Risks factors for infections with extended-spectrum beta-lactamase-producing Escherichia coli and Klebsiella pneumoniae at a tertiary care university hospital in Switzerland.
There are considerable geographical differences in the occurrence of extended-spectrum beta-lactamase(ESBL)-producing bacteria, both in the community and in the hospital setting. Our aim was to assess risk factors for blood stream, urinary tract, and vascular catheter-associated infections with ESBL-producing Escherichia coli and Klebsiella pneumoniae at a tertiary care hospital in a low-prevalence country. We performed a case-control study comparing 58 patients with infections due to ESBL-producing E. coli orK. pneumoniae vs 116 controls with infections due to non-ESBL producing organisms at the University Hospital Zurich, Switzerland, between 1 July 2005 and 30 June 2007. Cases included 15 outpatients and 43 inpatients. Multivariable analyses found three risk factors for ESBL-producing isolates: begin of symptoms or recent antibiotic pre-treatment in a foreign country (odds ratio [OR] 27.01,95% confidence interval [CI] 2.38-1,733.28], p = 0.042),antibiotic therapy within the year preceding the isolation of the ESBL-producing strain (OR 2.88, 95% CI 1.13-8.49,p = 0.025), and mechanical ventilation (OR 10.56, 95% CI 1.06-579.10, p = 0.042). The major risk factors for infections due to ESBL-producing bacteria were travel in high-prevalence countries, prior antibiotic use, and mechanical ventilation during a stay in the intensive care unit. Community-acquired infections were documented in 17% of the patients.An early identification of risk factors is crucial to providing the patients an optimal empiric antibiotic therapy and to keep the use of carbapenems to a minimum.


Genes encoding major light-harvesting polypeptides are clustered on the genome of the cyanobacterium Fremyella diplosiphon.
The polypeptide composition of the phycobilisome, the major light-harvesting complex of prokaryotic cyanobacteria and certain eukaryotic algae, can be modulated by different light qualities in cyanobacteria exhibiting chromatic adaptation. We have identified genomic fragments encoding a cluster of phycobilisome polypeptides (phycobiliproteins) from the chromatically adapting cyanobacterium Fremyella diplosiphon using previously characterized DNA fragments of phycobiliprotein genes from the eukaryotic alga Cyanophora paradoxa and from F. diplosiphon. Characterization of two lambda-EMBL3 clones containing overlapping genomic fragments indicates that three sets of phycobiliprotein genes--the alpha- and beta-allophycocyanin genes plus two sets of alpha- and beta-phycocyanin genes--are clustered within 13 kilobases on the cyanobacterial genome and transcribed off the same strand. The gene order (alpha-allophycocyanin followed by beta-allophycocyanin and beta-phycocyanin followed by alpha-phycocyanin) appears to be a conserved arrangement found previously in a eukaryotic alga and another cyanobacterium. We have reported that one set of phycocyanin genes is transcribed as two abundant red light-induced mRNAs (1600 and 3800 bases). We now present data showing that the allophycocyanin genes and a second set of phycocyanin genes are transcribed into major mRNAs of 1400 and 1600 bases, respectively. These transcripts are present in RNA isolated from cultures grown in red and green light, although lower levels of the 1600-base phycocyanin transcript are present in cells grown in green light. Furthermore, a larger transcript of 1750 bases hybridizes to the allophycocyanin genes and may be a precursor to the 1400-base species.


Aluminum-Activated Malate Transporters Can Facilitate GABA Transport.
Plant aluminum-activated malate transporters (ALMTs) are currently classified as anion channels; they are also known to be regulated by diverse signals, leading to a range of physiological responses. Gamma-aminobutyric acid (GABA) regulation of anion flux through ALMT proteins requires a specific amino acid motif in ALMTs that shares similarity with a GABA binding site in mammalian GABAA receptors. Here, we explore why TaALMT1 activation leads to a negative correlation between malate efflux and endogenous GABA concentrations ([GABA]i) in both wheat (Triticum aestivum) root tips and in heterologous expression systems. We show that TaALMT1 activation reduces [GABA]i because TaALMT1 facilitates GABA efflux but GABA does not complex Al3+ TaALMT1 also leads to GABA transport into cells, demonstrated by a yeast complementation assay and via 14C-GABA uptake into TaALMT1-expressing Xenopus laevis oocytes; this was found to be a general feature of all ALMTs we examined. Mutation of the GABA motif (TaALMT1F213C) prevented both GABA influx and efflux, and resulted in no correlation between malate efflux and [GABA]i We conclude that ALMTs are likely to act as both GABA and anion transporters in planta. GABA and malate appear to interact with ALMTs in a complex manner to regulate each other's transport, suggestive of a role for ALMTs in communicating metabolic status.


Impact of the Centers for Disease Control's HIV Preexposure Prophylaxis Guidelines for Men Who Have Sex With Men in the United States.
Preexposure prophylaxis (PrEP) is effective for preventing human immunodeficiency virus (HIV) infection among men who have sex with men (MSM) within trial settings. Population impact will depend on clinical indications for PrEP initiation, coverage levels, and drug adherence. No modeling studies have estimated the impact of clinical practice guidelines for PrEP issued by the Centers for Disease Control and Prevention (CDC). Mathematical models of HIV transmission among MSM were used to estimate the percentage of infections averted (PIA) and the number needed to treat (NNT) under behavioral indications of the CDC's PrEP guidelines. We modeled the contribution of these indications while varying treatment coverage and adherence. At 40% coverage of indicated MSM over the next decade, application of CDC guidelines would avert 1162 infections per 100 000 person-years, 33.0% of expected infections. The predicted NNT for the guidelines would be 25. Increasing coverage and adherence jointly raise the PIA, but reductions to the NNT were associated with better adherence only. Implementation of CDC PrEP guidelines would result in strong and sustained reductions in HIV incidence among MSM in the United States. The guidelines strike a good balance between epidemiological impact (PIA) and efficiency (NNT) at plausible scale-up levels. Adherence counseling could maximize public health investment in PrEP by decreasing the NNT.


Characteristics of Neonates with Cardiopulmonary Disease Who Experience Seizures: A Multicenter Study.
To compare key seizure and outcome characteristics between neonates with and without cardiopulmonary disease. The Neonatal Seizure Registry is a multicenter, prospectively acquired cohort of neonates with clinical or electroencephalographic (EEG)-confirmed seizures. Cardiopulmonary disease was defined as congenital heart disease, congenital diaphragmatic hernia, and exposure to extracorporeal membrane oxygenation. We assessed continuous EEG monitoring strategy, seizure characteristics, seizure management, and outcomes for neonates with and without cardiopulmonary disease. We evaluated 83 neonates with cardiopulmonary disease and 271 neonates without cardiopulmonary disease. Neonates with cardiopulmonary disease were more likely to have EEG-only seizures (40% vs 21%, P < .001) and experience their first seizure later than those without cardiopulmonary disease (174 vs 21 hours of age, P < .001), but they had similar seizure exposure (many-recurrent electrographic seizures 39% vs 43%, P = .27). Phenobarbital was the primary initial antiseizure medication for both groups (90%), and both groups had similarly high rates of incomplete response to initial antiseizure medication administration (66% vs 68%, P = .75). Neonates with cardiopulmonary disease were discharged from the hospital later (hazard ratio 0.34, 95% CI 0.25-0.45, P < .001), although rates of in-hospital mortality were similar between the groups (hazard ratio 1.13, 95% CI 0.66-1.94, P = .64). Neonates with and without cardiopulmonary disease had a similarly high seizure exposure, but neonates with cardiopulmonary disease were more likely to experience EEG-only seizures and had seizure onset later in the clinical course. Phenobarbital was the most common seizure treatment, but seizures were often refractory to initial antiseizure medication. These data support guidelines recommending continuous EEG in neonates with cardiopulmonary disease and indicate a need for optimized therapeutic strategies.


Diversification in substrate usage by glutathione synthetases from soya bean (Glycine max), wheat (Triticum aestivum) and maize (Zea mays).
Unlike animals which accumulate glutathione (gamma-glutamyl-L-cysteinyl-glycine) alone as their major thiol antioxidant, several crops synthesize alternative forms of glutathione by varying the carboxy residue. The molecular basis of this variation is not well understood, but the substrate specificity of the respective GSs (glutathione synthetases) has been implicated. To investigate their substrate tolerance, five GS-like cDNAs have been cloned from plants that can accumulate alternative forms of glutathione, notably soya bean [hGSH (homoglutathione or gamma-glutamyl-L-cysteinyl-beta-alanine)], wheat (hydroxymethylglutathione or gamma-glutamyl-L-cysteinyl-serine) and maize (gamma-Glu-Cys-Glu). The respective recombinant GSs were then assayed for the incorporation of differing C-termini into gamma-Glu-Cys. The soya bean enzyme primarily incorporated beta-alanine to form hGSH, whereas the GS enzymes from cereals preferentially catalysed the formation of glutathione. However, when assayed with other substrates, several GSs and one wheat enzyme in particular were able to synthesize a diverse range of glutathione variants by incorporating unusual C-terminal moieties including D-serine, non-natural amino acids and alpha-amino alcohols. Our results suggest that plant GSs are capable of producing a diverse range of glutathione homologues depending on the availability of the acyl acceptor.


Quantifying the physical activity energy expenditure of commuters using a combination of global positioning system and combined heart rate and movement sensors.
Active commuting may help to increase adults' physical activity levels. However, estimates of its energy cost are derived from a small number of studies which are laboratory-based or use self-reported measures. Adults working in Cambridge (UK) recruited through a predominantly workplace-based strategy wore combined heart rate and movement sensors and global positioning system (GPS) devices for one week, and completed synchronous day-by-day travel diaries in 2010 and 2011. Commuting journeys were delineated using GPS data, and metabolic intensity (standard metabolic equivalents; MET) was derived and compared between journey types using mixed-effects linear regression. 182 commuting journeys were included in the analysis. Median intensity was 1.28 MET for car journeys; 1.67 MET for bus journeys; 4.61 MET for walking journeys; 6.44 MET for cycling journeys; 1.78 MET for journeys made by car in combination with walking; and 2.21 MET for journeys made by car in combination with cycling. The value for journeys made solely by car was significantly lower than those for all other journey types (p<0.04). On average, 20% of the duration of journeys incorporating any active travel (equating to 8 min) was spent in moderate-to-vigorous physical activity (MVPA). We have demonstrated how GPS and activity data from a free-living sample can be used simultaneously to provide objective estimates of commuting energy expenditure. On average, incorporating walking or cycling into longer journeys provided over half the weekly recommended activity levels from the commute alone. This may be an efficient way of achieving physical activity guidelines and improving population health.


