Chemotaxonomic diagnostics: combining sucrose-water agar with TLC to discriminate Fusarium graminearum 3-acetyl-DON and 15-acetyl-DON chemotypes.
Twelve randomly-selected isolates of Fusarium graminearum that produce 3-acetyl-deoxynivalenol (3-ADON) or 15-acetyl-deoxynivalenol (15-ADON) were screened by thin-layer chromatography (TLC) for their ability to produce ADON and zearalenone (ZEA) mycotoxins when grown on water agar containing different concentrations of sucrose. The results showed the ability of the F. graminearum 3-ADON chemotype population to produce DON and ZEA at a lower concentration range of sucrose (5-7%) compared with the 15-ADON chemotype (30-40%). The former distinction allows for sucrose-water agar to be employed as a rapid and simple differential medium, where two separate sucrose-gradient concentrations discriminate 3-ADON from 15-ADON populations. In the light of the shift in sugar concentrations occurring during the process of grain formation and maturation, the difference in mycotoxin production between the two populations is discussed with respect to predicting Fusarium head blight (FHB) epidemiology and accumulation of DON and ZEA.


nolMNO genes of Bradyrhizobium japonicum are co-transcribed with nodYABCSUIJ, and nolO is involved in the synthesis of the lipo-oligosaccharide nodulation signals.
A host-inducible lacZ fusion was mapped down-stream of the nodYABCSUIJ operon in Bradyrhizobium japonicum strain USDA110. Sequencing of this region identified three novel genes, nolMNO. RNA dot blot analysis showed that nolO transcription is nodD1-dependent and that a polar mutation in nodS, located 5 kilobases upstream of nolO, blocks the transcription of nolO. Coupled with the host-inducible nature of nolO expression, these results indicate that nolMNO are part of a 9-kilobase operon, nodYABCSUIJnolMNO. The lipo-oligosaccharide nodulation signals produced by strains SL67 (nolO-) and SL65(nolNO-) were purified, and their chemical structures were determined. In addition to the wild-type signal molecules, both mutants produced modified compounds that are not produced by the parent strain USDA110. The most prevalent difference observed was the absence of the 2-O-methylfucosyl residue from the mutant structures. In addition, metabolites were found in which the N-acetylglucosamine residue at the reducing end was glycosidically linked to glycerol. These alterations in the profiles of nodulation signals produced by strains SL67 and SL65 were accompanied by reduced nodulation efficiency on all hosts tested.


Cycling fatalities in Western Australia.
To present a review of the circumstances of 64 cycling fatalities in Western Australia (WA) during the nine-year period 1984-1992, and the major injuries sustained by the cyclists. The number of deaths was ascertained from lists maintained by the WA Police Department, Forensic Pathology Division of the Health Department and the Coroner's office. Individual case files held by the Coroner's office and Forensic Pathology Division were then examined for details of the circumstances of each incident, epidemiological data, and medical and toxicological findings. Most of the 64 fatalities (59%) occurred in children or young adults less than 20 years of age. Most fatal injuries (54%) were to the head alone. Forty-seven of the deaths involved collision with a motor car. Alcohol was demonstrated in the blood of five of the 30 cases in which analysis of blood alcohol level was performed. On average, 7.8 cyclists died annually in WA before the introduction of legislation for the compulsory wearing of safety helmets while cycling. In the 12-month period following this legislation, there was only one cycling death. Most cycling deaths occur in children, primarily from head injury resulting from impact with a motor car. The enactment of legislation for the compulsory wearing of cycling helmets has been associated with a fall in cyclist fatalities.


Detection and frequency of recombination in tomato-infecting begomoviruses of South and Southeast Asia.
Tomato-infecting begomoviruses are widely distributed across the world and cause diseases of high economic impact on wide range of agriculturally important crops. Though recombination plays a pivotal role in diversification and evolution of these viruses, it is currently unknown whether there are differences in the number and quality of recombination events amongst different tomato-infecting begomovirus species. To examine this we sought to characterize the recombination events, estimate the frequency of recombination, and map recombination hotspots in tomato-infecting begomoviruses of South and Southeast Asia. Different methods used for recombination breakpoint analysis provided strong evidence for presence of recombination events in majority of the sequences analyzed. However, there was a clear evidence for absence or low Recombination events in viruses reported from North India. In addition, we provide evidence for non-random distribution of recombination events with the highest frequency of recombination being mapped in the portion of the N-terminal portion of Rep. The variable recombination observed in these viruses signified that all begomoviruses are not equally prone to recombination. Distribution of recombination hotspots was found to be reliant on the relatedness of the genomic region involved in the exchange. Overall the frequency of phylogenetic violations and number of recombination events decreased with increasing parental sequence diversity. These findings provide valuable new information for understanding the diversity and evolution of tomato-infecting begomoviruses in Asia.


