Role of extracellular proton-sensing OGR1 in regulation of insulin secretion and pancreatic β-cell functions.
Insulin secretion with respect to pH environments has been investigated for a long time but its mechanism remains largely unknown. Extracellular pH is usually maintained at around 7.4 and, its change has been thought to occur in non-physiological situations. Acidification takes place under ischemic and inflammatory microenvironments, where stimulation of anaerobic glycolysis results in the production of lactic acid. In addition to ionotropic ion channels, such as transient receptor potential V1 (TRPV1) and acid-sensing ion channels (ASICs), metabotropic proton-sensing G protein-coupled receptors (GPCRs) have also been identified recently as proton-sensing machineries. While ionotropic ion channels usually sense strong acidic pH, proton-sensing GPCRs sense pH of 7.6 to 6.0 and have been shown to mediate a variety of biological actions in neutral and mildly acidic pH environments. Studies with receptor knockout mice have revealed that proton-sensing receptors, including ovarian cancer G protein-coupled receptor 1 (OGR1), a proton-sensing GPCRs, play a role in the regulation of insulin secretion and glucose metabolism under physiological conditions. Small molecule 3,5-disubstituted isoxazoles have recently been identified as OGR1 agonists working at neutral pH and have been shown to stimulate pancreatic β-cell differentiation and insulin synthesis. Thus, proton-sensing OGR1 may be an important player for insulin secretion and a potential target for improving β-cell function.


IN VITRO ACTIVITY OF VACCINIUM MACROCARPON (CRANBERRY) ON URINARY TRACT PATHOGENS IN UNCOMPLICATED URINARY TRACT INFECTION.
Urinary tract infection is the most common bacterial infection in the community, mainly caused by Escherichia coli (E coli). Due to its high incidence and recurrence, problems are faced in the treatment with antibiotics. Cranberry being herbal remedy have long been the focus of interest for their beneficial effects in preventing urinary tract infections. This study was conducted to analyse in vitro activity of cranberry (Vaccinium macrocarpon) on uropathogenic E coli in uncomplicated urinary tract infections. In this laboratory based single group experimental study, anti-bacterial activity of Vaccinium macrocarpon concentrate on urinary tract E coli was investigated, in vitro. Ninety-six culture positive cases of different uropathogens were identified. Vaccinium macrocarpon concentrate at different concentrations was prepared in distilled water and put in wells punched in nutrient agar. E coli isolates were inoculated on the plates and incubated at 37 °C for 24 hours. A citric acid solution of the same pH as that of Vaccinium macrocarpon was used and put in a well on the same plate to exclude the effect of pH. A total of 35 isolates of E coli were identified out of 96 culture positive specimens of urine and found sensitive to Vaccinium macrocarpon (p<0.000). Results revealed that Vaccinium macrocarpon has antibacterial effect against E coli. Furthermore the antibacterial activity of Vaccinium macrocarpon has dose response relationship. Acidic nature of Vaccinium macrocarpon due to its pH is not contributory towards its antibacterial effect. Vaccinium macrocarpon concentrate may be used in urinary tract infection caused by E coli.


Synthesis, characterization, antibacterial activity and cytotoxicity of hollow TiO2-coated CeO2 nanocontainers encapsulating silver nanoparticles for controlled silver release.
Biomaterials as implants are being applied more extensively in medicine due to their on-going development and associated improvements, and the increase in human life expectancy. Nonetheless, biomaterial-related infections, as well as propagating bacterial resistance, remain significant issues. Therefore, there is a growing interest for silver-based drugs because of their efficient and broad-range antimicrobial activity and low toxicity to humans. Most newly-developed silver-based drugs have an extremely fast silver-ion release, increasing adverse biological impact to the surrounding tissue and achieving only short-term antimicrobial activity. Nanoencapsulation of these drugs is hypothesized as beneficial for controlling silver release, and thus is the aim of the present study. Initially, an amorphous or crystalline (anatase) titania (TiO2) coating was synthesized around silver nanoparticle-containing (AgNP) ceria (CeO2) nanocontainers using a sonication method forming AgNP/CeO2/TiO2 nanocontainers. These nanocontainers were characterized by high-resolution transmission electron microscopy, scanning electron microscopy, powder X-ray diffraction, gas sorption experiments and energy-dispersive X-ray spectroscopy. Silver release, monitored by using inductively coupled plasma optical emission spectroscopy, showed that these containers prevented silver release in water at neutral pH, and released the silver in concentrated nitric acid solution (pH = 1.1). The AgNP/CeO2/TiO2 nanocontainers showed an antibacterial activity against E. coli, however a concentration-dependent cytotoxicity towards a model epithelial barrier cell type (A549 cells) was observed. These nanocontainers offer the concept of potentially controlling silver delivery for the prevention of implant-associated infections.


A pragmatic randomised controlled trial of the effectiveness and cost-effectiveness of 'PhysioDirect' telephone assessment and advice services for physiotherapy.
As a result of long delays for physiotherapy for musculoskeletal problems, several areas in the UK have introduced PhysioDirect services in which patients telephone a physiotherapist for initial assessment and treatment advice. However, there is no robust evidence about the effectiveness, cost-effectiveness or acceptability to patients of PhysioDirect. To investigate whether or not PhysioDirect is equally as clinically effective as and more cost-effective than usual care for patients with musculoskeletal (MSK) problems in primary care. Pragmatic randomised controlled trial to assess equivalence, incorporating economic evaluation and nested qualitative research. Patients were randomised in 2 : 1 ratio to PhysioDirect or usual care using a remote automated allocation system at the level of the individual, stratifying by physiotherapy site and minimising by sex, age group and site of MSK problem. For the economic analysis, cost consequences included NHS and patient costs, and the cost of lost production. Cost-effectiveness analysis was carried out from the perspective of the NHS. Interviews were conducted with patients, physiotherapists and their managers. Four community physiotherapy services in England. Adults referred by general practitioners or self-referred for physiotherapy for a MSK problem. Patients allocated to PhysioDirect were invited to telephone a senior physiotherapist for initial assessment and advice using a computerised template, followed by face-to-face care when necessary. Patients allocated to usual care were put on to a waiting list for face-to-face care. Primary outcome was the Short Form questionnaire-36 items, version 2 (SF-36v2) Physical Component Score (PCS) at 6 months after randomisation. Secondary outcomes included other measures of health outcome [Measure Yourself Medical Outcomes Profile, European Quality of Life-5 Dimensions (EuroQol health utility measure, EQ-5D), global improvement, response to treatment], wait for treatment, time lost from work and usual activities, patient satisfaction. Data were collected by postal questionnaires at baseline, 6 weeks and 6 months, and from routine records by researchers blind to allocation. A total of 1506 patients were allocated to PhysioDirect and 743 to usual care. Patients allocated to PhysioDirect had a shorter wait for treatment than those allocated to usual care [median 7 days vs 34 days; arm-time ratio 0.32, 95% confidence interval (CI) 0.29 to 0.35] and had fewer non-attended face-to-face appointments [incidence rate ratio 0.55 (95% CI 0.41 to 0.73)]. The primary outcome at 6 months' follow-up was equivalent between PhysioDirect and usual care [mean PCS 43.50 vs 44.18, adjusted difference in means -0.01 (95% CI -0.80 to 0.79)]. The secondary measures of health outcome all demonstrated equivalence at 6 months, with slightly greater improvement in the PhysioDirect arm at 6 weeks' follow-up. Patients were equally satisfied with access to care but slightly less satisfied overall with PhysioDirect compared with usual care. NHS costs (physiotherapy plus other relevant NHS costs) per patient were similar in the two arms [PhysioDirect £ 198.98 vs usual care £ 179.68, difference in means £ 19.30 (95% CI -£ 37.60 to £ 76.19)], while QALYs gained were also similar [difference in means 0.007 (95% CI -0.003 to 0.016)]. Incremental cost per QALY gained was £ 2889. The probability that PhysioDirect was cost-effective at a £ 20,000 willingness-to-pay threshold was 88%. These conclusions about cost-effectiveness were robust to sensitivity analyses. There was no evidence of difference between trial arms in cost to patients or value of lost production. No adverse events were detected. Providing physiotherapy via PhysioDirect is equally clinically effective compared with usual waiting list-based care, provides faster access to treatment, appears to be safe, and is broadly acceptable to patients. PhysioDirect is probably cost-effective compared with usual care.


The sequence of the M RNA of an isolate of La Crosse virus.
The middle-size (M) genomic RNA of a New York State, U.S.A. isolate of La Crosse (LAC) virus has been cloned by a random priming procedure and its nucleotide sequence determined by the dideoxy method. The RNA was found to be 4526 nucleotides in length and to have a base composition of 34.2% U, 27.8% A, 20.6% C and 17.4% G. There is a single, long open reading frame in the viral complementary RNA that contains sufficient information to code for a protein of 1441 amino acids. In these respects, as in many others, the LAC virus M RNA and its encoded protein were very similar, if not identical, to those previously reported by other investigators for the closely related snowshoe hare virus. The M RNAs of the two viruses show 79% nucleotide sequence homology and 89% homology in the amino acid sequence of their encoded proteins. Several algorithms for predicting surface residues, as well as the Chou-Fasman rules for predicting secondary structure, were used to compare the LAC virus and snowshoe hare virus M gene proteins. These analyses identified 39 sites on the proteins as those most likely to be linear antigenic determinants that might contribute to the differences between the two viruses.


Impacts of entropy generation in second-grade fuzzy hybrid nanofluids on exponentially permeable stretching/shrinking surface.
The present investigation aims to use entropy analysis to analyze the unsteady Magnetohydrodynamic (MHD) flow in a second-grade fuzzy hybrid [Formula: see text] nanofluid over an exponentially shrinking/stretching surface. The model for hybridization of the mixture of alumina [Formula: see text] and copper (Cu) nanoparticles in the sodium alginate (SA) base fluid under heat source/sink, nonlinear thermal radiation, and viscous dissipation. The fundamental partial differential equations (PDEs) are simplified using an appropriate similarity conversion to generate the ordinary differential equations (ODEs). The analytical computation occurs in the MATHEMATICA program implementing the homotopy analysis method (HAM). In terms of code validity, our results are preferable to previous findings. The features of several parameters against the velocity, surface friction coefficient, entropy, temperature, and Nusselt number are described through graphs. According to our findings, the rise in the Brinkman and Reynolds numbers enhanced the total entropy of the system. Furthermore, the nanoparticle volume fraction and viscus dissipation magnifies the fluid temperature while retards the flow profile throughout the domain. Fluid velocity declined due to the Lorentz force using magnetic impact applications. The imprecision of nanofluid and hybrid nanofluid volume fractions was modelled as a triangular fuzzy number (TFN) [0%, 1%, 2%] for comparison. The double parametric approach was applied to deal with the fuzziness of the associated fuzzy parameters. The nonlinear ODEs convert into fuzzy differential equations (FDEs) and use HAM for the fuzzy solution. From our observation, the hybrid nanofluid displays the maximum heat transfer compared to nanofluids. This important contribution will support industrial growth, particularly in the processing and manufacturing sectors. The percentage increase in skin friction factor is 18.3 and 15.0 when [Formula: see text] and [Formula: see text] take input in the ranges of 0 ≤ [Formula: see text] ≤ 0.8 and 0 ≤ [Formula: see text] ≤ 1, respectively.