Rv1818c-encoded PE_PGRS protein of Mycobacterium tuberculosis is surface exposed and influences bacterial cell structure.
Identification of the novel PE multigene family was an unexpected finding of the genomic sequencing of Mycobacterium tuberculosis. Presently, the biological role of the PE and PE_PGRS proteins encoded by this unique family of mycobacterial genes remains unknown. In this report, a representative PE_PGRS gene (Rv1818c/PE_PGRS33) was selected to investigate the role of these proteins. Cell fractionation studies and fluorescence analysis of recombinant strains of Mycobacterium smegmatis and M. tuberculosis expressing green fluorescent protein (GFP)-tagged proteins indicated that the Rv1818c gene product localized in the mycobacterial cell wall, mostly at the bacterial cell poles, where it is exposed to the extracellular milieu. Further analysis of this PE_PGRS protein showed that the PE domain is necessary for subcellular localization. In addition, the PGRS domain, but not PE, affects bacterial shape and colony morphology when Rv1818c is overexpressed in M. smegmatis and M. tuberculosis. Taken together, the results indicate that PE_PGRS and PE proteins can be associated with the mycobacterial cell wall and influence cellular structure as well as the formation of mycobacterial colonies. Regulated expression of PE genes could have implications for the survival and pathogenesis of mycobacteria within the human host and in other environmental niches.


An Extracellular Redox Signal Triggers Calcium Release and Impacts the Asexual Development of Toxoplasma gondii.
The ability of an organism to sense and respond to environmental redox fluctuations relies on a signaling network that is incompletely understood in apicomplexan parasites such as Toxoplasma gondii. The impact of changes in redox upon the development of this intracellular parasite is not known. Here, we provide a revised collection of 58 genes containing domains related to canonical antioxidant function, with their encoded proteins widely dispersed throughout different cellular compartments. We demonstrate that addition of exogenous H2O2 to human fibroblasts infected with T. gondii triggers a Ca2+ flux in the cytosol of intracellular parasites that can induce egress. In line with existing models, egress triggered by exogenous H2O2 is reliant upon both Calcium-Dependent Protein Kinase 3 and diacylglycerol kinases. Finally, we show that the overexpression a glutaredoxin-roGFP2 redox sensor fusion protein in the parasitophorous vacuole severely impacts parasite replication. These data highlight the rich redox network that exists in T. gondii, evidencing a link between extracellular redox and intracellular Ca2+ signaling that can culminate in parasite egress. Our findings also indicate that the redox potential of the intracellular environment contributes to normal parasite growth. Combined, our findings highlight the important role of redox as an unexplored regulator of parasite biology.


Musical emotions in congenital amusia: Impaired recognition, but preserved emotional intensity.
To further our understanding of the role of perceptual processes in musical emotions, we investigated individuals with congenital amusia, a neurodevelopmental disorder that alters pitch processing. Amusic and matched control participants were studied for emotion recognition and emotion intensity ratings of both musical excerpts and faces. Emotion recognition was found to be impaired in amusic participants relative to controls for the musical stimuli only. This impairment suggests that perceptual deficits in music processing reduce amusics' access to a verbal and conscious representation of musical emotions. Nevertheless, amusics' performance for emotion recognition was above chance level, and multidimensional scaling (MDS) analyses revealed that their categorization of musical pieces was based on similar representation spaces of emotions as for control participants. The emotion intensity ratings, nonverbal and possibly more implicit than the categorization task, seemed to be intact in amusic participants. These findings reveal that pitch deficits can hinder the recognition of emotions conveyed by musical pieces, while also highlighting the (at least partial) dissociation between emotion recognition and emotion intensity evaluation. Our study thus sheds light on the complex interactions between perceptual and emotional networks in the brain, by showing that impaired central auditory processing partially alters musical emotion processing. (PsycINFO Database Record (c) 2018 APA, all rights reserved).


The CD53 and CEACAM-1 genes are genetic targets for early B cell factor.
Early B cell factor (EBF)-1 is a transcription factor known to be of critical importance for early B lymphocyte development. EBF-1 has been shown to directly interact with and regulate expression of a set of genes involved in the functional formation of the pre-B cell receptor, but the dramatic phenotype observed in the EBF-1-deficient mice suggests that several additional genes are activated by this protein. In order to identify additional target genes for EBF-1, we transduced a hematopoietic progenitor cell line, BaF/3, with an EBF-1-encoding retrovirus and investigated the induced gene expression pattern by micro-arrays. This analysis suggested that among others, the CD53 and the carcinoembryonic antigen-related cell adhesion molecule (CEACAM)-1 genes both were induced by ectopic expression of EBF-1. Identification of the 5' end of the cDNA enabled the identification of promoter elements with functional binding sites for EBF-1 and ability to respond to EBF-1 expression in transient transfection assays. These data suggest that CD53 and CEACAM-1 are direct genetic targets for EBF-1, providing additional information concerning the activity of this crucial transcription factor in hematopoiesis.


"Seeing" artificial hydration and nutrition through an ethical lens.
Nurses may be asked by conscious, rational patients to participate in the withdrawal of their life-sustaining nutritional support. The case study presented in this column gives clinicians a framework to handle these situations when they arise, using an ethical framework as their guide.


PROSPECT guideline for elective caesarean section: updated systematic review and procedure-specific postoperative pain management recommendations.
Caesarean section is associated with moderate-to-severe postoperative pain, which can influence postoperative recovery and patient satisfaction as well as breastfeeding success and mother-child bonding. The aim of this systematic review was to update the available literature and develop recommendations for optimal pain management after elective caesarean section under neuraxial anaesthesia. A systematic review utilising procedure-specific postoperative pain management (PROSPECT) methodology was undertaken. Randomised controlled trials published in the English language between 1 May 2014 and 22 October 2020 evaluating the effects of analgesic, anaesthetic and surgical interventions were retrieved from MEDLINE, Embase and Cochrane databases. Studies evaluating pain management for emergency or unplanned operative deliveries or caesarean section performed under general anaesthesia were excluded. A total of 145 studies met the inclusion criteria. For patients undergoing elective caesarean section performed under neuraxial anaesthesia, recommendations include intrathecal morphine 50-100 µg or diamorphine 300 µg administered pre-operatively; paracetamol; non-steroidal anti-inflammatory drugs; and intravenous dexamethasone administered after delivery. If intrathecal opioid was not administered, single-injection local anaesthetic wound infiltration; continuous wound local anaesthetic infusion; and/or fascial plane blocks such as transversus abdominis plane or quadratus lumborum blocks are recommended. The postoperative regimen should include regular paracetamol and non-steroidal anti-inflammatory drugs with opioids used for rescue. The surgical technique should include a Joel-Cohen incision; non-closure of the peritoneum; and abdominal binders. Transcutaneous electrical nerve stimulation could be used as analgesic adjunct. Some of the interventions, although effective, carry risks, and consequentially were omitted from the recommendations. Some interventions were not recommended due to insufficient, inconsistent or lack of evidence. Of note, these recommendations may not be applicable to unplanned deliveries or caesarean section performed under general anaesthesia.


Occupational repetitive strain injuries in Hong Kong.
To review currently available evidence on the epidemiology and management of occupational repetitive strain injuries with particular reference to Hong Kong. Medline, PubMed and Cochrane Library searches of local and internationally published English journals from 1990 to 2007 regarding repetitive strain injuries. All articles involving occupational repetitive strain injuries in Hong Kong were included in this review. There were 16 articles contributing data on the impact of repetitive strain injuries both in Hong Kong and around the world. There were seven articles dealing with the problem of computer station set-ups and methods for improving the workstation environments. Currently there were significant data on the impact of repetitive strain injuries in Hong Kong. The data took the form of compensation claims, days away from work, and cost of medical consultations. Other articles described proper workplace adjustments to help prevent repetitive strain injuries. However, there were no figures in the current literature showing the impact of these adjustments in reducing the incidence of repetitive strain injuries. More research could help to delineate the relationship between different types of interventions and occupational repetitive strain injuries.


Mechanisms of structure generation during plastic compression of nanofibrillar collagen hydrogel scaffolds: towards engineering of collagen.
Operator control of cell/matrix density of plastically compressed collagen hydrogel scaffolds critically depends on reproducibly limiting the extent of scaffold compaction, as fluid expulsion. A functional model of the compression process is presented, based on the idea that the main fluid-leaving surface (FLS) behaves as an ultrafiltration membrane, allowing fluid (water) out but retaining collagen fibrils to form a cake. We hypothesize that accumulation of collagen at the FLS produces anisotropic structuring but also increases FLS hydraulic resistance (R(FLS) ), in turn limiting the flux. Our findings show that while compressive load is the primary determinant of flux at the beginning of compression (load-dependent phase), increasing FLS collagen density (measured by X-ray attenuation) and increasing R(FLS) become the key determinants of flux as the process proceeds (flow-dependent phase). The model integrates these two phases and can closely predict fluid loss over time for a range of compressive loads. This model provides a useful tool for engineering cell and matrix density to tissue-specific levels, as well as generating localized 3D nano micro-scale structures and zonal heterogeneity within scaffolds. Such structure generation is important for complex tissue engineering and forms the basis for process automation and up-scaling.


Changes in the dose of inhaled steroid affect exhaled nitric oxide levels in asthmatic patients.
An increased concentration of nitric oxide (NO) in the exhaled air of asthmatic patients may reflect inflammation of the airways, and exhaled NO may, therefore, be useful in monitoring asthma control and the optimal use of anti-inflammatory treatment. We have studied the effect of reducing and then increasing the dose of inhaled steroid on exhaled NO, lung function and symptoms in 14 asthmatic patients treated with twice daily budesonide. Baseline measurements were made at the end of a 2 week run-in period, 2 weeks after the daily dose of budesonide was reduced by 200 micrograms daily, and 2 weeks after the dose was then increased by 200 micrograms daily. Exhaled NO increased significantly compared with baseline after the dose was reduced by 200 micrograms daily (from 122 +/- 13 to 246 +/- 52 ppb); whereas, there was no significant decrease in spirometry or change in peak flow variability. There was also a significant increase in symptoms at night, but no change during the day or in the number of rescue doses of inhaled beta 2-agonist. The level of exhaled NO decreased when the dose of inhaled steroids was increased, and this was associated with a reduction in diurnal variability of peak expiratory flow, and in nocturnal symptoms. Our study suggests that exhaled nitric oxide may be a useful means of monitoring control of asthma. Further longitudinal studies in patients of differing asthma severity are now indicated.


