Impact of Chlorinated Water on Pathogen Inactivation during Wheat Tempering and Resulting Flour Quality.
Outbreaks of enteric pathogens linked to wheat flour have led the wheat milling industry to seek solutions addressing this food safety concern. Chlorinated water at 400 to 700 ppm has been used in the flour milling industry as a tempering aid to control growth of yeast and mold in tempering bins. However, the effectiveness of chlorinated water for inactivating enteric pathogens on wheat kernels was unknown. Five strains of Shiga toxin-producing Escherichia coli and two strains of Salmonella were inoculated onto hard red spring wheat at 7 log CFU/g and stored at room temperature for 1 month. Inoculated wheat was tempered with four concentrations (0, 400, 800, and 1,200 ppm) of chlorinated water (pH 6.5). The reduction due to chlorine was determined by calculating change in microbial loads at each chlorine level by using the response at 0 ppm as a reference. Uninoculated wheat tempered with chlorinated water was used to measure flour quality parameters. Changes in pathogen population over 18 h ranged from -2.35 to -0.30 log CFU/g with 800 ppm of chlorinated water and were not significantly different from changes at 400 and 1,200 ppm. Significant (P < 0.05) differences in the extent of reduction were observed among strains. However, the effect of chlorinated water at reducing native microbes on wheat kernels was minimal, with an average reduction of 0.39 log CFU/g for all concentrations. No significant (P > 0.05) changes occurred in flour quality and gluten functionality or during bread making for grains tempered at 400 and 800 ppm of chlorinated water. There were small but significant (P < 0.05) changes in flour protein content, final viscosity, and water absorption when tempered with 1,200 ppm of chlorinated water. The data showed that the level of chlorinated water currently used in industry for tempering could reduce enteric pathogen numbers by 1.22 log CFU/g for Shiga toxin-producing Escherichia coli and 2.29 log CFU/g for Salmonella, with no significant effects on flour quality and gluten functionality.


Views of community health workers on the integration of a physiotherapist into a ward-based outreach team.
Due to changes in the disease profile and lifestyle of individuals in South Africa, the limited health care facilities available have experienced congestion and overcrowding, affecting health care service delivery. Ward-based outreach team (WBOT) programmes were implemented to strengthen primary health care, improve access and alleviate the congestion occurring at these facilities. However, WBOTs have limitations in terms of medical knowledge and rehabilitative skills. To explore the views of community health workers (CHWs) on the integration of physiotherapists into WBOTs. A qualitative research design making use of focus group discussions (FGDs) was used. Through purposive sampling, 58 CHWs who were members of WBOTs were recruited. The WBOTs were from 10 selected primary health care centres in the Tshwane district, Region 2. Six FGDs were conducted. The audio-recorded data were transcribed verbatim. The transcripts were transported into NVivo 12 for thematic analysis. The views of the CHWs were that the WBOTs can benefit from having a physiotherapist as a member of the team. The WBOTs do not have adequate skills to attend to the physiotherapy needs of communities. People in the community have challenges in accessing physiotherapy services, and physiotherapy services can enhance the performance of the WBOTs by providing training to the WBOTs and providing clinical services to community members. Community-based rehabilitative services with a physiotherapist as part of the WBOTs can enhance and strengthen the services of the WBOTs, which can improve the treatment outcomes for communities. The WBOTS will be empowered to provide clinical services to the vulnerable people in the community that they serve.


Clomiphene citrate plus N-acetyl cysteine versus clomiphene citrate for augmenting ovulation in the management of unexplained infertility: a randomized double-blind controlled trial.
To compare clomiphene citrate with N-acetyl cysteine vs. clomiphene citrate alone for augmenting ovulation in management of unexplained infertility. Prospective randomized double-blind controlled trial. Department of obstetrics and gynecology in a university medical faculty in Egypt. Four hundred four patients as a study group (clomiphene citrate plus N-acetyl cysteine group) and 400 patients as a control group (clomiphene citrate-alone group). All women had unexplained infertility. Patients in the study group were treated with clomiphene citrate (50-mg tablets) twice per day and with N-acetyl cysteine (1,200 mg/d orally) for 5 days starting on day 2 of the cycle. Patients in the control group were treated with clomiphene citrate with sugar powder. The primary outcomes were number and size of growing follicles, serum E(2), serum P, and endometrial thickness. The secondary outcome was the occurrence of pregnancy. There were no statistically significant differences between the two groups in the number of follicles sized >18 mm, mean E(2) levels, serum P, or endometrial thickness. Pregnancy rate was comparable in both groups (22.2% vs. 27%). Miscarriage rate was comparable in both groups (6.7% in the study group vs. 7.4% in the control group). N-Acetyl cysteine is ineffective in inducing or augmenting ovulation in patients with unexplained infertility and cannot be recommended as an adjuvant to clomiphene citrate in such patients.


Estrogen receptor 1 and progesterone receptor are distinct biomarkers and prognostic factors in estrogen receptor-positive breast cancer: Evidence from a bioinformatic analysis.
Few prognostic indicators involving differentially expressed genes (DEGs) between estrogen receptor (ER)-positive and ER-negative breast cancer (BC) have been reported. We aimed to screen important DEGs related to ER status and explore potential prognostic factors for patients with ER-positive BC. Two microarray datasets (GSE22093 and GSE23988) downloaded from the Gene Expression Omnibus database were analyzed to identify DEGs between ER-positive and ER-negative BC tissue. Functional enrichment analysis of DEGs was performed using the Database for Annotation, Visualization, and Integrated Discovery server. Protein-protein interactions of the DEGs were analyzed using the Search Tool for the Retrieval of Interacting Genes. Subsequently, we studied the expression of hub genes in different histological types of BC using the Oncomine database. The online Kaplan-Meier (K-M) plotter survival analysis tool was utilized to evaluate the prognostic value of the expression of hub genes in ER-positive BC patients. Based on the results of K-M plotter analysis, we investigated the expression profiles of significant hub genes in an array of cancer cell lines using the Cancer Cell Line Encyclopedia database. A total of 194 DEGs were identified, comprising 141 upregulated and 53 downregulated genes. GO analysis revealed that the DEGs were mainly enriched in the extracellular exosome of the cellular component category. Ten hub genes were upregulated in ER-positive BC, and overexpression of estrogen receptor 1 (ESR1) mRNA was correlated with worse relapse-free survival (RFS). In contrast, overexpressed mRNA of progesterone receptor (PGR) was associated with longer RFS in patients with ER-positive BC. The expression of ESR1 was associated with GATA3, whereas that of PGR was correlated with ABLIM3. Both ESR1 and PGR were highly expressed in BC cell lines. The results suggest that mRNA expression levels of ESR1 and PGR can be considered distinct biomarkers and essential prognostic factors for ER-positive BC.


Detection and evaluation of myocardial fibrosis in Eisenmenger syndrome using cardiovascular magnetic resonance late gadolinium enhancement and T1 mapping.
Myocardial fibrosis is a common pathophysiological process involved in many cardiovascular diseases. However, limited prior studies suggested no association between focal myocardial fibrosis detected by cardiovascular magnetic resonance (CMR) late gadolinium enhancement (LGE) and disease severity in Eisenmenger syndrome (ES). This study aimed to explore potential associations between myocardial fibrosis evaluated by the CMR LGE and T1 mapping and risk stratification profiles including exercise tolerance, serum biomarkers, hemodynamics, and right ventricular (RV) function in these patients. Forty-five adults with ES and 30 healthy subjects were included. All subjects underwent a contrast-enhanced 3T CMR. Focal replacement fibrosis was visualized on LGE images. The locations of LGE were recorded. After excluding LGE in ventricular insertion point (VIP), ES patients were divided into myocardial LGE-positive (LGE+) and LGE-negative (LGE-) subgroups. Regions of interest in the septal myocardium were manually contoured in the T1 mapping images to determine the diffuse myocardial fibrosis. The relationships between myocardial fibrosis and 6-min walk test (6MWT), N-terminal pro-brain natriuretic peptide (NT-pro BNP), hematocrit, mean pulmonary arterial pressure (mPAP), pulmonary vascular resistance index (PVRI), RV/left ventricular end-systolic volume (RV/LV ESV), RV ejection fraction (RVEF), and risk stratification were analyzed. Myocardial LGE (excluding VIP) was common in ES (16/45, 35.6%), and often located in the septum (12/45, 26.7%). The clinical characteristics, hemodynamics, CMR morphology and function, and extracellular volume fraction (ECV) were similar in the LGE+ and LGE- groups (all P > 0.05). ECV was significantly higher in ES patients (28.6 ± 5.9% vs. 25.6 ± 2.2%, P < 0.05) and those with LGE- ES (28.3 ± 5.9% vs. 25.6 ± 2.2%, P < 0.05) than healthy controls. We found significant correlations between ECV and log NT-pro BNP, hematocrit, mPAP, PVRI, RV/LV ESV, and RVEF (all P < 0.05), and correlations trends between ECV and 6MWT (P = 0.06) in ES patients. An ECV threshold of 29.0% performed well in differentiating patients with high-risk ES from those with intermediate or low risk (area under curve 0.857, P < 0.001). Myocardial fibrosis is a common feature of ES. ECV may serve as an important imaging marker for ES disease severity.


CAD Severity on Cardiac CTA Identifies Patients With Most Benefit of Treating LDL-Cholesterol to ACC/AHA and ESC/EAS Targets.
This study aimed to assess if information on CAD severity from coronary computed tomography angiography (CTA) can identify patients that benefit most from treating low-density lipoprotein-cholesterol (LDL-C) to American Heart Association/American College of Cardiology (ACC/AHA) and European Society of Cardiology (ESC) guidelines targets. Current treatment guidelines for secondary prevention of atherosclerotic cardiovascular disease (ASCVD) disregard severity of coronary artery disease (CAD) for treatment choices. It is unclear whether severity of CAD should be considered in treatment recommendations. Among 20,241 symptomatic patients undergoing diagnostic CTA from the Western Denmark Heart Registry, we assessed the number needed to treat (NNT) in 6 years to prevent 1 ASCVD event as well as the proportion of all events that could be prevented by treating LDL-C to targets. We assumed a 22% relative reduction of ASCVD events per 1 mmol/l reduction in LDL-C. In multivariable analysis with no CAD as the reference, the subdistribution hazard ratio for ASCVD events was 4.0 (95% confidence interval [CI]: 3.3 to 4.9) for 1-vessel disease, 4.6 (3.5 to 6.0) for 2-vessel disease, and 5.6 (4.0 to 8.0) for 3-vessel disease. Consequently, the NNT to prevent 1 ASCVD event in 6 years by treating LDL-C to targets varied greatly from 233 (ESC) and 110 (ACC/AHA) for patients with no CAD to 8-9 for patients with 3-vessel disease (both ACC/AHA and ESC). The estimated percentage of ASCVD events that could be prevented by achieving guideline targets was 30% to 36% for patients with obstructive disease. However, <20% of patients achieved targets. An individualized approach based on CAD severity can identify symptomatic patients that are likely to derive most and least benefit from treating LDL-C to ACC/AHA and ESC treatment targets.


