Crystal structure of SARS-CoV-2 Orf9b in complex with human TOM70 suggests unusual virus-host interactions.
Although the accessory proteins are considered non-essential for coronavirus replication, accumulating evidences demonstrate they are critical to virus-host interaction and pathogenesis. Orf9b is a unique accessory protein of SARS-CoV-2 and SARS-CoV. It is implicated in immune evasion by targeting mitochondria, where it associates with the versatile adapter TOM70. Here, we determined the crystal structure of SARS-CoV-2 orf9b in complex with the cytosolic segment of human TOM70 to 2.2 Å. A central portion of orf9b occupies the deep pocket in the TOM70 C-terminal domain (CTD) and adopts a helical conformation strikingly different from the β-sheet-rich structure of the orf9b homodimer. Interactions between orf9b and TOM70 CTD are primarily hydrophobic and distinct from the electrostatic interaction between the heat shock protein 90 (Hsp90) EEVD motif and the TOM70 N-terminal domain (NTD). Using isothermal titration calorimetry (ITC), we demonstrated that the orf9b dimer does not bind TOM70, but a synthetic peptide harboring a segment of orf9b (denoted C-peptide) binds TOM70 with nanomolar KD. While the interaction between C-peptide and TOM70 CTD is an endothermic process, the interaction between Hsp90 EEVD and TOM70 NTD is exothermic, which underscores the distinct binding mechanisms at NTD and CTD pockets. Strikingly, the binding affinity of Hsp90 EEVD motif to TOM70 NTD is reduced by ~29-fold when orf9b occupies the pocket of TOM70 CTD, supporting the hypothesis that orf9b allosterically inhibits the Hsp90/TOM70 interaction. Our findings shed light on the mechanism underlying SARS-CoV-2 orf9b mediated suppression of interferon responses.


Estimation of the Continuous and Discontinuous Leverage Effects.
This paper examines the leverage effect, or the generally negative covariation between asset returns and their changes in volatility, under a general setup that allows the log-price and volatility processes to be Itô semimartingales. We decompose the leverage effect into continuous and discontinuous parts and develop statistical methods to estimate them. We establish the asymptotic properties of these estimators. We also extend our methods and results (for the continuous leverage) to the situation where there is market microstructure noise in the observed returns. We show in Monte Carlo simulations that our estimators have good finite sample performance. When applying our methods to real data, our empirical results provide convincing evidence of the presence of the two leverage effects, especially the discontinuous one.


[Results of provisional use of a system for voluntary anonymous reporting of incidents that threaten patient safety in the emergency medical services of Asturias].
To describe the reported incidents and adverse events in the emergency medical services of Asturias, Spain, and assess their consequences, delays caused, and preventability. Prospective, observational study of incidents reported by the staff of the emergency medical services of Asturias after implementation of a system devised by the researchers. Incident reports were received for 0.48% (95% CI, 0.41%-0.54%) of the emergencies attended. Patient safety was compromised in 74.7% of the reported incidents. Problems arising in the emergency response coordination center (ERCC) accounted for 37.6% of the incidents, transport problems for 13.4%, vehicular problems for 10.8%, and communication problems for 8.8%. Seventy percent of the reported incidents caused delays in care; 55% of the reported incidents that put patients at risk (according to severity assessment code ratings) corresponded to problems related to human or material resources. A total of 88.1% of the incidents reported were considered avoidable. Some type of intervention was required to attenuate the effects of 46.2% of the adverse events reported. The measures that staff members most often proposed to prevent adverse events were to increase human and material resources (28.3%), establish protocols (14.5%), and comply with quality of care recommendations (9.7%). It is important to promote a culture of safety and incident reporting among health care staff in Asturias given the number of serious adverse events. Reporting is necessary for understanding the errors made and taking steps to prevent them. The ERCC is the point in the system where incidents are particularly likely to appear and be noticed and reported.


Competitions and incentives for smoking cessation.
Material or financial incentives may be used in an attempt to reinforce behaviour change, including smoking cessation. They have been widely used in workplace smoking cessation programmes, and to a lesser extent within community programmes. Quit and Win contests are the subject of a companion review. To determine whether competitions and incentives lead to higher long-term quit rates. We also set out to examine the relationship between incentives and participation rates. We searched the Cochrane Tobacco Addiction Group Specialized Register, with additional searches of MEDLINE (January 1966 to September 2004), EMBASE (1980 to 2004/8), CINAHL (1982 to 2004/8) and PsycINFO (1872 to 2004/6). Search terms included incentive*, competition*, contest*, reward*, prize*, contingent payment*, deposit contract*. We considered randomized controlled trials, allocating individuals, workplaces, groups within workplaces, or communities to experimental or control conditions. We also considered controlled studies with baseline and post-intervention measures. Data were extracted by one author and checked by the second. We contacted study authors for additional data where necessary. The main outcome measure was abstinence from smoking for at least six months from the start of the intervention. We used the most rigorous definition of abstinence in each trial, and biochemically validated rates where available. Where possible we performed meta-analysis using a generic inverse variance model, grouped by timed endpoints, but not pooled across the subgroups. Fifteen studies met our inclusion criteria. None of the studies demonstrated significantly higher quit rates for the incentives group than for the control group beyond the six-month assessment. There was no clear evidence that participants who committed their own money to the programme did better than those who did not, or that different types of incentives were more or less effective. There is some evidence that although cessation rates have not been shown to differ significantly, recruitment rates can be improved by rewarding participation, which may be expected to deliver higher absolute numbers of successful quitters. Cost effectiveness analysis is not appropriate to this review, since the efficacy of the intervention has not been demonstrated. Incentives and competitions do not appear to enhance long-term cessation rates, with early success tending to dissipate when the rewards are no longer offered. Rewarding participation and compliance in contests and cessation programmes may have more potential to deliver higher absolute numbers of quitters.


Complexation, thermal and catalytic studies of N-substituted piperazine, morpholine and thiomorpholine with some metal ions.
Several Cu(II), Pt(II) and Ni(II) complexes of N-substituted, piperazine (NN donor), morpholine (NO donor) and thiomorpholine (NS donor) derivatives were synthesized and their thermal behavior and catalytic activity in epoxidation reaction of cis-diphenylethylene were studied using oxygen sources NaOCl. The coordination compounds of Cu(II), Pt(II) and Ni(II) having general formula [MLCl]Cl, [ML2l]Cl2 or [ML]Cl2 with tetra coordinated geometry around metal ions have been isolated as solid. All the ligands and complexes were identified by spectroscopic methods and elemental analysis, magnetic measurements, electrical conductance and thermal analysis. A square planer structures have been proposed for all complexes. The thermal stability of the complexes discussed in terms of ligands donor atoms, geometry and central metal ions. The complexes have a similar thermal behavior for the selected metal ions. The thermogravimetric analyses suggest high thermal stability for most complexes followed by thermal decomposition in different steps. The decomposition processes were observed as water elimination, chloride anion removal and degradation of the organic ligands. Catalytic ability of the complexes were examined and found that all the complexes can effectively catalyze the epoxidation of cis-stilbene with NaOCl.


Lung cancer in ferrous foundry workers: a review.
Epidemiologic studies indicate that an increased incidence of lung cancer may be associated with specific work areas in ferrous foundries. With the exception of crane operators, who were found to have an elevated lung cancer rate in one foundry, the excess lung cancer incidence is generally confined to molders, casters, and cleaning room operators whose lung cancer risk is two- to threefold higher than that of standard populations. These studies reflect conditions that existed in the foundries several decades ago. The lung cancer risk today may differ as a result of the introduction of new foundry practices and the use of new molding materials. Benzo(a)pyrene and certain other polycyclic aromatic hydrocarbons have been found in many locations in the foundries. It is not known if tumor promoters, co-carcinogens, or other classes of chemical carcinogens are present. The contribution of tobacco smoke to the lung cancer risk of ferrous foundry workers is also unknown. Current studies are examining the composition and long-term health effects of emissions from molds composed of modern synthetic chemical molding materials as well as those from the more traditional green sand molds.


[The "vanishing twin"].
Three cases of a vanishing twin are reported. We observed twin pregnancies in three women, who had their ultrasound examination before the 10th week of pregnancy. Positive heartbeat was registered in both embryos. When a further sonographic examination took place at the 11th week, intrauterine death of one of the embryos was observed. All three pregnancies continued as singleton pregnancies to uneventful term deliveries. When the placentae and chorionic membranes were examined after delivery, small nodular pigmented areas were observed. Histological examination showed the remains of the embryo, who had vanished in the first trimester of pregnancy.


A1 adenosine receptor antagonism improves glucose tolerance in Zucker rats.
The A1 adenosine receptor (A1ar) antagonist 1,3-dipropyl-8-(p-acrylic)-phenylxanthine (BW-1433) was administered to lean and obese Zucker rats to probe the influence of endogenously activated A1ars on whole body energy metabolism. The drug induced a transient increase in lipolysis as indicated by a rise in serum glycerol in obese rats. The disappearance of the response by day 7 of chronic studies was accompanied by an increase in A1ar numbers. Glucose tolerance tests were administered to rats treated with BW-1433. Peak serum insulin levels and areas under glucose curves (AUGs) were 34 and 41% lower in treated obese animals than in controls, respectively, and 19 and 39% lower in lean animals. With chronic administration (6 wk), AUGs decreased 47 and 33% in obese and lean animals, respectively. There was no effect of BW-1433 in either lean or obese rats on weight gain or percent body fat. Thus the major sustained influence of whole body A1ar antagonism in both lean and obese animals was an increase in whole body glucose tolerance at lower levels of insulin.


Dietary Nutrients and Cardiovascular Risk Factors among Renal Transplant Recipients.
Cardiovascular disease (CVD) is the leading cause of mortality in post-renal transplant recipients (RTRs). Adequate nutrient intake is a protective factor for CVD. We examined the associations of macronutrients and micronutrients with traditional and nontraditional CVD risk factors. Conducted from September 2016 to June 2018, this cross-sectional study included 106 RTRs aged ≥18 years with a functioning allograft. Dietary intake data from 3-day dietary records were collected. Nutrient intake adequacy was defined using various instruments, including the National Kidney Foundation Kidney Disease Outcomes Quality Initiative (K/DOQI) guidelines. CVD risk factors were defined according to the K/DOQI guidelines. Bivariate and multivariate logistic regression models were used to analyze the associations. CVD risk was present in all patients; the lowest proportions of adequate intake were 2.8% for dietary fiber and 0.9% for calcium. Adequate nutrient intake was associated with a lower likelihood of the occurrence of traditional CVD risk factors (specifically, 1.9-31.3% for hyperlipidemia and 94.6% for diabetes mellitus). It was also associated with a lower likelihood of the occurrence of nontraditional CVD risk by 0.8% for hypophosphatemia and 34% for hyperuricemia. Adherence to dietary guidelines should be promoted among RTRs to decrease CVD risk.