Excited state non-adiabatic dynamics of the smallest polyene, trans 1,3-butadiene. II. Ab initio multiple spawning simulations.
The excited state non-adiabatic dynamics of the smallest polyene, trans 1,3-butadiene (BD), has long been the subject of controversy due to its strong coupling, ultrafast time scales and the difficulties that theory faces in describing the relevant electronic states in a balanced fashion. Here we apply Ab Initio Multiple Spawning (AIMS) using state-averaged complete active space multistate second order perturbation theory [SA-3-CAS(4/4)-MSPT2] which describes both static and dynamic electron correlation effects, providing a balanced description of both the initially prepared bright 11Bu (ππ*) state and non-adiabatically coupled dark 21Ag state of BD. Importantly, AIMS allows for on-the-fly calculations of experimental observables. We validate our approach by directly simulating the time resolved photoelectron-photoion coincidence spectroscopy results presented in Paper I [A. E. Boguslavskiy et al., J. Chem. Phys. 148, 164302 (2018)], demonstrating excellent agreement with experiment. Our simulations reveal that the initial excitation to the 11Bu state rapidly evolves via wavepacket dynamics that follow both bright- and dark-state pathways as well as mixtures of these. In order to test the sensitivity of the AIMS results to the relative ordering of states, we considered two hypothetical scenarios biased toward either the bright 1Bu or the dark 21Ag state. In contrast with AIMS/SA-3-CAS(4/4)-MSPT2 simulations, neither of these scenarios yields favorable agreement with experiment. Thus, we conclude that the excited state non-adiabatic dynamics in BD involves both of these ultrafast pathways.


Global Vascular Guidelines on the Management of Chronic Limb-Threatening Ischemia.
Chronic limb-threatening ischemia (CLTI) is associated with mortality, amputation, and impaired quality of life. These Global Vascular Guidelines (GVG) are focused on definition, evaluation, and management of CLTI with the goals of improving evidence-based care and highlighting critical research needs. The term CLTI is preferred over critical limb ischemia, as the latter implies threshold values of impaired perfusion rather than a continuum. CLTI is a clinical syndrome defined by the presence of peripheral artery disease (PAD) in combination with rest pain, gangrene, or a lower limb ulceration >2 weeks duration. Venous, traumatic, embolic, and nonatherosclerotic etiologies are excluded. All patients with suspected CLTI should be referred urgently to a vascular specialist. Accurately staging the severity of limb threat is fundamental, and the Society for Vascular Surgery Threatened Limb Classification system, based on grading of Wounds, Ischemia, and foot Infection (WIfI) is endorsed. Objective hemodynamic testing, including toe pressures as the preferred measure, is required to assess CLTI. Evidence-based revascularization (EBR) hinges on three independent axes: Patient risk, Limb severity, and ANatomic complexity (PLAN). Average-risk and high-risk patients are defined by estimated procedural and 2-year all-cause mortality. The GVG proposes a new Global Anatomic Staging System (GLASS), which involves defining a preferred target artery path (TAP) and then estimating limb-based patency (LBP), resulting in three stages of complexity for intervention. The optimal revascularization strategy is also influenced by the availability of autogenous vein for open bypass surgery. Recommendations for EBR are based on best available data, pending level 1 evidence from ongoing trials. Vein bypass may be preferred for average-risk patients with advanced limb threat and high complexity disease, while those with less complex anatomy, intermediate severity limb threat, or high patient risk may be favored for endovascular intervention. All patients with CLTI should be afforded best medical therapy including the use of antithrombotic, lipid-lowering, antihypertensive, and glycemic control agents, as well as counseling on smoking cessation, diet, exercise, and preventive foot care. Following EBR, long-term limb surveillance is advised. The effectiveness of nonrevascularization therapies (eg, spinal stimulation, pneumatic compression, prostanoids, and hyperbaric oxygen) has not been established. Regenerative medicine approaches (eg, cell, gene therapies) for CLTI should be restricted to rigorously conducted randomizsed clinical trials. The GVG promotes standardization of study designs and end points for clinical trials in CLTI. The importance of multidisciplinary teams and centers of excellence for amputation prevention is stressed as a key health system initiative.


The Enigmatic P450 Decarboxylase OleT Is Capable of, but Evolved To Frustrate, Oxygen Rebound Chemistry.
OleT is a cytochrome P450 enzyme that catalyzes the removal of carbon dioxide from variable chain length fatty acids to form 1-alkenes. In this work, we examine the binding and metabolic profile of OleT with shorter chain length (n ≤ 12) fatty acids that can form liquid transportation fuels. Transient kinetics and product analyses confirm that OleT capably activates hydrogen peroxide with shorter substrates to form the high-valent intermediate Compound I and largely performs C-C bond scission. However, the enzyme also produces fatty alcohol side products using the high-valent iron oxo chemistry commonly associated with insertion of oxygen into hydrocarbons. When presented with a short chain fatty acid that can initiate the formation of Compound I, OleT oxidizes the diagnostic probe molecules norcarane and methylcyclopropane in a manner that is reminiscent of reactions of many CYP hydroxylases with radical clock substrates. These data are consistent with a decarboxylation mechanism in which Compound I abstracts a substrate hydrogen atom in the initial step. Positioning of the incipient substrate radical is a crucial element in controlling the efficiency of activated OH rebound.


Male and female child murderers: an empirical analysis of U.S. arrest data.
Recent U.S. cases of murders by children below age 11 have captured national headlines. A review of the literature reveals that little is known about this population of juvenile homicide offenders (JHOs). Most studies on juvenile murderers have used small clinical samples, focused on adolescents, and concentrated on male offenders. Studies that have used Supplementary Homicide Report (SHR) data have found significant gender differences among juveniles below 18 years arrested for murder. This study investigated gender differences among 226 juvenile murderers ages 6 through 10 involved in single-victim incidents using bivariate and multivariate statistical techniques. Consistent with previous research, bivariate analyses revealed gender differences with respect to the type of weapon used, age of the victim, relationship to the victim, and circumstances of the crime. Logistic regression analysis identified female JHOs as more likely to use a knife, kill a family member, and kill a victim below age 5, when compared with male JHOs. From these findings, profiles of young male and female JHOs can be drawn. The article concludes with a discussion of the study's implications for prevention and treatment. The authors recommend that future research in gender differences among young children focus on examining psychological, neurological, and sociological variables not included in the SHR data set.


A new high resolution color flow system using an eigendecomposition-based adaptive filter for clutter rejection.
We present a new signal processing strategy for high frequency color flow mapping in moving tissue environments. A new application of an eigendecomposition-based clutter rejection filter is presented with modifications to deal with high blood-to-clutter ratios (BCR). Additionally, a new method for correcting blood velocity estimates with an estimated tissue motion profile is detailed. The performance of the clutter filter and velocity estimation strategies is quantified using a new swept-scan signal model. In vivo color flow images are presented to illustrate the potential of the system for mapping blood flow in the microcirculation with external tissue motion.


Epigenetic regulation of active Chinese herbal components for cancer prevention and treatment: A follow-up review.
Epigenetic modifications include DNA methylation, histone modification, and other patterns. These processes are associated with carcinogenesis and cancer progression. Thus, epigenetic modification-related enzymes, such as DNA methyltransferases (DNMTs), histone methyltransferases (HMTs), histone demethylases (HDMTs), histone acetyltransferases (HATs), and histone deacetylases (HDACs), as well as some related proteins, including methyl-CpG binding proteins (MBPs) and DNMT1-associated protein (DMAP 1), are considered as potential targets for cancer prevention and therapy. Numerous natural compounds, mainly derived from Chinese herbs and chemically ranging from polyphenols and flavonoids to mineral salts, inhibit the growth and development of various cancers by targeting multiple genetic and epigenetic alterations. This review summarizes the epigenetic mechanisms by which active compounds from Chinese herbs exert their anti-cancer effect. A subset of these compounds, such as curcumin and resveratrol, affect multiple epigenetic processes, including DNMT inhibition, HDAC inactivation, MBP suppression, HAT activation, and microRNA modulation. Other compounds also regulate epigenetic modification processes, but the underlying mechanisms and clear targets remain unknown. Accordingly, further studies are required.


Clinical Profile and Consequences of Atrial Fibrillation in Hypertrophic Cardiomyopathy.
Atrial fibrillation (AF), the most common sustained arrhythmia in hypertrophic cardiomyopathy (HCM), is capable of producing symptoms that impact quality of life and is associated with risk for embolic stroke. However, the influence of AF on clinical course and outcome in HCM remains incompletely resolved. Records of 1558 consecutive patients followed at the Tufts Medical Center Hypertrophic Cardiomyopathy Institute for 4.8±3.4 years (from 2004 to 2014) were accessed. Of the 1558 patients with HCM, 304 (20%) had episodes of AF, of which 226 (74%) were confined to symptomatic paroxysmal AF (average, 5±5; range, 1 to >20), whereas 78 (26%) developed permanent AF, preceded by 7±6 paroxysmal AF episodes. At last evaluation, 277 patients (91%) are alive at 62±13 years of age, including 89% in New York Heart Association class I or II. No difference was found in outcome measures for patients with AF and age- and sex-matched patients with HCM without AF. Four percent of patients with AF died of HCM-related causes (n=11), with annual mortality 0.7%; mortality directly attributable to AF (thromboembolism without prophylactic anticoagulation) was 0.1% per year (n=2 patients). Patients were treated with antiarrhythmic drugs (most commonly amiodarone [n=103] or sotalol [n=78]) and AF catheter ablation (n=49) or the Maze procedure at surgical myectomy (n=72). Freedom from AF recurrence at 1 year was 44% for ablation patients and 75% with the Maze procedure (P<0.001). Embolic events were less common with anticoagulation prophylaxis (4/233, 2%) than without (9/66, 14%) (P<0.001). Transient symptomatic episodes of AF, often responsible for impaired quality of life, are unpredictable in frequency and timing, but amenable to effective contemporary treatments, and infrequently progress to permanent AF. AF is not a major contributor to heart failure morbidity or a cause of arrhythmic sudden death; when treated, it is associated with low disease-related mortality, no different than for patients without AF. AF is an uncommon primary cause of death in HCM virtually limited to embolic stroke, supporting a low threshold for initiating anticoagulation therapy.