Chemical Biology Tools for Examining the Bacterial Cell Wall.
Bacteria surround themselves with cell walls to maintain cell rigidity and protect against environmental insults. Here we review chemical and biochemical techniques employed to study bacterial cell wall biogenesis. Recent advances including the ability to isolate critical intermediates, metabolic approaches for probe incorporation, and isotopic labeling techniques have provided critical insight into the biochemistry of cell walls. Fundamental manuscripts that have used these techniques to discover cell wall-interacting proteins, flippases, and cell wall stoichiometry are discussed in detail. The review highlights that these powerful methods and techniques have exciting potential to identify and characterize new targets for antibiotic development.


Macrophage-mediated immune response aggravates hearing disfunction caused by the disorder of mitochondrial dynamics in cochlear hair cells.
Mitochondrial dynamics is essential for maintaining the physiological function of the mitochondrial network, and its disorders lead to a variety of diseases. Our previous study identified mitochondrial dynamics controlled anti-tumor immune responses and anxiety symptoms. However, how mitochondrial dynamics affects auditory function in the inner ear remains unclear. Here, we show that the deficiency of FAM73a or FAM73b, two mitochondrial outer membrane proteins that mediate mitochondrial fusion, leads to outer hair cells (HCs) damage and progressive hearing loss in FVB/N mice. Abnormal mitochondrial fusion causes elevated oxidative stress and apoptosis of HCs in the early stage. Thereafter, the activation of macrophages and CD4+ T cell is found in the mutant mice with the increased expression of the inflammatory cytokines IL-12 and IFN-γ compared with control mice. Strikingly, a dramatically decreased number of macrophages by Clophosome®-A-Clodronate Liposomes treatment alleviates the hearing loss of mutant mice. Collectively, our finding highlights that FAM73a or FAM73b deficiency affects HCs survival by disturbing the mitochondrial function, and the subsequent immune response in the cochleae worsens the damage of HCs.


Developing an algorithm for pulse oximetry derived respiratory rate (RR(oxi)): a healthy volunteer study.
The presence of respiratory information within the pulse oximeter signal (PPG) is a well-documented phenomenon. However, extracting this information for the purpose of continuously monitoring respiratory rate requires: (1) the recognition of the multi-faceted manifestations of respiratory modulation components within the PPG and the complex interactions among them; (2) the implementation of appropriate advanced signal processing techniques to take full advantage of this information; and (3) the post-processing infrastructure to deliver a clinically useful reported respiratory rate to the end user. A holistic algorithmic approach to the problem is therefore required. We have developed the RR(OXI) algorithm based on this principle and its performance on healthy subject trial data is described herein. Finger PPGs were collected from a cohort of 139 healthy adult volunteers monitored during free breathing over an 8-min period. These were subsequently processed using a novel in-house algorithm based on continuous wavelet transform technology within an infrastructure incorporating weighted averaging and logical decision making processes. The computed oximeter respiratory rates (RR(oxi)) were then compared to an end-tidal CO2 reference rate RR(ETCO2). RR(ETCO2) ranged from a lowest recorded value of 2.97 breaths per min (br/min) to a highest value of 28.02 br/min. The mean rate was 14.49 br/min with standard deviation of 4.36 br/min. Excellent agreement was found between RR(oxi) and RR(ETCO2), with a mean difference of -0.23 br/min and standard deviation of 1.14 br/min. The two measures are tightly spread around the line of agreement with a strong correlation observable between them (R2 = 0.93). These data indicate that RR(oxi) represents a viable technology for the measurement of respiratory rate of healthy individuals.


Gender moderates the effect of exercise on anxiety sensitivity.
A moderate to vigorous intensity exercise program is emerging as a promising strategy for reducing anxiety sensitivity (AS). Initial evidence suggests that the effects of exercise on mental health outcomes may vary as a function of gender, with men benefitting more than women. Building upon this evidence, the present study tested the hypothesis that the effect of exercise on AS would vary as a function of gender, such that the effect would be stronger for men than for women. We tested this hypothesis using the data from a published study (Smits, Berry, Rosenfield, et al., 2008). In this study, participants (N = 60) with elevated levels of AS were randomly assigned to a two-week exercise intervention [EX] or a waitlist control condition [WL]. Results revealed that males showed significantly greater initial AS reductions relative to females (following 1 week of exercise). However, these gender differences were no longer evident at the end of the intervention. Possible mechanisms for the observed findings and directions for future research are discussed.


Secure mobile agent for telemedicine based on P2P networks.
Exploring intelligent mobile agent (MA) technology for assisting medical services or transmitting personal patient-health information in telemedicine applications has been widely investigated. Conversely, peer-to-peer (P2P) networking has become one of the most popular applications used in the Internet because of its benefits for easy-to-manage resources and because it balances workloads. Therefore, constructing an agent-based telemedicine platform based on P2P networking architecture is necessary. The main purpose of this paper is to construct a safe agent-based telemedicine that based on P2P networking architecture. Two themes are addressed in this paper: (a) the P2P network architecture for an agent-based telemedicine service, and (b) the security mechanisms for the proposed telemedicine networking architecture. When an MA contains patient information and migrates from one host to another through the Internet, it can be attacked by other software agents or agent platforms that can illegally access patient information. The proposed P2P network architecture is based on the JXTA protocol and provides two types of telemedicine service models: the predictable service model and unpredictable service model. This architecture employs a two-layer safety mechanism for MAs (i.e., time-limited black boxes and RSA undetachable signature technologies), to provide a secure solution for agent-based telemedicine services.


Author Correction: Anti-trypanosomatid drug discovery: an ongoing challenge and a continuing need.
The structures of nifurtimox in Table 1 were incorrect and have been updated in the pdf and online. The authors apologize for any confusion caused.


Immune recognition of syngeneic, allogeneic and xenogeneic stromal cell transplants in healthy retinas.
Advanced therapies using adult mesenchymal stromal cells (MSCs) for neurodegenerative diseases are not effectively translated into the clinic. The cross talk between the transplanted cells and the host tissue is something that, despite its importance, is not being systematically investigated. We have compared the response of the mouse healthy retina to the intravitreal transplantation of MSCs derived from the bone marrow in four modalities: syngeneic, allogeneic, xenogeneic and allogeneic with immunosuppression using functional analysis in vivo and histology, cytometry and protein measurement post-mortem. Data were considered significant (p < 0.05) after nonparametric suitable statistical tests. Transplanted cells remain in the vitreous and are cleared by microglial cells a process that is quicker in allotransplants regardless of immunosuppression. All transplants cause anatomical remodelling which is more severe after xenotransplants. Xeno- and allotransplants with or without immunosuppression cause macro- and microglial activation and retinal functional impairment, being xenotransplants the most detrimental and the only ones that recruit CD45+Iba1-cells. The profile of proinflammatory cytokines changes in all transplantation settings. However, none of these changes affect the retinal ganglion cell population. We show here a specific functional and anatomical retinal response depending on the MSC transplantation modality, an aspect that should be taken into consideration when conducting preclinical studies if we intend a more realistic translation into clinical practice.


The Association Between School Tobacco Control Policies and Youth Smoking Behavior.
While U.S. tobacco control policy has focused mainly on tobacco excise taxes, product advertising bans, and state tobacco control policies such as indoor/outdoor smoking bans, little attention has been paid to school tobacco control policies and their impact on youth smoking behavior. Thus, the objective of this study is to examine the impact of school tobacco control policies on smoking behavior among teenagers and young adults in the USA. Using logistic regression approach, this study examines the effect of school tobacco control policies on individuals ever trying smoking and ever being a regular smoker using data from waves I, II, and III of the National Longitudinal Study of Adolescent Health. Findings indicate that school tobacco control policies targeting both students and employees of the school are associated with a reduced odds of smoking initiation among youth, but do not have much effect on youth becoming regular smokers. If implemented properly, school tobacco control policies could play a vital role in preventing youth smoking and help reduce youth smoking rates in the country, addressing a key public health issue.


Rapid antigen detection in the diagnosis of highly pathogenic avian influenza (H5N1) virus in Nigeria.
Highly pathogenic avian influenza (AI) is an infectious disease of agroeconomic and public health importance. The outbreak that occurred in Nigeria (2006-2008) was devastating to the poultry industry and raised public health concerns. In the course of its control, rapid laboratory confirmation of suspected cases in poultry was essential for prompt mobilization of control logistics for depopulation and decontamination of affected premises. Commercial rapid test kit was evaluated in the diagnosis of highly pathogenic AI (HPAI) as a preliminary to virus isolation. Between 2006 and 2007, 382 cases were tested out of which 149 were positive by rapid antigen detection. Virus isolation yielded 171 positive cases. The relative diagnostic sensitivity of Anigen Rapid AIV Ag test was 84.3% (95% confidence interval [CI], 78.1-88.9%), whereas the relative diagnostic specificity was 97.7% (95% CI, 94.2-99.1%). Rapid antigen detection is a useful technique for prompt diagnosis of HPAI for early detection and containment.


Current concepts of the effect of oxygen on the developing retina.
The role of oxygen in the development of retrolental fibroplasia and the mechanism of oxygen changes in the developing retina are discussed. The primary effect of oxygen is one of Vascular closure and the secondary response, which occurs after removal from increased oxygen, is retinal neovascularization. A brief discussion of recent observations on the potential role of vitamin E in the prevention of retrolental fibroplasia is presented.


Angiomyolipoma of the liver: analysis of typical features and pitfalls based on own experience and literature.
We present imaging findings (ultrasound, computed tomography, and magnetic resonance imaging) of eight patients with hepatic angiomyolipoma (HAML). The lesions were solitary in seven patients, and one patient had multiple tumors (n=11). Angiomyolipoma, even though a rare liver tumor, should be included in the differential diagnosis in cases of highly vascularized lesion containing a significant amount of fat. Suggestion of the diagnosis of HAML might be helpful for the pathologist in the selection of the typical histochemical staining of the tumor, allowing accurate diagnosis, which, in turn, determines the implementation of appropriate therapeutic intervention.