Evaluation of a ventral and a left lateral approach to coelioscopy in bearded dragons (Pogona vitticeps).
To compare a ventral and a left lateral endoscopic approach to coelioscopy in bearded dragons (Pogona vitticeps). 18 adult bearded dragons. In a randomized crossover design involving 2 surgical approaches, anesthetized bearded dragons first underwent coelioscopy with a ventral approach (left lateral of midline next to the umbilicus; animal positioned in dorsal recumbency) or left lateral approach (intercostal; animal positioned in right lateral recumbency) and then with the alternate approach. A 2.7-mm × 18-cm, 30° oblique telescope with a 4.8-mm operating sheath and CO2 insufflation at 2 to 5 mm Hg were used. Ease of entry into the coelom and ease of visual examination of visceral structures were scored. Both approaches were straightforward, with the left lateral approach requiring significantly more time than the ventral approach. Scores for ease of visual examination for the heart, lungs, liver, stomach, intestines, pancreas, gallbladder, left kidney, gonads, and fat body were good to excellent. Visual examination of the spleen and adrenal glands was difficult in most animals via either approach. The left kidney, testis, and vas deferens were easier to see with the left lateral approach, whereas the pancreas in females and gallbladder in both sexes were easier to see with the ventral approach. All bearded dragons recovered without complications from the procedures, except for one with nephritis, renal gout, and hepatic necrosis. Both coelioscopy approaches could be safely and effectively used in bearded dragons. Choice of approach should be based on the coelomic structures requiring evaluation.


Assessing students' self-reflective thinking in the classroom: the self-reflective thinking questionnaire.
The development of a questionnaire to assess students' use of self-reflective thinking in the classroom is described. On the basis of a literature search, items were selected. The items are students' self-report measures and open-ended questions. The participants were 96 fourth grade secondary vocational students from six classes in The Netherlands, all of whom were used to learning in cooperative groups. Complementary data were selected to validate this questionnaire. Visual inspection of the frequencies indicated a difference between levels of students' self-reflecting thinking. Between-subjects t tests showed that students' motivational engagement and marks could be used to validate the measure of self-reflective thinking. The implication of the questionnaire to assess students' self-reflective thinking within the classroom are discussed.


Seasonal suicide trends in adolescents in the US: Did they explain the apparent effect of the Netflix show 13 Reasons Why?
Controversy surrounds the effects of the first season of the Netflix show 13 Reasons Why on adolescent suicide in the United States. This research reexamined the question using more sensitive weekly suicide counts and more sensitive tests for seasonal changes in suicide. Weekly suicide counts for adolescents ages 10-19 and young people ages 20-29 obtained from CDC were analyzed from 2013 to 2018 NVSS Mortality Data using time series methods. Adolescents ages 10-19 exhibited seasonal changes in suicides that increased from March to April and that also paralleled the school year. The same seasonal pattern was evident in males ages 20-24. There was no evidence of an effect of 13 Reasons in male adolescent suicides in 2017 once seasonality, auto-correlation, and longer term trends were removed. Seasonal changes and secular increases can explain why previous studies observed suicide increases in March and April of 2017 in male adolescents. The association between school year and suicide suggests that stressors associated with schooling increase the risk of suicide in some youth. How to reduce these risks is an important focus for future research as is the need to understand the conditions under which fictional depictions of suicide lead to contagion.


Patient characteristics, treatment patterns, and health outcomes among COPD phenotypes.
Recent literature has suggested that emphysema and chronic bronchitis, traditionally considered to be entities overlapping within chronic obstructive pulmonary disease (COPD), may be distinct disorders. Few studies have examined the differences in patient characteristics and health outcomes between these conditions. This study examined whether COPD phenotypes represent distinct patient populations, in a large nationally representative US sample. Data were obtained from the 2010 US National Health and Wellness Survey (NHWS). NHWS respondents (n = 75,000) were categorized as a COPD phenotype based on their self-reported diagnosis of COPD only (n = 970), emphysema only (n = 399), or chronic bronchitis only (n = 2071). Phenotypes were compared on demographics, health characteristics, treatment patterns, health outcomes, work productivity, and resource use. Variables were compared using Chi-square and analysis of variance tests for categorical and continuous outcomes, respectively. Health outcomes were also examined using regression modeling, controlling for demographic and health characteristic covariates. Patients with chronic bronchitis were significantly younger (51.38 years versus 63.24 years for COPD versus 63.30 years for emphysema, P < 0.05) and more likely to be employed (46.98% versus 23.81% for COPD versus 28.33% for emphysema, P < 0.05). Relative to the other phenotypes, patients with chronic bronchitis were also significantly more likely to be female, nonwhite, and to exercise currently (all P < 0.05), and were significantly less likely to be a current or former smoker (P < 0.05). Controlling for demographic and health characteristics, patients self-identified as having COPD only reported significantly worse physical quality of life (adjusted mean 36.69) and health utilities (adjusted mean 0.65) and significantly more absenteeism (adjusted mean 7.08%), presenteeism (adjusted mean 30.73%), overall work impairment (adjusted mean 34.06%), and activity impairment (adjusted mean 46.59%) than the other phenotypes (all P < 0.05). These results suggest considerable heterogeneity among different COPD phenotypes with respect to demographics, health characteristics, disease characteristics, treatment patterns, and health outcomes. Research aimed at understanding the differences in patient characteristics and disease presentation of these phenotypes could be used to guide treatment recommendations.


Whole blood resuscitation restores intestinal perfusion and influences gut microbiome diversity.
Gut dysbiosis, an imbalance in the gut microbiome, occurs after trauma, which may be ameliorated with transfusion. We hypothesized that gut hypoperfusion following trauma causes dysbiosis and that whole blood (WB) resuscitation mitigates these effects. Anesthetized rats underwent sham (S; laparotomy only, n = 6); multiple injuries (T; laparotomy, liver and skeletal muscle crush injuries, and femur fracture, n = 5); multiple injuries and 40% hemorrhage (H; n = 7); and multiple injuries, hemorrhage, and WB resuscitation (R; n = 7), which was given as 20% estimated blood volume from donor rats 1 hour posttrauma. Baseline cecal mesenteric tissue oxygen (O2) concentration was measured following laparotomy and at 1 hour and 2 hours posttrauma. Fecal samples were collected preinjury and at euthanasia (2 hours). 16S rRNA sequencing was performed on purified DNA, and diversity and phylogeny were analyzed with QIIME (Knight Lab, La Jolla, CA; Caporaso Lab, Flagstaff, AZ) using the Greengenes 16S rRNA database (operational taxonomic units; 97% similarity). α and β diversities were estimated using observed species metrics. Permutational analysis of variance was performed for overall significance. In H rats, an average decline of 36% ± 3.6% was seen in the mesenteric O2 concentration at 1 hour without improvement by 2 hours postinjury, which was reversed following resuscitation at 2 hours postinjury (4.1% ± 3.1% difference from baseline). There was no change in tissue O2 concentration in the S or T rats. β Diversity differed among groups for all measured indices except Bray-Curtis, with the spatial median of the S and R rats more similar compared with S and H rats (p < 0.05). While there was no difference in α diversity found among the groups, indices were significantly correlated with mesenteric O2 concentration. Members of the family Enterobacteriaceae were significantly enriched in only 2 hours. Mesenteric perfusion after trauma and hemorrhage is restored with WB resuscitation, which influences β diversity of the gut microbiome. Whole blood resuscitation may also mitigate the effects of hemorrhage on intestinal dysbiosis, thereby influencing outcomes.


Survival and drifting patterns of grass carp eggs and larvae in response to interactions with flow and sediment in a laboratory flume.
A series of laboratory experiments was conducted to better understand the behavior of grass carp eggs and larvae in moving water in order to develop and implement new strategies for control and prediction of their dispersal and drift at early life stages. Settling velocity and density of a representative sample of eggs were estimated, and three trials of flume experiments with different flow conditions were conducted with live eggs in a temperature-controlled setting with a mobile sediment bed. In these trials, egg and larval stages were continuously analyzed over periods of 80 hours; and eggs and larvae interactions with the flow and sediment bed were monitored and characterized qualitatively and quantitatively. Survival rates were quantified after each trial, highlighting physical causes for increased mortality. Detailed flow analysis was correlated to the observed drifting and swimming behavior of eggs and larvae, to estimate distributions across the water depth, as well as traveling and swimming speeds. Evidence of the influence of mean and turbulent flow in the suspension and transport of eggs are reported, and swimming patterns of larvae at different developmental stages are described. These findings support the development of new strategies for monitoring the spread of grass carp eggs and larvae in rivers, and provide new inputs to predict conditions favorable for spawning and hatching, allowing for mitigation measures at early life stages, which are critical to control their dispersal.


Ferulic acid protects lipopolysaccharide-induced acute kidney injury by suppressing inflammatory events and upregulating antioxidant defenses in Balb/c mice.
Sepsis-induced acute kidney injury (AKI) is responsible for 70-80% mortality in intensive care patients due to elevated levels of endotoxin, Lipopolysaccharide (LPS) caused by gram-negative infections. Ferulic acid (FA), a phenolic phytochemical is known for its renal protection on various induced models of nephrotoxicity. However, the curative effect of FA in LPS-induced AKI is not well studied. This study aimed to investigate the effect of FA on LPS-induced AKI in mice model and to understand the protective mechanisms involved, to provide evidence for FA in the treatment of AKI. Balb/c mice were treated with FA at 50 mg/kg and 100 mg/kg dosages after LPS stimulation (10 mg/kg). At the end of the intervention, we determined the concentrations of serum creatinine and blood urea nitrogen, inflammatory cytokines and histopathological changes in animals. Also, the relative protein expression level of TLR4 mediated NF-κB signaling pathway were studied in kidney tissues. FA treated animals showed upregulation of antioxidant defenses and suppression of inflammatory events by inhibiting TLR-4 mediated NFκB activation. However, LPS alone administered group, resulted in rapid renal damage with increased levels of blood urea nitrogen and modest increase in creatinine; decreased antioxidant defenses and release of inflammatory cytokines. The histopathological analysis also revealed the protective action of the FA against sepsis induced fibrosis and renal damage. Our findings demonstrated that FA exhibits marked protective effects on LPS-induced AKI in mice suggesting its chemopotential role for treating AKI in humans.