A review on locomotion robophysics: the study of movement at the intersection of robotics, soft matter and dynamical systems.
Discovery of fundamental principles which govern and limit effective locomotion (self-propulsion) is of intellectual interest and practical importance. Human technology has created robotic moving systems that excel in movement on and within environments of societal interest: paved roads, open air and water. However, such devices cannot yet robustly and efficiently navigate (as animals do) the enormous diversity of natural environments which might be of future interest for autonomous robots; examples include vertical surfaces like trees and cliffs, heterogeneous ground like desert rubble and brush, turbulent flows found near seashores, and deformable/flowable substrates like sand, mud and soil. In this review we argue for the creation of a physics of moving systems-a 'locomotion robophysics'-which we define as the pursuit of principles of self-generated motion. Robophysics can provide an important intellectual complement to the discipline of robotics, largely the domain of researchers from engineering and computer science. The essential idea is that we must complement the study of complex robots in complex situations with systematic study of simplified robotic devices in controlled laboratory settings and in simplified theoretical models. We must thus use the methods of physics to examine both locomotor successes and failures using parameter space exploration, systematic control, and techniques from dynamical systems. Using examples from our and others' research, we will discuss how such robophysical studies have begun to aid engineers in the creation of devices that have begun to achieve life-like locomotor abilities on and within complex environments, have inspired interesting physics questions in low dimensional dynamical systems, geometric mechanics and soft matter physics, and have been useful to develop models for biological locomotion in complex terrain. The rapidly decreasing cost of constructing robot models with easy access to significant computational power bodes well for scientists and engineers to engage in a discipline which can readily integrate experiment, theory and computation.


The Brf1 and Bdp1 subunits of transcription factor TFIIIB bind to overlapping sites in the tetratricopeptide repeats of Tfc4.
The RNA polymerase III initiation factor TFIIIB is assembled onto DNA through interactions involving the Tfc4 subunit of the assembly factor TFIIIC and two subunits of TFIIIB, Brf1 and Bdp1. Tfc4 contains two arrays of tetratricopeptide repeats (TPRs), each of which provides a binding site for Brf1. Dominant mutations in the ligand binding channel of the first TPR array, TPRs1-5, and on the back side of this array, increase Brf1 binding by Tfc4. Here we examine the biological importance of the second TPR array, TPRs6 -9. Radical mutations at phylogenetically conserved residues in the ligand binding channel of TPRs6 -9 impair pol III reporter gene transcription. Biochemical studies on one such mutation, L469K in TPR7, revealed a defect in the recruitment of Brf1 into TFIIIB-TFIIIC-DNA complexes and diminished the direct interaction between Tfc4 and Brf1. Multicopy suppression analysis implicates TPR9 in Brf1 binding and TPRs7 and 8 in binding to more than one ligand. Indeed, the L469K mutation also decreased the binding affinity for Bdp1 incorporation into TFIIIB-TFIIIC-DNA complexes and inhibited binary interactions between Bdp1 and Tfc4. The Bdp1 binding domain in Tfc4 was mapped to TPRs1-9, a domain that contains both TPR arrays and thus overlaps two of the known binding sites for Brf1. The properties of the L469K mutation identify both Brf1 and Bdp1 as ligands for the second TPR array.


MDMA-induced indifference to negative sounds is mediated by the 5-HT2A receptor.
MDMA has been shown to induce feelings of sociability, a positive emotional bias and enhanced empathy. While previous research has used only visual emotional stimuli, communication entails more than that single dimension and it is known that auditory information is also crucial in this process. In addition, it is, however, unclear what the neurobiological mechanism underlying these MDMA effects on social behaviour is. Previously, studies have shown that MDMA-induced emotional excitability and positive mood are linked to the action on the serotonin (5-HT) 2A receptor. The present study aimed at investigating the effect of MDMA on processing of sounds (Processing of Affective Sounds Task (PAST)) and cognitive biases (Approach-Avoidance Task (AAT)) towards emotional and social stimuli and the role of 5-HT2A receptor in these effects. Twenty healthy recreational users entered a 2 × 2, placebo-controlled, within-subject study with ketanserin (40 mg) as pre-treatment and MDMA (75 mg) as treatment. Behavioural (PAST, AAT) measures were conducted 90 min after treatment with MDMA, respectively, 120 min after ketanserin. Self-report mood measures and oxytocin concentrations were taken at baseline and before and after behavioural tests. Findings showed that MDMA reduced arousal elicited by negative sounds. This effect was counteracted by ketanserin pre-treatment, indicating involvement of the 5-HT2 receptor in this process. MDMA did not seem to induce a bias towards emotional and social stimuli. It increased positive and negative mood ratings and elevated oxytocin plasma concentrations. The reduction in arousal levels when listening to negative sounds was not related to the elevated subjective arousal. It is suggested that this decrease in arousal to negative stimuli reflects potentially a lowering of defences, a process that might play a role in the therapeutic process.


Perceptual cues used by listeners to discriminate fluent from nonfluent narrative discourse.
BACKGROUND: Language fluency is a common diagnostic marker for discriminating among aphasia subtypes and improving clinical inference about site of lesion. Nevertheless, fluency remains a subjective construct that is vulnerable to a number of potential sources of variability, both between and within raters. Moreover, this variability is compounded by distinct neurological aetiologies that shape the characteristics of a narrative speech sample. Previous research on fluency has focused on characteristics of a particular patient population. Less is known about the ways that raters spontaneously weigh different perceptual cues when listening to narrative speech samples derived from a heterogeneous sample of brain-damaged adults. AIM: We examined the weighted contribution of a series of perceptual predictors that influence listeners' judgements of language fluency among a diverse sample of speakers. Our goal was to sample a range of narrative speech representing most fluent (i.e., healthy controls) to potentially least nonfluent (i.e., left inferior frontal lobe stroke). METHODS #ENTITYSTARTX00026; PROCEDURES: Three raters blind to patient diagnosis made forced choice judgements of fluency (i.e., fluent or nonfluent) for 61 pseudorandomly presented narrative speech samples elicited by the BDAE Cookie Theft picture. Samples were collected from a range of clinical populations, including patients with frontal and temporal lobe pathologies and non-brain-damaged speakers. We conducted a logistic regression analysis in which the dependent measure was the majority judgement of fluency for each speech sample (i.e., fluent or non-fluent). The statistical model contained five predictors: speech rate, syllable type token ratio, speech productivity, audible struggle, and filler ratio. OUTCOMES #ENTITYSTARTX00026; RESULTS: This statistical model fit the data well, discriminating group membership (i.e., fluent or nonfluent) with 95.1% accuracy. The best step of the regression model included the following predictors: speech rate, speech productivity, and audible struggle. Listeners were sensitive to different weightings of these predictors. CONCLUSIONS: A small combination of perceptual variables can strongly discriminate whether a listener will assign a judgement of fluent versus nonfluent. We discuss implications for these findings and identify areas of potential future research towards further specifying the construct of fluency among adults with acquired speech and language disorders.


A retrospective study of human African Trypanosomiasis in three Malawian Districts.
Human African Trypanosomiasis (HAT) is a neglected tropical disease (NTD). Surveillance in many of the endemic areas is often inadequate. Up to date information on the HAT situation in Malawi, where the disease is endemic in some districts, provides opportunity to raise the profile of the disease and interest in prevention and control. A retrospective study was conducted in three Malawian districts: Nkhotakota, Rumphi and Kasungu to describe the prevalence of HAT. Hospital laboratory registers from January 2000 to December 2006 were used. The calculated annual district prevalence of Trypanosomiasis ranged from 0.29 cases per 100,000 population in 2000, to 0.58 cases per 100,000 population in 2003. Nkhotakota District had the highest case detection rate of trypanosomiasis of 16.56 cases per 100,000 in 2002 and the lowest rate in 2004 of 5.23 cases per 100,000. From 2004 onwards a decline in cases detected was observed. In Rumphi district the highest number of cases (17.67 cases per 100,000 population) was identified in 2003 and the lowest rate of 1.29 cases per 100,000 in 2001. The rate (17.67 cases per 100,000) found in 2003 represented a 5-fold increase of 2002 (3.02 cases per 100,000). In Kasungu the detection rate ranged from 0 per 100,000 in 2001, 2003 and 2004 to 0.99 cases per 100,000 in 2005. The number of cases in this district has remained low including in 2006, when a detection rate of 0.16 cases per 100,000 was observed. HAT is endemic in selected districts of Malawi. There is need to explore the feasibility of active disease surveillance and the establishment of permanent preventive and control measures.


HLA-A SNPs and amino acid variants are associated with nasopharyngeal carcinoma in Malaysian Chinese.
Nasopharyngeal carcinoma (NPC) arises from the mucosal epithelium of the nasopharynx and is constantly associated with Epstein-Barr virus type 1 (EBV-1) infection. We carried out a genome-wide association study (GWAS) of 575,247 autosomal SNPs in 184 NPC patients and 236 healthy controls of Malaysian Chinese ethnicity. Potential association signals were replicated in a separate cohort of 260 NPC patients and 245 healthy controls. We confirmed the association of HLA-A to NPC with the strongest signal detected in rs3869062 (p = 1.73 × 10(-9)). HLA-A fine mapping revealed associations in the amino acid variants as well as its corresponding SNPs in the antigen peptide binding groove (p(HLA-A-aa-site-99) = 3.79 × 10(-8), p(rs1136697) = 3.79 × 10(-8)) and T-cell receptor binding site (p(HLA-A-aa-site-145) = 1.41 × 10(-4), p(rs1059520) = 1.41 × 10(-4)) of the HLA-A. We also detected strong association signals in the 5'-UTR region with predicted active promoter states (p(rs41545520) = 7.91 × 10(-8)). SNP rs41545520 is a potential binding site for repressor ATF3, with increased binding affinity for rs41545520-G correlated with reduced HLA-A expression. Multivariate logistic regression diminished the effects of HLA-A amino acid variants and SNPs, indicating a correlation with the effects of HLA-A*11:01, and to a lesser extent HLA-A*02:07. We report the strong genetic influence of HLA-A on NPC susceptibility in the Malaysian Chinese.