Head and neck solitary fibrous tumors: diagnostic and therapeutic challenges.
Head and neck solitary fibrous tumors (SFTs) are quite rare, with <200 cases reported in the literature. The purpose of this study was to analyze the diagnostic and therapeutic challenges of this rare disease while reporting the largest head and neck case series. Retrospective review. Between 1991 and 2011, 13 cases of head and neck SFTs from the pathology database were identified. Demographic, clinical, radiologic, and pathologic data were then abstracted and compiled for review. SFT presented nearly equally as asymptomatic slow-growing masses or with local symptoms due to compression. All tumors imaged by computed tomography or magnetic resonance imaging with contrast showed a well-defined strongly enhancing mass. Two patients had evidence of bone erosion on imaging, but no histopathologic features of malignancy. Treatment consisted of surgical resection and in two cases postoperative radiation. Two patients had pathologic findings of malignancy, but are alive without recurrence at 26 and 51 months follow-up. Two patients had recurrence, one with paraspinal disease and grossly positive surgical margins, and another with endoscopic resection and poor margin control. SFTs of the head and neck are exceedingly rare and those with aggressive behavior even more so. Diagnosis is often difficult and not definitive until morphologic and immunohistochemical evaluation is performed. In most cases, complete surgical excision is the only necessary treatment. Although in patients with malignant components or positive surgical margins, adjuvant radiation may be beneficial. Regardless, all patients require close clinical follow-up for several years.


The contribution of cerebral vascular semicarbazide-sensitive amine oxidase to cerebral amyloid angiopathy in Alzheimer's disease.
Semicarbazide-sensitive amine oxidase (SSAO) catalyses the oxidative deamination of a variety of endogenous substrates, such as methylamine and aminoacetone, to produce highly reactive aldehydes, which are capable of inducing protein cross-linkage, beta amyloid (Abeta) aggregation and advanced glycation end-product formation. In the brain, SSAO is exclusively located on the blood vessels. Deposits of Abeta, the hallmark of Alzheimer's disease (AD), are closely associated with cerebral blood vessels, that is, cerebral amyloid angiopathy (CAA). In the present study, we examined whether SSAO-mediated deamination contributes to CAA in AD. We employed immunohistochemistry to examine the colocalization of SSAO and Abeta in post mortem brains of AD patients. To assess the role of SSAO-mediated deamination in the deposition of Abeta on blood vessel walls, we developed an in vitro blood vessel model using sections of human umbilical cord. We found a strong expression of SSAO colocalized with Abeta deposits on the blood vessels in AD brains. We also demonstrated that SSAO-mediated deamination increases the deposition of Abeta onto blood vessel walls. Our results support the hypothesis that cerebral vascular SSAO-catalysed deamination contributes to CAA in AD brains.


Effect of antibiotics on cellular stress generated in Shiga toxin-producing Escherichia coli O157:H7 and non-O157 biofilms.
Shiga toxin-producing Escherichia coli (STEC) are important food-borne pathogens, with the main virulence factor of this bacterium being its capacity to secrete Shiga toxins (Stxs). Therefore, the use of certain antibiotics for the treatment of this infection, which induces the liberation of Stxs, is controversial. Reactive oxygen and nitrogen species are also involved in the pathogenesis of different diseases. The purpose of this study was to analyze the effects of antibiotics on biofilms of STEC and the relationships between cellular stress and the release of Stx. To this end, biofilms of reference and clinical strains were treated with antibiotics (ciprofloxacin, fosfomycin and rifaximin) and the production of oxidants, the antioxidant defense system and toxin release were evaluated. Ciprofloxacin altered the prooxidant-antioxidant balance, with a decrease of oxidant metabolites and an increase of superoxide dismutase and catalase activity, being associated with high-levels of Stx production. Furthermore, inhibition of oxidative stress by exogenous antioxidants was correlated with a reduction in the liberation of Stx, indicating the participation of this phenomenon in the release of this toxin. In contrast, fosfomycin and rifaximin produced less alteration with a minimal production of Stx. Our data show that treatment of biofilm-STEC with these antibiotics induces oxidative stress-mediated release of Stx.


Post-operative myocardial dysfunction does not affect the physiological response to early mobilization after coronary artery bypass grafting.
An acute increase in oxygen demand can be compensated for either by increased cardiac index (CI) or increased oxygen extraction, resulting in reduced mixed venous oxygen saturation (SvO2). We tested the hypothesis that post-operative cardiac dysfunction may explain why oxygen extraction alone is increased during early mobilization after cardiac surgery. Twenty patients with a pre-operative ejection fraction > 50% were included in an open prospective observational study comparing the changes in SvO2 and hemodynamics during mobilizations immediately prior to surgery and on the first post-operative morning. Mobilization induced an absolute reduction in SvO2 of 17.7 +/- 7.4% pre- and 19.0 +/- 5.5% post-operatively (NS). ANOVA for a series of measurements throughout the mobilization sequence identified no different effect on SvO2 between pre- and post-operative mobilizations (P = 0.567). The SvO2 level was reduced post-operatively resulting in a SvO2 during standing exercise of 55% before and 49% after the surgery (P < 0.01). Mobilization increased the heart rate (HR) and decreased the stroke volume index (SVI), leaving CI unchanged. This response was similar pre- and post-operatively (NS). Compared with pre-operative measurements, CI and HR increased post-operatively while SVI remained unchanged despite elevated cardiac filling pressures and reduced systemic vascular resistance. The left ventricular stroke work index was reduced, indicating reduced myocardial performance. Myocardial function was reduced on the first morning after coronary artery bypass grafting (CABG), but during post-operative mobilization this reduction did not significantly influence the changes in CI or SvO2.


Finger-loop inhibitors of the HCV NS5b polymerase. Part 1: Discovery and optimization of novel 1,6- and 2,6-macrocyclic indole series.
Novel conformationaly constrained 1,6- and 2,6-macrocyclic HCV NS5b polymerase inhibitors, in which either the nitrogen or the phenyl ring in the C2 position of the central indole core is tethered to an acylsulfamide acid bioisostere, have been designed and tested for their anti-HCV potency. This transformational route toward non-zwitterionic finger loop-directed inhibitors led to the discovery of derivatives with improved cell potency and pharmacokinetic profile.


Tips of the trade #15. Removal of hardware prior to total hip replacement.
Since the introduction of metals into orthopaedic surgery over a hundred years ago, patients often present with a wide variety of hardware in their bones. Normally this is of little significance, but if the hardware is in the proximal femur and the hip is arthritic, then the metal must be removed before hip replacement can commence. This paper presents a series of guidelines to be used in such cases.


BDE-47 sorption and desorption to soil matrix in single- and binary-solute systems.
Three loamy-clay soil samples (LC1-3) with different properties were collected as the geosorbents to preliminarily investigate the sorption and desorption of 2,2',4,4'-tetrabromodiphenyl ether (BDE-47) in single system and binary system with the presence of decabromodiphenyl ether (BDE-209), which can provide information in order to further understand the sorption mechanisms and evaluate the adsorption sites. A concentration of 10 μg L(-1) BDE-209 suppressed the sorption of BDE-47, and the trend became more and more significant with the increase of BDE-47 equilibrium concentration, however, BDE-47 caused no competitive effect on BDE-209 sorption, which was related with the better accessibility of more hydrophobic molecules to adsorption sites. In the binary system, nonlinearity of the BDE-47 sorption isotherms for the three samples changed in different ways, which originated from the varied soil properties. Desorption hysteresis was observed in all cases, which was estimated due to irreversible surface adsorption between sorbent and sorbate. BDE-209 made desorption of BDE-47 more hysteretic from soil samples, which was estimated to be ascribed to the accelerated sorbent state transition and new sites creation caused by BDE-209 sorption.


Species cross-reactive Ia determinants: detection with a monoclonal antibody to human Ia of a murine Ia. 7 epitope.
A.TH anti-A.TL alloserum has previously been shown to react with monomorphic determinants of human Ia molecules. In an attempt to produce monoclonal antibodies detecting human-mouse cross-reacting epitopes, A.TH mice (Is) which lack I-E antigens were immunized with human Ia glycoproteins. Five hybridoma lines were established which recognize monomorphic human Ia determinants. Only one of the lines, 21w4, is reactive with murine cells and also with pig and sheep cells but not with rat cells. Binding studies to murine target cells show a strain distribution analogous to that of an Ia. 7-like specificity. The binding of 21w4 antibody to human and murine cells was compared to that of two other monoclonal antibodies directed to the murine Ia. 7-like specificity. Although all three antibodies had a similar pattern of reactivity with cells of different mouse strains, only 21w4 bound to the human cells tested. Competitive inhibition studies on murine cells have revealed that the epitopes were spatially related but not identical. Our results support the view that Ia. 7, the putative species cross-reacting specificity, might be composed of clusters of epitopes with variable degrees of conservation throughout evolution.


Population Pharmacokinetics and Bayesian Estimation of the Area Under the Concentration-Time Curve for Ganciclovir in Adult Chinese Renal Allograft Recipients After Valganciclovir Administration.
Valganciclovir (VGCV) is the prodrug of ganciclovir (GCV). The objective of this study was to establish a population pharmacokinetic model (PPK) of GCV to investigate the PK characteristics of GCV after administration of VGCV in adult Chinese renal allograft recipients. Seventy Chinese renal allograft recipients were given 450 mg (n = 41) or 900 mg (n = 29) VGCV daily. Blood samples were drawn 0-24 hours after 5 days' therapy, and GCV plasma levels were determined. The PPK model was constructed using nonlinear mixed-effects modeling, and the Bayesian estimation of AUC0-24h was constructed for an individual patient based on limited plasma samples. The PK of GCV was best described by a 2-compartment model with a first-order absorption process. The CL/F, V2 /F, Q/F, V3 /F, Ka , and lag time of GCV were 15.8 ± 0.71 L/h, 10.9 ± 2.38 L, 3.98 ± 0.40 L/h, 167 ± 44.0 L, 0.23 ± 0.0078 1/h, and 0.93 ± 0.017 hours, respectively. Clearance of creatinine was found to have a significant impact on the CL/F of GCV (P < .01). Sampling strategies consisted of plasma concentrations 0 and 2 and 0, 2, and 4 hours after VGCV administration were shown to be suitable for the estimation of the GCV AUC0-24h . The PPK model was acceptable and can describe the PK of GCV in Chinese renal transplant patients administered VGCV. The AUC0-24h of GCV in Chinese renal transplant patients can be calculated by a limited sampling strategy method.


