Cutaneous mucinosis in shar-pei dogs is due to hyaluronic acid deposition and is associated with high levels of hyaluronic acid in serum.
Cutaneous mucinosis affects primarily shar-pei dogs. Hyaluronic acid (HA) is considered to be the main component of mucin and CD44 is the major cell surface receptor of HA, necessary for its uptake and catabolism. The aims of this study were to identify the composition of the mucin in cutaneous mucinosis of shar-pei dogs, investigate the correlation between the deposition of HA and the expression of CD44, and determine whether shar-pei dogs with cutaneous mucinosis presented with elevated levels of serum HA. In skin biopsies, the mucinous material was stained intensely with Alcian blue and bound strongly by the hyaluronan-binding protein. No correlation was found between the degree of HA deposition in the dermis and the expression of CD44 in the skin of shar-pei dogs affected or unaffected by cutaneous mucinosis. A clear positive correlation was found between the existence of clinical mucinosis and the serum HA concentration. In control dogs, serum HA ranged from 155.53 to 301.62 microg L(-1) in shar-pei dogs; without mucinosis it ranged from 106.72 to 1251.76 microg L(-1) and in shar-pei dogs with severe mucinosis it ranged between 843.51 to 2330.03 microg L(-1). Altogether, the results reported here suggest that mucinosis of shar-pei dogs is probably the consequence of a genetic defect in the metabolism of HA.


Interpretation of the Seattle Angina Questionnaire as an Outcome Measure in Clinical Trials and Clinical Care: A Review.
Patient-reported outcomes are increasingly used as end points in clinical trials, assessments in clinical care, and tools for population health, with an increasing role in quality assessment. For patients with coronary artery disease, the Seattle Angina Questionnaire (SAQ) has emerged as the most commonly used measure of disease-specific health status to quantify patients' symptoms of angina and the extent to which their angina affects their functioning and quality of life. This review explains how to interpret the SAQ and describes the construction and face validity of the SAQ, focusing on aligning scores and changes in scores with clinical constructs. The SAQ asks questions similar to those an experienced clinician would ask of a patient with stable ischemic heart disease. Therefore, SAQ scores can be aligned with clinical constructs (eg, scores on the SAQ angina frequency scale of 0-30 points indicate daily angina, 31-60 points indicate weekly angina, 61-99 points indicate monthly angina, and 100 points indicate no angina), and changes in scores can be described by aligning them with changes in question responses. After clinical thresholds are defined, it is important for clinical trials to not simply report mean differences between treatment arms but to also report the distributions of patients who have had clinically important benefits so that a number needed to treat can be generated. The widespread use of the SAQ is a consequence of its well-established validity, reproducibility, prognostic importance, and sensitivity to clinical change. Nevertheless, interpreting the SAQ can be challenging because of lack of familiarity with the clinical importance of its domains, either cross-sectionally or longitudinally. This review provides an overview of the interpretability of the SAQ as a foundation for its use as an end point in clinical trials, a tool to support more patient-centered care, and a means of facilitating population health strategies to provide a better foundation for the integration of patient experiences with clinical care.


Myostatin knockout drives browning of white adipose tissue through activating the AMPK-PGC1α-Fndc5 pathway in muscle.
Myostatin (Mstn) is predominantly expressed in skeletal muscles and plays important roles in regulating muscle growth and development, as well as fat deposition. Mstn-knockout (Mstn(-/-)) mice exhibit increased muscle mass due to both hypertrophy and hyperplasia, and leaner body composition due to reduced fat mass. Here, we show that white adipose tissue (WAT) of Mstn(-/-) develops characteristics of brown adipose tissue (BAT) with dramatically increased expression of BAT signature genes, including Ucp1 and Pgc1α, and beige adipocyte markers Tmem26 and CD137. Strikingly, the observed browning phenotype is non-cell autonomous and is instead driven by the newly defined myokine irisin (Fndc5) secreted from Mstn(-/-) skeletal muscle. Within the muscle, Mstn(-/-) leads to increased expression of AMPK and its phosphorylation, which subsequently activates PGC1α and Fndc5. Together, our study defines a paradigm of muscle-fat crosstalk mediated by Fndc5, which is up-regulated and secreted from muscle to induce beige cell markers and the browning of WAT in Mstn(-/-) mice. These results suggest that targeting muscle Mstn and its downstream signaling represents a therapeutic approach to treat obesity and type 2 diabetes.


Unchanged density of caudate nucleus dopamine uptake sites in depressed suicide victims.
In depressive states, theories concerning serotonin and norepinephrine have been dominating, but there are several lines of evidence indicating the involvement of the dopamine system as well, especially in suicidal depression. In this post-mortem study, the binding of the ligand [3H]WIN 35,428 to dopamine uptake sites in the caudate nucleus was investigated in 13 depressed suicide victims and 19 controls. There were no differences in Bmax or Kd between the suicide group and controls. Subdividing the suicide group into subgroups regarding the presence of major depression, antidepressant medication and suicide method, respectively, did not yield any differences. Previous findings regarding reduced CSF HVA in suicidal depression and indications of striatal dopaminergic biochemical and receptor changes in depression seem, according to the present study, not to be reflected by alterations in density or affinity of dopamine uptake sites in depressed suicide victims.


Crystal structure and kinetic analysis of the class B3 di-zinc metallo-β-lactamase LRA-12 from an Alaskan soil metagenome.
We analyzed the kinetic properties of the metagenomic class B3 β-lactamase LRA-12, and determined its crystallographic structure in order to compare it with prevalent metallo-β-lactamases (MBLs) associated with clinical pathogens. We showed that LRA-12 confers extended-spectrum resistance on E. coli when expressed from recombinant clones, and the MIC values for carbapenems were similar to those observed in enterobacteria expressing plasmid-borne MBLs such as VIM, IMP or NDM. This was in agreement with the strong carbapenemase activity displayed by LRA-12, similar to GOB β-lactamases. Among the chelating agents evaluated, dipicolinic acid inhibited the enzyme more strongly than EDTA, which required pre-incubation with the enzyme to achieve measurable inhibition. Structurally, LRA-12 contains the conserved main structural features of di-zinc class B β-lactamases, and presents unique structural signatures that differentiate this enzyme from others within the family: (i) two loops (α3-β7 and β11-α5) that could influence antibiotic entrance and remodeling of the active site cavity; (ii) a voluminous catalytic cavity probably responsible for the high hydrolytic efficiency of the enzyme; (iii) the absence of disulfide bridges; (iv) a unique Gln116 at metal-binding site 1; (v) a methionine residue at position 221that replaces Cys/Ser found in other B3 β-lactamases in a predominantly hydrophobic environment, likely playing a role in protein stability. The structure of LRA-12 indicates that MBLs exist in wild microbial populations in extreme environments, or environments with low anthropic impact, and under the appropriate antibiotic selective pressure could be captured and disseminated to pathogens.


Developing Self-Assembled Starch Nanoparticles in Starch Nanocomposite Films.
Starch nanoparticles (SNPs) are synthesized by different precipitation techniques using corn starch, and SNP films are prepared by the evaporation casting method. The morphological study is investigated by scanning electron microscopy (SEM) and atomic force microscopy (AFM). The distribution and sizes of precipitated SNPs after synthesizing are discovered by these methods as well. The crystallinity of the SNPs is studied by the X-ray diffractometry (XRD) method that demonstrates reduction compared to neat starch granules, and it is changed from A-style to VH-style after precipitation. The chemical bonding of different SNPs after the nanoprecipitation is analyzed by Fourier transform infrared spectroscopy (FT-IR). Thermogravimetric analysis (TGA) demonstrates the decomposition of starch nanoparticles and the starch matrix that is related to the depolymerization of carbon chains in the range of 260 to 350 °C. The mechanical properties of the SNP films versus the temperature changing are discovered by dynamic mechanical analysis (DMA). The water contact angles of SNP films are measured using a goniometer, and the results showed the hydrophobic surfaces of the prepared films. Our study indicates that SNPs have a promising impact on the properties of corn starch films, which would be useful in biodegradable packaging material.


RNA interference pathways in filamentous fungi.
RNA interference is a conserved homology-dependent post-transcriptional/transcriptional gene silencing mechanism in eukaryotes. The filamentous fungus Neurospora crassa is one of the first organisms used for RNAi studies. Quelling and meiotic silencing by unpaired DNA are two RNAi-related phenomena discovered in Neurospora, and their characterizations have contributed significantly to our understanding of RNAi mechanisms in eukaryotes. A type of DNA damage-induced small RNA, microRNA-like small RNAs and Dicer-independent small silencing RNAs were recently discovered in Neurospora. In addition, there are at least six different pathways responsible for the production of these small RNAs, establishing this fungus as an important model system to study small RNA function and biogenesis. The studies in Cryphonectria, Mucor, Aspergillus and other species indicate that RNAi is widely conserved in filamentous fungi and plays important roles in genome defense. This review summarizes our current understanding of RNAi pathways in filamentous fungi.


Episodic and chronic migraine headache: breaking down barriers to optimal treatment and prevention.
Migraine is a common disabling primary headache disorder that affects an estimated 36 million Americans. Migraine headaches often occur over many years or over an individual's lifetime. By definition, episodic migraine is characterized by headaches that occur on fewer than 15 days per month. According to the recent International Classification of Headache Disorders (third revision) beta diagnostic criteria, chronic migraine is defined as "headaches on at least 15 days per month for at least 3 months, with the features of migraine on at least 8 days per month." However, diagnostic criteria distinguishing episodic from chronic migraine continue to evolve. Persons with episodic migraine can remit, not change, or progress to high-frequency episodic or chronic migraine over time. Chronic migraine is associated with a substantially greater personal and societal burden, more frequent comorbidities, and possibly with persistent and progressive brain abnormalities. Many patients are poorly responsive to, or noncompliant with, conventional preventive therapies. The primary goals of migraine treatment include relieving pain, restoring function, and reducing headache frequency; an additional goal may be preventing progression to chronic migraine. Although all migraineurs require abortive treatment, and all patients with chronic migraine require preventive treatment, there are no definitive guidelines delineating which persons with episodic migraine would benefit from preventive therapy. Five US Food and Drug Association strategies are approved for preventing episodic migraine, but only injections with onabotulinumtoxinA are approved for preventing chronic migraine. Identifying persons who require migraine prophylaxis and selecting and initiating the most appropriate treatment strategy may prevent progression from episodic to chronic migraine and alleviate the pain and suffering associated with frequent migraine.