Fronto-limbic effective connectivity as possible predictor of antidepressant response to SSRI administration.
The timely selection of the optimal treatment for depressed patients is critical to improve remission rates. The detection of pre-treatment variables able to predict differential treatment response may provide novel approaches for treatment selection. Selective serotonin reuptake inhibitors (SSRIs) modulate the fronto-limbic functional response and connectivity, an effect preceding the overt clinical antidepressant effects. Here we investigated whether the cortico-limbic connectivity associated with emotional bias measured before SSRI administration predicts the efficacy of antidepressant treatment in MDD patients. fMRI and Dynamic Causal Modeling (DCM) were combined to study if effective connectivity might differentiate healthy controls (HC) and patients affected by major depression who later responded (RMDD, n=21), or failed to respond (nRMDD, n=12), to 6 weeks of escitalopram administration. Sixteen DCMs exploring connectivity between anterior cingulate cortex (ACC), ventrolateral prefrontal cortex (VLPFC), Amygdala (Amy), and fusiform gyrus (FG) were constructed. Analyses revealed that nRMDD had reduced endogenous connectivity from Amy to VLPFC and to ACC, with an increased connectivity and modulation of the ACC to Amy connectivity when processing of fearful emotional stimuli compared to HC. RMDD and HC did not significantly differ among themselves. Pre-treatment effective connectivity in fronto-limbic circuitry could be an important factor affecting antidepressant response, and highlight the mechanisms which may be involved in recovery from depression. These results suggest that fronto-limbic connectivity might provide a neural biomarker to predict the clinical outcome to SSRIs administration in major depression.


Insulin-like growth factor levels in cord blood, birth weight and breast cancer risk.
Breast cancer incidence and birth weight are higher among Caucasian than Asian women, and birth size has been positively associated with breast cancer risk. Pregnancy hormone levels, however, have been generally lower in Caucasian than Asian women. We studied components of the insulin-like growth factor (IGF) system in cord blood from 92 singleton babies born in Boston, USA, and 110 born in Shanghai, China, in 1994-1995. Cord blood IGF-1 was significantly higher among Caucasian compared with Chinese babies (P<10(-6)). The opposite was noted for IGF-2 (P approximately 10(-4)). IGF-1 was significantly positively associated with birth weight and birth length in Boston, but not Shanghai. In contrast, stronger positive, though statistically non-significant, associations of IGF-2 with birth size were only evident in Shanghai. The associations of birth weight and birth length were positive and significant in taller women (for IGF-1 in Boston P approximately 0.003 and 0.03, respectively; for IGF-2 in Shanghai P approximately 0.05 and approximately 0.04, respectively), among whom maternal anthropometry does not exercise strong constraints in foetal growth. The documentation of higher cord blood levels of IGF-1, a principal growth hormone that does not cross the placenta, among Caucasian than in Asian newborns is concordant with breast cancer incidence in these populations.


Burkholderia pseudomallei sequencing identifies genomic clades with distinct recombination, accessory, and epigenetic profiles.
Burkholderia pseudomallei (Bp) is the causative agent of the infectious disease melioidosis. To investigate population diversity, recombination, and horizontal gene transfer in closely related Bp isolates, we performed whole-genome sequencing (WGS) on 106 clinical, animal, and environmental strains from a restricted Asian locale. Whole-genome phylogenies resolved multiple genomic clades of Bp, largely congruent with multilocus sequence typing (MLST). We discovered widespread recombination in the Bp core genome, involving hundreds of regions associated with multiple haplotypes. Highly recombinant regions exhibited functional enrichments that may contribute to virulence. We observed clade-specific patterns of recombination and accessory gene exchange, and provide evidence that this is likely due to ongoing recombination between clade members. Reciprocally, interclade exchanges were rarely observed, suggesting mechanisms restricting gene flow between clades. Interrogation of accessory elements revealed that each clade harbored a distinct complement of restriction-modification (RM) systems, predicted to cause clade-specific patterns of DNA methylation. Using methylome sequencing, we confirmed that representative strains from separate clades indeed exhibit distinct methylation profiles. Finally, using an E. coli system, we demonstrate that Bp RM systems can inhibit uptake of non-self DNA. Our data suggest that RM systems borne on mobile elements, besides preventing foreign DNA invasion, may also contribute to limiting exchanges of genetic material between individuals of the same species. Genomic clades may thus represent functional units of genetic isolation in Bp, modulating intraspecies genetic diversity.


Glucitol-core containing gallotannins inhibit the formation of advanced glycation end-products mediated by their antioxidant potential.
Glucitol-core containing gallotannins (GCGs) are polyphenols containing galloyl groups attached to a 1,5-anhydro-d-glucitol core, which is uncommon among naturally occurring plant gallotannins. GCGs have only been isolated from maple (Acer) species, including the red maple (Acer rubrum), a medicinal plant which along with the sugar maple (Acer saccharum), are the major sources of the natural sweetener, maple syrup. GCGs are reported to show antioxidant, α-glucosidase inhibitory, and antidiabetic effects, but their antiglycating potential is unknown. Herein, the inhibitory effects of five GCGs (containing 1-4 galloyls) on the formation of advanced glycation end-products (AGEs) were evaluated by MALDI-TOF mass spectroscopy, and BSA-fructose, and G.K. peptide-ribose assays. The GCGs showed superior activities compared to the synthetic antiglycating agent, aminoguanidine (IC50 15.8-151.3 vs. >300 μM) at the early, middle, and late stages of glycation. Circular dichroism data revealed that the GCGs were able to protect the secondary structure of BSA protein from glycation. The GCGs did not inhibit AGE formation by the trapping of reactive carbonyl species, namely, methylglyoxal, but showed free radical scavenging activities in the DPPH assay. The free radical quenching properties of the GCGs were further confirmed by electron paramagnetic resonance spectroscopy using ginnalin A (contains 2 galloyls) as a representative GCG. In addition, this GCG chelated ferrous iron, an oxidative catalyst of AGE formation, supported a potential antioxidant mechanism of antiglycating activity for these polyphenols. Therefore, GCGs should be further investigated for their antidiabetic potential given their antioxidant, α-glucosidase inhibitory, and antiglycating properties.


Relationship between reproductive success and male plasma vitellogenin concentrations in cunner, Tautogolabrus adspersus.
The gene for vitellogenin, an egg yolk protein precursor, is usually silent in male fish but can be induced by estrogen exposure. For this reason, vitellogenin production in male fish has become a widely used indicator of exposure to exogenous estrogens or estrogen mimics in the aquatic environment. The utility of this indicator to predict impacts on fish reproductive success is unclear because information on the relationship between male plasma vitellogenin and reproductive end points in male and female fish is limited. In the research reported in this article, we investigated whether the presence of male plasma vitellogenin is a reliable indicator of decreased reproductive success in mature fish. Adult and sexually mature male and female cunner (Tautogolabrus adspersus) were exposed to 17ss-estradiol, ethynylestradiol, or estrone, three steroidal estrogens that elicit the vitellogenic response. Data were gathered and pooled on egg production, egg viability, egg fertility, sperm motility, and male plasma vitellogenin concentrations. All males, including two with plasma vitellogenin levels exceeding 300 mg/mL, produced motile sperm. Neither percent fertile eggs nor percent viable eggs produced by reproductively active fish demonstrated a significant correlation with male plasma vitellogenin concentrations. Male gonadosomatic index and average daily egg production by females showed significant, but weak, negative correlation with male plasma vitellogenin concentrations. Results suggest that male plasma vitellogenin expression is not a reliable indicator of male reproductive dysfunction in adult cunner exposed to estrogens for 2-8 weeks during their reproductive season, at least in relation to capacity to produce motile sperm or fertilize eggs. Male plasma vitellogenin expression may serve as an indicator of reduced female reproductive function caused by estrogen exposure.


Altered expression of matrix metalloproteinases and tight junction proteins in rats following PEMF-induced BBB permeability change.
To investigate the expression of occludin, ZO-1, MMP-2, and MMP-9 in cerebral microvasculature following Pulse Electromagnetic Field (PEMF) induced BBB permeability change. Sprague-Dawley rats were randomized into PEMF and sham exposed groups (n = 8). After exposure to PEMF at 0.5, 1, 3, 6, and 12 h, BBB permeability was measured by Evans-Blue extravasation. The expression of occludin, ZO-1, MMP-2, and MMP-9 were detected by real-time quantitative reverse transcriptase PCR and western blotting. MMP-2 and MMP-9 activity were detected by EnzChek gelatinase assay. Compared with the sham group, PEMF exposure led to increased permeability of the BBB to EB, which was prolonged after exposure. BBB permeability became progressively more severe, and recovered at 6 h. The gene and protein expression of occludin and ZO-1 were significantly decreased, while MMP-2 and MMP-9 expression were significantly increased after exposure to PEMF. All levels of expression recovered 12 h following PEMF. Changes to BBB permeability were related to the alteration expression of tight junction proteins and matrix metalloproteinase after exposure to PEMF.


Benefits and costs of screening Ashkenazi Jewish women for BRCA1 and BRCA2.
To determine the survival benefit and cost-effectiveness of screening Ashkenazi Jewish women for three specific BRCA1/2 gene mutations. We used a Markov model and Monte Carlo analysis to estimate the survival benefit and cost-effectiveness of screening for three specific mutations in a population in which their prevalence is 2.5% and the associated cancer risks are 56% for breast cancer and 16% for ovarian cancer. We assumed that the sensitivity and specificity of the test were 98% and 99%, respectively, that bilateral prophylactic oophorectomy would reduce ovarian cancer risk by 45%, and that bilateral prophylactic mastectomy would reduce breast cancer risk by 90%. We used Medicare payment data for treatment costs and Surveillance, Epidemiology, and End Results data for cancer survival. Our model suggests that genetic screening of this population could prolong average nondiscounted survival by 38 days (95% probability interval, 22 to 57 days) for combined surgery, 33 days (95% probability interval, 18 to 43 days) for mastectomy, 11 days (95% probability interval, 4 to 25 days) for oophorectomy, and 6 days (95% probability interval, 3 to 8 days) for surveillance. The respective cost-effectiveness ratios per life-year saved, with a discount rate of 3%, are $20,717, $29,970, $72,780, and $134,273. In this Ashkenazi Jewish population, with a high prevalence of BRCA1/2 mutations, genetic screening may significantly increase average survival and, depending on costs and screening/treatment strategies, may be cost-effective by the standards of accepted cancer screening tests. According to our model, screening is cost-effective only if all women who test positive undergo prophylactic surgery. These estimates require confirmation through prospective observational studies and clinical trials.


A Scaffold Merging Approach to Hsp90 C-terminal Inhibition: Synthesis and Evaluation of a Chimeric Library.
Inhibition of the Hsp90 C-terminus is an attractive therapeutic paradigm for the treatment of cancer, however the developmental space of C-terminal inhibitors is limited. It was hypothesized that the combination of two previously identified scaffolds into a single structure could provide a platform for which to probe the three-dimensional space within the Hsp90 C-terminal binding pocket. The resulting chimeric compounds displayed anti-proliferative activity at low micromolar concentrations and manifested inhibitory activity in an Hsp90-dependent rematuration assay. Initial structure-activity relationships suggest that this new scaffold binds Hsp90 in a conformation different from that of the parent compounds, and consequently, provides a new opportunity to develop more efficacious inhibitors of the Hsp90 C-terminal binding pocket.