Impact on human health of Salmonella spp. on pork in The Netherlands and the anticipated effects of some currently proposed control strategies.
The impact on human health of Salmonella spp. on pork in The Netherlands is described. Subsequently, the effects of some currently proposed control strategies in the Dutch pork production chain are evaluated and quantified with the aid of a simple mathematical model. The estimated average incidence of cases of salmonellosis in the Netherlands is about 450 cases per 100,000 person years at risk (pyar). Some special risk groups for which the risks could be quantified are (1) persons with underlying diseases, such as neoplasms or diabetes mellitus (1200 cases/100,000 pyar); (2) persons with achlorhydria or who excessively use antacids (1100 cases/100,000 pyar); (3) persons who have recently been treated with antibiotics that disturb the normal gut flora (1700 cases/100,000 pyar); (4) nurses (900 cases/100,000 pyar); (5) caterers (900 cases/100,000 pyar); (6) slaughterline personnel (1800 cases/100,000 pyar). Furthermore, it is estimated that 15% (5-25%) of all cases of salmonellosis in The Netherlands are associated with the consumption of pork. Currently, proposed control measures regarding Salmonella in pigs and on pork in The Netherlands are codes of good manufacturing practices (GMP) that, in fact, formalize recommendations that can be found in many handbooks about pig breeding and pig slaughtering. When evaluated by a mathematical model constructed for this purpose, the proposed GMP codes from farm to cutting/retail could, at best, reduce the current levels of Salmonella-positive pigs and pork by 50-60%. If pigs were bred according to the rather costly specific pathogen-free concept (SPF), the prevalence of contaminated carcasses and pork could in total be reduced by 95% or more. However, implementing GMP codes from the transport phase up to the cutting/retail phase coupled with a decontamination step at the end of the slaughterline would be just as effective as GMP in combination with breeding using the SPF-concept. It is therefore concluded that the most efficient and cost-effective way of reducing the 'Salmonella problem' entailed by the consumption of pork would be to decontaminate carcasses under the precondition that the entire production chain strictly adheres to GMP principles. Therefore, the EU should also allow for more possibilities regarding the decontamination of carcasses than is currently the case. It is also concluded that current EU regulations relying on hazard analysis of critical control points (HACCP)-inspired production in cutting plants will not be effective in reducing the prevalence of Salmonella spp. on pork. This is mainly because (1) there is currently an almost steady stream of Salmonella-positive carcasses that enter the cutting process; (2) when contaminated carcasses are being processed, further cross-contamination during working hours is unavoidable; (3) no steps in the cutting process are intentionally designed to effectively reduce the risks or consequences of cross contamination of cuts and retail-ready products.


Dimethyl fumarate: A review of preclinical efficacy in models of neurodegenerative diseases.
Dimethyl fumarate (DMF) is an antioxidative and anti-inflammatory drug approved for treatment of multiple sclerosis and psoriasis; however, beneficial effects of DMF have also been found in other inflammatory diseases and cancers. DMF is a prodrug that is immediately hydrolysed to monomethyl fumarate (MMF) in vivo. Both fumarates activate the nuclear factor (erythroid-derived 2)-like 2 (Nrf2) pathway, and Nrf2 is a key transcription factor of the antioxidant response. The immunosuppressive and anti-inflammatory actions of DMF occur through several mechanisms: via inhibition of the nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) pathway and by downregulation of aerobic glycolysis and pyroptosis in activated myeloid and lymphoid cells. MMF is also an agonist of hydroxycarboxylic acid receptor 2 (HCAR2). Differences in the strength of effects and mechanisms of action of both fumarates are discussed. The aim of this review was to analyse and compare the neuroprotective, antioxidative and anti-inflammatory effects of DMF and its active metabolite, MMF, in cellular (in vitro) and animal models of neurodegenerative diseases (NDs), other than multiple sclerosis. There are more than twenty studies that currently represent this field. Most of the studies are concerned with cellular or animal models of Alzheimer's disease (AD) and Parkinson's disease (PD), one utilized a mouse model of Huntington's disease (HD) and one clinical trial was carried out with amyotrophic lateral sclerosis (ALS) patients. The discrepancies in the results of the various studies are discussed, and issues requiring further research have been identified.


Parallel and interactive learning processes within the basal ganglia: relevance for the understanding of addiction.
In this review we discuss the evidence that drug addiction, defined as a maladaptive compulsive habit, results from the progressive subversion by addictive drugs of striatum-dependent operant and Pavlovian learning mechanisms that are usually involved in the control over behaviour by stimuli associated with natural reinforcement. Although mainly organized through segregated parallel cortico-striato-pallido-thalamo-cortical loops involved in motor or emotional functions, the basal ganglia, and especially the striatum, are key mediators of the modulation of behavioural responses, under the control of both action-outcome and stimulus-response mechanisms, by incentive motivational processes and Pavlovian associations. Here we suggest that protracted exposure to addictive drugs recruits serial and dopamine-dependent, striato-nigro-striatal ascending spirals from the nucleus accumbens to more dorsal regions of the striatum that underlie a shift from action-outcome to stimulus-response mechanisms in the control over drug seeking. When this progressive ventral to dorsal striatum shift is combined with drug-associated Pavlovian influences from limbic structures such as the amygdala and the orbitofrontal cortex, drug seeking behaviour becomes established as an incentive habit. This instantiation of implicit sub-cortical processing of drug-associated stimuli and instrumental responding might be a key mechanism underlying the development of compulsive drug seeking and the high vulnerability to relapse which are hallmarks of drug addiction.


Design and synthesis of some barbituric and 1,3-dimethylbarbituric acid derivatives: A non-classical scaffold for potential PARP1 inhibitors.
Six series based on barbituric acid 5a-e, 10a-d; thiobarbituric acid 6a-e, 11a-d and 1,3-dimethylbarbituric acid 7a-e, 12a-d were prepared and screened for their in vitro PARP1 inhibition. They revealed promising inhibition at nanomolar level especially compounds 5c, 7b, 7d and 7e (IC50 = 30.51, 41.60, 41.53 and 36.33 nM) with higher potency than olaparib (IC50 = 43.59 nM). Moreover, compounds 5b, 5d, 7a, 12a and 12c exhibited good comparable activity (IC50 = 65.93, 58.90, 66.57, 45.40 and 50.62 nM, respectively). Furthermore, the most active compounds 5c, 7b, 7d, 7e, 12a and 12c against PARP1 in vitro were evaluated in the BRCA1 mutated triple negative breast cancer cell line MDA-MB-436 where 5c and 12c showed higher potency compared to olaparib and result in cell cycle arrest at G2/M phase. 5c and 12c showed apoptotic effects in MDA-MB-436 and potentiated the cytotoxicity of temozolomide in A549 human lung epithelial cancer cell line. Compounds 5c and 12c represent interesting starting points towards PARP1 inhibitors.


Hemothorax due to vena caval erosion by a subclavian dual-lumen dialysis catheter.
We have reported a case of vena caval perforation by a polyurethane dual-lumen subclavian vein catheter unrelated to insertion or subsequent manipulation. This case emphasizes the necessity for constant vigilance while such catheters are in use.


Comparison of MiraLAX and magnesium citrate for bowel preparation at CT colonography.
To compare MiraLAX, a hypo-osmotic lavage, and magnesium citrate (MgC), a hyper-osmotic agent for bowel preparation at CTC. 398 total screening CTC studies were included in this retrospective, single institution study. 297 underwent preparation with a double-dose MgC regimen (mean age, 61 ± 5.5 years; 142 male/155 female) and 101 with 8.3 oz (equivalent to 238 g PEG) of MiraLAX (mean age, 60 ± 9.6 years; 45 male/56 female). Oral contrast for tagging purposes was utilized in both regimens. Studies were retrospectively analyzed for residual fluid volume and attenuation by automated analysis, as well for subjective oral contrast coating of the normal colonic wall and polyps. 50 patients underwent successive CTC studies utilizing each agent (mean, 6.1 ± 1.7 years apart), allowing for intra-patient comparison. Chi-squared, Fisher's exact, McNemar, and t-tests were used for data comparison. Residual fluid volume (as percentage of total colonic volume) and fluid density was 7.2 ± 4.2% and 713 ± 183 HU for the MgC cohort and 8.7 ± 3.8% and 1044 HU ± 274 for the MiraLAX cohort, respectively (p = 0.001 and p < 0.001, respectively). Similar results were observed for the intra-patient cohort. Colonic wall coating negatively influencing interpretation was noted in 1.7% of MgC vs. 6.9% of MiraLAX examinations (p = 0.008). Polyps were detected in 12% of all MgC vs. 16% of all MiraLAX CTCs (p = 0.29). CTC bowel preparation with the hypo-osmotic MiraLAX agent appears to provide acceptable diagnostic quality that is comparable to the hyper-osmotic MgC agent, especially when factoring in patient safety and tolerance.


Notch2 genetic fate mapping reveals two previously unrecognized mammary epithelial lineages.
Notch signalling is implicated in stem and progenitor cell fate control in numerous organs. Using conditional in vivo genetic labelling we traced the fate of cells expressing the Notch2 receptor paralogue and uncovered the existence of two previously unrecognized mammary epithelial cell lineages that we term S (Small) and L (Large). S cells appear in a bead-on-a-string formation and are embedded between the luminal and basal/myoepithelial layers in a unique reiterative pattern, whereas single or paired L cells appear among ductal and alveolar cells. Long-term lineage tracing and functional studies indicate that S and L cells regulate ipsi- and contralateral spatial placement of tertiary branches and formation of alveolar clusters. Our findings revise present models of mammary epithelial cell hierarchy, reveal a hitherto undescribed mechanism regulating branching morphogenesis and may have important implications for identification of the cell-of-origin of distinct breast cancer subtypes.


Effects of the novel dihydropyridine BAY-K-8644 on adrenomedullary catecholamine release evoked by calcium reintroduction.
Reintroduction of Ca ions to cat adrenal glands perfused at room temperature with Krebs solution lacking Ca and Mg, evoked a catecholamine secretory response that was directly proportional to the concentration of Ca reintroduced. This secretory response inactivated quickly, was abolished by nM concentrations of nifedipine and was potentiated dramatically by nM concentrations of BAY-K-8644. Excess Ca antagonized the inhibitory effects of nifedipine and this drug inhibited competitively the potentiating effects of BAY-K-8644. These data suggest 1st) that extracellular divalent cations deprivation activates specific Ca channels; 2nd) that the dihydropyridine BAY-K-8644 increases the release of catecholamines by Ca reintroduction by activating and/or delaying the inactivation of Ca channels; and 3rd) that the access of the dihydropyridine-type Ca agonist and antagonists to their "intra-channel" site of action requires the pre-activation of Ca channels.