The influence of hydrogen peroxide and hydrogen on the corrosion of simulated spent nuclear fuel.
The synergistic influence between H(2)O(2) and H(2) on the corrosion of SIMFUEL (simulated spent nuclear fuel) has been studied in solutions with and without added HCO(3)(-)/CO(3)(2-). The response of the surface to increasing concentrations of added H(2)O(2) was monitored by measuring the corrosion potential in either Ar or Ar/H(2)-purged solutions. Using X-ray photoelectron spectroscopy it was shown that the extent of surface oxidation (U(V) + U(VI) content) was directly related to the corrosion potential. Variations in corrosion potential with time, redox conditions, HCO(3)(-)/CO(3)(2-) concentration, and convective conditions showed that surface oxidation induced by H(2)O(2) could be reversed by reaction with H(2), the latter reaction occurring dominantly on the noble metal particles in the SIMFUEL. For sufficiently large H(2)O(2) concentrations, the influence of H(2) was overwhelmed and irreversible oxidation of the surface to U(VI) occurred. Subsequently, corrosion was controlled by the chemical dissolution rate of this U(VI) layer.


Activation of adenosine receptor A2A increases HSC proliferation and inhibits death and senescence by down-regulation of p53 and Rb.
During fibrosis hepatic stellate cells (HSC) undergo activation, proliferation, and senescence but the regulation of these important processes is poorly understood. The adenosine A2A receptor (A2A) is known to be present on HSC, and its activation results in liver fibrosis. In this study, we tested if A2A has a role in the regulation of HSC proliferation, apoptosis, senescence, and the relevant molecular mechanism. The ability of adenosine to regulate p53 and Rb protein levels, proliferation, apoptosis and senescence was tested in the human HSC cell line LX-2 and rat primary HSC. Adenosine receptor activation down-regulates p53 and Rb protein levels, increases BrdU incorporation and increases cell survival in LX-2 cells and in primary rat HSC. These effects of NECA were reproduced by an adenosine A2A receptor specific agonist (CGS21680) and blocked by a specific antagonist (ZM241385). By day twenty-one of culture primary rat HSC entered senescence and expressed β-gal which was significantly inhibited by NECA. Furthermore, NECA induced down regulation of p53 and Rb and Rac1, and decreased phosphorylation of p44-42 MAP Kinase in LX-2 cells and primary rat HSC. These effects were reproduced by the cAMP analog 8-Bromo-cAMP, and the adenylyl cyclase activator forskolin, and were blocked by PKA inhibitors. These results demonstrate that A2A receptor regulates a number of HSC fate decisions and induces greater HSC proliferation, reduces apoptosis and senescence by decreasing p53 and Rb through cAMP-PKA/Rac1/p38 MAPK pathway. This provides a mechanism for adenosine induced HSC regulation and liver fibrosis.


Psychiatric side effects during methysergide treatment.
A patient is reported with psychological change characterised by impaired concentration and thought projection, followed by both severe anxiety and depression, starting after three weeks on high dose methysergide. The acute problem settled slowly after methysergide withdrawal and is likely to represent an unusual and serious side effect of that drug.


An approach for modelling CO2 emissions from road traffic in urban areas.
An approach that incorporates three modelling components has been developed to estimate road traffic CO2 emissions for an urban area with street level resolution. The first component enables the determination of the road traffic characteristics using the SATURN (Simulation and Assignment of Traffic in Urban Road Networks) model. The output from this component is then analysed using MATrix LABoratory (MATLAB) programming to provide estimates of CO2 emissions for the urban area. Finally, ArcGIS is used to illustrate the model output. The three components are integrated using a Loose-Coupling approach in which the individual components each load the necessary data to give an independent output. The model structure is discussed in the current paper and the modelling results for a small city (Norwich, UK) are presented.


Mechanosensitivity of the cardiac muscarinic potassium channel. A novel property conferred by Kir3.4 subunit.
Muscarinic potassium channels are heterotetramers of Kir3.1 and other Kir3 channel subunits and play major roles in regulating membrane excitability in cardiac atrial, neuronal, and neuroendocrine tissues. We report here that rabbit atrial muscarinic potassium channels are rapidly and reversibly inhibited by membrane stretch, possibly serving as a mechanoelectrical feedback pathway. To probe the molecular basis for this phenomenon, we heterologously expressed heteromeric Kir3.1/Kir3.4 channels in Xenopus oocytes and found that they possess similar mechanosensitivity in response to hypo-osmolar stress. This could be attributed in part, if not exclusively, to the Kir3.4 subunit, which reproduced the mechanosensitivity of the heteromeric channel when expressed as a homomeric channel in oocytes. Kir3.4 is the first stretch-inactivated potassium channel to be identified molecularly. Physiologically, this feature may be important in atrial volume-sensing and other responses to stretch.


A case of multiple splenic abscesses managed non-operatively.
We report the case of a previously fit 22-year-old man who developed multiple splenic abscesses of unclear aetiology and was successfully treated with percutaneous drainage of the largest abscess and antibiotic therapy.


Training increases [3H]fucose incorporation in chick brain only if followed by memory storage.
Incorporation of [3H]fucose into glycoproteins is increased in the forebrain base and anterior forebrain roof of day-old chicks following passive avoidance training. In chicks trained but rendered apparently amnestic by immediate transcranial electroshock, the increased incorporation by comparison with untrained-shocked birds was abolished. If the electroshock was delayed by 10 min, the chicks showed recall for the task and fucose incorporation was increased by comparison with untrained but shocked or trained and immediate-shocked birds. We conclude that the increased incorporation of [3H]fucose is not a result of any concomitant of training but is directly associated with engram formation.


Single-channel kinetics of BK (Slo1) channels.
Single-channel kinetics has proven a powerful tool to reveal information about the gating mechanisms that control the opening and closing of ion channels. This introductory review focuses on the gating of large conductance Ca(2+)- and voltage-activated K(+) (BK or Slo1) channels at the single-channel level. It starts with single-channel current records and progresses to presentation and analysis of single-channel data and the development of gating mechanisms in terms of discrete state Markov (DSM) models. The DSM models are formulated in terms of the tetrameric modular structure of BK channels, consisting of a central transmembrane pore-gate domain (PGD) attached to four surrounding transmembrane voltage sensing domains (VSD) and a large intracellular cytosolic domain (CTD), also referred to as the gating ring. The modular structure and data analysis shows that the Ca(2+) and voltage dependent gating considered separately can each be approximated by 10-state two-tiered models with five closed states on the upper tier and five open states on the lower tier. The modular structure and joint Ca(2+) and voltage dependent gating are consistent with a 50 state two-tiered model with 25 closed states on the upper tier and 25 open states on the lower tier. Adding an additional tier of brief closed (flicker states) to the 10-state or 50-state models improved the description of the gating. For fixed experimental conditions a channel would gate in only a subset of the potential number of states. The detected number of states and the correlations between adjacent interval durations are consistent with the tiered models. The examined models can account for the single-channel kinetics and the bursting behavior of gating. Ca(2+) and voltage activate BK channels by predominantly increasing the effective opening rate of the channel with a smaller decrease in the effective closing rate. Ca(2+) and depolarization thus activate by mainly destabilizing the closed states.


A Rare Case of Gemcitabine-Induced Pulmonary Hypertension.
Gemcitabine is the backbone of systemic treatment of locally advanced and metastatic intrahepatic cholangiocarcinoma. In recent literature, gemcitabine has been linked to various pulmonary side effects. We report a case of an 82-year-old male who developed acute pulmonary hypertension after receiving one cycle of gemcitabine for metastatic cholangiocarcinoma. His symptoms began with fatigue associated with shortness of breath and cough that worsened despite dose reduction. He developed new onset bilateral pulmonary effusions and an echocardiogram revealed findings consistent with pulmonary hypertension. A computed tomography (CT) angiogram was negative for pulmonary thromboembolism. Although he was promptly treated with diuretics and steroids, the patient could not tolerate any further therapy. Gemcitabine-induced pulmonary hypertension is rare and can be challenging to diagnose, as it remains a diagnosis of exclusion. However, physicians should be vigilant of new pulmonary symptoms, as delayed treatment can cause significant patient morbidity and mortality.


Kinetic modeling of the assembly, dynamic steady state, and contraction of the FtsZ ring in prokaryotic cytokinesis.
Cytokinesis in prokaryotes involves the assembly of a polymeric ring composed of FtsZ protein monomeric units. The Z ring forms at the division plane and is attached to the membrane. After assembly, it maintains a stable yet dynamic steady state. Once induced, the ring contracts and the membrane constricts. In this work, we present a computational deterministic biochemical model exhibiting this behavior. The model is based on biochemical features of FtsZ known from in vitro studies, and it quantitatively reproduces relevant in vitro data. An essential part of the model is a consideration of interfacial reactions involving the cytosol volume, where monomeric FtsZ is dispersed, and the membrane surface in the cell's mid-zone where the ring is assembled. This approach allows the same chemical model to simulate either in vitro or in vivo conditions by adjusting only two geometrical parameters. The model includes minimal reactions, components, and assumptions, yet is able to reproduce sought-after in vivo behavior, including the rapid assembly of the ring via FtsZ-polymerization, the formation of a dynamic steady state in which GTP hydrolysis leads to the exchange of monomeric subunits between cytoplasm and the ring, and finally the induced contraction of the ring. The model gives a quantitative estimate for coupling between the rate of GTP hydrolysis and of FtsZ subunit turnover between the assembled ring and the cytoplasmic pool as observed. Membrane constriction is chemically driven by the strong tendency of GTP-bound FtsZ to self-assembly. The model suggests a possible mechanism of membrane contraction without a motor protein. The portion of the free energy of GTP hydrolysis released in cyclization is indirectly used in this energetically unfavorable process. The model provides a limit to the mechanistic complexity required to mimic ring behavior, and it highlights the importance of parallel in vitro and in vivo modeling.


