell fate. Dissection of the pathways involved will provide novel insights into the metabolic as well as molecular determinants of the stem cell niches that allow self-renewal of progenitors in an undifferentiated state. PMID- 17465822 TI - Modeling human germ cell development with embryonic stem cells. AB - There has previously been no robust cell-based model for examining the genetic and epigenetic mechanisms of human germ cell formation. Human embryonic stem cells (hESCs) could potentially fill this need, as all cell types analyzed to date (including mature germ cells) can be identified by marker analysis during hESC differentiation. Furthermore, hESCs could also be used to differentiate mature female germ cells (oocytes) in culture as an alternate reprogramming cell for somatic cell nuclear transfer. However, to differentiate and isolate a functional germ cell from hESCs, the mechanisms that regulate germ cell formation need to be understood. The purpose of this review is to summarize the current understanding of the earliest events in human germ cell formation and to describe some of the known genetic pathways that regulate germ cell specification and development in the mouse. Finally, the current literature on the formation of germ cells from ESCs will be described. PMID- 17465823 TI - Culture of human embryonic stem cells and the extracellular matrix microenvironment. AB - Among the major obstacles impeding successful derivation and continuous culture of human embryonic stem cells (hESC) for therapeutic purposes, are the presence of feeder cells and feeder-conditioned media of animal origin. The risk of contamination with xenopathogens makes hESC cultured in this way unsafe for future use in regenerative medicine. A holy grail for investigators in the field will be to establish and maintain new hESC lines in completely feeder-free and serum-free defined conditions. Recently, propagation of hESC has become possible, using mammalian- or human-derived extracellular matrix (ECM) and conditioned medium from feeder cells. In addition, providing a three-dimensional ECM environment can even support the derivation of new hESC. In this review, we examine recent advances in the use and development of substrates suitable for the derivation and maintenance of hESC, and our current understanding of the effects of a three-dimensional ECM milieu on cellular behavior. PMID- 17465824 TI - Human embryonic stem cell derivation: from the IVF perspective to therapeutic applications. AB - Human embryonic stem cells (hESC) are capable of proliferating indefinitely in an undifferentiated state and are pluripotent, being able to differentiate into most cell types under the correct conditions. Since the establishment of the first hESC line in 1998, the hope has existed that these cells could constitute an unlimited cell source for replacement therapy in the treatment of various diseases and disabilities. However, there is opposition and concern within society towards hESC derivation. The purpose of this article is to introduce the medical and scientific issues surrounding hESC derivation for clinical use concerning the source for this research (human embryos donated from in vitro fertilization procedures), and the methodologies implicated in feeder-free, xeno free derivation that will allow potential clinical applications. PMID- 17465825 TI - Exploitation of adherent neural stem cells in basic and applied neurobiology. AB - Evidence for neurogenesis within the adult brain has challenged traditional views that this tissue is devoid of stem cell activity. This raises the possibility of introducing new cells through cell transplantation or stimulating endogenous neurogenesis as routes to treat disease and injury. Fetal and adult neural stem/progenitor cells can be isolated and expanded in vitro and might provide a cell source for such transplantations. Embryonic stem (ES) cells, which can generate any adult tissues, offer an alternative unlimited supply of neural tissue. We recently showed that both mouse and human ES cells can be converted to adherent neural stem (NS) cell lines [1] . Here we discuss the benefits of working with NS cell lines and how they might be exploited for studies of fundamental cellular processes, such as neuronal specification and differentiation. NS cells also serve as versatile models of disease processes, either through genetic manipulations or direct isolation from disease carriers and can be exploited in pharmaceutical drug screening. Longer term, NS cells offer an opportunity to rigorously test the efficacy of cell-based therapies and develop strategies for tissue engineering. PMID- 17465826 TI - Cardiac cells implanted within the outer aortic wall of rats generate measurable contractile force. AB - PURPOSE: To determine whether neonatal cardiomyocytes grafted into the aortic wall contract, develop pressure, and can be paced. METHODS AND RESULTS: Medium only (n = 9) or neonatal cardiomyocytes (n = 12, 5 x 10(6) cells each) were injected into the outer aortic wall in adult female Fischer rats. At 6 weeks after implantation, 11 out of 12 cardiomyocyte-treated aortas showed spontaneous rhythmic beating at the grafted site following excision of the heart. The spontaneous beating rate changed with pacing frequency. Five out of the 11 beating aortas had intra-aortic pressure generated by the spontaneously contracting cardiomyocytes. The pulse pressure generated by the grafted cardiomyocytes was 0.36 +/- 0.05 mmHg without pacing; during pacing it was 0.78 +/- 0.21 mmHg with systolic pressure up to 3.8 mmHg. Hematoxylin and eosin staining showed viable grafts in the outer wall of the cardiomyocyte-treated aortas in 12 out of 12 aortas. Neonatal cardiomyocytes in the graft matured with cross striations. Immunohistochemical staining of the aorta for sacromeric actin was positive in 12 out of 12 aortas. Staining of connexin 43 showed that some grafted cardiomyocytes formed gap junctions. The above examinations were negative in nine out of nine medium-treated aortas. CONCLUSION: The results show for the first time that cardiomyocytes engrafted into the foreign environment of an extracardiac vascular structure can be paced and generate measurable intravascular pressure. This study may serve as a useful model for studying the growth and response of the grafted cardiomyocytes to various stimuli in an extra cardiac environment in vivo. PMID- 17465827 TI - Telomerase promoter reprogramming and interaction with general transcription factors in the human mesenchymal stem cell. AB - The human adult mesenchymal stem cell (hMSC) does not express telomerase and has been shown to be the target for neoplastic transformation after transduction with hTERT. These findings lend support to the stem cell hypothesis of cancer development but by supplying hTERT, the molecular events required to upregulate hTERT expression in cancer development are missed. Therefore, the hMSC is ideal for the identification of molecular mechanisms regulating telomerase gene expression in stem cells. This study shows that the repression of hTERT expression in hMSC is chromatin based and that modifications of the chromatin environment lead to reactivation of telomerase gene expression. It is shown that repression of hTERT expression in hMSCs is due to promoter-specific histone hypoacetylation coupled with low Pol II and TFIIB trafficking. This repression is overcome by treatment with Trichostatin A (TSA), an HDAC inhibitor, concomitant with increases in promoter-specific histone acetylation and increases in Pol II and TFIIB tracking. hTR expression is also increased in TSA-treated hMSCs, concomitant with changes in Pol II and TFIIB dynamics. PMID- 17465830 TI - The UK Stem Cell Bank: a UK government-funded, international resource center for stem cell research. AB - The UK Stem Cell Bank is a UK Research Council-funded initiative that aims to provide ethically sourced and quality controlled stocks of cells for researchers and also establish seed stocks of cell lines for clinical trials. Whilst the Bank is prohibited from carrying out basic stem cell research (to avoid conflicts of interest) it is working to improve stem cell banking procedures including cryopreservation, characterization and quality control. The Bank also supports training activities and has provided the hub for the International Stem Cell Initiative, which includes 17 expert stem cell centers aiming to characterize a large number of human embryonic stem cell lines in a standardized way to improve our understanding of the characteristics of these cells. PMID- 17465831 TI - ReNeuron Group plc. AB - ReNeuron is a UK-based pioneer in stem cell research and development. The Company has leading edge, proprietary somatic stem cell technologies from which it is developing groundbreaking cell therapy products. ReNeuron's focus is on cell therapy treatments designed to reverse the effects of major diseases such as stroke, diabetes and diseases of the eye. PMID- 17465835 TI - The use of tissue engineering and stem cells in bladder regeneration. AB - Tissue engineering techniques for bladder regeneration have been applied successfully for many years in a variety of in vitro and in vivo models. But despite these rapid advances, to date, none of the tissue-engineered constructs could be used in human models due to inconsistent results of the described techniques in animal models. Three factors have been identified to influence the regeneration process: identification of the ideal scaffold, appropriate cell population for seeding and the optimal regeneration conditions necessary. Identifying the role of each component will help to unlock the complex regeneration mechanisms required to achieve consistent, reliable results that will allow transition of the technology into clinical practice. This review will discuss the role and applicability of the each factor and provide a future prospective on tissue engineering techniques for bladder regeneration. PMID- 17465832 TI - Large animal models are critical for rationally advancing regenerative therapies. PMID- 17465836 TI - The role of hyaluronic acid in hemopoietic stem cell biology. AB - The adult mammalian hemopoietic system maintains an extraordinarily large, yet well regulated supply of mature blood cells within the circulation throughout life. The system is capable of rapid recovery and compensation following injury, environmental stress or as a result of genetic disease such as the hemoglobinopathies. Despite the vast amount of research conducted there is still an incomplete understanding of hemopoietic regulation. Nevertheless, it is evident from transplantation studies that ongoing blood cell production is absolutely dependent upon hemopoietic stem cells (HSCs). These rare and potent cells have the capacity for extensive proliferation and the ability to differentiate into all blood cell types. An understanding of HSC regulation is fundamental to understanding hemopoiesis. There is now considerable evidence to demonstrate that in vivo, HSCs are located within defined anatomical sites or niches within the bone marrow. Regulation of HSC fate is mediated by both cell autonomous mechanisms and extrinsic cues resulting from interactions between cells and extracellular components within the niche. This review focuses on the role of hyaluronic acid, a component of the HSC niche and moreover a HSC associated glycosaminoglycan, in hemopoiesis and specifically HSC regulation. It is now evident that hyaluronic acid not only provides a physical scaffold or support within the marrow to facilitate localization and retention of HSCs to the stem cell niche but moreover, through ligation with its counter-receptors is able to directly affect the cellular functions of HSCs. PMID- 17465837 TI - Multifunctional cell-instructive materials for tissue regeneration. AB - Current approaches in tissue engineering and regenerative medicine have focused on controlling the presentation of various factors that influence cellular behavior and tissue formation. Numerous biomaterials have been utilized as sites for new tissue growth by migrating or transplanted cells, nanoscale control of cellular behavior through the presentation of specific peptide sequences, and depots for growth factor release. More recently, the development of bioresponsive materials has emerged as a promising approach to cede control of temporal macromolecule presentation and material degradation to invading cell populations. Biomaterials now have the potential of possessing multiple functions in the process of tissue regeneration. This review summarizes some of the recent advances in the use of multifunctional biomaterials in the arena of tissue engineering. Specifically, the potential of various materials is described as it pertains to the control of cellular behavior, integration of engineered materials with host or transplanted tissue, and inductive factor presentation. PMID- 17465838 TI - Biological and mechanical enhancement of impacted allograft seeded with human bone marrow stromal cells: potential clinical role in impaction bone grafting. AB - With the demographics of an aging population the incidence of revision surgery is rapidly increasing. Clinical imperatives to augment skeletal tissue loss have brought mesenchymal stem cells to the fore in combination with the emerging discipline of tissue engineering. Impaction bone grafting for revision hip surgery is a recognized technique to reconstitute bone, the success of which relies on a combination of mechanical and biological factors. The use of morsellized allograft is currently the accepted clinical standard providing a good mechanical scaffold with little osteoinductive biological potential. We propose that applying the principles of a tissue engineering paradigm, the combination of human bone marrow stromal cells (hBMSCs) with allograft to produce a living composite, offers a biological and mechanical advantage over the current gold standard of allograft alone. This study demonstrates that hBMSCs combined with allograft can withstand the forces equivalent to a standard femoral impaction and continue to differentiate and proliferate along the bony lineage. In addition, the living composite provides a biomechanical advantage, with increased interparticulate cohesion and shear strength when compared with allograft alone. PMID- 17465843 TI - Tissue engineering through mesenchymal stem cells: role of the Genostem Consortium. PMID- 17465839 TI - Transplantation of human embryonic stem cell-derived oligodendrocyte progenitors into rat spinal cord injuries does not cause harm. AB - Demyelination contributes to loss of function following spinal cord injury. We have shown previously that transplantation of human embryonic stem cell-derived oligodendrocyte progenitors into adult rat 200 kD contusive spinal cord injury sites enhances remyelination and promotes recovery of motor function. Previous studies using oligodendrocyte lineage cells have noted a correlation between the presence of demyelinating pathology and the survival and migration rate of the transplanted cells. The present study compared the survival and migration of human embryonic stem cell-derived oligodendrocyte progenitors injected 7 days after a 200 or 50 kD contusive spinal cord injury, as well as the locomotor outcome of transplantation. Our findings indicate that a 200 kD spinal cord injury induces extensive demyelination, whereas a 50 kD spinal cord injury induces no detectable demyelination. Cells transplanted into the 200 kD injury group survived, migrated, and resulted in robust remyelination, replicating our previous studies. In contrast, cells transplanted into the 50 kD injury group survived, exhibited limited migration, and failed to induce remyelination as demyelination in this injury group was absent. Animals that received a 50 kD injury displayed only a transient decline in locomotor function as a result of the injury. Importantly, human embryonic stem cell-derived oligodendrocyte progenitor transplants into the 50 kD injury group did not cause a further decline in locomotion. Our studies highlight the importance of a demyelinating pathology as a prerequisite for the function of transplanted myelinogenic cells. In addition, our results indicate that transplantation of human embryonic stem cell-derived oligodendrocyte progenitor cells into the injured spinal cord is not associated with a decline in locomotor function. PMID- 17465844 TI - The concept of mesenchymal stem cells. AB - In this chapter we examine whether criteria usually defining adult tissue stem cells apply to mesenchymal stem cells (MSCs) that give rise to cells of the skeletal connective tissues. MSCs appear to constitute a heterogeneous population of undifferentiated and committed, lineage-primed cells, capable of: homing upon engraftment to a number of growth microenvironments, extensive proliferation, producing large numbers of differentiated progeny, and functional tissue repair after injury. In addition, MSCs are extensively distributed throughout tissues, and bone marrow MSCs provide the stromal component of the niche of hematopoietic stem cells. The capacity of apparently differentiated mesenchymal cells to shift their differentiation pathway with changing microenvironmental conditions (known as differentiation plasticity) may be due to de-differentiation and reprogramming in MSCs. Because they present several features setting them apart from other stem cells, MSCs may constitute another paradigm for stem cell systems, where self renewal and hierarchy are no longer essential, but where plasticity is the major characteristic. PMID- 17465845 TI - Proteomics of primary mesenchymal stem cells. AB - Tissue and functional regeneration takes place in the body at various stages throughout life. However, bone, cartilage, tendons, blood vessels and cardiac muscle have a limited capacity for self repair and, after injury or disease, the regenerative ability of these adult tissues is often insufficient and leads to nonfunctional scar tissue. In this context, mesenchymal stem cells, which are adult multipotential progenitors of mesoderm cells (osteoblasts, chondrocytes, adipocytes and stroma cells), represent a major hope for tissue-engineered replacement and regenerative medicine. Furthermore, the autologous use of these cells prevents immunological responses against new tissues and the risks of disease transmission from donors, which are both common problems of organ transplantation. While the existence of mesenchymal stem cells is undisputed, many questions remain regarding their self-renewal and capacity to differentiate, their homogenous nature as a cell population throughout the body and their true potential in regenerative medicine. In this article, the proteomics studies carried out to characterize mesenchymal stem cells and to help understand their physiology are reviewed. PMID- 17465846 TI - 3D scaffolds for bone marrow stem cell support in bone repair. AB - Bone tissue repair is one of the major concerns of regenerative medicine. The current need for tissue replacements has necessitated the development of a new science termed 'bone tissue engineering'. The basic organization of bone tissue requires the design and fabrication of a porous 3D structure or 'scaffold' to contain the bone-forming cells. This scaffold should be formulated from biocompatible, osteoconductive materials that are not immunoreactive. 3D scaffolds provide the necessary support for cells to proliferate and maintain their capacity to differentiate and scaffolds containing bone marrow-derived osteoprogenitors can be employed within implants to enhance bone repair. The complex construct is intended to mimic the native in vivo microenvironment and this demands construction of bioactive scaffolds that are also capable of supporting vascularization as well as cell proliferation and osteogenic differentiation. 3D bioactive scaffolds containing committed osteoprogenitors can provide a promising surgical tool for bone tissue engineering directed at orthopedic and cranio-maxillofacial clinical applications. PMID- 17465847 TI - Engineered mesenchymal stem cells for cartilage repair. AB - Healthy cartilage is a highly robust tissue, and is resilient against the stringent mechanical and biological constraints imposed upon it. Cartilage defects are common features of joint diseases, but current treatments can rarely restore the full function of native cartilage. Recent studies have provided new perspectives for cartilage engineering using mesenchymal stem cells (MSCs). However, the sequential events occurring during chondrogenesis must be fully understood before we are able to reproduce faithfully the complex molecular events that lead to MSC differentiation and long-term maintenance of cartilage characteristics. Here, we focus on the potential of MSCs to repair cartilage with an emphasis on the factors that are known to be required in inducing chondrogenesis. PMID- 17465848 TI - Osteogenic differentiation of human marrow-derived mesenchymal stem cells. AB - Bone marrow-derived mesenchymal stem cells (MSCs) are adherent cells that differentiate into chondroblasts, osteoblasts and adipocytes. In this short review, we summarize the molecular mechanisms that are known to control osteoblast differentiation and osteogenic potential of MSCs in vitro. We discuss the advances made in gene-based therapy to promote osteogenic differentiation of MSCs and the perspectives for an optimal use of MSCs for bone tissue regeneration or repair. One important challenge at the present time is to identify factors and pathways that promote osteogenic commitment of MSCs in order to use MSCs with functional potential for optimal bone repair in humans. In this context, genomic and proteomic analyses may help to identify molecules that could be used to promote osteogenic differentiation of human MSCs. In the future this may lead to selective therapeutic strategies for tissue engineering application in bone regeneration and repair in humans. PMID- 17465849 TI - Applications of gene therapy and adult stem cells in bone bioengineering. AB - Bone tissue engineering is an emerging field, that could become a main therapeutic strategy in orthopedics in coming years. While bone has regenerative abilities that enable the self repair and regeneration of fractures, there are extreme situations in which the extent of bone loss is too large for complete regeneration to occur. In order to achieve bone regeneration, osteogenic genes (mainly from the bone morphogenetic protein family) can be delivered either directly into the target tissue, or by using adult stem cells, which are later implanted into the target site. Engineered adult stem cells combined with biodegradable polymeric scaffolds can be implanted into target sites, with or without ex vivo culture period. Several important factors influence the success of bone engineering approaches including: choice of cell and scaffold, the vector used in order to deliver the osteogenic gene, and the osteogenic gene itself. Cutting-edge imaging technologies, bioinformatics-based analysis of gene expression and exogenous regulation of transgene expression are among the tools that are being used to optimize and control bone formation in vivo. In this review we have attempted to provide an overview of the main factors that should be considered when utilizing adult stem cells and gene therapy strategies to regenerate bone defects or to promote new bone formation in vivo. PMID- 17465850 TI - Tendon and ligament engineering: from cell biology to in vivo application. AB - Tendons and ligaments are related connective tissues that join muscle to bone and bone to bone, respectively. Tendon and ligament injuries are widely distributed clinical problems in society and while healing of such disorders can occur, the original biological properties of the tissue do not return to normal. In this review, recent work on tendon and ligament development and the use of growth factors for successful cellular therapy of tendon and ligament disorders are discussed. In addition, anti-inflammatory concepts for the treatment of tendon and ligament injuries and recent developments in stem cell engineering for tendon and ligament tissues are examined. Lastly, gene transfer strategies for therapeutic applications to heal tendon and ligament disorders are reviewed. PMID- 17465851 TI - Engineering stem cells for therapy. AB - The differentiation of a stem cell is dependent on the environmental cues that it receives and can be modulated by the expression of different master regulators or by secreted factors or inducers. The use of genetically modified stem cells to express the required factors can direct differentiation along the requisite pathway. This approach to the engineering of stem cells is important, as control of the pluripotentiality of stem cells is necessary in order to avoid unwanted growth, migration or differentiation to nontarget tissues. The authors provide an overview of the stem cell engineering field, highlighting challenges and solutions, and focusing on recent developments in therapeutic applications in areas such as autoimmunity, CNS lesions, bone and joint diseases, cancer and myocardial infarction. PMID- 17465852 TI - Mesenchymal stem cells in bone and cartilage repair: current status. AB - The progression of rheumatoid pathologies, degenerative diseases, traumatologies, and their cortege of increasing medical, social and economical needs, has mandated the development of tissue repair and engineering technologies in orthopedic medicine. Mesenchymal stem cells (MSCs) are multipotent cells that can be extracted from large and relatively easily accessible compartments of the body, especially the bone marrow, and such cells are able to differentiate into adipogenic, chondrogenic and osteogenic precursors. The concept of using MSCs to repair tissues has progressively evolved, and the goal of cell-mediated therapy is to prolong the natural physiological abilities of healing, or substitute them, when these are lacking, failing or progressing too slowly. In recent years, the first clinical trials on the utility of MSCs, with or without scaffolds and/or growth factors, have been initiated. In this review, the authors focus on findings from preclinical research, clinical trials and case reports involving bone and cartilage repairs. New perspectives are considered regarding uses of cell types, cell delivery approaches and growth factors. They also consider the stringent conditions, constraints and considerations necessary to take cell mediated therapy from bench to bedside. PMID- 17465853 TI - Splanchnic regulation of glucose production. AB - The liver plays a key role for the maintenance of blood glucose homeostasis under widely changing physiological conditions. In the overnight fasted state, breakdown of hepatic glycogen and synthesis of glucose from lactate, amino acids, glycerol, and pyruvate contribute about equally to hepatic glucose production. Postprandial glucose uptake by the liver is determined by the size of the glucose load reaching the liver, the rise in insulin concentration, and the route of glucose delivery. Hepatic glycogen stores are depleted within 36 to 48 hours of fasting, but gluconeogenesis continues to provide glucose for tissues with an obligatory glucose requirement. Glucose output from the liver increases during exercise; during short-term intensive exertion, hepatic glycogenolysis is the primary source of extra glucose for skeletal muscle, and during prolonged exercise, hepatic gluconeogenesis becomes gradually more important in keeping with falling insulin and rising glucagon levels. Type 1 diabetes is accompanied by diminished hepatic glycogen stores, augmented gluconeogenesis, and increased basal hepatic glucose production in proportion to the severity of the diabetic state. The hyperglycemia of type 2 diabetes is in part caused by an overproduction of glucose from the liver that is secondary to accelerated gluconeogenesis. PMID- 17465854 TI - Energy metabolism during human pregnancy. AB - This review summarizes information regarding how human energy metabolism is affected by pregnancy, and current estimates of energy requirements during pregnancy are presented. Such estimates can be calculated using either increases in basal metabolic rate (BMR) or increases in total energy expenditure (TEE). The two modes of calculation give similar results for a complete pregnancy but different distributions of energy requirements in the three trimesters. Recent information is presented regarding the effect of pregnancy on BMR, TEE, diet induced thermogenesis, and physical activity. The validity of energy intake (EI) data recently assessed in well-nourished pregnant women was evaluated using information regarding energy metabolism during pregnancy. The results show that underreporting of EI is common during pregnancy and indicate that additional longitudinal studies, taking the total energy budget during pregnancy into account, are needed to satisfactorily define energy requirements during the three trimesters of gestation. PMID- 17465855 TI - Metabolic regulation and function of glutathione peroxidase-1. AB - Glutathione peroxidase-1 (GPX1) represents the first identified mammalian selenoprotein, and our understanding in the metabolic regulation and function of this abundant selenoenzyme has greatly advanced during the past decade. Selenocysteine insertion sequence-associating factors, adenosine, and Abl and Arg tyrosine kinases are potent, Se-independent regulators of GPX1 gene, protein, and activity. Overwhelming evidences have been generated using the GPX1 knockout and transgenic mice for the in vivo protective role of GPX1 in coping with oxidative injury and death mediated by reactive oxygen species. However, GPX1 exerts an intriguing dual role in reactive nitrogen species (RNS)-related oxidative stress. Strikingly, knockout of GPX1 rendered mice resistant to toxicities of drugs including acetaminophen and kainic acid, known as RNS inducers. Intracellular and tissue levels of GPX1 activity affect apoptotic signaling pathway, protein kinase phosphorylation, and oxidant-mediated activation of NFkappaB. Data are accumulating to link alteration or abnormality of GPX1 expression to etiology of cancer, cardiovascular disease, neurodegeneration, autoimmune disease, and diabetes. Future research should focus on the mechanism of GPX1 in the pathogeneses and potential applications of GPX1 manipulation in the treatment of these disorders. PMID- 17465856 TI - Epigenetic epidemiology of the developmental origins hypothesis. AB - Extensive human epidemiologic and animal model data indicate that during critical periods of prenatal and postnatal mammalian development, nutrition and other environmental stimuli influence developmental pathways and thereby induce permanent changes in metabolism and chronic disease susceptibility. The biologic mechanisms underlying this "developmental origins hypothesis" are poorly understood. This review focuses on the likely involvement of epigenetic mechanisms in the developmental origins of health and disease (DOHaD). We describe permanent effects of transient environmental influences on the developmental establishment of epigenetic gene regulation and evidence linking epigenetic dysregulation with human disease. We propose a definition of "epigenetic epidemiology" and delineate how this emerging field provides a basis from which to explore the role of epigenetic mechanisms in DOHaD. We suggest strategies for future human epidemiologic studies to identify causal associations between early exposures, long-term changes in epigenetic regulation, and disease, which may ultimately enable specific early-life interventions to improve human health. PMID- 17465857 TI - Climate change, the butterfly effect, and the thyroid. PMID- 17465858 TI - Molecular cytogenetic profiles of novel and established human anaplastic thyroid carcinoma models. AB - In this study we present two novel anaplastic thyroid carcinoma (ATC) lines (HTh 104 and HTh 112) and further characterize six frequently used ATC lines (HTh 7, HTh 74, HTh 83, C 643, KAT-4, and SW 1736). Three of the lines carried a heterozygous BRAF mutation V600E, which is in line with reports of BRAF mutations in primary ATC and papillary thyroid cancer. Several nonrandom breakpoints were identified by spectral karyotyping (SKY) and G-banding in these lines including the novel 1p36 and 17q24-25 as well as 3p21-22 and 15q26 that are also implicated in well-differentiated thyroid cancers. Comparative genomic hybridization showed frequent gain of 20q, including the UBCH10 gene in 20q13.12, which was further confirmed by array-comparative genomic hybridization and fluorescence in situ hybridization analyses. Our results concur with previous studies in both primary tumors and cell lines, indicating that gain of chromosome 20 is important in the pathogenesis of ATC and/or progression of differentiated thyroid cancers to ATC. PMID- 17465859 TI - Tandem mass spectrometry improves the accuracy of free thyroxine measurements during pregnancy. AB - OBJECTIVE: Pregnancy is a time of rapidly changing demands on the thyroid axis, and knowledge of thyroid hormone levels, especially during the first trimester, is important for ensuring maternal and fetal health. The thyroid hormone assays currently in use become more inaccurate at extremes of binding protein concentrations and when heterophilic antibodies are present. Pregnancy is characterized by both these conditions, making accurate determination of free thyroid hormone levels by conventional direct analog immunoassay methods difficult. The objective of this study was to characterize the performance of a novel tandem mass spectrometric assay for free thyroxine during the physiologic conditions of pregnancy. DESIGN: Healthy women without a history of thyroid abnormalities were recruited from the obstetrics and gynecology and endocrinology clinics of a university medical center and their thyroid status was monitored. Free thyroxine levels were assessed by both immunoassay and tandem mass spectrometry during the course of their pregnancy. Serum thyrotropin levels were also measured. The distributions of free thyroid concentrations obtained by the two assays were compared. MAIN OUTCOME: The tandem mass spectrometry and immunoassay values did not correlate well with each other. However, tandem mass spectrometry values correlated well with the current gold standard equilibrium dialysis values. Moreover, the good agreement between equilibrium dialysis and tandem mass spectrometry was maintained across all weeks of gestation. CONCLUSIONS: We conclude that tandem mass spectrometry has a superior performance to immunoassay for the measurement of free thyroxine during pregnancy. Furthermore, it is ideally suited to generating trimester-specific reference intervals for free thyroxine levels. Future studies will determine if it is a better assay to use in most clinical circumstances. PMID- 17465860 TI - Aspects of treatment of subclinical hypothyroidism. PMID- 17465861 TI - Needle-free delivery of lidocaine for reducing the pain associated with the fine needle aspiration biopsy of thyroid nodules: time-saving and efficacious procedure. AB - OBJECTIVE: Fine-needle aspiration biopsy (FNAB) is a mandatory procedure in evaluation of thyroid nodules. However, it is sometimes perceived as a painful procedure by the patients. Efficacy of the needle-free injection of local anesthesia for reducing the pain associated with other cutaneous procedures that involve needle insertion was previously reported. In this double-blind, placebo controlled clinical trial, we evaluated the effectiveness of a needle-free injection of lidocaine in achieving satisfactory pain control in patients undergoing FNAB of thyroid nodules. DESIGN: Patients were allocated to receive either lidocaine administered by needle-free injection system (n = 55) or placebo (isotonic saline) (n = 52) 2-3 minutes before FNAB. A series of four aspirations of each nodule was performed. The patients rated pain associated with the procedure according to a 100-mm visual analog scale (VAS), an 11-point numeric rating scale (NRS), and a four-category verbal rating scale (VRS). MAIN OUTCOME MEASURE: The two groups studied were similar with respect to age, sex, thyroid volume, nodule size, and nodule site. When the effectiveness of lidocaine was compared with that of placebo, the mean VAS score was 11.4 +/- 13.6 mm versus 38.2 +/- 35.5 mm (p < 0.0001) and the mean NRS score was 1.4 +/- 1.5 points versus 3.9 +/- 2.6 points (p < 0.0001), respectively. The absolute numbers according to VRS score in each group was also significantly different (p < 0.0001). The percentage of patients with "no pain" or "mild pain" in the lidocaine group (90.9%) was significantly higher than that in the placebo group (44.2%) (p < 0.0001). Less than 10% of the patients in lidocaine group experienced moderate pain and none experienced severe pain. No adverse treatment related effects were observed. CONCLUSIONS: To our knowledge, this is the first study demonstrating that the needle-free delivery of lidocaine is an effective, useful, and noninvasive method of providing local anesthesia for the FNAB of thyroid nodules. PMID- 17465862 TI - Effects of either LT4 monotherapy or LT4/LT3 combined therapy in patients totally thyroidectomized for thyroid cancer. AB - After total thyroidectomy all thyroid cancer patients require lifelong treatment with thyroid hormones; the treatment of choice is synthetic levothyroxine (LT4). The question of whether these patients might benefit from the combined LT4 and liothyronine (LT3) treatment has been addressed with conflicting conclusions. The aim of the present study was to compare the effects of combined low LT4/LT3 molar ratio therapy versus LT4 monotherapy on various target organs and tissues in patients thyroidectomized for thyroid cancer. Urine collection (24 hour), a fasting blood sample for laboratory examinations, thyroid function clinical score, and cardiovascular, neurological, and neuropsychological evaluations were obtained. Clinical parameters and peripheral markers of thyroid function were measured during the two different treatment regimens in 20 patients. Mean serum aspartate aminotransferase, alanine aminotransferase, sex hormone binding globulin, and osteocalcin values were significantly higher during the combined treatment. No significant differences in the clinical score, the systolic and diastolic performance, and the neurological and neuropsychological evaluations were observed between the two treatment regimens. Moreover, no alteration due to subclinical hyperthyroidism or to the fluctuations in serum T3 concentrations during the combined therapy was observed. In conclusion, we found no evidence that combined therapy with a low LT4/LT3 molar ratio resulted in improved well being and cognitive function or in increased thyroid hormone action on peripheral tissues in respect to LT4 monotherapy. Until future large, blind, randomized, and controlled trials prove otherwise, LT4 should remain the standard treatment for thyroid cancer patients. PMID- 17465863 TI - Hyperparathyroidism in patients over 80: clinical characteristics and their ability to undergo outpatient parathyroidectomy. AB - OBJECTIVE: Parathyroid surgery in elderly patients is commonly delayed due to perceived high operative risk. We evaluated the presentation of patients over 80 with primary hyperparathyroidism (PHPT) and their ability to tolerate outpatient, minimally invasive parathyroidectomy. METHODS: We conducted a prospective cohort study of 150 consecutive patients over the age of 80 for the treatment of sporadic, nonfamilial PHPT. Presenting symptoms were compiled by questionnaire and compared between male and female (32 males, 118 females) and to 2600 patients under the age of 80. All patients underwent minimally invasive parathyroidectomy and were discharged from the recovery room. Operative findings and outcomes were assessed. RESULTS: 97% reported at least one symptom--the majority had five or more. The most common symptoms were fatigue, hypertension, and memory problems (occurring in 62%, 62%, and 57% of patients, respectively). Symptoms were similar between men and women, with the exception of bone pain being twice as common in women and kidney stones being twice as common in men (both p < 0.05). Preoperative calcium and parathyroid hormone (PTH) levels as well as the frequency of each symptom closely paralleled those of patients less than 80 years old with no significant differences. Average operative time was 18 +/- 5 minutes with discharge averaging 1.9 +/- 0.2 hours later. The incidence of single adenoma, double adenoma, or hyperplasia was identical to patients less than 80 (p = NS). Two patients required rehospitalization within 30 days of the procedure (congestive heart failure and pulmonary embolism), neither one for hypocalcemia. There were no deaths, and the cure rate was 99.3%. CONCLUSION: PHPT is similar symptomatically, biochemically, and histopathologically between patients > 80 years old and younger patients. Modern techniques allow for small incisions, quick operative times, outpatient discharge, and uneventful recovery. Patients over 80 years old tolerate outpatient parathyroidectomy without event. PMID- 17465864 TI - Enhanced scintigraphic visualization of thyroglossal duct remnant during hypothyroidism after total thyroidectomy: prevalence and clinical implication in patients with differentiated thyroid cancer. AB - OBJECTIVE: The exact prevalence and clinical significance of ectopic thyroid or thyroglossal duct remnant (TGDR) in the general population have not yet been fully determined despite numerous case reports. This study was prepared to assess the prevalence of TGDR in asymptomatic subjects during hypothyroidism after a total thyroidectomy for differentiated thyroid cancer (DTC) and to clarify the clinical implication. DESIGN: Tc-99m pertechnetate scintigraphy (Tc-scan) of the head and neck before radioiodine ablation therapy and whole-body and pinhole I 131 scintigraphy (I-scan) after ablation therapy were reviewed for 131 consecutive DTC patients with hypothyroidism after a total thyroidectomy. MAIN OUTCOME: Forty-four among the 131 patients (33.6%) revealed an unexpected linear or focal radioactivity at the anterior midline of the neck, suggesting the presence of TGDR. The Tc-scan and pinhole I-scan were concordant in all cases of abnormal midline neck uptake, although the planar I-scan failed to delineate TGDR due to prominent photon scattering in most cases. Preoperative enhanced neck computed tomography scan was performed in 49 patients and showed no evidence of thyroid glandular tissue separated from thyroid gland in midline of the anterior neck except 1 case. The success rate after radioiodine ablation did not differ significantly between the positive and negative TGDR patients. CONCLUSIONS: TGDR can be frequently observed in scintigraphy of hypothyroid subjects after a thyroidectomy, even when clinically unexpected. Therefore, care should be taken not to confuse the tracer uptake by TGDR with metastatic foci in I- and Tc-scans of patients with hypothyroidism after a thyroidectomy for DTC. PMID- 17465865 TI - Percutaneous ethanol injection therapy for locally recurrent papillary thyroid carcinoma. AB - OBJECTIVE: Percutaneous ethanol injection therapy (PEIT) has been used to treat various human neoplasms. We evaluated the local therapeutic effect of PEIT in recurrent thyroid cancers. DESIGN: From October 2002 to September 2005, we performed ultrasound-guided PEIT on 24 recurrent lesions (eight in thyroid beds and 16 in neck nodes) of 16 papillary thyroid carcinoma patients. Ethanol was injected at 3-month intervals under sonographic guidance. MAIN OUTCOME: All patients tolerated PEIT well with only mild local pain, although one patient complained of transient hoarseness after the procedure. The median diameter of lesions was significantly reduced, from 9.9 mm (range, 5.5-25.0 mm) to 5.3 mm (range, 0.0-17.0 mm) by PEIT. Four recurrent lesions disappeared sonographically. CONCLUSION: PEIT may be an alternative treatment option for locally recurrent thyroid carcinomas in properly selected patients. However, a prospective, long term follow-up study is necessary to determine the effect of PEIT on patient survival and tumor recurrence. PMID- 17465866 TI - Sunitinib induces hypothyroidism in advanced cancer patients and may inhibit thyroid peroxidase activity. AB - OBJECTIVE: Sunitinib is a novel tyrosine kinase inhibitor with antitumor and antiangiogenic effects. An observed higher than expected rate of hypothyroidism in sunitinib-treated patients prompted assessment of the incidence of hypothyroidism. DESIGN: Patients taking sunitinib had their thyroid function tests (TFTs) assessed via chart review. To explore potential effects on the thyroid, we examined the antiperoxidase activity of sunitinib by in vitro testing its effect on guaiacol oxidation and protein iodination by lactoperoxidase. MAIN OUTCOME: Of the 89 patients who took sunitinib, 49 patients were excluded from analysis for several reasons. Of the remaining 40 patients, 21 (53%, 24% of the original 89) developed elevated thyrotropin (TSH) after a median of 5 months (range 1-36 months). Median TSH was 21.4 mU/L (range 4.6-174 mU/L). In vitro, sunitinib had antiperoxidase activity that was about one-fourth the potency of propylthiouracil. CONCLUSIONS: Of the 40 patients who had TFTs assessed after starting sunitinib, 53% developed elevated TSH. We recommend that all patients treated with sunitinib be monitored for hypothyroidism. The mechanism of the antithyroid effect appears to be inhibition of peroxidase activity. Further research is needed to confirm the mechanism by which sunitinib induces hypothyroidism. PMID- 17465867 TI - Determinants of liver damage associated with intravenous methylprednisolone pulse therapy in Graves' ophthalmopathy. AB - BACKGROUND: Intravenous methylprednisolone pulses (IVMP) are more efficacious and better tolerated than oral prednisone in Graves' ophthalmopathy (GO) patients. However, acute and severe liver damage has been reported in sporadic cases during IVMP, resulting in fatal acute liver failure in four patients so far. The mechanism causing the liver damage is incompletely understood. DESIGN: We performed a prospective observational study in 13 patients with dysthyroid optic neuropathy (group A) and in 14 patients with moderately severe GO (group B) who were treated with high-dose (group A) or low-dose (group B) IVMP; cumulative steroid doses were 8.45 g in group A and 4.5 g in group B, and follow-up time was 24 weeks. MAIN OUTCOME: Slight increases in serum aminotransferases (in alanine aminotransferase [ALAT] more than in aspartate aminotransferase [ASAT]) were observed, in seven patients exceeding the upper normal limit of 40 U/L. These changes were more prominent in group A than in group B as was also evident from a decrease in ASAT/ALAT ratio in group A but not in group B. Changes in serum aminotransferases occurred especially in the first 6 weeks of IVMP, becoming smaller thereafter with the decrease in steroid dosage. Pretreatment liver steatosis or diabetes were not related to liver damage, but preexistent viral hepatitis was. CONCLUSION: IVMP in GO patients causes dose-dependent liver damage by a direct toxic effect of glucocorticoids on hepatocytes. Nevertheless, IVMP seems to be pretty safe if cumulative doses exceeding 8 g are avoided and liver function is checked before and at regular intervals during pulse therapy. PMID- 17465868 TI - Radioiodine uptake in non-lactating mammary glands: evidence for a causative role of hyperprolactinemia. AB - CONTEXT: Radioiodine uptake is rarely observed in normal non-lactating breast tissue. Investigation of the in vivo regulation of iodide uptake in breast tissue may be useful for the induction of radioiodine uptake in breast cancer tissue for diagnostic and therapeutic purposes. CASE REPORTS: We report the cases of two post-menopausal women who underwent radioiodine therapy for papillary thyroid carcinoma and in whom breast uptake of radioiodine on post-therapy whole body scan (WBS) was observed. METHODS AND RESULTS: In both patients, elevated serum prolactin levels (123 ng/mL in patient 1 and 48 ng/mL in patient 2) were documented at the time when radioiodine uptake in the breast was observed. The hyperprolactinemia was due to prolonged treatment with the anti-dopaminergic neuroleptic risperidone in Case 1, and chronic renal failure in Case 2. When prolactin levels were normalized (by withdrawal of risperidone in Case 1 and with cabergoline in Case 2), breast tissue uptake was no longer evident on WBS. CONCLUSION: These cases provide the first documented correlation between serum levels of endogenous prolactin and radioiodine uptake by involuted breast tissue in humans. PMID- 17465869 TI - Lung transplantation for pulmonary metastases and radiation-induced pulmonary fibrosis after radioactive iodine ablation of extensive lung metastases from papillary thyroid carcinoma. PMID- 17465870 TI - Myxedema coma: a well-known but unfamiliar medical emergency. PMID- 17465871 TI - Marine-Lenhart syndrome and radioiodine-131 treatment. PMID- 17465872 TI - Huge cavernous hemangioma of the thyroid gland. PMID- 17465879 TI - Cell type-specific upregulation of Parkin in response to ER stress. AB - Parkin is the gene responsible for a familial form of Parkinson's disease (PD) termed autosomal recessive juvenile parkinsonism (AR-JP)/PARK2. Parkin has been shown to protect cells from endoplasmic reticulum (ER) stress and oxidative stress, presumably due to its ubiquitin ligase (E3) activity that targets proteins for proteasomal degradation. Although the authors showed that parkin is upregulated in response to ER stress, subsequent reports suggest that it does not represent a universal unfolded protein response (UPR). Here the authors report different regulation of parkin in response to ER stress in different cell lines, demonstrating upregulation of parkin as a cell type-specific response to ER stress. 2-Mercaptoethanol (2-ME) and tunicamycin increased the expression of parkin in SH-SY5Y (H) cells, Neuro2a cells, Goto-P3 cells, but not in SH-SY5Y (J) cells and IMR32 cells. In parallel with these studies, similar upregulation of the parkin coregulated gene (PACRG)/gene adjacent to parkin (Glup) was also observed by ER stress. Luciferase assays failed to detect the transcriptional activation of 500 bp parkin/Glup promoter in response to ER stress. These results indicate that induction of parkin by ER stress represents a cell type-specific response. PMID- 17465880 TI - Expanding insights on the involvement of endoplasmic reticulum stress in Parkinson's disease. AB - Parkinson's disease (PD) is the second most common neurodegenerative disease characterized by selective loss of dopaminergic neurons and the presence of Lewy bodies. The pathogenesis of PD remains incompletely understood. Environmental factors, oxidative damage, misfolded protein aggregates, ubiquitin-proteasome system impairment, and mitochondrial dysfunction might all be involved. Recent studies point to activation of endoplasmic reticulum (ER) stress-mediated cell death linked to PD. Accumulation of unfolded and/or misfolded proteins in the ER lumen induces ER stress. To withstand such potentially lethal conditions, intracellular signaling pathways collectively termed the unfolded protein responses (UPR) are activated. The UPR include translational attenuation, induction of ER resident chaperones, and degradation of misfolded proteins through the ER-associated degradation. In case of severe and/or prolonged ER stress, cellular signals leading to cell death are activated. Accumulating evidence suggests that ER stress induced by aberrant protein degradation is implicated in PD. Here the authors review the emerging role of ER stress in PD and related disorders, and highlight current knowledge in this field that may reveal novel insight into disease mechanisms and help to provide novel avenues to potential therapies. PMID- 17465881 TI - Ischemia-induced neuronal cell death and stress response. AB - Neuronal cell death is a major feature of various diseases, including brain ischemia, neuronal degenerative diseases, and traumatic injury, suggesting the importance of investigating the mechanisms that mediate neuronal cell death. Although the various factors that contribute to brain ischemia have been defined and the mechanism through which each factor causes neuronal cell death has been investigated, definite strategies have not been established. In this brief review, we focus on two important mechanisms that contribute to the pathogenesis of brain ischemia. First, we discuss the glutamate theory, a proposed mechanism for the understanding of ischemia-induced neuronal cell death. Second, an accumulation of recent molecular neurobiology evidence regarding the dysfunction of a cellular organelle, the endoplasmic reticulum (ER), suggests that it plays a major role in the pathogenesis of neuronal cell death. Whereas the former theory reflects the role of neuron-specific factors in the induction of cell death, the stress response of the ER for maintenance of its function is regarded as a defense mechanism. Because hypoxia, another major factor in ischemia, results in further dysfunction of the ER, studies on the malfunction of this cellular organelle may facilitate the development of novel strategies to block ischemia induced cell death. PMID- 17465882 TI - Accumulation of misfolded protein through nitrosative stress linked to neurodegenerative disorders. AB - Protein quality control is a critical feature of intracellular homeostasis. In particular, unfolded or misfolded proteins resulting from environmental stresses or free radicals are rapidly degraded via the ubiquitin-proteasome pathway. Nitric oxide (NO), a free radical gas, has been reported to be involved in such processes as vasorelaxation and neurotransmission. Conversely, NO also is implicated in neuronal cell death or neurodegeneration. Recent reports suggest that S-nitrosylation of proteins is a significant cause of neural dysfunction leading to neurodegenerative disorders. Specifically, S-nitrosylation of parkin eventually leads to the accumulation of unfolded proteins and subsequent neuronal death. The focus of this review is the identity of the target of NO. Nitrosative stress prevents normal functioning of the endoplasmic reticulum (ER) via S nitrosylation of protein-disulfide isomerase (PDI), which is located in the ER lumen. This may contribute to the accumulation of misfolded proteins, as well as sustained activation of the unfolded protein response (UPR) pathway. These phenomena may be linked to the development of sporadic neurodegenerative diseases. PMID- 17465883 TI - Does thioredoxin-1 prevent mitochondria- and endoplasmic reticulum-mediated neurotoxicity of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine? AB - We show that 1-methyl-4-phenylpyridinium ion (MPP(+)), an active metabolite of 1 methyl-4-phenyl-1,2,3,6- tetrahydropyridine (MPTP), induces cytotoxicity via endoplasmic reticulum (ER)- and mitochondria-mediated pathways, and thioredoxin-1 (TRX-1), a redox-active protein, prevents MPTP-induced neurotoxicity. TRX-1 overexpression suppressed reactive oxygen species and the ATP decline caused by MPP(+) in HepG2 cells. MPP(+) activated caspase-12 in PC12 cells and induced cytotoxicity in HeLa-rho(0) cells lacking mitochondrial DNA, as well as in the parental HeLa-S3 cells. TRX-1-transgenic mice demonstrated significant resistance to caspase-12 activation and the apoptotic decrease of dopaminergic neurons after MPTP administration, compared with wild-type C57BL/6 mice. PMID- 17465884 TI - Non-thiol reagents regulate ryanodine receptor function by redox interactions that modify reactive thiols. AB - The Ca(2+) release channel (CRC) from sarcoplasmic reticulum (SR) is rich in thiol groups, and their oxidation/- reduction by thiol reagents activates/inhibits the CRC. Most channel regulators are not thiol reagents, and the mechanism of their action is illusive. Here the authors show that many channel activators act as electron acceptors, while many channel inhibitors act as electron donors in free radical reactions. The channel activator, caffeine, and the CRC inhibitor, tetracaine, are shown to interact competitively, which suggests that there exists a common site(s) on the CRC, that integrates the donor/acceptor effects of ligands. Moreover, channel activators shift the redox potential of reactive thiols on the ryanodine receptor (RyR) to more negative values and decrease the number of reactive thiols, while channel inhibitors shift the redox potential to more positive values and increase the number of reactive thiols. These observations suggest that the non-thiol channel modulators shift the thiol-disulfide balance within CRC by transiently exchanging electrons with the Ca(2+) release protein. PMID- 17465885 TI - miRNA: licensed to kill the messenger. AB - Current developments have brought non-coding genes under limelight together with their better-known siblings, the coding genes or mRNA. The 2006 Nobel Prize in Physiology or Medicine was awarded to Andrew Fire and Craig Mello for their 1998 discovery that double-stranded RNA triggers suppression of gene activity in a homology-dependent manner, a process named RNA interference (RNAi). Post transcriptional regulation of genes was generally regarded as an odd regulatory mechanism for several years until it was learnt that regulatory trans-acting antisense RNAs exist in several species. Identification of a large number of small RNA molecules called microRNAs (miRNAs) elevated the overall field of biomedical RNAi to the striking level of current recognition. miRNAs represent a class of endogenous small ( approximately 22 nucleotides) RNA molecules that can repress protein synthesis. It is estimated that there are over 600 miRNAs in mammalian cells, and that about 30% of all genes are regulated by miRNA. Current understanding of the molecular mechanism of any disease would be incomplete without factoring in the functional significance of miRNA. In the category of the futuristic RNAi drugs, miRNA-based therapies are promising. The field has progressed rapidly as it relates to cancer research (highlighted in DNA and Cell Biology Volume 26, Number 4), while development in most other areas (highlighted in DNA and Cell Biology Volume 26, Number 3) of biomedical research remains in its infancy, offering significant opportunity for researchers. Approaches to interfere with miRNA function in vivo offer novel therapeutic opportunities. Lessons in gene therapy have taught us that tinkering with the genetic machinery comes with its own set of risks, especially in a clinical setting. miRNA-based therapies are also subject to such risks, which need to be prudently managed. Having acknowledged the potential risk, we have to recognize that new knowledge about the functional roles of miRNA is revolutionizing cell biology and will have a major impact on biomedical research imminently. PMID- 17465886 TI - Intronic microRNA: discovery and biological implications. AB - Identification of microRNAs (miRNAs) is essential to studying their physiological functions. Due to the difficulties in discovering truly expressed miRNAs from genomic random hairpin secondary structure sequences, it is beneficial to predict them from expressed sequences--expressed sequence tags (ESTs) and intronic sequences. We used a modified scanning pipeline using criteria based on the features of known pre-miRNAs and phylogenetic conservation for predicting intronic miRNAs. Upon examination, we found that 25% of known human miRNAs belong to intronic regions of known protein-coding genes. About 50% of these intronic miRNAs reside in introns whose length is longer than 5,000 bps. It is likely that these intronic miRNAs can have their own independently regulated transcription units, which can be regulated by RNA polymerase II (Pol II) or RNA polymerase III (Pol III). It was recently demonstrated that RNA Pol III could transcribe human miRNAs through associated repetitive elements. Since various repetitive elements are often found to be present in the intronic regions, the distribution of intronic miRNAs and their possible transcription regulation are presented. Although the intronic miRNAs and their host genes could be regulated independently, it is possible that the intronic miRNA can still down-regulate its own host protein-coding gene by targeting the untranslated region (UTR) of the host gene. Another biological implication is that intronic miRNAs could play an important role as negative feedback regulators. We propose hypothetical models of such feedback regulation on host protein-coding genes by selecting the transcription factors as miRNA targets or by protein-protein interactions between intronic miRNA host gene product and miRNA target gene products. PMID- 17465887 TI - Evolutionary conservation of microRNA regulatory circuits: an examination of microRNA gene complexity and conserved microRNA-target interactions through metazoan phylogeny. AB - During the last decade, a variety of critical biological processes, including early embryo development, cell proliferation, differentiation, apoptosis, and metabolic regularity, have been shown to be genetically regulated by a large gene family encoding a class of tiny RNA molecules termed microRNAs (miRNAs). All miRNAs share a common biosynthetic pathway and reaction mechanisms. The sequence of many miRNAs is found to be conserved, in their mature form, among different organisms. In addition, the evolutionary appearance of multicellular organisms appears to correlate with the appearance of the miRNA pathway for regulating gene expression. The miRNA pathway has the potential to regulate vast networks of gene products in a coordinate manner. Recent evidence has not only implicated the miRNA pathway in regulating a vast array of basic cellular processes but also specialized processes that are required for cellular identity and tissue specificity. A survey of the literature shows that some miRNA pathways are conserved virtually intact throughout phylogeny while miRNA diversity also correlates with speciation. The number of miRNA genes, the expression of miRNAs, and target diversities of miRNAs tend to be positively correlated with morphological complexities observed in animals. Thus, organismal complexity can be estimated by the complexity of the miRNA circuitry. The complexity of the miRNA gene families establishes a link between genotypic complexity and phenotypic complexity in animal evolution. In this paper, we start with the discussion of miRNA conservation. Then we interpret the trends in miRNA conservation to deduce miRNA evolutionary trends in metazoans. Based on these conservation patterns observed in each component of the miRNA regulatory system, we attempt to propose a global insight on the probable consistency between morphological evolution in animals and the molecular evolution of miRNA gene activity in the cell. PMID- 17465888 TI - MicroRNAs in skeletal and cardiac muscle development. AB - MicroRNAs (miRNAs) are a recently discovered class of small non-coding RNAs, which are approximately 22 nucleotides in length. miRNAs negatively regulate gene expression by translational repression and target mRNA degradation. It has become clear that miRNAs are involved in many biological processes, including development, differentiation, proliferation, and apoptosis. Interestingly, many miRNAs are expressed in a tissue-specific manner and several miRNAs are specifically expressed in cardiac and skeletal muscles. In this review, we focus on those miRNAs that have been shown to be involved in muscle development. Compelling evidences have demonstrated that muscle miRNAs play an important role in the regulation of muscle proliferation and differentiation processes. However, it appears that miRNAs are not essential for early myogenesis and muscle specification. Importantly, dysregulation of miRNAs has been linked to muscle related diseases, such as cardiac hypertrophy. A mutation resulting in a gain-of function miRNA target site in the myostatin gene leads to down regulation of the targeted protein in Texel sheep. miRNAs therefore are a new class of regulators of muscle biology and they might become novel therapeutic targets in muscle related human diseases. PMID- 17465889 TI - MicroRNA in cutaneous wound healing: a new paradigm. AB - Repair of a defect in the human skin is a highly orchestrated physiological process involving numerous factors that act in a temporally resolved synergistic manner to re-establish barrier function by regenerating new skin. The inducible expression and repression of genes represents a key component of this regenerative process. MicroRNAs (miRNAs) are approximately 22-nucleotide-long endogenously expressed non-coding RNAs that regulate the expression of gene products by inhibition of translation and/or transcription in animals. miRNAs play a key role in skin morphogenesis and in regulating angiogenesis. The vascular endothelial growth factor signaling path seems to be under repressor control by miRNAs. Mature miRNA-dependent mechanisms impair angiogenesis in vivo. It is critically important to recognize that the understanding of cutaneous wound healing is incomplete without appreciating the functional significance of wound induced miRNA. Ongoing work in our laboratory has led to the observation that the cutaneous wound healing process involves changes in the expression of specific miRNA at specific phases of wound healing. We hypothesize that dysregulation of specific miRNA is critical in derailing the healing sequence in chronic problem wounds. If tested positive, this hypothesis is likely to lead to completely novel diagnostic and therapeutic strategies for the treatment of problem wounds. PMID- 17465890 TI - MicroRNAs in viral replication and pathogenesis. AB - MicroRNAs (miRNAs) are an important class of small, noncoding, regulatory RNAs found to be involved in regulating a wide variety of important cellular processes by the sequence-specific inhibition of gene expression. Viruses have evolved a number of mechanisms to take advantage of the regulatory potential of this highly conserved, ubiquitous pathway known as RNA interference (RNAi). This review will focus on the recent efforts to understand the complex relationship between vertebrate viruses and the RNAi pathway, as well as the role of silencing pathways in the inhibition of pathogenic genetic elements, including transposons and retrotransposons. PMID- 17465891 TI - In situ detection of animal and plant microRNAs. AB - MicroRNAs (miRNAs) are recently discovered short regulatory RNA molecules representing a new layer in posttranscriptional gene expression regulation. Although more than 450 human miRNAs have been identified, only a very few of them have been characterized in detail. The precise understanding of miRNA-mediated processes requires the reliable spatial and temporal analyses of miRNA accumulation at tissue/cell level. However, the detection of miRNAs by in situ hybridization (ISH) is technically challenging because of the small size of target sequences. It was shown recently that locked nucleic acid nucleotide containing probes can anneal to short nucleic acids with high specificity. This enabled several research groups to analyze the expression patterns of miRNAs in both plant and animal