Differentiation among populations of Sedum wrightii (Crassulaceae) in response to limited water availability: water relations, CO2 assimilation, growth and survivorship.
Sedum wrightii is one of only a few species in the Crassulaceae for which there is evidence for a high degree of variability in the ratio of daytime to nighttime CO2 assimilation. There are both environmental and genetic components to this variability. S. wrightii grows over a wide altitudinal gradient. The purpose of this study was to compare low, intermediate, and high altitude populations with respect to the degree of CAM expression and the capability to tolerate limited water availability. We utilized clonallyreplicated genotypes of plants from each population in common environment greenhouse experiments. Genetic differences among the populations were found in long-term water use efficiency, in 24 hour CO2 exchange patterns, in biomass δ13C values, in carbon allocation, and in water status and ultimately survival during prolonged drought. The differences among the populations appear to be closely related to differences in the native habitats. The low altitude, desert plants had the greatest ability to grow and survive under conditions of limited water availability and appear to have the greatest shift to nighttime CO2 uptake during periods without water, while the high altitude plants had the poorest performance under these conditions and appear to shut down net carbon uptake when severely water limited.


Anterior cruciate ligament reconstruction: long-term results using autograft tissue.
Injuries involving the anterior cruciate ligament are quite common. Numerous surgical procedures have been proposed for the management of the symptomatic ACL-deficient knee. The most popular treatment modality is currently intra-articular reconstruction with the use of autogenous graft tissue. The focus of this article is to review the long-term results of autograft ACL reconstruction.


Serological diagnosis of Toxoplasma gondii infection: Recommendations from the French National Reference Center for Toxoplasmosis.
Toxoplasmosis manifests no clinical signs in 80% of cases in immunocompetent patient, causing immunization characterized by the persistence of cysts, particularly in brain, muscles, and retina. Assessing the serological status, based on testing for serum toxoplasma IgG and IgM antibodies, is essential in cases that are increasingly at risk for the more severe disease forms, such as congenital or ocular toxoplasmosis. This disease also exposes immunosuppressed patients to reactivation, which can lead to more widespread forms and increased mortality. By interpreting the serological results, we can estimate the risk of contamination or reactivation and define appropriate prophylactic and preventive measures, such as hygienic and dietetic, therapeutic, biological, and clinical follow-up, according to the clinical context. We hereby propose practical approaches based on serological data, resulting from a consensus of a group of experts from the French National Reference Center Network for Toxoplasmosis, according to both routine and specific clinical situations.


Cell-free technologies for biopharmaceutical research and production.
Cell-free protein synthesis (CFPS) technologies have grown from lab-scale research tools to biopharmaceutical production at the Good Manufacturing Practice manufacturing scale. Multiple human clinical trials are in progress with CFPS-based products. In addition, applications of CFPS in research have continued to expand over the years and play an important role in biopharmaceutical product discovery and development. The unique, open nature of CFPS has enabled efficient non-natural amino acid (nnAA) incorporation into protein products, which expands the range of biotherapeutics that can be considered for novel treatments. The flexibility and speed of CFPS combined with novel nnAA capabilities are poised to open a new chapter in the continuing evolution of biotherapies.


Cost effectiveness of treatment for alcohol problems: findings of the randomised UK alcohol treatment trial (UKATT).
To compare the cost effectiveness of social behaviour and network therapy, a new treatment for alcohol problems, with that of the proved motivational enhancement therapy. Cost effectiveness analysis alongside a pragmatic randomised trial. Seven treatment sites around Birmingham, Cardiff, and Leeds. 742 clients with alcohol problems; 617 (83.2%) were interviewed at 12 months and full economic data were obtained on 608 (98.5% of 617). Main economic measures Quality adjusted life years (QALYs), costs of trial treatments, and consequences for public sector resources (health care, other alcohol treatment, social services, and criminal justice services). Both therapies saved about five times as much in expenditure on health, social, and criminal justice services as they cost. Neither net savings nor cost effectiveness differed significantly between the therapies, despite the average cost of social behaviour and network therapy (221 pounds sterling; 385 dollars; 320 euros) being significantly more than that of motivational enhancement therapy (129 pounds sterling). If a QALY were worth 30,000 pounds sterling, then the motivational therapy would have 58% chance of being more cost effective than the social therapy, and the social therapy would have 42% chance of being more cost effective than the motivational therapy. Participants reported highly significant reductions in drinking and associated problems and costs. The novel social behaviour and network therapy did not differ significantly in cost effectiveness from the proved motivational enhancement therapy.


Influence of foetal genotype on the follicle-stimulating hormone:luteinizing hormone ratio of pregnant mare serum gonadotrophin.
Rat testicular radioreceptor assays specific for FSH and LH were used to determine the FSH:LH ratio of PMSG produced by horse, donkey, mule and hinny conceptuses. Measurements of FSH and LH activities in PMSG produced both in vivo and in vitro by the four types of conceptuses showed that the genotype of the foetus markedly influences the FSH:LH ratio of PMSG. The FSH:LH ratio of PMSG produced by the horse conceptus was around unity whereas the ratio of PMSG produced by the donkey conceptus was as low as 0-2. Furthermore, the hybrid mule and hinny conceptuses both produced PMSG with an FSH:LH ratio which was approximately midway between those of the horse and donkey.


A novel alternative splicing form of excitatory amino acid transporter 1 is a negative regulator of glutamate uptake.
Abstract EAAT1 is a major glutamate transporter in the CNS and is required for normal neurotransmission and neuroprotection from excitotoxicity. In the present study, we have identified a novel form of the human EAAT1, named here as EAAT1ex9skip, which lacks the entire exon 9. Quantitative PCR analysis indicates that this variant is expressed throughout the CNS, both in grey matter and axonal tracts, at levels ranging between 10% and 20% of the full-length EAAT1 form. When expressed in HEK293 cells, EAAT1ex9skip mRNA is translated into a truncated protein localized in the endoplasmic reticulum. EAAT1ex9skip has no functional glutamate uptake activity but instead, exerts a dominant negative effect over full-length EAAT1 function. In turn, co-expression of full-length EAAT1 and EAAT1ex9skip variants reduces the insertion of the former into the plasma membrane. Together, these results indicate that the EAAT1ex9skip splice variant is a negative regulator of full-length EAAT1 function in the human brain.


Cardiolipin accumulation in the inner and outer membranes of Escherichia coli mutants defective in phosphatidylserine synthetase.
Mutants of Escherichia coli defective in phosphatidylserine synthetase (pss) make less phosphatidylethanolamine than normal cells, and they are temperature sensitive for growth. We have isolated a new mutant, designated RA2021, which is better than previously available strains in that the residual phosphatidylethanolamine level approaches 25% after 4 h at 42 degrees C. The total amount of phospholipid normalized to the density of the culture is about the same in RA2021 (pss-21) as in the isogenic wild-type RA2000 (pss(+)). Consequently, there is a net accumulation of polyglycerophosphatides in the mutant, particularly of cardiolipin. The addition of 10 to 20 mM MgCl(2) to a culture of RA2021 prolongs growth under nonpermissive conditions and prevents loss of cell viability, but it does not eliminate the temperature-sensitive phenotype. Divalent cations, like Mg(2+), do not correct the phospholipid composition of the mutant, but may act indirectly by balancing the negative charges of phosphatidylglycerol and cardiolipin. To determine the effects of the pss mutation on membrane composition, we have examined the subcellular distribution of the polyglycerophosphatides that accumulate in these strains. All of the excess anionic lipids of RA2021 are associated with the envelope fraction and are distributed equally between the inner and outer membranes. The protein compositions of the isolated membranes do not differ significantly in the mutant and wild type. The fatty acid composition of RA2021 is almost the same as wild type at 30 degrees C, but there is more palmitic and cyclopropane fatty acid at 42 degrees C. These results demonstrate that the modification of the polar lipid composition observed in pss mutants affects both membranes and that cardiolipin, which is not ordinarily present in large quantities, can accumulate in the outer membrane when it is overproduced by the cell. The altered polar headgroup composition of the outer membrane in pss mutants may account, in part, for their hypersensitivity to the aminoglycoside antibiotics.


Sulfur K-edge X-ray absorption spectroscopy for determining the chemical speciation of sulfur in biological systems.
Sulfur is an essential biological element, yet its biochemistry is only partially understood because there are so few tools for studying this element in biological systems. X-ray absorption spectroscopy provides a unique approach to determining the chemical speciation of sulfur in intact biological samples. Different biologically relevant sulfur compounds show distinctly different sulfur K-edge X-ray absorption spectra, and we show here, as an example, that this allows the deconvolution of the sulfur species in equine blood.


Selective destruction of mossy fibers and granule cells with preservation of the GABAergic network in the inferior region of the rat hippocampus after colchicine treatment.
Lesions induced by colchicine injection into the rat hippocampus were investigated by means of electron microscopy and GABA immunocytochemistry. Granule cells were nearly completely destroyed 3 days after colchicine injection; since the necrosis of their axonal endings was delayed, an anterograde degeneration of the mossy fibers had probably taken place. The selectivity of the lesions was not limited to granule cells, for some pyramidal neurons in CA1 pyramidal layer were damaged. It was, however, striking to observe that throughout the hippocampal structure GABAergic neurons were spared from the effects of colchicine. For instance, GABAergic neurons were found in the vicinity of the completely destroyed granule cell layer. GABAergic neurons and terminals were also present in the CA3 region where the GABA-containing terminals formed a dense network of synapses with somata and dendrites of pyramidal cells. It was interesting to note that, consistent with previous studies, the GABAergic neurons in CA3 are innervated by mossy fibers. We conclude that after colchicine treatment the destruction of the granule cells was not associated with a lesion of the GABAergic network. This selective lesion provides a useful model with which to study the properties of CA3 neurons deprived of their major excitatory input but with an intact inhibitory network.


Chiral derivatizing agents, macrocycles, metal complexes, and liquid crystals for enantiomer differentiation in NMR spectroscopy.
Enantiomerically pure chiral auxiliary agents are often used in NMR spectroscopy to facilitate the differentiation of enantiomers. Chiral derivatizing agents are covalently bound to the substrate and differences in chemical shifts of the resulting diastereomeric complexes are used in the analysis. Macrocycles such as cyclodextrins, crown ethers, and calix[4]resorcinarenes are chiral solvating agents that associate with the substrate through non-covalent interactions. Enantiomeric differentiation occurs in the NMR spectrum because of the diastereomeric nature of the associated complexes and/or because of the differences in association constants between the two enantiomers and the chiral reagent. Metal complexes are Lewis acids that bind to suitable Lewis base donor compounds. Exchange of substrate can be slow or fast depending on the particular metal ion, mimicking the behavior of a chiral derivatizing or solvating agent, respectively. Chiral liquid crystals undergo a partial alignment in an applied magnetic field and enantiomers dissolved in the liquid crystal undergo a partial alignment as well. If the alignment of the two enantiomers is different, enantiomeric differentiation can potentially be observed by differences in chemical shifts, differences in dipolar coupling constants, and different magnitudes of splitting of quadrupolar nuclei such as deuterium. The chiral reagents described herein can be used to determine enantiomeric composition and sometimes to assign absolute configuration. Significant discoveries as well as recent findings with each of these types of systems are described.