The microstructure and bulk rheology of human cervicovaginal mucus are remarkably resistant to changes in pH.
The protective barrier, lubricant, and clearance functions of mucus are intimately coupled to its microstructure and bulk rheology. Mucus gels consist of a network of mucin biopolymers along with lipids, salts, and other proteins and exhibit similar biochemical and physical properties across diverse mucosal surfaces. Nevertheless, mucus is exposed to a broad range of pH values throughout the human body. Protein functions are typically sensitive to small changes in pH, and prior investigations using reconstituted, purified mucin gels suggested mucus undergoes a transition from a low-viscosity liquid at neutral pH to a highly viscoelastic solid at low pH. We sought to determine whether those observations hold for fresh, minimally perturbed human mucus ex vivo by using different-sized muco-inert nanoparticles to probe microstructure and cone-and-plate rheometry to measure bulk rheology. We demonstrate that both the microstructure and bulk rheology of fresh, undiluted, and minimally perturbed cervicovaginal mucus exhibit relatively minor changes from pH 1-2 to 8-9, in marked contrast with the pH sensitivity of purified mucin gels. Our work also suggests additional components in mucus secretions, typically eliminated during mucin purification and reconstitution, may play an important role in maintaining the protective properties of mucus.


Unpacking the mechanisms captured by a correlative species distribution model to improve predictions of climate refugia.
Climate refugia are regions that animals can retreat to, persist in and potentially then expand from under changing environmental conditions. Most forecasts of climate change refugia for species are based on correlative species distribution models (SDMs) using long-term climate averages, projected to future climate scenarios. Limitations of such methods include the need to extrapolate into novel environments and uncertainty regarding the extent to which proximate variables included in the model capture processes driving distribution limits (and thus can be assumed to provide reliable predictions under new conditions). These limitations are well documented; however, their impact on the quality of climate refugia predictions is difficult to quantify. Here, we develop a detailed bioenergetics model for the koala. It indicates that range limits are driven by heat-induced water stress, with the timing of rainfall and heat waves limiting the koala in the warmer parts of its range. We compare refugia predictions from the bioenergetics model with predictions from a suite of competing correlative SDMs under a range of future climate scenarios. SDMs were fitted using combinations of long-term climate and weather extremes variables, to test how well each set of predictions captures the knowledge embedded in the bioenergetics model. Correlative models produced broadly similar predictions to the bioenergetics model across much of the species' current range - with SDMs that included weather extremes showing highest congruence. However, predictions in some regions diverged significantly when projecting to future climates due to the breakdown in correlation between climate variables. We provide unique insight into the mechanisms driving koala distribution and illustrate the importance of subtle relationships between the timing of weather events, particularly rain relative to hot-spells, in driving species-climate relationships and distributions. By unpacking the mechanisms captured by correlative SDMs, we can increase our certainty in forecasts of climate change impacts on species.


In vivo imaging shows continued association of several IFT-A, IFT-B and dynein complexes while IFT trains U-turn at the tip.
Flagellar assembly depends on intraflagellar transport (IFT), a bidirectional motility of protein carriers, the IFT trains. The trains are periodic assemblies of IFT-A and IFT-B subcomplexes and the motors kinesin-2 and IFT dynein. At the tip, anterograde trains are remodeled for retrograde IFT, a process that in Chlamydomonas involves kinesin-2 release and train fragmentation. However, the degree of train disassembly at the tip remains unknown. Here, we performed two-color imaging of fluorescent protein-tagged IFT components, which indicates that IFT-A and IFT-B proteins from a given anterograde train usually return in the same set of retrograde trains. Similarly, concurrent turnaround was typical for IFT-B proteins and the IFT dynein subunit D1bLIC-GFP but severance was observed as well. Our data support a simple model of IFT turnaround, in which IFT-A, IFT-B and IFT dynein typically remain associated at the tip and segments of the anterograde trains convert directly into retrograde trains. Continuous association of IFT-A, IFT-B and IFT dynein during tip remodeling could balance protein entry and exit, preventing the build-up of IFT material in flagella.


Free radical-induced inactivation of creatine kinase: influence on the octameric and dimeric states of the mitochondrial enzyme (Mib-CK).
Free radicals of X-ray-induced water radiolysis, either directly or indirectly via their reaction products, reduce the activity of both dimeric cytoplasmic muscle-type creatine kinase (MM-CK) and octameric mitochondrial creatine kinase (Mi-CK) to virtually zero. Similarly values of the characteristic D(37)-dose of enzyme inactivation (dose required to reduce enzyme activity to 37%) were found for the two isoenzymes of CK under identical conditions. Octamer stability was not significantly affected within the dose range considered. However, both the dissociation of octamers into dimers by a transition-state analogue complex (TSAC), and the reassociation of the dimers into octamers, showed dose-dependent reduction. Binding of the TSAC to the active centre was found to protect the enzyme against inactivation by free radicals. No protection was observed for the radiation-induced decrease of the endogenous tryptophan fluorescence. The experimental results are in line with the following interpretation: (i) the reduction of Mi(b)-CK dimer association is due to free radical-induced modification of Trp-264, situated at the dimer/dimer interface; (ii) the active-site Trp-223 is not a prime target for free radicals and is not involved in the inactivation of the enzyme; (iii) the inhibition of TSAC-induced dissociation of Mi(b)-CK, like enzyme inactivation, is primarily due to a modification of the active-site Cys-278.


The oocyte: the key player in the success of assisted reproduction technologies.
The ovulation of a mature oocyte at metaphase II of meiosis, with optimal potential to undergo fertilisation by a sperm cell, complete meiosis and sustain the switch to mitotic division, and support early embryo development, involves a protracted and disrupted/delayed series of processes. Many of these are targeted for exploitation in vivo , or recapitulation in vitro , by the livestock industry. Reproductive technologies, including AI, multiple ovulation embryo transfer, ovum pick-up, in vitro embryo production, and oestrus and ovulation synchronisation, offer practitioners and producers the opportunity to produce offspring from genetically valuable dams in much greater numbers than they would normally have in their lifetime, while in vitro oocyte and follicle culture are important platforms for researchers to interrogate the physiological mechanisms driving fertility. The majority of these technologies target the ovarian follicle and the oocyte within; thus, the quality and capability of the recovered oocyte determine the success of the reproductive intervention. Molecular and microscopical technologies have grown exponentially, providing powerful platforms to interrogate the molecular mechanisms which are integral to or affected by ART. The development of the bovine oocyte from its differentiation in the ovary to ovulation is described in the light of its relevance to key aspects of individual interventions, while highlighting the historical timeline.


Silage or limit-fed grain growing diets for steers: I. Growth and carcass quality.
The influence of energy source (silage- [S] or grain- [G] based) on organ growth, carcass quality, and meat acceptability independent of rate of gain was examined. Sixty-four Angus steers were allotted to one of the two treatments and given ad libitum access to silage or limit-fed grain for 145 d. All steers were then given ad libitum access to a grain diet for 45, 75, or 105 d. Eight steers from each treatment were slaughtered at the end of the growing phase and at each of the termination dates. The silage-based growing diet consisted (DM basis) of 55% sorghum silage (averaged 23.6% dry matter), 22% alfalfa hay, 10.8% ground shelled corn, and 10.8% soybean meal and contained 12.8% CP. Dry matter in the grain-based diet, composed of 76.5% ground shelled corn, 5% soybean meal, 13.6% cottonseed hulls, 3.5% molasses, and .4% salt and 1% limestone, contained 12.1% CP. It was limit-fed to produce rates of gain similar to the silage diet eaten ad libitum, using net energy for gain of each diet calculated from organic matter digestibility determined in digestion trials. The finishing diet was similar to the grain growing diet except that alfalfa hay replaced the cottonseed hulls. No implants or ionophores were used. High silage moisture decreased ADG the first 45 d, so steers fed grain gained faster, but thereafter gains were similar. At the end of the growing phase, steers fed grain had heavier shrunk and empty body weights and larger livers. However, liver size was not different when adjusted for growing ADG. By 45 d with ad libitum access to the finishing diet, 75% of the carcasses from steers fed both diets graded Choice. Steers fed silage had tougher (P < .05) steaks with less flavor intensity (P < .05) at the end of the growing phase; these differences diminished after 75 d on feed. These results suggest that choice beef can be produced in only 45 d in the feedlot, but tenderness and flavor among Choice carcasses remained inferior for steers fed silage for at least 75 d on a high-grain diet.


Real-World Adherence in Patients with Metastatic Colorectal Cancer Treated with Trifluridine plus Tipiracil or Regorafenib.
Trifluridine and tipiracil (FTD + TPI) and regorafenib (REG) are approved treatments for the treatment of refractory metastatic colorectal cancer (mCRC). This study assesses adherence and duration of therapy with FTD + TPI versus REG and explores the effect of sequencing on adherence. Adults diagnosed with mCRC were identified in the IQVIA Real-World Data Adjudicated Claims: U.S. database (October 2014-July 2017). The observation period spanned from the index date (first dispensing of FTD + TPI or REG) to the earliest of a switch to another mCRC agent, the end of continuous enrollment, or the end of data availability. Medication possession ratio (MPR), proportion of days covered (PDC), and persistence and time to discontinuation (gap ≥45 days) were compared between FTD + TPI and REG users and among switchers (FTD + TPI-to-REG vs. REG-to-FTD + TPI). A total of 469 FTD + TPI and 311 REG users were identified. FTD + TPI users had higher compliance with an MPR ≥80% (odds ratio [OR], 2.47; p < .001) and PDC ≥80% (OR, 2.77; p < .001). FTD + TPI users had better persistence (82.8% vs. 68.0%; p < .001) and lower risk of discontinuation (hazard ratio [HR], 0.76; p = .006). Among switchers (96 FTD + TPI-to-REG; 83 REG-to-FTD + TPI), those switching from FTD + TPI to REG were more likely to have an MPR ≥80% (OR, 2.91; p < .001) and PDC ≥80% (OR, 4.60; p < .001) compared with REG-to-FTD + TPI switchers while treated with these drugs. Additionally, FTD + TPI-to-REG switchers had a lower risk of first treatment discontinuation (HR, 0.66; p = .009). FTD + TPI users had significantly higher adherence and persistence, and patients who were treated with FTD + TPI before switching to REG also had higher adherence and persistence outcomes. Trifluridine plus tipiracil (FTD + TPI) and regorafenib (REG) prolong survival in refractory metastatic colorectal cancer (mCRC) but have different tolerability profiles. This study assessed real-world adherence to treatment with FTD + TPI versus REG and compared outcomes among patients who switched from FTD + TPI to REG and vice versa. FTD + TPI was associated with significantly higher medication adherence and longer time to discontinuation than REG. Patients treated with FTD + TPI prior to switching to REG also showed higher adherence outcomes. Findings could help inform decision making regarding the choice and sequencing of treatment with FTD + TPI versus REG in patients with mCRC.