Hyperhomocysteinaemia as a risk factor for venous thrombosis: an update of the current evidence.
Classical homocystinuria is associated with arterial vascular diseases and venous thrombosis. In the last decade, many studies, including some prospective studies, have been published indicating that moderate hyperhomocysteinaemia is also a risk factor for venous thrombosis. The 677C>T mutation in the methylenetetrahydrofolate reductase (MTHFR) gene is an important cause of mild hyperhomocysteinaemia. Recent metaanalyses show an elevated risk of venous thrombosis for subjects with the TT-genotype. Based on the concept of 'Mendelian randomisation', this observation supports the hypothesis that hyperhomocysteinaemia is a causal risk factor for venous thrombosis. The results of one homocysteine-lowering trial regarding venous thrombosis are awaited at the end of 2003. In this paper the current evidence for hyperhomocysteinaemia as a risk factor for venous thrombosis is being discussed.


Simple fabrication of one-dimensional metal nanostructures and their application for SERS analysis.
This review highlights work using the author's method for the preparation of highly aligned metallic nanofibers with one-dimensional aggregates of metal nanoparticles (MNPs) and their utilization in surface enhanced Raman scattering (SERS) analysis. The preparation method, which is based on the process of evaporation-induced self-assembly with DNA and a drying front movement, eliminates the need for lithography and an external field; it is also fast, cheap and easy. Dark-field scattering spectroscopy was used to study the strong plasmon coupling of MNPs in metallic nanofibers. Furthermore, Raman spectral imaging of the metallic nanofibers revealed the existence of intense hot spots localized along their axes, which played a significant role in the intensity of SERS signals from DNA bases and rhodamine B in the metallic nanofibers. Our results demonstrate the use of evaporation-induced self-assembly with DNA as a straightforward method to produce the one-dimensional coupling of localized plasmons with a longer scale, and to facilitate the fabrication of optical sensor chips for single-molecule detection via SERS.


Efficient metal-ion catalyzed template-directed oligonucleotide synthesis.
The Pb2+ and Zn2+ ions are efficient catalysts for the polycytidylic acid-directed polymerization of an activated guanylic acid derivative, guanosine 5'-phosphorimidazolide. The products include oligomers of 30 to 40 units in length. The nucleotide residues are predominantly 2'-5' linked when Pb2+ is the catalyst, and predominantly 3'-5' linked in the presence of Zn2+. The significance of these results in the context of the prebiotic evolution of RNA polymerase is discussed.


Comparison of treatment and outcomes for patients with acute myocardial infarction in Minneapolis/St. Paul, Minnesota, and Göteborg, Sweden.
Treatment of acute myocardial infarction (AMI) is changing, and differences in medical practice are observed within and between countries on the basis of local practice patterns and available technology. These differing approaches provide an opportunity to evaluate medical practice and outcomes at the population level. The primary aim of this study was to compare medical care in patients hospitalized with AMI in 2 large cities in Sweden and the United States. A secondary aim was to compare medical outcomes. All resident patients (age range, 30-74 years) hospitalized with AMI in Göteborg, Sweden (1995-1996), and a representative population-based sample of all patients with AMI in Minneapolis/St. Paul, Minn (1995). Patients with AMI in Göteborg (GB) were older than patients in Minneapolis/St. Paul (MSP), but fewer patients in GB had a prior history of cardiovascular disease. During the AMI admission, coronary angiography, percutaneous coronary angioplasty (PTCA), and coronary artery bypass grafting (CABG) were performed twice as frequently in MSP than in GB. Echocardiogram and exercise testing were more frequently performed in GB. During hospitalization, beta-blockers were more frequently prescribed in GB, whereas calcium channel blockers, long- and short-acting nitrates, intravenous nitroglycerine, digitalis, aspirin, oral anticoagulants, heparin, and lidocaine were significantly more common in MSP. Thrombolysis, acute PTCA, ACE inhibitors, and diuretics were similar. Reinfarction was higher in men in GB (4% vs 1%, P <.009) and women in GB (3% vs 1%, P = not significant). On discharge, beta-blockers and diuretics were prescribed significantly more often in GB, whereas calcium channel blockers, nitrates, and digitalis were prescribed more often in MSP. Aspirin and ACE inhibitors had similar usage rates. Despite these diagnostic and treatment contrasts, there were no differences in mortality rate at 30 days or after 3 years of follow-up after risk-adjusting for patient baseline differences. Comparing patients hospitalized with AMI in MSP and GB, we found marked differences in medical care, with invasive strategies more likely to be used in MSP. This may be the result of historical practice patterns, the healthcare system, and healthcare financing differences. Despite these differences, short- or long-term mortality rates were identical.


Phosphorylation-induced structural changes in smooth muscle myosin regulatory light chain.
We have performed complementary time-resolved fluorescence resonance energy transfer (TR-FRET) experiments and molecular dynamics (MD) simulations to elucidate structural changes in the phosphorylation domain (PD) of smooth muscle regulatory light chain (RLC) bound to myosin. PD is absent in crystal structures, leaving uncertainty about the mechanism of regulation. Donor-acceptor pairs of probes were attached to three site-directed di-Cys mutants of RLC, each having one Cys at position 129 in the C-terminal lobe and the other at position 2, 3, or 7 in the N-terminal PD. Labeled RLC was reconstituted onto myosin subfragment 1 (S1). TR-FRET resolved two simultaneously populated structural states of RLC, closed and open, in both unphosphorylated and phosphorylated biochemical states. All three FRET pairs show that phosphorylation shifts the equilibrium toward the open state, increasing its mol fraction by approximately 20%. MD simulations agree with experiments in remarkable detail, confirming the coexistence of two structural states, with phosphorylation shifting the system toward the more dynamic open structural state. This agreement between experiment and simulation validates the additional structural details provided by MD simulations: In the closed state, PD is bent onto the surface of the C-terminal lobe, stabilized by interdomain salt bridges. In the open state, PD is more helical and straight, resides farther from the C-terminal lobe, and is stabilized by an intradomain salt bridge. The result is a vivid atomic-resolution visualization of the first step in the molecular mechanism by which phosphorylation activates smooth muscle.


GPS signal reception under snow cover: a pilot study establishing the potential usefulness of GPS in avalanche search and rescue operations.
Avalanches are one of the major threats to life in high-mountain terrain and account every year for approximately 150 accidents causing injury or death in the United States alone. Every year avalanches cause significant property damages and a death toll of approximately 15 people in the United States. The specific characteristic of the avalanche accident is the extreme importance of getting to the buried victim as soon as, possible to improve survival. Approximately 40% of all buried victims survive 1 hour, and only about 20% survive 2 hours. Newer studies from Europe indicate that the initial survival probability is 92% at 15 minutes, 30% at 35 minutes, 27% at 90 minutes, and finally drops to 3% at 130 minutes. Unless prompt and efficient search and rescue are ensured, the prospect of buried victims is rather grim. Many tools have been used in the past to aid in retrieving buried victims including the avalanche cord, probing techniques, and in more recent time, the use of electronic beacon devices that allow search teams to locate the buried victim. The advent of satellite navigational aids (GPS, GLONASS) makes it possible to determine one's position with remarkable accuracy. We studied the degree to which the GPS satellite signal could penetrate through snow and be received by a commercially available GPS receiver. This information may lead to the development of an additional tool for precise and quick localization of buried victims in avalanche accidents and thus may substantially improve their survival by shortening the search time at the accident site. In this study we used a Motorola Traxar six-channel GPS receiver with amplifier unit connected to an antenna by means of a shielded coaxial cable. The antenna was buried under incremental covers of compact snow, and the reception of the GPS signal was measured at each burial depth: 5 cm, 15 cm, 25 cm, 35 cm, 45 cm, 55 cm, 1 m, and 1.5 m. The variables that were measured included signal quality, number of satellites received by the receiver, and their respective signal strength. A reference reading was taken from the GPS receiver above the testing site before measurements under snow cover were started. The satellite signals were received with good quality and precise readings up to a burial depth of 1 m. Under 1.5 m of snow the receiver was able to lock on only one satellite, making a positional reading impossible. The reception of the GPS signals under snow cover is possible and warrants further study directed toward the development of a search and rescue device using this technology.


Predicting small molecule fluorescent probe localization in living cells using QSAR modeling. 1. Overview and models for probes of structure, properties and function in single cells.
Small molecule fluorochromes (synonyms: biosensors, chemosensors, fluorescent probes, vital stains) are widely used to investigate the structure, composition, physicochemical properties and biological functions of living cells, tissues and organisms. Selective entry and accumulation within particular cells and cellular structures are key processes for achieving these diverse objectives. Despite the complexities, probes routinely are applied using standard protocols, often without experimenter awareness of what factors that control accumulation and localization. The mechanisms of many such selective accumulations, however, now are known. Moreover, the influence of physicochemical properties of probes on their uptake and localization often can be defined numerically, hence predicted, using quantitative structure activity relations (QSAR) models with its required numerical structure parameters (or "descriptors"). The state of the art of this approach is described. Available QSAR models are summarized for uptake into cells and localization in the cytosol, endoplasmic reticulum, generic biomembranes, Golgi apparatus, lipid droplets, lysosomes/endosomes, mitochondria, eukaryotic nuclei (histones and DNA), plasma membrane, and ribosomal RNA (cytoplasmic and nucleolar). Integration of such core models to both aid understanding and troubleshooting of current fluorescent probes and to assist the design of novel probes is outlined and illustrated using case examples. Limitations and generic problems arising with this approach and comments on application of such approaches to xenobiotics other than probes, e.g., drugs and herbicides, together with a brief note about an alternative approach to prediction, are given.


Bactericidal efficacy of mobile ultraviolet-C disinfection devices in reducing contamination in biosafety laboratories.
Biosafety research requires a wide range of microorganisms and thorough disinfection to prevent laboratory infection is often required. Ultraviolet-C (UV-C) exposure reduces bacterial and viral concentrations. Therefore, in this study, we aimed to evaluate the efficacy of a mobile UV-C device as a non-contact disinfection strategy. The bactericidal efficacy of the UV-C device was determined based on log10 decreases in the relative abundances of bacterial indicators, including Escherichia coli, Staphylococcus aureus, Staphylococcus albus, and Pseudomonas aeruginosa at 0.5 and 1.0 m after irradiation for 30, 60, and 90 min. Next, the reduction of natural bacteria in air and on surface as a result of the UV-C device exposure in the laboratory were determined. Exposure to the UV-C disinfection device resulted in mean log10 decreases in microbial contamination of 3.55 and 5.85 following irradiation for 30 and 90 min, respectively, at a distance of 0.5 m. Further, P. aeruginosa and E. coli were the most and least sensitive to UV-C exposure, respectively. The bacterial load in air decreased by 65.53% after 60 min of irradiation, while those on surfaces decreased by 44.19% and 78.23% after 30 and 60 min of irradiation, respectively. The UV-C device effectively reduced bacterial load after irradiation for over 60 min. Further studies are encouraged to determine the effectiveness of the UV-C disinfection device in frequently occupied institutions, such as primary medical, health, and nursery, and its efficiency in infection control.