Determination of total safranal by in situ acid hydrolysis in supercritical fluid media: Application to the quality control of commercial saffron.
A procedure allowing hydrolysis reactions to be conducted in a dynamic supercritical-CO2 medium was developed for quantifying total safranal (viz. free safranal present in the sample+safranal resulting from picrocrocin hydrolysis), which are the main component of the essential oil and responsible for the characteristic aroma of saffron. The proposed method allows total safranal amounts over the ranges 0.05-1.5 mg mL(-1) to be determined. The standard deviation achieved was 2%. This method was applied to the determination of safranal in natural saffron samples. The results obtained were compared with the "safranal value" total index, which is widely used as a quality measure of saffron products. The comparison revealed that the proposed method provides useful information not contained in the safranal value, based on the fact that, some samples with a high "safranal index" contain low concentrations of safranal. The proposed method is very useful for quality control in commercial saffron samples.


Analysis of postmortem DNA degradation by single-cell gel electrophoresis.
One of the most important longstanding problems in the field of forensic medicine is the determination of the time of death upon the discovery of a possible homicide victim. With a majority of homicide victims discovered within the first 48h, it is critically important to be able to determine time of death quickly, and with accuracy and precision. Current methods of determining postmortem interval (PMI) vary, but none can provide better than an 8-h window time estimate. In this paper, the potential application of single-cell gel electrophoresis (SCGE), also known as the comet assay, to evaluate postmortem cell death processes, specifically nuclear DNA fragmentation, is assessed. Upon the death of an organism, internal nucleases contained within the cells should cause chromosomal DNA to degrade into increasingly smaller fragments over time, and if these fragments can be isolated and visualized, the fragmentation should prove to be measurable and quantifiable. An original study providing proof of the concept of postmortem DNA fragmentation between early and late time periods was conducted using human leukocytes. With an established trend seen in the leukocyte results, this study was then expanded using a porcine animal model, over a longer time period, with more frequent time-points evaluated. DNA degradation in all samples was revealed by SCGE and quantified by the use of DNA-specific quantitative stains, as measured by digital camera affixed to a microscope. The comet 'tail-moment' gave a measure of the proportion of fragmented to non-fragmented DNA, while the 'tail-length' provided the relative size of degraded DNA fragments. In both models, an increase in DNA fragmentation was found to correlate with an increased PMI from 0 to 56h postmortem, as evaluated by comet-tail-moment and by comet-tail-length, with tail-length providing the strongest statistical correlation, based upon regression analysis. The postmortem DNA fragmentation observed in this study, reveals a sequential, time-dependent process with the potential for use as a predictor of PMI in homicide cases.


Creating Patient-Specific Neural Cells for the In Vitro Study of Brain Disorders.
As a group, we met to discuss the current challenges for creating meaningful patient-specific in vitro models to study brain disorders. Although the convergence of findings between laboratories and patient cohorts provided us confidence and optimism that hiPSC-based platforms will inform future drug discovery efforts, a number of critical technical challenges remain. This opinion piece outlines our collective views on the current state of hiPSC-based disease modeling and discusses what we see to be the critical objectives that must be addressed collectively as a field.