Hormonal alterations in heart failure: anabolic impairment in chronic heart failure - diagnostic, prognostic and therapeutic issues.
Chronic heart failure represents a leading cause of mortality and health care expenditure in developed countries. In the last 20 years, medical therapy of heart failure has dramatically changed thanks to the introduction of agents able to significantly reduce the neurohormonal hyperactivation that underpins the syndrome, and to the growing opportunities of electrical therapies. Although major advances in terms of improved survival and quality of life have been achieved, the reduction in the burden of heart failure is still the primary goal of cardiovascular societies. In the last decades, other research lines have also grown to complement the neurohormonal paradigm. It is increasingly evident that several hormonal systems are down-regulated or impaired in patients with heart failure, including growth and thyroid hormones, androgens and insulin. These abnormalities could be considered interrelated and linked, in turn, to the neurohormonal and cytokine hyperactivation. Since most of these alterations provide prognostic information, these new lines of evidence support the extension of the classical neurohormonal scheme to a more comprehensive pathophysiological model that includes multiple hormonal and metabolic deficiencies. This chapter examines the evidence in support of this concept. Preliminary experience concerning targeted hormonal supplementation or metabolic modulation is also briefly reviewed in this article.


Isotope effects in the transient phases of the reaction catalyzed by ethanolamine ammonia-lyase: determination of the number of exchangeable hydrogens in the enzyme-cofactor complex.
Transient phases of the reaction catalyzed by ethanolamine ammonia-lyase (EAL) from Salmonella typhimurium have been investigated by stopped-flow visible spectrophotometry and deuterium kinetic isotope effects. The cleavage of adenosylcobalamin (coenzyme B(12)) to form cob(II)alamin (B(12r)) with ethanolamine as the substrate occurred within the dead time of the instrument whenever coenzyme B(12) was preincubated with enzyme prior to mixing with substrate. The rate was, however, slowed sufficiently to be measured with perdeutero ethanolamine as the substrate. Optical spectra indicate that, during the steady states of the reactions with ethanolamine and with S-2-aminopropanol as substrates, approximately 90% of the active sites contain B(12r). Reformation of the carbon-cobalt bond of the cofactor occurs following depletion of substrate in the reaction mixtures, and the rate constant for this process reflects k(cat) of the respective substrates. This late phase of the reaction also exhibits (2)H isotope effects similar to those measured for the overall reaction with (2)H-labeled substrates. With unlabeled substrates, the rate of cofactor reassembly is independent of the number of substrate molecules turned over in the steady-state phase. However, with (2)H-labeled substrates, kinetic isotope effects appear in the reassembly phase, and these isotope effects are maximal after only approximately 2 equiv of substrate/active site are processed. With 5'-deuterated coenzyme B(12) and deuterated substrate, the isotope effect on reassembly is independent of the number of substrate molecules that are turned over. These results indicate that the pool of exchangeable hydrogens in the enzyme-cofactor complex is two-a finding consistent with the hydrogens in the C5' methylene of coenzyme B(12).


Zebrafish class 1 phosphatidylinositol transfer proteins: PITPbeta and double cone cell outer segment integrity in retina.
Phosphatidylinositol transfer proteins (PITPs) in yeast co-ordinate lipid metabolism with the activities of specific membrane trafficking pathways. The structurally unrelated metazoan PITPs (mPITPs), on the other hand, are an under-investigated class of proteins. It remains unclear what biological activities mPITPs discharge, and the mechanisms by which these proteins function are also not understood. The soluble class 1 mPITPs include the PITPalpha and PITPbeta isoforms. Of these, the beta-isoforms are particularly poorly characterized. Herein, we report the use of zebrafish as a model vertebrate for the study of class 1 mPITP biological function. Zebrafish express PITPalpha and PITPbeta-isoforms (Pitpna and Pitpnb, respectively) and a novel PITPbeta-like isoform (Pitpng). Pitpnb expression is particularly robust in double cone cells of the zebrafish retina. Morpholino-mediated protein knockdown experiments demonstrate Pitpnb activity is primarily required for biogenesis/maintenance of the double cone photoreceptor cell outer segments in the developing retina. By contrast, Pitpna activity is essential for successful navigation of early developmental programs. This study reports the initial description of the zebrafish class 1 mPITP family, and the first analysis of PITPbeta function in a vertebrate.


Heavy crisis, new perspectives? Investigating the role of consumption, time wealth and meaning construction during countrywide Covid-19 lockdown in Germany.
The COVID-19 pandemic profoundly changed the way we live and consume. One open question is whether the crisis provides an opportunity to increase alternatives to materialistic consumption. We characterize these alternatives as sustainable leisure behaviour. Our study aimed to analyse: (i) Changes in behaviour frequency of consumption and sustainable leisure before and during lockdown. (ii) How potential changes in behaviours are appraised and if there are intentions to keep changed behaviour. (iii) Influence of Time Wealth (an alternative model of affluence that potentially promotes low consumption life styles) and Life Reflection (fundamental reflection processes stimulated by the Covid-19 crisis) on consumption and sustainable leisure behaviour during lockdown. We collected data from 947 participants in Germany, using an online survey. Participants reported behaviour frequencies of consumption and sustainable leisure before and during lockdown. Furthermore, participants evaluated potential behaviour changes and rated statements regarding their future intentions. Main findings: (i) Pairwise t-tests revealed reduced consumption behaviour in Electronics and Clothes. All sustainable leisure behaviours increased during lockdown. (ii) Increases in sustainable behaviour received positive evaluation and were intended to extend into the future. Consumption behaviour results were mixed. (iii) In multiple regression analysis, Time Wealth and Life Reflection were positively related to most sustainable leisure behaviour. Sustainable leisure behaviour correlated positively with Life Satisfaction and Presence of Meaning. We discuss future research ideas regarding the promotion of sustainable wellbeing in a post COVID society.


Hepatocellular Carcinoma: An Uncommon Metastasis in the Orbit.
Hepatocellular carcinoma (HCC) represents the most common type of primary cancer of the liver and is associated with poor prognosis. It is the most common cause of death in cirrhotic patients and in different studies was shown as the third most common cause of cancer-related deaths worldwide. Each year, approximately half a million people are diagnosed with HCC. In recent decades, the prognosis of patients with HCC has improved because more cases are diagnosed and treated at early stages; high-risk patients (i.e., with chronic HBV or HCV infection) are followed more often for the possibility of HCC, and novel treatment options such as locoregional therapy are used with better overall results. The extrahepatic metastases represent a poor prognostic factor. The most common sites of metastasis in advanced hepatocellular carcinoma are the lung (44%), portal vein (35%), and portal lymph nodes (27%). Also, intra-abdominal lymph nodes and bones are common sites. Orbital metastases rarely occur, representing the 3-7% of orbital masses. These metastases are usually found in advanced tumor stages. The mechanism of metastasis to the orbit is difficult to determine. A hematogenous route, as for other primary neoplasms of the abdomen, may be suspected. Tumor cells may circulate through the vena cava, beyond the pulmonary filter to the heart, and finally be distributed to the orbital region through the arterial systemic circulation. We describe herein a case of an adult male with liver cirrhosis due to alcohol abuse who presented with concomitant diagnosis of HCC and orbit metastasis.


Cholesterol levels in untreated Spanish hypertensive patients. The Compas Study Group, Spanish Hypertension Society.
In daily practice, arterial hypertension (AHT) and hypercholesterolaemia are frequently associated with the existence of multiple common etiopathogenic interrelationships. This situation leads to an exponential increase in cardiovascular risk for these patients, so it is essential to know the prevalence and therapeutic management of hypercholesterolaemia in the hypertensive patient. This national study analyses the distribution of total cholesterol levels and low-density lipoprotein cholesterol as well as hypercholesterolaemia prevalence and its therapeutic management in the uncontrolled hypertensive Spanish population. We observed mean total cholesterol levels of 227+/-41 mg/dl with a high prevalence of hypercholesterolaemia (34.2%) among hypertensive patients, and the percentage of those patients with "desirable" total cholesterol levels (<200 mg/dl) was <25%. The treated hypertensive patients presented both significantly higher mean cholesterol levels and greater hypercholesterolaemia prevalence than the untreated hypertensive patients. It appears that total cholesterol levels are scarcely related to the presence or non-presence of obesity, diabetes or smoking. Regarding treatment, only 14.6% of the hypercholesterolaemic hypertensive patients received hypolipaemic treatment with statins. These results support the need to introduce measures for better diagnostic and therapeutic management of hypercholesterolaemic hypertensive patients that will lead to a much higher reduction in cardiovascular risk for these patients.


Psychometric properties and factor structure of the Behavior Problems Inventory (BPI-01) in a Swedish community population of adults with intellectual disability.
The aim of the present study was to evaluate the psychometric properties and factor structure of the Behavior Problems Inventory (BPI-01) in a community population. The Swedish version of the BPI-01 was administered by interviewing care staff of all adults (n = 915) with administratively defined intellectual disabilities (IDs) living in Örebro County, Sweden. Sixty-two percent of the participants had at least one behavior problem. Altogether, 30.9% showed self-injurious behavior, 41.3% stereotyped behavior, and 34.8% aggressive/destructive behavior. All but the self-injurious behavior scale reached acceptable levels of internal consistency. Confirmatory factor analysis supported the unidimensionality of the subscales as well as the proposed three factor structure of the original BPI-01. The present study demonstrates that the three subscales are highly similar constructs across different language and cultural settings, and that the BPI-01 is applicable in research on populations with varying mental functioning, diagnoses, ages, and living arrangements.


Herbal remedies as alternative to conventional therapies for the treatment of pediatric infectious diseases.
Pediatric infections still represent a leading cause of mortality in many developing countries. Since ancient times, traditional healing systems provided some herbal remedies to treat pediatric diseases, only in some cases validated by an evidence-based approach. Therefore, this review covers the herbal remedies in Iranian traditional medicine and aims to assess the potential of phytotherapeutics as safe and effective alternatives to conventional therapies for the treatment of pediatric infectious diseases. Notably, pediatric patients may also benefit from adjuvant therapy, i.e., combined treatment with herbal remedies and conventional therapies, to improve the efficacy of conventional drugs, decrease their adverse effects at the cell-tissue-organ-organism level and reduce the occurrence of microbial strains resistant to antibiotics. Therefore, traditional healing systems still represent an unlimited source of active ingredients to be tested in preclinical assays as well as in humans in terms of efficacy and safety.