General road detection from a single image.
Given a single image of an arbitrary road, that may not be well-paved, or have clearly delineated edges, or some a priori known color or texture distribution, is it possible for a computer to find this road? This paper addresses this question by decomposing the road detection process into two steps: the estimation of the vanishing point associated with the main (straight) part of the road, followed by the segmentation of the corresponding road area based upon the detected vanishing point. The main technical contributions of the proposed approach are a novel adaptive soft voting scheme based upon a local voting region using high-confidence voters, whose texture orientations are computed using Gabor filters, and a new vanishing-point-constrained edge detection technique for detecting road boundaries. The proposed method has been implemented, and experiments with 1003 general road images demonstrate that it is effective at detecting road regions in challenging conditions.


Highly electrocatalytic activity of RuO₂ nanocrystals for triiodide reduction in dye-sensitized solar cells.
Dye-sensitized solar cells (DSCs) are promising alternatives to conventional silicon devices because of their simple fabrication procedure, low cost, and high efficiency. Platinum is generally used as a superior counter electrode (CE) material, but the disadvantages such as high cost and low abundance greatly restrict the large-scale application of DSCs. An efficient and sustainable way to overcome the limited supply of Pt is the development of high-efficiency Pt-free CE materials, which should possess both high electrical conductivity and superior electrocatalytic activity simultaneously. Herein, for the first time, a two-step strategy to synthesize ruthenium dioxide (RuO₂) nanocrystals is reported, and it is shown that RuO₂ catalysts exhibit promising electrocatalytic activity towards triiodide reduction, which results in comparable energy conversion efficiency to that of conventional Pt CEs. More importantly, by virtue of first-principles calculations, the catalytic mechanism of electrocatalysis for triiodide reduction on various CEs is investigated systematically and it is found that the electrochemical triiodide reduction reaction on RuO₂ catalyst surfaces can be enhanced significantly, owing to the ideal combination of good electrocatalytic activity and high electrical conductivity.


Effects of teriparatide and bisphosphonate on spinal fusion procedure: A systematic review and network meta-analysis.
Giving patients anti-osteoporotic agents peri-operatively is a well-accepted strategy to increase fusion rate and prevent complications. The purpose of this study was to investigate effectiveness of teriparatide and bisphosphonate on fusion surgery of thoracic and lumbar spine. We searched EMBASE and PubMed for randomized clinical trials (RCTs) and prospective comparative studies using teriparatide or bisphosphonate in peri-operative spinal fusion surgery. Our synthesized data of fusion rate, Oswestry disability index (ODI), and adverse event in contrast-based network meta-analysis. Pooled results were presented in risk ratio (RR) or mean difference (MD) with 95% confidence interval (CI). Our search hit eight RCTs and three prospective studies with 676 patients receiving spinal surgery. Pooled result showed that teriparatide+Denosumab leads to significantly higher fusion rate than placebo (RR, 2.84; 95% CI: 1.22 to 6.60) and bisphosphonate (RR, 2.59; 95% CI: 1.13 to 5.96). We did not observe significant finding among placebo, teriparatide, and bisphosphonate in the two network models. This is the first network meta-analysis providing an overview of the use of teriparatide and bisphosphonate for spinal fusion surgery. Teriparatide treatments are worth to be consider for spinal fusion surgery.


Influence of motion coupling and nonlinear effects on parametric roll for a floating production storage and offloading platform.
Occurrence and features of parametric roll (PR) on a weather-vaning floating production storage and offloading (FPSO) platform with a turret single-point mooring-line system are examined. The main focus is on the relevance of motions coupling and nonlinear effects on this phenomenon and on more general unstable conditions as well as on the occurrence and severity of water on deck. This work was motivated by recent experiments on an FPSO model without mooring systems highlighting the occurrence of parametric resonance owing to roll-yaw coupling. A three-dimensional numerical hybrid potential-flow seakeeping solver was able to capture this behaviour. The same method, extended to include the mooring lines, is adopted here to investigate the platform behaviour for different incident wavelengths, steepnesses, headings, locations of the turret and pretensions. From the results, sway and yaw tend to destabilize the system, also bringing chaotic features. The sway-roll-yaw coupling widens the existence region of PR resonance and increases PR severity; it also results in a larger amount of shipped water, especially at smaller wavelength-to-ship length ratio and larger steepness. The chaotic features are excited when a sufficiently large yaw amplitude is reached. Consistently, a simplified stability analysis showed the relevance of nonlinear-restoring coefficients, first those connected with the sway-yaw coupling then those associated with the roll-yaw coupling, both destabilizing. From the stability analysis, the system is unstable for all longitudinal locations of the turret and pre-tensions examined, but the instability weakens as the turret is moved forward, and the pre-tension is increased. The use of a suitable dynamic-positioning system can control the horizontal motions, avoiding the instability.


Enhancement of in vitro interleukin-2 production in normal subjects following a single spinal manipulative treatment.
Increasing evidence supports somato-visceral effects of manual therapies. We have previously demonstrated that a single spinal manipulative treatment (SMT) accompanied by audible release has an inhibitory effect on the production of proinflammatory cytokines in asymptomatic subjects. The purpose of this study is to report on SMT-related changes in the production of the immunoregulatory cytokine interleukin 2 (IL-2) and to investigate whether such changes might differ with respect to the treatment approach related to the presence or absence of an audible release (joint cavitation). Of 76 asymptomatic subjects, 29 received SMT with cavitation (SMT-C), 23 were treated with SMT without cavitation (SMT-NC) and 24 comprised the venipuncture control (VC) group. The SMT-C and SMT-NC subjects received a single, similar force high velocity low amplitude manipulation, in the upper thoracic spine. However, in SMT-NC subjects, positioning and line of drive were not conducive to cavitation. Blood and serum samples were obtained before and then at 20 and 120 min post-intervention. The production of IL-2 in peripheral blood mononuclear cell cultures was induced by activation for 48 hr with Staphylococcal protein A (SPA) and, in parallel preparations, with the combination of phorbol ester (TPA) and calcium ionophore. The levels of IL-2 in culture supernatants and serum were assessed by specific immunoassays. Compared with VC and their respective baselines, SPA-induced secretion of IL-2 increased significantly in cultures established from both SMT-C and SMT-NC subjects at 20 min post-intervention. At 2 hr post-treatment, significant elevation of IL-2 synthesis was still apparent in preparations from SMT-treated groups though it became somewhat attenuated in SMT-NC subjects. Conversely, IL-2 synthesis induced by TPA and calcium ionophore was unaltered by either type of SMT and was comparable to that in VC group at all time points. No significant alterations in serum-associated IL-2 levels were observed in any of the study groups. The present study demonstrates that, the in vitro T lymphocyte response to a conventional mitogen (SPA), as measured by IL-2 synthesis, can become enhanced following SMT. Furthermore, within a period of time following the manipulative intervention, this effect may be independent of joint cavitation. Thus the results of this study suggest that, under certain physiological conditions, SMT might influence IL-2-regulated biological responses.


An innovative method to generate a Good Manufacturing Practice-ready regulatory T-cell product from non-mobilized leukapheresis donors.
There is real and sustained interest in preparing T-regulatory cells from leukapheresis collections for cellular therapy through the use of simple, effective and reliable methods conforming to Good Manufacturing Practice (GMP). We describe a GMP-ready isolation procedure for CD25(+) products with the use of a sterile docking device, pigtail sampling, a laminar flow hood and the CliniMACS system that uses leukapheresis collections made by two apheresis machines. We used CD8/CD19 depletion followed by CD25-positive selection. The median number of CD4(+) cells recovered was 72.5 ± 32.6 × 10(6), of which 60.5% ± 17.8% were CD25(+)FOXP3(+) cells. Suppression of autologous CD25(-) cell proliferation by the cryopreserved CD25(+) products was 51.3% ± 4.4%, 49.0% ± 3.7% and 39.0% ± 3.6% at CD25(+):CD25(-) ratios of 1:1, 1:2 and 1:4 (n = 6), respectively, comparable to suppression by fresh CD25(+) products (53% ± 6.2%, 51% ± 3.3% and 39% ± 7.1%). We found Leukapheresis collections by Cobe Spectra contained more lymphocytes and platelets than collections by Spectra Optia apheresis machine (median, 9.2 × 10(9) versus 6.7 × 10(9); P = 0.04) and platelets (median, 610 × 10(9) versus 170 × 10(9); P = 0.04). The frequency of CD4(+)CD25(+)FOXP3(+) was significantly higher in the leukapheresis (4.85%; 95% confidence interval, 1.95% to 5.38%) than in peripheral blood (3.9%; 95% confidence interval, 2.63% to 6.45%) (P = 0.02). The CD8- and CD19-negative depletion step was associated with significant loss of total CD4(+) T cells (P = 0.001). Results suggest that functional CD25(+) products can be isolated with a GMP-ready method, and good recovery can be obtained with the use of an optimized cryopreservation protocol. These data and methods show the potential, possibilities and future work needed to isolate target cell populations in a reproducible, time-efficient and cost-efficient manner for clinical applications.


Network theory in autoimmunity. In vitro suppression of serum anti-DNA antibody binding to DNA by anti-idiotypic antibody in systemic lupus erythematosus.
Regulation of serum anti-DNA antibody in systemic lupus erythematosus (SLE) by an antiidiotypic antibody was evaluated. Various sera from SLE patients in active and inactive states of their disease, as well as sera from normal individuals, were first completely depleted of anti-DNA and of DNA by affinity chromatography. The suppressive capacity of equimolar concentrations of the various depleted sera (blocking sera) on target lupus sera were determined. The target sera were from lupus patients with known DNA-binding capacity. Blocking sera from inactive SLE suppressed the binding of autologous anti-DNA antibody to [(3)H]DNA (n = 19,P < 0.01). Blocking sera from active SLE (n = 19), as well as human serum albumin, did not suppress. Sera from normal donors who had no contact with lupus patients or with lupus sera did not suppress (n = 14, P > 0.5), whereas those from normal donors who had contact with lupus patients or sera did suppress the binding (n = 5,P < 0.02). The anti-anti-DNA antibody suppressive activity in the inactive lupus serum was shown to be localized within the F(ab')(2) portion of immunoglobulin (Ig)G and could not be removed upon adsorption by normal human gammaglobulin. Furthermore, immune complexes could be detected by a Clq binding assay when the inactive lupus blocking sera were incubated with the anti-DNA antibody containing target sera. The specificity of the suppressive serum factor was shown by its inability to block the binding of tetanus toxoid to antitetanus antibody and its ability to block the binding of DNA to F(ab')(2) fragments of active lupus IgG. Regulation of serum anti-DNA antibody levels by anti-antibodies could induce and maintain disease remission in lupus patients and prevent disease expression in normals.