tissues. This review focuses on the results of recent publications on the detection of miRNAs by ISH. PMID- 17465892 TI - MicroRNAs and regenerative medicine. AB - MicroRNAs, identified only relatively recently, are regulators of gene expression with potential medical benefits. The combination of microRNAs and regenerative medicine is an emerging interdisciplinary medical field that can yield exciting new possibilities for clinical medicine. In this paper, we review the prospects of microRNAs as future therapies in regenerative medicine. Recently, researchers have demonstrated the crucial roles of microRNAs, not only in the differentiation and proliferation of stem cells, which have a key function in the regeneration and transplantation of organs, but also in oncogenesis. Several lines of indirect evidence show that the initiation and maintenance of cancer stem cells might also be under the control of microRNAs. Further, microRNAs have been indicated to be involved in diverse biological processes, suggesting the potential role of these molecules in the treatment of diseases. PMID- 17465893 TI - Algorithms for mapping of mRNA targets for microRNA. AB - MicroRNAs (miRNAs) are involved in human health and disease as endogenous suppressors of the translation of coding genes. At this early point of time in miRNA biology, it is important to identify specific cognate mRNA targets for miRNA. Investigation of the significance of miRNAs in disease processes relies on algorithms that hypothetically link specific miRNAs to their putative target genes. The development of such algorithms represents a hot area of research in biomedical informatics. Lack of biological data linking specific miRNAs to their respective mRNA targets represents the most serious limitation at this time. This article presents a concise review addressing the most popular concepts underlying state-of-the-art algorithms and principles aimed at target mapping for specific miRNAs. Strategies for improvement of the current bioinformatics tools and effective approaches for biological validation are discussed. PMID- 17465894 TI - The evolutionary ecology of individual phenotypic plasticity in wild populations. AB - The ability of individual organisms to alter morphological and life-history traits in response to the conditions they experience is an example of phenotypic plasticity which is fundamental to any population's ability to deal with short term environmental change. We currently know little about the prevalence, and evolutionary and ecological causes and consequences of variation in life history plasticity in the wild. Here we outline an analytical framework, utilizing the reaction norm concept and random regression statistical models, to assess the between-individual variation in life history plasticity that may underlie population level responses to the environment at both phenotypic and genetic levels. We discuss applications of this framework to date in wild vertebrate populations, and illustrate how natural selection and ecological constraint may alter a population's response to the environment through their effects at the individual level. Finally, we present future directions and challenges for research into individual plasticity. PMID- 17465895 TI - Cost of resistance to parasites in digital organisms. AB - Virtually all organisms are attacked by parasites and are therefore expected to evolve resistance against these natural enemies. Parasite resistance is costly in a wide range of organisms, although the generality of such costs has been questioned, especially when resistance is not based on reallocation of resources. Digital organisms are increasingly used to explore aspects of life in general. In the Tierra system, there is a trade-off between resistance against parasites and competitive ability. Because digital organisms are too simple to store resources, the finding that resistance to parasites is costly in digital organisms suggests that costs of parasite resistance can also occur when resistance is not resource based. PMID- 17465896 TI - Direct benefits and costs for hybridizing Ficedula flycatchers. AB - It is well understood that females may gain direct benefits from breeding with attractive males. However, the direct fitness effects of mate-choice are rarely considered with respect to mating between different species (hybridization), a field dominated by discussion of indirect costs of producing unfit hybrid offspring. Hybridizing females may also gain by the types of direct benefits that are important for intraspecific mate choice, and in addition may have access to certain benefits that are restricted to mating with males of an ecologically diverged sister-taxon. We investigate possible direct benefits and costs female Ficedula flycatchers gain from breeding with a heterospecific male, and demonstrate that hybridizing female collared flycatchers (F. albicollis) breed in territories that do not suffer the seasonal decline in habitat quality experienced by females breeding with conspecifics. We exclude the hypotheses that heterospecific males provide alternative food-types or assume a greater amount of the parental workload. In fact, the diets of the two species (F. albicollis and F. hypoleuca) were highly similar, suggesting possible interspecific competition over food resources in sympatry. We discuss the implications of direct fitness effects of hybridization, and why there has been such a disparity in the attention paid to such benefits and costs with regard to intraspecific and interspecific mate-choice. PMID- 17465897 TI - No genetic evidence of sex-biased dispersal in a lekking bird, the capercaillie (Tetrao urogallus). AB - Sex-biased dispersal is often connected to the mating behaviour of the species. Even if patterns of natal dispersal are reasonably well documented for monogamous birds, only a few data are available for polygynous and especially lekking species. We investigated the dispersal of the capercaillie (Tetrao urogallus) by examining sex-specific gene flow among the leks. Genetic information was extracted using nuclear and mitochondrial molecular markers for sexed faecal samples and analysed by novel Bayesian statistical methods. Contrary to the traditional view that the males are highly philopatric and female is the dispersing sex, we found roughly equivalent gross and effective dispersal of the sexes. The level of polygamy has a strong influence on the effective population size and on the effective dispersal. The results do not support the theories that dispersal evolves solely as a result of resource competition or other advantages to males obtained through kin selection in lekking species. PMID- 17465898 TI - Food availability affects the maternal transfer of androgens and antibodies into eggs of a colonial seabird. AB - Mothers can improve the quality of their offspring by increasing the level of certain components in their eggs. To examine whether or not mothers increase deposition of such components in eggs as a function of food availability, we food supplemented black-legged kittiwake females (Rissa tridactyla) before and during egg laying and compared deposition of androgens and antibodies into eggs of first and experimentally induced replacement clutches. Food-supplemented females transferred lower amounts of androgens and antibodies into eggs of induced replacement clutches than did non-food-supplemented mothers, whereas first clutches presented no differences between treatments. Our results suggest that when females are in lower condition, they transfer more androgens and antibodies into eggs to facilitate chick development despite potential long-term costs for juveniles. Females in prime condition may avoid these potential long-term costs because they can provide their chicks with more and higher quality resources. PMID- 17465899 TI - Genotypic selection in Daphnia populations consisting of inbred sibships. AB - The genetic basis of fitness reduction associated with inbreeding is still poorly understood. Here we use associations between allozyme genotypes and fitness to investigate the genetic basis of inbreeding depression in experimental outdoor populations of the water flea, Daphnia magna. In Daphnia, a phase of clonal reproduction follows hatching from sexually produced resting eggs, and changes in genotype frequencies during the clonal phase can be used to estimate fitness. Our experiment resembles natural colonization of ponds in that single clones colonize an empty pool, expand asexually and produce sexual offspring by selfing (sisters mate with their clonal brothers). These offspring diapause and form populations consisting of selfed sibships in the following spring. In 12 of 13 experimental populations, genotypes of selfed hatchlings after diapause conformed to Mendelian expectations. During the subsequent ca. 10 asexual generations, however, genotype frequencies changed significantly at 19 of 27 single loci studied within populations, mostly in favour of heterozygotes, with heterozygosity at multiple loci affecting the change in genotype frequency multiplicatively. Because variance in heterozygosity among siblings at a given marker reflects only heterozygosity in the chromosomal region around this marker, our results suggest that selection at fitness-associated loci in the chromosomal regions near the markers were responsible for these changes. The genotype frequency changes were more consistent with selection acting on linked loci than on the allozymes themselves. Taken together, the evidence for abundant selection in the chromosomal regions of the markers and the fact that changes in genotype frequencies became apparent only after several generations of clonal selection, point to a genetic load consisting of many alleles of small or intermediate effects, which is consistent with the strong genetic differentiation and repeated genetic bottlenecks in the metapopulation from which the animals for this study were obtained. PMID- 17465900 TI - Rapid population divergence in thermal reaction norms for an invading species: breaking the temperature-size rule. AB - The temperature-size rule is a common pattern of phenotypic plasticity in which higher temperature during development results in a smaller adult body size (i.e. a thermal reaction norm with negative slope). Examples and exceptions to the rule are known in multiple groups of organisms, but rapid population differentiation in the temperature-size rule has not been explored. Here we examine the genetic and parental contributions to population differentiation in thermal reaction norms for size, development time and survival in the Cabbage White Butterfly Pieris rapae, for two geographical populations that have likely diverged within the past 150 years. We used split-sibship experiments with two temperature treatments (warm and cool) for P. rapae from Chapel Hill, NC, and from Seattle, WA. Mixed-effect model analyses demonstrate significant genetic differences between NC and WA populations for adult size and for thermal reaction norms for size. Mean adult mass was 12-24% greater in NC than in WA populations for both temperature treatments; mean size was unaffected or decreased with temperature (the temperature-size rule) for the WA population, but size increased with temperature for the NC population. Our study shows that the temperature-size rule and related thermal reaction norms can evolve rapidly within species in natural field conditions. Rapid evolutionary divergence argues against the existence of a simple, general mechanistic constraint as the underlying cause of the temperature size rule. PMID- 17465901 TI - Predominance of outcrossing in Lymnaea stagnalis despite low apparent fitness costs of self-fertilization. AB - We have quantified the natural mating system in eight populations of the simultaneously hermaphroditic aquatic snail Lymnaea stagnalis, and studied the ecological and genetic forces that may be directing mating system evolution in this species. We investigated whether the natural mating system can be explained by the availability of mates, by the differential survival of self- and cross fertilized snails in nature, and by the effects of mating system on parental fecundity and early survival. The natural mating system of L. stagnalis was found to be predominantly cross-fertilizing. Density of snails in the populations had no relationship with the mating system, suggesting that outcrossing rates are not limited by mate availability at the population densities observed. Contrary to expectations for outcrossing species, we detected no evidence for inbreeding depression in survival in nature with inferential population genetic methods. Further, experimental manipulations of mating system in the laboratory revealed that self-fertilization had no effect on parental fecundity, and only minor effects on offspring survival. Predominance of cross-fertilization despite low apparent fitness costs of self-fertilization is at odds with the paradigm that high self-fertilization depression is necessary for maintenance of cross fertilization in self-compatible hermaphrodites. PMID- 17465902 TI - Invasion and fixation of sex-reversal genes. AB - We simulated a meta-population with random dispersal among demes but local mating within demes to investigate conditions under which a dominant female-determining gene W, with no individual selection advantage, can invade and become fixed in females, changing the population from male to female heterogamety. Starting with one mutant W in a single deme, the interaction of sex ratio selection and random genetic drift causes W to be fixed among females more often than a comparable neutral mutation with no influence on sex determination, even when YY males have slightly reduced viability. Meta-population structure and interdeme selection can also favour the fixation of W. The reverse transition from female to male heterogamety can also occur with higher probability than for a comparable neutral mutation. These results help to explain the involvement of sex-determining genes in the evolution of sex chromosomes and in sexual selection and speciation. PMID- 17465903 TI - Inference of parasite local adaptation using two different fitness components. AB - Estimating parasite fitness is central to studies aiming to understand parasite evolution. Theoretical models generally use the basic reproductive rate R(0) to express fitness, yet it is very difficult to quantify R(0) empirically and experimental studies often use fitness components such as infection intensity or infectivity as substitutes. These surrogate measures may be biased in several ways. We assessed local adaptation of the microsporidium Ordospora colligata to its host, the crustacean Daphnia magna using two different parasite fitness components: infection persistence over several host generations in experimental populations and infection intensity in individual hosts. We argue that infection persistence is a close estimator of R(0), whereas infection intensity measures only a component of it. Both measures show a pattern that is consistent with parasite local adaptation and they correlate positively. However, several inconsistencies between them suggest that infection intensity may at times provide an inadequate estimate of parasite fitness. PMID- 17465904 TI - Revisiting Jablonski (1993): cladogenesis and range expansion explain latitudinal variation in taxonomic richness. AB - The increase in diversity towards the equator arises from latitudinal variation in rates of cladogenesis, extinction, immigration and/or emigration of taxa. We tested the relative contribution of all four processes to the latitudinal gradient in 26 marine invertebrate orders with extensive fossil records, examined previously by David Jablonski. Coupling Jablonski's estimates of latitudinal variation in cladogenesis with new data on patterns of extinction and current distributions, we show that the present-day gradient in diversity is caused by higher rates of cladogenesis and subsequent range expansion (immigration) at lower latitudes. In contrast, extinction and emigration were not important in the creation of the latitudinal gradient in ordinal richness. This work represents one of the first simultaneous tests of the role of all four processes in the creation of the latitudinal gradient in taxonomic richness, and suggests that low tropical extinction rates are not essential to the creation of latitudinal diversity gradients. PMID- 17465905 TI - The evolution of conspecific gamete precedence and its effect on reinforcement. AB - Conspecific gamete precedence, the usage of conspecific sperm by a female that mates with both a conspecific and a heterospecific male, has been found in many taxa. We construct a population genetic model to examine the evolution of conspecific gamete precedence and its coevolution with premating isolation in the process of reinforcement. Our findings suggest that conspecific gamete precedence can evolve via a process very similar to reinforcement. We explore the nature of the selection against hybridization necessary to drive this evolution. Moreover, our results confirm the prediction of Marshall et al. (Trends Ecol. Evol. 2002;17:558-563) that conspecific gamete precedence will inhibit the evolution of reinforcement between two species. We further find that reinforcement will inhibit the evolution of conspecific gamete precedence. Both reinforcement and conspecific gamete precedence increase reproductive isolation and contribute to the process of speciation. We discuss factors that may affect which of these phenomena are likely to become predominant between incipient species. PMID- 17465906 TI - Interspecific variation in egg testosterone levels: implications for the evolution of bird song. AB - Although interspecific variation in maternal effects via testosterone levels can be mediated by natural selection, little is known about the evolutionary consequences of egg testosterone for sexual selection. However, two nonexclusive evolutionary hypotheses predict an interspecific relationship between egg testosterone levels and the elaboration of sexual traits. First, maternal investment may be particularly enhanced in sexually selected species, which should generate a positive relationship. Secondly, high prenatal testosterone levels may constrain the development of sexual characters, which should result in a negative relationship. Here we investigated these hypotheses by exploring the relationship between yolk testosterone levels and features of song in a phylogenetic study of 36 passerine species. We found that song duration and syllable repertoire size were significantly negatively related to testosterone levels in the egg, even if potentially confounding factors were held constant. These relationships imply that high testosterone levels during early development of songs may be detrimental, thus supporting the developmental constraints hypothesis. By contrast, we found significant evidence that song-post exposure relative to the height of the vegetation is positively related to egg testosterone levels. These results support the hypothesis that high levels of maternal testosterone have evolved in species with intense sexual selection acting on the location of song-posts. We found nonsignificant effects for intersong interval and song type repertoire size, which may suggest that none of the above hypothesis apply to these traits, or they act simultaneously and have opposing effects. PMID- 17465907 TI - A screen for immunity genes evolving under positive selection in Drosophila. AB - Genes involved in the immune system tend to have higher rates of adaptive evolution than other genes in the genome, probably because they are coevolving with pathogens. We have screened a sample of Drosophila genes to identify those evolving under positive selection. First, we identified rapidly evolving immunity genes by comparing 140 loci in Drosophila erecta and D. yakuba. Secondly, we resequenced 23 of the fastest evolving genes from the independent species pair D. melanogaster and D. simulans, and identified those under positive selection using a McDonald-Kreitman test. There was strong evidence of adaptive evolution in two serine proteases (persephone and spirit) and a homolog of the Anopheles serpin SRPN6, and weaker evidence in another serine protease and the death domain protein dFADD. These results add to mounting evidence that immune signalling pathway molecules often evolve rapidly, possibly because they are sites of host parasite coevolution. PMID- 17465908 TI - Parallel evolution of lake whitefish dwarf ecotypes in association with limnological features of their adaptive landscape. AB - Sympatric fish populations observed in many north temperate lakes are among the best models to study the processes of population divergence and adaptive radiation. Despite considerable research on such systems, little is known about the associations between ecological conditions and the extent of ecotypic divergence. In this study, we examined the biotic and abiotic properties of postglacial lakes in which lake whitefish, Coregonus clupeaformis, occur as a derived dwarf ecotype in sympatry with an ancestral normal ecotype. We compared 19 limnological variables between two groups of lakes known from previous studies to harbour sympatric dwarf and normal ecotypes with high and low levels of phenotypic and genetic differentiation respectively. We found clear environmental differences between the two lake groups. Namely, oxygen was the most discriminant variable, where lakes harbouring the most divergent populations were characterized by the greatest hypolimnetic oxygen depletion. These lakes also had lower zooplankton densities and a narrower distribution of zooplantonic prey length. These results suggest that the highest differentiation between sympatric ecotypes occurs in lakes with reduced habitat and prey availability that could increase competition for resources. This in turns supports the hypothesis that parallelism in the extent of phenotypic divergence among sympatric whitefish ecotypes is associated with parallelism in adaptive landscape in terms of differences in limnological characteristics, as well as availability and structure of the zooplanktonic community. PMID- 17465909 TI - The genetic architecture of a niche: variation and covariation in host use traits in the Colorado potato beetle. AB - The genetic basis of host plant use by phytophagous insects can provide insight into the evolution of ecological niches, especially phenomena such as specialization and phylogenetic conservatism. We carried out a quantitative genetic analysis of multiple host use traits, estimated on five species of host plants, in the Colorado potato beetle, Leptinotarsa decemlineata (Coleoptera: Chrysomelidae). Mean values of all characters varied among host plants, providing evidence that adaptation to plants may require evolution of both behavioral (preference) and post-ingestive physiological (performance) characteristics. Significant additive genetic variation was detected for several characters on several hosts, but not in the capacity to use the two major hosts, a pattern that might be caused by directional selection. No negative genetic correlations across hosts were detected for any 'performance' traits, i.e. we found no evidence of trade-offs in fitness on different plants. Larval consumption was positively genetically correlated across host plants, suggesting that diet generalization might evolve as a distinct trait, rather than by independent evolution of feeding responses to each plant species, but several other traits did not show this pattern. We explored genetic correlations among traits expressed on a given plant species, in a first effort to shed light on the number of independent traits that may evolve in response to selection for host-plant utilization. Most traits were not correlated with each other, implying that adaptation to a novel potential host could be a complex, multidimensional 'character' that might constrain adaptation and contribute to the pronounced ecological specialization and the phylogenetic niche conservatism that characterize many clades of phytophagous insects. PMID- 17465910 TI - Pheromonal dominance and the selection of a socially parasitic honeybee worker lineage (Apis mellifera capensis Esch.). AB - The recent invasion by self-replicating socially parasitic Cape honeybee workers, Apis mellifera capensis, of colonies of the neighbouring African subspecies Apis mellifera scutellata represents an opportunity to study evolution of intraspecific parasitism in real time. As honeybee workers compete pheromonally for reproductive dominance, and as A. m. capensis workers readily produce queen like pheromones, we hypothesized that these semiochemicals promoted the evolution of intraspecific social parasitism. Remarkably, the offspring of a single worker became established as a parasite in A. m. scutellata's range. This could have resulted from extreme selection among different clonal parasitic worker lineages. Using pheromonal contest experiments, we show that the selected parasitic lineage dominates in the production of mandibular gland pheromones over all other competitors to which it is exposed. Our results suggest that mandibular gland pheromones played a key role in the evolution of intraspecific social parasitism in the honeybee and in the selection of a single genotype of parasitic workers. PMID- 17465911 TI - The evolution of sperm morphometry in pheasants. AB - Post-copulatory sexual selection is thought to be a potent evolutionary force driving the diversification of sperm shape and function across species. In birds, insemination and fertilization are separated in time and sperm storage increases the duration of sperm-female interaction and hence the opportunity for sperm competition and cryptic female choice. We performed a comparative study of 24 pheasant species (Phasianidae, Galliformes) to establish the relative importance of sperm competition and the duration of sperm storage for the evolution of sperm morphometry (i.e. size of different sperm traits). We found that sperm size traits were negatively associated with the duration of sperm storage but were independent of the risk of sperm competition estimated from relative testis mass. Our study emphasizes the importance of female reproductive biology for the evolution of sperm morphometry particularly in sperm-storing taxa. PMID- 17465912 TI - Mechanism of a plastic phenotypic response: predator-induced shell thickening in the intertidal gastropod Littorina obtusata. AB - Phenotypic plasticity has been the object of considerable interest over the past several decades, but in few cases are mechanisms underlying plastic responses well understood. For example, it is unclear whether predator-induced changes in gastropod shell morphology represent an active physiological response or a by product of reduced feeding. We address this question by manipulating feeding and growth of intertidal snails, Littorina obtusata, using two approaches: (i) exposure to predation cues from green crabs Carcinus maenas and (ii) reduced food availability, and quantifying growth in shell length, shell mass, and body mass, as well as production of faecal material and shell micro-structural characteristics (mineralogy and organic fraction) after 96 days. We demonstrate that L. obtusata actively increases calcification rate in response to predation threat, and that this response entails energetic and developmental costs. That this induced response is not strictly tied to the animal's behaviour should enhance its evolutionary potential. PMID- 17465913 TI - Increase in song frequency decreases spermatophore size: correlative evidence of a macroevolutionary trade-off in katydids (Orthoptera: Tettigoniidae). AB - In many katydids, the male feeds his mate with a large gelatinous spermatophore. Males of most species also produce elaborate calling songs. We predicted a negative relationship between spermatophore size and call frequency because of trade-offs between these two costly traits. Our comparative analysis controlling phylogeny and body size supported this prediction. Although call frequency is expected to decrease with increasing body size, after controlling for phylogeny, both variables were not related. Finally, given that song frequency and spermatophore size are likely targets of sexual selection, we examined the relationship between these variables and sexual size dimorphism (SSD) which can be influenced by sexual selection on body size. We found that only female body size was positively related to SSD, suggesting that natural and/or sexual selection on female body size may be stronger than sexual selection on male and spermatophore size. PMID- 17465914 TI - Evolution of viviparity in warm-climate lizards: an experimental test of the maternal manipulation hypothesis. AB - The maternal manipulation hypothesis for the evolution of reptilian viviparity has been claimed to apply to any situation where gravid females are able to maintain body temperatures different from those available in external nests, but empirical data that support this hypothesis are very limited. Here, we tested this hypothesis using gravid females of a warm-climate lizard, Mabuya multifasciata, by subjecting them to five thermal regimes for the whole gestation period. We found gravid females selected lower body temperatures and thermoregulated more precisely than did nongravid females. Offspring produced in different treatments differed in head size, limb length and sprint speed, but not in overall body size or mass. Variation in morphological traits of offspring was induced primarily by extreme temperatures. Sprint speed of offspring was more likely affected by the mean but not by the variance of gestation temperatures. Gravid females maintained more stable body temperatures than did nongravid females not because these temperatures resulted in the optimization of offspring phenotypes but because the range of temperatures optimal for embryonic development was relatively narrow. Our data conform to the main predictions from the maternal manipulation hypothesis that females should adjust thermoregulation during pregnancy to provide optimal thermal conditions for developing embryos and that phenotypic traits forged by maternal thermoregulation should enhance offspring fitness. PMID- 17465915 TI - Resource-dependent sex-allocation in a simultaneous hermaphrodite. AB - Most sex allocation theory is based on the relationship between the resource investment into male and female reproduction and the consequent fitness returns (often called fitness-gain curves). Here we investigate the effects of resource availability on the sex allocation of a simultaneously hermaphroditic animal, the free-living flatworm Macrostomum lignano. We kept the worms under different resource levels and determined the size of their testes and ovaries over a period of time. At higher resource levels, worms allocated relatively more into the female function, suggesting a saturating male fitness-gain curve for this species. A large part of the observed effect was due to a correlated increase in body size, showing size-dependent sex allocation in M. lignano. However, a significant part of the overall effect was independent of body size, and therefore likely due to the differences in resource availability. Moreover, in accordance with a saturating male fitness-gain curve, the worms developed the male gonads first. As the group size was kept constant, our results contrast with expectations from sex allocation models that deal with local mate competition alone, and with previous experiments that test these models. PMID- 17465916 TI - The distribution of copia-type retrotransposons and the evolutionary history of tomato and related wild species. AB - Retrotransposons are mobile genetic elements that amplify throughout the genome and may be important contributors of genetic diversity. Their distribution is influenced by element behaviour and host-driven controls. We analysed the distribution of three copia-type retrotransposons, ToRTL1, T135 and Tnt1 using sequence-specific amplification polymorphism in self-compatible (SC) and incompatible (SI) species of Solanum subsection Lycopersicon, and genetically mapped polymorphic insertions in S. lycopersicum (tomato). The majority of polymorphic insertions (61%) are located in centromeric regions of the tomato genome. A significant positive relationship was detected between insertion polymorphisms and mating system, independent of selection as most insertions were found to be neutral. As insertion patterns successfully inferred interspecific relationships of Solanum subsection Lycopersicon, our results suggest that the distribution of ToRTL1, T135 and Tnt1 may essentially be determined by selection removing strongly deleterious insertions, with genetic drift and mating system, but not recombination rate, playing important roles. PMID- 17465917 TI - Sex-specific ecomorphological variation and the evolution of sexual dimorphism in dwarf chameleons (Bradypodion spp.). AB - Natural selection can influence the evolution of sexual dimorphism through selection for sex-specific ecomorphological adaptations. The role of natural selection in the evolution of sexual dimorphism, however, has received much less attention than that of sexual selection. We examined the relationship between habitat structure and both male and female morphology, and sexual dimorphism in size and shape, across 21 populations of dwarf chameleon (genus Bradypodion). Morphological variation in dwarf chameleons was strongly associated with quantitative, multivariate aspects of habitat structure and, in most cases, relationships were congruent between the sexes. However, we also found consistent relationships between habitat and sexual dimorphism. These resulted from both differences in magnitude of ecomorphological relationships that were otherwise congruent between the sexes, as well as in sex-specific ecomorphological adaptations. Our study provides evidence that natural selection plays an important role in the evolution of sexual dimorphism. PMID- 17465918 TI - An unexpected influence of widely used significance thresholds on the distribution of reported P-values. AB - We consider the problematic relationship between publication success and statistical significance in the light of analyses in which we examine the distribution of published probability (P) values across the statistical 'significance' range, below the 5% probability threshold. P-values are often judged according to whether they lie beneath traditionally accepted thresholds (< 0.05, < 0.01, < 0.001, < 0.0001); we examine how these thresholds influence the distribution of reported absolute P-values in published scientific papers, the majority in biological sciences. We collected published P-values from three leading journals, and summarized their distribution using the frequencies falling across and within these four threshold values between 0.05 and 0. These published frequencies were then fitted to three complementary null models which allowed us to predict the expected proportions of P-values in the top and bottom half of each inter-threshold interval (i.e. those lying below, as opposed to above, each P-value threshold). Statistical comparison of these predicted proportions, against those actually observed, provides the first empirical evidence for a remarkable excess of probability values being cited on, or just below, each threshold relative to the smoothed theoretical distributions. The pattern is consistent across thresholds and journals, and for whichever theoretical approach used to generate our expected proportions. We discuss this novel finding and its implications for solving the problems of publication bias and selective reporting in evolutionary biology. PMID- 17465919 TI - Assessing the speed and predictability of local adaptation in invasive California poppies (Eschscholzia californica). AB - Insight into the speed and predictability of local adaptation can be gained by studying organisms, such as invasive species, that have recently expanded their geographical ranges. Common garden studies were designed to address these issues with the California poppy, Eschscholzia californica, collected from a wide range of environments in both its native (California) and invasive (Chile) ranges. We found similar patterns of plant trait variation along similar abiotic gradients in plants collected from both areas. Multivariate analysis demonstrated that coastal plants from both areas tended to be shorter, smaller plants with smaller seeds and flowers that germinate and flower later than plants collected from inland locations. In addition, size and fecundity traits in both native and invasive poppies were correlated with average rainfall totals; the plants that grew the largest and were the most fecund during the first year of growth originated from the driest areas. This parallel variation suggests that these traits are adaptive and that these patterns have evolved in Chile during the 110 150 years since introduction. PMID- 17465920 TI - Morphological evidence of correlational selection and ecological segregation between dextral and sinistral forms in a polymorphic flatfish, Platichthys stellatus. AB - Phenotypic polymorphisms in natural systems are often maintained by ecological selection, but only if niche segregation between morphs exists. Polymorphism for eyed-side direction is rare among the approximately 700 species of flatfish (Pleuronectiformes), and the evolutionary mechanisms that maintain it are unknown. Platichthys stellatus (starry flounder) is a polymorphic pleuronectid flatfish exhibiting large, clinal variation in proportion of left-eyed (sinistral) morphs, from 50% in California to 100% in Japan. Here I examined multiple traits related to swimming and foraging performance between sinistral and dextral morphs of P. stellatus from 12 sites to investigate if the two morphs differ in ways that may affect function and ecology. Direction of body asymmetry was correlated with several other characters: on an average, dextral morphs had longer, wider caudal peduncles, shorter snouts and fewer gill rakers than sinistral morphs. Although the differences were small in magnitude, they were consistent in direction across samples, implying that dextral and sinistral starry flounder may be targeting different prey types. Morphological differences between morphs were greatest in samples where the chances of competitive interactions between them were the greatest. These results suggest that the two morphs are not ecologically identical, may represent a rare example of divergent selection maintaining polymorphism of asymmetric forms, and that correlational selection between body asymmetry and other characters may be driven by competitive interactions between sinistral and dextral flatfish. This study is one of very few that demonstrates the ecological significance of direction in a species with polymorphic asymmetric forms. PMID- 17465921 TI - Polyandry in coal tits Parus ater: fitness consequences of putting eggs into multiple genetic baskets. AB - Females of many species mate with multiple males within a single reproductive cycle. One hypothesis to explain polyandry postulates that females benefit from increasing within-brood genetic diversity. Two mechanisms may render sire genetic diversity beneficial for females, genetic bet-hedging vs. non-bet-hedging. We analysed whether females of the socially monogamous coal tit (Parus ater) benefit via either of these mechanisms when engaging in extra-pair (i.e. polyandrous) mating. To obtain a measure of within-brood genetic diversity as a function of paternal genetic contributions, we calculated a sire diversity index based on the established Shannon-Wiener Index. In 246 broods from two consecutive years, sire genetic diversity had no effect on either the mean or the variance in brood fitness measured as offspring recruitment within 4 years after birth. The hypothesis that benefits of increasing sire diversity contribute to selection for female extra-pair mating behaviour in P. ater was therefore not supported. PMID- 17465922 TI - Body shape vs. colour associated initial divergence in the Telmatherina radiation in Lake Matano, Sulawesi, Indonesia. AB - Highly polymorphic colouration patterns are often associated with sexual selection in fish and can be the initial cause of divergence among closely related taxa. Here we use genetic, body colour and geometric morphometric data collected on 118 fish from Lake Matano, Sulawesi, Indonesia to test if colouration is the initial cause of divergence in the radiating Telmatherina genus. Results reveal that all Telmatherina previously described in this system can be categorized into three mitochondrial lineages and that colouration is only weakly associated with early divergence. Clade-specific body shapes, however, likely adapted to microenvironments are key to the initial divergence in this system. Data also show that although colourations were not likely instrumental in seeding divergence in these fish, they appear to have developed in parallel within each clade. Our results are consistent with an emerging pattern repeated in many vertebrate radiations, whereby divergence by colouration or other display traits is preceded by specialization to environmental adaptive peaks. PMID- 17465923 TI - Sounds different: inbreeding depression in sexually selected traits in the cricket Teleogryllus commodus. AB - If male sexual signalling is honest because it captures genetic variation in condition then traits that are important mate choice cues should be disproportionately affected by inbreeding relative to other traits. To test this, we investigated the effect of brother-sister mating on advertisement calling by male field crickets Teleogryllus commodus. We quantified the effect of one generation of inbreeding on nightly calling effort and five finer-scale aspects of call structure that have been shown to influence attractiveness. We also quantified inbreeding depression on six life history traits and one morphological trait. Inbreeding significantly reduced hatching success, nymph survival and adult lifespan but had no detectable effect on hatching rate, developmental rate or adult body mass. The effect of inbreeding on sexually selected traits was equivocal. There was no decline in calling effort (seconds of sound production/night) by inbred males, but there were highly significant changes in three of five finer-scale call parameters. Sexually selected traits clearly vary in their susceptibility to inbreeding depression. PMID- 17465924 TI - Geographic structure in a widespread plant-mycorrhizal interaction: pines and false truffles. AB - Mutualistic interactions are likely to exhibit a strong geographic mosaic in their coevolutionary dynamics, but the structure of geographic variation in these interactions is much more poorly characterized than in host-parasite interactions. We used a cross-inoculation experiment to characterize the scales and patterns at which geographic structure has evolved in an interaction between three pine species and one ectomycorrhizal fungus species along the west coast of North America. We found substantial and contrasting patterns of geographic interaction structure for the plants and fungi. The fungi exhibited a clinal pattern of local adaptation to their host plants across the geographic range of three coastal pines. In contrast, plant growth parameters were unaffected by fungal variation, but varied among plant populations and species. Both plant and fungal performance measures varied strongly with latitude. This set of results indicates that in such widespread species interactions, interacting species may evolve asymmetrically in a geographic mosaic because of differing evolutionary responses to clinally varying biotic and abiotic factors. PMID- 17465925 TI - Cope's Rule in the Pterosauria, and differing perceptions of Cope's Rule at different taxonomic levels. AB - The remarkable extinct flying reptiles, the pterosaurs, show increasing body size over 100 million years of the Late Jurassic and Cretaceous, and this seems to be a rare example of a driven trend to large size (Cope's Rule). The size increases continue throughout the long time span, and small forms disappear as larger pterosaurs evolve. Mean wingspan increases through time. Examining for Cope's Rule at a variety of taxonomic levels reveals varying trends within the Pterosauria as a whole, as pterodactyloid pterosaurs increase in size at all levels of examination, but rhamphorhynchoid pterosaurs show both size increase and size decrease in different analyses. These results suggest that analyses testing for Cope's Rule at a single taxonomic level may give misleading results. PMID- 17465926 TI - Complex phenotype-environment associations revealed in an East African cyprinid. AB - Environmental factors influence phenotypes directly, as well as indirectly via trait correlations and interactions with other environmental variables. Using nine populations of the African cyprinid Barbus neumayeri, we employed path analysis to examine direct, indirect and total effects of two environmental variables, water flow (WF) and dissolved oxygen (DO), on several morphological traits. WF and DO directly influenced relative gill size, body shape and caudal fin shape in manners consistent with a priori predictions. Indirect effects also played an important role in the system: (1) strong, oppositely signed direct and indirect effects of WF on body shape resulted in a nonsignificant total effect; (2) DO had no direct effect on body shape, but a strong total effect via indirect effects on gill size; (3) WF indirectly influenced gill size via effects on DO. Only through examination of multiple environmental parameters and multiple traits can we hope to understand complex relationships between environment and phenotype. PMID- 17465927 TI - Recombination is suppressed and variability reduced in a nascent Y chromosome. AB - Several hypotheses have been elaborated to account for the evolutionary decay commonly observed in full-fledged Y chromosomes. Enhanced drift, background selection and selective sweeps, which are expected to result from reduced recombination, may all share responsibilities in the initial decay of proto-Y chromosomes, but little empirical information has been gathered so far. Here we take advantage of three markers that amplify on both of the morphologically undifferentiated sex chromosomes of the European tree frog (Hyla arborea) to show that recombination is suppressed in males (the heterogametic sex) but not in females. Accordingly, genetic variability is reduced on the Y, but in a way that can be accounted for by merely the number of chromosome copies per breeding pair, without the need to invoke background selection or selective sweeps. PMID- 17465928 TI - Life cycle abbreviation in the trematode Coitocaecum parvum: can parasites adjust to variable conditions? AB - The complex life cycles of parasites are thought to have evolved from simple one host cycles by incorporating new hosts. Nevertheless, complex developmental routes present parasites with a sequence of highly unlikely transmission events in order to complete their life cycles. Some trematodes like Coitocaecum parvum use facultative life cycle abbreviation to counter the odds of trophic transmission to the definitive host. Parasites adopting life cycle truncation possess the ability to reproduce within their intermediate host, using progenesis, without the need to reach the definitive host. Usually, both abbreviated and normal life cycles are observed in the same population of parasites. Here, we demonstrate experimentally that C. parvum can modulate its development in its amphipod intermediate host and adopt either the abbreviated or the normal life cycle depending on current transmission opportunities or the degree of intra-host competition among individual parasites. In the presence of cues from its predatory definitive host, the parasite is significantly less likely to adopt progenesis than in the absence of such cues. An intermediate response is obtained when the parasites are exposed to cues from non-host predators. The adoption of progenesis is less likely, however, when two parasites share the resource-limited intermediate host. These results show that parasites with complex developmental routes have transmission strategies and perception abilities that are more sophisticated than previously thought. PMID- 17465929 TI - Viability effects and not meoitic drive cause dramatic departures from Mendelian inheritance for malic enzyme in hybrids of Tigriopus californicus populations. AB - The genetic basis of post-zygotic reproductive isolation is beginning to be untangled in closely related species, but less is known about the genetics of reproductive isolation between divergent populations. Here, two genes encoding malic enzyme (ME) are isolated from the copepod Tigriopus californicus and their influence upon lowered viability in F(2) hybrids of genetically divergent populations is determined. Each ME gene has diverged extensively between T. californicus populations and one gene shows evidence for a recent selective sweep. Segregation patterns of genotypes for both ME genes in adult F(2) hybrids reveal dramatic departures from Mendelian inheritance, deviations that are not seen in F(2) nauplii implying that selection is acting during development based upon the genotype at these ME genes. These results imply that selection against deleterious gene combinations and not aberrant segregation (i.e. meiotic drive) is likely to lead to dramatic departures from Mendelian inheritance observed in these crosses. PMID- 17465930 TI - Short- and long-term benefits and detriments to recombination under antagonistic coevolution. AB - We explored the evolution of recombination under antagonistic coevolution, concentrating on the equilibrium frequencies of modifier alleles causing recombination in initially nonrecombining populations. We found that the equilibrium level of recombination in the host depended not only on parasite virulence, but also on the strength of the modifier allele, and on whether or not the modifier was physically linked to the parasite interaction loci. Nonetheless, the maximum level of recombination for linked loci at equilibrium was about 0.3 (60% of free recombination) for interactions with highly virulent parasites; the level decreased for unlinked modifiers, and for lower levels of parasite virulence. We conclude that recombination spreads because it provides a combination of an immediate (next-generation) fitness benefit and a delayed (two or more generations) increase in the rate of response to directional selection. The relative impact of these two mechanisms depends on the virulence of parasites early in the spread of the modifier, but a trade-off between the two dictates the equilibrium modifier frequency for all nonzero virulences that we examined. In addition, population mean fitness was higher in populations at intermediate equilibria than populations fixed for free recombination or no recombination. The difference, however, was not enough on its own to overcome the two-fold cost of producing males. PMID- 17465931 TI - Do cuckoos choose nests of great reed warblers on the basis of host egg appearance? AB - Prevailing theory assumes cuckoos lay at random among host nests within a population, although it has been suggested that cuckoos could choose large nests and relatively active pairs within host populations. We tested the hypothesis that egg matching could be improved by cuckoos choosing nests in which host eggs more closely match their own, by assessing matching and monitoring nest fate in great reed warblers naturally or experimentally parasitized by eggs of European cuckoos. A positive correlation between cuckoo and host egg visual features suggests that cuckoos do not lay at random within a population, but choose nests and this improves egg matching: naturally parasitized cuckoo eggs were more similar to host eggs as perceived by humans and as measured by spectrophotometry. Our results suggest a hitherto overlooked step in cuckoo-host evolutionary arms races, and have nontrivial implications for the common experimental practice of artificially parasitizing clutches. PMID- 17465932 TI - Compact genes are highly expressed in the moss Physcomitrella patens. AB - Compact genes contain short and few introns, and they are highly expressed in different animal genomes. Recently, it has been shown that in Oryza sativa and Arabidopsis thaliana, highly expressed genes tend to be least compact, containing long and many introns. It has been suggested that selection on genome organization may have acted differently in plants compared with animals. Gene expression can be estimated as the number of hits when comparing a gene sequence with publicly available expressed sequence tags. Here it is shown that in the haploid moss Physcomitrella pates, highly expressed genes contain shorter introns than genes with low expression levels. This study therefore supports the hypothesis that selection may strongly favour transcriptional efficiency at least in the haploid phase of plant life cycles. It is concluded that plants do not necessarily respond to other selection pressures than animals regarding genome structuring. PMID- 17465933 TI - A morphospace-based test for competitive exclusion among flying vertebrates: did birds, bats and pterosaurs get in each other's space? AB - Three vertebrate groups - birds, bats and pterosaurs - have evolved flapping flight over the past 200 million years. This innovation allowed each clade access to new ecological opportunities, but did the diversification of one of these groups inhibit the evolutionary radiation of any of the others? A related question is whether having the wing attached to the hindlimbs in bats and pterosaurs constrained their morphological diversity relative to birds. Fore- and hindlimb measurements from 894 specimens were used to construct a morphospace to assess morphological overlap and range, a possible indicator of competition, among the three clades. Neither birds nor bats entered pterosaur morphospace across the Cretaceous-Paleogene (Tertiary) extinction. Bats plot in a separate area from birds, and have a significantly smaller morphological range than either birds or pterosaurs. On the basis of these results, competitive exclusion among the three groups is not supported. PMID- 17465934 TI - Inbreeding avoidance in spiders: evidence for rescue effect in fecundity of female spiders with outbreeding opportunity. AB - Selection by inbreeding depression should favour mating biases that reduce the risk of fertilization by related mates. However, equivocal evidence for inbreeding avoidance questions the strength of inbreeding depression as a selective force in the evolution of mating biases. Lack of inbreeding avoidance can be because of low risk of inbreeding, variation in tolerance to inbreeding or high costs of outbreeding. We examined the relationship between inbreeding depression and inbreeding avoidance adaptations under two levels of inbreeding in the spider Oedothorax apicatus, asking whether preference for unrelated sperm via pre- and/or post-copulatory mechanisms could restore female fitness when inbreeding depression increases. Using inbred isofemale lines we provided female spiders with one or two male spiders of different relatedness in five combinations: one male sib; one male nonsib; two male sibs; two male nonsibs; one male sib and one male nonsib. We assessed the effect of mating treatment on fecundity and hatching success of eggs after one and three generations of inbreeding. Inbreeding depression in F1 was not sufficient to detect inbreeding avoidance. In F3, inbreeding depression caused a major decline in fecundity and hatching rates of eggs. This effect was mitigated by complete recovery in fecundity in the sib-nonsib treatment, whereas no rescue effect was detected in the hatching success of eggs. The rescue effect is best explained by post-mating discrimination against kin via differential allocation of resources. The natural history of O. apicatus suggests that the costs of outbreeding may be low which combined with high costs of inbreeding should select for avoidance mechanisms. Direct benefits of post-mating inbreeding avoidance and possibly low costs of female multiple mating can favour polyandry as an inbreeding avoidance mechanism. PMID- 17465935 TI - Transfusion medicine illustrated: B subgroup mistyped as O. PMID- 17465936 TI - Optimizing the use of limited rare red cells for hematopoietic cell transplant recipients. PMID- 17465937 TI - Safer design. PMID- 17465938 TI - How do I manage a blood shortage in a transfusion service? PMID- 17465939 TI - Radiofrequency identification technology can standardize and document blood collections and transfusions. AB - BACKGROUND: We evaluated the potential for radiofrequency (RF) transponder microchips to standardize and document key steps in the blood collection and transfusion process. STUDY DESIGN AND METHODS: Using the blood center's standard operating procedures for blood collections, we programmed a laptop computer and 10 multiwrite 256-byte RF microchips to prompt operators to enter data for key steps in blood collection. Before collections, RF microchips were attached to blood collection sets at the blood center. In parallel with actual collections, we added data to the microchips with the computer and a hand-held scanner programmer. After labeling, we shipped the RF microchip-tagged blood units to the hospital where unit-related data (whole blood number, ABO and Rh, expiration date, special laboratory test results) were uploaded from the RF microchip to the transfusion service's information system. The microchip was subsequently used as a cross-match label for blood unit-recipient matching. RESULTS: Data were successfully uploaded to the RF microchip at key steps during blood collections. Software programs in the laptop computer and hand-held scanner-programmer successfully prompted operators to enter key data. At any stage in a blood collection, authorized operators were able to review electronic records of prior steps using the laptop computer or by scanning the microchip attached to the blood bag. Unit-related data were successfully transferred to the hospital transfusion service through the RF microchip. These data were successfully incorporated in the RF microchip cross-match label, which was used to confirm recipient-blood unit matching at the bedside. CONCLUSION: RF microchips can collect key data during blood collections, facilitate information transfer from the blood center to the hospital, and confirm recipient-blood unit matching at the bedside before transfusions. PMID- 17465940 TI - Prevention of bedside errors in transfusion medicine (PROBE-TM) study: a cluster randomized, matched-paired clinical areas trial of a simple intervention to reduce errors in the pretransfusion bedside check. AB - BACKGROUND: Transfusion of the incorrect blood component is a frequent serious incident associated with transfusion and often involves misidentification of the patient and/or the unit of blood. The objective of this study was to assess the effect of a simple intervention designed to improve performance of the bedside check and to observe the durability of any effect. The intervention was a tag on blood bags reminding staff to check the patient's wristband. The tag was positioned in such a way that the transfusionist was required to remove the tag to spike the unit. STUDY DESIGN AND METHODS: The intervention was tested in a multicenter cluster-randomized controlled trial incorporating short-term and long term follow-up periods. The primary endpoint was the proportion of patients transfused with red cell units for whom the key elements of the bedside check were all correctly completed. RESULTS: Fifteen matched-paired clinical areas at 12 participating hospitals in six countries were included in the trial. Combining data from all participating hospitals, the bedside check was correctly performed in 37 percent of transfusions during the baseline audit period. There was no evidence of a favorable effect of the intervention immediately after its introduction (pooled odds ratio, 1.09; 95% confidence interval, 0.54-2.17). There was similarly no evidence of a favorable effect after continued use of the intervention for an additional 8 weeks. CONCLUSIONS: A simple intervention in the form of a barrier warning label on blood bags reminding staff to check the patient's wristband failed to improve bedside transfusion practice. The robust study design developed for this study could be applied to investigate other interventions to improve the safety of bedside transfusion practice. PMID- 17465941 TI - Transfusion policy: when to stop the use of extremely rare blood for an allogeneic hematopoietic progenitor cell transplant recipient with a history of red cell alloimmunization. AB - BACKGROUND: Decisions for when to select, and when to discontinue, antigen negative blood in hematopoietic progenitor cell transplantation (HPCT) recipients with red blood cell (RBC) antibodies can be confusing. In HPCT performed for sickle cell anemia patients who require extremely rare antigen-negative blood, the balance of caution and practicality is further complicated. CASE REPORTS: Four sickle cell anemia patients with current or historic RBC antibodies underwent allogeneic HPC transplantation. One required extremely rare (group O D , hr(B)-) blood. None of the antibodies caused significant hemolysis after transplant. In the case requiring rare blood, antigen-negative blood was requested after donor RBC engraftment because of incomplete donor white blood cell (WBC) chimerism. CONCLUSIONS: RBC antibodies derived from a recipient of allogeneic HPCT rarely cause significant hemolysis, in contrast to the more severe picture sometimes seen with donor-derived antibodies. When donor WBC chimerism is delayed past the time of donor RBC engraftment, there can be concern for the possibility of future recipient-type antibody production. Even 100 percent donor lymphocyte chimerism is no guarantee of total host plasma cell ablation. Immunoglobulin allotyping, when informative, can suggest chimerism for several years. Recipient-type blood, when extremely rare, may not be available for that duration. PMID- 17465942 TI - False-positive pregnancy test after passive transfusion of beta-human chorionic gonadotropin from donor red blood cells during erythrocytapheresis. AB - BACKGROUND: Patients transfused with blood products may passively receive soluble antibodies, proteins, and other analytes that persist during the collection, processing, and transfusion of the blood product. In this report, a female patient who received transfusion of five red blood cell (RBC) units during erythrocytapheresis later demonstrated an unexpected positive result in assays for the beta-subunit of human chorionic gonadotropin (bHCG), a screening test for pregnancy. The result caused postponement of an elective surgical procedure. A follow-up test 1 week later was negative. STUDY DESIGN AND METHODS: To investigate the possibility of passive transfusion of the hormone from a donor RBC unit, a sample from each of the units transfused was assayed for the level of bHCG. RESULTS: One of the 5 units transfused to the patient had a high level of bHCG. The observed bHCG level in the recipient was found to be comparable to the predicted level, given the donor's plasma bHCG level and accounting for the dilution factors in the preparation of the RBC unit and the erythrocytapheresis procedure and the in vivo t((1/2)) of the hormone. CONCLUSION: The donor, who was unaware of her pregnancy status at the time of donation, harbored a high bHCG level that caused the positive test result in the recipient patient's serum and urine. If an unexpected analyte or serology is detected in a recipient of a blood transfusion, it is important to consider the possibility of passive transfusion of the analyte. PMID- 17465943 TI - Thalassemia intermedia today: should patients regularly receive transfusions? AB - BACKGROUND: beta-Thalassemia is an inherited hemoglobin disorder characterized by reduced synthesis of beta-globin chain. The severity of clinical course distinguishes this heterogeneous disease in two main subtypes: thalassemia major (TM) and thalassemia intermedia (TI). TI has a later clinical onset with a milder anemia that does not require transfusions at least during the first few years of life. The clinical picture of TI patients who have not received transfusions or have occasionally received transfusions is dominated by the consequences of chronic hemolytic anemia, tissue hypoxia, and their compensatory reactions, such as bone deformities and fractures, extramedullary hemopoiesis, spleen and liver enlargement, hypercoagulability, and pulmonary hypertension. These complications, especially the latter two, are getting more frequent and severe over the years. Nowadays, although TI patients have almost no changes in the course of the disease, well-treated TM patients with regular transfusion-chelation therapy showed suppression of the anemia-related disorders in parallel to prolongation of life. The new oral iron chelators and the magnetic resonance imaging application for early detection of heart iron load are promising for further improvement on survival. CONCLUSIONS: Considering the current cost-benefit balance of regular treatment in TM as well as the frequency and severity of complications in TI, it seems that the majority of TI patients will be benefited if this kind of treatment is applied targeting prevention and not palliation of the anemia induced complications. PMID- 17465944 TI - Acute bleeding complications in patients after hematopoietic stem cell transplantation with prophylactic platelet transfusion triggers of 10 x 10(9) and 20 x 10(9) per L. AB - BACKGROUND: Prophylactic platelet (PLT) transfusions are given as a standard care in patients with hematologic malignancies undergoing hematopoietic stem cell transplantation (HSCT). This retrospective analysis evaluates utilization of blood transfusions, risk of bleeding, and survival in 480 HSCT patients at 10 x 10(9) and 20 x 10(9) per L prophylactic trigger levels. STUDY DESIGN AND METHODS: A total of 224 patients received prophylactic PLT transfusions at 20 x 10(9) per L threshold (1997-1998, SP1); 256 patients had prophylaxis at 10 x 10(9) per L (1999-2001, SP2). Bleeding scores were assigned daily. RESULTS: A slight reduction in PLT transfusions per patient in SP2 compared with SP1 was not statistically significant (odds ratio, 0.82; 95% confidence interval, 0.51-1.33; p = 0.416), yet a significantly higher proportion of patients in SP2 had PLT counts less than or equal to 10 x 10(9) per L compared to SP1 (p < 0.001). In patients who bled, however, there was no excess exposure to low PLT counts before bleeding started. A substantial number of patients who bled received PLT transfusions above the goal before bleeding started (82.9% in SP2, 41.5% in SP1) because of medical complications that associated with increased risk of bleeding. Bleeding incidence was similar in both study periods (21.9% in SP1, 16.4% in SP2; p = 0.526). Bleeding was significantly associated with reduced survival in both study periods. CONCLUSIONS: Patients who bled were usually placed on a higher threshold before the onset of their major bleeding event and were not exposed to additional risk