β adrenergic receptor modulation of neurotransmission to cardiac vagal neurons in the nucleus ambiguus.
β-adrenergic receptors are a class of G protein-coupled receptors that have essential roles in regulating heart rate, blood pressure, and other cardiorespiratory functions. Although the role of β adrenergic receptors in the peripheral nervous system is well characterized, very little is known about their role in the central nervous system despite being localized in many brain regions involved in autonomic activity and regulation. Since parasympathetic activity to the heart is dominated by cardiac vagal neurons (CVNs) originating in the nucleus ambiguus (NA), β adrenergic receptors localized in the NA represent a potential target for modulating cardiac vagal activity and heart rate. This study tests the hypothesis that activation of β adrenergic receptors alters the membrane properties and synaptic neurotransmission to CVNs. CVNs were identified in brainstem slices, and membrane properties and synaptic events were recorded using the whole-cell voltage-clamp technique. The nonselective β agonist isoproterenol significantly decreased inhibitory GABAergic and glycinergic as well as excitatory glutamatergic neurotransmission to CVNs. In addition, the β(1)-selective receptor agonist dobutamine, but not β(2) or β(3) receptor agonists, significantly decreased inhibitory GABAergic and glycinergic and excitatory glutamatergic neurotransmission to CVNs. These decreases in neurotransmission to CVNs persisted in the presence of tetrodotoxin (TTX). These results provide a mechanism by which activation of adrenergic receptors in the brainstem can alter parasympathetic activity to the heart. Likely physiological roles for this adrenergic receptor activation are coordination of parasympathetic-sympathetic activity and β receptor-mediated increases in heart rate upon arousal.


Health care technology in the Federal Republic of Germany.
The basic goal of the German health care system is equal access to all medical services for all citizens. The federal government sets the legal framework for the system, but most health policy decisions are made through bargaining between large organizations within a legal framework. The federal government took little active role in health care until the early 1990s, when it became increasingly apparent that budgeting and other cost containment measures seemed to be insufficient to successfully reduce the growth of mandatory sickness funds expenditures. A 1993 law has attempted to address some of the most obvious deficiencies in the system, while encouraging a market-oriented approach to health care. Health care technology assessment has almost no role in the German health care system. Attempts by professionals and politicians to introduce technology assessment into the health care arena have been largely unsuccessful.


Structure of the full-length human RPA14/32 complex gives insights into the mechanism of DNA binding and complex formation.
Replication protein A (RPA) is the ubiquitous, eukaryotic single-stranded DNA (ssDNA) binding protein and is essential for DNA replication, recombination, and repair. Here, crystal structures of the soluble RPA heterodimer, composed of the RPA14 and RPA32 subunits, have been determined for the full-length protein in multiple crystal forms. In all crystals, the electron density for the N-terminal (residues 1-42) and C-terminal (residues 175-270) regions of RPA32 is weak and of poor quality indicating that these regions are disordered and/or assume multiple positions in the crystals. Hence, the RPA32 N terminus, that is hyperphosphorylated in a cell-cycle-dependent manner and in response to DNA damaging agents, appears to be inherently disordered in the unphosphorylated state. The C-terminal, winged helix-loop-helix, protein-protein interaction domain adopts several conformations perhaps to facilitate its interaction with various proteins. Although the ordered regions of RPA14/32 resemble the previously solved protease-resistant core crystal structure, the quaternary structures between the heterodimers are quite different. Thus, the four-helix bundle quaternary assembly noted in the original core structure is unlikely to be related to the quaternary structure of the intact heterotrimer. An organic ligand binding site between subunits RPA14 and RPA32 was identified to bind dioxane. Comparison of the ssDNA binding surfaces of RPA70 with RPA14/32 showed that the lower affinity of RPA14/32 can be attributed to a shallower binding crevice with reduced positive electrostatic charge.


Perturbation effects in dosimetry: Part I. Kilovoltage x-rays and electrons.
Perturbation effects are defined as departures from ideal large-detector or Bragg-Gray cavity behaviour. Such effects are central to the use of practical dosimeters for accurate dose determination, as is required in external-beam radiotherapy. A theoretical framework for treating perturbation effects is established. In this first part of the review, perturbation in kilovoltage x-ray and megavoltage electron beams are treated in detail, with the emphasis on ionization chambers. The displacement factor for ion chambers in kilovoltage x-ray beams is discussed, starting with the early, pioneering work of Lamerton and Lidén. The evidence for the large values of the perturbation factor in medium-energy x-ray beams (between 100 and 300 kV) recommended in the 1987 IAEA dosimetry code is critically examined and revised, smaller values are given. In electron beams the theoretical approaches to the correction for the in-scattering correction in gas-filled cavities is discussed in detail. The evidence for negligible perturbation in low-energy electron beams in plane-parallel chambers with adequate guardring widths is critically reviewed, including the suggested correction for perturbation due to backscattering differences between the chamber-wall material and the medium. The various models for the response of thermoluminescent dosimeters in electron beams are discussed. It is concluded that Monte Carlo simulation of dosimeter response is likely to play an even bigger role in the future.


Depressed prostaglandins and leukotrienes in veterans with Gulf War illness.
Background: There is need to understand biological markers and mechanisms in Gulf War illness (GWI). Goal: To examine whether and how eicosanoids - prostaglandins and leukotrienes - are altered in veterans with GWI. Methods: Seventy participants including 37 GWI and 33 healthy controls, shared exposure information, and had plasma eicosanoids assessed - prostaglandin F2 alpha (pgf2α), prostaglandin D2 (pgd2), leukotriene B4 (lb4) among others. Values were compared for GWI versus controls. Eicosanoid intercorrelations were compared in cases vs. controls. For the most significantly altered eicosanoid in GWI, exposure and symptom relations were assessed. Results: Prostaglandins and leukotrienes were depressed in GWI, strongest for pgf2α, then lb4. Eicosanoid intercorrelations differed in GWI vs. controls. Fuel-solvent, pesticide, radioactive chemicals and metal exposures related negatively to pgf2α; as, in GWI, did chemical attack and vaccines. Multivariate predictors included fuels-solvents and radioactive chemicals (negative); tetanus vaccine and herbicides (positive). Fuels-solvents and radioactive chemicals predicted lower pgf2α in cases, controls, and all participants controlled for case status. Lower pgf2α related to GWI "Kansas criteria" domains of pain, respiratory, and (borderline significantly) skin symptoms. Conclusion: Multiple eicosanoids are depressed in GWI, particularly pgf2α and lb4. Prior fuel-solvent exposures, radioactive chemicals, and (in GWI cases) vaccines were linked to lower pgf2α.


Primate Torpor: Regulation of Stress-activated Protein Kinases During Daily Torpor in the Gray Mouse Lemur, Microcebus murinus.
Very few selected species of primates are known to be capable of entering torpor. This exciting discovery means that the ability to enter a natural state of dormancy is an ancestral trait among primates and, in phylogenetic terms, is very close to the human lineage. To explore the regulatory mechanisms that underlie primate torpor, we analyzed signal transduction cascades to discover those involved in coordinating tissue responses during torpor. The responses of mitogen-activated protein kinase (MAPK) family members to primate torpor were compared in six organs of control (aroused) versus torpid gray mouse lemurs, Microcebus murinus. The proteins examined include extracellular signal-regulated kinases (ERKs), c-jun NH2-terminal kinases (JNKs), MAPK kinase (MEK), and p38, in addition to stress-related proteins p53 and heat shock protein 27 (HSP27). The activation of specific MAPK signal transduction pathways may provide a mechanism to regulate the expression of torpor-responsive genes or the regulation of selected downstream cellular processes. In response to torpor, each MAPK subfamily responded differently during torpor and each showed organ-specific patterns of response. For example, skeletal muscle displayed elevated relative phosphorylation of ERK1/2 during torpor. Interestingly, adipose tissues showed the highest degree of MAPK activation. Brown adipose tissue displayed an activation of ERK1/2 and p38, whereas white adipose tissue showed activation of ERK1/2, p38, MEK, and JNK during torpor. Importantly, both adipose tissues possess specialized functions that are critical for torpor, with brown adipose required for non-shivering thermogenesis and white adipose utilized as the primary source of lipid fuel for torpor. Overall, these data indicate crucial roles of MAPKs in the regulation of primate organs during torpor.


Primary anxiety disorders and the development of subsequent alcohol use disorders: a 4-year community study of adolescents and young adults.
Cross-sectional findings in community surveys of adults suggest that adolescent anxiety disorders are strong predictors of the subsequent onset of alcohol use, abuse and dependence. However, prospective data that follow a sample of adolescents into adulthood are needed to confirm these associations. Baseline and 4-year follow-up data from the EDSP-Study, a prospective community survey of 3021 (2548 at follow-up) adolescents and young adults aged 14 to 24 years at baseline carried out in Munich, were used. DSM-IV anxiety disorders, alcohol use and alcohol use disorders were assessed with the Munich-Composite-International-Diagnostic-Interview (M-CIDI). Multiple logistic regression analysis, controlling for age, gender, other mental disorders, substance use disorders and antisocial behaviour was used to study the associations of baseline anxiety disorders with the subsequent onset and course of alcohol use and alcohol disorders. Baseline social phobia significantly predicts the onsets of regular use and hazardous use and the persistence of dependence. Panic attacks significantly predict the onsets of hazardous use and abuse as well as the persistence of combined abuse/dependence. Panic disorder significantly predicts the persistence of combined abuse/dependence. Other anxiety disorders do not significantly predict any of the outcomes. Panic and social phobia are predictors of subsequent alcohol problems among adolescents and young adults. Further studies are needed to investigate the underlying mechanisms and the potential value of targeted early treatment of primary panic and social phobia to prevent secondary alcohol use disorders.