Editorial: Moving From Innovation to Validation With Independent Sample Testing.
It is well established that family history is the most important known risk factor for the development of a mood disorder.1 Approximately 25% of at-risk offspring will develop a mood disorder1,2; yet, this is only slightly higher than the lifetime rate of a mood disorder overall of 21.4%.3 These statistics suggest heterogeneity within mood disorders generally and within people at familial risk. More information, particularly in the form of trait-level biomarkers, is sorely needed to accurately predict mood disorder risk and development. Trait-level biomarker identification may also help to decrease the well-known delay in accurately diagnosing bipolar disorders specifically,4 which could ultimately improve treatment and outcomes. Nimarko et al.5 aimed to identify trait-level biological markers of social reward-related neural vulnerability that distinguish youths genetically at risk for mood disorder. Nimarko and colleagues reported a well-designed study consisting of extremely well-matched youths who, by virtue of parentage, were at risk of bipolar disorder, were at risk of major depressive disorder, or were healthy (control group).5 The sample sizes reflected the challenges associated with doing this type of research with children, but the importance of the relationships was clear and strong. Specifically, the results showed reduced bilateral putamen activity (left: z = 4.39, p = .003; right: z = 4.56, p < .001) and reduced left putamen connectivity with right posterior cingulate cortex (z = 4.04, p < .01) and left anterior cingulate cortex (z > 3.64, p < .001) during an emotional face social reward task as potential trait markers that distinguished at-risk samples.5 In another study, similarly reduced reward-related connectivity in genetically at-risk youths for bipolar disorder relative to at-risk and healthy groups was shown to monetary reward and loss.6 Importantly, in both of these studies, reward-related connectivity group differences were observed before threshold symptoms developed, suggesting that early identification and intervention may be possible. In addition, this reduced reward-related connectivity was not related to subthreshold symptoms in either study. Taken together, these findings suggest that social reward and monetary reward and loss may be interconnected at a neural level in youths genetically at risk for bipolar disorder before symptom development. Exploratory analyses in Nimarko et al.5 also showed that reduced reward-related connectivity may be a trait-level biomarker of risk of future conversion to an Axis I disorder over time (bipolar disorder risk: hazard ratio = 8.28, p < .01); major depressive disorder risk: hazard ratio = 2.31, p = .02). Although Acuff et al.6 did not address the longitudinal question of conversion directly, the authors speculated that reduced reward-related connectivity was related to increased risk of future bipolar disorder. This serendipitous independent replication of reward-related connectivity is a promising lead, but repeated testing in independent samples is needed. However, direct independent sample testing is uncommon in psychiatry and in psychiatric neuroscience.7,8 This is partially due to factors such as time and money, but also to the value placed on innovation, which is prized by funding agencies, journal editors, and society. Independent sample testing is not, by definition, innovative, and while innovation is an important cornerstone of science and should be encouraged, we also need to test previously identified findings using independent samples.


Investigation into the relationship between the passive flexibility and active stiffness of the ankle plantar-flexor muscles.
The purpose of the study was to investigate the relationship between measurements of passive flexibility and active stiffness of the ankle plantar-flexor muscles. The study was a correlation design. Flexibility has passive and active components. Little information is available regarding the relationship of these measurements in terms of the information that they yield on the state of the muscle-tendon unit. Free oscillation data representing active stiffness was obtained using applied masses equivalent to 30%, 60% and 90% of the subject's maximal voluntary contraction. The angle of dorsiflexion, representing passive flexibility, was measured in standing using computer digitisation to obtain the angle.Results. Mean active stiffness values were 14280 N/m (30% maximal voluntary contraction), 22260 N/m (60% maximal voluntary contraction) and 28010 N/m (90% maximal voluntary contraction). Dorsiflexion measurement gave a mean of 34.3 degrees (S.D. 4.8). Correlation's obtained for the association between range of motion and active stiffness were r=0.01 (30% maximal voluntary contraction), r=0.09 (60% maximal voluntary contraction) and r=0.04 (90% maximal voluntary contraction). Moderate reliability coefficients of 0.71 (30% maximal voluntary contraction), 0.78 (60% maximal voluntary contraction) and 0.68 (90% maximal voluntary contraction) were obtained. The results imply that measurements of passive flexibility and active stiffness of the lower leg musculature are independent measures of components of muscle-tendon unit flexibility. Flexibility is a construct with different components of measurement. Etiological studies typically relate static flexibility measurements to injury with conflicting outcomes. This study suggests that static and active flexibility measures yield different information about the muscle tendon unit of the ankle plantar flexors, and that researchers should consider this point in the design of etiological studies.


Attracting and retaining qualified nurses in aged and dementia care: outcomes from an Australian study.
To identify key issues and factors affecting retention of qualified nurses who care for older people and persons with dementia in Australian acute, subacute, community and residential health-care settings. As the number of older people with chronic conditions needing health care continues to increase research is needed to optimize nurse retention. Qualified nurses were surveyed with a set of items derived from four published nurse workforce questionnaires (Cronbach's alpha range 0.75-0.96). There were 3983 complete responses and 10 focus groups with 58 volunteer survey respondents. In addition to reporting a number of workplace issues, nurses also reported reasonable levels of satisfaction. Intrinsic factors related to caregiving, work relations and colleague support. Extrinsic factors included professional opportunities and organisational support. Altruism is a primary motivation for choosing to nurse older people and persons with dementia. Nurses are most positive when they feel valued and supported by their organisation and colleagues, through education, training, supervision, mentoring opportunities and appropriate remuneration. Nursing managers need to take positive steps to address the organisational factors outlined in this paper that either inhibit or promote nurse retention.