(Hyper)active Data Curation: A Video Case Study from Behavioral Science.
Video data are uniquely suited for research reuse and for documenting research methods and findings. However, curation of video data is a serious hurdle for researchers in the social and behavioral sciences, where behavioral video data are obtained session by session and data sharing is not the norm. To eliminate the onerous burden of post hoc curation at the time of publication (or later), we describe best practices in active data curation-where data are curated and uploaded immediately after each data collection to allow instantaneous sharing with one button press at any time. Indeed, we recommend that researchers adopt "hyperactive" data curation where they openly share every step of their research process. The necessary infrastructure and tools are provided by Databrary-a secure, web-based data library designed for active curation and sharing of personally identifiable video data and associated metadata. We provide a case study of hyperactive curation of video data from the Play and Learning Across a Year (PLAY) project, where dozens of researchers developed a common protocol to collect, annotate, and actively curate video data of infants and mothers during natural activity in their homes at research sites across North America. PLAY relies on scalable standardized workflows to facilitate collaborative research, assure data quality, and prepare the corpus for sharing and reuse throughout the entire research process.


Direct detection of bacterial biofilms on the middle-ear mucosa of children with chronic otitis media.
Chronic otitis media (OM) is a common pediatric infectious disease. Previous studies demonstrating that metabolically active bacteria exist in culture-negative pediatric middle-ear effusions and that experimental infection with Haemophilus influenzae in the chinchilla model of otitis media results in the formation of adherent mucosal biofilms suggest that chronic OM may result from a mucosal biofilm infection. To test the hypothesis that chronic OM in humans is biofilm-related. Middle-ear mucosa (MEM) biopsy specimens were obtained from 26 children (mean age, 2.5 [range, 0.5-14] years) undergoing tympanostomy tube placement for treatment of otitis media with effusion (OME) and recurrent OM and were analyzed using microbiological culture, polymerase chain reaction (PCR)-based diagnostics, direct microscopic examination, fluorescence in situ hybridization, and immunostaining. Uninfected (control) MEM specimens were obtained from 3 children and 5 adults undergoing cochlear implantation. Patients were enrolled between February 2004 and April 2005 from a single US tertiary referral otolaryngology practice. Confocal laser scanning microscopic (CLSM) images were obtained from MEM biopsy specimens and were evaluated for biofilm morphology using generic stains and species-specific probes for H influenzae, Streptococcus pneumoniae, and Moraxella catarrhalis. Effusions, when present, were evaluated by PCR and culture for evidence of pathogen-specific nucleic acid sequences and bacterial growth, respectively. Of the 26 children undergoing tympanostomy tube placement, 13 (50%) had OME, 20 (77%) had recurrent OM, and 7 (27%) had both diagnoses; 27 of 52 (52%) of the ears had effusions, 24 of 24 effusions were PCR-positive for at least 1 OM pathogen, and 6 (22%) of 27 effusions were culture-positive for any pathogen. Mucosal biofilms were visualized by CLSM on 46 (92%) of 50 MEM specimens from children with OME and recurrent OM using generic and pathogen-specific probes. Biofilms were not observed on 8 control MEM specimens obtained from the patients undergoing cochlear implantation. Direct detection of biofilms on MEM biopsy specimens from children with OME and recurrent OM supports the hypothesis that these chronic middle-ear disorders are biofilm-related.


Zirconium ion mediated formation of liposome multilayers.
Phospholipid vesicles have attracted considerable interest as a platform for a variety of biomolecular binding assays, especially in the area of membrane protein sensing. The development of liposome-based biosensors widely relies on the availability of simple and efficient protocols for their surface immobilization. We present a novel approach toward the creation of three-dimensional phospholipid vesicle constructs using multivalent zirconium ions as linkers between the liposomes. Such three-dimensional sensing platforms are likely to play a key role in the development of biosensing devices with increased loading capacity and sensitivity. After demonstrating the affinity of Zr(4+) toward the phospholipids, we formed vesicle multilayers by sequential injections of solutions containing either liposomes or ZrOCl(2). In situ adlayer characterization was carried out by optical waveguide lightmode spectroscopy (OWLS) and quartz crystal microbalance with dissipation (QCM-D) measurements while imaging was performed by atomic force microscopy (AFM) and fluorescence microscopy. Multilayers were successfully constructed, and as demonstrated in a model fluorescence-based biomolecular binding assay, the sensor's loading capacity was increased. Furthermore, we observed that lipid exchange between the vesicles is promoted in the presence of Zr(4+) and that addition of a phosphate-containing buffer leads to adlayer loosening and creation of lipidic tubular structures. The approach presented here could be applied to the study of membrane proteins in a highly sensitive manner due to the increased surface area or to produce functional coatings for controlled drug release and host response.


N-terminal pro-B-type natriuretic peptide concentrations predict the risk of cardiovascular adverse events from antiinflammatory drugs: a pilot trial.
we investigated whether higher concentrations of N-terminal pro-B-type natriuretic peptide (NT-proBNP) predicts cardiovascular adverse events (CV-AEs) in patients with osteoarthritis treated with antiinflammatory drugs. NT-proBNP was measured in baseline samples from 433 patients enrolled in a prospective randomized study designed to test the therapeutic effect of a novel metalloproteinase inhibitor. We monitored CV-AEs and retrospectively investigated their relationship to the concomitant use of selective cyclooxygenase-2 inhibitors (coxibs), traditional nonsteroidal antiinflammatory drugs (tNSAIDs), and glucocorticoids. CV-AEs included myocardial infarction, stroke, new or worsening of preexisting arterial hypertension, congestive heart failure, and several less severe CV-AEs. we observed 82 mild to serious CV-AEs during an observational period of 200 days. The risk of such events was 1.95-fold higher in patients who were taking tNSAIDs, glucocorticoids, or coxibs (i.e., any inhibitor) and who had NT-proBNP concentrations > or = 100 ng/L than in patients taking any inhibitor who had NT-proBNP values <100 ng/L (P < 0.05). Patients taking coxibs (alone or in addition to tNSAIDs or glucocorticoids) with baseline NT-proBNP values > or = 100 ng/L had a 7.41-fold higher risk for CV-AEs than those with baseline values <100 ng/L (P < 0.01). Patients who were taking 2 or more antiinflammatory drugs and had NT-proBNP values > or = 100 ng/L had a 3.74-fold higher risk for CV-AEs than those with NT-proBNP values <100 ng/L (P < 0.05). An NT-proBNP value <100 ng/L was associated with negative predictive values of >85% across all treatment groups. NT-proBNP may be a useful marker for anticipating cardiovascular risk associated with the use of antiinflammatory drugs for osteoarthritis.


Controlled clinical comparison of BACTEC plus anaerobic/F to standard anaerobic/F as the anaerobic companion bottle to plus aerobic/F medium for culturing blood from adults.
To determine the optimal anaerobic companion bottle to pair with BACTEC Plus Aerobic/F medium for recovery of pathogenic microorganisms from adult patients with bacteremia and fungemia, we compared Plus Anaerobic/F bottles with Standard Anaerobic/F bottles, each of which was filled with 4 to 6 ml of blood. The two bottles were paired with a Plus Aerobic/F bottle filled with 8 to 12 ml of blood. A total of 14,011 blood culture sets were obtained. Of these, 11,583 sets were received with all three bottles filled adequately and 12,257 were received with both anaerobic bottles filled adequately. Of 818 clinically important isolates detected in one or both adequately filled anaerobic bottles, significantly more staphylococci (P < 0.001), streptococci (P < 0.005), Escherichia coli isolates (P < 0.02), Klebsiella pneumoniae isolates (P < 0.005), and all microorganisms combined (P < 0.001) were detected in Plus Anaerobic/F bottles. In contrast, significantly more anaerobic gram-negative bacilli were detected in Standard Anaerobic/F bottles (P < 0.05). Of 397 unimicrobial episodes of septicemia, 354 were detected with both pairs, 30 were detected with Plus Aerobic/F-Plus Anaerobic/F pairs only, and 13 were detected with Plus Aerobic/F-Standard Anaerobic/F pairs only (P < 0.05). Significantly more episodes of bacteremia caused by members of the family Enterobacteriaceae (P < 0.05) and aerobic and facultative gram-positive bacteria (P < 0.025) were detected with Plus Anaerobic/F bottles only. In a paired-bottle analysis, 810 of 950 isolates were recovered from both pairs, 90 were recovered from Plus Aerobic/F-Plus Anaerobic/F pairs only, and 50 were recovered from Plus Aerobic/F-Standard Anaerobic/F pairs only (P < 0.001). Paired Plus Aerobic/F-Plus Anaerobic/F bottles yielded significantly more staphylococci (P < 0.001), streptococci (P < 0.05), and members of the family Enterobacteriaceae (P <0.001). We conclude that Plus Anaerobic/F bottles detect more microorganisms and episodes of bacteremia and fungemia than Standard Anaerobic/F bottles as companion bottles to Plus Aerobic/F bottles in the BACTEC 9240 blood culture system.


Role of ferritin in the control of the labile iron pool in murine erythroleukemia cells.
In vitro studies have shown that ferritin iron incorporation is mediated by a ferroxidase activity associated with ferritin H subunits (H-Ft) and a nucleation center associated with ferritin L subunits (L-Ft). To assess the role played by the ferritin subunits in regulating intracellular iron distribution, we transfected mouse erythroleukemia cells with the H-Ft subunit gene mutated in the iron-responsive element. Stable transfectants displayed high H-Ft levels and reduced endogenous L-Ft levels, resulting in a marked change in the H:L subunit ratio from 1:1 in control cells to as high as 20:1 in some transfected clones. The effects of H-Ft overexpression on the labile iron pool were determined in intact cells by a novel method based on the fluorescent metallosensor calcein. H-Ft overexpression resulted in a significant reduction in the iron pool, from 1.3 microM in control cells to 0.56 microM in H-Ft transfectants, and in higher buffering capacity following iron loads. A fraction of the H-Ft-associated iron was labile, available to cell-permeant, but not cell-impermeant, chelators. The results of this study provide the first in vivo direct demonstration of the capacity of H-Ft to sequester cell iron and to regulate the levels of the labile iron pool.