[Various forms of the celiac trunk and its anastomoses with the superior mesenteric artery].
Three rare varieties of upper abdominal arteries were compared with similar cases in the anatomical literature. An attempt was made to obtain a classification of the supernumerary branches of the celiac trunk and of the anastomoses between the celiac trunk and the superior mesenteric artery. One or more supernumerary branches of the celiac artery can be observed: (1) the superior mesenteric artery; (2) an accessory hepatic artery; (3) a posterior pancreatic artery; (4) a colic artery; (5) an accessory splenic artery; (6) a connecting branch to the superior mesenteric artery, and (7) an inferior phrenic artery. The following types of anastomoses between the celiac artery and the superior mesenteric artery can be distinguished: (1) direct connection; (2) anastomoses within the hepatic artery; (3) anatomoses following pre- or postnatal stenosis, and (4) the pancreatic arcades. For the first type the theory of TANDLER (longitudinal anastomosis) is abandoned. The development of the second type is as yet unresolved. In the case of the last two types a postembryonal formation is possible.


Zika virus restriction of host antioxidant response is mediated by intracellular NS1 and reveals its ability to upregulate Bach1 expression.
Zika virus (ZIKV), has gained global attention due to its association with severe disorders, including microcephaly and congenital Zika syndrome. We investigated the role of ZIKV nonstructural protein 1 (NS1) in altering the host's antioxidant response. Using a stable cell line expressing NS1, we found that NS1 significantly reduced the expression of antioxidant-related genes, including heme oxygenase 1 (HO-1), NAD(P)H quinone dehydrogenase 1 (NQO1), and sequestosome-1 (SQSTM1), which are regulated NRF2. Interestingly, this effect was attributed to increased expression of BACH1, a factor that competes with NRF2 for binding to certain antioxidant responsive elements (ARE). Thus, ZIKV NS1-mediated disruption of the antioxidant system is linked to BACH1 overexpression. These findings offer insights into ZIKV pathogenesis and suggest potential therapeutic strategies targeting the NRF2-BACH1 axis.


Family factors in adolescent problematic Internet gaming: A systematic review.
Background and aims Familial influences are known to affect the likelihood of an adolescent becoming a problem gamer. This systematic review examined some of the key findings in empirical research on family factors related to adolescent problem gaming. Methods A total of 14 studies in the past decade were evaluated. Family-related variables included: (a) parent status (e.g., socioeconomic status and mental health), (b) parent-child relationship (e.g., warmth, conflict, and abuse), (c) parental influence on gaming (e.g., supervision of gaming, modeling, and attitudes toward gaming), and (d) family environment (e.g., household composition). Results The majority of studies have focused on parent-child relationships, reporting that poorer quality relationships are associated with increased severity of problem gaming. The paternal relationship may be protective against problem gaming; therefore, prevention programs should leverage the support of cooperative fathers. Discussion The intergenerational effects of problem gaming require further attention, in light of adult gamers raising their children in a gaming-centric environment. Research has been limited by a reliance on adolescent self-report to understand family dynamics, without gathering corroborating information from parents and other family members. The very high rates of problem gaming (>10%) reported in general population samples raise concerns about the validity of current screening tools. Conclusions Interventions for adolescents may be more effective in some cases if they can address familial influences on problem gaming with the active co-participation of parents, rather than enrolling vulnerable adolescents in individual-based training or temporarily isolating adolescents from the family system.


Transcriptomic Analysis of Tobacco Plants in Response to Whitefly Infection.
The whitefly Bemisia tabaci is one of the most destructive pests worldwide, and causes tremendous economic losses. Tobacco Nicotiana tabacum serves as a model organism for studying fundamental biological processes and is severely damaged by whiteflies. Hitherto, our knowledge of how tobacco perceives and defends itself against whiteflies has been scare. In this study, we analyze the gene expression patterns of tobacco in response to whitefly infestation. A total of 244 and 2417 differentially expressed genes (DEGs) were identified at 12 h and 24 h post whitefly infestation, respectively. Enrichment analysis demonstrates that whitefly infestation activates plant defense at both time points, with genes involved in plant pattern recognition, transcription factors, and hormonal regulation significantly upregulated. Notably, defense genes are more intensely upregulated at 24 h post infestation than at 12 h, indicating an increased immunity induced by whitefly infestation. In contrast, genes associated with energy metabolism, carbohydrate metabolism, ribosomes, and photosynthesis are suppressed, suggesting impaired plant development. Taken together, our study provides comprehensive insights into how plants respond to phloem-feeding insects, and offers a theoretical basis for better research on plant-insect interactions.


Physical activity after diagnosis and risk of prostate cancer progression: data from the cancer of the prostate strategic urologic research endeavor.
Vigorous activity after diagnosis was recently reported to be inversely associated with prostate cancer-specific mortality. However, men with metastatic disease may decrease their activity due to their disease; thus, a causal interpretation is uncertain. We therefore prospectively examined vigorous activity and brisk walking after diagnosis in relation to risk of prostate cancer progression, an outcome less susceptible to reverse causation, among 1,455 men diagnosed with clinically localized prostate cancer. Cox proportional hazards regression was used to examine vigorous activity, nonvigorous activity, walking duration, and walking pace after diagnosis and risk of prostate cancer progression. We observed 117 events (45 biochemical recurrences, 66 secondary treatments, 3 bone metastases, 3 prostate cancer deaths) during 2,750 person-years. Walking accounted for nearly half of all activity. Men who walked briskly for 3 h/wk or more had a 57% lower rate of progression than men who walked at an easy pace for less than 3 h/wk (HR = 0.43; 95% CI: 0.21-0.91; P = 0.03). Walking pace was associated with decreased risk of progression independent of duration (HR brisk vs. easy pace = 0.52; 95% CI: 0.29-0.91; P(trend) = 0.01). Few men engaged in vigorous activity, but there was a suggestive inverse association (HR ≥3 h/wk vs. none = 0.63; 95% CI: 0.32-1.23; P(trend) = 0.17). Walking duration and total nonvigorous activity were not associated with risk of progression independent of pace or vigorous activity, respectively. Brisk walking after diagnosis may inhibit or delay prostate cancer progression among men diagnosed with clinically localized prostate cancer.


Proliferative signaling by store-operated calcium channels opposes colon cancer cell cytostasis induced by bacterial enterotoxins.
Guanylyl cyclase C and accumulation of cGMP induced by bacterial heat-stable enterotoxins (STs) promote colon cancer cell cytostasis, serving as a tumor suppressor in intestine. Conversely, capacitative calcium entry through store-operated calcium channels (SOCs) is a key signaling mechanism that promotes colon cancer cell proliferation. The present study revealed that proliferative signaling by capacitative calcium entry through SOCs opposes and is reciprocally coupled to cytostasis mediated by guanylyl cyclase C in T84 human colon carcinoma cells. Elimination of capacitative calcium entry employing 2-aminoethoxydiphenylborate (2-APB), a selective inhibitor of SOCs, potentiated cytostasis induced by ST. Opposition of ST-induced cytostasis by capacitative calcium entry reflects reciprocal inhibition of guanylyl cyclase C signaling. Calcium entry through SOCs induced by the calcium-ATPase inhibitor thapsigargin or the receptor agonists UTP or carbachol inhibited guanylyl cyclase C-dependent cGMP accumulation. This effect was mimicked by the calcium ionophore ionomycin and blocked by 2-APB and intracellular 1,2-bis(o-amino-5,5'-dibromophenoxy)ethane-N,N,N',N'-tetraacetic acid-acetoxymethyl ester (BAPTA-AM), a chelator of calcium. Moreover, regulation by capacitative calcium entry reflected ligand-dependent sensitization of guanylyl cyclase C to inhibition by that cation. Although basal catalytic activity was refractory, ST-stimulated guanylyl cyclase C was inhibited by calcium, which antagonized binding of magnesium to allosteric sites required for receptor-effector coupling. These observations demonstrate that reciprocal regulation of guanylyl cyclase C signaling by capacitative calcium entry through SOCs represents one limb of a coordinated mechanism balancing colon cancer cell proliferation and cytostasis. They suggest that combining guanylyl cyclase C agonists and SOC inhibitors offers a novel paradigm for cGMP-directed therapy and prevention for colorectal tumors.


Coil embolization of a traumatic arteriovenous fistula of the lower extremity.
Arteriovenous fistulas (AVFs) are unusual connections between the arterial and venous system that bypass the normal anatomic capillary beds. This case report presents an arteriovenous fistula of the lower extremity caused by a penetrating injury. The patient was diagnosed by Duplex ultrasonography. Diagnostic catheterization of the affected limb showed a high-flow AVF between the left profunda femoris artery and left femoral vein with a deep femoral vein aneurysm measuring 9x4 cm. The AVF was successfully treated with coil embolization.


Felbamate but not phenytoin or gabapentin reduces glutamate release by blocking presynaptic NMDA receptors in the entorhinal cortex.
We have shown that a number of anticonvulsant drugs can reduce glutamate release at synapses in the rat entorhinal cortex (EC) in vitro. We have also shown that presynaptic NMDA receptors (NMDAr) tonically facilitate glutamate release at these synapses. In the present study we determined whether, phenytoin, gabapentin and felbamate may reduce glutamate release by blocking the presynaptic NMDAr. Whole cell patch clamp recordings of spontaneous excitatory postsynaptic currents (sEPSCs) were used as a monitor of presynaptic glutamate release. Postsynaptic NMDAr were blocked with internal dialysis with an NMDAr channel blocker. The antagonist, 2-AP5, reduced the frequency of sEPSCs by blocking the presynaptic facilitatory NMDAr, but did not occlude a reduction in sEPSC frequency by gabapentin or phenytoin. Felbamate also reduced sEPSC frequency, but this effect was occluded by prior application of 2-AP5. Thus, whilst all three drugs can reduce glutamate release, only the action of felbamate seems to be due to interaction with presynaptic NMDAr.