The role of reexcision for positive margins in optimizing local disease control after breast-conserving surgery for cancer.
One of the most important factors associated with local recurrence after lumpectomy in breast cancer patients is the status of the surgical margin. Standard surgical practice is to obtain clear margins even if this requires a second surgical procedure. It is assumed that reexcision to achieve clear margins when positive margins are present at initial excision is as effective as complete tumor removal at a single procedure; however, the efficacy of reexcision in this context has not been well studied. A retrospective search of the Henrietta Banting Breast Centre database from 1987 to 1997 identified 1430 patients who underwent lumpectomy for invasive breast cancer: 1225 patients (group A) had negative margins at the initial surgery and 152 patients (group B) underwent one or more reexcisions to achieve negative margins. Fifty-three patients had positive margins at final surgery, but no reexcision was done (group C). Logistic regression was used to identify factors that were predictive of a positive margin; predictors of local recurrence in women whose tumors were completely resected were determined using Cox's proportional hazards model. Patients in groups A, B, and C differed with respect to mean age at diagnosis (58 years, 51 versus, and 56 years, respectively, p < 0.0001), mean tumor size (19 mm, 16 mm, and 26 mm, respectively, p < 0.0001), node positivity (30%, 22%, and 41%, respectively, p = 0.004), and the presence of a ductal carcinoma in situ (DCIS) component (60%, 64%, and 79%, respectively, p = 0.007). The mean follow-up period was similar for the three groups (8 years, 8 years, and 9 years, respectively, p = 0.17). Young age was the only variable predictive of positive margins. Among patients undergoing complete tumor excision, there was a suggestion of a higher 10 year local recurrence rate in reexcision group B, but the difference did not reach statistical significance (11.6% versus 16.6%, p = 0.11). Cox's multivariate regression analyses identified older age, smaller tumor size, receiving radiation therapy, and tamoxifen use as significantly decreasing the rate of local recurrence in patients with negative margins at initial surgery or after reexcision. Our data confirm the results of previous studies indicating that young age is an independent predictor of positive margins after lumpectomy for invasive breast cancer. The only independent predictor of local recurrence in our study cohort was large tumor size. There was a trend toward a higher local recurrence rate if more than one procedure was required to secure clear margins, although this effect was not independent of other factors. Reexcision to clear involved margins is an important surgical intervention for both younger and older women.


Administering xCT Inhibitors Based on Circadian Clock Improves Antitumor Effects.
Clock genes encoding transcription factors that regulate circadian rhythms may inform chronomodulated chemotherapy, where time-dependent dose alterations might affect drug efficacy and reduce side effects. For example, inhibiting the essential cystine transporter xCT with sulfasalazine induces growth arrest in cancer cells. Although the anticancer effects of sulfasalazine have been studied extensively, its effects on transcriptional control of xCT expression have not been studied. Here, we show that sulfasalazine administration during the period of increased xCT expression improves its anticancer effects and that the Clock gene itself induces xCT expression and regulates its circadian rhythm. Our findings highlight the clinical potential of chronomodulated chemotherapy and the importance of xCT-mediated transcriptional regulation in the utility of such strategies. Cancer Res; 77(23); 6603-13. ©2017 AACR.


Growth-friendly surgery results in more growth but a higher complication rate and unplanned returns to the operating room compared to single fusion in neuromuscular early-onset scoliosis: a multicenter retrospective cohort study.
Compare radiographic outcomes, complications, and QoL in neuromuscular early-onset scoliosis (EOS) patients treated with single posterior spinal fusion (PSF) versus growth-friendly surgery and definitive fusion (GFDF). In a retrospective cohort study, children with neuromuscular EOS, age 8-11 years at index surgery with PSF or GF devices, with minimum 2-year follow-up after final fusion were identified from a multicenter database. 16 PSF and 43 GFDF patients were analyzed. Demographics were similar except PSF patients were older at index surgery and had shorter follow-up. PSF patients had greater percentage major curve correction (62% vs 38%, p = 0.001) and smaller major curve at final follow-up (23° vs 40°, p = 0.005). The GFDF group underwent over five times more surgeries (8.7 vs 1.6, p = 0.0001). Four PSF patients (25%) experienced ten complications, resulting in five unplanned returns to the operating room (UPROR) in three patients (19%). 36 GFDF patients (84%) experienced 83 complications, resulting in 45 UPRORs in 24 patients (56%). Poisson regression adjusted for age showed that the GFDF group had more complications (p = 0.001) and UPRORs (p = 0.01). Although the GFDF patients had smaller preoperative T1-T12 and T1-S1 lengths, these were similar to the PSF patients at final follow-up, indicating that the GFDF patients had greater spinal growth. PSF patients had better postoperative EOSQ-24 Financial Impact and Family Burden scores. While there was a difference in age at index surgery, PSF may be more effective than GFDF at controlling neuromuscular EOS. GFDF patients achieved more spinal growth but eight times more complications and nine times more UPRORs.


Role and mechanism of miR-4778-3p and its targets NR2C2 and Med19 in cervical cancer radioresistance.
The aim of this study was to investigate the effect of miR-4778-3p on the radiosensitivity of cervical cancer cells and to elucidate the underlying mechanism. Tissue samples were collected from eight patients with cervical cancer prior to chemoradiotherapy. MicroRNA chip analyses, RT-PCR, gene transfection, CCK8, wound healing and Transwell assays, colony-forming assay, western blot, and the Dual-Luciferase Reporter Assay System were used to evaluate the role of miR-4778-3p in cervical cancer radiosensitivity and its relationships with target molecules NR2C2 and Med19. Thirty-two differentially expressed miRNA molecules (fold-change > 2; p < 0.05) associated with cervical cancer radioresistance were identified. The expression of miR-4778-3p was significantly lower in recurrent or metastatic patients than in control subjects. In vitro studies using radioresistant HeLa and SiHa cervical cancer cell lines showed that miR-4778-3p upregulation significantly inhibited cell proliferation, invasiveness, and migration after irradiation. There was also a significant increase in apoptosis and a significant decrease in the proportion of cells at the G2/M phase. Further, miR-4778-3p upregulation led to increased expression of apoptosis-related molecules, such as Bax, Caspase-3, Caspase-8, and Caspase-9. Reporter gene assays showed that miR-4778-3p bound specifically to NR2C2 and Med19 and negatively regulated their expression. Thus, miR-4778-3p reduces the vitality, proliferation, and migration of radioresistant cervical cancer cells and may regulate the radiosensitivity of cervical cancer by targeting and regulating NR2C2 and Med19 expression.


Cryptococcus neoformans CAP59 (or Cap59p) is involved in the extracellular trafficking of capsular glucuronoxylomannan.
Several genes are essential for Cryptococcus neoformans capsule synthesis, but their functions are unknown. We examined the localization of glucuronoxylomannan (GXM) in strain B-3501 and in cap59 mutants B-4131 and C536. Wild-type strain B-3501 showed a visible capsule by India ink staining and immunofluorescence with anticapsular monoclonal antibodies (MAbs) 12A1 and 18B7. B-4131, a mutant containing a missense mutation in CAP59, showed no capsule by India ink staining but revealed the presence of capsular polysaccharide on the cell surface by immunofluorescence. The cap59 gene deletion mutant (C536), however, did not show a capsule by either India ink staining or immunofluorescence. Analysis of cell lysates for GXM by enzyme-linked immunosorbent assay revealed GXM in C536 samples. Furthermore, the epitopes recognized by MAbs 12A1, 2D10, 13F1, and 18B7 were each detected in the cytoplasm of all strains by immunogold electron microscopy, although there were differences in location consistent with differences in epitope synthesis and/or transport. In addition, the cells of B-3501 and B-4131, but not those of the cap59 deletant, assimilated raffinose or urea. Hence, the missense mutation of CAP59 in B-4131 partially hampered the trafficking of GXM but allowed the secretion of enzymes involved in hydrolysis of raffinose or urea. Furthermore, the cell diameter and volume for strain C536 are higher than those for strain B-3501 or B-4131 and may suggest the accumulation of cellular material in the cytoplasm. Our results suggest that CAP59 is involved in capsule synthesis by participating in the process of GXM (polysaccharide) export.


Chevron closing base wedge bunionectomy.
The Chevron-base wedge Association for Osteosynthesis fixated bunionectomy provides a stable, aggressive correction of the severe hallux abducto valgus deformity. It is intended for the bunion requiring a double osteotomy in order to adequately reduce both intermetatarsal and proximal articular facet angle with minimal shortening and elevation. This article presents the rationale for the procedure, technique, and a 4-year follow-up of six patients with eight Chevron-base wedge bunionectomies.


Mild deoxygenation of aromatic ketones and aldehydes over Pd/C using polymethylhydrosiloxane as the reducing agent.
Herein, a practical and mild method for the deoxygenation of a wide range of benzylic aldehydes and ketones is described, which utilizes heterogeneous Pd/C as the catalyst together with the green hydride source, polymethylhydrosiloxane. The developed catalytic protocol is scalable and robust, as exemplified by the deoxygenation of ethyl vanillin, which was performed on a 30 mmol scale in an open-to-air setup using only 0.085 mol % Pd/C catalyst to furnish the corresponding deoxygenated product in 93 % yield within 3 hours at room temperature. Furthermore, the Pd/C catalyst was shown to be recyclable up to 6 times without any observable decrease in efficiency and it exhibited low metal leaching under the reaction conditions.


Don't be scared: insert a mesh!
Mesh repair is now the gold standard technique of repair on incisional hernias. Infection of the mesh is a challenging complication of this type of repair. The risk of mesh infection has been shown to be greater in case of complicated hernia. We present the case of a 64 years old female who presented with an incarcerated incisional hernia with bowel infarction. Treated with a non absorbable mesh repair, she developed mesh infection. The infection was successively treated with simple drainage. This case and review of relevant literature seem to be an indication that mesh repair could still be considered in cases of complicated hernia. Simple drainage usually helps manage the cases of mesh infection.


Evaluation of a CD5-specific immunotoxin for treatment of acute graft-versus-host disease after allogeneic marrow transplantation.
Acute graft-versus-host disease (GVHD) is most often treated with high dose glucocorticoids, but less than half of patients have durable overall improvement. Previous phase I and phase II studies suggested that treatment with a CD5-specific immunotoxin (XomaZyme-CD5 Plus) could ameliorate symptoms of GVHD. In a randomized, double-blind trial, we compared XomaZyme-CD5 Plus and glucocorticoids versus placebo and glucocorticoids as initial therapy for 243 patients who developed acute GVHD after allogeneic marrow transplantation. The study drug (XomaZyme. CD5-Plus or an identical appearing placebo) was administered at a dose of 0.1 mg/kg body weight on each of 14 consecutive days. All patients were treated concomitantly with a standard regimen of methylprednisolone. At the time of entry on study, 94% of patients had a rash, 56% had hyperbilirubinemia, 61% had diarrhea, and 84% had nausea and vomiting. At 3, 4, and 5 weeks after starting treatment, symptom severity was less in the CD5 group than in the placebo group. At 4 weeks, 40% of patients assigned to the CD5 group had complete response compared with 25% of those assigned to the control group (P = .019). At 6 weeks, 44% of patients assigned to the CD5 group had complete response as compared with 38% in the placebo group (P = .36). Clinical extensive chronic GVHD developed in 65% of patients in the CD5 group compared with 72% in the control group (P = .35). Survival at 1 year after treatment was 49% in the CD5 group and 45% in the control group (P = .68). Side effects required close monitoring and appropriate adjustment of treatment. The combined administration of a CD5-specific immunotoxin and glucocorticoids controls GVHD manifestations more effectively than treatment with glucocorticoids alone during the first 5 weeks after starting treatment. Use of this immunotoxin does not result in any long-term clinical benefit for patients with acute GVHD.