Acute mental health service use following onset of the COVID-19 pandemic in Ontario, Canada: a trend analysis.
The extent to which heightened distress during the COVID-19 pandemic translated to increases in severe mental health outcomes is unknown. We examined trends in psychiatric presentations to acute care settings in the first 12 months after onset of the pandemic. This was a trends analysis of administrative population data in Ontario, Canada. We examined rates of hospitalizations and emergency department visits for mental health diagnoses overall and stratified by sex, age and diagnostic grouping (e.g., mood disorders, anxiety disorders, psychotic disorders), as well as visits for intentional self-injury for people aged 10 to 105 years, from January 2019 to March 2021. We used Joinpoint regression to identify significant inflection points after the onset of the pandemic in March 2020. Among the 12 968 100 people included in our analysis, rates of mental health-related hospitalizations and emergency department visits declined immediately after the onset of the pandemic (peak overall decline of 30% [hospitalizations] and 37% [emergency department visits] compared to April 2019) and returned to near prepandemic levels by March 2021. Compared to April 2019, visits for intentional self-injury declined by 33% and remained below prepandemic levels until March 2021. We observed the largest declines in service use among adolescents aged 14 to 17 years (55% decline in hospitalizations, 58% decline in emergency department visits) and 10 to 13 years (56% decline in self-injury), and for those with substance-related disorders (33% decline in emergency department visits) and anxiety disorders (61% decline in hospitalizations). Contrary to expectations, the abrupt decline in acute mental health service use immediately after the onset of the pandemic and the return to near prepandemic levels that we observed suggest that changes and stressors in the first 12 months of the pandemic did not translate to increased service use. Continued surveillance of acute mental health service use is warranted.


Range dynamics, rather than convergent selection, explain the mosaic distribution of red-winged blackbird phenotypes.
Geographic distributions of genetic and phenotypic characters can illuminate historical evolutionary processes. In particular, mosaic distributions of phenotypically similar populations can arise from parallel evolution or from irregular patterns of dispersal and colonization by divergent forms. Two phenotypically divergent forms of the red-winged blackbird (Agelaius phoeniceus) show a mosaic phenotypic distribution, with a "bicolored" form occurring disjunctly in California and Mexico. We analyzed the relationships among these bicolored populations and neighboring typical populations, using ∼600 bp of mitochondrial DNA sequence data and 10 nuclear short tandem repeat loci. We find that bicolored populations, although separated by ∼3000 km, are genetically more similar to one other than they are to typical populations separated by ∼400 km. We also find evidence of ongoing gene flow among populations, including some evidence of asymmetric gene flow. We conclude that the current distribution of bicolored forms represents incomplete speciation, where recent asymmetric hybridization with typical A. phoeniceus is dividing the range of a formerly widespread bicolored form. This hypothesis predicts that bicolored forms may suffer extinction by hybridization. Future work will use fine-scaled geographical sampling and nuclear sequence data to test for hybrid origins of currently typical populations and to more precisely quantify the directionality of gene flow.


Differential responses of abaxial and adaxial guard cells of broad bean to abscisic acid and calcium
Regulation by abscisic acid (ABA) and Ca2+ of broad bean (Vicia faba) abaxial and adaxial guard cell movements and inward K+ currents were compared. One millimolar Ca2+ in the bathing medium inhibited abaxial stomatal opening by 60% but only inhibited adaxial stomatal opening by 15%. The addition of 1 &mgr;M ABA in the bathing medium resulted in 80% inhibition of abaxial but only 45% inhibition of adaxial stomatal opening. Similarly, ABA and Ca2+ each stimulated greater abaxial stomatal closure than adaxial stomatal closure. Whole-cell patch-clamp results showed that the inward K+ currents of abaxial guard cells were inhibited by 60% (-180 mV) in the presence of 1.5 &mgr;M Ca2+ in the cytoplasm, whereas the inward K+ currents of adaxial guard cells were not affected at all by the same treatment. Although 1 &mgr;M ABA in the cytoplasm inhibited the inward K+ currents to a similar extent for both abaxial and adaxial guard cells, the former were more sensitive to ABA applied externally. These results suggest that the abaxial stomata are more sensitive to Ca2+ and ABA than adaxial stomata in regard to stomatal opening and closing processes and that the regulation of the inward K+ currents by ABA may not proceed via a Ca2+-signaling pathway in adaxial guard cells. Therefore, there may be different pathways for ABA- and Ca2+-mediated signal transduction in abaxial and adaxial guard cells.


On-demand tuned hazard free elastomeric composites: A green approach.
Rising ecological concerns and depletion of the potentially harmful environmental impacts caused by rubber products, are of prime importance in the industry. Therefore, implementation of sustainable greener materials is required to minimize the detrimental influences. In this research, we investigated the beneficial influence of naturally derived bio-resin toward the effects of association with Zinc Oxide Nanoparticles in highly dispersible silica (HDS) reinforced Natural rubber (NR)/Epoxidized Natural Rubber (ENR)-based composites. This novel green composite offers impressive properties which were analyzed based on bound rubber content, transmission electron microscopy, physico-mechanical, dynamic mechanical, and cure characteristics. Nanoindentation studies demonstrated the enhanced hysteresis phenomenon of the green composites. The small angle X-ray scattering (SAXS) characterization has been studied by using a Beaucage model and results corroborates that the insertion of bio-resin exhibits ameliorated state of silica dispersion in the green composites. Overall, the study with the bio-resin has provided the impetus in employing it as an alternative to the expensive synthetic route of silane coupling agent and toxic process oil.


Measuring childhood maltreatment to predict early-adult psychopathology: Comparison of prospective informant-reports and retrospective self-reports.
Both prospective informant-reports and retrospective self-reports may be used to measure childhood maltreatment, though both methods entail potential limitations such as underestimation and memory biases. The validity and utility of standard measures of childhood maltreatment requires clarification in order to inform the design of future studies investigating the mental health consequences of maltreatment. The present study assessed agreement between prospective informant-reports and retrospective self-reports of childhood maltreatment, as well as the comparative utility of both reports for predicting a range of psychiatric problems at age 18. Data were obtained from the Environmental Risk (E-Risk) Longitudinal Twin Study, a nationally-representative birth cohort of 2232 children followed to 18 years of age (with 93% retention). Childhood maltreatment was assessed in two ways: (i) prospective informant-reports from caregivers, researchers, and clinicians when children were aged 5, 7, 10 and 12; and (ii) retrospective self-reports of maltreatment experiences occurring up to age 12, obtained at age 18 using the Childhood Trauma Questionnaire. Participants were privately interviewed at age 18 concerning several psychiatric problems including depression, anxiety, self-injury, alcohol/cannabis dependence, and conduct disorder. There was only slight to fair agreement between prospective and retrospective reports of childhood maltreatment (all Kappa's ≤ 0.31). Both prospective and retrospective reports of maltreatment were associated with age-18 psychiatric problems, though the strongest associations were found when maltreatment was retrospectively self-reported. These findings indicate that prospective and retrospective reports of childhood maltreatment capture largely non-overlapping groups of individuals. Young adults who recall being maltreated have a particularly elevated risk for psychopathology.


Validity of measures of pain and symptoms in HIV/AIDS infected households in resources poor settings: results from the Dominican Republic and Cambodia.
HIV/AIDS treatment programs are currently being mounted in many developing nations that include palliative care services. While measures of palliative care have been developed and validated for resource rich settings, very little work exists to support an understanding of measurement for Africa, Latin America or Asia. This study investigates the construct validity of measures of reported pain, pain control, symptoms and symptom control in areas with high HIV-infected prevalence in Dominican Republic and Cambodia Measures were adapted from the POS (Palliative Outcome Scale). Households were selected through purposive sampling from networks of people living with HIV/AIDS. Consistencies in patterns in the data were tested used Chi Square and Mantel Haenszel tests. The sample persons who reported chronic illness were much more likely to report pain and symptoms compared to those not chronically ill. When controlling for the degrees of pain, pain control did not differ between the chronically ill and non-chronically ill using a Mantel Haenszel test in both countries. Similar results were found for reported symptoms and symptom control for the Dominican Republic. These findings broadly support the construct validity of an adapted version of the POS in these two less developed countries. The results of the study suggest that the selected measures can usefully be incorporated into population-based surveys and evaluation tools needed to monitor palliative care and used in settings with high HIV/AIDS prevalence.


A novel mode of chromosomal evolution peculiar to filamentous Ascomycete fungi.
Gene loss, inversions, translocations, and other chromosomal rearrangements vary among species, resulting in different rates of structural genome evolution. Major chromosomal rearrangements are rare in most eukaryotes, giving large regions with the same genes in the same order and orientation across species. These regions of macrosynteny have been very useful for locating homologous genes in different species and to guide the assembly of genome sequences. Previous analyses in the fungi have indicated that macrosynteny is rare; instead, comparisons across species show no synteny or only microsyntenic regions encompassing usually five or fewer genes. To test the hypothesis that chromosomal evolution is different in the fungi compared to other eukaryotes, synteny was compared between species of the major fungal taxa. These analyses identified a novel form of evolution in which genes are conserved within homologous chromosomes, but with randomized orders and orientations. This mode of evolution is designated mesosynteny, to differentiate it from micro- and macrosynteny seen in other organisms. Mesosynteny is an alternative evolutionary pathway very different from macrosyntenic conservation. Surprisingly, mesosynteny was not found in all fungal groups. Instead, mesosynteny appears to be restricted to filamentous Ascomycetes and was most striking between species in the Dothideomycetes. The existence of mesosynteny between relatively distantly related Ascomycetes could be explained by a high frequency of chromosomal inversions, but translocations must be extremely rare. The mechanism for this phenomenon is not known, but presumably involves generation of frequent inversions during meiosis.


Mucin production by Staphylococcus epidermidis. A virulence factor promoting adherence to vascular grafts.
The production of an exopolysaccharide (mucin) by some Staphylococcus epidermidis strains facilitates bacterial adhesion to prosthetic vascular grafts and may play an important role in adherence-mediated growth. An in vitro model was developed to measure the differential adherence of mucin-producing (RP-12) and nonmucin-producing (SP-2) S epidermidis strains onto expanded polytef and velour knitted Dacron graft material. After incubation in a 10(7)/mL suspension of organisms, graft specimens were repeatedly washed to remove nonadherent organisms and then sonicated to dislodge adherent organisms. Bacterial adherence was calculated from a quantitative culture of the sonication effluent. Both S epidermidis strains adhered in ten to 100 times greater numbers to the knitted Dacron graft material than to expanded polytef. The production of an exopolysaccharide by the RP-12 strain significantly increased adherence to both graft types compared with the SP-2 strain. The increased adherence of the RP-12 strain was inhibited by adding D-mannosamine to the inoculum. The in vitro model developed is well suited for further study of the mechanisms by which bacteria adhere to and colonize vascular grafts.