The association of bisphenol A and paraben exposure with sensorineural hearing loss in children.
Bisphenol A (BPA) and parabens (PBs) are chemicals that are extensively used in personal care products (PCPs). In early childhood development, hearing is critical to speech and language development, communication, and learning. In vitro and in vivo, BPA/PBs exhibited neurotoxicity through elevated levels of oxidative stress. BPA also has the potential to be an ototoxicant. Therefore, this study aimed to determine the association of exposure to BPA/PBs with sensorineural hearing loss in children. A cross-sectional study based on hearing tests was conducted. This study enrolled 320 children aged 6-12 years from elementary school. Urinary BPA and PB concentrations were analyzed by using liquid chromatography-tandem mass spectrometry (LC-MS/MS). Logistic regression models were employed to determine the association of BPA/PB exposure with sensorineural hearing loss. Children with sensorineural hearing loss had higher BPA concentrations than normal-hearing children (0.22 ng/ml vs. 0.10 ng/ml, p = 0.05). After adjustment for covariates, the risk of hearing loss at middle frequencies reached 1.83-fold (95% CI: 1.12-2.99) when BPA concentrations increased by 1 log10. The risk of slight hearing loss reached 2.24-fold (95% CI: 1.05-4.78) when children had a tenfold increase in ethyl paraben (EP) concentration. This study clarifies the role of exposure to BPA/PBs in hearing loss in children. Future research needs to be expanded to include cohort designs and nationwide studies to identify causality.


Effect of ibudilast on thalamic magnetization transfer ratio and volume in progressive multiple sclerosis.
Thalamic volume (TV) is a sensitive biomarker of disease burden of injury in multiple sclerosis (MS) and appears to reflect overall lesion loads. Ibudilast showed significant treatment effect on brain atrophy and magnetization transfer ratio (MTR) of normal-appearing brain tissue but not in new/enlarging T2 lesion in the SPRINT-MS randomized clinical trial. To evaluate the effect of ibudilast on thalamic tissue integrity and volume in the SPRINT-MS. A total of 255 participants with progressive MS were randomized to oral ibudilast or placebo, and thalamic MTR and normalized TV over 96 weeks were quantified. Mixed-effect modeling assessed treatment effects on the thalamic MTR and TV, separately. Similarly, the measures were compared between the participants with confirmed disability progression (CDP). Ibudilast's treatment effect was observed compared to placebo for thalamic MTR (p = 0.03) but not for TV (p = 0.68) while TV correlated with T2 lesion volume (p < 0.001). CDP associated with thalamic MTR (p = 0.04) but not with TV (p = 0.7). Ibudilast showed an effect on thalamic MTR, which was associated with CDP, suggesting a clinically relevant effect on thalamic tissue integrity. However, the treatment effect was not observed in TV, suggesting that thalamic atrophy is more closely associated with global inflammatory activity than local tissue integrity. NCT01982942.


Adiponectin, leptin, and leptin/adiponectin ratio with risk of gestational diabetes mellitus: A prospective nested case-control study among Chinese women.
To examine the associations of serum concentrations of adiponectin and leptin and leptin/adiponectin ratio (LAR) in early pregnancy with risk of gestational diabetes mellitus (GDM) in Chinese women. The predictive ability of those biomarkers for GDM was also assessed. Within the Tongji-Shuangliu Birth Cohort, a nested case-control study was established with 332 GDM cases and 664 matched controls at 1:2 ratio on age (±3 years) and gestational age (±4 weeks). Serum adiponectin and leptin levels were measured in early pregnancy (median gestational week, 11; range, 6-15). Conditional logistic regression models with adjustment for potential covariates were used to evaluate the associations. Multivariable-adjusted odds ratios (ORs) comparing extreme quartiles of adiponectin, leptin and LAR were 0.55 (95 % CI, 0.35, 0.85), 1.96 (95 % CI, 1.19, 3.24), and 2.72 (95 % CI, 1.63, 4.54) for GDM, respectively (All P-trend < 0.02). Adding adiponectin and leptin to a conventional prediction model (including traditional risk factors and fasting glucose) increased the C-statistics from 0.708 (95 % CI, 0.674, 0.741) to 0.728 (95 % CI, 0.695, 0.760), and achieved a net reclassification improvement of 0.292. Our findings indicate that adiponectin is inversely associated with GDM, while leptin and LAR are positively associated with GDM in Chinese pregnant women.


Successful outcome with tandem myeloablative chemotherapy and autologous peripheral blood stem cell transplants in a patient with atypical teratoid/rhabdoid tumor of the central nervous system.
Atypical teratoid rhabdoid tumors (ATRT) are highly malignant tumors of the central nervous system with a peak incidence in children less than 3 years of age. Despite multimodal therapy including surgery, radiation and chemotherapy, the prognosis remains dismal. No specific treatment guidelines are defined for ATRTs but a gross total resection and radiation therapy (RT) appear to improve overall outcome. In children less than 3 years of age, the prognosis is dismal due in part to the reluctance to utilize RT given its severe neurological sequelae. To avoid RT in this age group, intensification of chemotherapy has been tried and has shown to improve outcome. Myeloablative chemotherapy followed by autologous stem cell re-infusion has been used as a modality to intensify therapy but there are no reports of use of tandem myeloablative regimens and autologous stem cell re-infusions for treatment of ATRT. We herein report the case of a 4-month-old boy with ATRT with partial resection of his tumor who achieved complete remission using tandem high-dose therapy followed by autologous peripheral blood stem cell re-infusions despite having biopsy proven disease at the time of starting the tandem regimens. This was achieved without the use of RT as a treatment modality.


A review of psychocutaneous disorders from a psychotherapeutic perspective-Toolkit for the dermatologist.
The study of psychocutaneous disorders requires a multidisciplinary approach. It is of paramount importance that dermatologists understand the psychiatric premise for these disorders. Mental health professionals can also benefit from a better understanding of the dermatologic manifestations of psychiatric disorders they may encounter in their practice. The aim of this study is to elevate the level of understanding regarding psychotherapeutic treatment of psychocutaneous disorders. We performed a literature review using the major databases. Four researchers reviewed English, full text, peer-reviewed articles that were published after 2000 using our specific search terms and inclusion/exclusion criteria. The majority of psychocutaneous disorders seem to be clustered among three DSM-5 disorders: depressive disorders, anxiety disorders, and obsessive-compulsive related disorders. Better recognition of the underlying psychiatric comorbidities may lead to improved patient outcomes.


Evaluation of articles on pulmonology published in Brazilian journals other than the Brazilian Journal of Pulmonology.
In Brazil, research on pulmonology has become increasingly more visible in recent years. In addition to the Brazilian Journal of Pulmonology, other journals have contributed to that by publishing relevant articles in this area. The objective of this article was to briefly report the most relevant studies on pulmonology that were published in other important Brazilian journals between 2009 and 2010. Altogether, there were 56 articles related to the various subareas that compose the field of respiratory diseases.


Radiation exposure from medical imaging in dialyzed patients undergoing renal pre-transplant evaluation.
Ionizing radiation exposure from medical procedures is rising sharply-the per-capita annual effective dose in the US is 3.0 millisieverts (mSv). Hemodialyzed and kidney transplanted patients receive still higher doses of ionizing radiation due to the presence of multiple comorbidities. The aim of this study was to assess the cumulative effective dose (CED) among dialyzed patients undergoing renal pre-transplant evaluation. We evaluated 70 hemodialysis patients between June 2009 and December 2014, aged 46.4 ± 12.0 years. The number and type of radiologic procedures were collected through the Radiology Information System. CED was expressed as total mSv/patient and annual CED (mSv/patient/year). A total of 744 radiologic procedures were performed, accounting for 3869 mSv of ionizing radiation: conventional radiology, computed tomography and nuclear medicine accounted for 78, 14 and 8 % of the procedures, but they represented, respectively, 8, 83 and 9 % of the total CED. The mean (median) annual CED was 35 (7) mSv/patient/year, while total CED was 72 (32) mSv/patient. Thirty-seven patients were active waitlisted and received 47 (10) mSv during the pre-transplant evaluation and 36 (5) mSv during the waiting phase to maintain active status. Concerning cancer risk, 4 (7 %) patients were classified at low risk (<3 mSv/year), 19 (35 %) at moderate risk (3 to <20 mSv/year), 8 (15 %) at high risk (20 to <50 mSv/year), and 23 (43 %) at very high risk (≥50 mSv/year). Our study demonstrated that during renal pre-transplant evaluation, dialyzed patients receive a high dose of ionizing radiation. Considering that transplanted individuals have a high incidence of cancer due to multifactorial etiology, it is mandatory to reduce the ionizing radiation imaging.


Review article: small intestinal bacterial overgrowth--prevalence, clinical features, current and developing diagnostic tests, and treatment.
The symptoms and signs of small intestinal bacterial overgrowth (SIBO) are often identical to a variety of diseases and can lead to diagnostic confusion. To review the diagnostic options for SIBO and present new investigative options for the condition. A literature search was performed on MEDLINE, EMBASE and Web of Science for English articles and abstracts. Search terms included free text words and combinations of the following terms 'small intestinal bacterial overgrowth', 'small bowel bacterial overgrowth', 'diagnostic tests', 'treatment', 'antibiotics', 'probiotics', 'metabonomics', 'proton nuclear magnetic resonance spectroscopy', 'electronic nose' and 'field asymmetric ion mobility spectrometry'. All of the available methods to test for SIBO have inherent limitations and no 'gold-standard' diagnostic test for the condition exists. Accurate diagnosis of SIBO requires identification of bacterial species growing inappropriately within the small intestine and symptom response to antibiotics. Proton nuclear magnetic resonance spectroscopy, electronic nose technology and/or field asymmetric ion mobility spectrometry may represent better investigative options for the condition. Novel diagnostic options are needed to supplement or replace available tests.


Detection of ABCC2, CYP2B6 and CYP3A4 in human peripheral blood mononuclear cells using flow cytometry.
ABCC2 has a wide tissue distribution and can mediate the efflux of a number of therapeutic compounds from cells and contribute to potential treatment failure. Its diverse expression and ability to efflux a number of substrates imply a number of physiological and pharmacological roles. CYP2B6 and CYP3A4 are responsible for the metabolism of a number of therapeutic compounds. Reports on the expression of these proteins in various cells and tissues have been contradictory mainly due to differences in experimental approach and cell type studied. With the advances in commercially available antibodies we describe here a simplified technique for the detection of ABCC2, CYP2B6 and CYP3A4 in human peripheral blood mononuclear cells (PBMC) by flow cytometry. Results are expressed as mean increase in fluorescence compared to isotypically matched controls. Using these assays we confirmed the expression of these proteins in human PBMC. These methods are rapid and reproducible and have potential use for both in vitro and clinical applications.