Piriformospora indica affects plant growth by auxin production.
Piriformospora indica has been shown to improve the growth of many plant species including Arabidopsis thaliana, but the mechanism by which this is achieved is still unclear. Arabidopsis root colonization by P. indica was examined in sterile culture on the medium of Murashige and Skoog. P. indica formed intracellular structures in Arabidopsis root epidermal cells and caused changes in root growth, leading to stunted and highly branched root systems. This effect was because of a diffusible factor and could be mimicked by IAA. In addition, P. indica was shown to produce IAA in liquid culture. We suggest that auxin production affecting root growth is responsible for, or at least contributes to, the beneficial effect of P. indica on its host plants.


Mesenchymal stem cells prime proliferating adult neural progenitors toward an oligodendrocyte fate.
Oligodendrogenesis encompasses lineage specification of neural progenitor cells (NPCs) and differentiation into oligodendrocytes that ultimately culminates in the myelination of central nervous system axons. Each individual process must be tightly regulated by extracellular and cell-intrinsic mechanisms, whose identities are barely understood. We had previously demonstrated that soluble factors derived from rat mesenchymal stem cells (MSCs) induce oligodendrogenesis in differentiating adult NPCs under differentiation conditions. However, since lineage specification predominantly occurs in proliferating progenitors and not necessarily during early differentiation, we investigated if soluble factors derived from MSCs are able to prime NPCs to the oligodendroglial fate already under proliferation conditions. Therefore, we analyzed the effects of a 3 weeks stimulation of adult NPCs under proliferation conditions with conditioned media derived from MSCs (MSC-CM) in terms of cell morphology, proliferation, cell-specific marker expression profile, response to growth factor withdrawal (GFW), cell-lineage restriction, and expression of glial fate determinants. While MSC-CM did not affect the proliferation rate of NPCs, it boosted the formation of 2', 3'-cyclic-nucleotide-3'-phosphodieesterase (CNPase)- and myelin basic protein-expressing oligodendrocytes after GFW, even when cells were exposed to an astrogenic milieu. Moreover, it reinforced the proper development of oligodendrocytes, since it ensured a sustained expression of the functional marker CNPase. Finally, the presence of MSC-CM reduced the anti-oligodendrogenic determinant Id2 in proliferating NPCs, thus increasing the relative proportion of the pro-oligodendrogenic factor Olig2 expression. In summary, MSCs prime proliferating progenitors and, thus, reinforce cell fate choice and accelerate differentiation toward the oligodendrocyte lineage. The present findings underscore the potential use of MSCs in cell therapies for remyelination such as in multiple sclerosis and spinal cord injury. Moreover, they urge the identification of the oligodendrogenic activity(ies) derived from MSCs to develop novel molecular therapies for demyelinating diseases.


Using transgenic reporters to visualize bone and cartilage signaling during development in vivo.
Green fluorescent protein was first used as a marker of protein expression in vivo 18 years ago, heralding the beginning of what became known as the Green Revolution. Since then, there has been an explosion in the number of transgenic lines in existence, and these transgenic tools are now being applied to skeletal research. Advances in transgenesis are also leading to increasing use of new model organisms for studying skeletogenesis. Such new models include the small teleosts zebrafish and medaka, which due to their optical translucency offer imaging possibilities in the live animals. In this review, we will introduce a number of recent advances in genetic engineering and transgenesis and the new genetic tools that are currently being developed. We will provide examples of how zebrafish and medaka transgenic lines are helping us to understand the behavior of skeletal cells in vivo. Finally, we will discuss future prospects for the application of transgenic technology to skeletal research.


Symmetric Peripheral Gangrene Associated with H1N1 Infection.
More and more cases of H1N1 influenza are being detected in India and so also the variety of complications this virus can cause. Here, we report a case of symmetric peripheral gangrene following H1N1 infection.


Importance of conversion for results obtained with laparoscopic colorectal surgery.
The need for a conversion is a problem inherent in laparoscopic surgery. The present study points up the significance of conversion for the results obtained with laparoscopic colorectal surgery and identifies the risk factors that establish the need for conversion. The study took the form of a multicentric, prospective, observational study within the Laparoscopic Colorectal Surgery Study Group. A total of 33 institutions in Germany, Austria, and Switzerland participated. The study period was 3.5 years. Cases were documented with the aid of a standardized questionnaire. Within the framework of the Laparoscopic Colorectal Surgery Study Group, a total of 1,658 patients were recruited to a multicenter study over a period of three and one-half years (from August 1, 1995 to February 1, 1999). The observed conversion rate was 5.2 percent (n = 86). The patients requiring a conversion were significantly heavier (body mass index, 26.5 vs. 24.9) than those undergoing pure laparoscopy. Resections of the rectum were associated with a higher risk for conversion (20.9 vs. 13 percent). Intraoperative complications occurred significantly more frequently in the conversion group (27.9 vs. 3.8 percent). The duration of the operation was significantly increased after conversion in a considerable proportion of the procedures performed. Postoperative morbidity (47.7 vs. 26.1 percent), mortality (3.5 vs. 1.5 percent), recovery time, and postoperative hospital stay were all negatively influenced by conversion, in part significantly. Institutions with experience of more than 100 laparoscopic colorectal procedures proved to have a significantly lower conversion rate than those with experience of fewer than 100 such interventions (4.3 vs. 6.9 percent). Although, of itself, conversion is not considered to be a complication of laparoscopic surgery, it is true that the postoperative course after conversion is associated with appreciably poorer results in terms of morbidity, mortality, convalescence, blood transfusion requirement, and postoperative hospital stay. The importance of experience in laparoscopic surgery can be demonstrated on the basis of the conversion rates. Careful patient selection oriented to the experience of the surgeon is required if we are to keep the conversion, morbidity, and mortality rates of laparoscopic colorectal procedures as low as possible.


Characterization of the Molecular Interactions That Govern the Packaging of Viral RNA Segments into Rift Valley Fever Phlebovirus Particles.
Rift Valley fever phlebovirus (RVFV) has a single-stranded, negative-sense RNA genome, consisting of L, M, and S segments. The virion carries two envelope glycoproteins, Gn and Gc, along with ribonucleoprotein complexes (RNPs), composed of encapsidated genomes carrying N protein and the viral polymerase, L protein. A quantitative analysis of the profile of viral RNA segments packaged into RVFV particles showed that all three genomic RNA segments had similar packaging abilities, whereas among antigenomic RNA segments, the antigenomic S RNA, which serves as the template for the transcription of mRNA expressing the RVFV virulence factor, NSs, displayed a significantly higher packaging ability. To delineate the factor(s) governing the packaging of RVFV RNA segments, we characterized the interactions between Gn and viral RNPs in RVFV-infected cells. Coimmunoprecipitation analysis demonstrated the interaction of Gn with N protein, L protein, and viral RNAs in RVFV-infected cells. Furthermore, UV-cross-linking and immunoprecipitation analysis revealed, for the first time in bunyaviruses, the presence of a direct interaction between Gn and all the viral RNA segments in RVFV-infected cells. Notably, analysis of the ability of Gn to bind to RVFV RNA segments indicated a positive correlation with their respective packaging abilities and highlighted a binding preference of Gn for antigenomic S RNA, among the antigenomic RNA segments, suggesting the presence of a selection mechanism for antigenomic S RNA incorporation into infectious RVFV particles. Collectively, the results of our study illuminate the importance of a direct interaction between Gn and viral RNA segments in determining their efficiency of incorporation into RVFV particles. IMPORTANCE Rift Valley fever phlebovirus, a bunyavirus, is a mosquito-borne, segmented RNA virus that can cause severe disease in humans and ruminants. An essential step in RVFV life cycle is the packaging of viral RNA segments to produce infectious virus particles for dissemination to new hosts. However, there are key gaps in knowledge regarding the mechanisms that regulate viral RNA packaging efficiency in bunyaviruses. Our studies investigating the mechanism of RNA packaging in RVFV revealed the presence of a direct interaction between the viral envelope glycoprotein, Gn, and the viral RNA segments in infected cells, for the first time in bunyaviruses. Furthermore, our data strongly indicate a critical role for the direct interaction between Gn and viral RNAs in determining the efficiency of incorporation of viral RNA segments into RVFV particles. Clarifying the fundamental mechanisms of RNA packaging in RVFV would be valuable for the development of antivirals and live-attenuated vaccines.


Toward molecular mechanism of xenon anesthesia: a link to studies of xenon complexes with small aromatic molecules.
The present study illustrates the steps toward understanding molecular mechanism of xenon anesthesia by focusing on a link to the structures and spectra of intermolecular complexes of xenon with small aromatic molecules. A primary cause of xenon anesthesia is attributed to inhibition of N-methyl-D-aspartate (NMDA) receptors by an unknown mechanism. Following the results of quantum mechanics/molecular mechanics (QM/MM) and molecular dynamics (MD) calculations we report plausible xenon action sites in the ligand binding domain of the NMDA receptor, which are due to interaction of xenon atoms with aromatic amino-acid residues. We rely in these calculations on computational protocols adjusted in combined experimental and theoretical studies of intermolecular complexes of xenon with phenol. Successful reproduction of vibrational shifts in molecular species upon complexation with xenon measured in low-temperature matrices allowed us to select a proper functional form in density functional theory (DFT) approach for use in QM subsystems, as well as to calibrate force field parameters for MD simulations. The results of molecular modeling show that xenon atoms can compete with agonists for a place in the corresponding protein cavity, thus indicating their active role in anesthetic action.


MUJA: manipulation under joint anesthesia/analgesia: a treatment approach for recalcitrant low back pain of synovial joint origin.
These four cases show how application of manipulation under joint anesthesia/analgesia (MUJA) may benefit the patient with low back pain (LBP) of synovial joint origin when prior treatment options fail. We propose that MUJA should be considered as a treatment option for those with recalcitrant synovial joint-mediated LBP. We report four cases of patients with LBP successfully treated by this protocol. These four patients were neurologically normal and suffered from lumbar zygapophyseal joint and/or sacroiliac joint mediated pain diagnosed by fluoroscopically guided, contrast enhanced, intra-articular anesthetic injections. There were various radiographic findings. All four individuals were treated with manipulation after facilitatory intra-articular anesthetic/corticosteroid injections into lumbar zygapophyseal and/or sacroiliac joints. Some also required physical therapy interventions to resolve their LBP. Anecdotal success with MUJA is enticing; MUJA appears to be a viable alternative for those patients in whom traditional treatment options for LBP of synovial joint origin have failed. However, prospective, quality research is required before MUJA can be formally validated or not in the treatment of documented lumbar zygapophyseal joint and/or sacroiliac joint mediated LBP.