Mixed encounters, limited perception and optimal foraging.
This article demonstrates how perceptual constraints of predators and the possibility that predators encounter prey both sequentially (one prey type at a time) and simultaneously (two or more prey types at a time) may influence the predator attack decisions, diet composition and functional response of a behavioural predator-prey system. Individuals of a predator species are assumed to forage optimally on two prey types and to have exact knowledge of prey population numbers (or densities) only in a neighbourhood of their actual spatial location. The system characteristics are inspected by means of a discrete-time, discrete-space, individual-based model of the one-predator-two-prey interaction. Model predictions are compared with ones that have been obtained by assuming only sequential encounters of predators with prey and/or omniscient predators aware of prey population densities in the whole environment. It is shown that the zero-one prey choice rule, optimal for sequential encounters and omniscient predators, shifts to abruptly changing partial preferences for both prey types in the case of omniscient predators faced with both types of prey encounters. The latter, in turn, become gradually changing partial preferences when predator omniscience is considered only local.


Glyphosate applied at low doses can stimulate plant growth.
Glyphosate blocks the shikimic acid pathway, inhibiting the production of aromatic amino acids and several secondary compounds derived from these amino acids. Non-target plants can be exposed to low doses of glyphosate by herbicide drift of spray droplets and contact with treated weeds. Previous studies have reported that low doses of glyphosate stimulate growth, although these data are very limited. The objective of this study was to determine the effects of low glyphosate doses on growth of a range of plant species. Growth of maize, conventional soybean, Eucalyptus grandis Hill ex Maiden, Pinus caribea L. and Commelia benghalensis L. was enhanced by 1.8-36 g glyphosate ha(-1). Growth of glyphosate-resistant soybean was unaffected by any glyphosate dose from 1.8 to 720 g AE ha(-1). The optimum doses for growth stimulation were distinct for plant species and tissue evaluated. The greatest stimulation of growth was observed for C. benghalensis and P. caribea. Shikimic acid levels in tissues of glyphosate-treated soybean and maize were measured and found to be elevated at growth-stimulating doses. Subtoxic doses of glyphosate stimulate the growth of a range of plant species, as measured in several plant organs. This hormesis effect is likely to be related to the molecular target of glyphosate, since the effect was not seen in glyphosate-resistant plants, and shikimate levels were enhanced in plants with stimulated growth.


In vitro susceptibility characteristics of Cryptococcus neoformans varieties from AIDS patients in Goiânia, Brazil.
Sixty clinical isolates of Cryptococcus neoformans from AIDS from Goiânia, state of Goiás, Brazil, were characterized according to varieties, serotypes and tested for antifungal susceptibility. To differentiate the two varieties was used L-canavanine-glycine-bromothymol blue medium and to separate the serotypes was used slide agglutination test with Crypto Check Iatron. The Minimal Inhibitory Concentration (MIC) of fluconazole, itraconazole, and amphotericin B were determined by the National Committee for Clinical Laboratory Standards macrodilution method. Our results identified 56 isolates as C. neoformans var. neoformans serotype A and 4 isolates as C. neoformans var. gattii serotype B. MIC values for C. neoformans var. gattii were higher than C. neoformans var. neoformans. We verified that none isolate was resistant to itraconazole and to amphotericin B, but one C. neoformans var. neoformans and three C. neoformans var. gattii isolates were resistant to fluconazole. The presence of C. neoformans var. gattii fluconazole resistant indicates the importance of determining not only the variety of C. neoformans infecting the patients but also measuring the MIC of the isolate in order to properly orient treatment.


Synthesis of tetraphosphine macrocycles using copper(i) templates.
The synthesis of phosphine macrocycles is a relatively underdeveloped area and no standard synthetic routes have emerged. Accordingly, two general synthetic routes to tetradentate phosphine macrocycles were investigated. Both routes use Cu(i) ions as template ions because, unlike other metals such as Pd(ii) and Pt(ii), the Cu(i) ions can be removed from the macrocyclic complex without degrading the macrocycle ligand. The first route involves the coupling of two bidentate secondary phosphines bonded to Cu(i) using 1,3-dibromopropane or 1,4-dibromobutane. Using this route, tetradentate phosphine macrocycles with either -(CH2)3OCH3 or Ph groups bonded to the P atoms were synthesized. Macrocycle phosphines containing the -(CH2)3OCH3 groups were investigated for their potential water-solubility, but experiments showed these phosphines were not water soluble. The second synthetic route involved the alkylation of an open-chain, mixed tertiary-secondary, tetradentate phosphine coordinated to Cu(i). Following formation of the macrocyclic ligand, the Cu(i) template was removed by reaction with aqueous KCN to yield the free macrocyclic phosphine. This route was demonstrated for the preparation of the macrocyclic phosphine ligand 1,5,9,13-tetraphenyl-1,5,9,13-tetraphosphacycloheptadecane. Following demetallation, this macrocyclic ligand was coordinated to Fe(ii) and Co(ii) to form the corresponding macrocyclic phosphine complexes.


Comparative in vitro susceptibility of Bacteroides and Fusobacterium isolated from footrot in sheep to 28 antimicrobial agents.
The agar dilution method was used to determine the bacteriostatic activity of 28 antimicrobial agents against 141 strains to the genus Bacteroides and 29 strains from the genus Fusobacterium. All organisms were isolated from clinical cases of ovine footrot. The strains were isolated from 125 Merino sheep, over a period of 2 years, from January 1987 to December 1988. The three ureidopenicillins studied (azlocillin, mezlocillin and piperacillin) proved to be the most effective antimicrobial agents. Chloramphenicol, metronidazole and tinidazole effectively inhibited the growth of Bacteroides spp., while phosphomycin was active against Fusobacterium spp.


The fusiform face area subserves face perception, not generic within-category identification.
The function of the fusiform face area (FFA), a face-selective region in human extrastriate cortex, is a matter of active debate. Here we measured the correlation between FFA activity measured by functional magnetic resonance imaging (fMRI) and behavioral outcomes in perceptual tasks to determine the role of the FFA in the detection and within-category identification of faces and objects. Our data show that FFA activation is correlated on a trial-by-trial basis with both detecting the presence of faces and identifying specific faces. However, for most non-face objects (including cars seen by car experts), within-category identification performance was correlated with activation in other regions of the ventral occipitotemporal cortex, not the FFA. These results indicate that the FFA is involved in both detection and identification of faces, but that it has little involvement in within-category identification of non-face objects (including objects of expertise).


Evidence for competition and cooperation among climbing plants.
A plant's best strategy for acquiring resources may often depend on the identity of neighbours. Here, I ask whether plants adjust their strategy to local relatedness: individuals may cooperate (reduce competitiveness) with kin but compete relatively intensely with non-kin. In a greenhouse experiment with Ipomoea hederacea, neighbouring siblings from the same inbred line were relatively uniform in height; groups of mixed lines, however, were increasingly variable as their mean height increased. The reproductive yield of mixed and sibling groups was similar overall, but when adjusted to a common mean height and height inequality, the yield of mixed groups was significantly less. Where this difference in yield was most pronounced (among groups that varied most in height), mixed groups tended to allocate more mass to roots than comparable sibling groups, and overall, mixed groups produced significantly fewer seeds per unit mass of roots. These results suggest that, from the group perspective, non-kin may have wasted resources in below-ground competition at the expense of reproduction; kin groups, on the other hand, displayed the relative efficiency that is expected of reduced competitiveness.


Impact of Provider Imaging Practices on Survival Outcomes in Advanced Ovarian Cancer.
This study sought to describe how high- versus low-frequency surveillance imaging practices among providers at Memorial Sloan Kettering Cancer Center (MSKCC) impact overall survival (OS) and time to recurrence of patients with advanced epithelial ovarian cancer in first remission. The study cohort included patients with stage II-IV high-grade epithelial ovarian cancer diagnosed in January 2001 through January 2017 who experienced recurrence after initial platinum-based chemotherapy. To determine usual imaging practices for providers at MSKCC, median frequency of CT or MRI of the abdomen/pelvis was calculated among patients with a long-term remission (defined as at least 1 year) treated by each provider. Cox proportional hazards models were used to examine differences in OS and time to recurrence among patients treated by providers with high versus low imaging frequency practices, with additional subgroup analysis among patients with elevated CA-125 levels >35 U/mL at diagnosis. Chi-square tests were used to examine differences in the proportion of patients who enrolled in clinical trials or underwent secondary cytoreductive surgery (SCS) by imaging frequency. A total of 543 patients were treated by providers with high imaging frequency (>1 scan every 12 months) and 141 were treated by providers with low imaging frequency (≤1 scan every 12 months). Time to recurrence was shorter among patients treated by providers with high versus low imaging frequency (18.0 vs 19.2 months; hazard ratio, 1.33; P=.003). Results were similar when restricted to patients with elevated CA-125 levels at diagnosis. There was no significant difference in OS, clinical trial enrollment, or SCS by imaging practice. Within the limitations of this retrospective analysis, patients with advanced ovarian cancer treated by high-frequency-imaging providers had earlier detection of recurrence. Future analyses in a larger population are warranted to elucidate the risks versus benefits of surveillance imaging.


Climate model studies of synchronously rotating planets.
M stars constitute 75% of main sequence stars though, until recently, their star systems have not been considered suitable places for habitable planets to exist. In this study the climate of a synchronously rotating planet around an M dwarf star is evaluated using a three-dimensional global atmospheric circulation model. The presence of clouds and evaporative cooling at the surface of the planet result in a cooler surface temperature at the subsolar point. Water ice forms at the polar regions and on the dark side, where the minimum temperature lies between -30 degrees C and 0 degrees C. As expected, rainfall is extremely high on the starlit side and extremely low on the dark side. The presence of a dry continent causes higher temperatures on the dayside, and allows accumulation of snow on the nightside. The absence of any oceans leads to higher day-night temperature differences, consistent with previous work. The present study reinforces recent conclusions that synchronously rotating planets within the circumstellar habitable zones of M dwarf stars should be habitable, and therefore M dwarf systems should not be excluded in future searches for exoplanets.