of bleeding from thrombocytopenia. Similarity in bleeding incidence between study periods appears to associate with adjustments to high risk conditions and may not reflect consequences of the lower transfusion threshold. PMID- 17465945 TI - Coexistence of autoantibodies and alloantibodies to red blood cells due to blood transfusion. AB - BACKGROUND: Autoantibodies (AUTO) to red blood cells (RBCs) are frequently associated with alloantibodies (ALLO). The mechanism for the coexistence of these antibodies is obscure. STUDY DESIGN AND METHODS: Between August 1998 and June 2006, all in- and outpatients of the Charite University Hospital, Berlin, with detectable AUTO were included in this study. Serologic examination was performed with standard techniques for the detection of RBC antibodies. RESULTS: A total of 717 patients were found to have AUTO, with ALLO observed in 200 of these patients (28%). The history of antibody production could be evaluated in 98 of the 200 patients. Both AUTO and ALLO were due to RBC transfusion in 73 cases (75%) and peripheral blood progenitor cell transplantation in 6 cases (6%). Nine (9%) patients were primarily alloimmunized and subsequently developed AUTO. The remaining 10 (10%) patients were primarily autoimmunized and later developed ALLO. Only 6 of these patients had chronic idiopathic autoimmune hemolytic anemia. CONCLUSION: The majority of AUTO associated with ALLO appears to be due to RBC transfusion that must be recognized as a major cause for autoimmunization. PMID- 17465946 TI - Prediction of engraftment after autologous peripheral blood progenitor cell transplantation: CD34, colony-forming unit-granulocyte-macrophage, or both? AB - BACKGROUND: The rate of hematologic recovery after peripheral blood progenitor cell (PBPC) transplantation is influenced by the dose of progenitor cells. Enumeration of cells that express CD34+ on their surface is the most frequently used method to determine progenitor cell dose. In vitro growth of myeloid progenitor cells (colony-forming unit-granulocyte-macrophage [CFU-GM]) requires more time and resources, but may add predictive information. STUDY DESIGN AND METHODS: A series of 323 patients, who underwent autologous PBPC transplantation for multiple myeloma, malignant lymphoma, or locally advanced breast cancer, were studied for the effect of CD34+ dose and CFU-GM dose on hematologic recovery. Measures for engraftment were days to absolute granulocyte and platelet (PLT) counts to greater than 500 per muL and than 20 x 10(9) per L, respectively, and number of PLT transfusions and red cell units required. RESULTS: The CD34+ dose had a median of 8.4 x 10(6) per kg, and the CFU-GM dose a median of 84.9 x 10(4) per kg. The CD34+ and CFU-GM doses showed significant correlation (R = 0.63; p < 0.0001) but a wide variation in the ratio of CD34+ and CFU-GM. Both CD34+ and CFU GM doses had significant correlation with the measures of engraftment, but for all measures the relationship of CD34+ was stronger. Multivariate analysis and subgroup analysis of patients receiving CD34+ doses of less than 5 x 10(6) per kg also did not reveal an independent predictive value for CFU-GM. CONCLUSION: For prediction of hematologic recovery after autologous PBPC transplantation, determination of CFU-GM dose does not add to the predictive value of the CD34+ dose. PMID- 17465947 TI - Evaluation of the COM.TEC cell separator in predicting the yield of harvested CD34+ cells. AB - BACKGROUND: This multicenter study was performed with the intention to evaluate the exactness of the predicted CD34+ cell yield calculated by two leukapheresis programs of the cell separator COM.TEC upon the number of donor's circulating CD34+ cells and the blood volume processed. STUDY DESIGN AND METHODS: Patients and healthy donors (n = 166) received mobilization by chemotherapy and/or granulocyte colony-stimulating factor and underwent CD34+ cell harvest by the leukapheresis programs MNC or RV-PBSC (n = 203). RESULTS: CD34+ cells were collected by 112 harvests on MNC and by 91 collections on RV-PBSC. The median collection efficiency of CD34+ cells was significantly better for the program MNC than for RV-PBSC (p < 0.001): 67% (31-109) vs. 42% (19-100). The collected CD34+ cell yield was in median more exactly by MNC than by RV-PBSC (p < 0.001): 85% (31 176) vs. 59% (22-110) of the predicted value. Concentrates obtained by RV-PBSC showed in median significantly higher percentages of mononuclear cells (p < 0.001) and CD34+ cells (p < 0.001), 86% (43-99) vs. 56% (25-95) and 1.2% (0.2 14.3) vs. 0.4% (0.1-6.0), and had lower contaminations by erythrocytes (p < 0.001) and platelets (p < 0.001), 13 mL (4-48) vs. 25 mL (5-60) and 1.9 x 1011 vs. 3.1 x 1011, than those harvested by MNC. CONCLUSION: The significantly better collection efficiency of CD34+ cells and the more exact prediction of the harvested CD34+ cell yield make the leukapheresis program MNC a safe and efficient procedure. However, concentrates collected by RV-PBSC are of a better cellular quality with a significantly higher percentage of mononuclear and CD34+ cells and a lower contamination by erythrocytes and platelets. PMID- 17465948 TI - A cluster-randomized controlled trial of a blood conservation algorithm in patients undergoing total hip joint arthroplasty. AB - BACKGROUND: The optimum strategy for reducing allogeneic blood transfusion in patients undergoing total hip joint arthroplasty (THJA) is unknown. STUDY DESIGN AND METHODS: The effectiveness of a comprehensive blood conservation algorithm (BCA) was evaluated by means of a cluster randomization trial. Thirty hospitals performing primary THJA were randomly assigned to implement the algorithm or to continue with usual care (UC). Subsequently, the institutional rate of allogeneic transfusion was determined for 60 consecutive patients who underwent surgery at each site. The BCA consisted of patient and provider education, hemoglobin-based recommendations for specific blood conservation strategies (recombinant human erythropoietin [rHuEPO] or autologous blood donation [ABD]) and transfusion guidelines. The main outcome measure was the institutional allogeneic transfusion rate. RESULTS: One hospital withdrew consent after randomization, resulting in 14 hospitals assigned to BCA and 15 to UC. In the BCA arm, the institutional rates of rHuEPO use and ABD participation were 20.1 and 27.1 percent compared to 0.6 and 25.8 percent, respectively, in the UC arm. The allogeneic transfusion rate was substantially reduced in hospitals assigned to the BCA group (p = 0.02; absolute risk reduction, 9.6% [26.1% UC vs. 16.5% BCA]). Multivariate analysis of patient-level data showed that assignment to the UC arm was an independent risk factor for allogeneic transfusion (p = 0.037; odds ratio, 1.8; 95% confidence interval, 1.0-3.1) when adjusted for other prognostic factors. No differences were observed in the use of autologous blood. CONCLUSION: A comprehensive approach to blood conservation was superior to UC for reducing allogeneic transfusion in patients undergoing THJA. PMID- 17465949 TI - Identification and characterization of a novel antisense RNA transcribed from the opposite strand of the human blood group ABO gene. AB - BACKGROUND: To elucidate the molecular basis of control of the ABO gene in cell type-specific expression, during normal cell differentiation, and in cancer cells lacking A/B antigen, the mechanisms responsible for regulation of human ABO gene expression have been studied. Recently, naturally occurring antisense transcriptions have been reported to regulate gene expression through a variety of biological mechanisms. Therefore, RNA transcribed from the opposite strand of the ABO gene was investigated. STUDY DESIGN AND METHODS: The presence of antisense RNA to the ABO-coding strand in human cancer cell lines and normal tissues was examined by strand-specific reverse transcription-polymerase chain reaction. The 5'- and 3'-ends of the transcript were determined by the rapid amplification of cDNA ends (RACE) system. KATOIII cells were treated with mithramycin A, followed by quantitative analysis of both sense and antisense ABO transcripts. RESULTS: The endogenous antisense RNA to the ABO coding strand was found to start within the first intron of the ABO gene, and the expression coincided with ABO gene expression in various cultured cells and normal tissues. This novel gene was named ABOAS. Treatment of KATOIII cells with mithramycin A repressed transcription from the ABO exon 1 promoter, while it increased the ABOAS transcript. CONCLUSION: These results suggest that ABOAS transcribed from the opposite strand of the ABO gene might be involved in the regulation of ABO gene expression. PMID- 17465950 TI - A new DEL variant caused by exon 8 deletion. AB - BACKGROUND: A 28-year-old woman of Lebanese origin experienced two stillbirths. At the time, serology typed her red blood cells as being group A D- and found an anti-D in her serum sample. Molecular biology analysis, however, showed that she was in fact RHD+. STUDY DESIGN AND METHODS: To better characterize this case, a full investigation including family members was undertaken. Classical serology techniques and DNA and RNA analysis were performed whenever possible. RESULTS: Serology results showed that the patient's father and two brothers were D-. RHD genotyping demonstrated that her two brothers were indeed RHD+. Polymerase chain reaction (PCR) amplification was performed on each RHD 10 exons. Exon 8 did not amplify for the patient, her father, and her two brothers. Her mother and sister had exon 8. Messenger RNA analysis showed five RHD transcripts. The longest transcript was missing exon 8 but had a part of intron 7 inserted instead. Genomic DNA sequencing revealed a 995-bp deletion including part of intron 7, exon 8, and intron 8. This mutation, RHD(delEx8), was found to express a DEL in adsorption-elution. To facilitate the screening of this new DEL allele, a simple PCR-based assay was designed. CONCLUSION: This novel allele represents the first observation of a large deletion at the genomic level within the RHD gene in Caucasian persons. PMID- 17465951 TI - Identification of 12 novel RHD alleles in western France by denaturing high performance liquid chromatography analysis. AB - BACKGROUND: Unlike the standard RHD+ or RHD- alleles, serologic determination of weak or partial D alleles is often not clear-cut. Most importantly, rare weak D alleles, not typed by serology, are prone to alloimmunization when transfused with D+ blood. Although more than 100 RHD variants have currently been reported, many more rare alleles probably remain to be identified. STUDY DESIGN AND METHODS: To identify novel unusual RHD alleles, genomic DNA samples were collected from 333 blood donors or recipients in western France. All displayed ambiguity for D phenotype as determined by routinely used serologic reagents and analyzed by means of denaturing high-performance liquid chromatography (DHPLC) analysis in parallel with direct sequencing. RESULTS: For the first time it has been established that a reliable DHPLC-based approach potentiates the rapid screening of the entire RHD gene-coding sequence. In so doing, a total of 12 novel RHD alleles were identified. Except for the null allele that is in trans with a Weak D type 4 allele, the predicted effects of the other new alleles on gene expression correlated well with the discrepant routine D phenotype results. In particular, the carrier of the p.Leu214Phe missense mutation developed alloanti-D antibodies after transfusion of D+ blood. CONCLUSION: The identification of 12 novel RHD alleles represents a significant addition to the known repertoire of unusual RHD variants and, at the same time, serves to deepen our understanding of the molecular basis of weak and partial D. The accurate molecular typing of RHD alleles would allow to reduce the alloimmunization risk. PMID- 17465952 TI - Structural basis for red cell phenotypic changes in newly identified, naturally occurring subgroup mutants of the human blood group B glycosyltransferase. AB - BACKGROUND: Four amino-acid-changing polymorphisms differentiate the blood group A and B alleles. Multiple missense mutations are associated with weak expression of A and B antigens but the structural changes causing subgroups have not been studied. STUDY DESIGN AND METHODS: Individuals or families having serologically weak B antigen on their red cells were studied. Alleles were characterized by sequencing of exons 1 through 7 in the ABO gene. Single crystal X-ray diffraction, three-dimensional-structure molecular modeling, and enzyme kinetics showed the effects of the B allele mutations on the glycosyltransferases. RESULTS: Seven unrelated individuals with weak B phenotypes possessed seven different B alleles, five of which are new and result in substitution of highly conserved amino acids: M189V, I192T, F216I, D262N, and A268T. One of these (F216I) was due to a hybrid allele resulting from recombination between B and O(1v) alleles. The two other alleles were recently described in other ethnic groups and result in V175M and L232P. The first crystal-structure determination (A268T) of a subgroup glycosyltransferase and molecular modeling (F216I, D262N, L232P) indicated conformational changes in the enzyme that could explain the diminished enzyme activity. The effect of three mutations could not be visualized since they occur in a disordered loop. CONCLUSION: The genetic background for B(w) phenotypes is very heterogeneous but usually arises through seemingly random missense mutations throughout the last ABO exon. The targeted amino acid residues, however, are well conserved during evolution. Based on analysis of the resulting structural changes in the glycosyltransferase, the mutations are likely to disrupt molecular bonds of importance for enzymatic function. PMID- 17465953 TI - Synthetic glycolipid modification of red blood cell membranes. AB - BACKGROUND: Glycolipids have a natural ability to insert into red cell (RBC) membranes. Based on this concept the serology of RBCs modified with synthetic analogs of blood group glycolipids (KODE technology) was developed, which entails making synthetic glycolipid constructs engineered to have specific performance criteria. Such synthetic constructs can be made to express a potentially unlimited range of carbohydrate blood group determinants. STUDY DESIGN AND METHODS: Synthetic constructs incorporating A, B, acquired-B, and Le(a) blood group determinants were constructed and used to modify RBCs. Modified cells were assessed by routine serologic methods using a range of commercially available monoclonal antibodies. RESULTS: RBCs modified with different concentrations of synthetic glycolipids were able to give controllable serologic results. Synthetic A and B modified cells were able to be created to represent the serology of "weak" subgroups. Specialized cells such as those bearing synthetic acquired-B antigen reacted as expected, but also exhibited extended features due to the cells bearing only specific antigen. Synthetic Le(a)-modified cells reacted as expected with anti-Le(a) reagents, but unexpectedly, were also able to detect the chemical anti-Le(ab) specificity of serologic monoclonal anti-Le(b) reagents. CONCLUSION: RBCs can be created to express normal and novel carbohydrate profiles by inserting synthetic glycolipids into them. Such cells will be useful in creating specialized antigen panels and for quality control purposes. PMID- 17465954 TI - Parvovirus B19 DNA in Factor VIII concentrates: effects of manufacturing procedures and B19 screening by nucleic acid testing. AB - BACKGROUND: Parvovirus B19 (B19) is a common contaminant, especially in coagulation factors. Because of B19 transmission by pooled plasma, solvent/detergent treated in 1999, some fractionators initiated minipool nucleic acid testing (NAT) to limit the B19 load in manufacturing pools. In this study, the extent of B19 DNA contamination in commercial Factor VIII concentrates, that is, antihemophilic factor (human) (AHF), manufactured before and after B19 NAT screening was implemented, was determined. STUDY DESIGN AND METHODS: A total of 284 lots representing six AHF products made during 1993 to 1998 and 2001 to 2004 were assayed for B19 DNA by an in-house NAT procedure. Anti-B19 immunoglobulin G (IgG) was also measured. RESULTS: Most lots made during 1993 to 1998 had detectable B19 DNA. The prevalence ranged from 56 to 100 percent and appeared to differ between manufacturers. The highest level of B19 DNA found was 10(6) genome equivalents (geq or international units [IU]) per mL. Forty percent of the lots tested contained 10(3) geq (IU) per mL. In comparison, both prevalence and levels in source plasma-derived AHF products made in 2001 to 2004 were lower. Both, however, remained unchanged in the recovered plasma-derived product because B19 NAT screening had not been implemented. Only an intermediate-purity AHF product was positive for the presence of anti-B19 IgG. CONCLUSION: The prevalence and levels of B19 DNA in AHF prepared from B19 NAT unscreened plasma were high but varied among products with different manufacturing procedures. B19 NAT screening of plasma effectively lowered the B19 DNA level in the final products and in the majority of cases rendered it undetectable and hence potentially reduced the risk of B19 transmission. PMID- 17465955 TI - Seasonal temperature variation and the rate of donor deferral for low hematocrit in the American Red Cross. AB - BACKGROUND: Hematocrit (Hct) values in healthy adult populations exhibit seasonal variation, with the lowest values occurring in the summer. The extent to which environmental temperature contributes to the seasonal trend in deferral rates for unacceptable Hct in the American Red Cross was further analyzed. STUDY DESIGN AND METHODS: A centralized database of donations during 2002 to 2004, constituting 24.3 million donor presentations, was further characterized. Data on mean monthly temperature in the United States were obtained for the same period from a government agency. Multivariate regression analyses were performed to determine the relationship between Hct deferral rates among blood donors and environmental temperature and donor characteristics. RESULTS: Hct deferral rates were associated with mean monthly temperature in the United States (R(2) = 0.77). The relationship between the Hct deferral rate and environmental temperature was strongest in the region of the country with the highest seasonal variation in temperature, followed by regions with intermediate and low seasonal variation in temperature, respectively. The seasonal pattern in Hct deferral rates occurred in both sexes and across all age groups, with significantly higher Hct deferral rates occurring in June through August compared to other quarters (p < 0.0007). CONCLUSION: There is a significant seasonal pattern in Hct deferral rates that is associated with environmental temperature. The relationship between Hct deferral rates and temperature is strongest in areas of the country with greater temperature variability. The effect of seasonality on Hct deferrals should be taken into account for donor counseling, recruitment, and retention efforts. PMID- 17465956 TI - Seasonal influences on hemoglobin levels and deferral rates in whole-blood and plasma donors. AB - BACKGROUND: Indications exist that deferral rates in blood donors are highest in summer. However, a detailed quantitative analysis is not available. The association between Hb values, deferral rates, and daily temperatures was investigated in a large data set of blood donors. STUDY DESIGN AND METHODS: The study population consisted of both plasma and whole-blood donors from the southeast region of the Netherlands. Individual Hb levels and other examination data between January 2002 and December 2004 were extracted from the donor file. Data on daily maximum temperatures were related to Hb levels and Hb deferrals. Results are reported separately for plasma and whole-blood donors as well as for men and women. RESULTS: Data were available from 106,398 whole blood donors and 6983 plasma donors, resulting in data of more than 600,000 examinations. Hb levels decreased with increasing daily temperature. Highest deferral rates were observed in summer months, which were consistent over the several groups and over the three years. The highest Hb deferral of 11.1 percent was observed for female whole-blood donors on days with a maximum temperature of 25 degrees and above. In all four donor categories a gradual increase with temperature was observed with 1.7-2.2 times higher deferral rates on hot days (> or = 25 degrees C) compared to cold days (<5 degrees C). CONCLUSION: A clear seasonal pattern in Hb levels and in the percentage of Hb deferrals was observed. The observed seasonal effect could not be explained by differences in donor characteristics. Our observations might have practical implications for donor management. PMID- 17465957 TI - Management and outcome of 200 cases of fetomaternal alloimmune thrombocytopenia. AB - BACKGROUND: Fetomaternal alloimmune thrombocytopenia (FMAIT) is the commonest cause of severe thrombocytopenia in term neonates but its management remains controversial. STUDY DESIGN AND METHODS: A 7-year prospective observational study of 200 cases of FMAIT evaluated the relationship between human platelet antigen (HPA) antibody specificity, clinical presentation, morbidity, mortality, and therapeutic interventions in the antenatal and postnatal period, with long-term follow-up of neonates with intracranial hemorrhage (ICH). RESULTS: In 1148 referrals for FMAIT, HPA antibodies were confirmed in 200 (17%). The commonest specificities were anti-HPA-1a, 150 (75%); anti-HPA-5b, 31 (15.5%); and anti-HPA 15b, 8 (4%). Of 123 (62%) cases (two sets of twins) with no previous history of FMAIT, intrauterine deaths occurred in 5: anti-HPA-1a alone, 3; in combination with anti-HPA-5b, 1; and anti-HPA-15b, 1. Of the 120 live neonates, 103 had severe thrombocytopenia and 17 (14%) developed ICH (anti-HPA-1a, 13; anti-HPA-5b, 3; anti-HPA-15b, 1). Postnatal care varied widely with 37 percent of neonates receiving random rather than HPA-1a and -5b-negative platelets. Of the remaining 77 cases with a history of FMAIT, 40 received intrauterine transfusions. Six (15%) of these fetuses died in utero and an additional 2 developed ICH postnatally. Of the 19 children with ICH, 1 (anti-HPA-15b) died on Day +1, and neurologic sequelae persist in 13 (mean follow-up, 2.5 years). CONCLUSION: HPA-1a antibodies are most commonly implicated in severe thrombocytopenia but HPA-5b and HPA-15b antibodies can also result in poor outcome. Postnatal transfusion management is extremely variable, and fetal transfusions are associated with significant morbidity and mortality. PMID- 17465958 TI - Fetal hemolytic anemia and intrauterine death caused by anti-M immunization. AB - BACKGROUND: Antibodies with anti-M specificity are detected in 10 percent of pregnant women with a positive antibody screen, but anti-M is only rarely associated with hemolytic anemia in the fetus. STUDY DESIGN AND METHODS: This study reports on three pregnancies in one family that all resulted in severe fetal anemia. The first fetus died in utero with hydrops fetalis during the 20th gestational week and the second child was delivered after 28 weeks of gestation with hydrops fetalis and a hemoglobin level of 16 g per L whereas the third affected child was treated with intrauterine red cell (RBC) transfusions before delivery at 28 weeks of gestation. RESULTS: The direct antiglobulin test was negative but anti-M in a low titer was detected through the three pregnancies, and its clinical relevance, which initially was uncertain, was confirmed by pronounced in vivo hemolysis in maternal blood of chromate ((51)Cr)-labeled M+ RBCs and normal survival of (51)Cr labeled M- RBCs. CONCLUSION: It is concluded that anti-M immunization in a few cases may cause severe fetal hemolytic anemia and intrauterine death. It remains to be elucidated why a normally clinically insignificant antibody is this aggressive in a small proportion of cases. Because the condition is treatable, anti-M must be considered as a possible cause of fetal anemia and intrauterine death. PMID- 17465959 TI - Y-chromosome short tandem repeats analysis to complement paternal lineage study: a single institutional experience in Taiwan. AB - BACKGROUND: Highly polymorphic autosomal short-tandem-repeat (STR) analysis can be useful in most kinship testing. Y-chromosome-specific STRs, in contrast, have been increasingly applied for the verification of equivocal paternal genetic transmissions. STUDY DESIGN AND METHODS: A total of 338 unrelated males were first typed for the 9-loci Y-STR European minimal haplotype (minHt). Samples with haplotypes that were found at least two times were subject to further study by a commercially available 17-Y-STR multiplex set (AmpFlSTR Yfiler). A separate clinical study for 113 various kinship identifications of male genetic transmission were then conducted by a panel consisting of 18 autosomal STRs and complemented by both Y-STR multiplex sets and their respective results compared. RESULTS: For the 338 individuals, a total of 270 haplotypes were identified after the minHt study, of which 234 were unique. Among the rest of the 104 samples, AmpFlSTR Yfiler identified 82 other unique haplotypes. Altogether, 324 different haplotypes were observed; 316 (97.5%) were unique whereas 8 were shared by two to seven times. The haplotype diversities for the minHt and the AmpFlSTR Yfiler were 99.75 and 99.96 percent, respectively, whereas the powers of discrimination (PDs) were 79.88 and 95.86 percent, respectively. Despite a lower PD for minHt, there was no discrepancy on the clinical setting for personal identification between the two Y-STR sets in an allegedly true male lineage transmission involving 66 cases with 24 father-son, 19 siblings, 9 uncle-nephew, 8 grandfather-grandson, 3 cousins, and 3 half-siblings. For 47 other cases with a false allegation of paternity, exclusion was made for all without ambiguity by either Y-STR panel. CONCLUSION: The 9-loci minHt Y-STR set is adequate to complement conventional autosomal STRs for kinship studies where Y-lineage transmission is concerned. PMID- 17465960 TI - Managing a tissue recall in a large academic hospital. AB - BACKGROUND: In October 2005, a recall of human tissue for implantation was initiated because one recovery center obtained tissues from donors who were not screened properly for infectious diseases. The Food and Drug Administration (FDA) and Centers for Disease Control (CDC) recommended notifying affected patients and offering access to infectious disease testing. STUDY DESIGN AND METHODS: A multidisciplinary team was established to provide a framework for responding to the recall. The plan was designed to meet six goals. Steps included patient identification, surgeon and patient notification, patient education and testing, communication of test results, and information for the public. RESULTS: The institution received 55 recalled tissues, of which 48 had been implanted into 30 patients undergoing neurosurgical, orthopedic, and general surgical procedures. Patients were identified and sent notification letters within 2 weeks of the FDA and CDC recommendations. Twenty-seven patients underwent testing, which was performed at the convenience of the patients at no cost to them. One patient had evidence of previous (but not current) hepatitis B infection. Overall, patients were appreciative of the processes used. Media coverage was positive. CONCLUSION: The response plan was generally successful in achieving the established goals. Potential improvements were identified in several areas, including initial patient notification and coordination of test result communication. It is critical to allow flexibility to meet each patient's needs. The plan may serve as a template for use in future tissue recalls by other hospital-based tissue banks. PMID- 17465961 TI - Cryopreservation of living cells: principles and practice. AB - Increasingly, the cryopreservation of living cells is being attempted by researchers whose primary interest and experience is with the medical applications of those cells or tissues and whose prior experience with cryobiology may be negligible. It is therefore generally necessary to imitate some regimen used by others, perhaps with some other cell type and attempt to optimize the recovery empirically. This article makes no attempt to provide specific protocols for the many individual cell types. Rather it is a primer that may help to give such investigators an insight into the basic principles of cell freezing, cryoprotectants, and, particularly, their addition and removal. Finally, the article summarizes the five different approaches to applied cryopreservation: ultrarapid freezing and thawing, controlled-rate freezing, freezing with nonpenetrating polymers, vitrification, and equilibrium freezing. PMID- 17465962 TI - Platelet radiolabeling procedure. PMID- 17465963 TI - Paternity inclusions among suspicious spouses and their social value. PMID- 17465965 TI - Anti-Wr(a): to screen or not to screen? PMID- 17465966 TI - A predonation screening question for occupational exposure to simian foamy virus: a preliminary donor survey in Canada. PMID- 17465969 TI - Intravenous indigo carmine might cause cerebral ischemia. PMID- 17465970 TI - Oral naproxen but not oral paracetamol reduces the need for rescue analgesic after adenoidectomy in children. AB - BACKGROUND: Our aim was to show the efficacy of naproxen and paracetamol with and without pethidine on pain and nausea and vomiting after adenoidectomy. The primary outcome was the requirement of rescue analgesic for post-operative pain and the secondary outcome was post-operative nausea and vomiting (PONV). METHODS: A randomized, double-blind, placebo-controlled study design was used. Thirty minutes before anaesthesia induction, patients (n= 180) received either a single oral dose analgesic (naproxen 10 mg/kg or paracetamol 20 mg/kg) or a placebo. Half of the children received pethidine 1 mg/kg intravenously (i.v.) at the induction of anaesthesia. Post-operative pain was evaluated using an objective behavioural pain scale (OPS 0-9) and rescue medication, i.v. fentanyl 1 mug/kg, was administered if the child suffered from moderate or severe pain (OPS > or = 4). RESULTS: When pethidine was not used, 83% of the children in the naproxen group vs. 97% in the other two groups required rescue fentanyl (P < 0.05). The use of pethidine reduced the incidence of fentanyl requirement by 30% and the number of fentanyl doses by 50% (P < 0.001). It also equalized the effects of naproxen, paracetamol and the placebo making the pain model invalid for this kind of study. The drawback associated with better analgesia was a doubling of the incidence of PONV (P < 0.001). CONCLUSIONS: Oral naproxen (10 mg/kg), but not oral paracetamol (20 mg/kg), reduces the need for rescue analgesic after adenoidectomy in children. The sensitivity of the pain model is crucial for these types of studies. PMID- 17465971 TI - Misplaced and dislodged endotracheal tubes may be detected by the defibrillator during cardiopulmonary resuscitation. AB - We present two cases of unrecognized endotracheal tube misplacements in out-of hospital cardiopulmonary resuscitation recognized by the analysis of transthoracic impedance. In Case 1, ventilation-induced changes in transthoracic impedance disappeared after an intubation attempt corresponding to oesophageal intubation. This was clinically recognized after several minutes, the endotracheal tube was repositioned and alterations in transthoracic impedance resumed. In Case 2, the initial ventilation-induced signal change following endotracheal intubation weakened after a few minutes. At that time, the defibrillator gave vocal and visual feedback to the rescuers on ventilatory inactivity, a pharyngeal air leak was discovered simultaneously and the tube was found to be dislodged. Continuous monitoring of transthoracic impedance provided by the defibrillator during cardiopulmonary resuscitation may contribute to the early detection of an initially misplaced or later dislodged endotracheal tube. PMID- 17465972 TI - Interprofessional education. PMID- 17465973 TI - Thoracic spinal anesthesia for gastrostomy in a patient with severe lung disease. PMID- 17465974 TI - Use of CobraPLUS for airway management in a child with tracheomalacia. PMID- 17465975 TI - Effect of age on pulmonary gas exchange during laparoscopy in the Trendelenburg lithotomy position. AB - BACKGROUND: Physiological changes in respiratory mechanics caused by aging may lead to a deterioration in pulmonary gas exchange, an increase in the alveolar arterial oxygen gradient [(A-a)D(O2)] and a difference between the arterial carbon dioxide (CO(2)) tension (P(a)(CO(2))) and expired end-tidal CO(2) tension (P(ET)(CO(2))) [P((a-ET))(CO(2))] during laparoscopy in the Trendelenburg lithotomy position (TLP). METHODS: The subjects were 51 gynecologic patients. Pressure-controlled ventilation was used to maintain P(ET)(CO(2)), measured by the side stream method, within the range 4-4.67 kPa. During laparoscopy with CO(2) insufflation in TLP, the tidal volume was increased to keep P(ET)(CO(2)) within +/- 20% of the pre-insufflation value. The subjects were divided into three groups by age: young group (< 45 years); middle-aged group (45-64 years); and elderly group ( > or = 65 years). RESULTS: Before pneumoperitoneum (PPN), significant differences were found between the young and elderly groups in the arterial oxygen tension (P(a)(O(2))), (A-a)D(O(2)), P(a)(CO(2)) and P((a ET))(CO(2)). In all groups, the peak inspiratory pressure and P(a)(CO(2)) increased progressively during PPN in TLP. P((a-ET))(CO(2)) increased gradually after starting CO(2) insufflation in TLP only in the elderly group. CONCLUSIONS: An increase in P((a-ET))(CO(2)) was seen during PPN in TLP in the elderly group. With CO(2) insufflation in TLP, the setting of mechanical ventilation based on the value of P(ET)(CO(2)) (measured by the side stream method) should be determined with caution in elderly patients. PMID- 17465976 TI - Accuracy in identifying patients with loss of consciousness in a police-operated emergency call centre - first step in the chain of survival. AB - BACKGROUND: The first link in the 'chain of survival' is the activation of Emergency Medical Services (EMS). In the major part of Denmark, police officers operate the alarm 1-1-2 centre, including calls for EMS. Our aim was to study the police 1-1-2 operators' accuracy in identifying calls concerning patients with loss of consciousness as a key symptom of life-threatening conditions. 'Unconsciousness' was defined as patients with a Glasgow Coma Scale (GCS) score of < 9, scored by the on-scene anaesthesiologist from the Mobile Emergency Care Unit (MECU). METHODS: This study was an observational cohort study based on data from the Police 1-1-2 Database and the Aarhus County Pre-hospital Database containing data from MECU cases during 6 months in 2004-05. RESULTS: Two thousand, three hundred and forty-three emergency calls with MECU dispatch were identified. In 1655 cases, both 1-1-2 data and the GCS score were recorded. Two hundred and ninety-five patients were found with a GCS score of < 9 at MECU arrival, 243 of whom were reported 'unconscious' by 1-1-2, giving a sensitivity of 82%. One thousand, three hundred and sixty patients were found with a GCS score of > or = 9, 972 of whom were reported 'awake', giving a specificity of 72%. The positive predictive value (percentage of patients found with a GCS score of < 9 at MECU arrival amongst patients reported as 'unconscious') was 39%. CONCLUSIONS: The accuracy was moderate with room for improvement. The positive predictive value was low, indicating over-triage of MECU. PMID- 17465977 TI - Propofol versus alfentanil to prevent movement responses during uterine curettage. AB - BACKGROUND: Propofol anaesthesia is frequently associated with movement responses in non-paralysed patients. Opioids decrease the probability of movement during noxious stimulation. Heart rate variability and frontal electromyography (EMG), which are related to subcortical functions, may be more closely related than surface electroencephalography (EEG) to movement responses to noxious stimulation. METHODS: Eighty-two patients scheduled for uterine dilatation and curettage were randomized to receive at the first intra-operative movement either a supplemental alfentanil bolus, 0.5 mg intravenously, or a supplemental propofol bolus, 0.7 mg/kg intravenously. The incidences of recurring movement during the procedure were compared between the two groups. The associations of a measure of heart rate variability (Anemon index), heart rate, EMG and two EEG variables with movement responses were evaluated. RESULTS: The incidences of recurring movement were 73% and 38% in the alfentanil and propofol groups, respectively [difference, 35%; 95% confidence interval, 9-56%; P= 0.014 between the groups). The Anemon index, heart rate, EMG and surface EEG variables displayed mainly reactive associations with movement responses. CONCLUSION: During uterine curettage under propofol-alfentanil-nitrous oxide anaesthesia, a propofol bolus of 0.7 mg/kg was more effective in preventing the recurrence of movement responses than an alfentanil bolus of 0.5 mg. Several physiological variables may be used to track significant arousal reactions, but not to predict them. PMID- 17465979 TI - A novel role of peroxin PEX6: suppression of aging defects in mitochondria. AB - Yeast cells become older with each division, but their daughters are born young. Mutational analysis shows that maintenance of this age asymmetry requires segregation of a complement of active mitochondria to daughters and that this process breaks down in older mother cells. This decline has implications for stem cell aging in higher organisms. PEX6, a peroxisome biogenesis gene, has been isolated as a multicopy suppressor of an atp2 age asymmetry mutant. Suppression depended on the presence of particular amino acid residues in Atp2p, and required adenosine triphosphate (ATP) binding and/or ATP hydrolysis activity of Pex6p. Extra copies of PEX6 corrected the deficit in Atp2p in mitochondria in the mutant by improving its import kinetics, resulting in near normal mitochondrial inheritance by daughter cells. The novel function of Pex6p described here may provide insights into peroxisomal and mitochondrial disorders and into metabolic diseases in general. PMID- 17465978 TI - Expansion of the calcium hypothesis of brain aging and Alzheimer's disease: minding the store. AB - Evidence accumulated over more than two decades has implicated Ca2+ dysregulation in brain aging and Alzheimer's disease (AD), giving rise to the Ca2+ hypothesis of brain aging and dementia. Electrophysiological, imaging, and behavioral studies in hippocampal or cortical neurons of rodents and rabbits have revealed aging-related increases in the slow afterhyperpolarization, Ca2+ spikes and currents, Ca2+transients, and L-type voltage-gated Ca2+ channel (L-VGCC) activity. Several of these changes have been associated with age-related deficits in learning or memory. Consequently, one version of the Ca2+ hypothesis has been that increased L-VGCC activity drives many of the other Ca2+-related biomarkers of hippocampal aging. In addition, other studies have reported aging- or AD model related alterations in Ca2+ release from ryanodine receptors (RyR) on intracellular stores. The Ca2+-sensitive RyR channels amplify plasmalemmal Ca2+ influx by the mechanism of Ca2+-induced Ca2+ release (CICR). Considerable evidence indicates that a preferred functional link is present between L-VGCCs and RyRs which operate in series in heart and some brain cells. Here, we review studies implicating RyRs in altered Ca+ regulation in cell toxicity, aging, and AD. A recent study from our laboratory showed that increased CICR plays a necessary role in the emergence of Ca2+-related biomarkers of aging. Consequently, we propose an expanded L-VGCC/Ca2+ hypothesis, in which aging/pathological changes occur in both L-type Ca2+ channels and RyRs, and interact to abnormally amplify Ca2+ transients. In turn, the increased transients result in dysregulation of multiple Ca2+-dependent processes and, through somewhat different pathways, in accelerated functional decline during aging and AD. PMID- 17465980 TI - Dynamics of the action of dFOXO on adult mortality in Drosophila. AB - The insulin/insulin growth factor (IGF)-like signaling (IIS) pathway has a conserved role in regulating lifespan in Caenorhabditis elegans, Drosophila and mice. Extension of lifespan by reduced IIS has been shown in C. elegans to require the key IIS target, forkhead box class O (FOXO) transcription factor, DAF 16. dFOXO, the Drosophila DAF-16 orthologue, is also an IIS target, and its overexpression in adult fat body increases lifespan. In C. elegans, IIS acts exclusively during adulthood to determine adult survival. We show here, using an inducible overexpression system, that in Drosophila continuous dFOXO overexpression in adult fat body reduces mortality rate throughout adulthood. We switched the IIS status of the flies at different adult ages and examined the effects of these switches on dFOXO expression and mortality rates. dFOXO protein levels were switched up or down by the inducible expression system at all ages examined. If IIS status is reversed early in adulthood, similar to the effects of another intervention that reduces adult mortality in Drosophila, dietary restriction (DR), there is a complete switch of subsequent mortality rate to that of flies chronically exposed to the new IIS regime. At this age, IIS thus acts acutely to determine risk of death. Mortality rates continued to respond to a switch in IIS status up to 4 weeks of adult age, but not thereafter. However, unlike DR, as IIS status was altered at progressively later ages, mortality rates showed incomplete switching and responded with progressively smaller changes. These findings indicate that alteration of expression levels of dFOXO may have declining effects on IIS status with age, that there could be some process that prevents or lessens the physiological response to a switch in IIS status or that, unlike DR, this pathway regulates aging-related damage. The decreased mortality and increased lifespan of dFOXO overexpressing flies was uncoupled from any effect on female fecundity and from expression levels of Drosophila insulin-like peptides in the brain. PMID- 17465981 TI - Changes in the structural complexity of the aged brain. AB - Structural changes of neurons in the brain during aging are complex and not well understood. Neurons have significant homeostatic control of essential brain functions, including synaptic excitability, gene expression, and metabolic regulation. Any deviations from the norm can have severe consequences as seen in aging and injury. In this review, we present some of the structural adaptations that neurons undergo throughout normal and pathological aging and discuss their effects on electrophysiological properties and cognition. During aging, it is evident that neurons undergo morphological changes such as a reduction in the complexity of dendrite arborization and dendritic length. Spine numbers are also decreased, and because spines are the major sites for excitatory synapses, changes in their numbers could reflect a change in synaptic densities. This idea has been supported by studies that demonstrate a decrease in the overall frequency of spontaneous glutamate receptor-mediated excitatory responses, as well as a decrease in the levels of alpha-amino-3-hydroxy-5-methylisoxazole-4 propionic acid and N-methyl-d-aspartate receptor expression. Other properties such as gamma-aminobutyric acid A receptor-mediated inhibitory responses and action potential firing rates are both significantly increased with age. These findings suggest that age-related neuronal dysfunction, which must underlie observed decline in cognitive function, probably involves a host of other subtle changes within the cortex that could include alterations in receptors, loss of dendrites, and spines and myelin dystrophy, as well as the alterations in synaptic transmission. Together these multiple alterations in the brain may constitute the substrate for age-related loss of cognitive function. PMID- 17465982 TI - No evidence for a clear link between active intestinal inflammation and autism based on analyses of faecal calprotectin and rectal nitric oxide. AB - AIM: Due to parental concern regarding the child's bowel habits and the ongoing discussion whether there might be an association between autism and intestinal inflammation, two inflammatory markers were analysed in a group of children with autism. METHODS: Twenty-four consecutive children with autism (3-13 years) of unknown aetiology were investigated with respect to faecal calprotectin and rectal nitric oxide (NO). RESULTS: One child who previously had a severe Clostridium difficile infection displayed raised levels of both these inflammatory markers and one child with extreme constipation for whom only calprotectin was possible to measure had raised levels. The remaining children displayed results that did not indicate an active inflammatory status in the intestine when the two inflammatory markers were combined. CONCLUSION: By the use of two independent markers of inflammatory reactions in the gut, i.e. rectal NO and faecal calprotectin we were not able to disclose evidence of a link between the autistic disorder and active intestinal inflammation. PMID- 17465983 TI - Shed some light on darkness: will Tanzania reach the millennium development goals? AB - The overall picture of health in sub-Saharan Africa can easily be painted in dark colours. The aim of this viewpoint is to discuss epidemiological data from Tanzania on overall health indicators and the burden of malaria and HIV. Is the situation in Tanzania improving or deteriorating? Are the health-related millennium development goals (MDG) on reducing under-five mortality, reducing maternal mortality and halting HIV and malaria within reach? CONCLUSION: Child mortality and infant mortality rates are decreasing quite dramatically. Malaria prevention strategies and new effective treatment are being launched. The MDG 4 on child mortality is clearly within reach, and the same optimism may apply to MDG 6 on combating malaria. PMID- 17465984 TI - Varicella seroprevalence in Turkish population in Cyprus. AB - AIM: This study was conducted to determine the age-specific seroprevalence of varicella zoster virus (VZV) infection in Turkish population in Cyprus. METHODS: A total of 600 unvaccinated individuals aged 1-30 years were selected for the study with cluster sampling. Information on socio-demographic characteristics was gathered for each participant and, anti-VZV antibodies were assayed by using enzyme immune assay. RESULTS: Of the 578 assayed samples, 486 (84.1%) were seropositive. Varicella seroprevalence increased sharply with age from 25% for the 2-3 year olds to 55, 78 and 85% for 4-5, 6-7 and 8-9 year olds, respectively. More than 90% of individuals >16 years of age were seropositive. Varicella seroprevalence was higher in large families with five and more members (91.2%) than in small families with four or fewer members (80.2%). CONCLUSION: The majority of varicella-zoster virus infections occur during preschool period and at the first years of schooling. Therefore, routine varicella vaccination of children would be logical in Northern Cyprus, as is currently recommended by the European Working Group on Varicella. PMID- 17465985 TI - Time of perinatal immunization, thimerosal exposure and neurodevelopment at 6 months in breastfed infants. AB - AIM: Brazilian newborns immunized with hepatitis-B (thimerosal containing vaccine, TCV) receive the first dose within 24 h if delivered in public hospitals, but at a later time if born in private hospitals. We compared neurodevelopment (ND) in infants born in a state hospital (immunized within 24 h) and in privately run hospitals (immunized 2-4 days postnatally). METHODS: We used the Gesell Developmental Schedules in 82 healthy exclusively breastfed infants at 6 months to assess motor skills, language development, comprehension capacity and social skills. RESULTS: Compared to the group immunized 2-4 days after hospital discharge, the group immunized within 24 h showed no significant difference in ND delays. Despite the variation in gestational age (range 36-42 weeks) and TCV ethylmercury (EtHg) dose (5.7-11.3 microg Hg/kg b.w.) at birth, time of exposure to TCV showed no significant association with ND. Gesell Developmental Score was not significantly correlated with total parenteral EtHg/unit of body mass neither with the relative increase in hair-Hg (as an additional challenge to prenatal Hg exposure). CONCLUSION: In breastfed infants, differences in early exposure to TCV EtHg cannot portend clinical neurodevelopment delays at 6 months. We speculate that breastfeeding remains a significant strategy to improve central nervous system protection of infants facing early exposure challenges. PMID- 17465986 TI - Inflammation at birth and the insulin-like growth factor system in very preterm infants. AB - BACKGROUND: Foetal inflammation is associated with an increased risk of brain damage in preterm infants whereas IGF-I is essential for cerebral development and exhibits anti-apoptotic properties. AIM: To assess levels of IGF-I and IGF binding proteins at very preterm birth and to evaluate their relationship with foetal pro-inflammation and cerebral damage. METHODS: Levels of IGF-I, IGF binding protein 3 (IGFBP-3), high- (hp) and low-phosphorylated (lp) IGFBP-1 in cord blood and neonatal blood at 72 h after delivery were analysed in relation to levels of cytokines and cerebral damage as detected by ultrasound in 74 inborn infants [mean gestational age (GA) 27.1 weeks]. Evaluation was performed separately according to birth weight for GA. RESULTS: In cord blood of infants appropriate for gestational age (AGA) higher levels of IL-6 and IL-8 were associated with lower IGF-I (r =-0.38, p = 0.008 and r =-0.36, p = 0.014). Higher levels of IL-6, IL-8 and TNF-alpha were associated with both higher levels of lpIGFBP-1 (r = 0.54, p < 0.001, r = 0.50, p < 0.001 and r = 0.13, p = 0.012, respectively) and hpIGFBP-1 (r = 0.55, p < 0.001, r = 0.45, p = 0.002 and r = 0.32, p = 0.026, respectively). Infants with intraventricular haemorrhage grade III (n = 5) had higher levels of lp/hpIGFBP-1 in cord blood (p = 0.001 and 0.002, respectively). CONCLUSION: Pro-inflammation at birth is associated with changes in the IGF-system. This may be of importance for development of brain damage in preterm infants. PMID- 17465987 TI - Indices of iron deficiency and anaemia in Bedouin and Jewish toddlers in southern Israel. AB - AIMS: To estimate the prevalence of iron deficiency and iron deficiency anaemia using haematological indices. METHODS: Prospective interventional study. Healthy toddlers from Bedouin and Jewish towns in southern Israel. Capillary blood was sampled to assess iron status and nutritional history recorded. Parents were given specific nutritional instructions. Anaemia was defined as haemoglobin level < or = 11 gr/dL. Iron deficiency without anaemia was defined as mean corpuscular volume (MCV) < 70 fL and/or red blood cell width (RDW) > or = 16, with haemoglobin level > 11 gr/dL. Toddlers with iron deficiency were treated with 5 mg/kg/day of elemental iron. Follow-up iron and nutritional status was performed 1 and 2 months after enrolment. RESULTS: At the time of enrolment 42% of the 107 Jewish and 93% of the 43 Bedouin toddlers were iron deficient (p < 0.001). Significantly higher proportions of Bedouin toddlers were breastfed, drank tea, did not eat meat, did not take supplementary iron in their first year of life and were previously diagnosed with anaemia. Rate of follow-up was 55% among Bedouins and 33% among Jews. The mean haemoglobin rise over two months was 0.91 gr/dL (95% CI: 0.63-1.18 gr/dL; p < 0.001) in Bedouins and 0.58 gr/dL (95% CI: 0.14-1.02 gr/dL; p = 0.014) in Jews. CONCLUSIONS: Higher rates of anaemia and iron deficiency, as well as most of the risk factors, found among the Bedouin toddlers, call for the design and implementation of innovative, culturally appropriate interventions in the Bedouin population. PMID- 17465988 TI - Effects of formalin fixation, paraffin embedding, and time of storage on DNA preservation in brain tissue: a BrainNet Europe study. AB - There is a large amount of tissue stored in brain collections and brain banks, but little is known about whether formalin-fixed tissues and paraffin blocks stored for years in brain banks are suitable for the retrospective genetic studies. The study was carried out in order to: (i) compare DNA preservation in frozen, formalin-fixed and paraffin-embedded tissues stored for different periods; (ii) study point mutations and triplet expansions in frozen, formalin fixed and paraffin-embedded material stored for variable periods, and using different fixative solutions; (iii) compare different methods to optimize DNA extraction and DNA amplification from suboptimally preserved brain tissue. DNA preservation is suitable for genetic studies in samples stored at -80 degrees C for several years. Formalin-fixed, paraffin-embedded tissue was inferior to frozen tissue, but did yield adequate results in many cases depending on the type of fixative solution and time of fixation before embedding. Prolonged fixation in formalin rarely yielded useful DNA. Similar results were obtained in samples from prion diseases. The best results were obtained by using the Qiagen kits (QIAmp DNA Micro) in frozen material, paraffin blocks and formalin-fixed tissue. Genomiphi and TaKaRa Ex Taq methods were also assayed in paraffin blocks and in formalin-fixed samples with limited success. PMID- 17465989 TI - Genome wide copy number abnormalities in pediatric medulloblastomas as assessed by array comparative genome hybridization. AB - Array-based comparative genomic hybridization was used to characterize 22 medulloblastomas in order to precisely define genetic alterations in these malignant childhood brain tumors. The 17p(-)/17q(+) copy number abnormality (CNA), consistent with the formation of isochromosome 17q, was the most common event (8/22). Amplifications in this series included MYCL, MYCN and MYC previously implicated in medulloblastoma pathogenesis, as well as novel amplicons on chromosomes 2, 4, 11 and 12. Losses involving chromosomes 1, 2, 8, 10, 11, 16 and 19 and gains of chromosomes 4, 7, 8, 9 and 18 were seen in greater than 20% of tumors in this series. A homozygous deletion in 11p15 defines the minimal region of loss on this chromosome arm. In order to map the minimal regions involved in losses, gains and amplifications, we combined aCGH data from this series with that of two others obtained using the same RPCI BAC arrays. As a result of this combined analysis of 72 samples, we have defined specific regions on chromosomes 1, 8p, 10q, 11p and 16q which are frequently involved in CNAs in medulloblastomas. Using high density oligonucleotide expression arrays, candidate genes were identified within these consistently involved regions in a subset of the tumors. PMID- 17465990 TI - Congenital glioblastoma: a clinicopathologic and genetic analysis. AB - Congenital central nervous system (CNS) tumors are uncommon, accounting for 1% of all childhood brain tumors. They present clinically either at birth or within the first 3 months. Glioblastoma (GBM) only rarely occurs congenitally and has not been fully characterized. We examined clinicopathologic features and genetic alterations of six congenital GBMs. Tumors were seen by neuroimaging as large, complex cerebral hemispheric masses. All showed classic GBM histopathology, including diffuse infiltration, dense cellularity, GFAP-positivity, high mitotic activity, endothelial proliferation and pseudopalisading necrosis. Neurosurgical procedures and adjuvant therapies varied. Survivals ranged from 4 days to 7.5 years; two of the three long-term survivors received chemotherapy, whereas the three short-term survivors did not. Paraffin-embedded tissue sections were used for FISH analysis of EGFR, chromosomes 9p21 (p16/CDKN2A) and 10q ( PTEN/DMBT1); sequencing of PTEN and TP53; and immunohistochemistry for EGFR and p53. We uncovered 10q deletions in two cases. No EGFR amplifications, 9p21 deletions, or mutations of TP53 or PTEN were noted; however, nuclear p53 immunoreactivity was strong in 5/6 cases. Tumors were either minimally immunoreactive (n = 3) or negative (n = 3) for EGFR. We conclude that congenital GBMs show highly variable survivals. They are genetically distinct from their adult counterparts and show a low frequency of known genetic alterations. Nonetheless, the strong nuclear expression of p53 in these and other pediatric GBMs could indicate that p53 dysregulation is important to tumorigenesis. PMID- 17465991 TI - Cytoplasmic extracts from adipose tissue stromal cells alleviates secondary damage by modulating apoptosis and promotes functional recovery following spinal cord injury. AB - Spinal cord injury (SCI) typically results from sustained trauma to the spinal cord, resulting in loss of neurologic function at the level of the injury. However, activation of various physiological mechanisms secondary to the initial trauma including edema, inflammation, excito-toxicity, excessive cytokine release and apoptosis may exacerbate the injury and/or retard natural repair mechanisms. Herein, we demonstrate that cytoplasmic extracts prepared from adipose tissue stromal cells (ATSCs) inhibits H(2)O(2)-mediated apoptosis of cultured spinal cord-derived neural progenitor cells (NPCs) resulting in increased cell survival. The ATSC extracts mediated this effect by decreasing caspase-3 and c-Jun-NH2 terminal kinase (SAPK/JNK) activity, inhibiting cytochrome c release from mitochondria and reducing Bax expression levels in cells. Direct injection of ATSC extracts mixed with Matrigel into the spinal cord immediately after SCI also resulted in reduced apoptotic cell death, astrogliosis and hypo-myelination but did not reduce the extent of microglia infiltration. Moreover, animals injected with the ATSC extract showed significant functional improvement of hind limbs as measured by the BBB (Basso, Beattie and Bresnahan) scale. Collectively, these studies show a prominent therapeutic effect of ATSC cytoplasmic extracts on SCI principally caused by an inhibition of apoptosis-mediated cell death, which spares white matter, oligodendrocytes and neurons at the site of injury. The ability of ATSC extracts to prevent secondary pathological events and improve neurologic function after SCI suggests that extracts prepared from autologous cells harvested from SCI patients may have clinical utility. PMID- 17465992 TI - Strong suppression of tumor growth by insulin-like growth factor-binding protein related protein 1/tumor-derived cell adhesion factor/mac25. AB - Insulin-like growth factor binding protein-related protein 1 (IGFBP-rP1) has been shown to induce cellular senescence or apoptosis of breast and prostate cancer cell lines in vitro. To examine whether IGFBP-rP1 acts as a tumor-suppressive protein in vivo, we established two model systems. Expression of IGFBP-rP1 in the human bladder carcinoma cell line EJ-1 was blocked by RNA interference. Human colon cancer cell line DLD-1, which did not express endogenous IGFBP-rP1, was transfected with an IGFBP-rP1 expression vector. When injected intraperitoneally or subcutaneously into nude mice, the IGFBP-rP1-expressing EJ-1 and DLD-1 cell lines grew poorly, whereas the IGFBP-rP1 non-producers grew rapidly and produced large tumors. In monolayer culture the IGFBP-rP1 producers and non-producers grew similarly in each model, whereas in soft agar culture the former produced far less colonies than the latter. The IGFBP-rP1 producers had IGFBP-rP1 bound to the cell surface, and adhered more efficiently to fibronectin and laminin-5 than the respective non-producers. Expression of IGFBP-rP1 did not affect the efficiency of insulin signaling. These results demonstrate that IGFBP-rP1 strongly suppresses tumor growth by an insulin-independent or insulin-like growth factor independent mechanism. Cell surface IGFBP-rP1 may reduce the anchorage independent growth ability, leading to the marked loss of tumorigenicity. PMID- 17465993 TI - FOXP3 gene expression in a tuberculosis case contact study. AB - Regulatory T lymphocytes (T(regs)) that express FOXP3 are involved in the beneficial attenuation of immunopathology, but are also implicated in down regulation of protective responses to infection. Their role in tuberculosis (TB) is unknown. We classified 1272 healthy TB contacts according to their tuberculin skin test (TST) and interferon (IFN)-gamma enzyme-linked immunospot (ELISPOT) results and 128 TB cases, and studied the expression of FOXP3 and interleukin (IL)-10 in blood samples. Compared to the uninfected contact group (TST(-), ELISPOT(-)), we observed higher levels of FOXP3 mRNA in blood from TB patients (< 0.001), but IL-10 expression was slightly lower (P = 0.04). In contrast, FOXP3 expression levels were significantly lower (P = 0.001) in the recently infected contacts (TST(+), ELISPOT(+)) but there was no difference for IL-10 (P = 0.74). We hypothesize that during early/subclinical TB, most of which will become latent, FOXP3(+) T(regs) may be sequestered in the lungs, but when TB becomes progressive, FOXP3 reappears at increased levels in the periphery. While these findings do not reveal the role, beneficial or harmful, of T(regs) in TB, they emphasize the probable importance of these cells. PMID- 17465994 TI - Assessment of thyroid function and urinary and breast milk iodine concentrations in healthy newborns and their mothers in Tehran. AB - OBJECTIVE: To measure breast milk iodine (MI) and urinary iodine (UI) concentrations in healthy newborns and their nursing mothers from an iodine sufficient region to determine adequacy and to relate these parameters to thyroid function tests in mothers and infants. DESIGN: Cross-sectional. PATIENTS: Forty eight healthy neonates of 37 to 42 weeks' gestation with normal cord blood TSH values and their mothers were recruited in Tehran, Iran. MEASUREMENTS: Serum thyroid function tests were performed, and maternal and infant urinary iodine excretion, and maternal MI concentration were measured. RESULTS: Neonatal age was 12.9 +/- 3.9 (mean +/- SD; range 7-30) days and maternal age was 25.8 +/- 5 years. Median (range) UI in neonates was 271 microg/l (57-800) and in mothers was 107 microg/l (20-710). Median (range) MI was 148 microg/l (45-750). Neonatal and maternal UI did not correlate with serum thyroid function tests. UI < 150, 150 230, and > 230 microg/l was found in 20, 12.5, and 67.5% of neonates and 79.1, 14, and 7% of mothers, respectively. MI was < 150, 150-180, and > 180 microg/l in 52.4, 11.9, and 35.7% of mothers, respectively. CONCLUSIONS: Among euthyroid neonates, UI was adequate despite low median maternal UI and MI concentrations. There were no significant correlations between UI or MI and thyroid function tests in the mothers and infants. PMID- 17465995 TI - Focal uptake of (18)F-fluorodeoxyglucose by thyroid in patients with nonthyroidal head and neck cancers. AB - OBJECTIVE: To evaluate the prevalence and significance of focal thyroid lesions identified by (18)F-fluorodeoxyglucose (FDG)-positron emission tomography (PET) in patients with nonthyroidal head and neck cancers (HNC). PATIENTS AND MEASUREMENTS: Patients with histologically identified HNC who underwent FDG-PET and computed tomography (CT) were reviewed retrospectively. We evaluated the prevalence of patients with focal thyroid FDG uptake and the risk of malignancy and proper management in these patients. RESULTS: Of 689 HNC patients, 19 (2.8%) had focal thyroid FDG uptake. Of the 12 patients with a histological diagnosis by surgery or needle biopsy, 5 (41.7%) had carcinomas, 4 papillary and 1 follicular, whereas the others had benign thyroid lesions. The maximum standardized uptake value on PET was not sufficient to discriminate between malignant and benign thyroid lesions (8.4 +/- 13.2 vs. 4.2 +/- 4.0; P > 0.4). The identification of incidental thyroid diseases helped guide patient counselling and combined surgery with HNC and thyroidectomy. CONCLUSION: Focal thyroid lesions incidentally found on FDG-PET in patents with nonthyroidal HNC have a high probability of malignancy. These lesions deserve further diagnostic examination before HNC treatment to ensure adequate therapy for incidental thyroid cancers. PMID- 17465996 TI - Author's reply: the influences of hyperprolactinaemia and obesity on cardiovascular risk markers: effects of cabergoline therapy. PMID- 17465997 TI - A prospective, multicentre study to investigate the efficacy, safety and tolerability of octreotide LAR (long-acting repeatable octreotide) in the primary therapy of patients with acromegaly. AB - OBJECTIVE: To evaluate the efficacy, safety and tolerability of octreotide LAR (long-acting repeatable octreotide) in the primary therapy of acromegaly. DESIGN AND PATIENTS: Ninety-eight previously untreated acromegalics were recruited into this prospective multicentre study. A total of 68 patients successfully completed 48 weeks of the study period, received 12 doses of octreotide LAR 10-30 mg every 4 weeks, and constituted the population used for this analysis. MEASUREMENTS AND RESULTS: A clinically relevant reduction (i.e. to < or = 5 microg/l) in mean GH (mGH) was recorded in 72% of patients after 24 weeks of treatment, and 42% reached a 'safe' GH value (< or = 2.5 microg/l). At week 48, 16 more patients were considered partial GH responders (GH > 2.5 microg/l and < or = 5 microg/l) and 44% had reached a GH level < or = 2.5 microg/l. IGF-1 levels normalized in 38% and 34% of patients after 24 and 48 weeks of treatment, respectively. At study completion, 10 patients (14.7%) who had not normalized their IGF-1 levels had achieved at least a 50% decrement in this marker. In eight microadenoma patients, tumour volume decreased from a mean baseline level of 298 +/- 145 mm3 to 139 +/- 94 mm3 after 24 weeks and to 99 +/- 70 mm3 after 48 weeks of therapy. In 60 patients with macroadenoma, the corresponding values were 3885 +/- 5077 mm3 at baseline and 2723 +/- 3435 and 2406 +/- 3207 mm3 after 24 and 48 weeks, respectively. At weeks 24 and 48, a significant (> 20%) tumour volume reduction was reported in 63% and 75% of patients, respectively. A reduction in the severity of symptoms of acromegaly was observed early in treatment and was maintained throughout the study period. CONCLUSION: Octreotide LAR represents a viable alternative to surgery for primary treatment of acromegaly leading to a progressive regression of tumour volume, a sustained control of biochemical abnormalities and an adequate relief of symptoms of the disease. PMID- 17465998 TI - L-selectin gene polymorphisms in Graves' disease. AB - OBJECTIVE: Graves' disease (GD) is an autoimmune disorder with genetic predisposition. There is strong evidence that members of the selectin family participate in the interaction between leucocytes and the endothelium, as well as in inflammatory cell recruitment. Moreover, a high serum level of a soluble form of L-selectin (SL-selectin) has been reported in untreated GD patients. However, the impact of L-selectin polymorphisms on GD has not yet been investigated. The aim of the present study was to elucidate whether L-selectin gene polymorphisms were associated with the development of GD. SUBJECTS AND DESIGN: L-selectin gene polymorphisms were investigated in 230 Chinese GD patients and 208 healthy control subjects without antithyroid autoantibodies or a family history of autoimmune disorders. Two L-selectin SNPs were genotyped by the PCR-restriction fragment length polymorphism (PCR-RFLP) method. RESULTS: A C/T polymorphism at position -642 of the promoter region and a Pro213Ser (c.725 C-T) polymorphism in exon 6 were examined using PCR-RFLP. There was a significant increase in -642T allele frequency in GD patients compared with healthy controls (70 vs. 62%; P = 0.0126; P(c) = 0.0252). The frequency of the c.725C allele in exon 6 also appeared higher in GD patients than in controls. Haplotype analysis showed a significant decrease in the -642C/c.725T haplotype in GD patients (26 vs. 34%; P = 0.0095; Pc = 0.0190). However there was no association between polymorphisms and certain GD clinical phenotypes, including age of onset and ophthalmopathy. CONCLUSIONS: L-selectin gene polymorphisms are associated with GD susceptibility in Chinese patients. PMID- 17465999 TI - Serum leptin, oxidized low density lipoprotein and plasma asymmetric dimethylarginine levels and their relationship with dyslipidaemia in adolescent girls with polycystic ovary syndrome. AB - OBJECTIVE: The aim of this study was to investigate serum leptin, oxidized low density lipoprotein (ox-LDL) and asymmetric dimethylarginine (ADMA) levels and their interaction with dyslipidaemia in adolescents with polycystic ovary syndrome (PCOS). PATIENTS AND DESIGN: The study group consisted of 23 obese (obPCOS) and 21 nonobese girls with PCOS (nPCOS), and 31 lean healthy controls. PCOS was defined by the National Institutes of Health (NIH) criteria as the presence of chronic oligoanovulation and hyperandrogenism. Fasting leptin, ox LDL, ADMA and detailed lipid-lipoprotein profile were determined. Atherogenic index (AI) was calculated as [Total cholesterol - HDL cholesterol/HDL cholesterol]. Logarithmic transformations were made for ox-LDL. RESULTS: Total cholesterol, triglycerides, LDL cholesterol, very low density lipoprotein (VLDL) cholesterol, apolipoprotein B, lipoprotein A levels and AI were higher and apolipoprotein AI was lower in obPCOS compared to those in controls (P < 0.05). LDL cholesterol, apolipoprotein B and lipoprotein A levels were higher in nPCOS compared to controls (P < 0.05). ADMA and ox-LDL levels did not differ in the three groups. Leptin was significantly higher in obPCOS compared with that in the other two groups (P < 0.001) and it was correlated with triglycerides (r = 0.62), VLDL cholesterol (r = 0.45), lipoprotein A (r = 0.38) and AI (r = 0.43) in the PCOS group (P < 0.05). CONCLUSION: Our data demonstrate that ADMA and ox-LDL levels in adolescent PCOS subjects were not different than those in controls. Abnormal lipid profile was shown in obese and nonobese girls with PCOS and leptin was related with these lipid abnormalities in the PCOS subjects. PMID- 17466000 TI - Radioiodine treatment for benign thyroid diseases. AB - Radioiodine has been in use for over 60 years as a treatment for hyperthyroidism. Major changes in clinical practice have occurred with the realization that accurate dosimetry is incapable of avoiding the risks of hypothyroidism, while more accurate assessment of the risks of other adverse effects of radioiodine such as ophthalmopathy and carcinogenesis have become available. More is also known of the potential for pretreatment with an antithyroid drug to affect the outcome of radioiodine treatment. However, we are still uncertain of the benefits of radioiodine treatment in subclinical hyperthyroidism. During the last two decades there has been wider acceptance of radioiodine as a safe and effective therapy for benign, nontoxic goitre, coupled with waning enthusiasm for the use of levothyroxine, as the risks and benefits of this option have become more apparent. The use of recombinant TSH offers the prospect that radioiodine treatment of nontoxic goitre can be simplified and improved, although more studies of this strategy are urgently required. PMID- 17466001 TI - Novel polymorphisms and lack of mutations in the ACD gene in patients with ACTH resistance syndromes. AB - OBJECTIVE: ACTH resistance is a feature of several human syndromes with known genetic causes, including familial glucocorticoid deficiency (types 1 and 2) and triple A syndrome. However, many patients with ACTH resistance lack an identifiable genetic aetiology. The human homolog of the Acd gene, mutated in a mouse model of adrenal insufficiency, was sequenced in 25 patients with a clinical diagnosis of familial glucocorticoid deficiency or triple A syndrome. DESIGN: A 3.4 kilobase genomic fragment containing the entire ACD gene was analysed for mutations in all 25 patients. SETTING: Samples were obtained by three investigators from different institutions. PATIENTS: The primary cohort consisted of 25 unrelated patients, primarily of European or Middle Eastern descent, with a clinical diagnosis of either familial glucocorticoid deficiency (FGD) or triple A syndrome. Patients lacked mutations in other genes known to cause ACTH resistance, including AAAS for patients diagnosed with triple A syndrome and MC2R and MRAP for patients diagnosed with familial glucocorticoid deficiency. Thirty-five additional patients with adrenal disease phenotypes were added to form an expanded cohort of 60 patients. MEASUREMENTS: Identification of DNA sequence changes in the ACD gene in the primary cohort and analysis of putative ACD haplotypes in the expanded cohort. RESULTS: No disease-causing mutations were found, but several novel single nucleotide polymorphisms (SNPs) and two putative haplotypes were identified. The overall frequency of SNPs in ACD is low compared to other gene families. CONCLUSIONS: No mutations were identified in ACD in this collection of patients with ACTH resistance phenotypes. However, the newly identified SNPs in ACD should be more closely examined for possible links to disease. PMID- 17466002 TI - Relationship between serum hsCRP concentration and biochemical bone turnover markers in healthy pre- and postmenopausal women. AB - OBJECTIVE: Although osteoporosis and atherosclerosis seem to be related, the mechanisms are not yet understood. We previously observed that women with higher serum concentrations of high sensitivity C-reactive protein (hsCRP), a strong risk factor for atherosclerosis, had lower bone mineral density (BMD). However, the relationship of hsCRP level with bone turnover rate, an independent risk factor for osteoporotic fracture, is not known. DESIGN: Cross-sectional hospital based survey. PATIENTS: Apparently healthy pre- and postmenopausal women (n = 39 and 150, respectively). MEASUREMENTS: Urinary N-terminal telopeptide of type I collagen (NTx) and serum bone specific alkaline phosphatase (BALP) were measured using commercially available immunoassay kits. Serum hsCRP concentrations were measured by a particle-enhanced immunoturbidometric method. RESULTS: Both urinary NTx (gamma = 0.288, P < 0.001) and serum BALP (gamma = 0.260, P < 0.001) were positively correlated with serum hsCRP levels. Significance remained even after adjustment for age, body mass index and years since menopause (gamma = 0.257, P < 0.001, and gamma = 0.163, P = 0.027, respectively). Compared with subjects in the lowest hsCRP quartile (< or = 0.6 mg/l), those in the highest hsCRP quartile (> or = 1.6 mg/l) had significantly higher urinary NTx concentrations (P = 0.001) after adjustment for confounding variables. There was an increasing trend of serum BALP concentrations in the higher hsCRP quartile groups (P = 0.073). CONCLUSION: These findings suggest that low grade systemic inflammation may be a common linking factor between development of atherosclerosis and increased bone turnover rate. PMID- 17466003 TI - Perioperative plasma active and total ghrelin levels are reduced in acromegaly when compared with in nonfunctioning pituitary tumours even after normalization of serum GH. AB - OBJECTIVE: Ghrelin is a novel gastric peptide known to stimulate GH secretion, but the relationship between ghrelin and the GH-insulin-like growth factor (IGF) 1 axis in GH excess or deficiency is poorly understood. This study investigated dysregulation of ghrelin secretion in acromegaly and its short-term postoperative recovery. METHODS: A prospective study was conducted on eight patients who underwent complete transsphenoidal resection of GH-producing pituitary adenomas (acromegaly group) and 22 for endocrinologically nonfunctioning pituitary tumours (control group). Active and total plasma ghrelin levels were measured serially before and after surgery. RESULTS: Preoperative active and total plasma ghrelin concentrations (mean +/- SD; fmol/ml) were significantly reduced in acromegalic patients when compared with those in the controls (9.6 +/- 4.3 and 157.4 +/- 65.6 vs. 21.8 +/- 13.0 and 267.1 +/- 111.4; P = 0.023 and P = 0.021, respectively). Both levels were still significantly suppressed on postoperative Day 7 in the acromegaly group when compared with those in the control group (11.7 +/- 4.3 and 197.8 +/- 68.9 vs. 22.5 +/- 12.6 and 302.7 +/- 100.0; P = 0.038 and P = 0.018, respectively). The ratios of active to total ghrelin were not significantly different between the two groups before and after operation. In acromegalic patients, active and total ghrelin levels remained significantly suppressed even after normalization of serum GH levels. CONCLUSIONS: The putative negative feedback mechanism of GH on ghrelin secretion may in part account for the low ghrelin levels observed in acromegalic patients, and the mechanism may persist even after normalization of serum GH. PMID- 17466004 TI - Complex ABCC8 DNA variations in congenital hyperinsulinism: lessons from functional studies. AB - OBJECTIVE: Congenital hyperinsulinism (CHI) is a cause of persistent and severe hypoglycaemia in infancy. Mutations in the genes ABCC8 and KCNJ11 encoding SUR1 and Kir6.2, respectively, are the commonest cause of CHI. We investigated whether the possession of two DNA variants leading to coding changes in a single allele of ABCC8 can affect the potential mechanism of disease pathogenesis. DESIGN AND PATIENTS: We studied two patients with complex mutations in the ABCC8 gene with CHI and used in vitro studies to explore the potential disease mechanism and the contribution of the various mutant allelles. RESULTS: The first case had diffuse disease and was homozygous for the mutations D1193V and R1436Q in SUR1. Channel complexes containing the D1193V mutant were delivered to the plasma membrane and were functional and those containing R1436Q were also present at the plasma membrane but were nonfunctional. Combining the two mutations (SUR1D1193V/R1436Q) led to intracellular retention of the channel complex. In a second family, the patient had histologically focal disease and was heterozygous for two mutations from his father (G228D and D1471N) and one from his mother (V1572I). SUR1 G228D and D1471N singly or in combination led to intracellular retention of the channel complex and loss of function. By contrast, V1572I is trafficked appropriately and is functional, consistent with a mechanism of reduction to hemizygosity of paternal ABCC8 in focal disease. V1572I is likely to be a benign DNA variant. CONCLUSION: In one patient the combination of two coding variants led to intracellular retention of channel complex. In a second patient, functional studies allowed us to unravel the DNA variants likely to be causing the abrogation of ATP-sensitive K(+) channel function. PMID- 17466005 TI - Maternal serum adiponectin and infant birthweight: the role of adiponectin isoform distribution. AB - OBJECTIVE: Adiponectin is an insulin-sensitizing protein that circulates in oligomeric complexes, including trimers, hexamers and high-molecular-weight (HMW) multimers. In pregnant women, conflicting associations have been reported between maternal serum levels of total adiponectin (i.e. reflecting all isoforms) and infant birthweight. As the HMW complex has recently been proposed as the primary mediator of metabolic bioactivity, we hypothesized that differences in isoform distribution may underlie these conflicting reports. Therefore, we evaluated the relationship between maternal adiponectin isoforms and infant birthweight. DESIGN/PATIENTS/MEASUREMENTS: HMW and total adiponectin, as well as the ratio of HMW to total adiponectin (ratio known as S(A)), were measured in healthy pregnant Caucasian women (n = 58) undergoing an oral glucose tolerance test (OGTT), following an abnormal glucose challenge test. RESULTS: On univariate analysis adjusted for neonate gender and length of gestation, birthweight was positively correlated with weight gain in pregnancy (r = 0.29, P = 0.031) and inversely associated with the IS(OGTT) index of insulin sensitivity (r = -0.27, P = 0.041), total adiponectin (r = -0.31, P = 0.021), HMW adiponectin (r = -0.34, P = 0.0093) and S(A) (r = -0.34, P = 0.011). On multiple linear regression analyses, however, total adiponectin was not related to birthweight. By contrast, HMW adiponectin was related at borderline significance (t = -1.87, P = 0.068), while S(A) emerged as an independent negative determinant of infant birthweight (t = -2.46, P = 0.0175). Adjusted mean neonatal birthweight was significantly higher in the infants of women comprising the lowest tertile of S(A) compared to women in the highest tertile of S(A) (3684 vs. 3424 g, P = 0.0375). CONCLUSIONS: The proportion of maternal serum adiponectin in HMW form (S(A)) is independently and inversely associated with infant birthweight. Thus, adiponectin isoform distribution, rather than total adiponectin concentration, may be a correlate of foetal size. PMID- 17466006 TI - Efficacy and safety of orbital decompression in treatment of thyroid-associated ophthalmopathy: long-term follow-up of 78 patients. AB - OBJECTIVE: To study the long-term results of decompression surgery in patients with thyroid-associated ophthalmopathy (TAO). DESIGN AND METHODS: Retrospective analysis and clinical re-examination of TAO patients with comparison to healthy controls in Helsinki University Central Hospital. PATIENTS: Seventy-eight patients who had undergone orbital decompression due to TAO by transantral or endonasal technique between the years 1985 and 2000 were invited for re examination. For comparison, 79 healthy age- and sex-matched controls underwent a similar examination. RESULTS: In comparison to the patients' preoperative state, proptosis was reduced by 4.7 (2.6) mm (mean (SD)) in the right and 4.4 (2.9) mm in the left globe (P < 0.0001) but did not reach the level of the controls' globe (P < 0.0001). In the right and left eye of patients, visual acuity improved in 44 55%, remained stable in 27-36% and worsened in 18-20%, but remained significantly worse than in controls (P =0.02-0.001). Prevalence of diplopia was high at the re examination (N = 39, 50%), but 54% of those patients considered it less difficult than before the surgery. Among patients and controls, maxillary sinusitis and facial neuralgias were equally common. Sensory disturbances were significantly more frequent in patients (P < 0.0001). In assessment of overall satisfaction with present eye status, the median of the visual analogue scale (VAS) was lower for the patients than the controls (P < 0.0001). The majority of the patients considered the operation very helpful regardless of the technique used. CONCLUSION: For patients with TOA, orbital decompression seems to be an effective and safe treatment. PMID- 17466007 TI - The advantage of measuring spontaneous growth hormone (GH) secretion compared with the insulin tolerance test in the diagnosis of GH deficiency in young adults. AB - OBJECTIVE: Reassessment of GH status after the attainment of adult height has important clinical implications in the diagnosis and prognosis of GH deficiency (GHD) in adulthood. The current GH threshold for biochemical definition of GHD in young adults is still a subject of debate. DESIGN: To investigate the role of pharmacological stimulation tests compared with spontaneous 12-h nocturnal GH secretion in the diagnosis of permanent GHD in young adults with childhood-onset GHD. PATIENTS: Forty-five young adults (25 males, 20 females) with childhood onset GHD, height standard deviation score (SDS) -1.1 +/- 1.3 and body mass index (BMI) SDS 1.0 +/- 1.6, were re-evaluated at the age of 19.8 +/- 2.7 years. Sixteen subjects showed a normal pituitary gland on magnetic resonance imaging (MRI), while in 29, consistent structural hypothalamic-pituitary abnormalities were found. GH secretion was assessed by means of an insulin tolerance test (ITT) and a 12-h spontaneous nocturnal profile as well as by IGF-I assessment. The results were compared with those of 43 healthy controls. RESULTS: Mean 12-h spontaneous nocturnal GH secretion was < 3.1 microg/l (the lowest limit of the normal range) in 36 (80%) of the subjects and > 3.1 microg/l in nine (20%). Of these 36 patients, 29 (80%) had abnormal MRI findings and 20 (55%) had multiple pituitary hormone deficiencies (MPHD). All nine subjects with mean spontaneous GH secretion > 3.1 microg/l had a normal pituitary MRI, isolated GHD and a peak GH response to ITT > 5 microg/l. There was a discordance in 14 patients (31%), who showed a peak GH response to ITT > 5 microg/l but a reduced spontaneous GH secretory capacity; 10 had structural hypothalamic-pituitary abnormalities on MRI. CONCLUSIONS: Although the ITT provides valuable information and proves to be a sensitive index of permanent GHD, the results of this study emphasize the potential diagnostic value of assessment of 12-h spontaneous GH secretion in young adults with childhood-onset GHD. PMID- 17466008 TI - Can cortisol predict the future in obesity? PMID- 17466009 TI - Cortisol levels and measures of body composition in middle-aged and older men. AB - INTRODUCTION: Similarities in the symptomatic expressions of excess adiposity and hypercortisolaemic conditions suggest that elevated glucocorticoid exposure may influence the pathogenesis of obesity. Circulating cortisol levels are not typically elevated in obese subjects, but data from large prospective samples are rare. We undertook an analysis to determine both cross-sectional and longitudinal associations between body composition and serum cortisol concentrations in a randomly chosen group of 999 community-dwelling men, aged 40-79 years. METHODS: Data were obtained from the two follow-up waves of the Massachusetts Male Ageing Study (T2: 1995-97; T3: 2002-04). Partial correlation and multivariate regression analyses were used to estimate cross-sectional (T2) and longitudinal associations between serum cortisol concentrations and a range of measures of subjects' body composition, including weight, body mass index (BMI), waist circumference (WC), waist-to-hip girth ratio (WHR), and percentage body fat (measured by bioelectrical impedance at T3); similar analyses were conducted to assess the association between change (T2 to T3) in serum cortisol and simultaneous change in body composition parameters. RESULTS: We observed weak negative associations between cortisol concentrations and all body composition parameters, with the exception of percentage body fat. Longitudinal results demonstrated similar relationships but associations were of lesser magnitude. T2 cortisol concentrations were not associated with change in body composition over time, whereas T2 body size was positively associated with longitudinal changes in cortisol concentrations, providing limited evidence that weight change drives changes in cortisol concentrations, rather than vice versa. Results were unchanged when age and other covariate effects were controlled. CONCLUSIONS: Circulating cortisol concentrations are somewhat lower in obese than in nonobese community-dwelling men. There is some evidence that excess adiposity presages increases in cortisol concentrations, rather than the reverse. However, this observation should be greeted with caution, as age-related weight loss - and not gain - was associated with simultaneous increases in serum cortisol concentrations. PMID- 17466010 TI - Asymptomatic bilateral adrenal pheochromocytoma in a patient with a germline V804M mutation in the RET proto-oncogene. AB - A diagnosis of bilateral pheochromocytoma warrants exclusion of hereditary pheochromocytoma. OBJECTIVE: To describe the first case of a bilateral pheochromocytoma associated with V804M mutation in the RET proto-oncogene. PATIENTS: The index case was a 54-year-old man with bilateral adrenal masses discovered during a CT scan performed for other reasons. MEASUREMENTS: Genetic analysis included exons 8-11 and 13-17 in the RET proto-oncogene, all four exons and flanking intronic regions in the SDHD gene, all eight exons and flanking intronic regions in the SDHB, and all three exons in the VHL gene. RESULTS: Investigations revealed elevated urinary metanephrines (32.3 micromol/day), and laparoscopic bilateral adrenalectomy confirmed bilateral pheochromocytomas. A heterozygous V804M mutation in exon 14 of the RET was found in the index case and in four relatives. Total thyroidectomy, performed in four of five affected members in this kindred, disclosed a medullary thyroid carcinoma in the index case and in a 50-year-old woman, and nodular C-cell hyperplasia in the other two subjects. CONCLUSIONS: This clinical case suggests that individuals carrying the germline V804M mutation should be screened annually for the presence of pheochromocytoma. PMID- 17466011 TI - Phenotypic variability in 17beta-hydroxysteroid dehydrogenase-3 deficiency and diagnostic pitfalls. AB - OBJECTIVE: 17beta-hydroxysteroid dehydrogenase type 3 isoenzyme (17beta-HSD3) is required to produce testosterone for male sex differentiation. Mutations in the HSD17B3 gene cause 17betaHSD3 deficiency and result in XY sex reversal of varying degree. We report the phenotypes of 14 subjects with 17betaHSD3 deficiency in relation to sex of rearing, androgen production, and HSD17B3 mutations. DESIGN: Cases were identified through the Cambridge Disorders of Sex Development Database where detailed clinical information was recorded, results of hCG stimulation tests were available, and HSD17B3 mutation was identified. RESULTS: Fourteen subjects from seven pedigrees (four consanguineous) had the following seven mutations: A56T, N130S, E215D, S232L, C268Y, V205E, and a novel mutation M197K. XY sex reversal was classified as complete in 10 infants at birth. Inguinal masses suggestive of androgen insensitivity syndrome (AIS) occurred in five infants. Contrasexual virilization reminiscent of 5alpha-reductase deficiency occurred in four subjects at puberty. The median (range) testosterone : androstenedione (T/A) ratio after a short hCG stimulation test was 0.32 (0.12 3.4). The S232L mutation identified in three affected family members caused isolated, severe hypospadias in one member who was raised male; virilization occurred despite in vitro studies showing an inactive mutant enzyme. Ratios of T/A in this pedigree were more than 0.8. CONCLUSION: XY sex reversal is sufficiently variable in 17betaHSD3 deficiency to cause problems in accurate diagnosis, particularly in distinguishing it from AIS. It should be considered in undervirilized male infants with normal Wolffian duct structures, absent Mullerian ducts, and normal adrenal steroid biosynthesis; or when an assigned female subject virilizes at puberty. Elevated hCG-stimulated T/A ratio may occur, and sex of rearing may not be concordant within affected families with the same HSD17B3 mutation. The T/A ratio, mutation analysis and functional analysis of the mutant enzyme taken in isolation, respectively, may not conclusively establish a diagnosis of 17betaHSD3 deficiency in undervirilized male subjects; the reasons for these discrepancies remain unknown. PMID- 17466012 TI - Ethnic differences in TSH but not in free T4 concentrations or TPO antibodies during pregnancy. AB - OBJECTIVE: To describe the TSH, free T4 and thyroid peroxidase antibody (TPO-Ab) concentrations during pregnancy among four ethnic groups and to determine reference values for these parameters during normal pregnancy. METHODS: Cross sectional study of a cohort of 3270 pregnant women living in the city of Amsterdam. Blood samples were drawn at first booking in the first or second trimester. TSH, free T4 and TPO-Ab concentrations were determined. Four ethnic groups were studied: Dutch, Surinam, Turkish and Moroccan. RESULTS: Plasma TSH increased and free T4 decreased from the first to the second trimester of pregnancy for all the ethnic groups. Ethnic differences were observed in TSH concentrations, with Dutch females having the highest TSH levels compared to the other three ethnic groups. The median TSH difference was 0.16 mU/l between the Dutch and Moroccan women, 0.15 mU/l between the Dutch and Surinam women and 0.10 mU/l between the Dutch and Turkish women. These could not be explained by differences in age, parity and current smoking status. No differences were seen in free T4 concentrations between the four ethnic groups. The prevalence of TPO Ab was comparable across the ethnic groups (about 6% in each); the concentration of TPO-Ab was also comparable among the ethnic groups. The Dutch women had a higher lower-limit (2.5 percentile) of the TSH reference range than the Surinam, Turkish and Moroccan women, ranging from 0.14 mU/l for the Surinam and Moroccan to 0.27 mU/l for the Dutch women. CONCLUSION: The increase in TSH and decrease in free T4 values during pregnancy correspond to previous reported studies. Pregnant Dutch women had consistently higher TSH values than the ethnic group, but corresponding free T4 levels and TPO-Ab did not differ between these ethnic groups. PMID- 17466013 TI - Protein secretion systems in Mycobacteria. AB - Mycobacteria have a unique cell-envelope structure which protects the bacteria from the extracellular environment by limiting access to noxious molecules from the outside. This extremely hydrophobic and thick barrier also poses a unique problem for the export of bacterial products. Here we review the multiple protein secretion pathways in Mycobacteria, including the general secretion pathway and the Twin-Arginine Transporter, with an emphasis on the ESX-1 alternate secretion system. This newly identified protein secretion system is required for growth during infection and has provided insight into how M. tuberculosis manipulates the host immune response during infection. PMID- 17466014 TI - The co-ordinated regulation of iron homeostasis in murine macrophages limits the availability of iron for intracellular Salmonella typhimurium. AB - In being both, a modifier of cellular immune effector pathways and an essential nutrient for microbes, iron is a critical determinant in host-pathogen interaction. Here, we investigated the metabolic changes of macrophage iron homeostasis and immune function following the infection of RAW264.7 murine macrophages with Salmonella typhimurium. We observed an enhanced expression of the principal iron export protein, ferroportin 1, and a subsequent increase of iron efflux in Salmonella-infected phagocytes. In parallel, the expression of haem oxygenase 1 and of the siderophore-binding peptide lipocalin 2 was markedly enhanced following pathogen entry. Collectively, these modulations reduced both the cytoplasmatic labile iron and the ferritin storage iron pool within macrophages, thus restricting the acquisition of iron by intramacrophage Salmonella. Correspondingly, limitation of macrophage iron decreased microbial survival, whereas iron supplementation impaired immune response pathways in Salmonella-infected macrophages (nitric oxide formation and tumour necrosis factor-alpha production) and promoted intracellular bacterial proliferation. Our findings suggest that the enhancement of ferroportin 1-mediated iron efflux, the upregulation of the haem-degrading enzyme haem oxygenase 1 and the induction of lipocalin 2 following infection concordantly aim at withholding iron from intracellular S. typhimurium and to increase antimicrobial immune effector pathways thus limiting pathogen proliferation. PMID- 17466015 TI - Response to Olivarius and Siersma: weight change or change in weight trend. Differences in analytical approaches (and differences in study samples). PMID- 17466016 TI - Biochemical characterization of the minimal polyketide synthase domains in the lovastatin nonaketide synthase LovB. AB - The biosynthesis of lovastatin in Aspergillus terreus requires two megasynthases. The lovastatin nonaketide synthase, LovB, synthesizes the intermediate dihydromonacolin L using nine malonyl-coenzyme A molecules, and is a reducing, iterative type I polyketide synthase. The iterative type I polyketide synthase is mechanistically different from bacterial type I polyketide synthases and animal fatty acid synthases. We have cloned the minimal polyketide synthase domains of LovB as standalone proteins and assayed their activities and substrate specificities. The didomain proteins ketosynthase-malonyl-coenzyme A:acyl carrier protein acyltransferase (KS-MAT) and acyl carrier protein-condensation (ACP-CON) domain were expressed solubly in Escherichia coli. The monodomains MAT, ACP and CON were also obtained as soluble proteins. The MAT domain can be readily labeled by [1,2-(14)C]malonyl-coenzyme A and can transfer the acyl group to both the cognate LovB ACP and heterologous ACPs from bacterial type I and type II polyketide synthases. Using the LovB ACP-CON didomain as an acyl acceptor, LovB MAT transferred malonyl and acetyl groups with k(cat)/K(m) values of 0.62 min( 1).mum(-1) and 0.032 min(-1).mum(-1), respectively. The LovB MAT domain was able to substitute the Streptomyces coelicolor FabD in supporting product turnover in a bacterial type II minimal polyketide synthase assay. The activity of the KS domain was assayed independently using a KS-MAT (S656A) mutant in which the MAT domain was inactivated. The KS domain displayed no activity towards acetyl groups, but was able to recognize malonyl groups in the absence of cerulenin. The relevance of these finding to the priming mechanism of fungal polyketide synthase is discussed. PMID- 17466017 TI - Characterization and structural modeling of a new type of thermostable esterase from Thermotoga maritima. AB - A bioinformatic screening of the genome of the hyperthermophilic bacterium Thermotoga maritima for ester-hydrolyzing enzymes revealed a protein with typical esterase motifs, though annotated as a hypothetical protein. To confirm its putative esterase function the gene (estD) was cloned, functionally expressed in Escherichia coli and purified to homogeneity. Recombinant EstD was found to exhibit significant esterase activity with a preference for short acyl chain esters (C4-C8). The monomeric enzyme has a molecular mass of 44.5 kDa and optimal activity around 95 degrees C and at pH 7. Its thermostability is relatively high with a half-life of 1 h at 100 degrees C, but less stable compared to some other hyperthermophilic esterases. A structural model was constructed with the carboxylesterase Est30 from Geobacillus stearothermophilus as a template. The model covered most of the C-terminal part of EstD. The structure showed an alpha/beta-hydrolase fold and indicated the presence of a typical catalytic triad consisting of a serine, aspartate and histidine, which was verified by site directed mutagenesis and inhibition studies. Phylogenetic analysis showed that EstD is only distantly related to other esterases. A comparison of the active site pentapeptide motifs revealed that EstD should be grouped into a new family of esterases (Family 10). EstD is the first characterized member of this family. PMID- 17466018 TI - Gene expression waves. Cell cycle independent collective dynamics in cultured cells. AB - The ergodic hypothesis, which assumes the independence of each cell of the ensemble from all the others, is a necessary prerequisite to attach single cell based explanations to the grand averages taken from population data. This was the prevailing view about the interpretation of cellular biology experiments that typically are performed on colonies of billions of cells. By analysing gene expression data of different cells going from yeast to mammalian cell cultures, we demonstrate that cell cultures display a sort of "ecology-in-a-plate" giving rise to a rich dynamics of gene expression that are independent from reproductive cycles, hence contradicting simple ergodic assumptions The aspecific character of the observed coordinated gene expression activity inhibits any simple mechanistic hypothesis and highlights the need to consider population effects in the interpretation of data coming from cell cultures. PMID- 17466019 TI - Proliferating neuronal progenitors in the postnatal hippocampus transiently express the proneural gene Ngn2. AB - Little is known of the transcription factors expressed by adult neural progenitors produced in the hippocampal neurogenic niche. Here, we study the expression of the proneural basic helix-loop-helix (bHLH) transcription factor Neurogenin-2 (Ngn2) in the adult hippocampus. We have characterized the pattern of expression of Ngn2 in the adult hippocampus using immunostaining for Ngn2 protein and a Ngn2-green fluorescent protein (GFP) reporter mouse strain. A significant proportion of Ngn2-expressing cells were mitotically active. Ngn2-GFP expression was restricted to the subgranular zone and declined with age. Neuronal markers were used to determine the phenotype of Ngn2-expressing cells. The vast majority of Ngn2-GFP-positive cells expressed the immature neuronal markers, doublecortin (DCX) and polysialic acid-neural cell adhesion molecule (PSA-NCAM). Finally, the pattern of Ngn2 expression was studied following seizure induction. Our data show an increase in neurogenesis, detected in these animals by bromodeoxyuridine (BrdU) and DCX staining that was contemporaneous with a marked increase in Ngn2-GFP-expression. Taken together, our results show that Ngn2-GFP represents a specific marker for neurogenesis and its modulation in the adult hippocampus. Ngn2 transient expression in proliferating neuronal progenitors supports the idea that it plays a significant role in adult neurogenesis. PMID- 17466020 TI - Regional and phenotype heterogeneity of cellular prion proteins in the human brain. AB - Transmissible spongiform encephalopathies (TSEs) are neurological disorders that include genetic, infectious and sporadic forms of human Creutzfeldt-Jakob disease (CJD). The pathogenic agent is the prion protein that is composed of an abnormal isoform (PrP(Sc)) of a host-encoded protein (PrP(C)). Analysis of the relative amounts of PrP(Sc) glycoforms has been used to discriminate between various agents involved in TSE. The distribution and efficiency of conversion to PrP(Sc) can be influenced by differences in the expression of PrP(C). However, little attention has been given so far to the banding patterns of PrP(C). Using four different antibodies recognizing amino- and carboxyl-terminal PrP sequences we analysed the glycoforms of PrP(C) in seven regions of the human brain using brains obtained from six subjects. For determination of the staining intensities, signals were quantified by densitometry and reproducible patterns were accomplished by many repeated immunoblot analyses. When amino-terminal binding antibodies were used for detection, PrP(C) in the frontal neocortex, nucleus lentiformis, thalamus, hippocampus and cerebellum displayed a glycotype with high staining of the diglycosylated isoforms. This was different from patterns in the pons and medulla oblongata, which showed a high intensity of the nonglycosylated isoform, and PrP(C) proteins, approximately 27 kDa in size, exhibited high staining using the carboxyl-terminal binding antibodies. This intense staining followed from an overlay of full-length and truncated PrP(C) isoforms. Furthermore, we found marked differences in the expression of PrP(C). Variations in the processing of PrP(C) may lead to interregional differences in the glycoform composition of PrP(Sc) in human brains. PMID- 17466021 TI - Nicotine gates long-term potentiation in the hippocampal CA1 region via the activation of alpha2* nicotinic ACh receptors. AB - Hippocampal CA1 pyramidal cells receive two major excitatory synaptic inputs via the Schaffer collateral (SC) and temporoammonic (TA) pathways. Nicotine promotes induction of long-term potentiation (LTP) in the SC path; however, it is not known whether the modulatory effect of nicotine on LTP induction is pathway specific. Here we show that nicotine suppresses LTP induction in the TA path. Interestingly, these opposing effects of nicotine were absent or greatly reduced in alpha2 nicotinic acetylcholine receptor (nAChR)-knockout (KO) mice, suggesting that an alpha2-containing nAChR subtype mediates these effects. Optical imaging with a voltage-sensitive dye revealed significantly stronger membrane depolarization in the presence of nicotine in the SC path, facilitating spread of excitatory neural activity along both the somatodendritic and the CA1 proximodistal axes. These effects of nicotine were also absent in alpha2 nAChR-KO mice, suggesting that the enhanced optical signal is related to the nicotine induced facilitation of LTP induction. In contrast, in the TA path nicotine terminated depolarization more quickly and increased the delayed hyperpolarization in the termination zone of the TA path input. These inhibitory effects of nicotine were absent in alpha2 nAChR-KO mice and, thus, most probably underlie the nicotine-induced suppression of LTP induction. Our results suggest that nicotine influences the local balance between excitation and inhibition, gates LTP, and directs information flow through the hippocampal circuits via the activation of alpha2* nAChRs. These effects of nicotine may represent the cellular basis of nicotine-mediated cognitive enhancement. PMID- 17466022 TI - Reversible inactivation of amygdala and cerebellum but not perirhinal cortex impairs reactivated fear memories. AB - The cerebellum, amygdala and perirhinal cortex are involved in fear learning but the different roles that these three structures play in aversive learning are not well defined. Here we show that in adult rats amygdala or cerebellar vermis blockade causes amnesia when performed immediately, but not 1 h, after the recall of fear memories. Thus, the cerebellum, as well as the amygdala, influences long term fear memories. These effects are long lasting, as they do not recover over time, even after a reminder shock administration. However, all of the subjects were able to form new fear memories in the absence of inactivation. By increasing the strength of conditioning, we observed that stronger fear memories are affected by the combined but not independent amygdala and cerebellar blockade. These results demonstrate that the cerebellum supports the memory processes even in the absence of a crucial site for emotions like the amygdala. Furthermore, they suggest that the amygdala is only one of the neural sites underlying long term fear memories. Finally, the inactivation of the perirhinal cortex never alters retrieved fear traces, showing important differences between the amygdala, cerebellum and perirhinal cortex in emotional memories. PMID- 17466023 TI - Ecophysiology of mycolic acid-containing Actinobacteria (Mycolata) in activated sludge foams. AB - Increasing incidences of activated sludge foaming have been reported in the last decade in Danish plants treating both municipal and industrial wastewaters. In most cases, foaming is caused by the presence of Actinobacteria; branched mycolic acid-containing filaments (the Mycolata) and the unbranched Candidatus'Microthix parvicella'. Surveys from wastewater treatment plants revealed that the Mycolata were the dominant filamentous bacteria in the foam. Gordonia amarae-like organisms and those with the morphology of Skermania piniformis were frequently observed, and they often coexisted. Their identity was confirmed by FISH, using a new permeabilization procedure. It was not possible to identify all abundant Mycolata using existing FISH probes, which suggests the presence of currently undetectable and potentially undescribed populations. Furthermore, some Mycolata failed to give any FISH signal, although substrate uptake experiments with microautoradiography revealed that they were physiologically active. Ecophysiological studies were performed on the Mycolata identified by their morphology or FISH in both foams and mixed liquors. Large differences were seen among the Mycolata in levels of substrate assimilation and substrate uptake abilities in the presence of different electron acceptors. These differences were ascribed mainly to the presence of currently undescribed Mycolata species and/or differences in foam age. PMID- 17466024 TI - Diversity of laccase-like multicopper oxidase genes in Morchellaceae: identification of genes potentially involved in extracellular activities related to plant litter decay. AB - Despite the important role played by soil-inhabiting ascomycetes in plant litter decay processes, studies on the diversity and function of their laccase-like multicopper oxidase (LMCO) genes are scarce. In the present work, the LMCO gene diversity in 15 strains representing nine Morchellaceae and one Discinaceae species was evaluated by PCR. One to six different genes were found within the species, representing 26 different sequence types. Cluster analysis revealed LMCO genes belonging to four main gene families encoding different protein classes (Class I-IV). To identify the genes related to extracellular activities and potentially involved in litter decay processes, liquid cultures were induced by different aromatic compounds. Morchella conica and Verpa conica showed the strongest LMCO activity enhancement in the presence of the naturally occurring phenolic compound guaiacol, and their expressed LMCO genes were identified by sequencing. Only genes belonging to the gene families encoding the Class II and III proteins were expressed. Both genes (Class II and III) of the mycorrhizal like strain M. conica were exclusively expressed in the presence of guaiacol. In contrast to the saprotrophic strain V. conica, the gene encoding the Class III protein was constitutively expressed as it was also found in control cultures without guaiacol. PMID- 17466025 TI - Strains of the cyanobacterial genera Calothrix and Rivularia isolated from the Baltic Sea display cryptic diversity and are distantly related to Gloeotrichia and Tolypothrix. AB - Strains of the cyanobacterial genus Calothrix display pronounced tapering filaments. These cyanobacteria are benthic, have a worldwide distribution and are among the most easily recognizable cyanobacterial genera. However, it is not clear whether cyanobacterial strains assigned to the genus Calothrix constitute a natural monophyletic group. We sequenced 16S rRNA genes from 42 cyanobacterial cultures and environmental samples belonging to the genus Calothrix, and the morphologically similar genera Rivularia, Gloeotrichia and Tolypothrix. Phylogenetic analysis of the 16S rRNA gene identified large sequence diversity among the Calothrix morphotype strains. Our results demonstrate that Calothrix, Gloeotrichia and Tolypothrix do not form a monophyletic group but instead display a high level of genetic diversity. The evolutionary distances between cyanobacteria, morphologically identified as Calothrix, suggest that they belong to at least five different genera. Our results also suggest that the genus Gloeotrichia is distantly related to the genus Calothrix. We found correlations between genetic grouping and morphology in redundancy analysis. However, morphology alone was not sufficiently reliable to distinguish strains from different 16S rRNA gene clusters. The high level of diversity that we observed confirms the hypothesis that the Rivulariaceae are species rich. PMID- 17466026 TI - Pigment composition and adaptation in free-living and symbiotic strains of Acaryochloris marina. AB - Acaryochloris marina strains have been isolated from several varied locations and habitats worldwide demonstrating a diverse and dynamic ecology. In this study, the whole cell photophysiologies of strain MBIC11017, originally isolated from a colonial ascidian, and the free-living epilithic strain CCMEE5410 are analyzed by absorbance and fluorescence spectroscopy, laser scanning confocal microscopy, sodium dodecyl sulfate polyacrylamide gel electrophoresis and subsequent protein analysis. We demonstrate pigment adaptation in MBIC11017 and CCMEE5410 under different light regimes. We show that the higher the incident growth light intensity for both strains, the greater the decrease in their chlorophyll d content. However, the strain MBIC11017 loses its phycobiliproteins relative to its chlorophyll d content when grown at light intensities of 40 microE m(-2) s( 1) without shaking and 100 microE m(-2) s(-1) with shaking. We also conclude that phycobiliproteins are absent in the free-living strain CCMEE5410. PMID- 17466027 TI - Environmental effects on grass-endophyte associations in the harsh conditions of south Patagonia. AB - Cool-season grasses are frequently infected by Neotyphodium endophytes and this association is often considered as a mutualistic symbiosis. We examined the incidence of Neotyphodium in populations of Bromus setifolius, Phleum alpinum and Poa spiciformis, native and wide-spread grasses from south Patagonia, Argentina. The incidence of 36 populations of Bromus setifolius was studied in association with climatic and soil variables. 31 populations of Ph. alpinum were sampled in five different plant communities. Seventeen populations of P. spiciformis were sampled in three different plant communities. The association between incidence and climatic variables in Ph. alpinum and between incidence and soil fertility in P. spiciformis was investigated. In B. setifolius endophyte incidence was positively correlated with annual average rainfall contrary to the results found in Ph. alpinum. All the populations of P. spiciformis were infected by endophytes and the incidence was associated with plant community. The Neotyphodium-grass interaction is variable in natural populations, supporting the increasing evidence that the Neotyphodium-host interaction depends, in many cases, on the environmental conditions. Field observations suggest that in detrimental low growth conditions the association is not favoured, leading to a decrease in the endophyte frequency of infection or even to the complete loss of the association. PMID- 17466028 TI - High-throughput culturing of fungi from plant litter by a dilution-to-extinction technique. AB - High-throughput bacterial cultivation has improved the recovery of slow-growing and previously uncultured bacteria. The most robust high-throughput methods are based on techniques of 'dilution to extinction' or 'extinction culturing'. The low-density partitioning of CFUs in tubes or microwells exploits the fact that the number of culturable species typically increases as inoculum density decreases. Bacterial high-throughput culturing methods were adapted to fungi to generate large numbers of fungal extinction cultures. The efficiency of extinction culturing was assessed by comparing it with particle filtration and automated plate-streaking. Equal volumes of particle suspension from five litter collections of the New Zealand forest tree Elaeocarpus dentatus were compared. Dilute particle suspensions of litter were pipetted into 48-well tissue culture plates containing 1 mL of agar medium per well. Particle volumes from the same samples were applied to continuous agar surfaces in Omnitray plates by automated streaking, and fungal diversity and richness were measured. The spectrum of isolates was assessed by microscopy and sequencing of the ITS or 28S region of the rRNA gene. Estimates of species diversity between the two methods were comparable, but extinction culturing increased species richness. Compared with plating methods using continuous surfaces, extinction culturing distributes fungal propagules over partitioned surfaces. Intercolony interactions are reduced, permitting longer incubation times, and colony initiation and recovery improved. Effort to evaluate and recover colonies from fungal isolation plates was substantially reduced. PMID- 17466029 TI - Qualitative and quantitative changes of beech wood degraded by wood-rotting basidiomycetes monitored by Fourier transform infrared spectroscopic methods and multivariate data analysis. AB - Beech wood (Fagus sylvatica L.) veneers were cultivated with white and brown rot fungi for up to 10 weeks. Fungal wood modification was traced with Fourier transform near infrared (FT-NIR) and Fourier transform mid infrared (FT-MIR) methods. Partial least square regression (PLSR) models to predict the total lignin content before and after fungal decay in the range between 17.0% and 26.6% were developed for FT-MIR transmission spectra as well as for FT-NIR reflectance spectra. Weight loss of the decayed samples between 0% and 38.2% could be estimated from the wood surface using individual PLSR models for white rot and brown rot fungi, and from a model including samples subjected to both degradation types. PMID- 17466030 TI - Sequence analysis of the 181-kb accessory plasmid pSmeSM11b, isolated from a dominant Sinorhizobium meliloti strain identified during a long-term field release experiment. AB - The 181 251 bp accessory plasmid pSmeSM11b of Sinorhizobium meliloti strain SM11, belonging to a dominant indigenous S. meliloti subpopulation identified during a long-term field release experiment, was sequenced. This plasmid has 166 coding sequences (CDSs), 42% of which encode proteins with homology to proteins of known function. Plasmid pSmeSM11b is a member of the repABC replicon family and contains a large gene region coding for a conjugation system similar to that of other self-transmissible plasmids in Rhizobium and Agrobacterium. Another pSmeSM11b gene region, possibly involved in sugar metabolism and polysaccharide catabolism, resembled a region of S. meliloti 1021 megaplasmid pSymB and in the genome of Sinorhizobium medicae WSM419. Another module of plasmid pSmeSM11b encodes proteins similar to those of the nitrogen-fixing actinomycete Frankia CcI3, and which are likely to be involved in the synthesis of a secondary metabolite. Several ORFs of pSmeSM11b were predicted to play a role in nonribosomal peptide synthesis. Plasmid pSmeSM11b has many mobile genetic elements, which contribute to the mosaic composition of the plasmid. PMID- 17466031 TI - Ectomycorrhizal fungi: exploring the mycelial frontier. AB - Ectomycorrhizal (ECM) fungi form mutualistic symbioses with many tree species and are regarded as key organisms in nutrient and carbon cycles in forest ecosystems. Our appreciation of their roles in these processes is hampered by a lack of understanding of their soil-borne mycelial systems. These mycelia represent the vegetative thalli of ECM fungi that link carbon-yielding tree roots with soil nutrients, yet we remain largely ignorant of their distribution, dynamics and activities in forest soils. In this review we consider information derived from investigations of fruiting bodies, ECM root tips and laboratory-based microcosm studies, and conclude that these provide only limited insights into soil-borne ECM mycelial communities. Recent advances in understanding soil-borne mycelia of ECM fungi have arisen from the combined use of molecular technologies and novel field experimentation. These approaches have the potential to provide unprecedented insights into the functioning of ECM mycelia at the ecosystem level, particularly in the context of land-use changes and global climate change. PMID- 17466032 TI - Possible pitfall in diagnosis: mitotic arrest of gastric epithelium after docetaxel therapy for hormone-refractory prostatic cancer. PMID- 17466033 TI - Lesson of the month. Beware the eosinophils. PMID- 17466034 TI - Online tumor conference in the clinical management of gynecological cancer: experience from a pilot study in Germany. AB - The concept of the online tumor conference was established in 2004 as a pilot project. We developed specific web-based software to organize and conduct online tumor board meetings of gynecologists, surgeons, radiologists, oncologists, and pathologists from different hospitals and gynecological practitioners, discussing individual patient's cases, defining therapy options, and exchanging clinical experience. Following a didactic approach, patient data are presented to the participants, with a special focus toward patient's preference and late toxicity from prior therapy. Then different national (eg, Arbeitsgemeinschaft Gynaekologische Onkologie, Deutsche Gesellschag fur Gynaekologic und Geburtshilfe) and international guidelines (eg, American Society of Clinical Oncology, National Cancer Institute), current study results based on literature review and open clinical trials are discussed. An individual diagnosis and therapy recommendation for each patient is reached by consensus. All protocols, guidelines, and publication data are upgraded and dispersed via Internet for all participants. In the period from December 2004 to August 2006, 39 tumor board conferences were performed with a total of 667 participants. One hundred forty four patients' cases were presented, and 121 peer-reviewed second-opinions were sought. In an anonymous survey, 84% of the participants reported to be satisfied with the information content and 72% with the technical support. Overall 98% of the individual therapy recommendations were accepted and implemented. The tumor board conference presents an optimal possibility for extensive scientific discussions and exchange (92%) and improves advanced educational training (81%). In conclusion, the online tumor conference is feasible and represents a time saving possibility for gynecological oncologist to receive a treatment recommendation based on the best available clinical and scientific evidence. PMID- 17466035 TI - Silent killer of the night: a feasibility study of an outreach well-women clinic for cervical cancer screening in female sex workers in Hong Kong. AB - The purpose of this study was to see if an outreach model could be a feasible option for early detections of preinvasive cervical cancer in women with double identities of female sex workers (FSW) and illegal migrant workers in Hong Kong who most needed cervical screening but were often deprived of such a service. High turnover rates, acceptability, and compliance for follow-ups could potentially render such a clinical model unsuccessful. A total of 245 FSW were screened at the outreach clinic from January 2004 to December 2005, which was set up in a nongovernmental organization in a red light district. A questionnaire regarding their lifestyles and demographic details was used before a gynecological history, Papanicolaou (PAP) smear, and other health checkup were conducted. Chi-square test and multinomial logistic regression were used to analyze the results. Of 235 women tests, 9.8% of them had CIN I-III, and places of origin were found to be important risk factors for abnormal PAP smears. The nonlocal workers were significantly more likely to have abnormal PAP smears (chi(2)= 10.55, P= 0.04). Among the women, 88.1% of them who had the tests returned for follow-up with poorer compliance among those with an abnormal result. We conclude that an outreach well-women clinic seems to be an acceptable option for these women and an effective way for the early detection of cervical cancer. PMID- 17466036 TI - Loss of phosphatase and tensin homologue deleted on chromosome 10 and phosphorylation of mammalian target of rapamycin are associated with progesterone refractory endometrial hyperplasia. AB - The objective of our study was to evaluate the phosphatase and tensin homologue deleted on chromosome 10 (PTEN), p27, and mammalian target of rapamycin (mTOR) expressions in women with progesterone-responsive and refractory endometrial hyperplasia (EH) samples and to determine if these markers could be associated with response or used as potential targets for treatment. Thirty-eight matched pre- and posttreatment pairs of paraffin-embedded endometrial biopsies were obtained from patients with EH. Immunohistochemical analysis for PTEN, p27, and phospho-mTOR were performed on all samples. Median age at diagnosis was 49 years (20-79 years). Median treatment interval was 3 months (1-12 months). Sixteen patients (42.1%) had complete resolution of their hyperplasia (responders), and 22 (57.9%) had persistent hyperplasia (nonresponders) after treatment with progesterone. In the pretreatment samples, no markers were found to predict nonresponders. In posttreatment samples, loss of PTEN expression with phospho mTOR expression was observed in more nonresponders than responders (40.9% vs 6.3%; P= 0.03). Phospho-mTOR overexpression was found in 63.6% of nonresponders. We found that persistent hyperplasia refractory to progesterone therapy was associated both with the loss of PTEN and with the loss of phosphorylation of mTOR. In select cases of non-responsive progesterone refractory EH, a rational target for treatment may involve the mTOR pathway. PMID- 17466037 TI - Immunohistochemical evaluation of cell proliferation and apoptosis markers in ovaries and uterus of tamoxifen-treated rats. AB - The aim of the study was to evaluate the immunohistochemical expression of cell proliferation and apoptosis markers in the ovaries and uterus of tamoxifen treated rats. Twelve rats (150-200 g) were divided into two equal groups. The study group received daily intraperitoneal injections of tamoxifen dissolved in 5% dimethyl sulfoxide (n= 6). The control group received only the vehicle (n= 6). The rats were sacrificed at the 20th day of injection and were perfused. The ovaries and uterus of the rats were extracted. The sections were immunohistochemically stained with cell proliferation marker Ki-67 and the apoptosis markers PTEN and CD95. The expressions of the markers were quantified by a semiquantitative H-score method in myometrium, endometrial glands, ovarian surface epithelium, ovarian follicles, corpus luteum, and ovarian stroma separately. The mean H-scores of CD95 and PTEN obtained from myometrium, glandular endometrium, ovarian surface epithelium, ovarian follicles, corpus luteum, and ovarian stroma did not show significant difference between the study and the control groups. Proliferative index (Ki-67) of endometrial glands was significantly higher in the study group than in the control group (P < 0.05). In addition, proliferative index (Ki-67) of corpus luteum was significantly higher in the study group than in the control group (P < 0.05). Tamoxifen treatment has a potential to stimulate the cell proliferation of endometrial glands and corpus luteum in tamoxifen-treated rats. Apoptosis markers of PTEN and CD95 did not demonstrate significant difference after the tamoxifen treatment. PMID- 17466038 TI - Short-interfering RNA-mediated silencing of proliferating cell nuclear antigen inhibit proliferation and induce apoptosis in HeLa cells. AB - Proliferating cell nuclear antigen (PCNA) is an important protein for DNA polymerase delta in the nucleus, and shown to have a fundamental role in cellular proliferation. It is overexpressed to support cell growth in cervical carcinoma. To study its role in stress response, we design and use short hairpin RNA (shRNA) to inhibit PCNA expression in HeLa cells and validate its effect on cell proliferation. In this study, three PCNA-shRNA expression vectors are constructed and introduced into HeLa cells, and the cell cycle is analyzed by flow cytometry. Apoptotic cell is detected by single cell gel electrophoresis assay (comet assay), and caspase cleavage is studied also. Expression of PCNA is assessed by real-time reverse transcription-polymerase chain reaction and Western blot analysis. Upon transient transfection with plasmid encoding shRNA, it is found that expression of PCNA decreased in shRNA-transfected cells, downregulation of PCNA inhibit cell growth and induce apoptosis in HeLa cells. PCNA downregulation also increase cell population in the G0-G1 phase. In conclusion, our findings demonstrate that shRNA can inhibit the DNA replication and induce apoptosis in HeLa cells effectively and, therefore, could be used as a new potential anticancer tool for therapy of human cervical carcinoma. PMID- 17466039 TI - Does fecal diversion offer any chance for spontaneous closure of the radiation induced rectovaginal fistula? AB - Analysis of the clinical course of patients with postirradiation rectovaginal fistula after fecal diversion. The studied group included 17 women with postirradiation rectovaginal fistula who underwent fecal diversion as a sole mode of treatment, between January 1987 and December 2002, in our department. All patients were subjected to radiotherapy due to cancer of the uterine cervix, administered 5-107 months before the fistula appearance (mean, 22.9 months). In 3 of 17 patients (18%), spontaneous closure of fistula was observed after 5, 6, and 9 months, respectively, from fecal diversion. Closure was confirmed by endoscopy. Length of follow-up after fecal diversion ranged from 0.5 to 122 months. The actuarial probability of spontaneous closure of postradiotherapy rectovaginal fistula was 0.24 at 9 months of follow-up and then remained stable thereafter. In conclusion, colostomy alone gives hardly a chance for closure of the postradiotherapy rectovaginal fistula. Additional surgical measures are necessary. PMID- 17466040 TI - p16(CDKN2) gene polymorphism: association with histologic subtypes of epithelial ovarian cancer in China. AB - p16 is an important tumor suppressor gene, which is inactivated in many kinds of tumors. The common variants of p16 may be associated with the risk of certain tumors development. We analyzed the frequency of two adjacent polymorphisms in p16 exon 3 (540C-->G and 580C-->T) and their haplotype in blood samples from epithelial ovarian cancer (EOC) patients and healthy controls using polymerase chain reaction-restriction fragment length polymorphism. The results showed that the genotype frequency of p16 580C-->T polymorphism was significantly different among histologic subtypes of EOC (P= 0.02). T allele carriers significantly reduced the risk of serous EOC; the adjusted odds ratio was 0.40 (95% CI = 0.19 0.84). There are neither association between p16 540C-->G polymorphism and EOC development, progression, nor association between the haplotypes of two single nucleotide polymorphisms and the tumor development. Our results suggested that the p16 580C-->T polymorphism might affect the individual susceptibility to specific subtypes of EOC. Different types of ovarian cancer might adopt distinct carcinogenetic pathways. However, this result may be further validated in a larger sample of patients. PMID- 17466041 TI - A case of ovarian endometrioid adenocarcinoma with a yolk sac tumor component. AB - Endometrioid adenocarcinoma of the ovary coexists very rarely with yolk sac tumor (YST). This unusual mixed tumor is thought to be a rare variant of endometrioid ovarian carcinoma because of its aggressive behavior, lack of response to chemotherapy, and unfavorable prognosis. We report a case of ovarian endometrioid adenocarcinoma with a YST component in a postmenopausal woman. The patient was treated by surgery and a combination of bleomycin, etoposide, and cisplatin and taxol and carboplatin. She has been clinically free of tumor for 20 months. Immunohistochemically, the YST component reacted for alpha-fetoprotein. YST areas were negative for both CA125 and sex-hormone receptors. Cytokeratin7 and epithelial membrane antigen were negative in YST, but positive in endometrioid adenocarcinoma. The occurrence of this unusual case suggests that even somatic carcinomas may acquire an extraembryonal germ cell differentiation. PMID- 17466042 TI - Follow-up strategies in gynecological oncology: searching appropriateness. AB - Nowadays, the cost for oncology diseases is growing rapidly, in particular as a consequence of the introduction of new drugs and new diagnostic procedures, and becoming a considerable percentage of the global healthcare expense. On the other hand, a substantial amount of that cost is considered to be imputable to the follow-up procedures. The aim of our paper is to introduce the debate about follow-up policies adopted in gynecological oncology throughout a literature review just based on cost-effectiveness and cost-efficacy in order to explore if the data are consistent with evidences available in this field. Furthermore, it is discussed if common practice fits the needs of patients, gynecological oncologists, and health service. Despite the fact that in gynecological oncology we must consider different clinical situations concerning each specific neoplasm with their peculiar natural history, some general considerations could be drawn in order to set up future initiatives properly. PMID- 17466043 TI - The effect of epithelial and stromal tumor components on FIGO stages III and IV ovarian carcinosarcomas treated with primary surgery and chemotherapy. AB - The aim of this study is to assess the effect of epithelial and stromal tumor components on survival outcomes in FIGO stage III or IV ovarian carcinosarcomas (OCS) treated with primary surgery and adjuvant chemotherapy at the Northern Gynaecological Oncology Centre (NGOC), Gateshead. Women were identified from the histopathology/NGOC databases. Age, FIGO stage, details of histology, treatment, and overall survival were recorded. Of 34 cases (1994-2006, all FIGO stages), 17 were treated with primary surgery followed by adjuvant chemotherapy for FIGO stage III or IV. The median age was 66 years (52-85 years). Cytoreduction was optimal (n= 9) or complete (n= 1) in 10/17 (59%) cases. Epithelial predominant (EP) or stromal predominant (SP) tumor (defined as >50% of either component in the primary tumor) was noted in 12 and 5 cases, respectively. Epithelial types included serous (n= 9), endometrioid (n= 5), and mixed types (n= 3). Twelve women have died of disease. The median overall survival was 11.0 months (3-74 months). On univariate analysis, survival was not affected by optimal/suboptimal debulking, platinum/doxorubicin-containing chemotherapy, or homologous/heterologous stromal components. Stromal components (>25%) adversely affected survival (P= 0.02), and there was a trend to worse survival with serous compared with nonserous epithelial components (P= 0.07). Cox regression (multivariate analysis) showed that SP tumors (P= 0.04), suboptimal debulking (P= 0.01), age (P= 0.01), and tumors with serous epithelial component (P= 0.05) were adverse independent prognostic factors. Type of chemotherapy and homologous/heterologous components (P= 0.24) did not affect overall survival. In conclusion, our study suggests that SP-OCS have a worse survival outcome than EP tumors. Tumors with serous epithelial components adversely affected the survival compared with nonserous components. Larger studies are required to confirm