[Teratoma of the testicle with very highly differentiated structures].
A teratoma is described that was composed of differentiated derivatives of the germ layers--ectoderm (central and peripheral nervous systems, epiphysis, adenohypophysis, and skin with appendages), endoderm (bronchial and intestinal epithelia), and mesenchyme (fibrous and smooth-muscle tissue, cartilage, and bone) and of mesoderm (mesonephros).


Identification and characterization of plasmid-borne erm(T) macrolide resistance in group B and group A Streptococcus.
One hundred and seven group B Streptococcus (GBS) isolates and 344 group A Streptococcus (GAS) isolates were collected between 2005 and 2009 from 2 area hospitals and studied for resistance to erythromycin (ERY) and clindamycin (CLI) and the presence of the erm(T) macrolide resistance gene. The erm(T) gene was found in 5 (8%) of 61 erythromycin nonsusceptible GBS isolates and in 22 (55%) of 40 erythromycin nonsusceptible GAS isolates. The erm(T) gene in all 27 GBS/GAS erm(T) gene-positive isolates was located on a plasmid. Three erm(T) gene-positive plasmids were DNA sequenced. Two plasmids (1 each from GBS and GAS isolates) were both 4967 bp in size, contained the erm(T) gene, and differed by only 2 base pairs, suggesting interspecies horizontal transfer of the erm(T) gene containing plasmid. The third (GBS) plasmid was 6825 bp in size and contained GBSi1, a group II bacterial intron, as well as the erm(T) gene. Pulsed-field gel electrophoresis of all 27 erm(T) gene containing isolates and a selection of erm(T) gene-negative isolates indicated possible clonal expansion among erm(T) gene containing GAS isolates, but not among the 5 erm(T) gene-positive GBS isolates.


The SPID-GBA study: Sex distribution, Penetrance, Incidence, and Dementia in GBA-PD.
To provide a variant-specific estimate of incidence, penetrance, sex distribution, and association with dementia of the 4 most common Parkinson disease (PD)-associated GBA variants, we analyzed a large cohort of 4,923 Italian unrelated patients with primary degenerative parkinsonism (including 3,832 PD) enrolled in a single tertiary care center and 7,757 ethnically matched controls. The p.E326K, p.T369M, p.N370S, and p.L444P variants were screened using an allele-specific multiplexed PCR approach. All statistical procedures were performed using R or Plink v1.07. Among the 4 analyzed variants, the p.L444P confirmed to be the most strongly associated with disease risk for PD, PD dementia (PDD), and dementia with Lewy bodies (DLB) (odds ratio [OR] for PD 15.63, 95% confidence interval [CI] = 8.04-30.37, p = 4.97*10-16; OR for PDD 29.57, 95% CI = 14.07-62.13, p = 3.86*10-19; OR for DLB 102.7, 95% CI = 31.38-336.1, p = 1.91*10-14). However, an unexpectedly high risk for dementia was conferred by p.E326K (OR for PDD 4.80, 95% CI = 2.87-8.02, p = 2.12*10-9; OR for DLB 12.24, 95% CI = 4.95-30.24, p = 5.71*10-8), which, on the basis of the impact on glucocerebrosidase activity, would be expected to be mild. The 1.5-2:1 male sex bias described in sporadic PD was lost in p.T369M carriers. We also showed that PD penetrance for p.L444P could reach the 15% at age 75 years. We report a large monocentric study on GBA-PD assessing mutation-specific data on the sex distribution, penetrance, incidence, and association with dementia of the 4 most frequent deleterious variants in GBA.


Carbonylative cycloaddition between two different alkenes enabled by reactive directing groups: expedited construction of bridged polycyclic skeletons.
A novel palladium-catalyzed highly selective hydrocarbonylative cycloaddition reaction with two different alkenes in the presence of CO enabled by a reactive directing-group is developed, which offers efficient and convenient access to lactone-containing bridged polycyclic compounds in high yield with high chemo- and stereoselectivities.