Fibrosing inflammatory pseudotumors involving the skull base: MR and CT manifestations with histopathologic comparison.
To describe the MR and CT features of fibrosing inflammatory pseudotumors of the skull base region, and to document the MR signal intensity of the lesions with histopathologic comparison. We reviewed the MR and CT studies of five patients with pathologically proved fibrosing inflammatory pseudotumor involving the skull base. Unenhanced spin-echo T1- and T2-weighted and contrast-enhanced T1-weighted MR images were obtained at 0.5 T in three patients and at 1.5 T in two patients. MR findings were correlated with histopathologic findings in all five cases, and the enhancement pattern was compared with CT findings in three cases. In three cases, the cavernous sinus was involved unilaterally, with adjacent extracranial infiltrative masses. In one case, both orbits, the cavernous sinuses, and the tentorium were involved with diffuse infiltrative lesions. One patient had an infiltrative nasopharyngeal mass; and in all five patients, MR images showed localized involvement of the skull base, with bone marrow replaced by tumor. The soft-tissue lesions were hypointense on T2-weighted images in all five cases and showed homogeneous contrast enhancement. Histopathologic studies revealed scanty inflammatory cell infiltration with densely fibrotic background in all cases. The hypointensity of the lesions on T2-weighted images seemed to be related to the degree of fibrosis. Fibrosing inflammatory pseudotumor shows characteristic MR findings of infiltrative lesion with bone destruction and hypointensity on T2-weighted images. The lack of mobile protons due to the fibrotic background and/or high cellularity of the lesions may be the reason for their hypointensity and weaker enhancement on MR images.


Comparative targeted metabolomic profiles of porcine plasma and serum.
Metabolomics has been used to characterise many biological matrices and obtain detailed pictures of biological systems based on many metabolites. Plasma and serum are two blood-derived biofluids commonly used to assess and monitor the organismal metabolism and obtain information on the physiological and health conditions of an animal. Plasma is the supernatant that is separated from the cellular components after centrifugation of the blood that is first added with an anticoagulant. Serum is obtained after centrifugation of the blood that has been coagulated. The choice of one or the other biofluid for metabolomic analyses is related to specific analytical needs and technical issues, to problems derived by the collection and preparation steps, in particular when specimens are sampled from animals involved in field studies. Thus far, most of the metabolomic studies that compared plasma and serum have been carried out in humans and very little is known on the pigs. In this study, we used a targeted metabolomic platform that can detect about 180 metabolites of five biochemical classes to compare plasma and serum profiles of samples collected from 24 pigs. To also obtain a cross-species comparative metabolomic analysis, information for human plasma and serum derived from the same platform was retrieved from previous studies. Statistical analyses included univariate and multivariate approaches aimed at identifying stable and/or differentially abundant metabolites between the two porcine biofluids. A total of 154 (∼83%) metabolites passed the initial quality control, indicating a good repeatability of the analytical platform in pigs. Discarded metabolites included aspartate and biogenic amines that were already reported to be unstable in human studies. More than 80% of the metabolites had similar profiles in both porcine biofluids (average correlation was 0.75). Concentrations were usually higher in serum than in plasma, in agreement with what was already reported in humans. The univariate analysis identified 44 metabolites that had statistically different concentrations between porcine plasma and serum, of which 28 metabolites were also confirmed by the multivariate analysis. The obtained picture described similarities and differences between these two biofluids in pigs and the related human-pig comparisons. The obtained information can be useful for the choice of one or the other matrix for the implementation of metabolomic studies in this livestock species. The results can also provide useful hints to valuing the pig as animal model, in particular when metabolite-derived physiological states are relevant.


[Validation study on a multi-residue method for determination of pesticide residues in agricultural products by new automatic pretreatment equipment (FASRAC) and GC-MS/MS].
A validation study was performed on a multiresidue method for determination of pesticide residues in agricultural products according to the method validation guideline of the Ministry of Health, Labour and Welfare of Japan. FASRAC (Food Automatic Analytical Systems for Residual Agricultural Chemicals) automatically performs extraction of pesticide residues from agricultural products with acetonitrile, filtration, constant volume, mixing with the use of air, mixing acetonitrile with buffer solvent, separation, and dehydration with sodium sulfate. The extract was purified with a GC/NH2 column. For wheat flour and soybeans, a purification step with a C18 column was added before a GC/NH2 column. After removal of the solvent, the extract was resolved in n-hexane/acetone solvent for GC-MS/MS analysis. In the case of manual analysis, pesticide residues were analyzed according to official multiresidue methods and purification steps were the same as in FASRAC. Recovery tests were performed with wheat flour, soybeans, spinach and apples, by addition of 302 pesticides at the concentrations 0.01 mg/kg. The results indicate that automatic extraction using FASRAC is superior to manual analysis in trueness, repeatability and within-run reproducibility. Specially, automatic extraction using FASRAC is superior to manual analysis in trueness because it is optimized in various respects, for example reextraction at salting-out.


Danish very-low-dose aspirin after carotid endarterectomy trial.
The effect of very-low-dose aspirin as an antithrombotic agent was evaluated blindly in 301 patients who had recently undergone carotid endarterectomy. After randomization, 150 patients received aspirin and 151 received placebo. The two groups were comparable with regard to age, sex, blood pressure, previous cerebrovascular events, and smoking habits. The effect of the study medication on platelet aggregation was measured twice in each patient during the first 2 months and at each follow-up visit; the dose was individually adjusted. In 76% of the patients receiving aspirin, 50 mg/day gave satisfactory platelet inhibition, 13% needed 60 mg/day, 8% needed 70 mg/day, and 3% needed 100 mg/day. Platelet aggregation was found to be inhibited in only 1.2% of the measurements in the patients receiving placebo. Observation during treatment averaged 21 months; total intention-to-treat follow-up averaged 25 months. For the combined outcome events of transient ischemic attack, stroke, acute myocardial infarction, and vascular death, aspirin reduced risk by 11% (95% confidence limits: -38% to 48%, p greater than 0.1). Thus, there was no significant effect of very-low-dose aspirin in our trial.


First report of Onchocerca lupi from Israel and confirmation of two genotypes circulating among canine, feline and human hosts.
Onchocerca lupi is a parasitic filarioid and the causative agent of canine ocular onchocercosis, a zoonotic disease of domestic dogs with sporadic reports in humans. A 13-year-old dog with no travel history outside of Israel was presented to an ophthalmology veterinary clinic in Israel with severe right ocular and periocular disease. After surgical exploration, thin helminths were removed from the dorsal sclera of the eye and identified as Onchocerca lupi by polymerase chain reaction according to the cytochrome c oxidase subunit I (cox1), reduced nicotinamide adenine dinucleotide dehydrogenase subunit 5 (nad5) and 12S rRNA genes. Phylogenetic trees and haplotype networks of the cox1 and nad5 genes confirmed the circulation of two genotypes: genotype 1 with worms from dogs, cats and humans from both the Old and New Worlds, and genotype 2 with specimens from Portugal and Spain. The Israeli sequences clustered in genotype 1 and were identical to O. lupi from the USA. Evidence of two genotypes separated geographically sheds light on the phylogeography and evolution of this zoonotic pathogen, and suggests a diverse pathology observed in different regions of the world.


Not sold here: limited access to legally available syringes at pharmacies in Tijuana, Mexico.
Sterile syringe access is a critical component of HIV prevention programs. Although retail pharmacies provide convenient outlets for syringe access, injection drug users (IDUs) may encounter barriers to syringe purchase even where purchase without a prescription is legal. We sought to obtain an objective measure of syringe access in Tijuana, Mexico, where IDUs report being denied or overcharged for syringes at pharmacies. Trained "mystery shoppers" attempted to buy a 1 cc insulin syringe according to a predetermined script at all retail pharmacies in three Tijuana neighborhoods. The same pharmacies were surveyed by telephone regarding their syringe sales policies. Data on purchase attempts were analyzed using basic statistics to obtain an objective measure of syringe access and compared with data on stated sales policies to ascertain consistency. Only 46 (28.4%) of 162 syringe purchase attempts were successful. Leading reasons for unsuccessful attempts were being told that the pharmacy didn't sell syringes (35.3%), there were no syringes in stock (31.0%), or a prescription was required (20.7%). Of 136 pharmacies also surveyed by telephone, a majority (88.2%) reported selling syringes but only one-third (32.5%) had a successful mystery shopper purchase; the majority of unsuccessful purchases were attributed to being told the pharmacy didn't sell syringes. There was similar discordance regarding prescription policies: 74 pharmacies said in the telephone survey that they did not require a prescription for syringes, yet 10 of these pharmacies asked the mystery shopper for a prescription. IDUs in Tijuana have limited access to syringes through retail pharmacies and policies and practices regarding syringe sales are inconsistent. Reasons for these restrictive and inconsistent practices must be identified and addressed to expand syringe access, reduce syringe sharing and prevent HIV transmission.


Effect of steam explosion on physicochemical properties of waste activated sludge and the performance of anaerobic digestion.
The effect of steam explosion on physicochemical properties of sludge and the performance of anaerobic digestion (AD) was investigated. The steam explosion was conducted under different combinations of temperature and time, ranging 151-198 °C and 4-12 min respectively. The capillary suction time (CST) and viscosity of the sludge was increased with particle size decreased by improved hydrolysis temperature and prolonged hydrolysis time. The best sludge solubilization achieved was 41.3% under pretreatment condition of 198 °C and 4 min. Biogas production was enhanced with the improved sludge solubilization, and a linear correlation was found between biogas production and the severity factor (logR0) of steam explosion. However, the biogas productivity was reduced when the logR0 was increased from 3.79 to 3.96, probably owing to the generation of refractory organics during the high severity pretreatment. The temperature of 198 °C and the time of 8 min were the recommended operation parameters of steam explosion pretreatment for sludge AD, which could improve biogas production by 99.7 mL/g VSfed. The pH and NH4+-N during sludge AD was increased by steam explosion pretreatment; however, no inhibition on biogas production was observed.