these effects and to identify the optimum chemotherapy regimen for OCS. PMID- 17466044 TI - Results of chemotherapy for pulmonary metastases of carcinoma of the cervix in patients after primary surgical and radiotherapeutic management. AB - The purpose of this study was to evaluate the results of chemotherapy of pulmonary metastases from invasive carcinoma of the cervix, which were detected after a disease-free period after initial treatment with surgery or radiotherapy. Fifty patients with radiologically proven pulmonary metastases were treated with chemotherapy. All patients received a platinum-5-fluorouracil (PF) program: cisplatin 75 mg/m(2) and 5-fluorouracil 800 mg/m(2) every 4 weeks. The overall 1- and 3-year survival after PF chemotherapy was 62% and 17.6%, respectively. The progression-free survival at 1 and 3 years was 36.7% and 14.3%, respectively. There were 6 (12%) complete responses and 17 (34%) partial responses. Hematologic tolerance was acceptable. Third degree and fourth degree leukopenia was diagnosed in four (8%) and six (12%) patients, respectively. Three individuals had third degree thrombocytopenia. In the multivariate analysis, the following prognostic factors were associated with poor survival: time to recurrence after primary treatment (P= 0.002), number of lung metastases (P= 0.016), and progression during chemotherapy (P= 0.001). We conclude that PF regimen is a safe and reasonably effective chemotherapy in the management of patients with pulmonary metastases after primary treatment for invasive carcinoma of the cervix who do not qualify for surgical metastasectomy. PMID- 17466045 TI - Carboplatin chemotherapy in a pregnant patient with undifferentiated ovarian carcinoma: case report and review of the literature. AB - There are already 12 reports of women treated by chemotherapy for epithelial ovarian cancer during pregnancy. However, most cases received chemotherapy of single cisplatin or cisplatin-based regime, and only four cases received carboplatin-containing chemotherapy. We report the case of a woman treated with single-agent carboplatin during pregnancy. The patient underwent bilateral salpingo-oophorectomy at 18 weeks of gestation and was diagnosed as having stage IC undifferentiated ovarian carcinoma. She was treated with four courses of carboplatin (area under the curve = 6.0) chemotherapy during pregnancy without severe toxicity. At 33 weeks of gestation, cesarean section was performed, followed by total hysterectomy, omentectomy, and pelvic and para-aortic lymphadenectomy. No residual disease was histologically shown. The patient underwent additional chemotherapy with carboplatin and paclitaxel. After one year of follow-up, the baby shows normal growth and the patient has no evidence of disease. Postponing the termination of pregnancy by single-agent carboplatin chemotherapy during pregnancy might be considered as an option for therapy in selected women with ovarian malignancies. PMID- 17466046 TI - Vesicouterine ligament contains abundant autonomic nerve ganglion cells: the distribution in histology concerning nerve-sparing radical hysterectomy. AB - The aim of this study is to describe the histologic architecture of the tissues corresponding to the surgically developed connective tissue bundle commonly referred to as the posterior leaf of the vesico-uterine ligament (VUL), and to examine distribution of ganglion cells. Serial macroscopic slices, each 15-20 mm in thickness, were made from eight specimens (obtained from six female elderly cadavers). In these macroslices, the location of the deep uterine vein was used to identify the deep leaf of the VUL. The specimens were trimmed and semi-serial histologic sections in thickness were prepared at 1 mm intervals. Vesical veins and the associated nerve elements were enclosed by fascia and formed a common pedicle. The base of the pedicle contained the deep uterine vein trunk. The fascia encircling the pedicle varied in thickness and connective intensity between specimens. This vesical neurovascular bundle contained abundant ganglion cells. On average, 48.0% of the ganglion cells along the vesical tributaries of the deep uterine vein were located on the medial or vaginal side of the veins, 19.2% were located between veins, 13.0% on the lateral side of the veins, and 19.8% on the dorsal side. The interindividual variability was greatest on the dorsal side of vesical veins and ranged 11-202 cells. We conclude that in order to achieve maximal preservation of the ganglion cells during the surgical dissection of the posterior leaf of the VUL, care must be taken when the medial or vesical aspect of the ligament is separated. The standard nerve-sparing radical hysterectomy should be modified to reflect differences in the distribution of ganglion cells and in connective intensity between ganglions and veins. PMID- 17466047 TI - A review of the close surveillance policy for stage I female germ cell tumors of the ovary and other sites. AB - Ovarian germ cell tumors are rare but very curable at all stages of disease. There is good evidence that surveillance for stage I dysgerminomas is a safe option although many centers worldwide still advocate adjuvant chemotherapy for stage IA nondysgerminomatous tumors, despite the significant risk of developing long-term treatment side effects. Here, we review the safety of our ongoing surveillance program of all stage IA female germ cell tumors. Thirty-seven patients (median age 26, range 14-48 years) with stage I disease were referred to Mount Vernon and Charing Cross Hospitals between 1981 and 2003. Patients underwent surgery and staging followed by intense surveillance, which included regular tumor markers and imaging. The median period of follow-up was 6 years. Relapse rates for stage IA nondysgerminomatous tumors and dysgerminomas were 8 of 22 (36%) and 2 of 9 (22%), respectively, plus one patient with mature teratoma and glial implants also relapsed; 10 of these 11 patients (91%) were successfully cured with platinum-based chemotherapy. Only one patient died from chemoresistant disease. All relapses occurred within 13 months of initial surgery. The overall disease-specific survival of malignant ovarian germ cell tumors was 94%. Over 50% of patients who underwent fertility-sparing surgery went on to have successful pregnancies. We have confirmed again that surveillance of all stage IA ovarian germ cell tumors is very safe and that the outcome is comparable with testicular tumors. We question the need for potentially toxic adjuvant chemotherapy in nondysgerminoma patients who have greater than 90% chance of being salvaged with chemotherapy if they relapse later. PMID- 17466048 TI - Distribution of human papillomavirus genotypes in the patients with cervical carcinoma and its precursors in Zhejiang Province, China. AB - Various evidences reveal that the human papillomavirus (HPV) is the single most important etiologic agent in cervical carcinoma (CC). To investigate the distribution of HPV genotypes in the patients with CC and its precursors in Zhejiang Province, China, a total of 631 eligible samples from patients in Zhejiang Province with CC (N = 181), cervical intraepithelial neoplasia (CIN) II III (N = 345), and CIN I (N = 105) were detected. Age-matched samples of 217 women without cervical neoplasia were detected as control. An improved polymerase chain reaction (PCR)-restriction fragment length polymorphism assay validated by Hybrid Capture II and PCR sequencing was designed for HPV genotype. The prevalence of HPV was 95.0% in CC, 88.4% in CIN II-III, and 73.3% in CIN I, while only 41.9% in control. High-risk/low-risk HPV ratio showed a significant trend of increase with increased grade of CIN and transformation to carcinoma. A total of 24 HPV genotypes were detected in CC and its precursors. Of those, HPV 16 (65.2%), 18 (9.4%), and 58 (9.4%) were the major HPV genotypes in CC, while HPV 16 (37.6%), 58 (19.1%), 33 (10.6%), and 18 (5.4%) in CIN. In conclusion, the distribution of predominant HPV genotypes in the patients with CC and its precursors in Zhejiang Province is HPV 16, 58, and 18, probably as well as 33, which may be high-risk factors for CC. PMID- 17466049 TI - Chemoradiation for invasive cervical cancer in elderly patients: outcomes and morbidity. AB - Age may impact survival and treatment in cervical cancer patients. We sought to determine if treatment and survival were different in elderly patients and whether chemoradiation increased morbidity. We performed a retrospective chart review to identify patients treated with definitive radiation therapy at the University of Iowa Hospitals and Clinics between 1997 and 2001. Three hundred sixty-four patients had a new diagnosis of invasive cervical cancer, of which 150 patients were treated with radiation. We excluded patients treated postoperatively or with palliative intent, leaving 96 patients treated with definitive radiation therapy. Patients were divided into two age categories: elderly (>/=65) and nonelderly (<65). We compared these groups with respect to treatment received, morbidity, and survival. Sixty-nine (72%) women were less than 65 years old, and 27 (28%) women were greater than or equal to 65 years old. Chemoradiation was associated with decreased mortality (P < 0.01). The decrease in mortality did not differ between the two age cohorts (all causes: P = 0.66; cancer specific: P = 0.65), nor was there a difference in the complication rate due to chemoradiation (P = 0.70). Although elderly patients were more likely to be diagnosed with nonsquamous histologies (P < 0.01), their odds of receiving chemoradiation were 0.35 (95% CI: 0.13-0.90) times the odds for nonelderly. Elderly cervical cancer patients more often have nonsquamous histology and are likely to receive only radiation therapy compared to younger patients. Treatment with chemoradiation was associated with similar survival increases in both age cohorts. Complication rates between the two were similar. Chemoradiation should be considered in elderly patients with invasive cervical cancer. PMID- 17466050 TI - Expression of receptor-binding cancer antigen expressed on SiSo cells and estrogen receptor subtypes in the normal, hyperplastic, and carcinomatous endometrium. AB - The objectives were to study the expression of receptor-binding cancer antigen expressed on SiSo cells (RCAS1) and estrogen receptor (ER) subtypes in the normal, hyperplastic, and carcinomatous endometrium and to explore their possible role in carcinogenesis and progression of endometrial carcinoma. Immunohistochemistry and semiquantitative reverse transcriptase-polymerase chain reaction (RT-PCR) were applied to detect protein and messenger RNA expression of RCAS1, ER-alpha, and ER-beta in normal, hyperplastic, and carcinomatous endometrium. Western blotting was also used to detect the RCAS1 protein expression. Immunohistochemistry showed that the high expressions of RCAS1 protein were 0% (0/20), 9.1% (2/22), 40% (8/20), and 68.0% (34/50) in normal, simple, and complex hyperplasia, atypical hyperplasia, and endometrial carcinoma, respectively. There was a significant difference between each group (P < 0.05). The high-level expression of RCAS1 was detected more frequently in endometrial cancer with deep myometrial invasion, vascular invasion, and positive ER-alpha (P < 0.05). Two staining patterns of RCAS1 were observed. All normal, simple, and complex hyperplastic endometrium showed P pattern, while all malignant endometrium were of the D pattern. In atypical endometrium, 25% (5/20) cases showed D pattern. The Western blotting and RT-PCR results correlated with the immunohistochemistry results. The expression and distribution of RCAS1 may be involved in the malignant transformation of endometrium, and RCAS1 coexpression with ER-alpha may be associated with development and metastasis of endometrial carcinoma. PMID- 17466051 TI - Correlation between patient weight and defects in DNA mismatch repair: is this the link between an increased risk of previous cancer in thinner women with endometrial cancer? AB - The objective is to determine the relationship between obesity and defects in DNA mismatch repair (MMR) in women with endometrial cancer and to establish whether our previous finding of a higher rate of previous malignancy in thinner women with endometrial cancer is related to these factors. Specimens from 109 patients with primary uterine cancer were used to create a tissue microarray, which was stained with antibodies against MMR genes MLH1, MSH2, MSH6, and PMS2. Genotyping of normal and tumor tissues for microsatellite instability (MSI) was performed. Patients were stratified by body mass index (BMI) and correlated with a history of previous malignancy and defects in MMR. The average BMI of the overall population was 33 kg/m(2). Defective MMR was seen in 22% of tumors. The mean BMI in patients with tumors with MSI was 30.5, compared with 33.8 in microsatellite stable (MSS) tumors (P= 0.06); MSS tumors were more commonly seen in patients with a BMI more than 40 (25% vs 5% in patients with tumors with MSI, P= 0.07). Prior to their diagnosis of endometrial cancer, 16/109 (15%) patients reported having a prior malignancy, 11 (69%) had breast cancer, and 1 had colorectal cancer. Patients with tumors with MSI had previous cancer in 17% of cases, compared with 14% of patients with MSS tumors (P= 0.75). Our previous finding of an increased rate of prior malignancy in thinner patients with endometrial cancer does not appear to be due to alterations in MMR, and hereditary nonpolyposis colorectal cancer-associated cancers are rarely the prior malignancy. PMID- 17466052 TI - Omental metastases in clinical stage I endometrioid adenocarcinoma. AB - The clinical benefit of an omentectomy in endometrioid adenocarcinoma is unclear. The objective of this study was to clarify the significance of an omentectomy performed for clinical stage I endometrioid adenocarcinoma. A prospective study was performed on 134 patients with clinical stage I endometrioid adenocarcinoma who underwent omentectomy in addition to a staging laparotomy between 1998 and 2004: simple total hysterectomy, bilateral salpingo-oophorectomy, pelvic and para aortic lymph node dissection, and peritoneal cytology. The frequency and prognosis of omental metastases and their relationships with extrauterine spread to other sites were investigated. Omental metastasis was noted in four patients (3.0%). As for extrauterine spread, the positivity rate of lymph node metastases was 13/128 (10.2%), peritoneal cytology was 13/133 (9.8%), and adnexal metastases was 10/134 (7.5%). Omental metastases correlated with peritoneal cytology and adnexal metastases (P < 0.05 for both); however, two of the omental metastases positive patients were peritoneal cytology negative. All omental metastases positive patients died shortly after surgery, showing that their prognosis was poor. The omental metastases rate for clinical stage I endometrioid adenocarcinoma was lower than the positive rates for extrauterine spread to other sites; thus, the routine application of omentectomy as a part of a staging laparotomy may not be efficacious. However, omental metastases are a significant poor prognostic factor, and intraoperative examination of the omentum by close inspection and palpation as well as pathologic examination, if possible, may be indicated. PMID- 17466053 TI - Concurrent daily cisplatin and extended-field radiation therapy for carcinoma of the cervix. AB - The aim of this study was to assess acute toxicities of concurrent low-dose daily cisplatin and extended-field radiation therapy (EFRT) for carcinoma of the uterine cervix. Fifteen women with cervical cancer who were treated with concurrent daily low-dose cisplatin and EFRT were analyzed. Daily cisplatin dose was fixed to 8 mg/m(2), which was determined in the preceding phase I study using pelvic radiotherapy. Twelve patients underwent either combined external beam radiation therapy and intracavitary brachytherapy or external beam radiation therapy alone. Three other patients were treated with adjuvant chemoradiotherapy after surgery. A total dose of EFRT ranged from 40 to 45 Gy, with an additional boost to the gross tumor volume up to 50.4-55 Gy. A median total dose of cisplatin during entire radiation therapy course was 224 mg/m(2) (range, 200-240 mg/m(2)). In 14 of 15 patients (93%), daily cisplatin could be delivered continuously as planned without any modification. Administration of cisplatin had to be interrupted in only one patient for only 3 days. Fourteen patients developed grade 2 or worse leukopenia including five after treatment, grade 2 in four, grade 3 in eight, and grade 4 in two. Grade 3 thrombocytopenia was observed in three patients. Grade 2 or worse anemia was observed in 12. Three patients had grade 3 nonhematologic toxicities, diarrhea in two, and nausea/vomiting in one. Although moderate to severe hematologic toxicities are common, this study suggests that concurrent low-dose daily cisplatin and EFRT are feasible. A cumulative cisplatin dose of greater than 200 mg/m(2) during radiation therapy could be achieved by using daily cisplatin dose of 8 mg/m(2). PMID- 17466054 TI - Human papillomavirus type distribution in women from Asia: a meta-analysis. AB - The aim of this study was to determine human papillomavirus (HPV) type distribution in women with and without cervical neoplasia from Asia and to estimate the potential future impact of an HPV 16/18 prophylactic vaccine in this region. A meta-analysis was conducted including 79 studies using polymerase chain reaction to detect HPV types. A total of 5954, 1653, 958, and 16,803 women with invasive cervical cancer (ICC), high-grade squamous intraepithelial lesions (HSIL), low-grade squamous intraepithelial lesions (LSIL), and normal cytology or histology were included, respectively. Type-specific prevalence of HPV types 6, 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66, 68, 70, 73, and 82 were estimated and stratified by cervical lesion grade. Overall HPV prevalence was 85.9%, 81.0%, 72.9%, and 14.4%, respectively, in women with ICC, HSIL, LSIL, and normal cytology/histology. In ICC, HPV 16 was the predominant type (52.4%), followed by HPV 18, 58, 33, 52, 45, 31, and 35. The estimated HPV 16/18-positive fraction was 66.9%, 40.4%, 26.7%, and 3.3% in women with ICC, HSIL, LSIL, and normal cytology or histology, respectively. In ICC, the estimated HPV 16/18 positive fraction was about 70% in all Asian geographic regions, with the exception of Japan (51.3%). HPV 16/18 vaccines are estimated to provide about 67% protection against ICC in Asia. HPV 58 and 52 were among the five most common types in ICC in eastern and southeastern Asia but not in south central Asia. After HPV 16 and 18, the next most six common HPV types were 58, 33, 52, 45, 31, and 35 that accounted for additional 20% of cervical cancer cases in Asia. For optimal population coverage, these HPV carcinogenic types should be considered for second-generation HPV prophylactic vaccines. PMID- 17466055 TI - A p38 MAP kinase regulates the expression of the Aedes aegypti defensin gene in mosquito cells. AB - An Aedes aegypti p38 (Aap38) mitogen-activated protein kinase was isolated and characterized in this study. The 1761 bp long full-length Aap38 cDNA encodes an open reading frame of 358 amino acids, exhibiting characteristics of Thr/Tyr dual kinase specificities. We showed that bacteria activate both the kinase activity of Aap38 and the expression of the Aedes aegypti defensin A (AaDefA) gene, which is inhibited by a p38 kinase inhibitor SB203580 and dsRNA interference of Aap38. A similar result was obtained by a reporter construct containing the AaDefA regulatory region linked to Ds-Red. The lipopolysaccharide-activated reporter gene was inhibited by SB203580. In addition, Aap38 translocated to the nucleus after lipopolysaccharide induction. Our findings suggest that the p38 protein kinase pathway is involved in the antibacterial peptide synthesis in mosquitoes. PMID- 17466058 TI - Penile rehabilitation after radical prostatectomy: where do we stand and where are we going? AB - INTRODUCTION: Postprostatectomy erectile dysfunction (ED) remains a serious quality-of-life issue. Recent advances in the understanding of the mechanism of postprostatectomy ED have stimulated great attention toward penile rehabilitation. AIM: This review presents and analyzes a contemporary series of the recent medical literature pertaining to penile rehabilitation therapy after radical prostatectomy (RP). MAIN OUTCOME MEASURES: The laboratory and clinical studies related to penile rehabilitation are analyzed. The validity of the methodology and the conclusion of the findings from each study are determined. METHODS: The published and presented reports dealing with penile rehabilitation following RP in human and cavernous nerve injury in animal models are reviewed. RESULTS: Exciting scientific discoveries have improved our understanding of postprostatectomy ED at the molecular level. The rationale for postprostatectomy penile rehabilitation appears to be logical according to animal studies. However, clinical studies have not consistently replicated the beneficial effects found in the laboratory studies. Currently available clinical studies are flawed due to short-term follow-up, small number of patients in the studies, studies with retrospective nature, or prospective studies without control. Rehabilitation programs are also facing a challenge with the compliance, which is critical for success for any rehabilitation program. At the present time, we do not have concrete evidence to recommend what, when, how long, and how often a particular penile rehabilitative therapy can be used effectively. CONCLUSIONS: Large prospective, multicentered, placebo-controlled trials with adequate follow-up are necessary to determine the cost-effective and therapeutic benefits of particular penile rehabilitative therapy or therapies in patients following the treatment of clinically localized prostate cancer. Until such evidence is available, it is difficult to recommend any particular penile rehabilitation program as a standard of practice. PMID- 17466059 TI - Confirmatory factor analysis of the German version of the international index of erectile function (IIEF): a comparison of four models. AB - INTRODUCTION: The International Index of Erectile Function (IIEF) is the most widely used instrument to assess erectile function in research and clinical practice. However, there are heterogeneous results concerning the factor structure of this questionnaire. The original model assumes five factors (erectile function, intercourse satisfaction, orgasmic function, sexual desire, and overall satisfaction). Others suggested four factors (composite domain of erectile dysfunction and intercourse satisfaction, orgasmic function, sexual desire, and overall satisfaction) or only two factors (sexual function and sexual desire). Because of the high intercorrelation between the different domains, a one-factor model is also plausible. AIMS: This study assesses and compares these four models of the German version of the IIEF. METHODS: It was examined which of the models fit best our data from 261 German men in cardiovascular rehabilitation participating in the SPARK study (Sexuality of Patients in the Rehabilitation of Cardiovascular Diseases). Contrary to the former exploratory studies, we used confirmatory factor analysis. MAIN OUTCOME MEASURES: Local and global goodness-of fit measures were calculated. RESULTS: The results show that two items (ability to maintain erection and intercourse frequency) could not be represented sufficiently through any of the four models. Based on the global goodness-of-fit indexes, our data proved to be fairly congruent with the original five-factor model and were acceptably represented by a four-factor model as well. CONCLUSIONS: The original five-factor structure could be confirmed. Due to high intercorrelations, the different domains cannot optimally be discriminated and should be interpreted with caution. Further research is needed to clarify the association between the domains of male sexual function. PMID- 17466060 TI - Risk factors for acute respiratory tract infections in general practitioner patients in The Netherlands: a case-control study. AB - BACKGROUND: Acute respiratory tract infections (ARTI) are an important public health problem. Improved identification of risk factors might enable targeted intervention. Therefore we carried out a case-control study with the aim of identifying environmental risk factors for ARTI consultations in the Dutch general population. METHODS: A subset of patients visiting their GP in the period of 2000-2003 with an ARTI (cases) and age-matched controls (visiting for other complaints) were included in a case-control study. They were asked to complete a questionnaire about potential risk factors. Conditional logistic regression was used to calculate odds ratio's (OR) and 95% confidence intervals (CI) to estimate the independent effect of potential risk factors. RESULTS: A total of 493 matched pairs of case and control subjects were enrolled. Exposure to persons with respiratory complaints, both inside and outside the household, was found to be an independent risk factor for visiting a GP with an ARTI (respectively ORadj = 1.9 and ORadj = 3.7). Participants exposed to dampness or mould at home (ORadj=0.5) were significantly less likely to visit their GP. In accordance with the general risk of consultations for ARTI, participants with a laboratory-confirmed ARTI who were exposed to persons with respiratory complaints outside the household were also significantly more likely to visit their GP (ORadj=2.5). CONCLUSION: This study confirmed that heterogeneity in the general population as well as in pathogens causing ARTI makes it complicated to detect associations between potential risk factors and respiratory infections. Whereas it may be difficult to intervene on the risk posed by exposure to persons with respiratory complaints, transmission of ARTI in the general population might be reduced by improved hygienic conditions. PMID- 17466061 TI - Molecular characterisation of the early response in pigs to experimental infection with Actinobacillus pleuropneumoniae using cDNA microarrays. AB - BACKGROUND: The bacterium Actinobacillus pleuropneumoniae is responsible for porcine pleuropneumonia, a widespread, highly contagious and often fatal respiratory disease of pigs. The general porcine innate immune response after A. pleuropneumoniae infection is still not clarified. The objective of this study was hence to characterise the transcriptional response, measured by using cDNA microarrays, in pigs 24 hours after experimental inoculation with A. pleuropneumoniae. METHODS: Microarray analyses were conducted to reveal genes being differentially expressed in inflamed versus non-inflamed lung tissue sampled from inoculated animals as well as in liver and tracheobronchial lymph node tissue sampled from three inoculated animals versus two non-inoculated animals. The lung samples were studied using a porcine cDNA microarray with 5375 unique PCR products while liver tissue and tracheobronchial lymph node tissue were hybridised to an expanded version of the porcine microarray with 26879 unique PCR products. RESULTS: A total of 357 genes differed significantly in expression between infected and non-infected lung tissue, 713 genes differed in expression in liver tissue from infected versus non-infected animals and 130 genes differed in expression in tracheobronchial lymph node tissue from infected versus non-infected animals. Among these genes, several have previously been described to be part of a general host response to infections encoding immune response related proteins. In inflamed lung tissue, genes encoding immune activating proteins and other pro-inflammatory mediators of the innate immune response were found to be up-regulated. Genes encoding different acute phase reactants were found to be differentially expressed in the liver. CONCLUSION: The obtained results are largely in accordance with previous studies of the mammalian immune response. Furthermore, a number of differentially expressed genes have not previously been associated with infection or are presently unidentified. Determination of their specific roles during infection may lead to a better understanding of innate immunity in pigs. Although additional work including more animals is clearly needed to elucidate host response to porcine pleuropneumonia, the results presented in this study demonstrate three subsets of genes consistently expressed at different levels depending upon infection status. PMID- 17466062 TI - Smoking among pregnant women in Cantabria (Spain): trend and determinants of smoking cessation. AB - BACKGROUND: Cantabria (Spain) has one of the highest prevalence of smoking among women of the European Union. The objectives are to assess the trend of smoking during pregnancy in a five-year period and the determinants of smoking cessation during pregnancy in Cantabria. METHODS: A 1/6 random sample of all women delivering at the reference hospital of the region for the period 1998-2002 was drawn, 1559 women. Information was obtained from personal interview, clinical chart, and prenatal care records. In the analysis relative risks and 95% confidence intervals were estimated. Multivariable analysis was carried out using stepwise logistic regression. RESULTS: Smoking prior to pregnancy decreased from 53.6% in 1998 to 39.4% in 2002. A decrease in smoking cessation among women smoking at the beginning of pregnancy was observed, from 37.3% in 1998 to 20.6% in 2002. The mean number of cigarettes/day (cig/d) before pregnancy remained constant, around 16 cig/d, whereas a slight trend to increase over time was seen, from 7.7 to 8.9 cig/d. In univariate analysis two variables favoured significantly smoking cessation, although they were not included in the stepwise logistic regression analysis, a higher education level and to be married. The logistic regression model included five significant predictors (also significant in univariate analysis): intensity of smoking, number of previous pregnancies, partner's smoking status, calendar year of study period (these four variables favoured smoking continuation), and adequate prenatal care (which increased smoking cessation). CONCLUSION: The frequency of smoking among pregnant women is very high in Cantabria. As smoking cessation rate has decreased over time, a change in prenatal care programme on smoking counseling is needed. Several determinants of smoking cessation, such as smoking before pregnancy and partner's smoking, should be also addressed by community programmes. PMID- 17466063 TI - Exploring the effect of changes to service provision on the use of unscheduled care in England: population surveys. AB - BACKGROUND: Unscheduled care is defined here as when someone seeks treatment or advice for a health problem without arranging to do so more than a day in advance. Recent health policy initiatives in England have focused on introducing new services such as NHS Direct and walk in centres into the unscheduled care system. This study used population surveys to explore the effect of these new services on the use of traditional providers of unscheduled care, and to improve understanding of help seeking behaviour within the system of unscheduled care. METHODS: Cross-sectional population postal surveys were undertaken annually over the five year period 1998 to 2002 in two geographical areas in England. Each year questionnaires were sent to 5000 members of the general population in each area. RESULTS: The response rate was 69% (33,602/48,883). Over the five year period 16% (5223/33,602) 95% CI (15.9 to 16.1) of respondents had an unscheduled episode in the previous four weeks and this remained stable over time (p = 0.170). There was an increased use of telephone help lines over the five years, reflecting the change in service provision (p = 0.008). However, there was no change in use of traditional services over this time period. Respondents were most likely to seek help from general practitioners (GPs), family and friends, and pharmacists, used by 9.0%, 7.2% and 6.3% respectively of the 5815 respondents in 2002. Most episodes involved contact with a single service only: 7.0% (2363/33,602) of the population had one contact and 2% (662/33,602) had three or more contacts per episode. GPs were the most frequent point of first contact with services. CONCLUSION: Introducing new services to the provision of unscheduled care did not affect the use of traditional services. A large majority of the population continued to turn to their GP for unscheduled health care. PMID- 17466064 TI - Inequity in the provision of and access to palliative care for cancer patients. Results from the Italian survey of the dying of cancer (ISDOC). AB - BACKGROUND: The palliative services and programs have been developed with different intensity and modalities in all countries. Several studies have reported that a geographic variation in the availability and provision of palliative care services between and within countries exists, and that a number of vulnerable groups are excluded from these services. This survey estimates the distribution of places of care for Italian cancer patients during the last three months of their lives, the proportion receiving palliative care support at home and in hospital, and the factors associated with the referral to palliative care services. METHODS: This is a mortality follow-back survey of 2,000 cancer deaths identified with a 2-stage probability sample, representative of the whole country. Information on patients' experience was gathered from the non professional caregiver through an interview, using an adapted version of the VOICES questionnaire. A section of the interview concerned the places of care and the palliative care services provided to patients. Multivariate logistic regression analyses were conducted to identify the determinants of palliative care service use. RESULTS: Valid interviews were obtained for 67% of the identified caregivers (n = 1,271). Most Italian cancer patients were cared for at home (91%) or in hospital (63%), but with substantial differences within the country. Only 14% of Italian cancer patients cared for at home against 20% of those admitted to hospital, received palliative care support. The principal determinants identified for receiving these service were: an extended interval between diagnosis and death (P = 0.01) and the caregiver's high educational level (P = 0.01) for patients at home; the low patient's age (P < 0.01) and the caregiver's high educational level (P = 0.01) for patients in hospital. CONCLUSION: In Italy palliative care services are not equally available across the country. Moreover, access to the palliative care services is strongly associated with socio demographic characteristics of the patients and their caregivers. Italian Policy-makers need to equalise palliative care provision and access across the country to meet the needs of all cancer patients. PMID- 17466065 TI - Control of interjoint coordination during the swing phase of normal gait at different speeds. AB - BACKGROUND: It has been suggested that the control of unconstrained movements is simplified via the imposition of a kinetic constraint that produces dynamic torques at each moving joint such that they are a linear function of a single motor command. The linear relationship between dynamic torques at each joint has been demonstrated for multijoint upper limb movements. The purpose of the current study was to test the applicability of such a control scheme to the unconstrained portion of the gait cycle--the swing phase. METHODS: Twenty-eight neurologically normal individuals walked along a track at three different speeds. Angular displacements and dynamic torques produced at each of the three lower limb joints (hip, knee and ankle) were calculated from segmental position data recorded during each trial. We employed principal component (PC) analysis to determine (1) the similarity of kinematic and kinetic time series at the ankle, knee and hip during the swing phase of gait, and (2) the effect of walking speed on the range of joint displacement and torque. RESULTS: The angular displacements of the three joints were accounted for by two PCs during the swing phase (Variance accounted for - PC1: 75.1 +/- 1.4%, PC2: 23.2 +/- 1.3%), whereas the dynamic joint torques were described by a single PC (Variance accounted for - PC1: 93.8 +/- 0.9%). Increases in walking speed were associated with increases in the range of motion and magnitude of torque at each joint although the ratio describing the relative magnitude of torque at each joint remained constant. CONCLUSION: Our results support the idea that the control of leg swing during gait is simplified in two ways: (1) the pattern of dynamic torque at each lower limb joint is produced by appropriately scaling a single motor command and (2) the magnitude of dynamic torque at all three joints can be specified with knowledge of the magnitude of torque at a single joint. Walking speed could therefore be altered by modifying a single value related to the magnitude of torque at one joint. PMID- 17466066 TI - Detection of mitochondrial insertions in the nucleus (NuMts) of Pleistocene and modern muskoxen. AB - BACKGROUND: Nuclear insertions of mitochondrial sequences (NuMts) have been identified in a wide variety of organisms. Trafficking of genetic material from the mitochondria to the nucleus has occurred frequently during mammalian evolution and can lead to the production of a large pool of sequences with varying degrees of homology to organellar mitochondrial DNA (mtDNA) sequences. This presents both opportunities and challenges for forensics, population genetics, evolutionary genetics, conservation biology and the study of DNA from ancient samples. Here we present a case in which difficulties in ascertaining the organellar mtDNA sequence from modern samples hindered their comparison to ancient DNA sequences. RESULTS: We obtained mitochondrial hypervariable region (HVR) sequences from six ancient samples of tundra muskox (Ovibos moschatus) that were reproducible but distinct from modern muskox sequences reported previously. Using the same PCR primers applied to the ancient specimens and the primers used to generate the modern muskox DNA sequences in a previous study, we failed to definitively identify the organellar sequence from the two modern muskox samples tested. Instead of anticipated sequence homogeneity, we obtained multiple unique sequences from both hair and blood of one modern specimen. Sequencing individual clones of a >1 kb PCR fragment from modern samples did not alleviate the problem as there was not a consistent match across the entire length of the sequences to Ovibos when compared to sequences in GenBank. CONCLUSION: In specific taxa, due to nuclear insertions some regions of the mitochondrial genome may not be useful for the characterization of modern or ancient DNA. PMID- 17466067 TI - Impact of parental history of substance use disorders on the clinical course of anxiety disorders. AB - BACKGROUND: Among the psychological difficulties seen in children of parents with substance use problems, the anxiety disorders are among the most chronic conditions. Although children of alcoholic parents often struggle with the effects of parental substance use problems long into adulthood, empirical investigations of the influence of parental substance use disorders on the course of anxiety disorders in adult offspring are rare. The purpose of this study was to examine prospectively the relationship between parental substance use disorders and the course of anxiety disorders in adulthood over the course of 12 years. METHODS: Data on 618 subjects were derived from the Harvard/Brown Anxiety Research Project (HARP), a longitudinal naturalistic investigation of the clinical course of multiple anxiety disorders. Kaplan-Meier survival estimates were used to calculate probabilities of time to anxiety disorder remission and relapse. Proportional hazards regressions were conducted to determine whether the likelihood of remission and relapse for specific anxiety disorders was lower for those who had a history of parental substance use disorders than for individuals without this parental history. RESULTS: Adults with a history of parental substance use disorders were significantly more likely to be divorced and to have a high school level of education. History of parental substance use disorder was a significant predictor of relapse of social phobia and panic disorders. CONCLUSION: These findings provide compelling evidence that adult children of parents with substance use disorders are more likely to have relapses of social phobia and panic disorders. Clinicians who treat adults with anxiety disorders should assess parental substance use disorders and dependence histories. Such information may facilitate treatment planning with regards to their patients' level of vulnerability to perceive scrutiny by others in social situations, and ability to maintain a long-term panic-free state. PMID- 17466068 TI - Individual differences in the use of the response scale determine valuations of hypothetical health states: an empirical study. AB - BACKGROUND: The effects of socio-demographic characteristics of the respondent, including age, on valuation scores of hypothetical health states remain inconclusive. Therefore, we analyzed data from a study designed to discriminate between the effects of respondents' age and time preference on valuations of health states to gain insight in the contribution of individual response patterns to the variance in valuation scores. METHODS: A total of 212 respondents from three age groups valued the same six hypothetical health states using three different methods: a Visual Analogue Scale (VAS) and two variants of the Time trade-off (TTO). Analyses included a generalizability study, principal components analysis, and cluster analysis. RESULTS: Valuation scores differed significantly, but not systematically, between valuation methods. A total of 36.8% of variance was explained by health states, 1.6% by the elicitation method, and 0.2% by age group. Individual differences in the use of the response scales (e.g. a tendency to give either high or low TTO scores, or a high or low scoring tendency on the VAS) were the main source of remaining variance. These response patterns were not related to age or other identifiable respondent characteristics. CONCLUSION: Individual response patterns in this study were more important determinants of TTO or VAS valuations of health states than age or other respondent characteristics measured. Further valuation research should focus on explaining individual response patterns as a possible key to understanding the determinants of health state valuations. PMID- 17466069 TI - The lin-35/Rb and RNAi pathways cooperate to regulate a key cell cycle transition in C. elegans. AB - BACKGROUND: The Retinoblastoma gene product (Rb) has been shown to regulate the transcription of key genes involved in cell growth and proliferation. Consistent with this, mutations in Rb are associated with numerous types of cancer making it a critical tumour suppressor gene. Its function is conferred through a large multiprotein complex that exhibits a dual function in both activation and repression of gene targets. In C. elegans, the Rb orthologue lin-35 functions redundantly with other transcriptional regulators to appropriately specify both vulval and pharyngeal cell fates. RESULTS: In C. elegans the intestinal cells must alter their cell cycle from the mitotic cell divisions typical of embryogenesis to karyokinesis and then endoreplication, which facilitates growth during larval development. While screening for genes that affect the ability of the intestinal cells to appropriately make this cell cycle transition during post embryonic development, we isolated mutants that either compromise this switch and remain mononucleate, or cause these cells to undergo multiple rounds of nuclear division. Among these mutants we identified a novel allele of lin-35/Rb, while we also found that the components of the synMuv B complex, which are involved in vulval specification, are also required to properly regulate the developmentally controlled cell cycle transition typical of these intestinal cells during larval development. More importantly, our work uncovered a role for certain members of the pathways involved in RNAi in mediating the efficient transition between these cell cycle programs, suggesting that lin-35/Rb cooperates with these RNAi components. Furthermore, our findings suggest that met-2, a methyltransferase as well as hpl-1 and hpl-2, two C. elegans homologues of the heterochromatin protein HP1 are also required for this transition. CONCLUSION: Our findings are consistent with lin-35/Rb, synMuv and RNAi components cooperating, probably through their additive effects on chromatin modification, to appropriately modulate the expression of genes that are required to switch from the karyokinesis cell cycle to endoreplication; a highly specified growth pathway in the intestinal epithelium. The lin-35/Rb repressor complex may be required to initiate this process, while components of the RNAi machinery positively reinforce this repression. PMID- 17466070 TI - Early efforts in modeling the incubation period of infectious diseases with an acute course of illness. AB - The incubation period of infectious diseases, the time from infection with a microorganism to onset of disease, is directly relevant to prevention and control. Since explicit models of the incubation period enhance our understanding of the spread of disease, previous classic studies were revisited, focusing on the modeling methods employed and paying particular attention to relatively unknown historical efforts. The earliest study on the incubation period of pandemic influenza was published in 1919, providing estimates of the incubation period of Spanish flu using the daily incidence on ships departing from several ports in Australia. Although the study explicitly dealt with an unknown time of exposure, the assumed periods of exposure, which had an equal probability of infection, were too long, and thus, likely resulted in slight underestimates of the incubation period. After the suggestion that the incubation period follows lognormal distribution, Japanese epidemiologists extended this assumption to estimates of the time of exposure during a point source outbreak. Although the reason why the incubation period of acute infectious diseases tends to reveal a right-skewed distribution has been explored several times, the validity of the lognormal assumption is yet to be fully clarified. At present, various different distributions are assumed, and the lack of validity in assuming lognormal distribution is particularly apparent in the case of slowly progressing diseases. The present paper indicates that (1) analysis using well-defined short periods of exposure with appropriate statistical methods is critical when the exact time of exposure is unknown, and (2) when assuming a specific distribution for the incubation period, comparisons using different distributions are needed in addition to estimations using different datasets, analyses of the determinants of incubation period, and an understanding of the underlying disease mechanisms. PMID- 17466071 TI - Effect of topical honey application along with intralesional injection of glucantime in the treatment of cutaneous leishmaniasis. AB - BACKGROUND: Leishmaniasis is an endemic disease in Iran. Although many treatments have been suggested for this disease, there hasn't been an effective and safe treatment yet. Regarding the healing effect of honey in the chronic ulcers and its reported therapeutic effect in cutaneous leishmaniasis, we performed a study to better evaluate the efficacy of honey in cutaneous leishmaniasis and its final scar. METHODS: In a prospective clinical trial, 100 patients with confirmed cutaneous leishmaniasis were selected and randomized into 2 groups. Group A were treated with topical honey twice daily along with intralesional injection of glucantime once weekly until complete healing of the ulcer or for maximum of 6 weeks. Group B were treated with intralesional injection of glucantime alone until complete healing of the ulcer or for a maximum of 6 weeks, too. The patients were followed for 4 months. The collected data were analyzed statistically using statistical tests including Chi-square, Mann Whitney and Kaplan-Mayer tests. RESULTS: In this study, 45 patients that had cutaneous leishmaniasis were treated with intralesional glucantime alone and 45 patients were treated with topical honey and glucantime. Ten patients left out the study. In the glucantime alone treated group, 32 patients (71.1%) had complete cure whereas in the group treated with both glucantime & topical honey, 23 patients (51.1%) achieved complete cure. This difference was significant statistically (p = 0.04). CONCLUSION: Further studies to better clarify the efficacy of honey in cutaneous leishmaniasis is needed. We suggest that in another study, the efficacy of honey with standardized level of antibacterial activity is evaluated against cutaneous leishmaniasis. PMID- 17466072 TI - Impairment of quality of life in parents of children and adolescents with pervasive developmental disorder. AB - BACKGROUND: Little is known about the Quality of Life (QOL) in parents of children with developmental diseases as compared to other severe neurological or psychiatric disorders. Aims of the present study were: to evaluate QOL in parents of children affected by Pervasive Development Disorder (PDDs), Cerebral Palsy (CP) or Mental Retardation (MR) as compared to a control group (CG); to evaluate QOL of parents of patients with different types of PDDs, namely Autistic Disorder (AD), High Function Autism/Asperger Syndromes (HFA/AS) and Pervasive Developmental Disorder Not Otherwise Specified (PPD-NOS); and to compare the level of impairment in QOL of mothers and fathers within PDDs, CP, MR groups and between AD, HFA/AS, PDD-NOS sub-groups. METHODS: The sample consisted of 212 parents (115 mothers and 97 fathers) of 135 children or adolescents affected by PDDs, MR or CP. An additional sample of 77 parents (42 mothers and 35 fathers) of 48 healthy children was also included and used as a control group. QOL was assessed by the WHOQOL-BREF questionnaire. RESULTS: Compared with parents of healthy children, parents in the PDDs group reported impairment in physical activity (p = 0.0001) and social relationships (p = 0.0001) and worse overall perception of their QOL (p = 0.0001) and health (p = 0.005). Scores in the physical (p = 0.0001), psychological (p = 0.0001) and social relationships domains (p = 0.0001) and in the physical (p = 0.0001) and social relationships (p = 0.0001) domains were lower compared to the MR group CP group respectively. Little differences were observed between MR, CP and control groups. The level of impairment of physical (p = 0.001) and psychological (p = 0.03) well-being were higher in mothers than in fathers in the PDDs and CP groups respectively; in the other groups, and across all the other domains of QQL impairment was similar. There were no statistically significant differences in the scores between the AD, HFA/AS and PDD-NOS sub-groups, but parents in the HFA/AS sub-group seemed to display a lower QOL compared to the AD sub-group. CONCLUSION: Parents of children with PDDs seem to display a higher burden, probably for a combination of environmental and genetic factors. Within this group of parents also those of HFA or AS people have higher stress. These finding must be taken into account in policy making to provide better and more specific supports and interventions for this group of diseases. PMID- 17466073 TI - Modelling evolution on design-by-contract predicts an origin of life through an abiotic double-stranded RNA world. AB - BACKGROUND: It is generally believed that life first evolved from single-stranded RNA (ssRNA) that both stored genetic information and catalyzed the reactions required for self-replication. PRESENTATION OF THE HYPOTHESIS: By modeling early genome evolution on the engineering paradigm design-by-contract, an alternative scenario is presented in which life started with the appearance of double stranded RNA (dsRNA) as an informational storage molecule while catalytic single stranded RNA was derived from this dsRNA template later in evolution. TESTING THE HYPOTHESIS: It was investigated whether this scenario could be implemented mechanistically by starting with abiotic processes. Double-stranded RNA could be formed abiotically by hybridization of oligoribonucleotides that are subsequently non-enzymatically ligated into a double-stranded chain. Thermal cycling driven by the diurnal temperature cycles could then replicate this dsRNA when strands of dsRNA separate and later rehybridize and ligate to reform dsRNA. A temperature dependent partial replication of specific regions of dsRNA could produce the first template-based generation of catalytic ssRNA, similar to the developmental gene transcription process. Replacement of these abiotic processes by enzymatic processes would guarantee functional continuity. Further transition from a dsRNA to a dsDNA world could be based on minor mutations in template and substrate recognition sites of an RNA polymerase and would leave all existing processes intact. IMPLICATIONS OF THE HYPOTHESIS: Modeling evolution on a design pattern, the 'dsRNA first' hypothesis can provide an alternative mechanistic evolutionary scenario for the origin of our genome that preserves functional continuity. REVIEWERS: This article was reviewed by Anthony Poole, Eugene Koonin and Eugene Shakhnovich. PMID- 17466074 TI - Genetic polymorphisms in dopamine-related genes and smoking cessation in women: a prospective cohort study. AB - BACKGROUND: Genes involved in dopaminergic neurotransmission have been suggested as candidates for involvement in smoking behavior. We hypothesized that alleles associated with reduced dopaminergic neurotransmission would be more common in continuing smokers than among women who quit smoking. METHODS: The study included 593 women aged 26-65 years who participated in a twelve month smoking cessation trial conducted in 1993-1994. Participants were contacted three years after the trial to obtain updated smoking history and biological specimens. Seven polymorphisms were assessed in genes involved in dopamine synthesis (tyrosine hydroxylase [TH]), receptor activation (dopamine receptors [DRD2, DRD3, DRD4]), reuptake (dopamine transporter [SLC6A3]), and metabolism (catechol-o methyltransferase [COMT]). Smoking cessation was assessed as "short-term" quitting (abstinence for the seven days before the conclusion of the trial) and "long-term" quitting (abstinence for the six months before a subsequent interview conducted several years later). RESULTS: We observed no association of any polymorphism with either short- or long-term quitting. Although some relative risk estimates were consistent with weak associations, either the direction of effect was opposite of that hypothesized, or results of the short- and long-term cessation endpoints differed. However, effect modification on smoking cessation was observed between DRD2 Taq1A and SLC6A3 VNTR polymorphisms, DRD3 Ser/Gly and d,1-fenfluramine, and DRD4 VNTR and d,1-fenfluramine. CONCLUSION: Although these results fail to support prior findings of independent associations of these polymorphisms with smoking status, our exploratory findings suggestive of gene gene and gene-treatment interactions warrants further investigation. PMID- 17466075 TI - Retroperitoneal Castleman's tumor and paraneoplastic pemphigus: report of a case and review of the literature. AB - BACKGROUND: Castleman's disease is a rare lymphoproliferative syndrome. Its etiology and pathogenesis are unclear. The disease can be occasionally associated with a paraneoplastic pemphigus (PNP), an autoimmune mucocutaneous disorder commonly seen in neoplasms of lymphocytic origin. CASE PRESENTATION: We present a case of a 63-year old male patient who was referred for surgical treatment of a lately diagnosed retroperitoneal pelvic mass. The patient had been already treated for two years due to progressive diffuse cutaneous lesions histologically consistent with lichen ruber verucosus and pemphigus vulgaris. Intraoperatively a highly vascularized solid mass occupying the small pelvis was resected after meticulous vascular ligation and hemostasis. After surgery and following immunosuppressive treatment a clear remission of the skin lesions was observed. CONCLUSION: Castleman's tumor should be always suspected when a retroperitoneal mass is combined with PNP. In a review of the literature we found 37 additional cases. Complete surgical resection of the tumor can be curative in most of the cases. PMID- 17466076 TI - Functional dissection of the ash2 and ash1 transcriptomes provides insights into the transcriptional basis of wing phenotypes and reveals conserved protein interactions. AB - BACKGROUND: The trithorax group (trxG) genes absent, small or homeotic discs 1 (ash1) and 2 (ash2) were isolated in a screen for mutants with abnormal imaginal discs. Mutations in either gene cause homeotic transformations but Hox genes are not their only targets. Although analysis of double mutants revealed that ash2 and ash1 mutations enhance each other's phenotypes, suggesting they are functionally related, it was shown that these proteins are subunits of distinct complexes. RESULTS: The analysis of wing imaginal disc transcriptomes from ash2 and ash1 mutants showed that they are highly similar. Functional annotation of regulated genes using Gene Ontology allowed identification of severely affected groups of genes that could be correlated to the wing phenotypes observed. Comparison of the differentially expressed genes with those from other genome wide analyses revealed similarities between ASH2 and Sin3A, suggesting a putative functional relationship. Coimmunoprecipitation studies and immunolocalization on polytene chromosomes demonstrated that ASH2 and Sin3A interact with HCF (host cell factor). The results of nucleosome western blots and clonal analysis indicated that ASH2 is necessary for trimethylation of the Lys4 on histone 3 (H3K4). CONCLUSION: The similarity between the transcriptomes of ash2 and ash1 mutants supports a model in which the two genes act together to maintain stable states of transcription. Like in humans, both ASH2 and Sin3A bind HCF. Finally, the reduction of H3K4 trimethylation in ash2 mutants is the first evidence in Drosophila regarding the molecular function of this trxG gene. PMID- 17466077 TI - The "Romsas in Motion" community intervention: program exposure and psychosocial mediated relationships to change in stages of change in physical activity. AB - BACKGROUND: Conducting process evaluations of health promoting interventions, and measuring the effectiveness of specific intervention components, may help in the understanding of program failure or success. The purposes of the present study were to examine adults' exposure to and involvement in specific components of a three year long pseudo-experimental community-based physical activity intervention, and to examine the relationship between such exposure and participation and changes in stages of change in physical activity and psychosocial mediators. METHODS: 1497 persons in the intervention group attended the baseline survey in 2000 (50.6%) and 1204 (80.4 of baseline attendees) provided data on the outcome variables of the present study. In 2003, 1089 were still living in the area, and were re-invited to follow-up assessments. Current analyses are based on the 603 persons (mean age 49 +/- 10 years) who provided baseline and follow-up data for the current purposes (56.6% follow-up rate). Process data, stages of change in physical activity, and potential psychosocial mediators of change in physical activity were assessed by questionnaires. The theory-based intervention was composed of communication, physical activity, environmental and participatory components. Data were analysed using frequency and descriptive statistics, Chi-square and t-tests, and regression analyses. RESULTS: Exposure and participation rates in the various intervention components varied greatly (1.5-92.7%). Participation in walking groups and aerobic exercise groups, as well as having seen the "Walk the stairs"-poster were significantly and positively related to change in stages of change in physical activity (beta = .12, p = .011; beta = .211, p < .001; beta = .105, p = .014, respectively). Additionally, having used the walk path was significantly and positively related to change in stages in women (beta = .209, p = .001) but not in men (beta = .011, p = .879), and in Western people (beta = .149, p = .003) but not in non Westerners (beta = -.293, p = .092). Observed significant relations were partly mediated by positive changes in psychosocial factors as social support from friends, perceived control, and physical activity identity. CONCLUSION: Findings revealed that particular intervention components, such as participation in physical activity groups, were more strongly related to forward transition in stages of change in physical activity than others. These findings together with results indicating that such transitions were mediated by specific psychosocial influences may improve theory and help to prioritize among specific intervention components in future programs. PMID- 17466078 TI - Growth arrest-specific protein 6 (GAS6) and the protein C pathway. PMID- 17466079 TI - Re-examining ethical obligations in the intensive care unit: HIV disclosure to surrogates. AB - Physicians treating newly incapacitated patients often must help navigate surrogate decision-makers through a difficult course of treatment decisions, while safeguarding the patient's autonomy. We offer guidance for intensive care physicians who must frequently address the difficult questions concerning disclosure of confidential information to surrogates. Three clinical vignettes will highlight the ethical challenges to physician disclosure of a critically ill patient's HIV status. Two key distinctions are offered that influence the propriety of disclosure: first, whether HIV infection represents a 'primary cause' for the patient's critical illness; and second, whether the surrogate may be harmed by failure to disclose HIV status. This balanced consideration of the direct duties of physicians to patients, and their indirect duties to surrogates and third-party contacts, may be used as a framework for considering other ethical obligations in the intensive care unit. We also provide a tabulation of individual US state laws relevant to disclosure of HIV status. PMID- 17466080 TI - Acute renal failure in patients with sepsis. PMID- 17466081 TI - Thrombelastography and sepsis. PMID- 17466082 TI - Hyperoxic acute lung injury and ventilator-induced/associated lung injury: new insights into intracellular signaling pathways. AB - In patients with acute respiratory distress syndrome (ARDS), supportive therapy with mechanical ventilation and oxygen is often life saving. Further acute lung injury however, is an unfortunate consequence of oxygen therapy as well as mechanical injury secondary to ventilator induced/associated lung injury (VI/ALI). In this issue of Critical Care, Li et al. expand on the intra-cellular signaling pathways regulating interactions between injury cascades resulting from hyperoxia and high tidal volume ventilation. The findings, suggest that interference or cooperation of different signals may have critical consequences as evidenced by indices of increased lung inflammation, microvascular permeability, and lung epithelial apoptotic cell death. PMID- 17466083 TI - An international initiative to identify genetic modifiers of cancer risk in BRCA1 and BRCA2 mutation carriers: the Consortium of Investigators of Modifiers of BRCA1 and BRCA2 (CIMBA). AB - BRCA1 and BRCA2 mutations exhibit variable penetrance that is likely to be accounted for, in part, by other genetic factors among carriers. However, studies aimed at identifying these factors have been limited in size and statistical power, and have yet to identify any convincingly validated modifiers of the BRCA1 and BRCA2 phenotype. To generate sufficient statistical power to identify modifier genes, the Consortium of Investigators of Modifiers of BRCA1 and BRCA2 (CIMBA) has been established. CIMBA contains about 30 affiliated groups who together have collected DNA and clinical data from approximately 10,000 BRCA1 and 5,000 BRCA2 mutation carriers. Initial efforts by CIMBA to identify modifiers of breast cancer risk for BRCA1 and BRCA2 mutation carriers have focused on validation of common genetic variants previously associated with risk in smaller studies of carriers or unselected breast cancers. Future studies will involve replication of findings from pathway-based and genome-wide association studies in both unselected and familial breast cancer. The identification of genetic modifiers of breast cancer risk for BRCA1 and BRCA2 mutation carriers will lead to an improved understanding of breast cancer and may prove useful for the determination of individualized risk of cancer amongst carriers. PMID- 17466084 TI - Myocardial injury associated with hyperinflation of the lung. PMID- 17466085 TI - Acute stroke: we have the treatments and we have the evidence--we need to use them. AB - Despite huge global burden, stroke disease has traditionally received little attention in the general medical press. We review a series of four acute stroke research articles published in a themed issue of the Lancet. Claiborne-Johnston and coworkers presented a scoring system to stratify risk of stroke following transient ischaemic attack. Chalela and colleagues demonstrated that magnetic resonance imaging is superior to computed tomography in detecting acute ischaemic stroke and that fears of missing intracranial haemorrhage are unfounded. The SITS MOST (Safe Implementation of Thrombolysis in Stroke - Monitoring Study) group reported positive experience of translating acute stroke thrombolysis trials into routine clinical practice in Europe, and the PROSIT (Project on Stroke Services in Italy) group studied acute effects of admission to a dedicated stroke unit. The message from all of these reports is that evidence-based, successful management of acute stroke is possible, and that investment in health infrastructure and changing mind sets of health practitioners to improve stroke care will deliver benefits. PMID- 17466086 TI - Sleep disturbances in spousal caregivers of individuals with Alzheimer's disease. AB - BACKGROUND: Although sleep problems are commonly reported among dementia caregivers, the nature and frequency of caregiver sleep disruptions, and their relationship to health status, has received little empirical attention to date. METHODS: The current study investigated the sleep situations of a sample of 60 spousal caregivers currently residing with a Alzheimer disease care recipient, including the frequency of nocturnal disruptions by the care recipient, and the reasons for these disruptions. In addition, exploratory correlations were computed between caregiver sleep variables and health outcomes. RESULTS: Some 63% of spousal caregivers reported sleep disruptions due to the nocturnal behavior of the recipients of their care. Poorer caregiver sleep quality was associated with higher frequency of nocturnal disruptions by the care recipient, the care recipient needing to use the bathroom, and wandering, higher caregiver depressive symptoms, and higher levels of caregiver role burden. The frequency of nocturnal disruptions was associated with poorer mental health status and a greater number of depressive symptoms. CONCLUSIONS: Results suggest that nocturnal disruptions by the care recipient may have adverse health consequences for spousal caregivers, and that further study of the determinants of caregiver sleep quality and health outcomes are warranted. PMID- 17466087 TI - Psychiatric disorders among the elderly on non-psychiatric wards in an African setting. AB - BACKGROUND: The elderly are vulnerable to illness and particularly to psychiatric illness. Many mentally ill elderly patients end up on non-psychiatric wards owing to somatization of their illnesses. Even for these patients, a psychiatric diagnosis may not be made. The literature on the elderly in Uganda is very scanty. This study aims to establish the prevalence and factors associated with psychiatric disorders among elderly patients admitted to non-psychiatric wards. METHODS: We carried out a descriptive cross-sectional study of 127 consenting elderly patients. They were administered a standardized questionnaire comprising the Self Reporting Questionnaire 25, the Mini-mental State Examination and the Structured Clinical Interview for the Diagnostic and Statistical Manual IV. Study variables included socio-demographic characteristics, physical illnesses, psychiatric disorders and the treatment given. RESULTS: The rate of psychiatric morbidity was 48%. The sex ratio was 1:1; however, women had a higher rate of psychiatric illness than men, 54.6% and 41.3% respectively. Being widowed or separated and having cancer were associated with SRQ>5, p=0.02 and p=0.04 respectively. Depressive disorders were the most common at 25.2% and were more common in women. Increasing age was associated with dementia (p<0.00). CONCLUSION: There is a high rate of psychiatric morbidity among the elderly in Uganda. Particular attention should be given to the psychological health of elderly people admitted to general hospitals. PMID- 17466088 TI - Congenital adrenal hyperplasia: focus on the molecular basis of 21-hydroxylase deficiency. AB - Congenital adrenal hyperplasia (CAH) is an autosomal recessive disorder caused by defects in one of several steroidogenic enzymes involved in the synthesis of cortisol from cholesterol in the adrenal glands. More than 90% of cases are caused by 21-hydroxylase deficiency, and the severity of the resulting clinical symptoms varies according to the level of 21-hydroxylase activity. 