Complement C2 receptor inhibitor trispanning confers an increased ability to resist complement-mediated lysis in Trypanosoma cruzi.
The ability to resist complement differs between the Y and Colombiana Trypanosoma cruzi strains. We found that the Y strain of T. cruzi was more able to resist the classical and lectin pathways of complement activation than the Colombiana strain. The complement C2 receptor inhibitor trispanning gene (CRIT) is highly conserved in both strains. At the protein level, CRIT is expressed only in stationary-phase epimastigotes of the Y but not the Colombiana strain and is expressed in infectious metacyclic trypomastigotes of both strains. Y strain epimastigotes with an overexpressed CRIT gene (pTEX-CRIT) had higher survival in normal human serum (NHS). Overexpression of the Y strain CRIT gene in Colombiana epimastigote forms increased the parasite's resistance to lysis mediated by the classical and lectin pathways but not to lysis mediated by alternative pathways. CRIT involvement on the parasite surface was confirmed by showing that the lytic activity of NHS against epimastigotes could be restored by adding excess C2.


How effective are common medications: a perspective based on meta-analyses of major drugs.
The vastness of clinical data and the progressing specialization of medical knowledge may lead to misinterpretation of medication efficacy. To show a realistic perspective on drug efficacy we present meta-analyses on some of the most commonly used pharmacological interventions. For each pharmacological intervention we present statistical indexes (absolute risk or response difference, percentage response ratio, mean difference, standardized mean difference) that are often used to represent efficacy. We found that some of the medications have relatively low effect sizes with only 11 out of 17 of them showing a minimal clinically important difference. Efficacy was often established based on surrogate outcomes and not the more relevant patient-oriented outcomes. As the interpretation of the efficacy of medication is complex, more training for physicians might be needed to get a more realistic view of drug efficacy. That could help prevent harmful overtreatment and reinforce an evidence-based, but personalized medicine.


Changing patterns of knowledge, reported behaviour and sexually transmitted infections in a South African gold mining community.
In 1998, a major HIV intervention project was started in a mining community in Carletonville, South Africa. This included community-based peer education, condom distribution, syndromic management of sexually transmitted infections (STI), and presumptive STI treatment for sex workers. To investigate changes in sexual behaviour and the prevalence of STI before and 2 years after the start of the HIV prevention programme. Cross-sectional surveys were carried out in 1998 and 2000 among mine workers, sex workers and adults in the community. Demographic and behavioural factors were recorded and participants were tested for syphilis, gonorrhoea and chlamydial infection and, at the start of the intervention, for HIV. In 1998, the prevalence of HIV among men and women in the general population, mine workers, and sex workers, was 20%, 37%, 29% and 69%, respectively. In 2000, syphilis, gonorrhoea and chlamydial infection had increased among mine workers; chlamydial infection had increased among men and women, and syphilis had increased among women. There was evidence of positive behaviour change but this was not substantial or universal. Knowledge of HIV/AIDS and awareness of the epidemic were high but condom use remained low. There was little evidence of significant behaviour change and the prevalence of curable STI increased. The prevention programme had had less impact than expected. Reasons for the reduced impact, and the lessons for future intervention projects are discussed. There is a need for further monitoring of the HIV epidemic especially as its impact increases.


Prostate cancer outcomes among older men: insurance status comparisons results from CaPSURE database.
With growing number of older adults in the United States and complexity of issues related to Medicare and other insurances more research is needed to evaluate an effectiveness of the different insurance types in prevention, screening and treatment of cancer. With prostate cancer being highly prevalent disease in older men, the importance of appropriate treatment and favorable outcomes is imperative. In this study we examine whether prostate cancer outcomes, such as risk category at diagnosis, treatment and survival differ in relationship to insurance status in older patients in CaPSURE. Data were abstracted from CaPSURE, a longitudinal observational database of 13 124 men with prostate cancer. Men were selected for the study if they were older than 65 years old at diagnosis, newly diagnosed between 1995 and 2005 at entry to CaPSURE with localized disease and received radical prostatectomy (RP), external beam radiation (EBRT), brachytherapy (BT), hormonal therapy or expectant management (EM). Insurance status was summarized by eight categories: Medicare only, Medicare+supplement, Medicare+HMO, Medicare+PPO, Medicare+FFS, health maintenance organization (HMO), preferred provider organization (PPO) and Veteran's Administration (VA). A total of 2983 men met the inclusion criteria. Odds ratios (OR) for the likelihood of receiving each type of therapy compared to RP by insurance status and likelihood of presenting with high-risk classification at diagnosis were derived using multinomial logistic regression, adjusting for clinical and demographic characteristics. Difference in survival between insurance groups was evaluated by Cox's multivariate regression. Multivariate analysis demonstrated a strong association between initial treatment and insurance status. Compared to Medicare patients, men in the CaPSURE database treated at HMO, PPO and VA systems were more likely to receive BT than RP (OR, 1.71-1.92) and less likely to receive this treatment if they were in Medicare+FFS and Medicare+PPO (OR, 0.18-0.38). Hormonal treatment demonstrated similar pattern, however OR did not reached statistical significance for HMO and PPO. Use of EM was much more predominant for patients in VA system (OR, 4.74; 95% CI, 1.94-11.55). Use of EBRT was significantly associated with type of insurance. Men with VA, Medicare+FFS and Medicare+PPO insurance were less likely to receive this treatment compared to RP. Survival and clinical risk at diagnosis was associated with insurance status in univariate analysis but this association diminished after adjusting for possible covariates. This study provides important information on relationship between insurance status and several outcomes in patients with prostate cancer. Even after controlling for important clinical and sociodemographic factors we found marked differences in prostate cancer treatment according to type of insurance. Future explorations of associations between health care delivery system, cancer care and outcomes are needed.


Next-Generation Methods for HIV Partner Services: A Systematic Review.
Partner notification is a widely accepted method whose intent is to limit onward HIV transmission. With increasing use of new technologies such as text messaging, e-mail, and social network sites, there is growing interest in using these techniques for "next-generation" HIV partner services (PS). We conducted a systematic review to assess the use and effectiveness of these technologies in HIV PS. Our literature search resulted in 1343 citations, with 7 meeting inclusion criteria. We found programs in 2 domains: (1) Public Health Department usage of new technologies to augment traditional partner notification (n = 3) and (2) patient or provider-led usage of partner notification Web sites (n = 4) The health department-based efforts showed an ability to find new cases in a previously unreachable population but in the limited comparisons to traditional PS had a lower rate of successful contact. Usage data from the partner notification Web sites revealed a high total number of e-notifications sent, with less than 10% of cards sent for HIV. Clear evidence on outcomes and directly traceable utilization for these Web services was lacking. When given a choice, most clients chose to send e-notifications via text versus e-mail. Although successful notification may be lower overall, use of next-generation services provides an avenue to contact those who would previously have been untraceable. Additional research is needed to determine to what extent technology-enhanced PS improves the identification of newly infected persons as well as the initiation of new prevention interventions for HIV-negative clients within high-risk networks.


Evidence for the presence of cholesterol side chain cleavage cytochrome P-450 and adrenodoxin in fresh granulosa cells. Effects of follicle-stimulating hormone and cyclic AMP on cholesterol side chain cleavage cytochrome P-450 synthesis and activity.
The synthesis of cholesterol side chain cleavage cytochrome P-450 (cytochrome P-450scc) and adrenodoxin was studied both in freshly harvested bovine granulosa cells and in granulosa cells maintained in primary monolayer culture. In addition, the action of follicle-stimulating hormone (FSH) and cyclic AMP analogs to stimulate the synthesis of cytochrome P-450scc was investigated in cultured cells. Precursor forms of cytochrome P-450scc and adrenodoxin were immunoisolated from a cell-free translation system directed by RNA prepared from freshly obtained granulosa cells that were not luteinized. Furthermore, the presence of cytochrome P-450scc in lysates of granulosa cells freshly obtained from very small follicles (containing less than 0.1 ml of follicular fluid) and in mitochondria of freshly obtained granulosa cells was demonstrated by using an immunoblotting technique. Continuous treatment of cultured granulosa cells with FSH or with cyclic AMP analogs (dibutyryl cyclic AMP or 8-bromo cyclic AMP) for 72 h increased incorporation of [35S]methionine into immunoprecipitable cytochrome P-450scc. Moreover, FSH, dibutyryl cyclic AMP, and 8-bromo cyclic AMP stimulated pregnenolone production by cultured granulosa cells (2.3-, 4.0-, and 7.5-fold increase over control, respectively), indicative of an increase in cholesterol side chain cleavage activity. The results of this study demonstrate for the first time the presence of two components of the cholesterol side chain cleavage system in freshly obtained granulosa cells, and provide direct evidence for the trophic effect of FSH and its presumed mediator, cyclic AMP, on the synthesis of cytochrome P-450scc in granulosa cells.


Technical note: Data loggers are a valid method for assessing the feeding behavior of dairy cows using the Calan Broadbent Feeding System.
Assessing feeding behavior is important in understanding the effects of nutrition and management on the well-being of dairy cows. Historically, collection of these data from cows fed with a Calan Broadbent Feeding System (American Calan Inc., Northwood, NH) required the labor-intensive practices of direct observation or video review. The objective of this study was to evaluate the agreement between the output of a HOBO change-of-state data logger (Onset Computer Corp., Bourne, MA), mounted to the door shell and latch plate, and video data summarized with continuous sampling. Data (number of feed bin visits per day and feeding time in minutes per day) were recorded with both methods from 26 lactating cows and 10 nonlactating cows for 3 d per cow (n=108). The agreement of the data logger and video methods was evaluated using the REG procedure of SAS to compare the mean response of the methods against the difference between the methods. The maximum allowable difference (MAD) was set at ±3 for bin visits and ±20 min for feeding time. Ranges for feed bin visits (2 to 140 per d) and feeding time (28 to 267 min/d) were established from video data. Using the complete data set, agreement was partially established between the data logger and video methods for feed bin visits, but not established for feeding time. The complete data set generated by the data logger was screened to remove visits of a duration ≤3 s, reflecting a cow unable to enter a feed bin (representing 7% of all data) and ≥5,400 s, reflecting a failure of the data logger to align properly with its corresponding magnetic field (representing <1% of all data). Using the resulting screened data set, agreement was established for feed bin visits and feeding time. For bin visits, 4% of the data was beyond the MAD. For feeding time, 3% of the data was beyond the MAD and 74% of the data was ±1 min. The insignificant P-value, low coefficient of determination, and concentration of the data within the MAD indicate the agreement of the change-of-state data logger and video data. This suggests the usage of a change-of-state data logger to assess the feeding behavior of cows feeding from a Calan Broadbent Feeding System is appropriate. Use of the screening criteria for data analysis is recommended.