Mirtazapine exerts astrocyte-mediated dopaminergic neuroprotection.
Mirtazapine, a noradrenergic and specific serotonergic antidepressant (NaSSA), is known to activate serotonin (5-HT) 1A receptor. Our recent study demonstrated that stimulation of astrocytic 5-HT1A receptors promoted astrocyte proliferation and upregulated antioxidative property in astrocytes to protect dopaminergic neurons against oxidative stress. Here, we evaluated the neuroprotective effects of mirtazapine against dopaminergic neurodegeneration in models of Parkinson's disease (PD). Mirtazapine administration attenuated the loss of dopaminergic neurons in the substantia nigra and increased the expression of the antioxidative molecule metallothionein (MT) in the striatal astrocytes of 6-hydroxydopamine (6-OHDA)-injected parkinsonian mice via 5-HT1A receptors. Mirtazapine protected dopaminergic neurons against 6-OHDA-induced neurotoxicity in mesencephalic neuron and striatal astrocyte cocultures, but not in enriched neuronal cultures. Mirtazapine-treated neuron-conditioned medium (Mir-NCM) induced astrocyte proliferation and upregulated MT expression via 5-HT1A receptors on astrocytes. Furthermore, treatment with medium from Mir-NCM-treated astrocytes protected dopaminergic neurons against 6-OHDA neurotoxicity, and these effects were attenuated by treatment with a MT-1/2-specific antibody or 5-HT1A antagonist. Our study suggests that mirtazapine could be an effective disease-modifying drug for PD and highlights that astrocytic 5-HT1A receptors may be a novel target for the treatment of PD.


Post-acute care reform: Implications and opportunities for hospitalists.
Nearly all practicing hospitalists have firsthand experience discharging patients to post-acute care (PAC), which is provided by inpatient rehabilitation facilities, skilled nursing facilities, or home healthcare providers. Many may not know that PAC is poised to undergo transformative change, spurred by recent legislation resulting in a range of reforms. These reforms have the potential to fundamentally reshape the relationship between hospitals and PAC providers. They have important implications for hospitalists and will open up opportunities for hospitalists to improve healthcare value. In this article, the authors explore the reasons for PAC reform and the scope of the reforms. Then they describe the implications for hospitalists and hospitalists' opportunities to Choose Wisely and improve healthcare value for the rapidly growing number of vulnerable older adults transitioning to PAC after hospital discharge.


Management of cervical deformity in ankylosing spondylitis.
Ankylosing spondylitis can lead to severe cervical kyphosis, causing problems with forward vision, swallowing, hygiene, patient functionality, and social outlook. Evaluation of patients with cervical flexion deformity includes assessment of global sagittal balance and chin-brow angle. The primary treatment in extreme disabling cases is surgical correction involving a posterior cervical extension osteotomy, which is a technically demanding procedure with considerable risk of neurological injury. To address the potential complications with extension osteotomy, the authors of several reports have described modifications to the surgical technique. These developments incorporate recent advances in anesthesia, neuromonitoring, and spinal instrumentation. Complications associated with the procedure include subluxation at the osteotomy level, spinal cord injury, radiculopathy, dysphagia, and pseudarthrosis. Although the risks of spinal correction are considerable, extension osteotomy remains an effective treatment modality for patients with disabling cervical flexion deformity.


Children recovered from stuttering without formal treatment: perceptual assessment of speech normalcy.
Current evidence suggests that young children who recover from stuttering are essentially stutter-free. However, there is no evidence to indicate if their speech is perceptually indistinguishable from normally fluent peers or whether they retain perceptually unusual speech. One important example of recovery from stuttering is children who have recovered without receiving formal treatment. An investigation was conducted to determine if the speech of these children is perceptually different from the speech of children who have never stuttered. Speakers consisted of 10 preschool and early school-age children documented as recovered from stuttering without benefit of formal treatment. In a series of studies they were compared with 10 children who had never stuttered. Three groups of judges-sophisticated, unsophisticated, and experienced-were separately asked, using videotaped speech samples of the children, to decide which samples were from children who used to stutter. Results revealed that the children who recovered from stuttering were perceptually indistinguishable from the normal controls. The same result was obtained regardless of whether the samples were presented in paired-stimulus or single-stimulus mode. Two of the groups of judges were also instructed to rate the speech naturalness of the speech samples. The speakers were not distinguished on this measure either. Methodological issues and the implications of the findings are discussed.


Axon terminals containing PACAP- and VIP-immunoreactivity form synapses with CRF-immunoreactive neurons in the dorsolateral division of the bed nucleus of the stria terminalis in the rat.
The bed nucleus of the stria terminalis (BST) is a highly heterogeneous forebrain structure, within which the median and lateral BST play distinct functional roles. The medial BST (BSTM) is thought to be related to sexual behavior, while the lateral BST (BSTL) may have a stress-related function. In the human brain, the BST shows marked sexual dimorphism in the distribution of vasoactive intestinal polypeptide (VIP) immunoreactive fibers and also contains a very high concentration of pituitary adenylate cyclase activating polypeptide (PACAP) immunoreactivity (ir). Using immunohistochemistry (IHC) to examine the rat brain, the present study found that both VIP and PACAP containing afferent fibers are abundant in the BSTLd (dorsolateral division of BST), but not in the BSTM. IHC did not reveal any apparent difference between the sexes in the size of distribution of either immunoreactivity. Double staining IHC showed that axonal terminals of both VIP and PACAP neurons were in close proximity to dendrites or perikarya of corticotropin releasing factor (CRF) neurons. At the electron microscopic level IHC revealed the presence of axodendritic or axosomatic synapses between VIP-ir and PACAP-ir axon terminals and CRF-ir neurons. Although the origin of PACAP-ir fibers in the BSTLd remains to be determined, these morphological findings suggest that PACAP and VIP regulate the activity of CRF neurons in the BSTLd as neurotransmitters or neuromodulators.


Iatrogenic lidocaine toxicosis in ewes.
Lidocaine intoxication developed inadvertently in a group of ewes. Six ewes used in a student laboratory session on laparotomy procedures were anesthetized locally with lidocaine hydrochloride (20 mg/kg) prior to simultaneous bilateral laparotomy. Shortly after the administration of lidocaine hydrochloride, the ewes developed muscle tremors and became recumbent, with a variety of neurologic signs including dullness, opisthotonos, odontoprisis, mydriasis, blindness, extensor rigidity, and convulsions. The ewes were given intravenous fluid therapy, and 90 minutes after the onset of signs, the ewes were standing, dull, and appetent. The laboratory session was rescheduled 2 weeks later, using the same ewes and a reduced dosage of lidocaine hydrochloride (10 mg/kg). There were no adverse effects and the simultaneous bilateral laparotomy procedure was completed without complications. The authors recommend the use of lidocaine hydrochloride at a dosage not exceeding 10 mg/kg for ewes requiring extensive local anesthesia.


Azathioprine and diffuse alveolar haemorrhage: the pharmacogenetics of thiopurine methyltransferase.
Current guidelines support the use of corticosteroids and azathioprine as one possible treatment strategy for idiopathic pulmonary fibrosis (IPF). However, some patients with genetic polymorphisms of thiopurine methyltransferase (TPMT) are at risk of severe azathioprine myelotoxicity. The current authors present the case of an 85-yr-old Caucasian male with IPF who developed diffuse alveolar haemorrhage as a complication of azathioprine-induced myelosuppression. Leukocyte genetic TPMT testing revealed that the patient had homozygous polymorphisms associated with the absence of TPMT activity and severe azathioprine-induced myelotoxicity. Thiopurine methyltransferase deficiency should be considered in patients who develop leukopenia early in treatment with azathiopurine, or who present with severe marrow suppression at usual doses. For centres with equipped laboratories, a dosing suggestion is provided based on thiopurine methyltransferase testing. Even with screening strategies, frequent monitoring of complete blood count and liver biochemistry should remain the mainstay of surveillance for azathioprine toxicity.


Neonatal epithelial hypoxia inducible factor-1α expression regulates the response of the lung to experimental asthma.
Allergic airway disease is characterized by a T helper type 2 cell-mediated airway inflammation and airway hyperresponsiveness. Little is known about the role of hypoxia-mediated signaling in the progression of the disease. To address this knowledge gap, a mouse model was created in which doxycycline exposure induces the functional deletion of hypoxia inducible factor-1α from alveolar type II and Clara cells of the lung. When hypoxia inducible factor-1α deletion was induced during the early postnatal development period of the lung, the mice displayed an enhanced response to the ovalbumin model of allergic airway disease. These hypoxia inducible factor-1α-deficient mice exhibit increased cellular infiltrates, eosinophilia in the lavage fluid and parenchyma, and T helper type 2 cytokines, as compared with ovalbumin-treated control mice. Moreover, these hypoxia inducible factor-1α-deficient mice display increased airway resistance when compared with their control counterparts. Interestingly, if the loss of hypoxia inducible factor-1α was induced in early adulthood, the exacerbated phenotype was not observed. Taken together, these results suggest that epithelial hypoxia inducible factor-1α plays an important role in establishing the innate immunity of the lung and epithelial-specific deficiency in the transcription factor, during early postnatal development, increases the severity of inflammation and functional airway resistance, following ovalbumin challenge. Finally, these results might explain some of the chronic respiratory pathology observed in premature infants, especially those that receive supplemental oxygen. This early hyperoxic exposure, from normal ambient and supplemental oxygen, would presumably inhibit normal hypoxia inducible factor-1α signaling, mimicking the functional deletion described.


Impact of Mistletoe Triterpene Acids on the Uptake of Mistletoe Lectin by Cultured Tumor Cells.
Complementary treatment possibilities for the therapy of cancer are increasing in demand due to the severe side effects of the standard cytostatics used in the first-line therapy. A common approach as a complementary treatment is the use of aqueous extracts of Viscum album L. (Santalaceace). The therapeutic activity of these extracts is attributed to Mistletoe lectins which are Ribosome-inactivating proteins type II. Besides these main constituents the extract of Viscum album L. comprises also a mixture of lipophilic ingredients like triterpene acids of the oleanane, lupane and ursane type. However, these constituents are not contained in commercially available aqueous extracts due to their high lipophilicity and insolubility in aqueous extraction media. To understand the impact of the extract ingredients in cancer therapy, the intracellular uptake of the mistletoe lectin I (ML) by cultured tumor cells was investigated in relation to the mistletoe triterpene acids, mainly oleanolic acid. Firstly, these hydrophobic triterpene acids were solubilized using cyclodextrins ("TT" extract). Afterwards, the uptake of either single compounds (isolated ML and the aqueous "viscum" extract) or in combination with the TT extract (ML+TT, viscumTT), was analyzed. The uptake of ML was studied inTHP-1-, HL-60-, 143B- and Ewing TC-71-cells and determined after 30, 60 and 120 minutes by an enzyme linked immunosorbent assay which quantifies the A-chain of the hololectin. It could be shown that the intracellular uptake after 120 minutes amounted to 20% in all cell lines after incubation with viscumTT. The studies further revealed that the uptake in THP-1-, HL-60- and Ewing TC-71-cells was independent of the addition of TT extract. Interestingly, the uptake of ML by 143B-cells could only be measured after addition of triterpenes pointing to resistance to mistletoe lectin.