21-Hydroxylase deficiency is usually caused by mutations in the CYP21A2 gene, which is located on the RCCX module, a chromosomal region highly prone to genetic recombination events that can result in a wide variety of complex rearrangements, such as gene duplications, gross deletions and gene conversions of variable extensions. Molecular genotyping of CYP21A2 and the RCCX module has proved useful for a more accurate diagnosis of the disease, and prenatal diagnosis. This article summarises the clinical features of 21-hydroxylase deficiency, explains current understanding of the disease at the molecular level, and highlights recent developments, particularly in diagnosis. PMID- 17466089 TI - Long-term evaluation of extracorporeal shock wave lithotripsy in the treatment of salivary stones. AB - Extracorporeal shock wave lithotripsy is a rather new therapeutical method in the treatment of sialolithiasis. The objective was to evaluate retrospectively the results of the extracorporeal shock wave lithotripsy therapy performed with a Minilith SL 1 lithotripter on 167 out-patients with symptomatic stones (average size 5.94 mm) of the salivary glands over an observation period of seven years. A successful treatment with total stone disintegration was achieved in 51 (31 per cent) patients. In 92 (55 per cent) patients treatment was partially successful, with disappearance of the symptoms but a sonographically still identifiable stone. Treatment failure occurred in 24 (14 per cent) patients who then underwent surgery. The mean follow-up period was 35.6 months (minimum three, maximum 83), after which 83.2 per cent of the initially successfully treated patients were still free of symptoms.Therefore, extracorporeal shock wave lithotripsy, as a non invasive treatment alternative with few side effects, is an efficient technique for the therapy of sialolithiasis in selected patients. PMID- 17466090 TI - Anterior fracture dislocation of the odontoid peg in ankylosing spondylitis as a cause for rhinolalia clausa: a case study. AB - We present an unusual cause of rhinolalia clausa secondary to an oropharyngeal mass. A 69-year-old male presented to the otorhinolaryngology clinic with a one year history of a 'plummy' voice. He had a longstanding history of severe ankylosing spondylitis. Examination revealed an obvious hyponasal voice and a smooth hard mass in the midline of the posterior nasopharyngeal and oropharyngeal walls. Subsequent computed tomography scans and lateral plain neck X-ray showed a fracture dislocation of the odontoid peg, secondary to ankylosing spondylitis, which had eroded through the body of the C1 vertebra to lie anteriorly, resulting in the aforementioned impression into the pharyngeal mucosa. The radiological images, the role of the nasal airways in phonation and the causes of hyponasal speech are discussed. PMID- 17466091 TI - Digesta transit in different segments of the gastrointestinal tract of pigs as affected by insoluble fibre supplied by wheat bran. AB - Digestibility is the result of two competing processes: digestion and digesta transit. To develop or parameterise mechanistic models of digestion, both processes have to be quantified. The aim of this study was to determine the effect of insoluble dietary fibre on the transit in the gastrointestinal tract of pigs. Six barrows (33 kg initial body weight and fitted with two simple T cannulas at the proximal duodenum and distal ileum) were used in a double 3 x 3 Latin square design. Pigs were offered diets differing in total dietary fibre content (170, 220 and 270 g/kg DM) at 4 h intervals. A single meal marked with YbO2 and Cr-EDTA was used to determine the kinetics of markers concentrations of the solid and liquid phases, respectively. The mean retention time (MRT), calculated by the method of the moments, averaged 1, 4 and 38 h in the stomach, small intestine and large intestine, respectively. Increasing the insoluble fibre content in the diet had no effect on MRT in the stomach and decreased the MRT of both phases in the small intestine (P < 0.05). In the large intestine, increasing the insoluble fibre content decreased the MRT of the liquid phase (P = 0.02) and tended to decrease the MRT of the solid phase (P = 0.06). Transit of the solid phase in the large intestine was 4-8 h slower than transit of the liquid phase. Analysis of marker excretion curves indicated that the small and large intestine should be represented mathematically to have both a tubular (propulsion) and compartmental (mixing) structure. PMID- 17466092 TI - Iron deficiency and NRAMP1 polymorphisms (INT4, D543N and 3'UTR) do not contribute to severity of anaemia in tuberculosis in the Indonesian population. AB - Fe-deficiency anaemia is the most common cause of anaemia in developing countries. In these settings, many chronic infections, including tuberculosis (TB), are highly prevalent. Fe is an essential nutrient for both host and mycobacteria that play a pivotal role in host immunity and mycobacterial growth. A case-control study was performed in a TB-endemic region in Jakarta, Indonesia, among 378 pulmonary TB patients and 436 healthy controls from the same neighbourhood with the same socio-economic status. In a number of these subjects the Fe status could be explored. The distribution of three polymorphisms in the natural resistance-associated macrophage protein gene (NRAMP1) including INT4, D543N and 3'UTR was examined for a possible association with susceptibility to TB. Anaemia (corrected for sex) was present in 63.2 % of active TB compared with 6.8 % of controls, with female patients more often affected. Anaemia was more pronounced in advanced TB as diagnosed by chest radiography. Lower Hb concentrations in TB patients were accompanied by lower plasma Fe concentrations, lower Fe-binding capacity and higher plasma ferritin. After successful TB therapy, Fe parameters improved towards control values and Hb levels normalised, even without Fe supplementation. NRAMP1 gene polymorphisms were not associated with TB susceptibility, TB severity or anaemia. In conclusion, most active TB patients had anaemia, which was probably due to inflammation and not to Fe deficiency since TB treatment without Fe supplementation was sufficient to restore Hb concentration. PMID- 17466093 TI - Dietary taurine reduces retinal damage produced by photochemical stress via antioxidant and anti-apoptotic mechanisms in Sprague-Dawley rats. AB - Taurine has been shown to be tissue protective in many models of oxidant-induced injury. However, its protective role against retinal damage induced by photochemical stress is less well known. The purpose of the present study was to investigate whether dietary taurine reduced retinal photochemical damage in Sprague-Dawley rats and to further explore the underlying molecular mechanisms of this action. Twenty rats fed AIN-93 formulation and maintained in the dark for 48 h were used as controls (n 20). Another forty rats were randomly divided into two groups and then treated with (n 20) or without 4 % taurine (n 20) for 15 d respectively. After treatment, these two groups were exposed to fluorescent light (3000 +/- 200 lux and 25 degrees C), and the protective effects of dietary taurine were then evaluated. The present results showed that dietary taurine effectively prevented retinal photochemical damage as assessed by changes of morphology. Also, the supplementation caused an increase of taurine in the retina, a decrease of malondialdehyde (P < 0.01), and elevation of superoxide dismutase (P < 0.01) and glutathione peroxidase activities in the retina (P < 0.01). Moreover, dietary taurine inhibited activator protein-1 (AP-1) (c-fos/c jun subunits) expression (P < 0.05), up regulated NF-kappaB (p65) expression (P < 0.05), and decreased caspase-1 expression (P < 0.05) so as to reduce the apoptosis of photoreceptors in the retina (P < 0.05). These results suggest that dietary taurine reduced retinal damage produced by photochemical stress via antioxidant and anti-AP-1-NF-kappaB-caspase-1 apoptotic mechanisms in rats. PMID- 17466094 TI - Effects of partial substitution of dietary fish oil with blends of vegetable oils, on blood leucocyte fatty acid compositions, immune function and histology in European sea bass (Dicentrarchus labrax L). AB - Within a decade or so insufficient fish oil (FO) will be available to meet the requirements for aquaculture growth. Consequently, alternative sources are being investigated to reduce reliance on wild fish as a source of FO. Vegetable oils (VO) are a feasible alternative to FO. However, it is important to establish that alternative dietary lipids are not only supplied in the correct quantities and balance for optimal growth, but can maintain immune function and prevent infection, since it is known that the nutritional state of the fish can influence their immune function and disease resistance. A way of maintaining immune function, while replacing dietary FO, is by using a blend of VO rather than a single oil. In this study, juvenile European sea bass (Dicentrarchus labrax) were fed diets with a 60 % substitution of FO with a blend of rapeseed, linseed and palm oils. Two oil blends were used to achieve a fatty acid composition similar to FO, in terms of energy content, and provide a similar balance of SFA, MUFA and PUFA. Fish were fed the diets for 64 weeks, after which time growth and fatty acid compositions of liver and blood leucocytes were monitored. The impact of the dietary blends on selected innate immune responses and histopathology were also assessed, together with levels of plasma prostaglandin E2. The results suggest that potential exists for replacing FO with a VO blend in farmed sea bass feeds without compromising growth, non-specific immune function or histology. PMID- 17466095 TI - The intake of flavonoids and carotid atherosclerosis: the Kuopio Ischaemic Heart Disease Risk Factor Study. AB - The role of flavonoids in CVD is still unclear. In this cross-sectional study we assessed the relation between the intakes of twenty-six flavonoids from five subclasses: flavonols, flavones, flavanones, flavan-3-ols and anthocyanidins, and the mean common carotid artery intima-media thickness (CCA-IMT). The study population consisted of 1380 middle-aged eastern Finnish men for whom the mean CCA-IMT examinations were carried out as a part of the prospective population based Kuopio Ischaemic Heart Disease Risk Factor Study (KIHD). The mean intake of flavonoids was 128.5 (sd 206.7) mg/d and the mean CCA-IMT was 0.78 (sd 0.17) mm. In the lowest quartile of total flavonoid intake the non-adjusted mean CCA-IMT was 0.79 (sd 0.19) mm, while the mean CCA-IMT was 0.76 (sd 0.15) in the highest quartile (P < 0.001). After adjustment for age, variables related to CCA-IMT measurement, history of atherosclerosis, smoking, BMI, diabetes, systolic blood pressure, serum HDL- and LDL-cholesterol, VO2 max, and intakes of alcohol, SFA, folate, vitamins C and E, the total flavonoid intake was inversely associated with the mean CCA-IMT (P = 0.018). Out of different flavonoid subclasses, flavan 3-ols were inversely associated with CCA-IMT (P = 0.025) after statistical adjustment. There was a trend for an inverse association between intake of flavonols and mean CCA-IMT (P = 0.055). We conclude that high intake of flavonoids is associated with decreased carotid atherosclerosis in middle-aged Finnish men. PMID- 17466096 TI - Effects of bioactive peptides isoleucine-proline-proline (IPP), valine-proline proline (VPP) and leucine-lysine-proline (LKP) on gene expression of osteoblasts differentiated from human mesenchymal stem cells. AB - Food-derived bioactive peptides are reported to express a variety of functions in vivo. We studied the in vitro effect of three bioactive tripeptides, isoleucine proline-proline (IPP), valine-proline-proline (VPP) and leucine-lysine-proline (LKP), on osteoblast proliferation and gene expression. We used UMR-106 osteosarcoma cells, human marrow-derived mesenchymal stem cells (hMSC) and osteoblasts differentiated from hMSC. Treatment with 50 mum-IPP increased UMR-106 cell and hMSC proliferation. The gene expression of hMSC-differentiated osteoblasts was analysed by the microarray method. Microarray analysis revealed that IPP up-regulated 270 genes and down-regulated 100 genes. VPP and LKP, by contrast, had a very modest influence on osteoblast gene expression. Real-time PCR confirmed that IPP up-regulated PTHrP, BMP-5 and CREB-5 and down-regulated VDR and caspase-8. IPP possesses potential to increase osteoblast proliferation, differentiation and signalling. Agents that increase the number and function of osteoblasts could improve bone mass and structure, and decrease fracture risk. PMID- 17466097 TI - A comparison of the programme coverage of two therapeutic feeding interventions implemented in neighbouring districts of Malawi. AB - OBJECTIVE: To compare therapeutic feeding programme coverage for severely malnourished children achieved by a community-based therapeutic care (CTC) programme and a therapeutic feeding centre (TFC) programme operating in neighbouring districts in Malawi. DESIGN: Two surveys were implemented simultaneously one in each of the two programme areas. Each survey used a stratified design with strata defined using the centric systematic area sample method. Thirty 100 km2 quadrats were sampled. The community or communities located closest to the centre of each quadrat were sampled using a case-finding approach. Cases were defined as children aged under 5 years with 3600 publications cited in PubMed. Although originally described as a mediator of lipoprotein metabolism, the lipoprotein-independent functions of apoE are being increasingly recognised, with limited data available on the potential impact of genotype on these metabolic processes. Furthermore, although meta-analyses suggest that apoE4 carriers may have a 40-50% increased CVD risk, the associations reported in individual studies are highly heterogeneous and it is recognised that environmental factors such as smoking status and dietary fat composition influence genotype-phenotype associations. However, information is often derived from observational studies or small intervention trials in which retrospective genotyping of the cohort results in small group sizes in the rarer E2 and E4 subgroups. Either larger well-standardised intervention trials or smaller trials with prospective recruitment according to apoE genotype are needed to fully establish the impact of diet on genotype-CVD associations and to establish the potential of dietary strategies such as reduced total fat, saturated fat, or increased antioxidant intakes to counteract the increased CVD burden in apoE4 carriers. PMID- 17466102 TI - Developmental programming of energy balance and the metabolic syndrome. AB - The increasing prevalence of the metabolic syndrome in numerous populations throughout the world is currently of major concern, and presents a huge global health problem. The link between low birth weight and the subsequent development of obesity, disrupted glucose homeostasis and hypertension is now well established, and there is extensive evidence supporting these associations in both epidemiological and experimental studies. Alterations in the secretion of, and responses to, the circulating hormones insulin and leptin are likely candidates in terms of disease development. The aim of current research is to define how the central and peripheral pathways in which these signals exert their effects may be disrupted following poor early growth, and how this disruption contributes to the development of metabolic disease. The present review aims to outline the existing evidence whereby alterations in early growth may programme an individual to be at increased risk of the metabolic syndrome. The development of central appetite and expenditure circuits and of peripheral metabolic tissues, are likely to play a key role in the long-term regulation of energy balance. PMID- 17466103 TI - Phytochemicals and cancer. AB - Epidemiological studies showing a protective effect of diets rich in fruits and vegetables against cancer have focused attention on the possibility that biologically-active plant secondary metabolites exert anti-carcinogenic activity. This huge group of compounds, now collectively termed 'phytochemicals', provides much of the flavour and colour of edible plants and the beverages derived from them. Many of these compounds also exert anti-carcinogenic effects in animal models of cancer, and much progress has been made in defining their many biological activities at the molecular level. Such mechanisms include the detoxification and enhanced excretion of carcinogens, the suppression of inflammatory processes such as cyclooxygenase-2 expression, inhibition of mitosis and the induction of apoptosis at various stages in the progression and promotion of cancer. However, much of the research on phytochemicals has been conducted in vitro, with little regard to the bioavailability and metabolism of the compounds studied. Many phytochemicals present in plant foods are poorly absorbed by human subjects, and this fraction usually undergoes metabolism and rapid excretion. Some compounds that do exert anti-carcinogenic effects at realistic doses may contribute to the putative benefits of plant foods such as berries, brassica vegetables and tea, but further research with human subjects is required to fully confirm and quantify such benefits. Chemoprevention using pharmacological doses of isolated compounds, or the development of 'customised' vegetables, may prove valuable but such strategies require a full risk-benefit analysis based on a thorough understanding of the long-term biological effects of what are often surprisingly active compounds. PMID- 17466104 TI - Life-course approaches to inequalities in adult chronic disease risk. AB - A life-course approach to chronic-disease epidemiology uses a multidisciplinary framework to understand the importance of time and timing in associations between exposures and outcomes at the individual and population levels. Such an approach to chronic diseases is enriched by specification of the particular manner in which timing in relation to physical growth, reproduction, infection, social mobility, behavioural transitions etc. can influence various adult chronic diseases in different ways, and more ambitiously by how these temporal processes are interconnected and manifested in health inequalities within a population and in population-level disease trends. The paper will discuss some historical background to life-course epidemiology and theoretical models of life-course processes, and will review some of the empirical evidence linking life-course processes to CHD, haemorrhagic stroke, stomach cancer and other chronic diseases in adulthood. It will also underscore that a life-course approach offers a way to conceptualize how underlying socio-environmental determinants of health, experienced at different life-course stages, can differentially influence the development of chronic diseases, as mediated through proximal specific biological processes. PMID- 17466105 TI - Differential immunomodulation with long-chain n-3 PUFA in health and chronic disease. AB - The balance of intake of n-6 and n-3 PUFA, and consequently their relative incorporation into immune cells, is important in determining the development and severity of immune and inflammatory responses. Some disorders characterised by exaggerated inflammation and excessive formation of inflammatory markers have become among the most important causes of death and disability in man in modern societies. The recognition that long-chain n-3 PUFA have the potential to inhibit (excessive) inflammatory responses has led to a large number of clinical investigations with these fatty acids in inflammatory conditions as well as in healthy subjects. The present review explores the presence of dose-related effects of long-chain n-3 PUFA supplementation on immune markers and differences between healthy subjects and those with inflammatory conditions, because of the important implications for the transfer of information gained from studies with healthy subjects to patient populations, e.g. for establishing dose levels for specific applications. The effects of long-chain n-3 PUFA supplementation on ex vivo lymphocyte proliferation and cytokine production by lymphocytes and monocytes in healthy subjects have been studied in twenty-seven, twenty-five and forty-six treatment cohorts respectively, at intake levels ranging from 0.2 g EPA+DHA/d to 7.0 g EPA+DHA/d. Most studies, particularly those with the highest quality study design, have found no effects on these immune markers. Significant effects on lymphocyte proliferation are decreased responses in seven of eight cohorts, particularly in older subjects. The direction of the significant changes in cytokine production by lymphocytes is inconsistent and only found at supplementation levels > or =2.0 g EPA+DHA/d. Significant changes in inflammatory cytokine production by monocytes are decreases in their production in all instances. Overall, these studies fail to reveal strong dose-response effects of EPA+DHA on the outcomes measured and suggest that healthy subjects are relatively insensitive to immunomodulation with long-chain n-3 PUFA, even at intake levels that substantially raise their concentrations in phospholipids of immune cells. In patients with inflammatory conditions cytokine concentrations or production are influenced by EPA+DHA supplementation in a relatively large number of studies. Some of these studies suggest that local effects at the site of inflammation might be more pronounced than systemic effects and disease-related markers are more sensitive to the immunomodulatory effects, indicating that the presence of inflamed tissue or 'sensitised' immune cells in inflammatory disorders might increase sensitivity to the immunomodulatory effects of long chain n-3 PUFA. In a substantial number of these studies clinical benefits related to the inflammatory state of the condition have been observed in the absence of significant effects on immune markers of inflammation. This finding suggests that condition-specific clinical end points might be more sensitive markers of modulation by EPA+DHA than cytokines. In general, the direction of immunomodulation in healthy subjects (if any) and in inflammatory conditions is the same, which indicates that studies in healthy subjects are a useful tool to describe the general principles of immunomodulation by n-3 PUFA. However, the extent of the effect might be very different in inflammatory conditions, indicating that studies in healthy subjects are not particularly suitable for establishing dose levels for specific applications in inflammatory conditions. The reviewed studies provide no indications that the immunomodulatory effects of long-chain n-3 PUFA impair immune function or infectious disease resistance. In contrast, in some conditions the immunomodulatory effects of EPA+DHA might improve immune function. PMID- 17466106 TI - Nutritional management of gut health in pigs around weaning. AB - Early weaning of piglets is often accompanied by a severe growth check and diarrhoea. It is well established that this process is multi-factorial and that post-weaning anorexia and undernutrition are major aetiological factors. Gastrointestinal disturbances include alterations in small intestine architecture and enzyme activities. Recent data indicate transiently-increased mucosal permeability, disturbed absorptive-secretory electrolyte balance and altered local inflammatory cytokine patterns after weaning. These responses appear to operate according to two distinct temporal patterns, an acute response followed by a long-lasting adaptation response. Pigs coexist with a diverse and dense commensal microbiota in their gastrointestinal tract. Most of these microbes are beneficial, providing necessary nutrients or protection against harmful pathogens for the host. The microbial colonisation of the porcine intestine begins at birth and follows a rapid succession during the neonatal and weaning period. Following the withdrawal of sow's milk the young piglets are highly susceptible to enteric diseases partly as a result of the altered balance between developing beneficial microbiota and the establishment of intestinal bacterial pathogens. The intestinal immune system of the newborn piglet is poorly developed at birth and undergoes a rapid period of expansion and specialisation that is not achieved before early (commercial) weaning. Here, new insights on the interactions between feed components, the commensal microbiota and the physiology and immunology of the host gastrointestinal tract are highlighted, and some novel dietary strategies are outlined that are focused on improving gut health. Prebiotics and probiotics are clear nutritional options, while convincing evidence is still lacking for other bioactive substances of vegetable origin. PMID- 17466107 TI - Professional regulation of nutritionists: where are we now? AB - Waterlow's (1981) Sixth Boyd Orr Lecture on a 'crisis of identity for nutrition' stimulated the Nutrition Society's drive to professionalisation. Twenty-five years on, the Society begins a new stage; first, towards an independent voluntary regulator, and then towards statutory regulation. It is timely to reflect on progress and identify the remaining challenges. The Society has made impressive progress as a voluntary regulator since 1991 when the Institute of Biology opened a register in cooperation with the Institute of Food Science and Technology and the Nutrition Society; the present register is 2.75-fold larger. The Society has specialist standards for course accreditation that enable graduates to apply for direct entry to the register, having met standards of competency in nutrition or public health nutrition. A code of ethics and a statement of professional conduct underpin a functioning system for oversight and governance that protects the public, the hallmark of all professions. Registered nutritionists lay easy claim to a unique science basis for their profession. A scheme for continuing professional development (CPD) started in July 2006, 1 year before a sample audit starts to show the link between CPD and re-registration. The scheme will be piloted in the first year. The critical challenge is the issue of identity. Waterlow (1981) stated that professional registration must lead explicitly and formally to a specific vocation, an occupation that provides services that society requires and one that contributes to the well-being and health of all. The present time may be the last and best chance for nutritionists, as nutrition has a higher priority for government in the UK than ever before. The Society has begun to help in strategic public health workforce planning and development; new and still plastic, it is the ideal locus from which a discipline and a profession can emerge. The CPD scheme will work if it helps nutritionists meet their own needs; more mutual cooperation and consensus about real world standards of performance are needed. Nutritionists need to show how they actually contribute to national health and/or wealth. Then, sustained resources can be advocated for and the support of the voting public and legislators secured, without which it will not be possible to get the legal protection that is desirable for the profession. PMID- 17466108 TI - The psychology of food craving. AB - Cravings are hedonic responses to food, characterised by their intensity and their specificity. Food cravings are extremely common, reported by the majority of young adults. They are closely associated with liking but not synonymous with increased intake. Structured interviews and prospective incident accounts of food cravings have succeeded in revealing a richness of information about their character, their antecedents and their consequences. In addition, laboratory investigations are adding to what is being learned from field and clinical studies. Taking dieting as an example of an assumed influence on food craving, the outcomes of cross-sectional studies are mixed and unconvincing. Prospective and experimental research shows a clearer relationship. Dieting or restrained eating generally increase the likelihood of food craving while fasting makes craving, like hunger, diminish. Attempted restriction or deprivation of a particular food is associated with an increase in craving for the unavailable food. This relationship suggests a variety of underlying cognitive, conditioning and emotional processes, of which ironic cognitive processes, conditioned cue reactivity and dysphoric mood are prominent. Food cravings may also be self attributions, accounting for why a highly-palatable but self-restricted food is (over-)consumed. Overall, the popularised account of cravings as elicited by specific nutritional need is having to give way to a more subtle and complex appreciation of human eating behaviour. PMID- 17466109 TI - CVD risk in South Asians: the importance of defining adiposity and influence of dietary polyunsaturated fat. AB - The prevalence of the metabolic syndrome (MetS), CVD and type 2 diabetes (T2D) is known to be higher in populations from the Indian subcontinent compared with the general UK population. While identification of this increased risk is crucial to allow for effective treatment, there is controversy over the applicability of diagnostic criteria, and particularly measures of adiposity in ethnic minorities. Diagnostic cut-offs for BMI and waist circumference have been largely derived from predominantly white Caucasian populations and, therefore, have been inappropriate and not transferable to Asian groups. Many Asian populations, particularly South Asians, have a higher total and central adiposity for a similar body weight compared with matched Caucasians and greater CVD risk associated with a lower BMI. Although the causes of CVD and T2D are multi factorial, diet is thought to make a substantial contribution to the development of these diseases. Low dietary intakes and tissue levels of long-chain (LC) n-3 PUFA in South Asian populations have been linked to high-risk abnormalities in the MetS. Conversely, increasing the dietary intake of LC n-3 PUFA in South Asians has proved an effective strategy for correcting such abnormalities as dyslipidaemia in the MetS. Appropriate diagnostic criteria that include a modified definition of adiposity must be in place to facilitate the early detection and thus targeted treatment of increased risk in ethnic minorities. PMID- 17466110 TI - Internet-based cognitive behavioural therapy for subthreshold depression in people over 50 years old: a randomized controlled clinical trial. AB - BACKGROUND: Subthreshold depression is a highly prevalent condition and a risk factor for developing a major depressive episode. Internet-based cognitive behaviour therapy may be a promising approach for the treatment of subthreshold depression. The current study had two aims: (1) to determine whether an internet based cognitive behaviour therapy intervention and a group cognitive behaviour therapy intervention are more effective than a waiting-list control group; and (2) to determine whether the effect of the internet-based cognitive behaviour therapy differs from the group cognitive behaviour therapy intervention. METHOD: A total of 191 women and 110 men with subthreshold depression were randomized into internet-based treatment, group cognitive behaviour therapy (Lewinsohn's Coping With Depression course), or a waiting-list control condition. The main outcome measure was treatment response after 10 weeks, defined as the difference in pre- and post-treatment scores on the Beck Depression Inventory (BDI). Missing data, a major limitation of this study, were imputed using the Multiple Imputation (MI) procedure Data Augmentation. RESULTS: In the waiting-list control group, we found a pre- to post-improvement effect size of 0.45, which was 0.65 in the group cognitive behaviour therapy condition and 1.00 within the internet based treatment condition. Helmert contrasts showed a significant difference between the waiting-list condition and the two treatment conditions (p=0.04) and no significant difference between both treatment conditions (p=0.62). CONCLUSIONS: An internet-based intervention may be at least as effective as a commonly used group cognitive behaviour therapy intervention for subthreshold depression in people over 50 years of age. PMID- 17466111 TI - Exploring the inter-relationship of smoking age-at-onset, cigarette consumption and smoking persistence: genes or environment? AB - BACKGROUND: We investigated the genetic and environmental contributions to covariation between smoking age-at-onset, cigarette consumption and smoking persistence. METHOD: Multivariate biometrical modelling methods were applied to questionnaire data from Australian twins and their siblings (14 472 individuals from 6247 families). The contributions of genetic and environmental factors to covariation between the three traits were estimated, allowing for sex differences in both trait prevalence and the magnitude of genetic and environmental effects. RESULTS: All traits were moderately heritable in males and females (estimates between 0.40 and 0.62), but there were sex differences in the extent to which additive genetic influences were shared across traits. Twin-specific environmental factors accounted for a substantial proportion of the variance in smoking age-at-onset in females (0.19) and males (0.12), but had little influence (<0.08) on other traits. Unique environmental factors were estimated to have a moderate influence on smoking age-at-onset (0.17 for females, 0.19 for males), but a stronger influence on other traits (between 0.39 and 0.49). CONCLUSIONS: These results provide some insight into observed sex differences in smoking behaviour, and suggest that searching for pleiotropic genes may prove fruitful. However, further work on phenotypic definitions of smoking behaviour, particularly persistence, is warranted. PMID- 17466112 TI - Hygienic quality of ewes' milk cheeses manufactured with artisan-produced lamb rennet pastes. AB - The use of artisan-produced lamb rennet pastes, but not any of the other commercial animal rennets, imparts a characteristic flavour to the cheese, so most Mediterranean ewes' milk cheeses are coagulated with this kind of rennet paste. In contrast to the advantages of using lamb or kid rennet pastes from the sensory point of view, questions are still raised as to their hygienic quality. The goal was to examine the microbiological and enzymic quality of lamb rennet pastes prepared by cheese manufacturers for their own use, and evaluate the hygienic quality of raw sheeps' milk cheeses made with them, using Idiazabal cheese as a model. Lamb rennet pastes prepared by artisan cheese makers from the Basque region of Spain (27), and Italy (8) were evaluated. For cheese making experiments 5 different lamb rennet pastes were selected among the 27 samples from the Basque Country region of Spain. Microbiological analyses were carried out on samples from rennet pastes, rennet extracts, milks and cheeses during ripening. Enzymic activities studied in rennet paste were: total coagulating strength and lipase. Analysis of variance and Student's t-tests was performed. The results show that the artisan-produced rennet pastes contain high levels of a variety of microorganisms. After 60 ripening days, which is the minimum ripening period required for Idiazabal cheese prior to its commercialization, no Eschericia coli, Clostridium, Salmonella spp. or Listeria monocytogenes were detected, and levels for the rest of the microorganisms were below the limits of the European legislative standards for cheese manufactured with raw milk. We can conclude that the use of artisan-produced lamb rennet pastes of questionable hygienic quality for the manufacture of raw milk hard cheeses yields products of good hygienic quality. PMID- 17466113 TI - Development of a monoclonal antibody-based immunoassay for specific quantification of bovine milk alkaline phosphatase. AB - The detection of alkaline phosphatase (ALP) activity is used as a legal test to determine whether milk has been adequately pasteurized or recontaminated with raw milk. However, a wide variety of microorganisms produce both heat labile and heat stable ALPs which cannot be differentiated from the milk ALP by current enzymatic methods. Monoclonal antibodies specific of the bovine milk ALP were obtained in mice from a raw bovine milk ALP preparation. Coated in microtitre plates, these antibodies specifically capture the bovine milk ALP from dairy products. After washing, the enzymatic activity of the captured ALP is revealed by adding p nitrophenyl-phosphate as a substrate. This simple immunoassay does not react with ALPs of intestinal or bacterial origin and, once optimized, was found to be the first immunoassay suitable to detect raw milk in boiled milk down to a 0.02% dilution. Moreover, in contrast with competitive indirect ELISA formats, the capture immunoassay does not require purified ALP. PMID- 17466114 TI - Characterization of Fiore Sardo cheese manufactured with the addition of autochthonous cultures. AB - This work evaluated the effect of adjunct autochthonous cultures on the chemical, microbiological and sensory characteristics of Fiore Sardo cheese during ripening. A total of twelve batches of cheeses were manufactured according to the technical Disciplinary of Fiore Sardo cheese, with and without different combinations of autochthonous strains isolated from the native microflora of artisanal Fiore Sardo. There were no significant differences in the cheese compositional parameters between experimental and control cheeses, but the addition of cultures led to a statistically significant decrease in pH values in experimental cheeses. The evolution of total mesophilic bacteria, total coliforms and lactic acid bacteria were significantly influenced by the addition of autochthonous cultures in most of the experimental cheeses. As for sensory characteristics, all the experimental cheeses reported significantly higher scores especially for shape, texture, interior openings, taste and aftertaste. This study demonstrated the beneficial effect of the addition of selected autochthonous cultures in accelerating the disappearance of undesirable flora and improving the typical sensory characteristics of the cheese, and confirmed the importance of ewes' milk as a source of technologically interesting strains that could be used to ensure a higher quality of artisanal cheese productions. PMID- 17466115 TI - In vitro reconstitution of antimicrobial pathogen activity by expressed recombinant bovine lactoferrin N-terminal peptide in Escherichia coli. AB - Recombinant bovine lactoferrin N-terminal polypeptide (rbLF-N) Escherichia coli expression system was constructed and the rbLF-N antimicrobial activity was displayed by enzymatic proteolysis in this study. A 162 bp 5'-terminal fragment of bovine lactoferrin (bLF) gene from bovine liver gDNA was amplified by PCR. The DNA fragment containing exon-2 of the bLF gene was cloned into the expression vector pGEX-4T1 and the glutathione-S-transferase-rbLF-N (GST-rbLF-N) fusion protein was obtained by over-expression in Esch. coli BL21(DE3). After thrombin/pepsin digestion, the rbLF-N was released from the fusion protein. The recombinant peptide was separated and identified by SDS-PAGE, HPLC and LC-MS/MS analysis. A very strong anti-food-born microbial pathogen activity of the rbLF-N peptides was displayed through bio- and kinetic-assays in vitro. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of the rbLF-N peptide for bacterial pathogens Staphylococcus aureus, Streptococcus mutans, Esch. coli and Klebsiella pneumoniae were 11.7, 11.7, 11.7, 23.4 microg and 23.4, 11.7, 11.7, 46.4 microg, respectively. This study created a new route for exploring lactoferrin peptide application in food science. PMID- 17466116 TI - Antibiotic survey of Lactococcus lactis strains to six antibiotics by Etest, and establishment of new susceptibility-resistance cut-off values. AB - In order to establish cut-off values for Lactococcus lactis to six antibiotics to distinguish susceptible and intrinsically resistant strains from those having acquired resistances, the minimum inhibitory concentration (MIC) of tetracycline, erythromycin, clindamycin, streptomycin, chloramphenicol and vancomycin was determined in 93 different Lc. lactis strains using the Etest. These bacterial strains were originally isolated from dairy and animal sources in widely separated geographical locations. Cut-offs were defined on the basis of the distribution of the MICs frequency of the studied antibiotics, which in the absence of acquired determinants should approach to a normal statistical distribution. In general, the new cut-off values proposed in this study are higher than previously defined (European Commission, 2005. The EFSA Journal 223, 1-12). Based on these new values, all the strains tested were susceptible to erythromycin, chloramphenicol and vancomycin, and 79 susceptible to all six antibiotics. However, 11 strains (around 12%) were considered resistant to tetracycline (six of which had been identified after screening of a large collection of lactococci strains for tetracycline resistance) and five (5.4%) resistant to streptomycin. Of these, two fish isolates proved to be resistance to both tetracycline and streptomycin. From the tetracycline resistant strains, tet(M) and mosaic tet(L/S) genes were amplified by PCR, demonstrating they harboured acquired antibiotic resistance determinants. PMID- 17466117 TI - Influence of pressure release rate and protein concentration on the formation of pressure-induced casein structures. AB - The formation of pressure-induced casein structures (600 MPa for 30 min at 30 degrees C) was investigated for different pressure release rates (20 to 600 MPa min-1) and casein contents (1 to 15 g/100 ml). Structures from liquid (sol) to solid (gel) were observed. The higher the protein content and the pressure release rate, the higher was the dynamic viscosity. A firm gel was built up at a casein content of 7 g/100 ml for a pressure release rate of 600 MPa min-1, while lower release rates resulted in less firm gels (200 MPa min-1) or liquid structures (20 MPa min-1). In a 5 g/100 ml casein solution and at a pressure release rate of 600 MPa min-1, casein aggregates were generated which were built from smaller casein particles with a larger hydrodynamic diameter and higher voluminosity than in the untreated solution. After a slow release rate casein micelles had a smaller hydrodynamic diameter and a lower voluminosity, but were similar in shape and diameter as compared with the micelles in solution before high pressure treatment. PMID- 17466118 TI - HPLC quantification of biogenic amines in cheeses: correlation with PCR-detection of tyramine-producing microorganisms. AB - The consumption of food and beverages containing high amounts of biogenic amines (BA) can have toxicological effects. BA found in foods and beverages are synthesized by the microbial decarboxylation of certain amino acids. This paper reports the concentrations of BAs in a number of commercial cheeses, as determined by HPLC. The cheeses studied were made from raw and pasteurized milk of different origin, and were subjected to different ripening periods. BA concentrations were lower in short ripening period than in long ripening period cheeses, and higher in cheeses made from raw milk than in those made from pasteurized milk. The highest BA concentrations were recorded in blue cheeses made from raw milk. Tyramine was the most commonly recorded and abundant BA. The presence of tyramine-producing bacteria was determined by PCR, and a good correlation obtained between the results of this method and tyramine detection by HPLC. These methods could be used to complement one another in the detection and quantification of tyramine in cheese prevention of tyramine accumulation in cheese. PMID- 17466119 TI - Inactivation-denaturation kinetics of bovine milk alkaline phosphatase during mild heating as determined by using a monoclonal antibody-based immunoassay. AB - A monoclonal antibody based capture immunoassay has been recently developed for the specific quantitation of bovine milk alkaline phosphatase (ALP) without interference by contaminating microbial or fungal ALPs (Geneix et al. 2007). This immunoassay was used to study the kinetics of ALP heat denaturation in bovine milk over a range 50-60 degrees C for 5 to 60 min using a colorimetric quantification of the enzyme activity as a reference test. A denaturation midpoint was obtained at 56 degrees C for a 30 min heating. Thermal inactivation was found to follow first order kinetics and is characterized by z value of 6.7 deg C (D60 degrees C=24.6 min) and 6.8 (D60 degrees C=23.0 min) for respectively immunoassay and colorimetric assay. The high values of enthalpy of activation and the positive values of the entropy of activation and free energy of activation indicate that during denaturation ALP underwent a large change in conformation. The results of the immunoassay were highly correlated (r=0.994) with those obtained by the colorimetric assay. A similar high correlation (r=0.998) was obtained when industrially thermized milks (62-67 degrees C for 20-90 s) were analysed by both techniques. These results indicated that 1) thermally induced epitopic structural changes recognized by the capture monoclonal antibody are concomitant with or occur after the loss of enzymatic activity and 2) quantification of ALP by the specific immunoassay is appropriate for determining mild time/temperature treatment of milk and for the control of milk pasteurization. PMID- 17466120 TI - Effects of intravenous triacylglycerol emulsions on lymphocyte responses to mitogens in fasted dairy cows undergoing intense lipomobilization. AB - The objective of the study was to assess the effects of intravenous infusion of triacylglycerol (TAG) emulsions derived from different lipid sources on responses to mitogens of peripheral blood mononuclear cells (PBMC) isolated from fasted dairy cows. Six multiparous, non-pregnant, non-lactating Holstein cows were used in a replicated 3x3 Latin Square design. For 4 d, cows were fasted and infused intravenously with a 20% TAG emulsions derived from tallow (TA), linseed oil (LO) or fish oil (FO). Fasting was employed to induce energy deficit and lipid mobilization. Emulsions were administered for 20 to 30 min every 4 h throughout the 4 d fast at a rate of 0.54 g TAG/kg BW/d. Blood samples were taken before the first infusion, and then every 24 h during the fast. Blood was utilized to assess DNA synthesis, IgM and interferon-gamma (IFN-gamma) secretion by PBMC stimulated with mitogens. In TA infused cows there was a decline of PBMC ability to respond to mitogens, which was significant 48 h after initiation of the infusion period for DNA synthesis and IFN-gamma secretion. In LO or FO infused cows, PBMC responses to mitogens were not altered during the infusion period, and in some cases PBMC responses to mitogen was improved at 72 and 96 h after initiation of treatments. Effects of TAG infusion on PBMC responses to mitogens depended on the lipid source suggesting that LO or FO can attenuate the negative effects of fasting on immune functions. PMID- 17466121 TI - Effect of simulated gastrointestinal digestion on the antihypertensive properties of synthetic beta-lactoglobulin peptide sequences. AB - In this study, the antihypertensive activity in spontaneously hypertensive rats of two peptides isolated from beta-lactoglobulin hydrolysates with thermolysin was evaluated. These peptides, with sequences LLF [beta-lg f(103-105)] and LQKW [beta-lg f(58-61)], showed potent in vitro ACE-inhibitory activity. Two hours after administration, both sequences caused a clear and significant decrease in the blood pressure of these rats. The impact of a simulated gastrointestinal digestion on ACE-inhibitory and antihypertensive activities of these peptides was also studied. The results showed that both fragments were susceptible to proteolytic degradation after incubation with pepsin and Corolase PP. In addition, their in vitro ACE-inhibitory activity decreased after the simulated digestion. It is likely that fragment LQK was the active end product of the gastrointestinal digestion of peptide LQKW. The fragment LL, observed after digestion of peptide LLF, probably exert its antihypertensive effect through a mechanism of action different than ACE-inhibition. PMID- 17466122 TI - Production from dairy cows of semi-industrial quantities of milk-protein concentrate (MPC) containing efficacious anti-Candida albicans IgA antibodies. AB - Bovine milk antibodies directed against human pathogenic organisms have potential as prophylactic or therapeutic treatments of disorders affecting mucosal surfaces. The cow, however, does not naturally secrete high levels of IgA in milk, the predominant immunoglobulin of the mucosal immune system. We have patented an immunisation protocol that results in increased production of IgA in ruminant milk and in this study established that our protocol can be used on a scale sufficient to produce semi-industrial quantities of milk for processing. Cows were immunised with a common pathogenic yeast, Candida albicans and responded with high levels of antigen-specific IgA antibodies in their milk. The spray-dried milk-protein concentrate (85% protein) powder was shown to reduce adherence of Cand. albicans cells in in vitro adherence assays, demonstrating an ability to retain efficacy through the processing. These results suggest that this milk product may be of therapeutic value if the reduction in Cand. albicans adhesion observed in vitro translates to reduced colonisation in vivo. PMID- 17466123 TI - Epinephrine in digital blocks: refuting the refutation of dogma. PMID- 17466124 TI - Chest pain and panic disorder in the ED: one of the authors responds to Epstein's letter. PMID- 17466125 TI - Sars. PMID- 17466126 TI - Epinephrine in digital blocks: refuting the refutation of dogma [The author responds:]. PMID- 17466127 TI - Accuracy of clinical diagnosis versus the World Health Organization case definition in the Amoy Garden SARS cohort. AB - OBJECTIVES: To compare the diagnostic accuracy of emergency department (ED) physicians with the World Health Organization (WHO) case definition in a large community-based SARS (severe acute respiratory syndrome) cohort. METHODS: This was a cohort study of all patients from Hong Kong's Amoy Garden complex who presented to an ED SARS screening clinic during a 2-month outbreak. Clinical findings and WHO case definition criteria were recorded, along with ED diagnoses. Final diagnoses were established independently based on relevant diagnostic tests performed after the ED visit. Emergency physician diagnostic accuracy was compared with that of the WHO SARS case definition. Sensitivity, specificity, predictive values and likelihood ratios were calculated using standard formulae. RESULTS: During the study period, 818 patients presented with SARS-like symptoms, including 205 confirmed SARS, 35 undetermined SARS and 578 non-SARS. Sensitivity, specificity and accuracy were 91%, 96% and 94% for ED clinical diagnosis, versus 42%, 86% and 75% for the WHO case definition. Positive likelihood ratios (LR+) were 21.1 for physician judgement and 3.1 for the WHO criteria. Negative likelihood ratios (LR-) were 0.10 for physician judgement and 0.67 for the WHO criteria, indicating that clinician judgement was a much more powerful predictor than the WHO criteria. CONCLUSIONS: Physician clinical judgement was more accurate than the WHO case definition. Reliance on the WHO case definition as a SARS screening tool may lead to an unacceptable rate of misdiagnosis. The SARS case definition must be revised if it is to be used as a screening tool in emergency departments and primary care settings. PMID- 17466128 TI - SARS: finding a deadly needle in the haystack. PMID- 17466129 TI - What is the risk of bacterial meningitis in infants who present to the emergency department with fever and pyuria? AB - OBJECTIVE: To determine the rate of bacterial meningitis among febrile infants in the emergency department (ED) who have pyuria detected in an initial catheterized urine specimen. METHODS: This retrospective chart review, conducted at the Hospital for Sick Children, Toronto, Ont., involved all children aged 0 to 3 months who presented to the ED with fever and pyuria (>/=10 white blood cells/mm(3)) over a 3-year period. Cerebrospinal fluid (CSF) was evaluated using standard methods, and the rate of meningitis in children with pyuria was determined. RESULTS: The study sample included 211 infants with fever and pyuria - 79 of these under 1 month of age. Eighty-one percent (171/211) had positive urine cultures, and 143 underwent lumbar puncture to rule out meningitis. Of these, 140 CSF samples were culture negative and 3 grew coagulase negative Staphylococcus -- 2 because of contamination and 1 because of a ventriculo peritoneal shunt infection. Both children with CSF contamination grew Escherichia coli in the urine. The rate of bacterial meningitis in the study sample was 0% (95% confidence interval, 0%-2.6%). CONCLUSIONS: In this study of febrile children under 90 days of age with fever and pyuria, the incidence of concurrent meningitis was 0%. This suggests that recommendations for mandatory lumbar puncture in such children should be reconsidered. However, until larger prospective studies define a patient subset that does not require CSF analysis, it is prudent to rule out meningitis, administer parenteral antibiotics for urinary tract infection, and admit for close observation. PMID- 17466130 TI - Off-service residents in the emergency department: the need for learner centredness. AB - OBJECTIVES: Standard learning objectives enable residency directors to develop effective programs and evaluate residents based on key goals and parameters. While standards are important for ensuring basic competence, the usual process has little flexibility to address the unique needs and desires of a given resident. Our objective was to determine whether the expectations of off-service residents rotating through an emergency department (ED) rotation were being met. METHODS: We developed a 144-item questionnaire using a 5-point Likert scale and surveyed 25 off-service residents at the beginning and end of their ED rotation. The survey was divided into 3 sections: presentations, skills and diagnoses. RESULTS: The results demonstrate that certain expectations are consistently underachieved while others represent individual variations. CONCLUSION: We propose a learner-centred approach to ensure an optimal emergency educational experience for all trainees. PMID- 17466131 TI - Systematic reviews in emergency medicine: Part II. Critical appraisal of review quality, data synthesis and result interpretation. AB - Reviews of the medical literature have always been an important resource for physicians. Increasingly, qualitative and quantitative systematic reviews (SRs) have replaced the traditional "narrative review" as a means of capturing and summarizing current evidence on a topic or, when possible, answering a specific clinical question. This paper is Part II of a 2-part series designed to provide emergency physicians with the background necessary to locate, critically evaluate and interpret SRs. The paper expands on the critical appraisal principles discussed in Part I by focusing on quality assessment, data synthesis and interpretation of results. To illustrate key points and facilitate readability, examples from the emergency medicine literature have been included and technical details have been kept to a minimum. The references, however, are comprehensive and provide a resource for readers seeking further information. PMID- 17466132 TI - Dexamethasone in acute bacterial meningitis. PMID- 17466133 TI - Benign pneumatosis intestinalis: a cause of massive pneumoperitoneum in the adult. AB - Pneumatosis intestinalis (gas in the bowel wall) is often a benign condition, but it may mimic bowel ischemia or infarction and lead to unnecessary surgical intervention, especially when associated with pneumoperitoneum. We present a case of benign pneumatosis intestinalis with massive pneumoperitoneum and discuss various distinguishing features that may aid in its diagnosis. PMID- 17466134 TI - Acute allergic reaction due to the administration of fibrinolytic therapy for ST segment elevation myocardial infarction: case report and discussion. AB - Although a rare phenomenon, acute allergic reactions to fibrinolytic and heparin therapy have been described in the literature. We report the case of a 63-year old woman who experienced a severe anaphylactic reaction while undergoing fibrinolytic therapy with tissue plasminogen activator for an ST-segment elevation myocardial infarction. Overall outcome was successful, but patient morbidity was increased because of the reaction and the subsequent therapy administered. PMID- 17466135 TI - High times at Kunde Hospital, Nepal. PMID- 17466136 TI - Conflict in the emergency department: Retreat in order to advance. AB - Conflict that exists within a team providing services in an emergency department has the potential to be destructive to clinical care and professional relationships. A collaborative approach involving members of the team guided by a neutral facilitator can be effective in addressing important issues and creating solutions that are acceptable to the individuals and the group. By working together in such a facilitated setting, relationships between the team members can be strengthened while problems are being solved. PMID- 17466137 TI - Chest pain and panic disorder in the ED? PMID- 17466138 TI - Prospective evaluation of clinical assessment in the diagnosis and treatment of clavicle fracture: Are radiographs really necessary? AB - INTRODUCTION: Current recommended treatment for middle-third clavicle fractures is limited to the use of ice, analgesics, a sling, and rest. Radiography for these fractures would be superfluous if physicians could accurately identify them by clinical examination alone. The primary purpose of this study was to determine whether emergency physicians can accurately diagnose clavicle fractures, and whether they can differentiate middle-third fractures from medial- or lateral third fractures by clinical assessment alone. METHODS: We enrolled a convenience sample of patients who presented to our rural emergency department with possible clavicle fracture between Nov. 1, 2001, and Apr. 30, 2002. Prior to viewing radiographs, physicians scored their clinical certainty of diagnosis on a 10-cm visual analogue scale. When certain of fracture, physicians determined the location of the fracture, the nature of the fracture and their hypothetical comfort in treating the injury without radiography. RESULTS: In 51 of 77 enrolled patients (66%; 95% confidence interval [CI], 54.6%-76.6%), treating physicians were certain of the diagnosis of clavicle fracture prior to radiography. In these 51 cases, radiography revealed a fracture in 50 cases (98.0%; 95%CI, 89.6% 99.9%). The physicians were 100% accurate for 4 fractures clinically identified as lateral-third fractures (95% CI, 39.7%-100%) and for 41 fractures identified as middle-third fractures (95% CI, 91.4%-100%). They were correct on only 1 of 5 injuries (20%; 95% CI: 1%-72%) they clinically identified as medial-third fractures. Despite high clinical accuracy with middle-third fractures, they stated in 27 of 42 cases (64%; 95%CI, 48.0%-78.5%) that they would have been uncomfortable treating the patient without a radiograph. CONCLUSIONS: This study provides evidence that experienced emergency physicians are highly accurate when they are clinically certain of clavicle fracture. Further, when emergency physicians do clinically diagnose clavicle fracture, they can accurately identify the patient subgroup that will be responsive to conservative treatment. Routine radiography of obvious middle-third clavicle fractures does not appear to improve diagnostic accuracy or treatment decisions. PMID- 17466139 TI - Implementation of the Canadian Emergency Department Triage and Acuity Scale (CTAS) in the Principality of Andorra: Can triage parameters serve as emergency department quality indicators? AB - OBJECTIVE: To assess the performance of the newly implemented Canadian Emergency Department Triage and Acuity Scale (CTAS) triage system in a redesigned 200-bed community hospital emergency department (ED) and to evaluate the predictive validity of CTAS in this setting. METHODS: Triage system performance was analyzed on the basis of 4 quality indicators: time to triage; triage duration; proportion of patients who left without being seen by a physician; and waiting time to nurse and physician, stratified by triage level and reported as fractile response rates. The predictive validity of CTAS was evaluated by investigating the relationship between CTAS level, hospitalization index, ED length of stay (LOS) and diagnostic test utilization. RESULTS: During the study period, 32 574 patients were triaged and 32 261 were eligible for study. Eighty-five percent were triaged within 10 minutes, and 98% had a triage duration of < 5 minutes. Waiting times to nurse and physician were within CTAS time objectives in 96.3% and 92.3% of cases respectively. The left without being seen (LWBS) rate was 0.96%. Hospitalization rates were compatible with CTAS standards for adults in Levels I, II, III and V and for children in Level V. Median LOS and laboratory test utilization were highly correlated with CTAS Levels II to V (p < 0.01), and similar correlation between triage acuity and imaging utilization was noted in adult patients with non-traumatic non-musculoskeletal complaints (p < 0.01). CONCLUSIONS: The CTAS is adaptable to countries beyond Canada and its operating objectives are achievable. Time to triage and fractile response rates can be considered indicators of triage quality and ED performance. CTAS is a valid instrument for predicting admission rates, hospital LOS and diagnostic utilization. PMID- 17466140 TI - Inter-rater reliability of a computerized presenting-complaint-linked triage system in an urban emergency department. AB - BACKGROUND: Triage reliability studies typically use hypothetical scenarios and weighted kappa scores where agreement within one level is considered satisfactory. But if triage category is used to help define ED case-mix groups for comparative or benchmarking processes, agreement on exact triage level and major system involved is important. Our hypothesis was that a computerized menu that links presenting complaints to preferred triage levels (PC-linked triage) would provide high triage reliability. OBJECTIVES: Our objective was to assess inter-rater reliability of PC-linked triage using the Canadian Emergency Department Triage and Acuity Scale (CTAS) in a real-time clinical setting, considering agreement on exact triage level and primary body system involved. METHODS: On duty triage nurses entered patient presenting complaint and PC-linked triage level as per standard procedure. In a convenience sample of patients, a second nurse, blinded to triage assignment, observed the triage interaction and independently entered presenting complaint and triage level on a dummy terminal. RESULTS: During the study, 15 nurse pairs triaged 266 patients. Study patients matched actual emergency department case mix closely. Triage nurses agreed exactly in 74% of cases and within one level in 94% of cases. The unweighted kappa value was 0.66 (95% confidence interval [CI], 0.60-0.73) and the quadratic weighted kappa value was 0.75 (95% CI, 0.68-0.81). Kappa for agreement on major system involved was 0.80 (95% CI, 0.69-0.91). CONCLUSION: PC-linked triage has high inter-rater reliability in a real-time clinical setting. PC-linked triage may be useful as one factor in defining case-mix groups for benchmarking and comparative purposes. PMID- 17466141 TI - Systematic reviews in emergency medicine: Part I. Background and general principles for locating and critically appraising reviews. AB - Reviews of the medical literature have always been an important resource for physicians. Increasingly, qualitative and quantitative "systematic reviews" have replaced the traditional "narrative review" as a means of capturing and summarizing current evidence on a topic or, when possible, answering a specific clinical question. This paper is part one of a two-part series designed to provide emergency physicians with the background necessary to locate, critically evaluate and interpret systematic reviews. The paper provides a