Ventral tegmental area orexin 1 receptors promote palatable food intake and oppose postingestive negative feedback.
Hypothalamic orexin neurons project to numerous brain areas, including the ventral tegmental area (VTA), which is involved in motivation and food-seeking behavior. Here we address how exogenously administered orexin-A and endogenous orexin 1 receptor (OX1R) activation in the VTA affects feeding behavior. We hypothesized that orexin-A and OX1R antagonist SB334867 delivered to the VTA, at doses that were subthreshold for effect when injected into the ventricle, would affect intake of palatable foods in multiple test situations. We first used a hedonic feeding model in which satiated rats selectively consume a high-fat diet (HFD). Intra-VTA orexin-A stimulated additional consumption of chow and increased HFD intake in this model. In ad libitum-fed rats given daily 30-min test sessions, intra-VTA orexin-A also increased intake of HFD and 0.1 M sucrose. Further analysis of licking patterns revealed that that VTA orexin-A increased meal size and licking burst size only toward the end of the meal. Consistent with this finding, a subthreshold dose of VTA orexin-A prevented intake suppression induced by gastrointestinal nutrient infusion. Surprisingly, intra-VTA orexin-A had no effect on operant responding for sucrose pellets on a progressive ratio schedule of reinforcement. A role for endogenous VTA OX1R stimulation is supported by our finding that bilateral VTA injection of the selective OX1R antagonist SB334867 suppressed 0.1 M sucrose intake. Together, our data suggest that OX1R activity in the VTA facilitates food intake, potentially by counteracting postingestive negative feedback that would normally suppress feeding later in a meal.


Recurrent hepatitis C virus after transplant and the importance of plasma cells on biopsy.
Hepatitis C virus (HCV) is the leading indication for liver transplantation in the United States. It recurs universally after transplant but the rate of fibrosis and the development of graft failure is variable. Different donor and recipient features have been demonstrated to impact fibrosis. Plasma cell hepatitis, a histologic finding, is one feature associated with poor graft and patient outcomes. The pathogenic mechanism resulting in plasma cell hepatitis is poorly understood, with evidence suggesting a role for both the HCV and the immune system.A recent publication described plasma cell hepatitis in a larger context of immune medicated graft dysfunction in transplant recipients receiving interferon based therapy. This manuscript will highlight the topic of plasma cell hepatitis and provide commentary on the lack of recognition, the data regarding pathophysiologic mechanisms and the potential management options.


Role of the cmcH-ccaR intergenic region and ccaR overexpression in cephamycin C biosynthesis in Streptomyces clavuligerus.
The effect of the CcaR regulatory protein on expression of the cephamycin C gene cluster is studied. Quantitative reverse transcription PCR (qRT-PCR) expression analysis of the cephamycin biosynthesis genes in the ccaR-disrupted strain, S. clavuligerus ccaR::aph, revealed that in the absence of CcaR, the lat and cmcI genes expression was reduced 2,200- and 1,087-fold compared with the wild type. Expression of pcbAB-pcbC-cefD-cefE-cmcJ-cmcH and blp was 225- to 359-fold lower, while expression of pcbR-pbpA-bla and orf10 was only slightly affected if at all, indicating that resistance and regulatory genes are not under CcaR control as opposed to pathway biosynthetic genes. In the intergenic cmcH-ccaR region, a small messenger RNA (mRNA) overlaps with the cmcH transcription terminator. Deletion of 688 bp of the intergenic region results in a strain, S. clavuligerus ΔRI, still able to produce cephamycin C and clavulanic acid but at levels 30-40% lower than the parental strain. Therefore, specific sequences in the intergenic region upstream of ccaR enhance the expression of ccaR but are not essential for its expression. Strains containing an additional ccaR gene integrated in the chromosome, S. clavuligerus pSET-PC, or multiple copies of ccaR expressed from the PglpF promoter, S. clavuligerus pAK23, were constructed. Fermentations of the pAK23 strain resulted in a 6.1-fold increase in specific cephamycin C production relative to the wild type. In the same experiments, qRT-PCR analysis of the cephamycin biosynthesis genes showed a 5.1-fold increase in ccaR expression and similar increases in expression of lat and cmcI, while expression of other cluster genes were increased in the order of 2- to 3-fold.


The first decade of antiretroviral therapy in Africa.
The past decade has seen remarkable progress in increasing access to antiretroviral therapy in resource-limited settings. Early concerns about the cost and complexity of treatment were overcome thanks to the efforts of a global coalition of health providers, activists, academics, and people living with HIV/AIDS, who argued that every effort must be made to ensure access to essential care when millions of lives depended on it. The high cost of treatment was reduced through advocacy to promote access to generic drugs; care provision was simplified through a public health approach to treatment provision; the lack of human resources was overcome through task-shifting to support the provision of care by non-physicians; and access was expanded through the development of models of care that could work at the primary care level. The challenge for the next decade is to further increase access to treatment and support sustained care for those on treatment, while at the same time ensuring that the package of care is continuously improved such that all patients can benefit from the latest improvements in drug development, clinical science, and public health.


Floating debris in the Mediterranean Sea.
Results from the first large-scale survey of floating natural (NMD) and anthropogenic (AMD) debris (>2 cm) in the central and western part of the Mediterranean Sea are reported. Floating debris was found throughout the entire study area with densities ranging from 0 to 194.6 items/km(2) and mean abundances of 24.9 AMD items/km(2) and 6.9 NMD items/km(2) across all surveyed locations. On the whole, 78% of all sighted objects were of anthropogenic origin, 95.6% of which were petrochemical derivatives (i.e. plastic and styrofoam). Maximum AMD densities (>52 items/km(2)) were found in the Adriatic Sea and in the Algerian basin, while the lowest densities (<6.3 items/km(2)) were observed in the Central Tyrrhenian and in the Sicilian Sea. All the other areas had mean densities ranging from 10.9 to 30.7 items/km(2). According to our calculations, more than 62 million macro-litter items are currently floating on the surface of the whole Mediterranean basin.


Alternative projections of the U.S. population.
The U.S. Bureau of the Census recently released a set of population projections that include middle and high projections that we argue are too conservative. The projections discount the possibility of future baby booms and assume slow rates of mortality decline and low levels of immigration. In this article we explore the impact on the size and age composition of the U.S. population of alternative scenarios of plausible fertility, mortality, and immigration assumptions. We conclude that (1) the Census Bureau's highest projection might be interpreted as a reasonable middle projection, (2) a reasonable high projection would yield a U.S. population in 2080 some 300 million persons larger than the Bureau's highest projection, with the population 85 and older more than twice the Bureau's greatest estimate, and (3) uncertainty about the pace of population growth is substantially greater than the Bureau's projections suggest.


Recruitment of the tinman homolog Nkx-2.5 by serum response factor activates cardiac alpha-actin gene transcription.
We recently showed that the cardiogenic homeodomain factor Nkx-2.5 served as a positive acting accessory factor for serum response factor (SRF) and that together they provided strong transcriptional activation of the cardiac alpha-actin promoter, depending upon intact serum response elements (SREs) (C. Y. Chen, J. Croissant, M. Majesky, S. Topouz, T. McQuinn, M. J. Frankovsky, and R. J. Schwartz, Dev. Genet. 19:119-130, 1996). As shown here, Nkx-2.5 and SRF collaborated to activate the endogenous murine cardiac alpha-actin gene in 10T1/2 fibroblasts by a mechanism in which SRF recruited Nkx-2.5 to the alpha-actin promoter. Activation of a truncated promoter consisting of the proximal alpha-actin SRE1 occurred even when Nkx-2.5 DNA-binding activity was blocked by a point mutation in the third helix of its homeodomain. Investigation of protein-protein interactions showed that Nkx-2.5 was bound to SRF in the absence of DNA in soluble protein complexes retrieved from cardiac myocyte nuclei but could also be detected in coassociated binding complexes on the proximal SRE1. Recruitment of Nkx-2.5 to an SRE depended upon SRF DNA-binding activity and was blocked by the dominant negative SRFpm1 mutant, which allowed for dimerization of SRF monomers but prevented DNA binding. Interactive regions shared by Nkx-2.5 and SRF were mapped to N-terminal/helix I and helix II/helix III regions of the Nkx-2.5 homeodomain and to the N-terminal extension of the MADS box. Our study suggests that physical association between Nkx-2.5 and SRF is one way that cardiac specified genes are activated in cardiac cell lineages.