Identification of diabetes self-management profiles in adults: A cluster analysis using selected self-reported outcomes.
The present study describes adult diabetes self-management (DSM) profiles using self-reported outcomes associated with the engagement in diabetes care activities and psychological adjustment to the disease. We used self-reported data from a community-based cohort of adults with diabetes (N = 316) and conducted a cluster analysis of selected self-reported DSM outcomes (i.e., DSM behaviors, self-efficacy and perceived empowerment, diabetes distress and quality of life). We tested whether clusters differed according to sociodemographic, clinical, and care delivery processes variables. Cluster analysis revealed four distinct DSM profiles that combined high/low levels of engagement in diabetes care activities and good/poor psychological adjustment to the disease. The profiles were differently associated with the variables of perceived financial insecurity, taking insulin treatment, having depression, and the congruence of the care received with the Chronic Care Model. The results could help health professionals gain a better understanding of the different realities facing people living with diabetes, identify patients at risk of poor outcomes related to their DSM, and lead to the development of profile-specific DSM interventions.


Structured light using carbon nanostructures driven by Kerr nonlinearities and a magnetic field.
A substantial influence of a magnetic field on the third-order nonlinear optical properties exhibited by aggregated networks of aligned carbon nanotubes (CNT) is reported by systematic measurements. A two-wave mixing was employed to explore and modulate the refractive index in the nanostructures in the nanosecond and picosecond regime. The presence of a magnetic field was able to modify the optical transmittance in the sample and the potentiality to generate structured light was proposed. Numerical simulations were conducted to analyze the magnetic field phenomena and the oscillations of the electric field in the studied sample. We discussed theoretical concepts, experimental methods, and computational tools employed to evaluate the third-order nonlinear optical properties of CNT in film form. Immediate applications of the system to modulate structured light can be contemplated.


ACP-BC: A Model for Accurate Identification of Anticancer Peptides Based on Fusion Features of Bidirectional Long Short-Term Memory and Chemically Derived Information.
Anticancer peptides (ACPs) have been proven to possess potent anticancer activities. Although computational methods have emerged for rapid ACPs identification, their accuracy still needs improvement. In this study, we propose a model called ACP-BC, a three-channel end-to-end model that utilizes various combinations of data augmentation techniques. In the first channel, features are extracted from the raw sequence using a bidirectional long short-term memory network. In the second channel, the entire sequence is converted into a chemical molecular formula, which is further simplified using Simplified Molecular Input Line Entry System notation to obtain deep abstract features through a bidirectional encoder representation transformer (BERT). In the third channel, we manually selected four effective features according to dipeptide composition, binary profile feature, k-mer sparse matrix, and pseudo amino acid composition. Notably, the application of chemical BERT in predicting ACPs is novel and successfully integrated into our model. To validate the performance of our model, we selected two benchmark datasets, ACPs740 and ACPs240. ACP-BC achieved prediction accuracy with 87% and 90% on these two datasets, respectively, representing improvements of 1.3% and 7% compared to existing state-of-the-art methods on these datasets. Therefore, systematic comparative experiments have shown that the ACP-BC can effectively identify anticancer peptides.


Interplay of Affinity and Surface Tethering in Protein Recognition.
Surface-tethered ligand-receptor complexes are key components in biological signaling and adhesion. They also find increasing utility in single-molecule assays and biotechnological applications. Here, we study the real-time binding kinetics between various surface-immobilized peptide ligands and their unrestrained receptors. A long peptide tether increases the association of ligand-receptor complexes, experimentally proving the fly casting mechanism where the disorder accelerates protein recognition. On the other hand, a short peptide tether enhances the complex dissociation. Notably, the rate constants measured for the same receptor, but under different spatial constraints, are strongly correlated to one another. Furthermore, this correlation can be used to predict how surface tethering on a ligand-receptor complex alters its binding kinetics. Our results have immediate implications in the broad areas of biomolecular recognition, intrinsically disordered proteins, and biosensor technology.


[Correlation of MDR1 and ABCG2 genetic polymorphisms with the efficacy and adverse events of irinotecan chemotherapy in patients with colorectal cancer].
To investigate the correlation of MDR1 and ABCG2 genetic polymorphisms with the efficacy and adverse events of irinotecan chemotherapy in patients with colorectal cancer (CRC). Clinical data of CRC patients treated with irinotecan-based chemotherapy in the Peking University Cancer Hospital between January 1996 and December 2011 were collected, and their blood samples were collected accordingly. Genomic DNA was extracted from blood samples. The following SNP detection of MDR1 and ABCG2 genes was conducted by direct sequencing method. The correlation of genetic SNPs with efficacy and toxicity of irinotecan treatment was further analyzed. Allele frequencies of MDR1 2677 G>T/A, ABCG2 421 C>A, 34 G>A, 376 C>T were comparable with previous studies. Genetic SNPs results from peripheral blood samples and tumor tissues were highly consistent. Patients carrying MDR1 2677 wild type had higher clinical benefit than those carrying mutant genotype, while the differences were not significant. The progression-free survival (PFS) was longer in wild-type patients as compared to mutant-type patients in second-line chemotherapy (P=0.012). There were no significant correlations between ABCG2 421 C>A, 34 G>A, 376 C>T and chemotherapy efficacy. No significant correlations were observed between MDR1 2677 G>T/A, ABCG2 421 C>A, ABCG2 34 G>A, ABCG2 376 C>T and irinotecan-related grade 3 and 4 neutropenia or diarrhea. MDR1 2677 G>T/A may be served as a biomarker in predicting the efficacy of irinotecan chemotherapy in patients with colorectal cancer.


Efficient double fragmentation ChIP-seq provides nucleotide resolution protein-DNA binding profiles.
Immunoprecipitated crosslinked protein-DNA fragments typically range in size from several hundred to several thousand base pairs, with a significant part of chromatin being much longer than the optimal length for next-generation sequencing (NGS) procedures. Because these larger fragments may be non-random and represent relevant biology that may otherwise be missed, but also because they represent a significant fraction of the immunoprecipitated material, we designed a double-fragmentation ChIP-seq procedure. After conventional crosslinking and immunoprecipitation, chromatin is de-crosslinked and sheared a second time to concentrate fragments in the optimal size range for NGS. Besides the benefits of increased chromatin yields, the procedure also eliminates a laborious size-selection step. We show that the double-fragmentation ChIP-seq approach allows for the generation of biologically relevant genome-wide protein-DNA binding profiles from sub-nanogram amounts of TCF7L2/TCF4, TBP and H3K4me3 immunoprecipitated material. Although optimized for the AB/SOLiD platform, the same approach may be applied to other platforms.


[Psychometric data of the abbreviated version of IMAGENquestionnaire; evaluation of body dissatisfaction].
Body dissatisfaction seems to be associated with eating disorders, obesity, decreasing levels of physical activity and fruit and vegetable consumption. In this study the psychometric properties of a questionnaire to measure body dissatisfaction, the IMAGEN questionnaire, have been obtained. The samples have been composed of 538 women of ages between 11 and 22 years, 494 out from different schools and 44 out diagnosed of eating disorder from hospital. The confirmatory factor analysis reduces the items from 38 in the original version to 25. The model shows a penta-factor structure: a) emotional, with five items; b) fear to gain weight, with four; c) approach to change, with three; d) perceptive, with six; and e) behavioral, with four items. Besides, it has been obtained a second order factor (cognitive-emotional) that includes the a), b) and c) previous factors. The psychometric properties such as internal consistency (alpha from 0.84 to 0.92), discriminative validity (significant difference by group) and convergent validity (significant statistical relation between IMAGE and EDI-2 factors) have been evaluated and some acceptable results have been obtained. In addition to that, the cut-offs for each questionnaire scale were established, implying that high scores in body dissatisfaction are associated with having an eating disorder. The most accurate estimates correspond to the behavioral factor of dissatisfaction (odds ratio 33.2; IC 95% = 15.1-68.3). From these results it is concluded that the reduced IMAGE questionnaire presents good psychometric properties. However, more research is needed with different samples before generalizing its use.


Note: Characterization of electrode materials for dielectric spectroscopy.
When measuring the dielectric properties of aqueous samples, the impedance of the electrode/sample interface can limit low frequency measurements. The electrode polarization problem can be reduced by increasing the effective surface area of the electrodes. In this work, impedance spectroscopy was used to characterize and compare three different electrode surfaces that can be used to mitigate this effect: platinum black, iridium oxide, and [polypyrrole/poly(styrenesulphonate)] (PPy/PSS) conducting polymer. All three materials were directly compared with a bright platinum electrode. Equivalent circuit models were used to extract the increase in the effective surface area of the electrodes: platinum black, iridium oxide and PPy/PSS increase the effective capacitance of the electrode by factors of approximately 240, 75, and 790, respectively. The practical aspects of all electrode materials are discussed. These results suggest that iridium oxide and PPy/PSS are good alternatives to the commonly used platinum black, which is prone to mechanical damage (scratches) and is potentially toxic to cells.