Pelvic exsanguination following umbilical artery catheterization in neonates.
Umbilical artery catheterization in 400 newborns was followed by umbilical or hypogastric artery perforation with massive extra- and intraperitoneal bleeding in 7 infants. Common predisposing factors were multiple attempts and insertion against resistance. A characteristic clinical picture developed, including hypotension, abdominal distention, and lower abdominal wall induration and ecchymosis, often extending into the genitalia and upper thighs, with absent femoral pulses. Treatment is immediate transfusion and bilateral umbilical and hypogastric artery ligation. One of seven patients survived.


Successful treatment with voriconazole of fungal cerebral abscess due to Cladophialophora bantiana.
We report the first successfully treated case of Cladophialophora bantiana cerebral abscess with the relatively new antifungal agent voriconazole. Infection with this organism is often fatal. A 64-year-old man presented to our institution with progressive neurologic symptoms due to a brain abscess. A stereotactic brain biopsy confirmed the pathogen as C. bantiana. We discuss the successful treatment of this patient, and review the pharmacological actions of voriconazole and the literature on the treatment of this organism. Previously considered a rare cause of cerebral abscess, C. bantiana fungal infections have become more common in recent years. Aggressive and continuous treatment with voriconazole may offer an improved chance of survival in these patients.


Count in Time So All is Fine.
Dislodgement of surgical sponge into airway during the intraoperative period is uncommon as the airway, in most cases secured by an endotracheal tube. We report such an unusual case during micro laryngeal surgery and direct laryngoscopy assessment under general anaesthesia. This shows early suspicion and quick action to avoid disaster.


Annular pancreas detected in adults, diagnosed by endoscopic retrograde cholangiopancreatography: report of four cases.
Four cases of annular pancreas diagnosed by endoscopic retrograde cholangiopancreatography (ERCP) are described and 105 cases of this anomaly in adults in Japan were reviewed. Among 105 cases, abdominal pain was the most frequent symptom. Concerning associated diseases, peptic ulcer was present in 24.8% and pancreatitis in 13.3%. In case 1, duodenal ulcer and pancreatic cyst were noted. Pancreatolithiasis was found in two cases (case 1 and 2). Case 4 presented the clinical features of acute pancreatitis. Out of 105 cases, well-described 26 were divided into six types. The following results were obtained. 1) The most frequent type was that in which the annular duct arose from the duct of Wirsung. 2) The next most frequent type was that in which the main pancreatic duct encircled the duodenum. 3) The other types corresponded to those in which the annular duct arose from the duct of Santorini and the common bile duct. We emphasized that ERCP is the most important procedure to find the characteristic features and to establish the therapeutic strategy in cases of annular pancreas.


[Pyogenic sacroiliitis in a patient with HLA B27].
The authors report on a patient with bilateral sacroillitis and positive HLA B27 who developed a pyoarthritis in one of the sacro-iliac joints. They comment the diagnostics difficulties in this type of septic arthritis and the association of the histocompatibility antigen B27 with pyogenic sacroiliitis.


Notoginsenoside R1 stimulates osteogenic function in primary osteoblasts via estrogen receptor signaling.
Notoginsenoside R1 (NGR1), a novel phytoestrogen isolated from Panax notoginseng, has been widely used in the treatment of microcirculatory diseases in Asian countries. Here we investigated the effect of NGR1 on osteoblast differentiation and mineralization process. Furthermore, we also evaluated NGR1's estrogenic properties, especially its effects on estrogen receptors (ERs). NGR1 activated the transcriptional activity of phosphorylated estrogen response element (pERE)-luciferase (Luc) and induced ERα phosphorylation in hBMSC. In addition, ER activation correlated with induction and was associated with osteoblast differentiation biomarkers including alkaline phosphatase activity and transcription of osteoblastic genes, e.g., type I collagen (COL1), osteonectin, osteocalcin (OC), runt related protein 2 (Runx2), and osterix. NGR1 also promoted the mineralization process of osteoblasts. The NGR1-induced effects were confirmed to be mediated by the ER by the observation that pretreatment of the osteoblasts with the ER antagonist, ICI 182,780 fully blocked the effects. Our results showed that NGR1 stimulates osteogenic differentiation of cultured osteoblasts by activating ER signaling and in turn might be a potential therapeutic alternative for the prevention and treatment of osteoporosis.


Do Estimates of Treatment Risk Based on Clinical Vignettes Differ by Physician Gender?
Clinical vignettes are frequently used as instructional and evaluative instruments for physicians. Physicians' gender is a source of unconscious bias in treatment recommendations. This study assessed whether interpretation of information in clinical vignettes differed by physicians' gender as a possible source of unconscious bias. Thoracic surgeons and physicians in cardiothoracic surgical training were asked to provide estimates of major complication rates for lung resection on the basis of anonymized clinical vignettes of patients undergoing lung resection. Vignettes were categorized as low, average, and high risk by using a sum of Charlson Comorbidity Index (possible range, 0 to 37) and a combined physiologic score, EVAD (forced expiratory volume in 1 second, diffusing capacity of lung for carbon monoxide, age; possible range, 0 to 12); participants were not aware of the risk scores or vignette categories. Generalized estimating equation linear regression models were fit with risk scores treated as a continuous independent variable. A total of 247 physicians (105 practicing surgeons, 142 trainees; 203 men, 44 women) participated in one or more of the studies. Nearly all (103; 98%) of the practicing surgeons rated themselves as competent or expert in lung resection compared with 77 (54%) of the trainees (p < 0.001). Participants' complication estimates mirrored both vignette risk category and combined risk score. There was no significant difference between men and women physicians in their estimates of complication rates. Unconscious bias related to physicians' gender is not associated with differential use of information in clinical vignettes. Any possible bias may arise from face-to-face interactions with patients. Research into physicians' and patients' gender differences during such interactions is warranted.


A comparison of thermal injury, healing patterns, and postoperative adhesion formation following CO2 laser and electromicrosurgery.
To further define the role of CO2 laser in infertility surgery, the authors compared the immediate tissue damage and the subsequent healing patterns that followed the randomized use of CO2 laser and electrocautery on opposite sides of the reproductive tracts of 20 mature, female rabbits. Blind histopathologic evaluations were performed on the ovarian and uterine tissues immediately to compare the extent of the acute thermal damage, and 4 weeks postoperatively to compare the healing patterns. There were no differences in the depth of thermal damage, extent of collagen deposition, or in postoperative adhesion formation between CO2 laser and electrocautery. However, the mean depth of the acute thermal damage was significantly less on the ovary than on the uterus, and the area of fibrosis was significantly less when the incised uterine surfaces were approximated.


A long surviving case of resected gastric cancer presenting with a synchronous adrenal metastasis.
Clinically curable adrenal metastases are rare. We treated a patient with gastric cancer and a synchronous adrenal metastasis who underwent curative resection. Upper GI examinations of a 75-year-old man revealed a Borrmann 3 gastric tumor in the proximal stomach. CT indicated a giant gastric tumor that invaded the pancreatic tail, and the left adrenal gland seemed normal. He was subjected to a total gastrectomy and a distal pancreatosplenectomy. Because a mass was palpated intraoperatively in the left adrenal gland, it was also removed. The gastric tumor was histopathologically a poorly differentiated adenocarcinoma with scirrhous invasion that invaded the pancreas, and the histopathological findings of the left adrenal tumor were compatible with those of gastric cancer. At present, 6 years after the operation, there has been no clear sign of cancer recurrence. It may be rational to excise the left adrenal gland en bloc in patients with serosa-positive Borrmann 3-4 gastric cancer.


Effect of Recombinant Tissue Plasminogen Activator and 120-kHz Ultrasound on Porcine Intracranial Thrombus Density.
Surgical intervention for the treatment of intracerebral hemorrhage (ICH) has been limited by inadequate lysis of the target thrombus. Adjuvant transcranial ultrasound exposure is hypothesized to improve thrombolysis, expedite hematoma evacuation and improve clinical outcomes. A juvenile porcine intracerebral hemorrhage model was established by direct infusion of autologous blood into the porcine white matter. Thrombi were either not treated (sham) or treated with recombinant tissue plasminogen activator alone (rt-PA only) or in combination with pulsed transcranial 120-kHz ultrasound (sonothrombolysis). After treatment, pigs were euthanized, the heads frozen and sectioned and the thrombi extracted. D-Dimer and thrombus density assays were used to assess degree of lysis. Both porcine and human D-dimer assays tested did not have sufficient sensitivity to detect porcine D-dimer. Thrombi treated with rt-PA with or without 120-kHz ultrasound had a significantly lower density compared with sham-treated thrombi. No enhancement of rt-PA-mediated thrombolysis was noted with the addition of 120-kHz ultrasound (sonothrombolysis). The thrombus density assay revealed thrombolytic efficacy caused by rt-PA in an in vivo juvenile porcine model of intracerebral hemorrhage. Transcranial sonothrombolysis did not enhance rt-PA-induced thrombolysis, likely because of the lack of exogenous cavitation nuclei.


The efficacy of amineptine in the treatment of depressive patients with irritable bowel syndrome.
Patients with irritable bowel syndrome (IBS) often suffer from depression. In view of this, the effect of amineptine on the psychopathological condition of depressive patients with IBS was studied. Forty patients who satisfied the criteria for irritable bowel syndrome and had a Hamilton 24-item score above 15 were randomly assigned to receive either amineptine 200 mg/day or placebo in a double-blind clinical trial. Patients on amineptine were more improved at the end of the trial than patients on placebo (total Hamilton score). Amineptine was more effective on depressive mood, retardation, and cognitive dysfunction. Although these findings should be interpreted with caution because the baseline scores were higher in the amineptine than in the placebo group, they provide some evidence that amineptine may be a useful tool for the management of depressive patients with IBS.


Multivariate analysis of structural parameters of the optic nerve head assessed by means of confocal scanning laser (Heidelberg Retina Tomograph II).
We evaluated the optic nerve head parameters measured by confocal scanning laser that discriminated between normal and glaucomatous subjects in 97 normal eyes and 94 glaucomatous eyes. Several different linear regression formulas can discriminate the presence of glaucomatous. The use of linear regression functions of HRT parameters elaborated from autochthon population around the world will add the amounts of evidence needed to support the global use of these functions.