brief background on systematic reviews and general principles on locating and critically appraising them. To facilitate readability, examples from the emergency medicine literature have been included for illustrative purposes and technical details have been kept to a minimum. The references, however, are comprehensive and provide a resource for readers seeking further information. PMID- 17466142 TI - Emergency medicine health advocacy: foundations for training and practice. AB - Emergency physicians (EPs) are uniquely positioned to act as health advocates for individual patients, emergency department (ED) patient populations and the Canadian public. However, most ED practice environments do not encourage health advocacy, and staff EPs often do not feel adequately prepared to address many health-determinant issues. The mandate to provide health advocacy training to emergency medicine residents must be addressed in light of these challenges. This report defines the role of EPs as health advocates and summarizes the advantages and disadvantages of the ED as a forum for advocacy. At the University of Toronto, we have developed a new curriculum using evidence-based ED initiatives, examples of Canadian EP advocacy, and a description of organizations involved in advocacy, and we have incorporated several principles of adult learning to increase learner investment, maximize relevancy for EPs and optimize retention into practice. Residents believe the curriculum is highly relevant, allowing them to recognize advocacy opportunities in their own practices. PMID- 17466143 TI - Implications of the SARS outbreak for Canadian emergency departments. PMID- 17466144 TI - Myth: Cerebrospinal fluid analysis can differentiate bacterial meningitis from aseptic meningitis. PMID- 17466145 TI - Quality of care in walk-in clinics, family practices and emergency departments: the Ontario Walk-In Clinic Study. PMID- 17466146 TI - Hypokalemic thyrotoxic periodic paralysis: a case series. AB - Hypokalemic periodic paralysis is a rare and dramatic complication of hyperthyroidism. This series summarizes the clinical and metabolic features of 10 patients who presented to the Western and Sunshine hospitals in Melbourne, Australia, between 1997 and 2002 with thyrotoxic periodic paralysis (TPP). TPP classically presents with proximal lower-limb weakness in the setting of a low potassium level and biochemical evidence of thyrotoxicosis: low thyroid stimulating hormone levels along with elevated free thyroxine (FT(4)) or free triiodothyronine (FTL(3)). The challenge for emergency physicians is to recognize the association with thyroid disease, since features of hyperthyroidism may not be apparent on history and examination. Acute treatment with potassium supplements and long-term management is aimed at achieving an euthyroid state. Thyrotoxic periodic paralysis is more common in Asian populations; however, increasing immigration from Asia will lead to higher TPP prevalence in Western countries. PMID- 17466147 TI - An attractive approach to magnets adherent across the nasal septum. AB - We report a case of an 8-year-old boy who presented to the emergency department with small jewelry magnets adherent across his nasal septum. Prompt removal of these foreign bodies is important to avoid septal necrosis and perforation. We report our success in using the metal handle of bayonet forceps to break the attraction between the magnets. The magnets were removed painlessly and without trauma to the nasal septum. PMID- 17466148 TI - Emergency medicine on the high seas. PMID- 17466149 TI - Knock, knock, knockin' on heaven's door. PMID- 17466150 TI - Knock, knock, knockin' on heaven's door - Answer. PMID- 17466152 TI - Management of the overactive bladder: a review of pharmacological therapies and methods used by the urological specialist. AB - Overactive bladder is a common urological diagnosis, which is often untreated as patients fail to seek help for this embarrassing problem. This disorder causes significant lifestyle limitations for the patient and is also expensive as it reduces national productivity and therefore affected patients should be treated. It is simple for primary care providers to make a working diagnosis in this disorder and they should be carrying out simple investigations in order to make the diagnosis. Commencement of therapy should start with conservative measures such as lifestyle modifications including pelvic floor exercises and bladder drill followed by the introduction of pharmacological treatments if necessary. The patient should be fully educated about their disorder and about the potential side effects of the medication they are given in order to improve compliance. There are a number of antimuscarinics available on the market for the treatment of overactive bladder but it is often difficult to decide which is the best form of management for these patients. In this review we address the necessary investigations that need to be carried out as well as providing an overview of the different non-surgical and medical treatments for this common problem. Should these therapies fail, then the referral to a urological specialist should be made prior to invasive therapy. PMID- 17466153 TI - Correlation of CT scan versus plain radiography for measuring urinary stone dimensions. AB - OBJECTIVES: To correlate the measured dimensions of urinary stones from spiral non-contrast computerized tomography (CT) with that of plain radiography (KUB). METHODS: The transverse diameter as reported on CT was compared to the measured transverse diameter on KUB for 61 stones. The transverse and craniocaudal dimensions on CT were then re-measured for 30 urinary stones and again compared to the re-measured values for KUB. The craniocaudal dimension on CT was determined by measuring the stone on reconstructed coronal CT images. Measurements between imaging modalities were blinded and performed consecutively by a dedicated investigator. RESULTS: The mean transverse size of the stones on the initial CT report was 6.0 mm +/- 2.8 mm versus 5.6 mm +/- 2.3 mm on KUB (paired t-test, p = 0.05, 95% CI difference between the means -1.3 to 0.5). The stones were categorized in transverse size ranges of 1.0 mm to 5.0 mm, > 5.0 mm to 10.0 mm, and > 10.0 mm. A total of 14 stones failed to be put into the same size categories by the two methods. The largest difference in measurements was 5 mm. In the second analysis, where the CT dimensions were re-measured, the mean transverse dimension on CT was 4.5 mm +/- 2.1 mm versus 4.7 mm +/- 2.0 mm on plain radiography (paired t-test, p = 0.06, 95% CI difference between the means 0.02 to 0.6). Mean craniocaudal dimension of the stones on CT was 7.4 mm +/- 3.2 mm versus 6.0 mm +/- 2.7 mm on plain radiography (paired t-test, p = 0.0001, 95% CI between the means -2.0 to -0.9). When the stones were categorized in transverse size ranges of 1.0 mm to 5.0 mm, >5.0mm to 10.0mm, and >10.0mm, CT and KUB agreed for 30/30 stones. CONCLUSIONS: In this study, the initially reported CT transverse values were found to be significantly different from measured KUB values; moreover, large differences of up to 5 mm were found between the measurements. With fastidious measurement of stone dimensions on both CT and KUB, we found that the transverse dimension of stones measured by the two imaging modalities were similar. The craniocaudal measurements of the stones were found to be significantly different on CT versus KUB, with CT measurement being 1.4 mm larger on average. PMID- 17466154 TI - Percutaneous bladder catheterization in microgravity. AB - INTRODUCTION: Urinary obstruction (UO) or failure to void has been observed during several episodes of short-duration spaceflight, necessitating bladder catheterization. It should be considered a possible medical condition in long duration space missions as well. Antiemetics used early in space flight add to the risk and severity of voiding problems, along with the sensory and psychological peculiarities of voiding without gravity and in the unusual setting of a spacecraft. Urolithiasis due to the above-normal calcium excretion increases the risk of UO in long duration space missions. Finally, the individual risk of UO is higher against the background of preexisting conditions such as benign prostatic hyperplasia (BPH) or urethral stricture. Both acute retention and ureteral obstruction are associated with substantial patient distress, and carry a risk of urosepsis and/or acute renal failure. If UO in orbital flight is unresolved or complicated, it would likely result in crew emergency return from orbit. Exploration missions, however, may require means for definitive treatment of urinary tract obstruction. This study documents successful ultrasound-guided percutaneous catheterization of the urinary bladder in microgravity. A porcine model of urethral occlusion was used. The results demonstrate an additional capability from our previous investigations describing endoscopic catheterization and stenting of the ureters in microgravity conditions. METHODS: In an anesthetized porcine model, a Foley catheter was placed in the bladder and clamped after instillation of 200 ml of colored liquid. The bladder was visualized and then drained under ultrasound guidance through suprapubic puncture, employing a 10.3 F pigtail catheter with introducer. The procedural elements were conducted only during microgravity portions of the parabolic flight. RESULTS: Ultrasound imaging was used to successfully perform image-guided percutaneous puncture through the anterior bladder wall with the catheter, without injury to adjacent organs. The percutaneous catheter was able to successfully drain the bladder in microgravity conditions. CONCLUSIONS: Percutaneous bladder catheterization and drainage can be successfully performed in weightless conditions under ultrasound guidance. Ultrasound provides a low power, portable means to safely conduct minimally invasive procedures in pertinent organs and tissues. Percutaneous bladder catheterization is a standard procedure when luminal bladder catheterization is not possible; this technique can be successfully modified for use in space medicine applications. PMID- 17466155 TI - Mechanical failure rate of da Vinci robotic system. AB - INTRODUCTION: Robotic-assisted laparoscopic radical prostatectomy (RLRP) is playing an increasing role in the surgical management of prostate cancer. The benefits of minimally invasive surgery, enhanced surgeon familiarity with the instrumentation, and increased patient demand has led to the popularity of this surgical technique. There are, however, shortcomings specifically associated with this technology. Notably, instrumentation failure associated with robotic procedures represents a new and unique problem in urological surgery. We examine the rate of mechanical failure of the da Vinci robotic system and its impact on our prostate cancer program. MATERIALS AND METHODS: We reviewed our prospective, institutional review board-approved database of the first 350 RLRP procedures that were scheduled for surgery at our institution. We identified all cases in which mechanical failure of the da Vinci robotic system resulted in surgery being cancelled, postponed, or converted to a conventional laparoscopic or an open radical prostatectomy. RESULTS: Nine of the 350 (2.6%) scheduled RLRPs were unable to be completed robotically secondary to device malfunction. Six of the malfunctions were detected prior to anesthesia induction and surgery was rescheduled. Three other malfunctions occurred intraoperatively and were converted either to a conventional laparoscopic (1 case) or an open surgical approach (2 cases). The etiology of the malfunctions included the following: set up joint malfunction (2), arm malfunction (2), power error (1), monocular monitor loss (1), camera malfunction (1), metal fatigue/ break of surgeon's console hand piece (1) and software incompatibility (1). CONCLUSIONS: Although uncommon, malfunction of the da Vinci robotic system does occur and may lead to psychological, financial, and logistical burdens for patients, physicians, and hospitals. Patients should be carefully counseled preoperatively regarding the possibility of robotic mechanical failure. PMID- 17466156 TI - Adenocarcinomas of the prostatic duct in necropsy material. AB - The general consensus about prostatic duct adenocarcinomas is that they have a rather aggressive biological behavior. In addition, studies or reports of latent adenocarcinoma of the prostatic duct in necropsy material are scarce in the literature. We report here three cases of adenocarcinoma of the prostatic duct that were found incidentally among 39 cases of latent acinar prostate adenocarcinomas in necropsy material. We examined the morphologic and histological features of these prostatic duct adenocarcinomas, in order to better understand their biological behavior. We identified two cases of mixed ductal acinar adenocarcinoma and one case of pure ductal adenocarcinoma. The pure form had a favorable histological differentiation, while the mixed forms had intermediate histological differentiation patterns. Invasiveness was related to both volume and histological differentiation. The finding of prostatic ductal adenocarcinomas among autopsy material, as well as some of their histological features, suggest that these tumors might have a similar biological potential as prostatic acinar cancer. PMID- 17466157 TI - Diffuse xanthogranulomatous pyelonephritis with psoas abscess in a pregnant woman. AB - We report the first case, to our knowledge, of xanthogranulomatous pyelonephritis (XGP) with psoas abscess occurring during pregnancy. A 37-year-old woman in the third trimester of pregnancy presented with low back pain and a fever. From sonographic features, a multidisciplinary team decided to perform computed tomography of the abdomen with contrast agent injection, which strongly suggested diffuse XGP of the left kidney with a psoas abscess. Cesarean section at 32 weeks was followed by extended nephrectomy. Pathological examination of the operative specimen confirmed the diagnosis. Outcomes were favorable in the mother and baby. The diagnosis and treatment of XGP during pregnancy are discussed. PMID- 17466158 TI - Malignant transformation in abdominal cryptorchid testis 40 years after treatment of seminoma confined to other testis. AB - This report describes an exceptionally rare case of a 64-year-old man with seminoma in abdominal cryptorchidism, leading to intestinal obstruction 40 years after curative treatment for seminoma of the other (descended) testis. PMID- 17466159 TI - Malignant mesothelioma of the tunica vaginalis. AB - INTRODUCTION: Malignant mesothelioma involving the tunica vaginalis is an extremely rare tumor. CASE: We describe a case of a 39-year-old man who initially presented with discomfort in the left testis and underwent resection of a hydrocele in the left testis. A pathology examination of a frozen section of a lesion on the tunica identified the lesion as mesothelioma, which was confirmed by a second pathology examination. No metastatic disease was found. The patient subsequently underwent a left radical orchiectomy and a partial scrotectomy, and has been disease free for 7 years. DISCUSSION: The first case of malignant mesothelioma involving the tunica vaginalis, a structure embryologically derived from a layer of reflected peritoneum known as the "processus vaginalis," was reported in 1957. Only 73 cases were reported in the literature between 1966 and 1997. In up to 41% of cases, there is a positive history of asbestos exposure. Young age and localized disease are associated with a better prognosis. Radical orchiectomy limits recurrence, which usually occurs within 2 years, but may occur up to 15 years after surgery. PMID- 17466160 TI - 1500 gram suprarenal mass: a case report. AB - Adrenocortical carcinoma can have a clinical presentation that mimics a primary renal tumor. We describe a case of a 47-year-old male who presented with flank pain, weight loss, and a 14 cm mass arising from the upper pole of the right kidney on imaging. Upon surgical resection he was found to have a 1500 gram stage II adrenocortical carcinoma. The clinical features, pathologic findings, grading criteria, and differential diagnosis of adrenocortical carcinoma are reviewed. PMID- 17466161 TI - BCG pyelonephritis following intravesical therapy for transitional cell carcinoma. AB - Bacillus Calmette-Guerin (BCG) is a common intravesical agent utilized for superficial transitional cell carcinoma of the bladder. The potential for side effects and complications does exist secondary to the utilization of live attenuated bacteria. We present the case of a 70-year old male that developed BCG pyelonephritis and associated renal mass following the fifth of six weekly intravesical BCG therapies. The patient improved clinically and renal mass resolved following 3 months of isoniazid monotherapy without renal biopsy. PMID- 17466162 TI - Ureteric obstruction: an unusual presentation of metastatic colon carcinoma. AB - We present the case of a 78-year-old male who presented to clinic for follow-up of a papillary transitional cell carcinoma of the urinary bladder. Notably, the patient also had a history of colorectal resection for an adenocarcinoma. The follow-up appointment revealed left hydronephrosis with evidence of a distal ureteric stricture. Cytology and biopsy from the ureter subsequently disclosed the presence of malignant cells that were originally thought to be of urothelial origin. Upon surgical resection the lesion was found to be an adenocarcinoma, morphologically consistent with a metastasis from the patient's primary colonic adenocarcinoma. This case illustrates a diagnostically challenging situation, with metastatic colonic carcinoma to the ureter occurring in a patient with two previously documented malignancies. PMID- 17466195 TI - Standardized colonoscopy reporting and data system: report of the Quality Assurance Task Group of the National Colorectal Cancer Roundtable. AB - BACKGROUND: Standardized reporting systems for diagnostic and screening tests facilitate quality improvement programs and clear communication among health care providers. Although colonoscopy is commonly used for screening, diagnosis, and therapy, no standardized reporting system for this procedure currently exists. The Quality Assurance Task Group of the National Colorectal Cancer Roundtable developed a reporting and data system for colonoscopy based on continuous quality improvement indicators. DESIGN: The Task Group systematically reviewed quality indicators recommended by the Multi-Society Task Force on Colorectal Cancer and developed consensus-based terminology for reporting and data systems to capture these data elements. The Task Group included experts in several disciplines: gastroenterology, primary care, diagnostic imaging, and health care delivery. RESULTS AND CONCLUSIONS: The standardized colonoscopy reporting and data system provides a tool that can be used for efforts in continuous quality improvement within and across practices that use colonoscopy. PMID- 17466196 TI - Appropriateness of upper-GI endoscopy: an Italian survey on behalf of the Italian Society of Digestive Endoscopy. AB - BACKGROUND AND OBJECTIVE: Open-access endoscopy allows physicians to directly schedule endoscopic procedures for their patients without prior consultation. An evaluation of both appropriateness and diagnostic yield of endoscopic procedures is critical when assessing the costs and benefits of endoscopy in an open-access setting. The aim of this Italian multicenter study was to assess the appropriate use of upper endoscopy (EGD) in an open-access system and to establish the yield of diagnostic information relevant to patient care. DESIGN AND SETTING: Cross sectional, prospective, multicenter study. PATIENTS: A total of 6270 patients referred to 44 Italian centers for open-access EGD during 1 month were prospectively enrolled. INTERVENTIONS: The American Society for Gastrointestinal Endoscopy (ASGE) guidelines were used to assess the relation between the appropriate use of EGD and the presence of relevant endoscopic findings. MAIN OUTCOME MEASUREMENTS AND RESULTS: The rate for "generally not indicated" EGDs was 22.9%: 29.4% for primary care physicians and 12.9% for specialists (P < .01). A relevant endoscopic finding was detected in 2929 examinations (46.7%). The diagnostic yield was significantly higher for "generally indicated" EGDs compared with "generally not indicated" procedures (52% vs 29%; odds ratio [OR] 2.65, 99% confidence interval [CI] 2.23-3.20; P < .01). Of the 133 malignant lesions diagnosed, all but 1 were diagnosed in patients with an appropriate indication (OR >20, 99% CI 3 to >100; P < .01). CONCLUSIONS: Open-access EGD is an useful procedure for clinical practice. Because most of the relevant findings were detected during examinations performed for appropriate indications, the use of ASGE guidelines emerges as crucial to the cost-effectiveness of an open-access system. PMID- 17466197 TI - Patient satisfaction scores for endoscopic procedures: impact of a survey collection method. AB - BACKGROUND: Many endoscopy units administer patient satisfaction surveys. We hypothesized that the survey collection method would affect satisfaction scores. OBJECTIVE: To compare satisfaction scores obtained by using on-site (OS) surveys versus mail-back (MB) surveys. DESIGN: Quasi-randomization based on alternating weeks. SETTING: Teaching hospital. PATIENTS: Patients undergoing elective routine outpatient colonoscopy or upper endoscopy. INTERVENTIONS: Every patient was given an 11-question survey that asked about the patient's satisfaction with the nurses and the physician, wait times, the bowel-preparation process, patient education, procedural comfort, and sedation. Survey collection methods alternated weekly between an OS versus an MB method. MAIN OUTCOME MEASUREMENTS: Satisfaction scores on a Likert scale ranged from 1 (worst) to 7 (best). RESULTS: A total of 1698 subjects were included. The response rate was higher for the OS group (95%) than the MB group (62%). OS scores were significantly higher than MB scores for 5 of 11 questions, which concerned nurse satisfaction, physician satisfaction, bowel preparation comfort, postprocedure education, and overall satisfaction (Bonferroni adjusted P < .05 for all). Younger patients gave lower scores than older patients for all questions, whereas women gave significantly lower scores than men for bowel-preparation satisfaction. LIMITATIONS: Lack of true randomization and formal validation of the satisfaction survey. CONCLUSIONS: Survey collection methods may bias not only response rates but also satisfaction scores. OS survey collection methods tend to result in higher satisfaction scores than MB methods. This bias should be noted when comparing scores among studies that used different survey collection methods. PMID- 17466198 TI - EUS criteria for vascular invasion: analyzing the meta-analysis. PMID- 17466199 TI - Diagnostic performance of EUS for chronic pancreatitis: a comparison with histopathology. AB - BACKGROUND: EUS has been proposed as a minimally invasive and accurate test to detect chronic pancreatitis (CP). OBJECTIVE: To investigate the correlation between EUS criteria and histopathology grading in patients with presumed CP. DESIGN: Retrospective study. SETTING AND PATIENTS: Patients who received pancreatic surgery according to presumed CP from the Medical University of South Carolina surgical database between 1995 and 2003 were identified and included if EUS was performed within 1 year before surgery. The number of EUS criteria for CP was compared with a histologic fibrosis score (FS). MAIN OUTCOME MEASUREMENTS: Sensitivity and specificity of number of EUS criteria compared with FS. RESULTS: Seventy-one patients were identified (38 women). Median FS was 7 (range, 0-12). Of the patients with calcifications: calcifications were detected by EUS in 30 (42%), 14 (47%) had calcifications missed by other imaging modalities, and 28 (93%) were confirmed to have abnormal histology (FS > or = 2). Of the patients without calcifications: in the 41 patients without calcifications on EUS, 36 (88%) had FS > or = 2; median FS was 5 (range, 0-12); the correlation between the number of EUS criteria and FS was low but statistically significant (r = 0.40; P = .01). Three or more EUS criteria provided the best balance of sensitivity (83.3%) and specificity (80.0%) for predicting abnormal histology. LIMITATIONS: Retrospective study. All patients were believed to need surgery. CONCLUSIONS: A threshold of 3 or more EUS criteria provides the best balance of sensitivity and specificity for histologic pancreatic fibrosis. Calcifications seen by EUS but missed by other imaging are common in this group of patients. PMID- 17466200 TI - Diagnosing chronic pancreatitis: the enemy of "good" is "better". PMID- 17466201 TI - Antireflux biliary stents: is it time to go with the flow? PMID- 17466202 TI - SpyGlass single-operator peroral cholangiopancreatoscopy system for the diagnosis and therapy of bile-duct disorders: a clinical feasibility study (with video). AB - BACKGROUND: Clinical implementation of cholangioscopy for direct visual examination of bile ducts, tissue sampling, and therapeutic maneuvers has been slowed by limitations in available technology. With 4-way deflected steering and dedicated irrigation channels, the single-operator SpyGlass peroral cholangiopancreatoscopy system is designed to overcome some of these limitations. OBJECTIVE: To evaluate the clinical utility and safety of the SpyGlass system for diagnostic and therapeutic endoscopic procedures in bile ducts. DESIGN: Prospective observational clinical feasibility study. SETTING: Two tertiary referral centers. MAIN OUTCOME MEASUREMENTS: Procedural success rate defined as the proportion of SpyGlass procedures in which the diagnostic or therapeutic objectives of the procedure were achieved. RESULTS: SpyGlass procedures were performed in 35 patients: 22 with indeterminate strictures (63%), 5 with indeterminate filling defects (14%), 5 with stones (14%), 2 with cystic lesions (6%), and 1 patient with an indication for gallbladder stent placement (3%). The rate of procedural success was 91% (95% confidence interval 77%-98%). Twenty patients underwent SpyGlass-directed biopsy, and the specimens procured from 19 patients (95%) were found adequate for histologic evaluation. The preliminary sensitivity and specificity of SpyGlass-directed biopsy to diagnose malignancy were 71% and 100%, respectively. SpyGlass-directed electrohydraulic lithotripsy succeeded in 5 of 5 patients (100%). Procedure-related complications occurred in 2 patients (6%) and resolved uneventfully. LIMITATIONS: No control group was included. Follow-up for determining preliminary sensitivity and specificity was limited. CONCLUSIONS: SpyGlass procedures proved to be clinically feasible, provided adequate samples for histologic diagnosis, and successfully guided stone therapy. The procedures were safe and well tolerated. Prospective multicenter clinical trials of the system are underway. PMID- 17466203 TI - Endoscopic placement of the small-bowel video capsule by using a capsule endoscope delivery device. AB - BACKGROUND: Capsule endoscopy performed via the traditional peroral route is technically challenging in patients with dysphagia, gastroparesis, and/or abnormal upper-GI (UGI) anatomy. OBJECTIVE: To describe the indications and outcomes of cases in which the AdvanCE capsule endoscope delivery device, which has recently been cleared by the Food and Drug Administration, was used. DESIGN: Retrospective, descriptive, case series. SETTING: Tertiary care, university hospital. PATIENTS: We report a case series of 16 consecutive patients in whom the AdvanCE delivery device was used. The study period was May 2005 through July 2006. INTERVENTIONS: Endoscopic delivery of the video capsule to the proximal small bowel by using the AdvanCE delivery device. MAIN OUTCOME MEASUREMENTS: Indications, technique, and completeness of small bowel imaging in patients who underwent endoscopic video capsule delivery. RESULTS: The AdvanCE delivery device was used in 16 patients ranging in age from 3 to 74 years. The primary indications for endoscopic delivery included inability to swallow the capsule (10), altered UGI anatomy (4), and gastroparesis (2). Of the 4 patients with altered UGI anatomy, 3 had dual intestinal loop anatomy (ie, Bilroth-II procedure, Whipple surgery, Roux-en-Y gastric bypass) and 1 had a failed Nissen fundoplication. In all cases, the capsule was easily deployed without complication, and complete small intestinal imaging was achieved. LIMITATIONS: Small patient size. CONCLUSIONS: Endoscopic placement of the Given PillCam by use of the AdvanCE delivery device was safe and easily performed in patients for whom capsule endoscopy would otherwise have been contraindicated or technically challenging. PMID- 17466204 TI - When cecal intubation is a "must". PMID- 17466205 TI - Short- and long-term outcomes of standardized EMR of nonpolypoid (flat and depressed) colorectal lesions > or = 1 cm (with video). AB - BACKGROUND: Nonpolypoid (flat and depressed) colorectal lesions are increasingly recognized. Their endoscopic removal requires specialized EMR techniques, which are more complex to perform. Outcomes data on EMR of nonpolypoid neoplasms in the United States is needed. OBJECTIVE: To determine the safety and efficacy of EMR in the resection of nonpolypoid colorectal neoplasms > or = 1 cm. DESIGN: Retrospective analysis. SETTING: Veterans Affairs Palo Alto Health Care System. PATIENTS: Over a 5-year period, patients who underwent EMR for nonpolypoid colorectal lesions > or = 1 cm. INTERVENTION: A standardized approach that included lesion assessment, classification, inject-and-cut EMR technique, reassessment, and treatment of residual tissue. MAIN OUTCOME MEASUREMENTS: Complete resection, bleeding, perforation, development of advanced cancer, and death. RESULTS: A total of 100 patients (125 lesions: 117 flat and 8 depressed) met inclusion criteria. Mean size was 16.7 +/- 7 mm (range, 10-50 mm). Histology included 5 submucosal invasive cancers, 5 carcinomas in situ, and 91 adenomas. Thirty-eight patients (48 lesions) did not receive surveillance colonoscopy: 8 had surgery, 16 had hyperplastic pathology, and 14 did not undergo repeat examination. Surveillance colonoscopy was performed on 62 patients (77 lesions). Complete resection was achieved in 100% of these patients after 1 to 3 surveillance colonoscopies. All patients received follow-up (mean [standard deviation] = 4.5 +/- 1.4 years); none developed colorectal cancer or metastasis. LIMITATIONS: Single endoscopist, retrospective study. CONCLUSIONS: A standardized EMR (inject-and-cut) technique is a safe and curative treatment option in nonpolypoid colorectal neoplasms (> or = 1 cm) in the United States. PMID- 17466206 TI - EUS with CT improves efficiency and structure identification over conventional EUS. AB - BACKGROUND: EUS is complicated because of the subtleties of US interpretation, small fields of observation, and uncertainty of probe position and orientation. OBJECTIVE: Improved EUS performance is sought by providing contextual information to support US probe positioning and identification of features in US images. Our aims were to demonstrate the feasibility of the image registered gastroscopic US (IRGUS) system in a porcine model and to compare the effectiveness and the efficiency of IRGUS with traditional EUS. DESIGN: Animal feasibility study. INTERVENTIONS: The IRGUS system uses preprocedure CT and miniature US probe trackers to create real-time synthetic displays of the position of the probe tip and a matched slice of CT data for comparison with the US image. Participants used EUS and IRGUS systems in a porcine model to evaluate the speed and accuracy of structure identification. MAIN OUTCOME MEASUREMENTS: The performance and utility of IRGUS were determined by the number of correctly identified structures in a timed trial, kinematic variables, and a structured survey. RESULTS: IRGUS was twice as effective as EUS in localizing and identifying individual structures. In timed trials, IRGUS users identified over 25% more structures than EUS users. Improvement in examination efficiency and accuracy of feature identification was statistically significant, and 90% of the users preferred IRGUS to EUS for these tasks. CONCLUSIONS: IRGUS appears feasible and may be superior to conventional EUS in efficiency and accuracy of probe positioning and in image interpretation. IRGUS has the potential to shorten the EUS learning curve and to broaden the adoption of EUS techniques by gastroenterologists. PMID- 17466208 TI - Animal laboratory endoscopic research: a fellow's perspective. PMID- 17466207 TI - A journey toward excellence: training future gastroenterologists - The Gastroenterology Core Curriculum, third edition. PMID- 17466209 TI - Under the hood: introducing a new series for Gastrointestinal Endoscopy. PMID- 17466210 TI - The endoscope. PMID- 17466211 TI - Iatrogenic colonic perforations: a threat turned into insignificance? PMID- 17466212 TI - Hepatocellular carcinoma invading the fourth branch of the left intrahepatic duct. PMID- 17466213 TI - Insertion of expandable metallic stents in esophageal cancer without fluoroscopy is safe and effective: a 5-year experience. AB - BACKGROUND: Self-expanding metallic stent (SEMS) placement is an important method of dysphagia palliation for patients with inoperable esophageal cancer. In most institutions, it is performed with fluoroscopic guidance; however, in 2001, we described a novel, direct-vision approach to SEMS placement, which does not require fluoroscopy. Here we report an audit of our experience over the last 5 years when using this methodology. OBJECTIVE: To describe our 5-year experience of 98 patients in whom esophageal stents were inserted when using the direct vision technique and compare outcomes with published series of radiography-guided stents. DESIGN: Retrospective review of single-center experience. SETTING: English National Health Service Cancer Centre Hospital. PATIENTS: All patients who underwent esophageal stent insertion for a primary esophageal malignancy. INTERVENTIONS: SEMS insertion by direct endoscopic vision. RESULTS: Ninety-eight patients underwent SEMS insertion for malignant dysphagia during the study period, 92% of which were inserted without fluoroscopy. The technique had a low complication rate, which was consistent with published fluoroscopic data, and the median survival beyond stent insertion was 100 days (interquartile range, 62, 256; range, 4-921 days). In 59 patients, no further endoscopic palliative procedure was required. LIMITATIONS: Retrospective data collection. CONCLUSIONS: This series confirms direct-vision SEMS placement as a safe and efficacious method of malignant dysphagia palliation, providing definitive treatment in almost two thirds of cases. PMID- 17466214 TI - Insertion of expandable metallic stents in esophageal cancer without fluoroscopy: is it safe? PMID- 17466215 TI - Pancreatogastric fistulas associated with chronic pancreatitis: a case report and review of the literature. PMID- 17466216 TI - Metastatic prostate cancer presenting as mediastinal lymphadenopathy identified by EUS with FNA. PMID- 17466217 TI - Is endoscopic screening warranted before major surgical procedures? PMID- 17466218 TI - Maximizing control of tip deflection with sound ergonomics: the "left hand shaft grip". PMID- 17466219 TI - Endoscopic treatment of esophageal food impactions. PMID- 17466222 TI - [Heterotopic pregnancy: a case report with a rare symptomatology]. AB - Heterotopic pregnancy was a rare pathology many years ago, but its survey is more frequently observed during last years with Assisted Reproduction Technics. We report a case of a heterotopic pregnancy with a rare symptomatology, with in the front plan a miscarriage. It's diagnostic is difficult and occurred very late. The main treatment consists to perform a salpingotomy under laparoscopy. However laparotomy treatment is frequently performed in relation to the late time of diagnosis. PMID- 17466223 TI - [Pregnancy in a patient with Gitelman syndrome: a case report and review of literature]. AB - Gitelman syndrome (GS) is a tubulopathy characterized by hypokaliemia, hypomagnesiemia, metabolic alkalosis and hypocalciuria. We report a case of a 33 year-old pregnant woman with Gitelman Syndrome. Oral potassium chloride and magnesium citrate were prescribed and the course of the pregnancy was uneventful with vaginal delivery at term. The impact of GS on the physiologic adaptations to pregnancy is not well-known, with few reports to date. Monitoring of serum potassium and magnesium levels with supplementation, amniotic fluid and fetal growth is required to prevent obstetrical and fetal complications in a patient with GS. PMID- 17466224 TI - Safety of aggressive lipid management. AB - Data from recent clinical trials of high- versus moderate-dose statin therapy support the recommendation to achieve a low-density lipoprotein (LDL) <100 mg/dl in high-risk patients and reveal that many patients will require a high-dose statin to achieve this goal. Overall, low rates of serious musculoskeletal (<0.6%) and hepatic (<1.3%) toxicity have been observed with high-dose statin therapy. In the long-term trials, atorvastatin 80 mg had higher rates of persistent transaminase elevations but rates of myopathy and rhabdomyolysis similar to lower doses of statins. The rate of myopathy and rhabdomyolysis for simvastatin 80 mg, although still low, was about 4x higher than for atorvastatin 80 mg and lower doses of statin. A similar margin of safety would be expected in properly selected patients with characteristics similar to those who participated in the clinical trials. High-dose statin therapy or combination therapy will be required for the large majority of very high-risk patients to achieve the optional LDL goal of <70 mg/dl. While the combination of ezetimibe, bile-acid sequestering agents, niacin, and fenofibrate with moderate dose statins appears to be reasonably safe, the long-term safety of combination with high-dose statins remains to be established. In order to optimize patient outcomes, clinicians should be aware of specific patient characteristics, such as advancing age, gender, body mass index, or glomerular filtration rate, which predict muscle and hepatic statin toxicity. PMID- 17466225 TI - A clinical randomized trial to evaluate the safety of a noninvasive approach in high-risk patients undergoing major vascular surgery: the DECREASE-V Pilot Study. AB - OBJECTIVES: The purpose of this research was to perform a feasibility study of prophylactic coronary revascularization in patients with preoperative extensive stress-induced ischemia. BACKGROUND: Prophylactic coronary revascularization in vascular surgery patients with coronary artery disease does not improve postoperative outcome. If a beneficial effect is to be expected, then at least those with extensive coronary artery disease should benefit from this strategy. METHODS: One thousand eight hundred eighty patients were screened, and those with > or =3 risk factors underwent cardiac testing using dobutamine echocardiography (17-segment model) or stress nuclear imaging (6-wall model). Those with extensive stress-induced ischemia (> or =5 segments or > or =3 walls) were randomly assigned for additional revascularization. All received beta-blockers aiming at a heart rate of 60 to 65 beats/min, and antiplatelet therapy was continued during surgery. The end points were the composite of all-cause death or myocardial infarction at 30 days and during 1-year follow-up. RESULTS: Of 430 high-risk patients, 101 (23%) showed extensive ischemia and were randomly assigned to revascularization (n = 49) or no revascularization. Coronary angiography showed 2 vessel disease in 12 (24%), 3-vessel disease in 33 (67%), and left main in 4 (8%). Two patients died after revascularization, but before operation, because of a ruptured aneurysm. Revascularization did not improve 30-day outcome; the incidence of the composite end point was 43% versus 33% (odds ratio 1.4, 95% confidence interval 0.7 to 2.8; p = 0.30). Also, no benefit during 1-year follow up was observed after coronary revascularization (49% vs. 44%, odds ratio 1.2, 95% confidence interval 0.7 to 2.3; p = 0.48). CONCLUSIONS: In this randomized pilot study, designed to obtain efficacy and safety estimates, preoperative coronary revascularization in high-risk patients was not associated with an improved outcome. PMID- 17466226 TI - Coronary revascularization before noncardiac vascular surgery: one more step forward in understanding its role. PMID- 17466228 TI - PROactive in patients with type 2 diabetes and previous myocardial infarction: swinging the sword of Damocles? PMID- 17466229 TI - Coronary sinus reducer stent for the treatment of chronic refractory angina pectoris: a prospective, open-label, multicenter, safety feasibility first-in-man study. AB - OBJECTIVES: This study sought to evaluate the safety of the Coronary Sinus Reducer (Neovasc Medical, Inc., Or Yehuda, Israel) as a potential alternate therapy for patients with refractory angina who are not candidates for conventional revascularization procedures. BACKGROUND: Increased coronary sinus (CS) pressure can reduce myocardial ischemia by redistribution of blood from nonischemic to ischemic territories. The Coronary Sinus Reducer is a percutaneous implantable device designed to establish CS narrowing and to elevate CS pressure. In preclinical experiments, implantation of the Reducer was safe and was associated with improved ischemic parameters. In the present study, the safety and feasibility of the Coronary Sinus Reducer was evaluated in patients with refractory angina who were not candidates for revascularization. METHODS: Fifteen coronary artery disease patients with severe angina and reversible ischemia were electively treated with the Reducer. Clinical evaluation, dobutamine echocardiography, thallium single-photon emission computed tomography, and administration of an angina questionnaire were performed before and 6 months after implantation. Cardiac computed tomography was performed 2 days and 6 months after implantation. RESULTS: All procedures were completed successfully. No procedure-related adverse events occurred during the periprocedural and the follow-up periods. Angina score improved in 12 of 14 patients. Average Canadian Cardiovascular Society score was 3.07 at baseline and 1.64 at follow-up (n = 14, p < 0.0001). Stress-induced ST-segment depression was reduced in 6 of 9 patients and was eliminated in 2 of these 6 (p = 0.047). The extent and severity of myocardial ischemia by dobutamine echocardiography and by thallium single-photon emission computed tomography was reduced (p = 0.004 [n = 13] and p = 0.042 [n = 10], respectively). CONCLUSIONS: Implantation of the Coronary Sinus Reducer is feasible and safe. These findings, along with the clinical improvement observed, support further evaluation of the Reducer as an alternative treatment for patients with chronic refractory angina who are not candidates for coronary revascularization. PMID- 17466230 TI - Characteristics, management, and outcomes of 5,557 patients age > or =90 years with acute coronary syndromes: results from the CRUSADE Initiative. AB - OBJECTIVES: The goal of this work was to explore the treatment and outcomes of patients with non-ST-segment elevation acute coronary syndromes (NSTE-ACS) age > or =90 years. BACKGROUND: The elderly are often excluded from clinical trials of NSTE-ACS and are underrepresented in clinical registries. METHODS: We used data from the CRUSADE registry to study 5,557 patients with NSTE-ACS age > or =90 years and compared their baseline characteristics, treatment patterns, and in hospital outcomes with a cohort age 75 to 89 years (n = 46,270). RESULTS: Although both groups had much in common, compared with the younger elderly, the older elderly were less likely to be diabetic, smokers, or obese. Among patients without contraindications, the older elderly were less likely to receive glycoprotein IIb/IIIa inhibitors and statins during the first 24 h and were less likely to undergo cardiac catheterization within 48 h. The older elderly were more likely to die (12.0% vs. 7.8%) and experienced more frequent adverse events (26.8% vs. 21.3%) during the hospitalization-differences that persisted after adjustment for baseline patient and hospital characteristics. Increasing adherence to guideline-recommended therapies was associated with both increased bleeding and a graded reduction in risk-adjusted in-hospital mortality across both age groups. CONCLUSIONS: In this large population of nonagenarians and centenarians with NSTE-ACS, increasing adherence to guideline-recommended therapies was associated with decreased mortality. These findings reinforce the importance of optimizing care patterns for even the oldest patients with NSTE ACS, while examining novel approaches to reduce the risk of bleeding in this rapidly expanding patient population. PMID- 17466231 TI - Distributions of C-reactive protein and its association with metabolic syndrome in middle-aged and older Chinese people. AB - OBJECTIVES: We evaluated the distributions of C-reactive protein (CRP) and its association with metabolic syndrome (MetS) in middle-aged and older Chinese people. BACKGROUND: Several studies have suggested that CRP is a risk factor of MetS. However, it remains unclear how CRP levels are distributed and whether they are associated with MetS in Chinese people. METHODS: We conducted a population based cross-sectional survey in 2005 in Beijing and Shanghai, with a total of 1,458 men and 1,831 women age 50 to 70 years. Metabolic syndrome was defined according to the updated National Cholesterol Education Program Adult Treatment Panel III criteria for Asian Americans. RESULTS: The median CRP level was 0.68 mg/l among the study population. The CRP levels were significantly higher among participants from Beijing or from urban areas than those in participants from Shanghai or from rural areas (p < 0.01). No gender difference in CRP levels was observed. The prevalence of MetS progressively increased with elevated CRP levels (p < 0.0001 for trend). In the highest quartile of CRP levels (>1.50 mg/l), the risk for MetS was substantially higher (odds ratio 5.97; 95% confidence interval 4.75 to 7.51) compared with that in the lowest quartile of CRP levels (< or =0.33 mg/l) after adjustment for age, gender, geographic location, lifestyle factors, educational attainment, and family history of chronic diseases. This association was observed in both obese and nonobese participants. CONCLUSIONS: The overall plasma level of CRP is low but highly associated with the MetS among the middle aged and elderly Chinese population. Prospective studies are needed to investigate the role of CRP in the development of MetS and related chronic diseases among Chinese people. PMID- 17466227 TI - The effect of pioglitazone on recurrent myocardial infarction in 2,445 patients with type 2 diabetes and previous myocardial infarction: results from the PROactive (PROactive 05) Study. AB - OBJECTIVES: This analysis from the PROactive (PROspective pioglitAzone Clinical Trial In macroVascular Events) study assesses the effects of pioglitazone on mortality and macrovascular morbidity in patients with type 2 diabetes and a previous myocardial infarction (MI). BACKGROUND: People with type 2 diabetes have an increased incidence of MI compared with the general population. Those with diabetes and MI have a worse prognosis than nondiabetic patients with cardiovascular disease. METHODS: The PROactive study was a prospective, multicenter, double-blind, placebo-controlled trial of 5,238 patients with type 2 diabetes and macrovascular disease. Patients were randomized to either pioglitazone or placebo in addition to their other glucose-lowering and cardiovascular medication. Treatment of diabetes, dyslipidemia, and hypertension was encouraged according to the International Diabetes Federation guidelines. Patients were followed for a mean of 2.85 years. The primary end point was the time to first occurrence of macrovascular events or death. Of the total cohort, the subgroup of patients who had a previous MI (n = 2,445 [46.7%]; n = 1,230 in the pioglitazone group and n = 1,215 in the placebo group) was evaluated using prespecified and post-hoc analyses. RESULTS: Pioglitazone had a statistically significant beneficial effect on the prespecified end point of fatal and nonfatal MI (28% risk reduction [RR]; p = 0.045) and acute coronary syndrome (ACS) (37% RR; p = 0.035). There was a 19% RR in the cardiac composite end point of nonfatal MI (excluding silent MI), coronary revascularization, ACS, and cardiac death (p = 0.033). The difference in the primary end point defined in the main PROactive study did not reach significance in the MI population (12% RR; p = 0.135). The rates of heart failure requiring hospitalization were 7.5% (92 of 1,230) with pioglitazone and 5.2% (63 of 1,215) with placebo. Fatal heart failure rates were similar (1.4% [17 of the 92] with pioglitazone versus 0.9% [11 of the 63] with placebo). CONCLUSIONS: In high-risk patients with type 2 diabetes and previous MI, pioglitazone significantly reduced the occurrence of fatal and nonfatal MI and ACS. (PROspective pioglitAzone Clinical Trial In macroVascular Events; http://www.clinicaltrials.gov/ct/show/NCT00174993?order = 1; ISRCTN NCT00174993). PMID- 17466232 TI - The relationship between neo-aortic root dilation, insufficiency, and reintervention following the Ross procedure in infants, children, and young adults. AB - OBJECTIVES: The purpose of this study was to describe the relationship between neo-aortic root size, neo-aortic insufficiency (AI), and reintervention at mid term follow-up. BACKGROUND: Data on neo-aortic valve function and growth after the Ross procedure in children are limited. METHODS: A total of 74 of 119 Ross patients from January 1995 to December 2003 had > or =2 follow-up echocardiograms at our institution and were included. Neo-aortic dimensions were converted to z scores and modeled over time. Kaplan-Meier analysis was used to assess freedom from neo-aortic outcomes, and predictors were identified through multivariate analysis. RESULTS: Median age at Ross was 9 years (range 3 days to 34 years). Over 4.7 years (range 3 months to 9.3 years) follow-up, there was disproportionate enlargement of the neo-aortic root (z-score increase of 0.75/year [p < 0.0001]). Neo-AI progressed > or =1 grade in 36% of patients and > or =2 grades in 15%. Nine patients (12%) had neo-aortic reintervention at 2.0 years (range 1.1 to 9.5 years) after the Ross procedure owing to severe neo-AI (n = 7), neo-aortic root dilation (n = 1), and neo-aortic pseudoaneurysm (n = 1). At 6 years after the Ross procedure, freedom from neo-aortic reintervention was 88%. Freedom from neo-aortic root z-score >4 was only 3% and from moderate or greater neo-AI was 60%. Longer follow-up time was associated with neo-aortic root dilation (p < 0.0001). Prior ventricular septal defect (VSD) repair predicted neo AI (p = 0.02) and reintervention (p = 0.03). Prior aortic valve replacement (p = 0.002) also predicted neo-AI. Neo-aortic root dilation was not associated with neo-AI or reintervention. CONCLUSIONS: At mid-term follow-up after the Ross procedure, neo-aortic root size increases significantly out of proportion to somatic growth, and neo-AI is progressive. Prior VSD repair and aortic valve replacement were associated with neo-AI and reintervention. PMID- 17466233 TI - Pacing-induced dyssynchrony during early reperfusion reduces infarct size. AB - OBJECTIVES: Considering the recent discovery of postconditioning, we investigated whether intermittent dyssynchrony immediately upon reperfusion induces cardioprotection as well. BACKGROUND: Intermittent dyssynchrony, induced by ventricular pacing, preconditions myocardium. METHODS: Isolated ejecting rabbit hearts were subjected to 30-min coronary occlusion and 2-h reperfusion. Control, left ventricular (LV) pacing preconditioning (LVPpreC) (3 x 5-min LV pacing), and LV pacing postconditioning (LVPpostC) (10 x 30-s LV pacing during early reperfusion) groups were studied. Mechanical effects of LV pacing were determined using local pressure-length loops (sonomicrometry), whereas effects on myocardial lactate release and coronary flow were assessed from coronary effluent and fluorescent microspheres, respectively. Anesthetized pigs underwent 60-min coronary occlusion and 3-h reperfusion in control and right ventricular (RV) pacing postconditioning groups (RVPpostC) (10 x 30-s RV pacing during early reperfusion). In all hearts, area at risk and infarct size were determined with blue dye and triphenyltetrazolium chloride staining, respectively. RESULTS: Infarct size, normalized to area at risk, was 47.0 +/- 12.3% in control rabbit hearts, but significantly smaller in LVPpreC (17.8 +/- 6.4%) and LVPpostC hearts (17.9 +/- 4.4%). Left ventricular pacing significantly altered regional mechanical work, but did not affect coronary flow or lactate release. In pigs, infarct size was significantly smaller in RVPpostC (9.8 +/- 3.0%) than in control (20.6 +/- 2.2%) animals. CONCLUSIONS: Intermittent dyssynchrony during early reperfusion reduces infarct size in 2 different animal models. Dyssynchrony induced postconditioning cannot be attributed to graded reperfusion but may be induced by modulation of local myocardial workload. Dyssynchrony-induced postconditioning opens new possibilities for cardioprotection in the clinical setting. PMID- 17466234 TI - President's page: On fulfilling our obligations to patients, the profession, and health care reform (excerpted from Dr. Dove's Speech Given at the 56th Convocation of the American College of Cardiology, March 26, 2007, New Orleans, Louisiana). PMID- 17466236 TI - The brain and the heart: independent or interactive? PMID- 17466237 TI - The anti-inflammatory properties of safflower oil and coconut oil may be mediated by their respective concentrations of vitamin E. PMID- 17466239 TI - Fresh-frozen osteochondral allograft reconstruction of a giant cell tumor of the talus. AB - The use of fresh-frozen osteochondral allografts has been reported for the treatment of talar fractures, osteochondral lesions, and tumors of the calcaneus. Currently, we are unaware of any reports in the literature addressing the use of fresh-frozen osteochondral allograft for the treatment of giant cell tumors in the talus. We report our attempt to eradicate an aggressive giant cell tumor of the talus while minimizing morbidity and loss of function via reconstruction with a fresh-frozen osteochondral allograft. This is the first report in the literature to propose such a treatment option for giant cell tumors in the talus. The patient was informed that a report of this case would be submitted for publication. PMID- 17466240 TI - Hallux valgus inheritance: pedigree research in 350 patients with bunion deformity. AB - Our objective was to construct 3-generation pedigree charts from 350 patients with hallux valgus. During a 1-year period, all consecutive patients (n = 1174) with a painful bunion deformity evaluated roentgenographically were asked to complete a detailed 3-generation family history questionnaire. We studied 350 probands (22 men, 328 women; male/female ratio, 1:14.9; mean age, 47.8 years). Juvenile hallux valgus was diagnosed in 15 patients. Three or more affected members were observed in pedigrees from 244 probands, 2 affected members in 71, and 1 affected member in 35 (proband) (affected subjects per pedigree ranged from 1 to 16). Ninety percent of probands had at least 1 family member affected. The hallux valgus penetrance according to pedigrees from all probands was 56%. The female sex predominated with regard to the gender of parents with hallux valgus, affected branch of the family, and gender of relatives with bunion deformity. Severity of hallux valgus was not significantly influenced by gender, the affected branch of the family, or gender of the affected relatives. Family history of bunion deformity was present in 90% of probands, with vertical transmission affecting some family members across 3 generations, which is compatible with autosomal dominant inheritance with incomplete penetrance. PMID- 17466241 TI - Spiral-shaped metallic implant in the treatment of Achilles tendon ruptures: an experimental study on the Achilles tendon of sheep. AB - There are 2 main factors contributing to the strength of tendon repair: the tensile strength of the material used in repair and the tendon-holding capacity of the suture configuration. In the current study, we aimed to find a technique with high repair strength by increasing both the tensile strength of the material and the tendon-holding capacity of the configuration. We developed metal implants (models 1 and 2) made from stainless-steel wire with 2 different spiral-shaped configurations. We measured tendon-holding capacities of these alternative implants biomechanically and compared them with frequently used suture techniques, the Bunnell and locking loop, which were achieved with 5 Ticron sutures. Sixty-four sheep Achilles' tendons were used in the study as 16 tendons in each group. Model 2 was more resistant to deformation under loading when compared with model 1. The results demonstrated that model 2 was superior to model 1 and both suture techniques. This study could be accepted as a step for reaching a strong tendon repair technique. It should be emphasized that the technique needs to be improved technically to make it convenient for clinical use. PMID- 17466242 TI - Comparison of a lateral hop test versus a forward hop test for functional evaluation of lateral ankle sprains. AB - The purposes of this study were to determine whether a lateral hop test was a more sensitive functional test over time than a forward hop test in assessing lateral ankle sprains, and whether lateral hop performance can predict a subjective score from an ankle rating scale. At the United States Military Academy, cadets presenting with ankle sprains during an 8-month period were included in this observational study. Patients were asked to perform a lateral hop for distance and a forward hop for distance on both the injured and uninjured lower extremities. The order of testing was randomized. After the hop trials, individuals completed a subjective questionnaire designed to assess functional ankle health. The lateral hop and subjective scores are components of the Sports Ankle Rating System. Patients were evaluated at the day of consent and at 1 week, 3 weeks, and 6 weeks. There were 29 patients, ages 18 to 22 years; 8 were women and 21 were men. A multivariable regression of analysis was performed to determine which subjective factors best predict the individual's subjective score. Although both the lateral and forward hop were statistically significant factors, neither was determined to be better than the other. PMID- 17466243 TI - Preliminary radiographic findings and sizing implications on patients undergoing bioabsorbable subtalar arthroereisis. AB - Metallic subtalar arthroereisis implants can require removal. Similarly configured bioabsorbable "interference screws" placed alternatively to metal implants may obviate removal. Radiographic imaging may show the location and sizing of the implant, and evaluate for implant degradation. Patients undergoing subtalar arthroereisis were evaluated with magnetic resonance imaging (computed tomography in one patient) to measure the size of the tarsal canal. The tarsal canal length, along with medial height and lateral height, were assessed. The patient's actual implant size was also noted as well as any signs of implant degradation, bony deformation, granulomas, and so forth. Six patients met the inclusion criteria. The range of implants used was 9 to 12 mm. The radiographic measurements of the tarsal canal were as follows: medial to lateral length, 12.8 +/- 3.4 mm; medial height, 7.3 +/- 2.5 mm; and lateral height, 8.0 +/- 1.7 mm. Two patients underwent implant removal. No cystic or degenerative changes were noted on plain radiographs with bioabsorbable implants. Bioabsorbable interference screws for subtalar arthroereisis placed in the tarsal canal may still require removal, although no detrimental changes were noted radiographically to the surrounding bony structures. The size of the current metallic implants on the market appears larger than the tarsal canal configuration. PMID- 17466244 TI - Medial and lateral malleolar arteries in ankle arthroscopy: a cadaver study. AB - Neurovascular injury may occur during ankle arthroscopy. The majority of complications are neurological injuries; however, vascular injuries do exist. Neurovascular structures are especially vulnerable during portal placement and debridement of anterior structures. Routine anteromedial and anterolateral portals are generally accepted to be safe; this is different from the anterocentral portal, which is associated with a higher risk of injury. However, injuries may occur in these relatively safe portals. The purpose of this cadaver study was to examine other relatively minor neurovascular structures such as medial and lateral malleolar arteries and to determine how these portals can be more safely placed. The distance between standard anteromedial, anterolateral portals and the medial and lateral malleolar arteries was measured in 18 ankles from 9 cadavers. These distances varied with the position of the ankle during portals placement, and measurements were obtained in both flexion and extension. The average distance in flexion and extension was 6.41 to 2.47 mm on the lateral side and 4.73 to 1.58 mm on the medial side. The distances significantly increased with ankle flexion and decreased with extension (P < .005). The current study demonstrated that there were other minor vascular structures at risk other than tibialis anterior artery and proper positioning of the ankle during portal placement, and that injury risk may be associated with ankle position. Ankle flexion may decrease the risk of damage to malleolar arteries and decrease minor vascular complications such as postoperative bleeding and hematoma. PMID- 17466245 TI - Isolated floating first metatarsal: report of an unusual injury. AB - The floating metatarsal is an extremely rare type of injury in which the first metatarsal is dislocated both proximally and distally. Associated injuries and especially metatarsal fracture appear to be rules rather than exceptions. An unreported type of foot injury consisting of isolated floating first metatarsal is described. The absence of any metatarsal fracture makes it exceptional. The importance of following an order of reduction of the joints to relieve tension on the plantar fascia has been highlighted. When examining a patient with metatarsophalangeal joint injury, one should always look for injury at the tarsometatarsal joint and vice versa, because various concomitant injuries are possible and misdiagnose can cause long-term secondary disability. PMID- 17466246 TI - Concomitant talar neck fracture and Achilles tendon rupture. AB - A 56-year-old man fell down 1 m from a ladder and sustained a forced dorsiflexion injury to his right ankle when his foot contacted a lower rung, which resulted in the rare combination of a Hawkins II fracture of the neck of the talus and a concomitant rupture of the Achilles tendon. Clinical examination and diagnostic imaging confirmed the injuries, and surgical fixation of the fracture and repair of the Achilles tendon were achieved by means of a posterior approach. Healing proceeded unremarkably, and, at 18 months postoperatively, the patient had regained full function despite a 5 degrees limitation of subtalar joint range of motion. PMID- 17466247 TI - Arthroscopic treatment of synovial chondromatosis of the ankle. AB - We report a case of synovial chondromatosis of the ankle joint, which has been successfully treated with arthroscopic removal of loose bodies and synovectomy, with the patient immediately returning to school and activities of daily living. Recovery after arthroscopic debridement and loose body removal is much shorter in comparison with arthrotomy, and there is no need of immobilization postoperatively. PMID- 17466248 TI - Merkel cell carcinoma of the foot: case report and review of the literature. AB - Merkel cell carcinomas are rare neuroendocrine tumors of the skin with highly malignant behavior. These tumors arise from Merkel cells, which are slowly adapting type 1 mechanoreceptors that occur in hair follicles and the basal layer of the epidermis. Merkel cell carcinomas are typically located in the dermis and frequently extend into the subcutaneous fat and lymphatics. This case report describes the magnetic resonance imaging and pathologic findings, surgical treatment, and postoperative results with review of the literature for Merkel cell carcinoma involving the foot of a 72-year-old man with a 2-month history of a painless, enlarging mass that interfered with his ability to wear a shoe. PMID- 17466249 TI - Tarsal tunnel decompression in leg lengthening and deformity correction of the foot and ankle. AB - Decompression of the tarsal tunnel has been performed since the 1960s and has resulted in variable outcomes. The success rate of this procedure has ranged from 44% to 95%. We review the anatomy of the tarsal tunnel region and offer a detailed surgical approach for the decompression of its contents. We also discuss the role of this procedure in leg lengthening and deformity correction of the foot and ankle. PMID- 17466250 TI - An eight-toed foot: a rare pedal polydactyly. AB - Polydactyly is a fairly common congenital condition of the foot and is characterized literally by supernumerary toes (digit or metatarsal). The frequency of polydactyly varies widely among populations. It may be an isolated condition or part of a congenital syndrome. Eight-toed polydactyly is a distinctly rare congenital foot anomaly. In this report, an 18-month-old child with pre-axial 8-toed right foot had been treated with excision of the excess rays with muscular, tendentious, and ligamentous reconstructions. The patient had presented a good postoperative result. Through monthly follow-up visits for 1 year, the parents expressed excellent cosmesis, shoe fit, and walking function. PMID- 17466251 TI - IT in health care: sociotechnical approaches "To Err is System". PMID- 17466252 TI - Object memory and change detection: dissociation as a function of visual and conceptual similarity. AB - People often fail to detect a change between two visual scenes, a phenomenon referred to as change blindness. This study investigates how a post-change object's similarity to the pre-change object influences memory of the pre-change object and affects change detection. The results of Experiment 1 showed that similarity lowered detection sensitivity but did not affect the speed of identifying the pre-change object, suggesting that similarity between the pre- and post-change objects does not degrade the pre-change representation. Identification speed for the pre-change object was faster than naming the new object regardless of detection accuracy. Similarity also decreased detection sensitivity in Experiment 2 but improved the recognition of the pre-change object under both correct detection and