[Proteolytic degradation of native hemoglobin and its constituent parts--isolated subunits and globin. I. Kinetic data and the character of the process of the breakdown of native forms].
The rates of proteolytic breakdown for native human hemoglobin (Hb) in CNmet-and oxy-forms, for isolated native alpha- and beta-chains of human Hb with deprotected SH-groups and for apo-Hb--globin at constant temperature 20 degrees as well as for metHb and globin in the temperature range 4-25 degrees were studied. The proteolysis of oxy-forms of proteins was performed in the presence of CN- to prevent the appearance in solution of quickly splitted aqua and hydroxy met-forms. Pepsin (at pH 5.5), trypsin (at pH 7.0 and 8.5) and protease VI (pronase) (at pH 7.0 and 8.5) were used as proteases. The rate of proteolysis was registered simultaneously by proteolysate precipitation in concentrated salt solutions (to determine the content of the native form), by precipitation in aqueous solution of trichloroacetic or perchloric acid and by colouring the terminal NH2-groups by ninhidrin in the total proteolysate. For most cases the data of all the three independent methods fell on a single kinetic curve, each pair protein--protease being represented by their individual curves. Therefore the breakdown of all the protein studied possesses a burst-like ("one-by-one", "all-or-none") character. The protein resistance to the attack by proteolytic enzymes increases in the following order: globin less than oxy-alpha-chain less than metHb less than oxy-beta-chain less than HbO2 congruent to CNmetHb. The use of control repeated proteolysis has made it possible to prove that differences in the rate of proteolytic degradation are not the consequence of spontaneous denaturation of the least unstable forms of proteins in the course of proteolytic reaction but are predetermined by the conformational state of the native macromolecule.


A COLQ missense mutation in Labrador Retrievers having congenital myasthenic syndrome.
Congenital myasthenic syndromes (CMSs) are heterogeneous neuromuscular disorders characterized by skeletal muscle weakness caused by disruption of signal transmission across the neuromuscular junction (NMJ). CMSs are rarely encountered in veterinary medicine, and causative mutations have only been identified in Old Danish Pointing Dogs and Brahman cattle to date. Herein, we characterize a novel CMS in 2 Labrador Retriever littermates with an early onset of marked generalized muscle weakness. Because the sire and dam share 2 recent common ancestors, CMS is likely the result of recessive alleles inherited identical by descent (IBD). Genome-wide SNP profiles generated from the Illumina HD array for 9 nuclear family members were used to determine genomic inheritance patterns in chromosomal regions encompassing 18 functional candidate genes. SNP haplotypes spanning 3 genes were consistent with autosomal recessive transmission, and microsatellite data showed that only the segment encompassing COLQ was inherited IBD. COLQ encodes the collagenous tail of acetylcholinesterase, the enzyme responsible for termination of signal transduction in the NMJ. Sequences from COLQ revealed a variant in exon 14 (c.1010T>C) that results in the substitution of a conserved amino acid (I337T) within the C-terminal domain. Both affected puppies were homozygous for this variant, and 16 relatives were heterozygous, while 288 unrelated Labrador Retrievers and 112 dogs of other breeds were wild-type. A recent study in which 2 human CMS patients were found to be homozygous for an identical COLQ mutation (c.1010T>C; I337T) provides further evidence that this mutation is pathogenic. This report describes the first COLQ mutation in canine CMS and demonstrates the utility of SNP profiles from nuclear family members for the identification of private mutations.


Predicting the risk of high-grade bladder cancer using noninvasive data.
To examine the hypothesis that the risk of high-grade bladder cancer can be predicted using noninvasively obtained data. We retrospectively analyzed the database of 431 patients that had transurethral resection of first-time bladder tumors between June 1998 and December 2009. Pre-operative parameters evaluated were: patients' age; gender; sonographic tumor diameter, number and location of tumor inside the bladder; presence of hydronephrosis, and results of urinary cytology. Parameters that showed significance in multivariate analysis were incorporated into the nomogram. Multivariate analysis of the data showed that patient's age, the presence of hydronephrosis, sonographic tumor diameter (risk of a high-grade tumor: 14, 29, 43.3, 55.7 and 69.4% at diameters: 0.5-1.5, 1.6-2, 2.1-2.5, 2.6-3 and >3 cm, respectively), location of tumor in the bladder (risk of high-grade tumor: 28.8, 47, 67.5 and 90.5% in the lateral walls, posterior/base, anterior and dome, respectively), and urinary cytology were all highly significant and independent predictors of high-grade tumors. A nomogram constructed using these variables scored an area of 0.853 in the ROC curve. The risk of high-grade bladder tumor can be accurately predicted using non-invasively obtained information. This prediction can help to triage patients with newly detected bladder cancer for biopsy.


Ultrasound reverses chemoresistance in breast cancer stem cell like cells by altering ABCG2 expression.
Doxorubicin (DOX) resistance in breast cancer largely results from the breast cancer stem cell like cells (BCSCs) which could be targetted to improve the efficacy of chemotherapy. Cell permeabilization using microbubbles (MBs) and ultrasound (US) have the potential for delivering molecules into the cytoplasm. We aim to evaluate a new methodology of US on BCSCs. First, our findings indicated that ALDHA1+ spheres which were derived from fresh primary breast cancer samples displayed stem cell like features and were resistant to DOX. In patient cohort, we revealed the presence of a variable fraction of ALDHA1+cells in nine out of ten. We, for the first time, showed a new US-MB treatment condition which could be used on ALDHA1+ BCSCs by fluorescence measurement and calcein assay. Next, we demonstrated the efficacy of combined treatment on human BCSCs in vitro and in vivo using DOX and US-MB: the combined treatment with much reduced drug dosage significantly suppressed the stem cell like features of BCSCs and induced BCSCs apoptosis. Furthermore, we suggested that decreased ABCG2 level might be one of the mechanisms by which US-MB medicated DOX treatment. In conclusion, this new US-MB treatment condition has clinical potential in breast cancer therapy by targetting BCSCs; thereby holding benefits for breast cancer patients.


Evidence and Evidence-Based Practices: Are We There Yet?
The purpose of this article is to move the field of intellectual and closely related developmental disabilities (IDD) towards a better understanding of evidence and evidence-based practices. To that end, we discuss (a) different perspectives on and levels of evidence, (b) commonly used evidence-gathering strategies, (c) standards to evaluate evidence, (d) the distinction between internal and external validity, and (e) guidelines for establishing evidence-based practices. We also describe how the conceptualization and use of evidence and evidence-based practices are changing to accommodate recent trends in the field.


Response of nitrogen metabolism in preparturient dairy cows to methionine supplementation.
Three multiparous Holstein cows (607 kg of BW) were surgically prepared with an elevated carotid artery and indwelling catheters in the hepatic, portal, and two mesenteric veins to study the effects of methionine supplementation on amino acid metabolism during the last 2 wk of pregnancy. The study began 15 d before the expected calving date. Dietary treatments were Control (1.53 Mcal NE(l)/kg, 15.6% CP, and 40% ruminally undegradable protein) and Control supplemented with 60 g/d of ruminally protected methionine (MET, supplying 39 g/d of DL-methionine and approximately 18 g/d of methionine available for intestinal absorption). Each cow received both dietary treatments in a crossover design. Cows were fed once daily. After 5 d on treatment, a blood flow marker (para-aminohippurate) was infused into a mesenteric vein, and arterial, portal, and hepatic blood samples were obtained at 0, 2, 6, 12, and 18 h after feeding. Net flux of methionine was calculated as the plasma arteriovenous difference multiplied by plasma flow. Dry matter intake (10.8 kg/d) and portal (824 L/h) and hepatic (995 L/h) plasma flows were not affected (P > .10) by treatment. Arterial plasma concentration of methionine was greater (P = .10) with MET (27.67 microM) than with Control (16.42 microM). Net portal absorption of methionine increased (P = .10) with MET (26.2 g/d) compared with Control (9.5 g/d). The net portal methionine flux was negatively correlated (r = -.59; P < .001) with arterial urea concentrations. Net flux of methionine across splanchnic tissues shifted (P = .06) from a net uptake with Control (4 g/d) to a net output with MET (11 g/d). Therefore, MET increased by 15 g/d the methionine supply to the rest of the body. The net uptake of methionine by splanchnic tissues observed with Control indicated a net mobilization of methionine by peripheral tissues. Results indicate that methionine was the limiting amino acid with Control and that MET was beneficial because it increased methionine supply to peripheral tissues and reduced arterial urea concentrations.


Primary Colonic Epithelioid Angiosarcoma with Hepatic Metastasis: A Case Report.
Colonic angiosarcoma is an extremely rare and aggressive malignant tumor with poor prognosis. We report a case of colonic epithelioid angiosarcoma with colonic obstruction and rapidly progressive hepatic metastasis in a 44-year-old female. Abdominal CT revealed a heterogeneously enhancing irregular mass in the ascending colon, causing proximal bowel distension. The patient underwent surgery, and histopathological examination revealed a poorly differentiated carcinoma. A follow-up liver dynamic MRI after 4 months revealed newly developed diffusely scattered numerous small nodules in both hepatic lobes with peripheral and nodular marked arterial hyperenhancement, raising the suspicion of hepatic angiosarcoma. A pathologic second opinion was obtained, and additional immunohistochemistry revealed colonic epithelioid angiosarcoma. The patient showed progressive hepatic metastasis on follow-up abdominal CT after 6 months and died 8 months after initial diagnosis. We describe an educational case of colonic angiosarcoma, a rare malignant tumor, with rapidly progressive hepatic metastasis that showed radiologic findings suggestive of angiosarcoma and enabled a re-diagnosis for proper treatment and prognosis prediction.


Nutritional Alert in hospitalized patients.
The objective of the study is to assess a nutritional risk alert using an Informatics System in hospitalized patients versus the conventional methodology. We studied 400 medical patients at the Belgrano Hospital Critical Care Unit. We considered two groups of 200 patients. In the first the group (Control Group), nutritional risk was diagnosed in a traditional way. In the second group (Alarm Group), nutritional risk was diagnosed with an alert system. The alert was triggered when a patient showed low levels of, at least, two of the following variables: albumin, cholesterol, triglyceride, lymphocyte or low body mass index (BMI). Nutritional risk was detected in 20.3% of the Control Group patients while, in the Alert Group, nutritional risk was detected in 34.3% of the patients; the difference between the two groups was significant (p<0.001), with a sensitivity rate of 99% and a specificity rate of 98%. Malnutrition is more easily detected when using an alert system.