MicroRNAs as pharmacological targets in endothelial cell function and dysfunction.
Endothelial cell dysfunction is a term which implies the dysregulation of normal endothelial cell functions, including impairment of the barrier functions, control of vascular tone, disturbance of proliferative, migratory and morphogenic capacities of endothelial cells, as well as control of leukocyte trafficking. MicroRNAs are short non-coding RNAs that have emerged as critical regulators of gene expression acting predominantly at the post-transcriptional level. This review summarizes the latest insights in the identification of endothelial-specific microRNAs and their targets, as well as their roles in controlling endothelial cell functions in both autocrine and paracrine manner. In addition, we discuss the therapeutic potential for the treatment of endothelial cell dysfunction and associated vascular pathophysiological conditions.


The contribution of prenatal sonographic diagnosis of clubfoot to preventive medicine.
From 1995 through 1997, clubfoot was detected by transabdominal sonography in 13 fetuses of 12 women, in nine bilaterally and in four unilaterally (total, 22 clubfeet). The average menstrual age at diagnosis was 23.6 weeks (range, 17-36). Three fetuses had associated malformations: two were therapeutically aborted and one died 2 weeks after birth. Proper understanding of the significance of prenatal ultrasound findings of clubfoot will assist both the orthopaedist and the parents in reaching a decision concerning future management of their pregnancy.


Mechanism of multi-site phosphorylation from a ROCK-I:RhoE complex structure.
The ROCK-I serine/threonine protein kinase mediates the effects of RhoA to promote the formation of actin stress fibres and integrin-based focal adhesions. ROCK-I phosphorylates the unconventional G-protein RhoE on multiple N- and C-terminal sites. These phosphorylation events stabilise RhoE, which functions to antagonise RhoA-induced stress fibre assembly. Here, we provide a molecular explanation for multi-site phosphorylation of RhoE from the crystal structure of RhoE in complex with the ROCK-I kinase domain. RhoE interacts with the C-lobe alphaG helix of ROCK-I by means of a novel binding site remote from its effector region, positioning its N and C termini proximal to the ROCK-I catalytic site. Disruption of the ROCK-I:RhoE interface abolishes RhoE phosphorylation, but has no effect on the ability of RhoE to disassemble stress fibres. In contrast, mutation of the RhoE effector region attenuates RhoE-mediated disruption of the actin cytoskeleton, indicating that RhoE exerts its inhibitory effects on ROCK-I through protein(s) binding to its effector region. We propose that ROCK-I phosphorylation of RhoE forms part of a feedback loop to regulate RhoA signalling.


Loss of hand and forearm following Bier block: a case report.
Few complications have been reported with the use of the Bier block for intravenous regional anesthesia. The method requires little expert training and produces good analgesia in a bloodless field for a period of 1 to 1.5 hours. The loss of an upper extremity in a 25-year-old white woman who had the performance of a Bier block for removal of a ganglion cyst is reported. The block was performed with 200 mg of mepivacaine without epinephrine. Total tourniquet time was 25 minutes. The patient had prompt onset of pain and swelling followed by loss of motor and sensory function. Operative exploration revealed necrosis of the forearm compartments. Arteriography demonstrated thrombosis of the terminal branches of the radial and ulnar artery. Below elbow amputation was performed 1 week later. Essentially all complications reported with regional anesthesia have been related to the systemic pharmacologic effects of the local agent used. Loss of an upper extremity has not been reported. We postulate three possible mechanisms: (1) possible cannulation and injection of the radial artery, (2) an idiosyncratic allergic reaction to the anesthetic agent or the preservative, and (3) erroneous injection of a foreign substance into the forearm venous system. We conclude the last mechanism is the most likely cause in this patient.


A novel lipid compound, epolactaene, induces apoptosis: its action is modulated by its side chain structure.
A novel lipid compound, epolactaene, was isolated from the culture supernatant of Penicillium sp. 1689-P and it has already been reported that it induced neurite outgrowth in a human neuroblastoma cell line. In this study, we first investigated the effects of epolactaene on a human leukemia B-cell line, BALL-1 cells, and clarified that epolactaene induces apoptosis in BALL-1 cells in a dose- and time-dependent manner. Furthermore, we focused on the side chain structure of epolactaene, and chemically synthesized epolactaene derivatives. One derivative, which has a straight long alkyl chain as its side chain, induced apoptosis more effectively than epolactaene. On the other hand, other derivatives with a short alkyl side chain had weaker apoptosis-inducing actions. A good correlation was found between the apoptosis-inducing action of these compounds and their octanol/water partition coefficients (log P). These results suggested that the apoptosis-inducing activities of epolactaene and its derivatives were related to the hydrophobicity of these compounds; so that side chain structure of epolactaene is very important for its apoptosis-inducing activities. These apoptosis-inducing actions of epolactaene and its derivatives were also observed in various blood tumor cell lines and normal lymphocytes.


Bone Tissue Engineering.
Medical advances have led to a welcome increase in life expectancy. However, accompanying longevity introduces new challenges: increases in age-related diseases and associated reductions in quality of life. The loss of skeletal tissue that can accompany trauma, injury, disease or advancing years can result in significant morbidity and significant socio-economic cost and emphasise the need for new, more reliable skeletal regeneration strategies. To address the unmet need for bone augmentation, tissue engineering and regenerative medicine have come to the fore in recent years with new approaches for de novo skeletal tissue formation. Typically, these approaches seek to harness stem cells, innovative scaffolds and biological factors that promise enhanced and more reliable bone formation strategies to improve the quality of life for many. This review provides an overview of recent developments in bone tissue engineering focusing on skeletal stem cells, vascular development, bone formation and the translation from preclinical in vivo models to clinical delivery.


Prognostic role of cardiac power index in ambulatory patients with advanced heart failure.
Cardiac pump function is often quantified by left ventricular ejection fraction by various imaging modalities. As the heart is commonly conceptualized as a hydraulic pump, cardiac power describes the hydraulic function of the heart. We aim to describe the prognostic value of resting cardiac power index (CPI) in ambulatory patients with advanced heart failure. We calculated CPI in 495 sequential ambulatory patients with advanced heart failure who underwent invasive haemodynamic assessment with longitudinal follow-up of adverse outcomes (all-cause mortality, cardiac transplantation, or ventricular assist device placement). The median CPI was 0.44 W/m(2) (interquartile range 0.37, 0.52). Over a median of 3.3 years, there were 117 deaths, 104 transplants, and 20 ventricular assist device placements in our cohort. Diminished CPI (<0.44 W/m(2) ) was associated with increased adverse outcomes [hazard ratio (HR) 2.4, 95% confidence interval (CI) 1.8-3.1, P < 0.0001). The prognostic value of CPI remained significant after adjustment for age, gender, pulmonary capillary wedge pressure, cardiac index, pulmonary vascular resistance, left ventricular ejection fraction, and creatinine [HR 1.5, 95% CI 1.03-2.3, P = 0.04). Furthermore, CPI can risk stratify independently of peak oxygen consumption (HR 2.2, 95% CI 1.4-3.4, P = 0.0003). Resting cardiac power index provides independent and incremental prediction in adverse outcomes beyond traditional haemodynamic and cardio-renal risk factors.


Antibiotics and antibiotic resistance genes in anaerobic digesters and predicted concentrations in agroecosystems.
In recent decades, the innovative practice of management and valorization of agrozootechnical waste as energy through anaerobic digestion (AD) has been rapidly growing. However, whether applying digestate to soil as biofertilizer can be a source of antibiotics (ABs) and antibiotic resistance genes (ARGs) has not been fully investigated so far. In this work the ARGs responsible for sulfamethoxazole (SMX) resistance (sul1, sul2), ciprofloxacin (CIP) resistance (qnrS, qepA, aac-(6')-Ib-cr) and the mobile genetic element intl1, together with the concentrations of the antibiotics SMX and CIP, were measured in several anaerobic digesters located in Central Italy. Based on these results, the concentrations of antibiotics and ARGs which can potentially reach soil through amendment with digestate were also estimated. The highest CIP and SMX concentrations were found during winter and spring in anaerobic digesters. The highest ARG abundances were found for the aac-(6')-Ib-cr and sul2 genes. The overall results showed that application of digestate to soil does not exclude AB contamination and spread of ARGs in agroecosystems, especially in the case of ciprofloxacin, owing to its high intrinsic persistence.


Genome Modification of Pluripotent Cells by Using Transcription Activator-Like Effector Nucleases (TALENs).
Interest is increasing in transcription activator-like effector nucleases (TALENs) as a tool to introduce targeted double-strand breaks into the large genomes of human and animal cell lines. The produced DNA lesions stimulate DNA repair pathways, error-prone but dominant non-homologous end joining (NHEJ) and accurate but less occurring homology-directed repair (HDR), and as a result targeted genes can be modified. Here, we describe a modified Golden-Gate cloning method for generating TALENs and also details for targeting genes in mouse embryonic stem cells. The protocol described here can be used for modifying the genome of a broad range of pluripotent cell lines.


Cytotoxicity of silica-glass fiber reinforced composites.
Silica-glass fiber reinforced polymers can be used for many kinds of dental applications. The fiber reinforcement enhances the mechanical properties of the polymers, and they have good esthetic attributes. There is good initial bonding of glass fibers to polymers via an interface made from silane coupling agents. The aim of this in vitro study was to determine the cytotoxicity of two polymers reinforced with two differently sized silica-glass fibers before and after thermal cycling. Cytotoxicity of the polymers without fibers was also evaluated. Two different resin mixtures (A and B) were prepared from poly(vinyl chloridecovinylacetate) powder and poly(methyl methacrylate) (PMMA) dissolved in methyl methacrylate and mixed with different cross-linking agents. The resin A contained the cross-linking agents ethylene glycol dimethacrylate and 1,4-butanediol dimethacrylate, and for resin B diethylene glycol dimethacrylate was used. Woven silica-glass fibers were used for reinforcement. The fibers were sized with either linear poly(butyl methacrylate)-sizing or cross-linking PMMA-sizing. Cytotoxicity was evaluated by filter diffusion test (ISO 7405:1997) of newly made and thermocycled test specimens. Extracts were prepared according to ISO 10993-12 from newly made and from thermocycled specimens and tested by the MTT assay. The results from the experiments were statistically analyzed by one-way ANOVA and Tukey's test (rho<0.05). The filter diffusion test disclosed no change in staining intensity at the cell-test sample contact area indicating non-cytotoxicity in all experimental groups. Cell viability assessed by MTT assay was more than 90% in all experimental groups. All are non-cytotoxic. It can be concluded that correctly processed heat polymerized silica-glass fiber reinforced polymers induced no cytotoxicity and that thermocycling did not alter this property.