detection failure. The similarity effect on recognition was greatly reduced when 20% of each pre-change stimulus was masked by random dots in Experiment 3. Together the results suggest that the level of pre-change representation under detection failure is equivalent to the level under correct detection and that the pre-change representation is almost complete. Similarity lowers detection sensitivity but improves explicit access in recognition. Dissociation arises between recognition and change detection as the two judgments rely on the match-to-mismatch signal and mismatch-to-match signal, respectively. PMID- 17466253 TI - Chlorfenapyr: a pyrrole insecticide for the control of pyrethroid or DDT resistant Anopheles gambiae (Diptera: Culicidae) mosquitoes. AB - Owing to the development and spread of pyrethroid resistance in Anopheles gambiae in Africa there is an urgent need to develop alternative insecticides to supplement the pyrethroids. Chlorfenapyr is a pyrrole insecticide first commercialized for the control of agricultural pests and termites. Performance against An. gambiae bearing kdr (pyrethroid and DDT resistance) or Ace-1(R) insensitive acetylcholinesterase (organophosphate and carbamate resistance) mechanisms was studied using a variety of adult bioassay tests including a simulated-experimental hut system (tunnel tests) that allows uninhibited mosquito behaviour/insecticide interactions. Strains resistant to pyrethroids and organophosphates showed no cross resistance to chlorfenapyr. In cone bioassays on treated netting the mortality of adult mosquitoes showed an unexpected curvilinear response, with highest mortality occurring at intermediate dosages. Adults expressed irritability to chlorfenapyr at higher dosages, which might explain the dosage-mortality trend. Toxic activity of chlorfenapyr was slow compared to conventional neurotoxic insecticides and additional mortality occurred between 24h and 72 h. In tunnel tests, the dosage-mortality trend showed a more typical sigmoid response and most mortality occurred during the first 24h. Mosquito penetration through the holed, treated netting showed only limited inhibition and blood-feeding was not inhibited. Mortality rates in the kdr strain exposed to chlorfenapyr treated netting in tunnel tests were much higher than with permethrin treated netting over the same 100-500 mg/m(2) dosage range. Chlorfenapyr has potential for malaria control in treated-net or residual spraying applications in areas where mosquitoes are pyrethroid resistant. For treated-net applications chlorfenapyr might be combined with pyrethroid as a mixture to provide personal protection as well as to give control of resistant mosquitoes. PMID- 17466254 TI - Flight decks and free flight: where are the system boundaries? AB - The change from managed to free flight is expected to have large effects, over and above the intended efficiency gains. Human factor concerns have understandably focused on how free flight may affect the pilots in the cockpit. Yet it is necessary to see the change from managed to free flight as more than just an increment to the pilots' work. Despite the best intentions the transition will not be a case of a smooth, carefully planned and therefore uneventful introduction of a new technology. It is more likely to be a substantial change to an already challenging working environment, in the air as well as on the ground. The significant effects will therefore not just happen within the existing structure or distribution of work and responsibilities, but affect the structure of work itself. This paper takes a look at free flight from a cognitive systems engineering perspective and identifies two major concerns: first what effects free flight has on the boundaries of the joint cognitive systems, and second how this affects demands to control. The conclusion is that both will change considerably and that we need to understand the nature of these changes before focusing on the possible effects of free flight on pilots' performance. PMID- 17466255 TI - Isolation, purification and characterization of histidino-threosidine, a novel Maillard reaction protein crosslink from threose, lysine and histidine. AB - We isolated a novel acid-labile yellow chromophore from the incubation of lysine, histidine and d-threose and identified its chemical structure by one and two dimensional NMR spectroscopy combined with LC-tandem mass spectrometry. This new cross-link exhibits a UV absorbance maximum at 305 nm and a molecular mass of 451 Da. The proposed structure is 2-amino-5-(3-((4-(2-amino-2-carboxyethyl)-1H imidazol-1-yl)methyl)-4-(1,2-dihydroxyethyl)-2-formyl-1H-pyrrol-1-yl)pentatonic acid, a cross-link between lysine and histidine with addition of two threose molecules. It was in part deduced and confirmed through synthesis of the analogous compound from n-butylamine, imidazole and d-threose. We assigned the compound the trivial name histidino-threosidine. Systemic incubation revealed that histidino-threosidine can be formed in low amounts from fructose, glyceraldehyde, methylglyoxal, glycolaldehyde, ascorbic acid, and dehydroascorbic acid, but at a much higher yield with degradation products of ascorbic acid, i.e. threose, erythrose, and erythrulose. Bovine lens protein incubated with 10 and 50 mM threose for two weeks yielded 560 and 2840 pmol/mg histidino-threosidine. Histidino-threosidine is to our knowledge the first Maillard reaction product known to involve histidine in a crosslink. PMID- 17466256 TI - Apoptosis and inhibition of gap-junctional intercellular communication induced by LA-12, a novel hydrophobic platinum(IV) complex. AB - A new hydrophobic platinum(IV) complex, LA-12, a very efficient anticancer drug lacking cross-resistance with cisplatin (CDDP), is now being tested in clinical trials. Here we investigated the apoptogenic activity of LA-12 and its effect on gap-junctional intercellular communication (GJIC) in the rat liver epithelial cell line WB-F344. LA-12 induced apoptosis much more efficiently than did CDDP due to a combination of rapid penetration into the cell and attack on DNA, leading to fast activation of p53 and caspase-3. Exposure of WB-F344 cells to LA 12 led to rapid induction of the time- and dose-dependent decrease in GJIC. On the molecular level, loss of GJIC induced by LA-12 was mediated by activation of extracellular signal-regulated kinase (ERK)-1 and ERK-2, as demonstrated by the use of inhibitors of ERK activation. Inhibition of GJIC was linked to rapid hyperphosphorylation of connexin-43 and disappearance of connexon clusters from membranes, which was not observed in the case of CDDP. PMID- 17466257 TI - Structural and energetic aspects of Grb2-SH2 domain-swapping. AB - The SH2 domain of growth factor receptor-bound protein 2 (Grb2) has been the focus of numerous studies, primarily because of the important roles it plays in signal transduction. More recently, it has emerged as a useful protein to study the consequences of ligand preorganization upon energetics and structure in protein-ligand interactions. The Grb2-SH2 domain is known to form a domain swapped dimer, and as part of our investigations toward correlating structure and energetics in biological systems, we examined the effects that domain-swapping dimerization of the Grb2-SH2 domain had upon ligand binding affinities. Isothermal titration calorimetry was performed using Grb2-SH2 in both its monomeric and domain-swapped dimeric forms and a phosphorylated tripeptide AcNH pTyr-Val-Asn-NH(2) that is similar to the Shc sequence recognized by Grb2-SH2 in vivo. The two binding sites of domain-swapped dimer exhibited a 4- and a 13-fold reduction in ligand affinity compared to monomer. Crystal structures of peptide bound and uncomplexed forms of Grb2-SH2 domain-swapped dimer were obtained and reveal that the orientation of residues V122, V123, and R142 may influence the conformation of W121, an amino acid that is believed to play an important role in Grb2-SH2 ligand sequence specificity. These findings suggest that domain-swapping of Grb2-SH2 not only results in a lower affinity for a Shc-derived ligand, but it may also affect ligand specificity. PMID- 17466258 TI - Does the inhibition of c-myc expression mediate the anti-tumor activity of PPAR's ligands in prostate cancer cell lines? AB - Peroxisome proliferator-activated receptor gamma (PPARgamma) ligands seem to induce anticancer effects on prostate cancer cells, but the mechanism is not clear. The effect of PPARgamma ligands omega-6 fatty acids and ciglitazone (2-15 microM)--on proliferation, and apoptosis of LNCaP, PC-3, DU145, CA-K and BPH-K cells was studied. PPARgamma ligands led to: (1) reduction of proliferation (20 50%) of all the studied cell lines, (2) stimulation of differentiation of prostate cancer cells through an increased expression (1.5-3-fold: LNCaP, DU145, BPH-K) or reexpression (PC-3, CA-K) of E-cadherin with parallel inhibition of N cadherin expression (PC-3, CA-K) and (3) down-regulation (1-2-fold) of beta catenin and c-myc expression. The selective PPARgamma antagonist GW9662 abolished the effect of those ligands on prostate cancer cells. These results suggest that inhibition of beta-catenin and in effect c-myc expression through activation of PPARgamma may help prostate cancer cells to restore several characteristics of normal prostate cells phenotype. PMID- 17466259 TI - Genotypic diversity of Streptococcus mutans in 3- to 4-year-old Chinese nursery children suggests horizontal transmission. AB - OBJECTIVE: The aims of this study were to investigate the colonisation of Streptococcus mutans and to determine the possibility of horizontal transmission in Chinese nursery children. DESIGN: Fifty-six children aged between 3 and 4 years old from a nursery were included in this study. Twenty-four children were from the day and night nursery class, the others were from the day nursery class. Dental plaque samples were collected with sterile toothpicks and cultured on S. mutans-selective tryptone-yeast-cystine agar supplemented with 0.2U/ml bacitracin and 15% sucrose. The typical isolates of S. mutans were identified by classical microbiological methods and genotyped by arbitrarily primed PCR (AP-PCR) and restriction fragment length polymorphism analysis. RESULTS: S. mutans was isolated from 41 of the 56 children. The prevalence of S. mutans was higher in the children from day nursery class, compared with the children from day and night nursery class (P<0.05). A total of 140 S. mutans isolates from 41 children were analysed by AP-PCR, and 88 different amplitypes were identified. Of 41 children with S. mutans isolates, 82.9% carried two or more genotypes. Two genotypes of S. mutans were repeatedly isolated among 13 children in the day and night nursery class, and one genotype was isolated from two children in the day nursery class. CONCLUSION: The presence of matching genotypes of S. mutans among children attending the same nursery suggests the possibility of horizontal transmission. PMID- 17466260 TI - The inside pH determines rates of electron and proton transfer in vesicle reconstituted cytochrome c oxidase. AB - Cytochrome c oxidase is the terminal enzyme in the respiratory chains of mitochondria and many bacteria where it translocates protons across a membrane thereby maintaining an electrochemical proton gradient. Results from earlier studies on detergent-solubilized cytochrome c oxidase have shown that individual reaction steps associated with proton pumping display pH-dependent kinetics. Here, we investigated the effect of pH on the kinetics of these reaction steps with membrane-reconstituted cytochrome c oxidase such that the pH was adjusted to different values on the inside and outside of the membrane. The results show that the pH on the inside of the membrane fully determines the kinetics of internal electron transfers that are linked to proton pumping. Thus, even though proton release is rate limiting for these reaction steps (Salomonsson et al., Proc. Natl. Acad. Sci. USA, 2005, 102, 17624), the transition kinetics is insensitive to the outside pH (in the range 6-9.5). PMID- 17466261 TI - Transport and transporters in the endoplasmic reticulum. AB - Enzyme activities localized in the luminal compartment of the endoplasmic reticulum are integrated into the cellular metabolism by transmembrane fluxes of their substrates, products and/or cofactors. Most compounds involved are bulky, polar or even charged; hence, they cannot be expected to diffuse through lipid bilayers. Accordingly, transport processes investigated so far have been found protein-mediated. The selective and often rate-limiting transport processes greatly influence the activity, kinetic features and substrate specificity of the corresponding luminal enzymes. Therefore, the phenomenological characterization of endoplasmic reticulum transport contributes largely to the understanding of the metabolic functions of this organelle. Attempts to identify the transporter proteins have only been successful in a few cases, but recent development in molecular biology promises a better progress in this field. PMID- 17466262 TI - Sex differences in emotional and physiological responses to the Trier Social Stress Test. AB - Women are more likely than men to be diagnosed with depression and anxiety related disorders, and it has been hypothesized that this difference is related to sex differences in stress reactivity. Women typically report higher levels of negative affect than men in response to psychosocial stressors, but the evidence for sex differences in physiological reactivity to stressful situations is not consistent. The present study sought to expand this work by evaluating sex differences in reactivity to a social stress challenge across neuroendocrine, autonomic and affective response domains. Participants (32 women, 30 men) completed a standardized psychosocial stress challenge (i.e., the Trier Social Stress Test (TSST)), during which several physiological (e.g., cortisol reactivity, heart rate) and psychological (e.g., depression, irritability, anger, fear) measures were assessed. The findings demonstrated that cortisol reactivity and the magnitude of autonomic responding failed to reliably discriminate between women and men. However, women reported more fear, irritability, confusion and less happiness immediately following the TSST compared to men. The broader implications of these results and how they relate to sex differences in the etiology and clinical presentation of anxiety and mood disorders are discussed. PMID- 17466263 TI - Not just right experience: is it influenced by feelings of guilt? AB - Individuals with obsessive-compulsive disorder (OCD) experience increased guilt. Further, these individuals often report uncomfortable sensations of things being not quite right ("not just right experiences"--NJREs). As to the relation between these psychological phenomena, it was hypothesized that feelings of guilt may enhance NJRE. In two experiments, we demonstrated that the induction of a guilty emotion resulted in increased NJRE, and this finding was qualified by an interaction with trait guilt. Induced guilt was followed by stronger feelings of things being not just right only in high-trait-guilt participants. In the low trait-guilt participants NJRE was weaker. Moreover, we found a meaningful relationship between both NJRE and trait guilt and OCD features. PMID- 17466264 TI - Constitutive GABA expression via a recombinant adeno-associated virus consistently attenuates neuropathic pain. AB - Peripheral neuropathic pain is a common clinical problem with few existing treatments. Previously, we constructed rAAV bearing GAD65 and demonstrated that GAD65 and GABA can be constitutively produced in the CNS. To investigate the beneficial effects of GAD65 produced by rAAV and resulting GABA release in peripheral neuropathic pain, we established a neuropathic pain rat model. The direct administration of rAAV-GAD65 to dorsal root ganglion induced constitutive GAD65 expression, which was readily detected by immunohistochemistry. Both allodynic and hyperalgeic behavior tests suggested that neuropathic pain was noticeably reduced, along with the transgenic GAD65 expression. Moreover, the magnitude of pain relief was maintained during the entire experimental period. Concomitantly, the significant enhancement in GABA release following transgenic GAD65 expression was identified in vivo. Taken all together, these results provide evidence that persistent GAD65 and subsequent GABA expression in DRGs via rAAV effectively attenuates peripheral neuropathic pain for long period of time. PMID- 17466265 TI - Phospholipid binding properties and functional characterization of a sea urchin phospholipase Cdelta in urchin and mouse eggs. AB - We recently identified a novel phospholipase Cdelta isoform, PLC-deltasu, in sea urchin gametes, whose precise functional role during fertilization and early embryogenesis remains unknown. Here, we characterized the binding of the PLC deltasu PH domain to different phosphatidylinositol (PI) phospholipids and studied changes in its localization during fertilization. The PLC-deltasu PH domain bound most strongly to PI(3,4)P(2) and PI(3,5)P(2) phospholipids, in contrast to the PLCdelta1 PH domain which bound predominantly to PI(4,5)P(2). A green fluorescent protein tagged PLC-deltasu PH domain localized to the plasma membrane and its localization increased at fertilization and following addition of a Ca(2+) ionophore. However, recombinant PLC-deltasu failed to cause Ca(2+) signals like those seen at fertilization, in mouse and sea urchin eggs. Our findings suggest that PLC-deltasu is unlikely to be directly involved in the process of egg activation but may play a role in mediating extracellular signals transmitted via the PI 3'-kinase pathway. PMID- 17466266 TI - Efficient conversion of ES cells into myogenic lineage using the gene-inducible system. AB - We established genetically engineered ES (ZHTc6-MyoD) cells that harbor a tetracycline-regulated expression vector encoding myogenic transcriptional factor MyoD, for the therapy of muscle diseases, especially Duchenne muscular dystrophy (DMD). Almost all the ZHTc6-MyoD cells were induced into muscle lineage after removal of tetracycline. The undifferentiated ZHTc6-MyoD cells are Sca-1+ and c kit+, but CD34-, all well-known markers for mouse hematopoietic stem cells. In addition, they are able to maintain themselves in the undifferentiated state, even after one month of culture. Therefore, it is possible to obtain a large quantity of ZHTc6-MyoD cells in the undifferentiated state that maintain the potential to differentiate only into muscle lineage. Additionally, at two weeks post-injection of these cells into muscle of mdx, a model mouse of DMD, clusters of dystrophin-positive myofibers were observed at the injection site. Therefore, ES cells have considerable therapeutic potential for treating muscle diseases. PMID- 17466267 TI - A critical role of vascular endothelial growth factor D in zebrafish embryonic vasculogenesis and angiogenesis. AB - The VEGF family comprises seven members that are designated VEGF-A, VEGF-B, VEGF C, VEGF-D, VEGF-E, placental growth factor (PlGF), and VEGF-F. Of these factors, VEGF-D plays important roles for angiogenesis and lymphangiogenesis, and could promote tumor growth and lymphatic metastasis. In this study, we identified a zebrafish VEGF-D homolog that encodes a 272 amino acid protein including a PDGF (platelet-derived growth factor) domain characteristic to VEGF family. Expression profile demonstrated that the VEGF-D began expressed from 13 somite stage. Microinjecting zVEGF-D mRNA into zebrafish 1-cell stage embryos resulted in severe misguidance of intersegmental vessels (ISV) and abnormal connection between dorsal aorta and caudal vein. Microangiography indicated that these abnormal ISVs were not functional. Our studies therefore identified the first non mammalian VEGF-D and established its in vivo role for vascular system development during vertebrate embryogenesis and provided an alternative animal model to further reveal functions of VEGF-D. PMID- 17466268 TI - Signaling pathways of the early differentiation of neural stem cells by neurotrophin-3. AB - Neurotrophin-3 (NT-3) is well known to play an important role in facilitating neuronal survival and differentiation during development. However, the mechanisms by which neurotrophin-3 promotes prolonged Akt/MAPK signaling at an early stage are not well understood. Here, we report that NT-3 works at an early stage of neuronal differentiation in mouse neural stem cells (NSCs). After treatment with NT-3 for 12h, more NSCs differentiated into neurons than did untreated cells. These findings demonstrated that stimulation with NT-3 causes NSCs to differentiate into neurons through a phosphatidylinositol 3-kinase (PI3K)/Akt signaling pathway and the phosphorylated extracellular signal-regulated kinase (ERK) pathway. In addition, treatment with NT-3 induced neurite outgrowth by specific phosphorylation of p38 MAPK, which was accompanied by neuronal differentiation. Taken together, these results suggest that NT-3, along with the Trk C receptors in NSCs, might lead to the survival and neuronal differentiation of NSCs via two distinct downstream signaling pathways at an early stage of neuronal differentiation. PMID- 17466269 TI - Direct effects of apelin on cardiomyocyte contractility and electrophysiology. AB - Apelin, the ligand for the angiotensin receptor like-1, has been implicated in the pathogenesis of atrial fibrillation and heart failure. However, it is unknown if apelin has direct effects on cardiomyocyte contractility and electrophysiology. APJ-like immunoreactivity was localized to T-tubules and intercalated disc area in isolated adult rat ventricular myocytes. Apelin (1 nM) significantly increased sarcomere shortening in normal as well as failing cardiomyocytes. The transient increase in shortening was not accompanied by increased [Ca(2+)] transient amplitude. Apelin significantly activated the sarcolemmal Na(+)/H(+) exchanger (NHE) and increased intracellular pH. Moreover, apelin (10 nM) increased conduction velocity in monolayers of cultured neonatal rat cardiac myocytes. Our results demonstrate for the first time that apelin has direct effects on the propagation of action potential and contractility in cardiomyocytes. One of the mechanisms involved in the inotropic effect may be an increased myofilament sensitivity to Ca(2+) as apelin enhanced the activity of NHE with consequent intracellular alkalinization. PMID- 17466270 TI - ATP promotes NCX-reversal in aortic smooth muscle cells by DAG-activated Na+ entry. AB - Reversal of the plasma membrane Na(+)/Ca(2+) exchanger (NCX) has been shown to mediate Ca(2+) influx in response to activation of G-protein linked receptors. Functional coupling of reverse-mode NCX with canonical transient receptor potential channels (TRPC), specifically TRPC6, has recently been demonstrated by our laboratory to mediate Ca(2+) influx in rat aortic smooth muscle cells (RASMCs) following ATP stimulation. In this communication, we provide further detail of this functional coupling by indirectly measuring NCX reversal. We found that NCX reversal, induced by the removal of extracellular Na(+), was increased following stimulation with ATP and the diacylglycerol analog 1-Oleoyl-2-acetyl-sn glycerol. This increased NCX reversal was attenuated by SKF-96365, an inhibitor of non-selective cation channels, and by activation of protein kinase C with phorbol ester 12-tetradecanoylphorbol-13 acetate. These data are consistent with the known properties of TRPC6 and further support that functional coupling of TRPC6 and NCX occurs via a receptor-operated, rather than store-operated, cascade in RASMCs. PMID- 17466271 TI - Celecoxib induces apoptosis by inhibiting the expression of survivin in HeLa cells. AB - The effect of celecoxib, a cyclooxygenase-2 selective inhibitor, on a human cervical cancer cell line, HeLa cells, was examined. We found that celecoxib increased DNA ladder formation and the activity of caspase-3, indicating that celecoxib induced apoptosis in HeLa cells. Celecoxib suppressed the expression of an anti-apoptotic protein, survivin, in both protein and mRNA levels. The overexpression of survivin overrode caspase-3 activation induced by celecoxib. Subsequently, we performed luciferase reporter assay with the reporter vector containing human survivin promoter region and electrophoretic mobility shift assay and found that the -75 to -66 bp region relative to the initiating codon played an important role in celecoxib action to suppress survivin promoter activity. Our findings might provide a new insight into the anti-cancer effects of celecoxib. PMID- 17466272 TI - Formation of human hepatocyte-like cells with different cellular phenotypes by human umbilical cord blood-derived cells in the human-rat chimeras. AB - We took advantage of the proliferative and permissive environment of the developing pre-immune fetus to develop a noninjury human-rat xenograft small animal model, in which the in utero transplantation of low-density mononuclear cells (MNCs) from human umbilical cord blood (hUCB) into fetal rats at 9-11 days of gestation led to the formation of human hepatocyte-like cells (hHLCs) with different cellular phenotypes, as revealed by positive immunostaining for human specific alpha-fetoprotein (AFP), cytokeratin 19 (CK19), cytokeratin 8 (CK8), cytokeratin 18 (CK18), and albumin (Alb), and with some animals exhibiting levels as high as 10.7% of donor-derived human cells in the recipient liver. More interestingly, donor-derived human cells stained positively for CD34 and CD45 in the liver of 2-month-old rat. Human hepatic differentiation appeared to partially follow the process of hepatic ontogeny, as evidenced by the expression of AFP gene at an early stage and albumin gene at a later stage. Human hepatocytes generated in this model retained functional properties of normal hepatocytes. In this xenogeneic system, the engrafted donor-derived human cells persisted in the recipient liver for at least 6 months after birth. Taken together, these findings suggest that the donor-derived human cells with different cellular phenotypes are found in the recipient liver and hHLCs hold biological activity. This humanized small animal model, which offers an in vivo environment more closely resembling the situations in human, provides an invaluable approach for in vivo investigating human stem cell behaviors, and further in vivo examining fundamental mechanisms controlling human stem cell fates in the future. PMID- 17466273 TI - Hsp70 is a new target of Sgt1--an interaction modulated by S100A6. AB - In this work, we identified Hsp70 as a novel target of the Sgt1 protein. Using co immunoprecipitation, affinity chromatography and ELISA we showed that, besides Hsp90, Sgt1 interacts with the heat shock protein, Hsp70. We also found that a deletion mutant of Sgt1, devoid of the C-terminal region, did not bind to either Hsp70 or Hsp90 proteins. Overexpression of S100A6, a calcium binding protein that interacts with the C-terminal part of Sgt1, decreased the amount of chaperone bound to Sgt1. However, the effect of S100A6 on this interaction was not observed in BAPTA/AM treated cells in which Ca(2+) level was decreased. This suggests that the interaction of Sgt1 with Hsp70 and Hsp90 is regulated by S100A6 in a Ca(2+) dependent manner. PMID- 17466274 TI - Search of type 2 diabetes susceptibility gene on chromosome 20q. AB - Significant evidence of linkage to type 2 diabetes (T2D) has been shown in a relatively broad region on chromosome 20q, where the hepatocyte nuclear factor 4alpha (HNF4A) has been noted as a positional candidate. To systematically evaluate genetic susceptibility to T2D in the relevant region, we examined the disease association by using 1145 SNPs in two-step screening in the Japanese population. The marker screening enabled us to identify significant disease association in the lipopolysaccharide binding protein (LBP) but not in the HNF4A locus. In a 17.7-Mb interval screened, the strongest association was identified for a SNP, rs2232592, located in the intron of LBP, with an estimated odds ratio of 1.73 (95% CI 1.30-2.31) (P=0.0002) in the whole study panel involving 675 case and 474 control subjects. Our data suggest that the LBP gene may confer genetic susceptibility to T2D and this warrants further replication study. PMID- 17466275 TI - Biogenesis and processing of the amyloid precursor protein in the early secretory pathway. AB - The beta-amyloid peptide is an aggregation-prone peptide that is released from the amyloid precursor protein (APP) after cleavage by the beta- and gamma secretase. A number of studies have suggested that processing of APP by beta- and gamma-secretase occurs not only at the cell surface and in the endosomal compartments but also in the endoplasmic reticulum (ER) and Golgi complex. Here, we studied the role of the early secretory pathway in processing of APP. For this purpose, APP was in vitro translated in semi-permeabilized cells, which have a functionally intact ER and Golgi complex but lack a functional plasma membrane. We show that the beta-secretase cleavage product C99 is generated in the early secretory pathway. Moreover, nicastrin and presenilin, two components of the gamma-secretase complex, interacted with newly synthesized APP. Administration of the gamma-secretase inhibitor L685,458 caused accumulation of full length APP and C99. Full length APP but not C99 interacted with several protein quality control ER chaperones including the thiol oxidoreductase ERp57. Our in vitro study suggests that newly synthesized APP is subject to amyloidogenic processing during the initial phases of the secretory pathway. PMID- 17466276 TI - Cilengitide inhibits proliferation and differentiation of human endothelial progenitor cells in vitro. AB - Bone marrow derived hematopoietic stem cells can function as endothelial progenitor cells. They are recruited to malignant tumors and differentiate into endothelial cells. This mechanism of neovascularization termed vasculogenesis is distinct from proliferation of pre-existing vessels. To better understand vasculogenesis we developed a cell culture model with expansion and subsequent endothelial differentiation of human CD133(+) progenitor cells in vitro. alpha(v)beta(3)-integrins are expressed by endothelial cells and play a role in the attachment of endothelial cells to the extracellular matrix. We investigated the effect of Cilengitide, a peptide-like, high affinity inhibitor of alpha(v)beta(3)- and alpha(v)beta(5)-integrins in our in vitro system. We could show expression of alpha(v)beta(3)-integrin on 60+/-9% of non-adherent endothelial progenitors and on 91+/-7% of differentiated endothelial cells. alpha(v)beta(3)-integrin was absent on CD133(+) hematopoietic stem cells. Cilengitide inhibited proliferation of CD133(+) cells in a dose-dependent manner. The development of adherent endothelial cells from expanded CD133(+) cells was reduced even stronger by Cilengitide underlining its effect on integrin mediated cell adhesion. Expression of endothelial antigens CD144 and von Willebrand factor on differentiating endothelial precursors was decreased by Cilengitide. In summary, Cilengitide inhibits proliferation and differentiation of human endothelial precursor cells underlining its anti-angiogenic effects. PMID- 17466277 TI - Activity of piperaquine and other 4-aminoquinoline antiplasmodial drugs against chloroquine-sensitive and resistant blood-stages of Plasmodium falciparum. Role of beta-haematin inhibition and drug concentration in vacuolar water- and lipid phases. AB - Chloroquine (CQ), a 4-aminoquinoline, accumulates in acidic digestive vacuoles of the malaria parasite, preventing conversion of toxic haematin to beta-haematin. We examine how bis 4-aminoquinoline piperaquine (PQ) and its hydroxy-modification (OH-PQ) retain potency on chloroquine-resistant (CQ-R) Plasmodium falciparum. For CQ, PQ, OH-PQ and 4 and 5, representing halves of PQ, beta-haematin inhibitory activity (BHIA) was assayed, while potency was determined in CQ-sensitive (CQ-S) and CQ-R P. falciparum. From measured pK(a)s and the pH-modulated distribution of base between water and lipid (logD), the vacuolar accumulation ratio (VAR) of charged drug from plasma water (pH 7.4) into vacuolar water (pH 4.8) and lipid accumulation ratio (LAR) were calculated. All agents were active in BHIA. In CQ S, PQ, OH-PQ and CQ were equally potent while 4 and 5 were 100 times less potent. CQ with two basic centres has a VAR of 143,482, while 4 and 5, with two basic centres of lower pK(a)s have VARs of 1287 and 1966. In contrast PQ and OH-PQ have four basic centres and achieve VARs of 104,378 and 19,874. This confirms the importance of VAR for potency against CQ-S parasites. Contrasting results were seen in CQ-R. 5, PQ and OH-PQ with LARs of 693; 973,492 and 398,118 (compared with 8.25 for CQ) showed similar potency in CQ-S and CQ-R. Importance of LAR for potency against CQ-R parasites probably reflects ability to block efflux by hydrophobic interaction with PfCRT but may relate to beta-haematin inhibition in vacuolar lipid. PMID- 17466278 TI - Accumulation, platinum-DNA adduct formation and cytotoxicity of cisplatin, oxaliplatin and satraplatin in sensitive and resistant human osteosarcoma cell lines, characterized by p53 wild-type status. AB - P53 gene status is implicated in the cytotoxic drug sensitivity and published research has been mostly addressed to cisplatin (CDDP) activity. Previous study in our laboratory considered p53 mutant cell lines A431 (parental) and A431/Pt (CDDP-resistant counterpart, resistance factor R.F.=2.6). For a comparison which contributes to a deeper appreciation of the process that mediates the Pt drug cellular effects, we extended our investigation to the p53 wild-type cell lines U2-OS (human osteosarcoma) and its CDDP-resistant counterpart U2-OS/Pt (R.F.=5). We compared the activity of CDDP, oxaliplatin (L-OHP) and satraplatin (JM216) whose hydrophobicity rank is JM216>L-OHP>CDDP. In U2-OS cells the three drugs accumulated similarly, while in U2-OS/Pt the most hydrophobic drugs were privileged. No significant differences in efflux were observed between sensitive and resistant cell lines. The growing of CDDP resistance seems to be overcome by increasing the hydrophobicity of the Pt agent. An almost linear trend seems to relate R.F. and drug hydrophobicity in U2-OS/Pt and A431/Pt cells. DNA platination in U2-OS as in A431 cells is at the lowest levels for L-OHP. In U2-OS cell line the IC(50) of CDDP (17.6 microM) and JM216 (88.02 microM) do not correlate with their similar levels of Pt-DNA adducts (mean value approximately 0.14 pmol Pt/microg DNA). The presence of a wild-type p53 exalts either CDDP cytotoxicity (two-fold more active in U2-OS than in A431 cells) and CDDP resistance in comparison to a p53 mutant type. The p53 status seems to not improve JM216 or L-OHP cytotoxicity in both cell lines. PMID- 17466279 TI - Procedure of rectal temperature measurement affects brain, muscle, skin, and body temperatures and modulates the effects of intravenous cocaine. AB - Rectal probe thermometry is commonly used to measure body core temperature in rodents because of its ease of use. Although previous studies suggest that rectal measurement is stressful and results in long-lasting elevations in body temperatures, we evaluated how this procedure affects brain, muscle, skin, and core temperatures measured with chronically implanted thermocouple electrodes in rats. Our data suggest that the procedure of rectal measurement results in powerful locomotor activation, rapid and strong increases in brain, muscle, and deep body temperatures, as well as a biphasic, down-up fluctuation in skin temperature, matching the response pattern observed during tail-pinch, a representative stressful procedure. This response, moreover, did not habituate after repeated day-to-day testing. Repeated rectal probe insertions also modified temperature responses induced by intravenous cocaine. Under quiet resting conditions, cocaine moderately increased brain, muscle, and deep body temperatures. However, during repeated rectal measurements, which increased temperatures, cocaine induced both hyperthermic and hypothermic responses. Direct comparisons revealed that body temperatures measured by a rectal probe are typically lower (approximately 0.6 degrees C) and more variable than body temperatures recorded by chronically implanted electrodes; the difference is smaller at low and greater at high basal temperatures. Because of this difference and temperature increases induced by the rectal probe per se, cocaine had no significant effect on rectal temperatures compared to control animals exposed to repeated rectal probes. Therefore, although rectal temperature measurements provide a decent correlation with directly measured deep body temperatures, the arousing influence of this procedure may drastically modulate the effects of other arousing stimuli and drugs. PMID- 17466280 TI - Purpurin expression in the zebrafish retina during early development and after optic nerve lesion in adults. AB - Purpurin, a retina-specific protein, is known to play a role in cell adhesion during development of the chicken retina. Although purpurin has been significantly detected in adult chicken retina, its function in the matured retina is not well understood. Therefore, to determine the expression pattern of purpurin in the retina, we simultaneously investigated expression patterns of purpurin in the zebrafish retina during development in larvae and optic nerve regeneration after nerve transection in adults. In early development, levels of purpurin suddenly increased in the zebrafish retina 3 to 5 days after fertilization, and purpurin-positive immunoreactivity was diffusely located in all retinal layers. In contrast, levels of purpurin mRNA rapidly increased in the adult retina 1-3 days after optic nerve transection, and rapidly declined by 10 days after injury. Signal for purpurin mRNA was seen only in photoreceptors. Immunohistochemistry showed that levels of purpurin protein were also increased in the retina 1-3 days after nerve injury, but positive staining was located in photoreceptors and ganglion cells, and the staining in ganglion cells was stronger than that in photoreceptors. Thus, the transient expression of purpurin protein was greatly different during development and optic nerve regeneration. In the former, purpurin may be required in all retinal layers, whereas in the latter, purpurin may be required for injured ganglion cells. PMID- 17466281 TI - What makes a discourse constraining? Comparing the effects of discourse message and scenario fit on the discourse-dependent N400 effect. AB - A discourse context provides a reader with a great deal of information that can provide constraints for further language processing, at several different levels. In this experiment we used event-related potentials (ERPs) to explore whether discourse-generated contextual constraints are based on the precise message of the discourse or, more 'loosely', on the scenario suggested by one or more content words in the text. Participants read constraining stories whose precise message rendered a particular word highly predictable ("The manager thought that the board of directors should assemble to discuss the issue. He planned a...[meeting]") as well as non-constraining control stories that were only biasing in virtue of the scenario suggested by some of the words ("The manager thought that the board of directors need not assemble to discuss the issue. He planned a..."). Coherent words that were inconsistent with the message-level expectation raised in a constraining discourse (e.g., "session" instead of "meeting") elicited a classic centroparietal N400 effect. However, when the same words were only inconsistent with the scenario loosely suggested by earlier words in the text, they elicited a different negativity around 400 ms, with a more anterior, left-lateralized maximum. The fact that the discourse-dependent N400 effect cannot be reduced to scenario-mediated priming reveals that it reflects the rapid use of precise message-level constraints in comprehension. At the same time, the left-lateralized negativity in non-constraining stories suggests that, at least in the absence of strong message-level constraints, scenario-mediated priming does also rapidly affect comprehension. PMID- 17466282 TI - Multimodal assessment of neuroprotection applied to the use of MK-801 in the endothelin-1 model of transient focal brain ischemia. AB - Transient focal ischemia produced by local infusion of endothelin-1 (ET1) in the territory of the middle cerebral artery has been proposed as a potentially useful model for the screening of drugs developed for the treatment of thrombo-embolic stroke. However, most of the data rely exclusively on the assessment of the infarct volume, which is only a partial predictor of the neurological outcome of stroke. Here, we have validated the model using a multimodal approach for the assessment of neuroprotection, which includes (i) determination of the infarct volume by 2,3,5-triphenyltetrazolium chloride staining; (ii) an in-depth behavioral analysis of the neurological deficit; and (iii) an EEG analysis of electrophysiological abnormalities in the peri-infarct somatosensory forelimb cortical area, S1FL. The non-competitive NMDA receptor antagonist, MK-801 (3 mg/kg, injected i.p. 20 min after ET1 infusion in conscious rats) could reduce the infarct volume, reverse the EEG changes occurring at early times post-ET1, and markedly improve the neurological deficit in ischemic animals. The latter effect, however, was visible at day 3 post-ET1, because the drug itself produced substantial behavioral abnormalities at earlier times. We conclude that a multimodal approach can be applied to the ET1 model of focal ischemia, and that MK-801 can be used as a reference compound to which the activity of safer neuroprotective drugs should be compared. PMID- 17466283 TI - The reduction of proprioceptors in the mesencephalic trigeminal tract nucleus after neonatal masseteric nerve transection; effect of brain-derived neurotrophic factor. AB - The effect of neonatal masseteric nerve transection on primary proprioceptors was examined in the mesencephalic trigeminal tract nucleus (Mes5) of the rat. At 72 h to 21 days after the injury, the number of Mes5 neurons decreased on the side ipsilateral to the transection. The means+/-SD of percentage proportion of ipsilateral/contralateral neurons at 72 h and 21 days were 69.9+/-7.5% and 58.2+/ 14.6%, respectively. The application of brain-derived neurotrophic factor to the proximal stump of the masseteric nerve delayed the loss of Mes5 neurons at 72 h after the injury; the mean numbers+/-SD of ipsilateral and contralateral Mes5 neurons in injured animals with BDNF application was 553.6+/-61.9 and 558.4+/ 55.3, respectively. Saline application had no effect on the injury-induced loss of Mes5 neurons; i.e., the mean numbers+/-SD of ipsilateral and contralateral Mes5 neurons were 367.3+/-72.5 and 543+/-33.5, respectively. These findings indicate that trigeminal primary proprioceptors are sensitive to the neonatal injury. The survival of proprioceptors during early postnatal period is probably dependent upon brain-derived neurotrophic factor in the trigeminal nervous system. PMID- 17466284 TI - Iris development in vertebrates; genetic and molecular considerations. AB - The iris plays a key role in visual function. It regulates the amount of light entering the eye and falling on the retina and also operates in focal adjustment of closer objects. The iris is involved in circulation of the aqueous humor and hence functions in regulation of intraocular pressure. Intriguingly, iris pigmented cells possess the ability to transdifferentiate into different ocular cell types of retinal pigmented epithelium, photoreceptors and lens cells. Thus, the iris is considered a potential source for cell-replacement therapies. During embryogenesis, the iris arises from both the optic cup and the periocular mesenchyme. Its interesting mode of development includes specification of the peripheral optic cup to a non-neuronal fate, migration of cells from the surrounding periocular mesenchyme and an atypical formation of smooth muscles from the neuroectoderm. This manner of development raises some interesting general topics concerning the early patterning of the neuroectoderm, the specification and differentiation of diverse cell types and the interactions between intrinsic and extrinsic factors in the process of organogenesis. In this review, we discuss iris anatomy and development, describe major pathologies of the iris and their molecular etiology and finally summarize the recent findings on genes and signaling pathways that are involved in iris development. PMID- 17466286 TI - Chronic fasting-induced changes of neuropeptide Y immunoreactivity in the lateral septum of intact and ovariectomized female rats. AB - The effect of 40% food deprivation for 1 week on the immunohistochemically detectable amount of neuropeptide Y (NPY) was studied in the lateral septum (LS) of intact and ovariectomized (OVX) female rats. Animals were either fed ad libitum or 40% food-deprived. Densitometric analysis of immunostained material showed a significant decrease in NPY-immunoreactivity (NPY-IR) in OVX rats compared to the control group. Food deprivation increased the density of punctate NPY-IR profiles in both intact and OVX animals, however, the density in food deprived OVX animals was increased compared to baseline but remained reduced compared to intact rats. Our study indicates that the lack of gonadal hormones - most likely estrogen - results in a decrease in the density of NPY-IR axonal fibers within the LS, while food deprivation induced considerable elevation in NPY density. Food restriction-induced changes in the density of NPY-containing neural elements suggest that the LS may play a crucial role in the regulation of food intake and energy balance, in concert with the relevant hypothalamic areas. PMID- 17466285 TI - Brain orexins and wake regulation in rats exposed to maternal deprivation. AB - Maternal deprivation (MD) is a neonatal stressor that leads to behavioral and molecular manifestations of chronic stress in adulthood. Recent evidence has suggested that stress may impact wake regulation through corticotropin-releasing hormone (CRH) and the orexinergic system. We studied the wake/sleep features and brain levels of orexin and orexin receptors in adult rats neonatally subjected to either ten days of MD or a control procedure from postnatal day 4. At 3 months of age, one set of rats from both groups underwent 48 h of polysomnographic recording. All rats (including those that did not undergo surgery) were subsequently sacrificed for ELISA, radioimmunoassay and western blot measurement of orexins, orexin receptors and CRH in multiple brain regions. Neonatal MD induced an increase of total wake time (decreased total sleep) during the light period, which corresponds to human night time. This increase was specifically composed of quiet wake, while a small but significant decrease of active wake was observed during the dark period. At the molecular level, MD led to increased hypothalamic CRH and orexin A, and frontal cortical orexin 1 receptors (OX1R). However, hippocampal orexin B was reduced in the MD group. Our study discovered for the first time that the adult MD rat has sleep and neurobiological features of hyperarousal, which is typical in human insomnia. We concluded that neonatal MD produces adult hyperarousal in sleep physiology and neurobiology, and that the adult MD rat could be a model of insomnia with an orexinergic mechanism. PMID- 17466287 TI - The solution conformation of C-glycosyl analogues of the sialyl-Tn antigen. AB - The conformational behavior of two C-glycosyl analogues of the sialyl-Tn antigen has been determined by a combination of NMR methods and molecular mechanics calculations. Both compounds show a major solution conformation that is drastically different from the major one of the natural compound. PMID- 17466288 TI - Impact of the extrusion process on xanthan gum behaviour. AB - Processing xanthan gum by extrusion and subsequent drying produces a biopolymer showing particulate, rather than molecular behaviour in aqueous solution. This form of xanthan disperses very readily to give a viscosity that is strongly dependent on salt concentration. On heating above the temperature of the order disorder transition as determined by calorimetry, there is a viscosity transition that is indicative of the irreversible loss of the particulate structure. It is suggested that the extrusion process melts and aligns xanthan macromolecules. On cooling reordering will occur but in the highly concentrated environment in the extruder ( approximately 45% water w/w), inter-molecular association between neighbouring macromolecules cannot proceed to completion due to kinetic trapping. As a consequence a network structure is created maintained by associations involving ordered regions. A xanthan solution can be prepared from this particulate material by dispersing and subsequent heating far more readily than can be achieved with non-processed xanthan. PMID- 17466289 TI - Synthesis of non-glycosidic 4,6'-thioether-linked disaccharides as hydrolytically stable glycomimetics. AB - Michael addition of 1,2:3,4-di-O-isopropylidene-6-thio-alpha-D-galactose (2) to 2 propyl 6-O-acetyl-3,4-dideoxy-alpha-D-glycero-hex-3-enopyranosid-2-ulose (1) afforded, as the major diastereoisomer, 2-propyl 6-O-acetyl-3-deoxy-4-S-(6-deoxy 1,2:3,4-di-O-isopropylidene-alpha-D-galactopyranos-6-yl)-4-thio-alpha-D-threo hexopyranosid-2-ulose (3, 91% yield). Reduction of the carbonyl group of 3, followed by O-deacetylation gave the two epimers 7 (alpha-D-lyxo) and 8 (alpha-D xylo) in a 1:2 ratio. On removal of the protecting groups of 8 by acid hydrolysis, formation of an 1,6-anhydro bridge was observed in the 3-deoxy-4 thiohexopyranose unit (10). The free non-glycosidic thioether-linked disaccharide 3-deoxy-4-S-(6-deoxy-alpha,beta-D-galactopyranos-6-yl)-4-thio-alpha,beta-D-xylo hexopyranose (11) was obtained by acetolysis of 10 followed by O-deacetylation. A similar sequence starting from the enone 1 and methyl 2,3,4-tri-O-benzoyl-6-thio alpha-D-glucopyranoside (12) led successfully to 2-propyl 3-deoxy-4-S-(methyl 6 deoxy-alpha-D-glucopyranos-6-yl)-4-thio-alpha-D-lyxo-hexopyranoside (17) and its alpha-D-xylo analog (19, major product). In this synthetic route, orthogonal sets of protecting groups were employed to preserve the configuration of both reducing ends and to avoid the formation of the 1,6-anhydro ring. PMID- 17466290 TI - Synthesis of substituted septanosyl-1,2,3-triazoles. AB - A carbohydrate-based oxepine, derived from 2-deoxy-D-arabino-hexopyranose, was used to prepare a family of septanosyl-1,2,3-triazoles in four steps. DMDO mediated epoxidation of the oxepine followed by trapping of the intermediate 1,2 anhydroseptanose by sodium azide gave the beta-substituted glycosyl azide. The septanosyl azide was then reacted with a number of alkynes under thermal Huisgen or copper(I) mediated reaction conditions. Hydrogenolysis of benzyl protecting groups gave substituted septanosyl-1,2,3-triazoles. The new septanose-based structures were then evaluated as potential glycosidase inhibitors. PMID- 17466291 TI - Calcium acts as a first messenger through the calcium-sensing receptor in the cardiovascular system. AB - It is well known that calcium is an important second messenger in the cardiovascular system. However, recent studies suggest that, in addition to its many functions as an intracellular messenger, Ca(2+) may also be an extracellular first messenger through the calcium-sensing receptor (CaR). The CaR belongs to family C of the G-protein-coupled receptors, which are also known as seven transmembrane domain receptors. The CaR receptor is expressed in all major organs involved in Ca(2+) homeostasis. Furthermore, increasing evidence suggests that the CaR is also involved in regulating various cellular functions in tissues not involved in Ca(2+) homeostasis. Recently, expression of a functional CaR has also been reported in crucial components of the cardiovascular system. It has previously been shown that the CaR is functionally expressed in the atria and ventricle of the rat heart. In blood vessels, the CaR protein was first reported in perivascular nerves of rat mesenteric resistance arteries, and was proposed to modulate myogenic tone in the arteries. Since then, the CaR has been detected in homogenates of whole vessels from rat subcutaneous small arteries and in endothelial cells from rat mesenteric and porcine coronary arteries. Furthermore, a recent report demonstrated that the CaR is present in endothelial cells from human aorta and that it stimulates production of nitric oxide in these cells. Taken together, these results indicate that the CaR present in blood vessels may have a physiological role in modulation of arterial blood pressure. This review discusses CaR expression and function, with a focus on the role of the CaR in the cardiovascular system. PMID- 17466292 TI - Oxidative stress expression status associated to Helicobacter pylori virulence in gastric diseases. AB - OBJECTIVES: To analyze the status of expression of inflammation markers, antioxidant and oxidant enzymes in biopsies from patients diagnosed with gastritis, gastric ulcer (GU) and gastric cancer (GC) and the Helicobacter pylori virulence from these isolated biopsies in order to evaluate a possible association among these factors. METHODS: H. pylori genotype from isolated biopsies was performed by PCR. The pattern of expression of inflammation (TNF alpha, IL-1beta, IL-8, IL-10 and IL-12), oxidant (iNOS and Nox1) and antioxidant markers (MnSOD, GPX and CAT) of biopsies from gastritis, GU, GC and control groups was performed by RT-PCR. RESULTS: Different from other gastric diseases studied here, gastritis is characterized by an oxidative stress with significant expression of TNF-alpha, IL-8, IL-12, iNOS and Nox and significant absence of MnSOD and GPX expression. Gastritis was the only condition where there was an association between TNF-alpha or IL-8 expression and H. pylori cagA+/vacAs1 genotype. In this case, TNF-alpha expression was about 3 times higher when compared to control subjects. CONCLUSION: In this study, only gastritis was found to be associated with significant oxidative stress marker expression of TNF-alpha and IL-8 that was also related to H. pylori virulence, suggesting that they are the main oxidant stress markers responsible to trigger an increase in ROS level that contributes to decrease the expression of the MnSOD and GPX. PMID- 17466293 TI - Inhibition of the alpha9alpha10 nicotinic cholinergic receptor by neramexane, an open channel blocker of N-methyl-D-aspartate receptors. AB - In this study we report the effects of neramexane, a novel amino-alkyl cyclohexane derivative that is a non-competitive N-methyl-D-aspartate (NMDA) receptor antagonist, on recombinant rat alpha9alpha10 nicotinic acetylcholine receptors expressed in Xenopus laevis oocytes. We compared its effects with those of memantine, a well-studied pore blocker of NMDA receptors, currently used in therapeutics for the treatment of Alzheimer's disease. Our results indicate that both compounds block acetylcholine-evoked responses at micromolar concentrations with a rank order of potency of neramexane>memantine, P<0.05. Block by neramexane of acetylcholine responses was not overcome at high concentrations of the agonist, indicative of a non-competitive inhibition. The lack of interaction of neramexane with the ligand binding domain was confirmed by radioligand binding experiments in transfected tsA201 cells. Moreover, block did not involve an increase in desensitization kinetics, it was independent of the resting potential of the membrane at low concentrations of neramexane and slightly voltage dependent at concentrations higher than 1 microM. Finally, clinically-relevant concentrations of neramexane blocked native alpha9alpha10-containing nicotinic acetylcholine receptors of rat inner hair cells, thus demonstrating a possible in vivo relevance in potentially unexplored therapeutic areas. PMID- 17466294 TI - Alpha1-adrenoceptors down-regulate ClC-2 chloride channels in epithelial cells from the acutely denervated jejunum. AB - Acute sympathetic denervation of the small intestine up-regulates alpha1 adrenoceptors on villus enterocytes and activation of these alpha1-adrenoceptors inhibits chloride secretion. We tested whether alpha1-adrenoceptor-mediated inhibition of chloride secretion was the result of reduced ClC-2 chloride channel expression. Phorbol myristate acetate (PMA) (a protein kinase C (PKC) activator) had no effect on ClC-2 levels. In contrast, alpha1-adrenoceptor activation significantly decreased ClC-2 protein levels in both the villus (1.58+/-0.19 to 0.75+/-0.19 arbitrary units) and crypt (1.69+/-0.15 to 0.37+/-0.23 arbitrary units) epithelial cells from the acutely denervated jejunum but not innervated controls. These data suggest that inhibition of chloride secretion following alpha1-adrenoceptor activation in the acutely denervated small intestine may be through ClC-2 down-regulation. PMID- 17466295 TI - Role for Btg1 and Btg2 in growth arrest of WEHI-231 cells through arginine methylation following membrane immunoglobulin engagement. AB - Engagement of membrane Ig (mIg) on WEHI-231 murine B lymphoma cells, a cell line model representative of primary immature B cells, results in growth arrest and subsequent apoptosis. Of the several dozen genes upregulated greater than two fold by anti-IgM treatment through DNA microarray analysis, we focused on B cell translocation gene 1 (Btg1) and Btg2, member of Btg/Tob family of proteins. WEHI 231 cells were infected with the Btg1/EGFP or Btg2/EGFP retroviral vectors, and those expressing either Btg1 or Btg2 accumulated in G1 phase at significantly higher proportions than that seen for cells expressing control vector. Btg1 or Btg2 bound to protein arginine methyltransferase (PRMT) 1 via the box C region, an interaction required for anti-IgM-induced growth inhibition. The arginine methyltransferase inhibitor AdOx partially abrogated growth inhibition induced by Btg1, Btg2, or anti-IgM. The Btg1- or Btg2-induced growth inhibition was also abrogated in PRMT1-deficient cells via introduction of small interference RNA. In addition, we observed anti-IgM-induced arginine methylation of two proteins, a 28 kDa and a 36-kDa protein. Methylation, detected by a monoclonal antibody specific for asymmetric, but not symmetric methyl residues, was observed as early as 1 h-2 h after stimulation and was sustained for up to 24 h. The anti-IgM-induced p36 arginine methylation was abrogated in the PRMT1-deficient cells, suggesting that PRMT1 induces p36 methylation. Together, these results suggest that anti-IgM induced growth inhibition is mediated via upregulation of Btg1 and Btg2, resulting in the activation of arginine methyltransferase activity and culminating in growth inhibition of WEHI-231 cells. PMID- 17466296 TI - Taenia solium and Taenia saginata: identification of sequence characterized amplified region (SCAR) markers. AB - Cysticercosis is one of the most important zoonosis, not only because of the effects on animal health and its economic consequences, but also due to the serious danger it poses to humans. The two main parasites involved in the taeniasis-cysticercosis complex in Brazil are Taenia saginata and Taenia solium. Differentiating between these two parasites is important both for disease control and for epidemiological studies. The purpose of this work was to identify genetic markers that could be used to differentiate these parasites. Out of 120 oligonucleotide decamers tested in random amplified polymorphic DNA (RAPD) assays, 107 were shown to discriminate between the two species of Taenia. Twenty one DNA fragments that were specific for each species of Taenia were chosen for DNA cloning and sequencing. Seven RAPD markers were converted into sequence characterized amplified region (SCAR) markers with two specific for T. saginata and five specific for T. solium as shown by agarose gel electrophoresis. These markers were developed as potential tools to differentiate T. solium from T. saginata in epidemiological studies. PMID- 17466297 TI - Pushing, pulling and trapping--modes of motor protein supported protein translocation. AB - Protein translocation across the cellular membranes is an ubiquitous and crucial activity of cells. This process is mediated by translocases that consist of a protein conducting channel and an associated motor protein. Motor proteins interact with protein substrates and utilize the free energy of ATP binding and hydrolysis for protein unfolding, translocation and unbinding. Since motor proteins are found either at the cis- or trans-side of the membrane, different mechanisms for translocation have been proposed. In the Power stroke model, cis acting motors are thought to push, while trans-motors pull on the substrate protein during translocation. In the Brownian ratchet model, translocation occurs by diffusion of the unfolded polypeptide through the translocation pore while directionality is achieved by trapping and refolding. Recent insights in the structure and function of the molecular motors suggest that different mechanisms can be employed simultaneously. PMID- 17466298 TI - Expression of adiponectin in choroidal tissue and inhibition of laser induced choroidal neovascularization by adiponectin. AB - The aim of this study was to investigate the role of adiponectin (APN) in a mouse model of laser induced choroidal neovascularization (CNV). We have shown by immunohistochemistry that the expression of APN, adiponectin receptor 1, adiponectin receptor 2 and T cadherin gradually increased from day 1 to day 7 post-laser in laser treated mice compared to controls. Recombinant APN (rAPN) was injected intraperitoneally (i.p., 25 microg/mouse) or intravitreally (2 microg/eye) in lasered mice. Another set of lasered mice received APN peptide via i.p. (75 microg/mouse) or intravitreal (30 microg/eye) route. Control mice received a similar treatment with PBS, control protein or control peptide after laser treatment. We found that in the i.p. and intravitreal injection of rAPN resulted in 78% and 68% inhibition respectively in the size of CNV complex compared to control mice. Similar results were observed when APN peptide was injected intravitreally or i.p. Treatment with rAPN or the peptide resulted in decreased levels of vascular endothelial growth factor. Thus, APN inhibited choroidal angiogenesis and may have therapeutic implications in the treatment of wet age related macular degeneration. PMID- 17466299 TI - FIGO Profile. Allan Rosenfield, M.D. PMID- 17466300 TI - Popularizing labor analgesia in China. AB - In China many women in labor are young primigravidas whose fear of labor pain leads them to request cesarean deliveries. While the rate of cesarean deliveries has reached 50% in many hospitals, less than 1% of women in labor are given neuraxial analgesia. The necessary equipment is seldom available in China and many physicians have misconceptions about the risks associated with neuraxial analgesia, which are low with the ultra-low-dosages used today. However, attitudes have begun to change. Meetings held in China have brought together Chinese physicians and world experts on the various epidural and combined spinal epidural techniques. Thanks to the information and support provided at these meetings clinical trials were carried out, more than 5000 women benefited from labor analgesia, and publications appeared in Chinese journals. An effective, safe, and cost-effective way to provide analgesia to women in labor may slow the increase in cesarean delivery rates across China and improve women's health in general. PMID- 17466301 TI - Routine second-trimester ultrasound for low risk pregnancies in a South African community. PMID- 17466302 TI - Perineometer and digital examination for assessment of pelvic floor strength. PMID- 17466303 TI - Reducing maternal mortality due to elective abortion: Potential impact of misoprostol in low-resource settings. AB - Over 99% of deaths due to abortion occur in developing countries. Maternal deaths due to abortion are preventable. Increasing the use of misoprostol for elective abortion could have a notable impact on maternal mortality due to abortion. As a test of this hypothesis, this study estimated the reduction in maternal deaths due to abortion in Africa, Asia and Latin America. The estimates were adjusted to changes in assumptions, yielding different possible scenarios of low and high estimates. This simple modeling exercise demonstrated that increased use of misoprostol, an option for pregnancy termination already available to many women in developing countries, could significantly reduce mortality due to abortion. Empirical testing of the hypothesis with data collected from developing countries could help to inform and improve the use of misoprostol in those settings. PMID- 17466304 TI - The Pap smear for detection of bacterial vaginosis. AB - OBJECTIVE: To assess validity of Pap smears in diagnosing bacterial vaginosis. METHOD: A prospective diagnostic accuracy study with 533 women in Mombasa, Kenya. Diagnosis of bacterial vaginosis using clinical observations scored with simplified Amsel's criteria and Bethesda system for Pap smears was compared with a reference standard (Nugent criteria for gram stains). Both laboratory tests were interpreted blindly. RESULT: Bacterial vaginosis prevalence was 36.7% (191/521) with Nugent criteria. Pap smear sensitivity and specificity were 59.4% (111/187) and 83.3% (270/324), with corresponding figures for simplified Amsel's criteria of 44.8% (81/181) and 84.8% (263/310). For Pap smear and simplified Amsel's criteria, positive predictive values were 67.3 and 63.3%, and negative predictive values 78.0% and 72.5%. CONCLUSION: In diagnosing bacterial vaginosis, Pap smears have moderate sensitivity (though higher than simplified Amsel's criteria). Specificity of Pap smears is adequate. Including bacterial vaginosis assessment as a standard component of Pap smears warrants consideration. PMID- 17466305 TI - Echolucent carotid plaques predict in-stent restenosis after bare metal stenting in native coronary arteries. AB - Echolucent carotid plaque is considered to predict coronary events. This study examined whether echolucent carotid plaque may predict in-stent restenosis (ISR) in coronary arteries. This study included 202 patients who had elective and successful percutaneous coronary intervention (PCI) with bare metal stents in de novo lesions of native coronary arteries for symptomatic coronary artery disease (CAD). Carotid plaque echolucency was assessed by ultrasound with integrated backscatter (IBS) analysis (intima-media IBS value minus adventitia IBS) 1 day before PCI. All patients underwent planned coronary angiography (CAG) at 6 months after PCI, or CAG before 6 months due to acute coronary syndromes. ISR (defined as >50% diameter stenosis) was found in 65 (32%) patients. The calibrated IBS values of carotid plaques were inversely correlated with late luminal loss of the stented lesions. Using multivariate logistic regression analysis, the presence of echolucent carotid plaques (