Lifestyle factors associated with a rapid decline in the estimated glomerular filtration rate over two years in older adults with type 2 diabetes-Evidence from a large national database in Japan.
The present study investigated lifestyle risk factors from metabolic syndrome-related lifestyles for a rapid decline in the estimated glomerular filtration rate (eGFR) among adults aged 40-74 years with treated and untreated type 2 diabetes. This study analyzed data from the National Database of Health Insurance Claims and Specific Health Checkups of Japan, encompassing the period from fiscal year (FY) 2017 to FY2020. We established FY2018 as our baseline year. The subjects of this study were adults aged 40-74 in FY2018(baseline) who had type 2 diabetes and underwent specific health checkups in FY2020. We excluded adults with a medical record of kidney dialysis between FY2017 and FY2018, records of suspected type 1 diabetes between FY2017 and FY2020, or a baseline eGFR >85 mL/min/1.73 m2 or missing eGFR data as of FY2020. Eventually we analyzed 573,860 individuals. The outcome variable was a rapid decline in eGFR (≥30%) during the follow-up. Exposure lifestyle factors included skipping breakfast, late-night dinners, regular smoking, a high alcohol intake, non-refreshing sleep, and a lack of habitual exercise. Logistic regression models were stratified by age (40-59 and 60-74 years) and baseline eGFR levels (60-85, 30-59, and <30 mL/min/1.73 m2). Covariates included sex, a history of heart disease, a history of stroke, a history of renal failure, anemia, low-density lipoprotein, systolic blood pressure, hemoglobin A1C, body mass index, antidiabetic medications, antihypertension drugs, lipid-lowering drugs, the oral adsorbent Kremezin, non-steroidal anti-inflammatory drugs, and drugs for the treatment of renal anemia. A rapid decline in eGFR was detected in approximately 1.3% of participants (7,683 cases). In the baseline eGFR >30 subgroups (60-85 or 30-59 mL/min/1.73 m2), skipping breakfast and regular smoking were associated with a rapid decline in eGFR in both age groups, while a lack of habitual exercise and late-night dinners in the 60-74 age group and non-refreshing sleep in the 40-59 age group were identified as risk factors. Additionally, skipping breakfast was a risk factor for a rapid decline in eGFR in the 60-74 age group regardless of baseline eGFR levels. In the baseline eGFR <30 mL/min/1.73 m2 subgroup, skipping breakfast and non-refreshed sleep were risk factors for a rapid decline in eGFR. We found specific lifestyle risk factors were associated with a rapid eGFR decline among people with type 2 diabetes from a nationwide database in Japan. The associations varied by baseline eGFR level, age, and sex. Lifestyle modifications may effectively prevent the aggravation of diabetic kidney disease.


National Estimates of Home Care Workers Nonfatal Emergency Department-Treated Injuries, United States 2015-2020.
Home care workers are a critical resource contributing to the well-being of others. Presented are data on home care workers (HCWs) nonfatal emergency department-treated (ED) injuries. Nonfatal injuries among HCWs were extracted from the NEISS-Work data between 2015 and 2020. Review of NEISS-Work data indicated 117,000 HCWs with nonfatal ED-treated injuries; female HCWs accounted for 93%. Overexertion and bodily reactions accounted for 52% of the injuries. Violence and other injuries by persons or animals accounted for 15% and falls, slips, and trips also accounted for 15% of the HCWs ED-treated injuries. The growing demand for home care services is increasing the number of workers at risk for injury. Future analyses should prioritize injury events among HCWS to gain a better understanding of the events contributing to injuries among HCWs.


A case of lipoma of the deep lobe of the parotid gland.
Lipoma of the deep lobe of the parotid gland is extremely rare. A forty-two-year-old man presented with a painless, soft, and slow-growing mass in the left preauricular area. Ultrasonography and computed tomography findings were consistent with lipoma. The patient was treated with superficial parotidectomy and total excision of the mass with preservation of the facial nerve. Histologic findings confirmed the diagnosis. No recurrences were detected during 1.5 years in the postoperative period.


Genome Wide Mapping of Foxo1 Binding-sites in Murine T Lymphocytes.
The Forkhead box O (Foxo) family of transcription factors has a critical role in controlling the development, differentiation, and function of T cells. However, the direct target genes of Foxo transcription factors in T cells have not been well characterized. In this study, we focused on mapping the genome wide Foxo1-binding sites in naïve CD4+ T cells, CD8+ T cells, and Foxp3+ regulatory T (Treg) cells. By using chromatin immunoprecipitation coupled with deep sequencing (ChIP-Seq), we identified Foxo1 binding sites that were shared among or specific to the three T cell populations. Here we describe the experiments, quality controls, as well as the deep sequencing data. Part of the data analysis has been published by Ouyang W et al. in Nature 2012[1] and Kim MV et al. in Immunity 2013[2], and the associated data set were uploaded to NCBI Gene Expression Omnibus.


Fundamental photophysics of isomorphic and expanded fluorescent nucleoside analogues.
Fluorescent nucleoside analogues (FNAs) are structurally diverse mimics of the natural essentially non-fluorescent nucleosides which have found numerous applications in probing the structure and dynamics of nucleic acids as well as their interactions with various biomolecules. In order to minimize disturbance in the labelled nucleic acid sequences, the FNA chromophoric groups should resemble the natural nucleobases in size and hydrogen-bonding patterns. Isomorphic and expanded FNAs are the two groups that best meet the criteria of non-perturbing fluorescent labels for DNA and RNA. Significant progress has been made over the past decades in understanding the fundamental photophysics that governs the spectroscopic and environmentally sensitive properties of these FNAs. Herein, we review recent advances in the spectroscopic and computational studies of selected isomorphic and expanded FNAs. We also show how this information can be used as a rational basis to design new FNAs, select appropriate sequences for optimal spectroscopic response and interpret fluorescence data in FNA applications.


Autoinhibition mechanism of proto-Dbl.
The dbl oncogene encodes a prototype member of the Rho GTPase guanine nucleotide exchange factor (GEF) family. Oncogenic activation of proto-Dbl occurs through truncation of the N-terminal 497 residues. The C-terminal half of proto-Dbl includes residues 498 to 680 and 710 to 815, which fold into the Dbl homology (DH) domain and the pleckstrin homology (PH) domain, respectively, both of which are essential for cell transformation via the Rho GEF activity or cytoskeletal targeting function. Here we have investigated the mechanism of the apparent negative regulation of proto-Dbl imposed by the N-terminal sequences. Deletion of the N-terminal 285 or C-terminal 100 residues of proto-Dbl did not significantly affect either its transforming activity or GEF activity, while removal of the N-terminal 348 amino acids resulted in a significant increase in both transformation and GEF potential. Proto-Dbl displayed a mostly perinuclear distribution pattern, similar to a polypeptide derived from its N-terminal sequences, whereas onco-Dbl colocalized with actin stress fibers, like the PH domain. Coexpression of the N-terminal 482 residues with onco-Dbl resulted in disruption of its cytoskeletal localization and led to inhibition of onco-Dbl transforming activity. The apparent interference with the DH and PH functions by the N-terminal sequences can be rationalized by the observation that the N-terminal 482 residues or a fragment containing residues 286 to 482 binds specifically to the PH domain, limiting the access of Rho GTPases to the catalytic DH domain and masking the intracellular targeting function of the PH domain. Taken together, our findings unveiled an autoinhibitory mode of regulation of proto-Dbl that is mediated by the intramolecular interaction between its N-terminal sequences and PH domain, directly impacting both the GEF function and intracellular distribution.


Monthly variation in pediatric visits for headache to US emergency departments.
Headache is a common pediatric symptom often associated with stress and fatigue which may be more common during the school year. The purpose of this study is to determine if visits for headache are more common during the months of the school year. This study is a secondary analysis of the National Hospital Ambulatory Medical Care Survey (NHAMCS) database from 2001 to 2010. Patient visits in those aged 18 years or younger not associated with injury were examined. Only visits with an International Classification of Diseases, ninth revision (ICD-9) code consistent with headache were included. Data were analyzed using cumulative binomial probabilities. This statistic was used to establish the chance of seeing up to the observed number of visits for headache in a given month assuming that all months have an equal number of visits. A total of 660 unweighted visits representing 3.2 million patient encounters met the inclusion and exclusion criteria. Visits for headache were more common during the months of January, September, and October and less common in March, April, July, and November. Subgroup analysis was performed for children aged 13-18 years. In this subgroup, headaches were more common in January, September, and October. They were less common in July and December. Headache is more common during the first 2 months back to school in the fall as well as after the winter break in January. While we are not able to establish causality, we propose that children with headache require additional attention during the school year, particularly in the months following summer and winter breaks.


Concentrations of triglycerides, free fatty acids and glycerol in cord blood of newborn infants with a birth weight of less than or equal to 2700 grams.
Concentrations of triglycerides, free fatty acids (FFA) and glycerol were measured in umbilical venous blood from 99 infants with a birth weight of between 1100-2700 g and a gestational age of 27-41 weeks. Thirty infants were small for gestational age (SGA), 58 were appropriate (AGA) and 11 were of uncertain gestational age. In AGA infants with a gestational age of less than or equal to 35 weeks. FFA values were lower than in those with a gestational age of less than 35 weeks; otherwise concentrations of triglycerides, FFA and glycerol were independent of birth weight and gestational age in AGA infants. In SGA infants, higher FFA values were found compared with both AGA and term infants of normal birth weight. Triglyceride values were higher in SGA than in AGA infants. In SGA infants, a significant positive correlation was found between gestational age and concentrations of both FFA and triglycerides. No differences in FFA, glycerol and triglyceride concentrations were seen between asphyxiated and non-asphyxiated AGA infants.


Expression of G alpha i2 mimics several aspects of LPS priming in a murine macrophage-like cell line.
Priming of macrophages with low concentrations of lipopolysaccharide (LPS) enhances the ability of substances that act through heterotrimeric G proteins to stimulate immune cell functions. Although LPS-induced alterations in the expression and functions of G proteins of the alpha i family have been reported in hematopoietic cells, their effects on subsequent steps in LPS priming of macrophages have not been defined. To study the role of G alpha i2 in priming of macrophages by LPS, we expressed a mutant, activated form of alpha i2 (alpha i2Q205L) in P388D1 cells, and compared its effects on PAF-dependent C alpha signalling and arachidonic acid release to those in cells treated with LPS. In control P388D1 cells, treatment with LPS (100 ng/ml) for 1 hr increased the amount of alpha i2 protein 2-fold. Both LPS treatment and expression of alpha i2Q205L increased the rate of PAF-induced C alpha influx across the cell membrane and arachidonic acid release, although neither altered release of C alpha from intracellular stores by PAF. Expression of alpha i2Q205L is sufficient to mimic the effects of LPS on the PAF-induced C alpha i signal and enhanced arachidonic acid release. Consequently, although increasing the expression of alpha i2 may not be the sole mechanism by which LPS enhances signalling by PAF, increased alpha i2 expression can account for the alterations in PAF-induced C alpha i regulation, and arachidonic acid release in LPS-primed P388D1 cells.