[Clinical features, hormonal profile, and metabolic abnormalities of obese women with obese polycystic ovary syndrome].
To investigate and analyze the clinical presentation, hormonal profile, and metabolic abnormalities of obese women with polycystic ovary syndrome (PCOS). The data of the anthropometric measurements, clinical manifestations of hyperandrogenism, serum levels of luteinizing hormone (LH), follicle stimulating hormone (FSH), estradiol (E(2)), testosterone (T), prolactin (PRL), dehydro-epiandrosterone sulfate (DHEAS), sex-hormone-binding globulin (SGBG), and 17-oxyhydroprogesterone (17-OHP), fasting plasma glucose (FPG) and fasting insulin (FINS) detected after oral glucose tolerance test (OGTT), serum lipid levels, including total cholesterol (Chol), triglycerides (TG), high-density lipoprotein (HDL), and low-density lipoprotein (LDL), homeostasis model assessment (HOMA) and area under curve (AUC) so as to assess the insulin resistance (IR), free androgen index (FAI) to estimate the extent of hyperandrogenism, HOMA IS and DeltaI(30)/DeltaG(30) used to assess the function of islet beta cells, were collected from 192 women with PCOS, aged 24 +/- 6, that were divided into 2 groups according to the body mass index (BMI): Group A (n = 70) with the BMI > or = 25 kg.m(-2) and Group B (n = 122) with the BMI < 2 5 kg.m(-2), and 65 age-matched bilateral tubal block factor infertile women served as controls that were divided into 2 groups as well: Group C (n = 25) with the BMI > or = 25 kg.m(-2); and Group D (n = 79) with the BMI < 25 kg.m(-2), and underwent a cross-sectional study. (1) Clinical phenotype: The presence of obesity was 36.46% (70/192) of which 80.00% (56/70) was central obesity. The incidence of acanthosis nigricans was 17.18% (33/192), 35.71% in Group A and 6.56% in Group B. (P < 0.01). Groups A and C showed increased frequency of acanthosis nigricans compared with Group B. The value of FAI of Group A was 3.40 +/- 1.84, significantly higher than those of Group B (1.75 +/- 1.20) and Group C (1.65 +/- 0.90), (both P < 0.01). The LH/FSH ratio of Group B was 2.41 +/- 1.13, significantly higher than those of Groups A, C, and D (all P < 0.01). (2) Hormonal profile: The IR rate was 43.23% in the 192 patients, 82.86% in Group A and 20.49% in Group B. The LH and LH/FSH ratio were significantly higher in Group B than in Groups A, C, and D (all P < 0.01); T level was higher in Groups A and B than in Group C and D (all P < 0.05). SHBG was lower in Group A (108.70 +/- 81.35 nmol.L(-1)) and Group C (150.34 +/- 106.23 nmol.L(-1)) compared with Group B (192.49 +/- 98.30 nmol.L(-1)) and Group D (231.84 +/- 90.09 nmol.L(-1)) (P < 0.01 and P < 0.05). FAI level was 3.40 +/- 1.84 in Group A, significantly higher than those of Groups B (1.75 +/- 1.20), C (1.65 +/- 0.90), and D (0.84 +/- 0.45) (all P < 0.01). The FINS, TG, and HOMA IR of Groups A and C were all significantly higher than those of Groups B and D (all P < 0.01). The OGTT GAUC was significantly higher than those of Groups B, C, and D (P = 0.006, 0.028, and 0.031 respectively). (3) Metabolic profile: The prevalence of IR was 43.23% (83/192) with a higher prevalence rate in Group A (82.76%, 58/70) compared with Group B (20.49%, 25/122). The values of FINS, HOMA IR, GAUC, IAUC, and TG were all higher in Group A than in Group B (all P < 0.01). BMI and WHR were positively correlated with FAI and HOMA-IR (all P < 0.01), whereas negatively correlated with LH/FSH ratio (r = -0.345, -0.260, P < 0.01). There were no significant differences in HOMA-IS and DeltaI(30)/DeltaG(30) among these groups (all P > 0.05). Obese PCOS women have more severe hyperandrogenism, IR and hyperinsulinism than normal-weight PCOS women, which may have some health implications later in life.


Bevacizumab fails to treat temporal paraganglioma: discussion and case illustration.
Temporal paragangliomas are highly vascular tumors treated primarily by surgical resection. However, surgery to remove these tumors is associated with significant morbidity, including cranial nerve dysfunction. Interestingly, these tumors have been shown to express vascular endothelial growth factor (VEGF). A variety of tumors expressing VEGF and the VEGF receptor have been shown to reduce in size and vascularity when treated with the VEGF-specific antibody, bevacizumab (Avastin). We hypothesized that paragangliomas may be treated noninvasively with bevacizumab, either as a primary treatment or as a useful adjuvant to surgical resection or radiation. Thus, our aim was to evaluate the effects of bevacizumab on this patient's paraganglioma. A 36-year-old female presented to us with a 3 month history of positional dizziness, light-headedness, and left ear pulsatile tinnitus and hearing loss. She was found to have a temporal paraganglioma (glomus jugulare tumor) on imaging. Histopathology confirmed significant staining for VEGF. This patient was treated with bevacizumab prior to surgical treatment; radiographic imaging at 3 months, however, showed no significant response. We discuss possible reasons for treatment failure.


Nitrogen and sulphur relations in effecting yield and quality of cereals and oilseed crops.
Nitrogen and sulphur, both vital structural elements, are especially needed for the synthesis of proteins and oils. Investigations revealed the required application of sulphur is one half to one third the amount of nitrogen, and the ratio becomes narrower in mustard (Brassica juncea L.), followed by wheat and rice. The efficiency of an increased level of nitrogen required a proportionately higher amount of sulphur. A critical investigation on the effective utilization of applied vis-à-vis absorbed nitrogen in wheat and mustard envisaged accumulation of NO3-N in vegetative parts when sulphur remained proportionately low. Application of sulphur hastened the chemical reduction of absorbed NO3- for its effective utilization. The effect was more pronounced in mustard than in wheat. Easily available forms of sulphur, like ammonium sulphate and gypsum, as compared to pyrite or elemental sulphur, maintained adequate N to S ratio in rice, resulting in a reduction in the percent of unfilled grain, a major consideration in rice yield. A narrow N to S ratio, with both at higher levels, increased the oil content but raised the saponification value of the oil, a measure of free fatty acids. Whereas, a proportionately narrow N to S ratio at moderate dose resulted in adequately higher seed and oil yield with relatively low saponification value, associated with increased iodine value of the oil, indicating respectively low free fatty acids and higher proportion of unsaturated fatty acids, an index for better quality of the oil.


Purification and identification of G protein-coupled receptor protein complexes under native conditions.
G protein-coupled receptors (GPCRs) constitute the largest family of membrane receptors and are of major therapeutic importance. The identification of GPCR-associated proteins is an important step toward a better understanding of these receptors. However, current methods are not satisfying as only isolated receptor domains (intracellular loops or carboxyl-terminal tails) can be used as "bait." We report here a method based on tandem affinity purification coupled to mass spectrometry that overcomes these limitations as the entire receptor is used to identify protein complexes formed in living mammalian cells. The human MT(1) and MT(2) melatonin receptors were chosen as model GPCRs. Both receptors were tagged with the tandem affinity purification tag at their carboxyl-terminal tails and expressed in human embryonic kidney 293 cells. Receptor solubilization and purification conditions were optimized. The method was validated by the co-purification of G(i) proteins, which are well known GPCR interaction partners but which are difficult to identify with current protein-protein interaction assays. Several new and functionally relevant MT(1)- and MT(2)-associated proteins were identified; some of them were common to both receptors, and others were specific for each subtype. Taken together, our protocol allowed for the first time the purification of GPCR-associated proteins under native conditions in quantities suitable for mass spectrometry analysis.


A Study of Pap Smears in HIV-Positive and HIV-Negative Women from a Tertiary Care Center in South India.
The aim of this work was to study the spectrum of epithelial abnormalities on Pap smears of HIV-positive women categorized as per the Bethesda System of Reporting Cervical Cytology, to correlate them with CD4 lymphocyte counts, and to compare them with the spectrum of abnormalities seen in a HIV-negative control group. Study Design and Methodology: The present study was a 6-year retrospective study conducted in the Department of Pathology at Kasturba Medical College, Mangalore, which included 150 Pap smears from HIV-positive and HIV-negative women, respectively. The Pap-stained slides of the cases were retrieved and studied. The data collected were tabulated and analyzed. A statistical study was performed using SPSS software. The χ2 test was used to analyze the data and a p value < 0.05 was considered to be significant. Pap smear abnormalities were twice as high in HIV-infected women (12%) as compared with HIV-negative women (6%; p = 0.006, RR = 2). Negative for intraepithelial lesion/malignancy was the most common finding (88%), which was further subdivided into inflammatory, atrophic smear, non-specific, candidiasis, and bacterial vaginitis groups. The percentage of epithelial abnormalities was 12%, including: atypical squamous cells of undetermined significance, 5.55%; atypical squamous cells, cannot exclude HSIL, 16.66%; low-grade squamous intraepithelial lesion, 5.55%; high-grade squamous intraepithelial lesion (HSIL), 61.11%, and squamous cell carcinoma, 11.11%. The highest incidence of intraepithelial lesions in HIV-positive females was in the age group of 34-49 years. CD4 cell counts fell in the range of 200-500 cells/mm3 in most of the HIV-positive patients (68.75%), but was not found to be statistically significant. Routine Pap smear examination is advocated in women with HIV as the prevalence of epithelial cell abnormalities was found to be 12%, which was twice as high as compared to the HIV-negative control group. Although there was no correlation of epithelial cell abnormalities with CD4 counts, a higher rate of the cases with epithelial abnormalities were observed to have CD4 cell counts of 200-500 cells/mm3.


Acquisition of /s/ clusters in English-speaking children with phonological disorders.
Two member onset consonant clusters with /s/ as the first member (#sC onsets) behave differently from other double onset consonant clusters in English. Phonological explanations of children's consonant cluster production have been posited to predict children's speech acquisition. The aim of this study was to consider the role of the Sonority Sequencing Principle (SSP), homorganicity, headedness, and factorial typology in the explanation of productions of consonant clusters by children with phonological disorders. Thirty monolingual English-speaking children with phonological disorders produced words commencing with #sC: /s/+stop (/sT/), /s/+nasal (/sN/), /sl/, and /sw/. There was wide individual variation between participants. Overall, there was an increase in accuracy of consonant cluster production as the sonority indices of the second element of the cluster increased: there was significantly greater accuracy of /sl/ than /sN/ and significantly greater accuracy of /sN/ than /sT/. However, /sw/ targets only followed the SSP for phonologically accurate (but not phonetically accurate) productions. Correct renditions justified the binary grouping of the SSP-following vs SSP-violating separation. Homorganicity of the cluster was not a factor in determining relative accuracy of the targets. The percentage of correct renditions justified the /s/+[-continuant] vs /s/+[+continuant] binary separation. There were several counterexamples of predictions of the headedness approach. Thus, sonority and factorial typology were found to be useful in predicting /s/ consonant cluster acquisition of children with phonological disorders.


Evaluation of the effect of a chicken comb extract-containing supplement on cartilage and bone metabolism in athletes.
In a previous study, we revealed that a commercially available product of dietary supplement containing a chicken comb extract (CCE), which is rich in hyaluronan, not only relieves joint pain and other symptoms, but also potentially improves the balance of type II collagen degradation/synthesis in patients with knee osteoarthritis. Since soccer is one of the sports most likely to cause knee osteoarthritis (OA), we evaluated the effect of a CCE-containing supplement on cartilage and bone metabolism in athletes. Fourteen and 15 subjects (all midfielders) were randomly assigned to receive the test product (test group) and the dummy placebo containing only vehicle (placebo group), respectively, for 12 weeks. The daily oral intake of the CCE-containing test product clearly decreased the urinary levels of both C-terminal crosslinked telopeptides of cartilage-specific type II collagen (CTX-II) as a type II collagen degradation marker and the N-terminal telopeptides of bone-specific type I collagen (NTx) as a marker of bone resorption at 12 weeks after the initiation of the intervention. By contrast, no significant reduction was detected in the placebo group at any timepoint during the intervention. These observations indicate that the test product is effective in inhibiting, not only cartilage degradation, but also bone remodeling. Thus, the CCE-containing supplement may be useful for the management of joint health in athletes.