Tiletamine and zolazepam for immobilization of wild lions and leopards.
A 1:1 mixture of tiletamine and zolazepam was used for the immobilization of lions on 26 occasions and of leopards on 22 occasions. There was a significant (P less than 0.001) positive linear relationship between duration of anesthesia and dosage for all animals in which the duration was recorded (n = 36). This response could be divided into 4 separate regressions according to species and sex; lions being more susceptible to the drug than leopards and males more so than females. When dosage was expressed in terms of metabolic weight, the duration of anesthesia depended on the sex of the animal rather than the species, males being anesthetized 15 minutes longer than females for a given dosage. The threshold dosage was higher in leopards than in lions. The amount of time that could be spent working on the animal immobilized by the drug, compared with the total time committed to its induction, anesthesia, and recovery was 35 to 55%, a proportion that is similar to that associated with the use of other irreversible intramuscular anesthetics.


State-dependent changes of extracellular glutamate in the medial preoptic area in freely behaving rats.
Microdialysis technique was applied to the medial preoptic area (mPOA) of the hypothalamus, a crucial site for the regulation of sleep, in order to analyze the interrelationship between the extracellular level of glutamate (Glu) and the vigilance states. Dialysates from the mPOA were sampled at 5-min intervals for 2-h diurnal period with a perfusion rate of 2.0 microliters/min in freely moving rats, whose sleep-waking behaviors were polysomnographically monitored. Extracellular Glu increased during wakefulness, exhibiting a peak at the transition period from wakefulness to non-rapid-eye-movement sleep (NREMS) (18.0% above the average), whereas it decreased during NREMS (9.2% below the average). It is likely that Glu in the mPOA is dynamically involved in the alterations of the vigilance states.


Non-coding genomic regions possessing enhancer and silencer potential are associated with healthy aging and exceptional survival.
We have completed a genome-wide linkage scan for healthy aging using data collected from a family study, followed by fine-mapping by association in a separate population, the first such attempt reported. The family cohort consisted of parents of age 90 or above and their children ranging in age from 50 to 80. As a quantitative measure of healthy aging, we used a frailty index, called FI34, based on 34 health and function variables. The linkage scan found a single significant linkage peak on chromosome 12. Using an independent cohort of unrelated nonagenarians, we carried out a fine-scale association mapping of the region suggestive of linkage and identified three sites associated with healthy aging. These healthy-aging sites (HASs) are located in intergenic regions at 12q13-14. HAS-1 has been previously associated with multiple diseases, and an enhancer was recently mapped and experimentally validated within the site. HAS-2 is a previously uncharacterized site possessing genomic features suggestive of enhancer activity. HAS-3 contains features associated with Polycomb repression. The HASs also contain variants associated with exceptional longevity, based on a separate analysis. Our results provide insight into functional genomic networks involving non-coding regulatory elements that are involved in healthy aging and longevity.


Interfacial Photoelectrochemical Catalysis: Solar-Induced Green Synthesis of Organic Molecules.
Many oxidation and reduction reactions in conventional organic synthesis rely on harsh conditions, toxic or corrosive substances, and environmentally damaging chemicals. In addition, competing reactions may take place, some of which produce hazardous waste products and, therefore, reaction selectivity suffers. To overcome such synthetic drawbacks, an enormous effort is being devoted to find alternative processes that operate much more efficiently, requiring milder conditions to contribute to a greener economy and provide urgently needed new pathways with enhanced selectivity. Fortunately, there is a strategy that has attracted global interest from multiple disciplines that involves the use of sunlight to perform artificial photosynthesis, in which a photoelectrochemical cell splits water into hydrogen fuel, reduces CO2 into "solar" fuels, and more recently, convert organic chemicals into higher value products. Recently, photoanode and photocathode materials have emerged as useful tools to perform organic oxidations and reductions for the synthesis of important molecules, other than just hydrogen or oxygen. Whereas many studies have focused on the degradation of unwanted and dangerous chemicals, solar-induced organic transformations have attracted much less attention. This Minireview summarizes some of latest research efforts in using photoelectrochemical cells to facilitate organic oxidation and reduction reactions to avoid valuable substances while avoiding toxic reagents and expensive precious metal catalysts. Future developments that will enable such technologies to broaden their scope are also considered.


Intraspecific divergence of ionoregulatory physiology in the euryhaline teleost Fundulus heteroclitus: possible mechanisms of freshwater adaptation.
We examined intraspecific variation in ionoregulatory physiology within euryhaline killifish, Fundulus heteroclitus, to understand possible mechanisms of freshwater adaptation in fish. Pronounced differences in freshwater tolerance existed between northern (2% mortality) and southern (19% mortality) killifish populations after transfer from brackish water (10 g l(-1)) to freshwater. Differences in Na(+) regulation between each population might partially account for this difference in tolerance, because plasma Na(+) was decreased for a longer period in southern survivors than in northerns. Furthermore, northern fish increased Na(+)/K(+)-ATPase mRNA expression and activity in their gills to a greater extent 1-14 days after transfer than did southerns, which preceded higher whole-body net flux and unidirectional influx of Na(+) at 14 days. All observed differences in Na(+) regulation were small, however, and probably cannot account for the large differences in mortality. Differences in Cl(-) regulation also existed between populations. Plasma Cl(-) was maintained in northern fish, but in southerns, plasma Cl(-) decreased rapidly and remained low for the duration of the experiment. Correspondingly, net Cl(-) loss from southern fish remained high after transfer, while northerns eliminated Cl(-) loss altogether. Elevated Cl(-) loss from southern fish in freshwater was possibly due to a persistence of seawater gill morphology, as paracellular permeability (indicated by extrarenal clearance rate of PEG-4000) and apical crypt density in the gills (detected using scanning electron microscopy) were both higher than in northern fish. These large differences in the regulation of Cl(-) balance probably contributed to the marked differences in mortality after freshwater transfer. Glomerular filtration rate and urination frequency were also lower in southerns. Taken together, these data suggest that northern killifish are better adapted to freshwater environments and that minimizing Cl(-) imbalance appears to be the key physiological difference accounting for their greater freshwater tolerance.


Phylogenetically diverse Bradyrhizobium genospecies nodulate Bambara groundnut (Vigna subterranea L. Verdc) and soybean (Glycine max L. Merril) in the northern savanna zones of Ghana.
A total of 102 bacterial strains isolated from nodules of three Bambara groundnut and one soybean cultivars grown in nineteen soil samples collected from northern Ghana were characterized using multilocus gene sequence analysis. Based on a concatenated sequence analysis (glnII-rpoB-recA-gyrB-atpD-dnaK), 54 representative strains were distributed in 12 distinct lineages, many of which were placed mainly in the Bradyrhizobium japonicum and Bradyrhizobium elkanii supergroups. Twenty-four of the 54 representative strains belonged to seven putative novel species, while 30 were conspecific with four recognized Bradyrhizobium species. The nodA phylogeny placed all the representative strains in the cosmopolitan nodA clade III. The strains were further separated in seven nodA subclusters with reference strains mainly of African origin. The nifH phylogeny was somewhat congruent with the nodA phylogeny, but both symbiotic genes were mostly incongruent with the core housekeeping gene phylogeny indicating that the strains acquired their symbiotic genes horizontally from distantly related Bradyrhizobium species. Using redundancy analysis, the distribution of genospecies was found to be influenced by the edaphic factors of the respective sampling sites. In general, these results mainly underscore the high genetic diversity of Bambara groundnut-nodulating bradyrhizobia in Ghanaian soils and suggest a possible vast resource of adapted inoculant strains.


Genetic analysis of the major gene plus polygene model in soybean resistance to Leguminivora glycinivorella.
In order to investigate the genetic characteristics of soybean Leguminivora glycinivorella resistance and to improve soybean resistance insectivorous breeding efficiency by applying the multi-generation joint analysis method of the major gene plus polygene model, 5 pedigrees and generations (P1, F1, P2, F2, and F2:3) were used as the materials to perform the soybean L. glycinivorella resistance multi-generation joint analysis. The results showed that soybean resistance to L. glycinivorella was controlled and inherited by an additive major gene plus additive, dominant polygene. The major gene had a negative additive effect (d = -0.1633). The combination of the anti-L. glycinivorella genes showed negative heterosis. Because the polygene additive effects were positive, the polygene effects would increase the insect herbivory rate in the F1 generation. This hybrid combination showed an insect herbivory rate polygenic heritability of 21.9556 and 54.3490% in the F2 and F2:3 pedigrees, which presented a high heritability. Therefore, it was appropriate to perform the selective breeding of the insect herbivory rate in the late generation.


Discovery of Highly Efficient Novel Antifungal Lead Compounds Targeting Succinate Dehydrogenase: Pyrazole-4-carboxamide Derivatives with an N-Phenyl Substituted Amide Fragment.
Developing environmentally friendly fungicides is crucial to tackle the issue of rising pesticide resistance. In this study, a series of novel pyrazole-4-carboxamide derivatives containing N-phenyl substituted amide fragments were designed and synthesized. The structures of target compounds were confirmed by 1H NMR, 13C NMR, and HRMS, and the crystal structure of the most active compound N-(1-(4-(4-(tert-butyl)benzamido)phenyl)propan-2-yl)-3-(difluoromethyl)-N-methoxy-1-methyl-1H-pyrazole-4-carboxamide (U22) was further determined by X-ray single-crystal diffraction. The bioassay results indicated that the 26 target compounds possessed good in vitro antifungal activity against Sclerotinia sclerotiorum with EC50 values for compounds U12, U13, U15, U16, U18, U22, and U23 being 4.17 ± 0.46, 8.04 ± 0.71, 7.01 ± 0.71, 12.77 ± 1.00, 8.11 ± 0.70, 0.94 ± 0.11, and 9.48 ± 0.83 μg·mL-1, respectively, which were the similar to controls bixafen (6.70 ± 0.47 μg·mL-1), fluxapyroxad (0.71 ± 0.14 μg·mL-1), and pydiflumetofen (0.06 ± 0.01 μg·mL-1). Furthermore, in vivo antifungal activity results against S. sclerotiorum indicated that compounds U12 (80.6%) and U22 (89.9%) possessed excellent preventative efficacy at 200 μg·mL-1, which was the same as the control pydiflumetofen (82.4%). Scanning electron microscopy and transmission electron microscopy studies found that the compound U22 could destroy the hyphal morphology and damage mitochondria, cell membranes, and vacuoles. The results of molecular docking of compound U22 and pydiflumetofen with succinate dehydrogenase (SDH) indicated they interact well with the active site of SDH. This study validated our approach and design strategy to produce compounds with an enhanced biological activity as compared to the parent structure.