Highlights in pathogenic fungal biofilms.
A wide variety of fungi have demonstrated the ability to colonize surfaces and form biofilms. Most studies on fungal biofilms have focused on Candida albicans and more recently, several authors have reported the involvement of other genera of yeasts and Candida species, as well as of filamentous fungi in the formation of biofilms, including: Cryptococcus neoformans, Cryptococcus gattii, Rhodotorula species, Aspergillus fumigatus, Malassezia pachydermatis, Histoplasma capsulatum, Paracoccidioides brasiliensis, Pneumocystis species, Coccidioides immitis, Fusarium species, Saccharomyces cerevisiae, Trichosporon asahii, Mucorales and Blastoschizomyces. There is a current interest in describing the particular characteristics of the biofilm formation by of these fungi. A major concern is the control of biofilms, requiring knowledge of the biofilm mechanisms. However, our knowledge of these microbial communities is limited, due to the complexity of these systems and metabolic interactions that remain unknown. This mini-review aims to highlight recently discovered fungal biofilms and to compare them with the current knowledge on biofilms. This manuscript is part of the series of works presented at the "V International Workshop: Molecular genetic approaches to the study of human pathogenic fungi" (Oaxaca, Mexico, 2012).


[Thrombogenic factors in the elderly. Evaluation of the plasma concentrations of thromboxane B2 and antithrombin III].
The aim of this study is to prove the existence of a major tendency of platelet aggregation in elderly patients compared to medium-aged adults and, also, to detect whether it is affected by the presence of diabetes mellitus. Plasmatic concentrations of B2 thromboxane (TXB2) and antithrombin III (AT III) were determined in 73 elderly patients of both sexes; 56 without metabolic disease known (Group a) and 17 diabetic patients, 7 type I (Group bI) and 10 of type II (Group bII); and 12 healthy adults (control group). Medium plasmatic concentration of TXB2 was significantly higher (p less than 0.001) in Group a (55 +/- 14 ng/ml) compared to the control group (37 +/- 9 ng/ml) and there was no difference between Group bI (53 +/- 19 ng/ml) and bII (57 +/- 15 ng/ml). No variations were noted in ATIII concentration between the adults (27.4 +/- 2.3 mg/dl) and elderly patients (a = 29.6 +/- 4.4, bI = 29 +/- 2.6, bII = 31.2 +/- 5.9 mg/dl). In elderly patients, there appears to be a state of platelet pro-aggregation without influence of any risk factor, such as diabetes. This could explain the thrombogenic tendency of this age group.


Limosilactobacillus caccae sp. nov., a new bacterial species isolated from the human gut microbiota.
Strain Marseille-P3519T isolated from the fecal flora of a 25-year-old healthy French woman was a Gram-positive anaerobic bacterium, non-motile and non-spore forming. The 16S rRNA gene sequence of Marseille-P3519 showed 97.73% of sequence similarity with Limosilactobacillus reuteri DSM 20016, the closest species, phylogenetically. Furthermore, the average nucleotide identity of strain Marseille-3519 with its closest related species was 75.8% that was very below the recommended threshold (>95-96%). Its genome had 2 237 367 bp with 45.42 mol% of G + C content. Major fatty acids were C16:0 (50.8%), C18:1n9 (18.0%), C18:2n6 (9.8%) and C19:1n9 (8.9%). It was catalase negative and fermented glycerol, glucose, fructose, D-maltose, lactose and mannose. These findings support that strain Marseille-P3519 ( = CSURP3519 = CECT 30110) is a new member of the genus Limosilactobacillus for which the name Limosilactobacillus caccae sp. nov., is proposed.


AATYK1A phosphorylation by Cdk5 regulates the recycling endosome pathway.
Trafficking of recycling endosomes (REs) is regulated by the small GTPase, Rab11A; however, the regulatory mechanism remains elusive. Apoptosis-associated tyrosine kinase 1A (AATYK1A) is a Ser/Thr kinase expressed highly in brain. We have recently shown that AATYK1A localizes to Rab11A-positive RE and is phosphorylated at Ser34 by cyclin-dependent kinase 5 (Cdk5). Here, we have investigated a role of AATYK1A and its phosphorylation in recycling endosomal trafficking using Chinese hamster ovary-K1 (CHO-K1) cells. AATYK1A localizes predominantly to Rab11A-positive pericentrosomal endocytic recycling compartment (ERC). Phosphorylation at Ser34 of AATYK1A disrupts its accumulation in the pericentrosomal ERC. Consistently, phosphorylation-mimic mutant (AATYK1A-S34D) did not accumulate in the ERC and additionally attenuated ERC formation. ERC formation suppression can be reversed by constitutively active Rab11A-Q70L, suggesting a functional link between AATYK1A phosphorylation and Rab11A activity. Although no direct interaction between AATYK1A and Rab11A could be detected, the exchange of guanine nucleotides bound to Rab11A was significantly reduced in the presence of the phosphorylation-mimic AATYK1A-S34D. Together, our results reveal a regulatory role for AATYK1A in the formation of pericentrosomal ERC. They furthermore indicate that Cdk5 can disrupt ERC formation via Ser34 phosphorylation of AATYK1A. Finally, our data suggest a mechanism by which AATYK1A signaling couples Cdk5 to Rab11A activity.


Oocyte-targeted deletion reveals that hsp90b1 is needed for the completion of first mitosis in mouse zygotes.
Hsp90b1 is an endoplasmic reticulum (ER) chaperone (also named Grp94, ERp99, gp96,Targ2, Tra-1, Tra1, Hspc4) (MGI:98817) contributing with Hspa5 (also named Grp78, BIP) (MGI:95835) to protein folding in ER compartment. Besides its high protein expression in mouse oocytes, little is known about Hsp90b1 during the transition from oocyte-to-embryo. Because the constitutive knockout of Hsp90b1 is responsible for peri-implantation embryonic lethality, it was not yet known whether Hsp90b1 is a functionally important maternal factor. To circumvent embryonic lethality, we established an oocyte-specific conditional knockout line taking advantage of the more recently created floxed Hsp90b1 line (Hsp90b1(flox), MGI:3700023) in combination with the transgenic mouse line expressing the cre recombinase under the control of zona pellucida 3 (ZP3) promoter (Zp3-cre, MGI:2176187). Altered expression of Hsp90b1 in growing oocytes provoked a limited, albeit significant reduction of the zona pellucida thickness but no obvious anomalies in follicular growth, meiotic maturation or fertilization. Interestingly, mutant zygotes obtained from oocytes lacking Hsp90b1 were unable to reach the 2-cell stage. They exhibited either a G2/M block or, more frequently an abnormal mitotic spindle leading to developmental arrest. Despite the fact that Hspa5 displayed a similar profile of expression as Hsp90b1, we found that HSPA5 and HSP90B1 did not fully colocalize in zygotes suggesting distinct function for the two chaperones. Consequently, even if HSPA5 was overexpressed in Hsp90b1 mutant embryos, it did not compensate for HSP90B1 deficiency. Finally, further characterization of ER compartment and cytoskeleton revealed a defective organization of the cytoplasmic region surrounding the mutant zygotic spindle. Our findings demonstrate that the maternal contribution of Hsp90b1 is critical for the development of murine zygotes. All together our data indicate that Hsp90b1 is involved in unique and specific aspects of the first mitosis, which brings together the maternal and paternal genomes on a single spindle.


Antibacterial activities of blueberry and muscadine phenolic extracts.
Phenolics are one category of phyto-antimicrobials that refer to the antimicrobial substances extracted from plant sources. This study was undertaken to determine the influence of blueberry and muscadine phenolic extracts on the growths of 2 important foodborne bacterial pathogens, Salmonella Enteritidis and Listeria monocytogenes. Cells of S. Enteritidis (n = 4) or L. monocytogenes (n = 4) strains were inoculated (3 log CFU/mL) into tryptic soy broth (TSB) supplemented with 46.25 ppm of muscadine phenolics and 24 ppm of blueberry phenolics, respectively. The inoculated and un-inoculated broth with or without the supplemented phenolics were incubated at 37 °C for 24 h. Samples were drawn periodically, and cell populations of Salmonella and Listeria were determined on tryptic soy agar (TSA). It was observed that Salmonella was relatively more susceptible than Listeria to the phenolic extracts used in the study. The growth of Salmonella was significantly inhibited in all samples at all sampling points except for the sample that was supplemented with muscadine water extract and drawn at the 24-h sampling point. Blueberry phenolics were relatively more effective than muscadine phenolic extracts in inhibiting the growth of Salmonella. One tested strain of Listeria was more susceptible to ethanol than water phenolic extracts. The study revealed the potentials and limitations of using blueberry and muscadine phenolics to control the growths of selected Salmonella and Listeria strains.


Comparative testing of monoclonal antibodies against Plasmodium falciparum sporozoites for ELISA development.
Ten monoclonal antibodies developed against Plasmodium falciparum sporozoites at four institutions were evaluated for use in an enzyme-linked immunosorbent assay (ELISA). Four of the antibodies were eliminated because of their low sensitivity or requirement for high concentrations of capture antibody, while an additional four were rejected because they exhibited cross-reactivity with P. berghei sporozoites. Of the two remaining monoclonal antibodies, that designated 2A10 had the highest sensitivity, a requirement for lower concentrations of capture antibody, and had been tested successfully against sporozoites from a wider range of geographical areas than the others. Use of this monoclonal antibody in a standardized ELISA method gave a test ten times more sensitive than previously reported for P. falciparum sporozoites and its detection limit was less than 100 sporozoites per mosquito.


1971 - 2011: Forty year history of scope expansion into medical eye care.
The focus of this paper is to provide a historical timeline for many of the well over 180 incremental scope of practice expansion and amplification legislative successes achieved in the United States (U.S.) during the 40-year period 1971 - 2011 that cumulatively expanded optometry into medical eye care. This paper also serves to update the historical timeline of scope of practice amplification legislation enacted after the year 1999, which was so comprehensively described up to that point by Dr. Melvin D. Wolfberg.


Combination approaches to potentiate immune response after photodynamic therapy for cancer.
Photodynamic therapy (PDT) has been used as a cancer therapy for forty years but has not advanced to a mainstream cancer treatment. Although it has been shown to be an efficient way to destroy local tumors by a combination of non-toxic dyes and harmless visible light, it is its additional effects in mediating the stimulation of the host immune system that gives PDT great potential to become more widely used. Although the stimulation of tumor-specific cytotoxic T-cells that can destroy distant tumor deposits after PDT has been reported in some animal models, it remains the exception rather than the rule. This realization has prompted several investigators to test various combination approaches that could potentiate the immune recognition of tumor antigens that have been released after PDT. This review will cover these combination approaches using immunostimulants including various microbial preparations that activate Toll-like receptors and other receptors for pathogen-associated molecular patterns, cytokines growth factors, and approaches that target regulatory T-cells. We believe that by understanding the methods employed by tumors to evade immune response and neutralizing them, more precise ways of potentiating PDT-induced immunity can be devised.