Corneal Neovascularization: A Combined Approach of Bevacizumab and Suramin Showed Increased Antiangiogenic Effect Through Downregulation of BFGF and P2X2.
The objective is to analyze the antiangiogenic mechanism of suramab, a pharmaceutical compound of bevacizumab and suramin, in a rabbit model of corneal angiogenesis. Corneal neovascularization was induced in four groups of six New Zealand White rabbits by applying a filter paper disk soaked in 1 M Na (OH) on the central cornea. Group one was treated after injury with intravenous suramab at a dose equivalent to 3 mg/kg of bevacizumab and 10 mg/kg of suramin. Group two was treated with intravenous bevacizumab (5 mg/kg). Group three was treated with 10 mg/kg of suramin while the control group received no treatment. Digital photographs were taken at days 9, 15, 21, and 35. Neovessel formation was quantified giving a 0-4 score to each quadrant according to the centripetal growth of the longest vessel (neovessel index, NVI). Animals were sacrificed at day 35. Corneas were processed for histology, immunohistochemistry, and Western-blot using primary antibodies against P2X2, basic fibroblast growth factor (bFGF), LYVE-1, PECAM-1, and vascular endothelial growth factor-A (VEGF-A). Suramab significantly reduced neovessel growth (mean NVI: 4.2) compared to bevacizumab (8.4), suramin (7.22), and control animals (12.2) at 35 days post-injury (p < 0.01). A lower protein expression of P2X2, bFGF, LYVE-1, PECAM-1, and VEGF-A was found in the cornea of suramab animals than in the other groups of animals. Joint downregulation of bFGF, P2X2, bFGF, and LYVE-1 constitutes a mechanism that induces greater and longer inhibition of corneal angiogenesis. Results might be relevant to ophthalmic care. Ocular administration of suramab is currently being investigated.


Biochemical properties of alcohol dehydrogenase from Drosophila lebanonensis.
Purified Drosophila lebanonensis alcohol dehydrogenase (Adh) revealed one enzymically active zone in starch gel electrophoresis at pH 8.5. This zone was located on the cathode side of the origin. Incubation of D. lebanonensis Adh with NAD+ and acetone altered the electrophoretic pattern to more anodal migrating zones. D. lebanonensis Adh has an Mr of 56,000, a subunit of Mr of 28 000 and is a dimer with two active sites per enzyme molecule. This agrees with a polypeptide chain of 247 residues. Metal analysis by plasma emission spectroscopy indicated that this insect alcohol dehydrogenase is not a metalloenzyme. In studies of the substrate specificity and stereospecificity, D. lebanonensis Adh was more active with secondary than with primary alcohols. Both alkyl groups in the secondary alcohols interacted hydrophobically with the alcohol binding region of the active site. The catalytic centre activity for propan-2-ol was 7.4 s-1 and the maximum velocity of most secondary alcohols was approximately the same and indicative of rate-limiting enzyme-coenzyme dissociation. For primary alcohols the maximum velocity varied and was much lower than for secondary alcohols. The catalytic centre activity for ethanol was 2.4 s-1. With [2H6]ethanol a primary kinetic 2H isotope effect of 2.8 indicated that the interconversion of the ternary complexes was rate-limiting. Pyrazole was an ethanol-competitive inhibitor of the enzyme. The difference spectra of the enzyme-NAD+-pyrazole complex gave an absorption peak at 305 nm with epsilon 305 14.5 X 10(3) M-1 X cm-1. Concentrations and amounts of active enzyme can thus be determined. A kinetic rate assay to determine the concentration of enzyme active sites is also presented. This has been developed from active site concentrations established by titration at 305 nm of the enzyme and pyrazole with NAD+. In contrast with the amino acid composition, which indicated that D. lebanonensis Adh and the D. melanogaster alleloenzymes were not closely related, the enzymological studies showed that their active sites were similar although differing markedly from those of zinc alcohol dehydrogenases.


Architecture of the sperm whale forehead facilitates ramming combat.
Herman Melville's novel Moby Dick was inspired by historical instances in which large sperm whales (Physeter macrocephalus L.) sank 19th century whaling ships by ramming them with their foreheads. The immense forehead of sperm whales is possibly the largest, and one of the strangest, anatomical structures in the animal kingdom. It contains two large oil-filled compartments, known as the "spermaceti organ" and "junk," that constitute up to one-quarter of body mass and extend one-third of the total length of the whale. Recognized as playing an important role in echolocation, previous studies have also attributed the complex structural configuration of the spermaceti organ and junk to acoustic sexual selection, acoustic prey debilitation, buoyancy control, and aggressive ramming. Of these additional suggested functions, ramming remains the most controversial, and the potential mechanical roles of the structural components of the spermaceti organ and junk in ramming remain untested. Here we explore the aggressive ramming hypothesis using a novel combination of structural engineering principles and probabilistic simulation to determine if the unique structure of the junk significantly reduces stress in the skull during quasi-static impact. Our analyses indicate that the connective tissue partitions in the junk reduce von Mises stresses across the skull and that the load-redistribution functionality of the former is insensitive to moderate variation in tissue material parameters, the thickness of the partitions, and variations in the location and angle of the applied load. Absence of the connective tissue partitions increases skull stresses, particularly in the rostral aspect of the upper jaw, further hinting of the important role the architecture of the junk may play in ramming events. Our study also found that impact loads on the spermaceti organ generate lower skull stresses than an impact on the junk. Nevertheless, whilst an impact on the spermaceti organ would reduce skull stresses, it would also cause high compressive stresses on the anterior aspect of the organ and the connective tissue case, possibly making these structures more prone to failure. This outcome, coupled with the facts that the spermaceti organ houses sensitive and essential sonar producing structures and the rostral portion of junk, rather than the spermaceti organ, is frequently a site of significant scarring in mature males suggest that whales avoid impact with the spermaceti organ. Although the unique structure of the junk certainly serves multiple functions, our results are consistent with the hypothesis that the structure also evolved to function as a massive battering ram during male-male competition.


Safety and efficacy of e-cigarettes in those with atherosclerotic disease: a review.
Smoking cessation is the most effective intervention to reduce mortality in patients with established atherosclerotic cardiovascular disease (ASCVD), with 'e-cigarettes' becoming an increasingly used intervention to achieve smoking cessation. The current review aims to summarise the current evidence base for their efficacy and safety in the ASCVD cohort. A search of the PUBMED and MEDLINE databases using the terms 'e-cigarette', 'cessation', 'safety' and 'efficacy' since 2012 yielded 706 results. Both observational and experimental studies were included, while those with an unavailable full text, non-English or duplicates were excluded, yielding 78 relevant articles, with 13 subsequent additional articles included from a search of reference lists, for a total of 91 included papers. E-cigarette vapour contains many known pro-atherosclerotic substances and has been demonstrated to potentiate traditional atherosclerotic mechanisms. While e-cigarettes may be more effective in promoting smoking cessation in the general population over a medium term (>6 months), when compared with nicotine replacement therapy (NRT), few studies specifically examined those with ASCVD, despite the latter having a higher baseline quit rate (52% vs 2%). Most studies compare e-cigarettes with NRT alone and do not include pharmacotherapy, which may be more effective in the ASCVD cohort. The single randomised controlled trial addressing the research question favoured traditional methods. Those that successfully quit smoking using e-cigarettes are more likely to continue to use the intervention at 1 year (90% vs 9%). Conflicting advice exists regarding the utilisation of e-cigarettes for smoking cessation. E-cigarettes may be inferior to standard care for smoking cessation in those with ASCVD, and their use is likely to promote the key drivers of the atherosclerotic process already active in this cohort.


RNA-DNA Hybrids Support Recombination-Based Telomere Maintenance in Fission Yeast.
A subset of cancers rely on telomerase-independent mechanisms to maintain their chromosome ends. The predominant "alternative lengthening of telomeres" pathway appears dependent on homology-directed repair (HDR) to maintain telomeric DNA. However, the molecular changes needed for cells to productively engage in telomeric HDR are poorly understood. To gain new insights into this transition, we monitored the state of telomeres during serial culture of fission yeast (Schizosaccharomyces pombe) lacking the telomerase recruitment factor Ccq1. Rad52 is loaded onto critically short telomeres shortly after germination despite continued telomere erosion, suggesting that recruitment of recombination factors is not sufficient to maintain telomeres in the absence of telomerase function. Instead, survivor formation coincides with the derepression of telomeric repeat-containing RNA (TERRA). In this context, degradation of TERRA associated with the telomere in the form of R-loops drives a severe growth crisis, ultimately leading to a novel type of survivor with linear chromosomes and altered cytological telomere characteristics, including the loss of the shelterin component Rap1 (but not the TRF1/TRF2 ortholog, Taz1) from the telomere. We demonstrate that deletion of Rap1 is protective in this context, preventing the growth crisis that is otherwise triggered by degradation of telomeric R-loops in survivors with linear chromosomes. These findings suggest that upregulation of telomere-engaged TERRA, or altered recruitment of shelterin components, can support telomerase-independent telomere maintenance.


ZapA, a virulence factor in a rat model of Proteus mirabilis-induced acute and chronic prostatitis.
Our knowledge of pathogenesis has benefited from a better understanding of the roles of specific virulence factors in disease. To determine the role of the virulence factor ZapA, a 54-kDa metalloproteinase of Proteus mirabilis, in prostatitis, rats were infected with either wild-type (WT) P. mirabilis or its isogenic ZapA(-) mutant KW360. The WT produced both acute and chronic prostatitis showing the typical histological progressions that are the hallmarks of these diseases. Infection with the ZapA(-) mutant, however, resulted in reduced levels of acute prostatitis, as determined from lower levels of tissue damage, bacterial colonization, and inflammation. Further, the ZapA(-) mutant failed to establish a chronic infection, in that bacteria were cleared from the prostate, inflammation was resolved, and tissue was seen to be healing. Clearance from the prostate was not the result of a reduced capacity of the ZapA(-) mutant to form biofilms in vitro. These finding clearly define ZapA as an important virulence factor in both acute and chronic bacterial prostatitis.