Speech processor data logging helps in predicting early linguistic outcomes in implanted children.
To analyse the value of listening-data logged in the speech processor on the prediction of the early auditory and linguistic skills in children who received a cochlear implant in their first 2 years of life. Prospective observational non-randomized study. Ten children with profound congenital sensorineural hearing loss were included in the study. The mean age at CI activation was 16.9 months (SD ± 7.2; range 10-24). The auditory skills were evaluated with the Infant Toddler Meaningful Inventory Scale and the Category of Auditory Performance. Lexical level was assessed with the MacArthur-Bates Communicative Development Inventory. The overall data of average daily use and acoustic scene-analyses were extracted from Data Logging system. The effect of the one-year cumulative listening time to speech (in quiet) and speech-in-noise on the auditory and lexical scores was analysed. A significant positive correlation was found between speech in quiet exposure time at low loudness level (<70 dB) and lexical quotient after one year of CI use. Infant Toddler Meaningful Inventory Scale was negatively correlated with the highest speech-in-noise loudness levels (>80 dB). The Category of Auditory Performance was not related to the logged data. The listening environment can influence the early functional outcomes in younger implanted children. In this perspective, the data logging system is a promising tool in predicting early linguistic and auditory outcomes.


Occurrence of ochratoxin A and B in spices.
A total of 681 samples of spices, which comprised more than 50 different spice commodities were analysed for the natural occurrence of the mycotoxins ochratoxin A (OTA) and ochratoxin B (OTB). The analytical method involved chloroform extraction, clean-up by immunoaffinity column and HPLC determination of both mycotoxins. OTA and OTB were detected in 143 (21%) and 68 (10%) of the samples, respectively. The highest frequency of occurrence of both mycotoxins detected were in chili (100% for OTA and 55% for OTB), paprika (41% and 15%, respectively) and pepper (23% and 44%, respectively).The toxin concentrations ranged between the detection limit (0.01 ng/g) and 41.8 ng OTA (2.7 ng OTB)/g of chili, 18.9 ng OTA (1.4 ng OTB)/g of paprika and 3.8 ng OTA (4.6 ng OTB)/g of pepper. One sample of a extract of vanilla was found to be positive for OTB at 15 ng/g. However, median values of most samples showed to be below the detection limit. Comparison of the geographical origin of the samples showed that the predominant number of contaminated spices was from Southeast-Asia and India. Highly contaminated paprika samples were found to come from Israel.


Molecular detection of Anaplasma phagocytophilum and Borrelia burgdorferi sensu lato genospecies in red foxes (Vulpes vulpes) from Romania.
Red foxes (Vulpes vulpes) are one of the most widespread wild carnivores in the world, being recognized to harbor and transmit a wide range of vector-borne diseases. Anaplasma phagocytophilum and Borrelia burgdorferi sensu lato are zoonotic tick-borne pathogens causing emerging diseases. Wild animals play an essential role in the transmission of diseases and pathogens maintenance in nature. Epidemiological studies regarding the prevalence of tick-borne pathogens in red foxes are of public health importance, as they may successfully act as a pathogen transmission interface between wildlife, domestic animals and humans. This study included 14 counties from Romania. A total number of 353 red foxes (Vulpes vulpes) were examined. Heart tissue samples were collected during necropsy and stored at -20 °C. Genomic DNA extraction was performed and all samples were examined by polymerase chain reaction (PCR). Specific primers for A. phagocytophilum, A. platys, E. canis and Borrelia burgdorferi s.l. were used. Sequence analysis was performed (Macrogen Europe, Amsterdam) and obtained sequences are available at GenBank™. Out of the 353 samples, 9 (2.55 %; 95 % CI: 1.25-4.96 %) were positive for A. phagocytophilum. Positive animals originated from 5 counties. In total, 5 out of 353 heart tissue samples (1.42 %; 95 % CI: 0.52-3.47 %) collected from red foxes were positive for B. burgdorferi s.l. Red foxes originated from 4 counties. None of the samples were positive for A. platys or E. canis. No co-infection with A. phagocytophilum and B. burgdorferi s.l. was found. This first report of A. phagocytophilum and B. burgdorferi s.l. in red foxes from Romania suggests a limited role of foxes in the maintenance of the two related pathogens, but may represent a potential risk from a public health perspective.


Effect of acidic, neutral and alkaline conditions on product distribution and biocrude oil chemistry from hydrothermal liquefaction of microalgae.
Hydrothermal liquefaction (HTL) of microalgae produces high amount of water-insoluble organic compounds, the biocrude oil. Using high-growth-rate Spirulina platensis as feedstock, product fraction distribution and biocrude oil chemistry from HTL at a temperature of 240-300 °C under acidic, neutral and alkaline condition were studied. Positive effects on biocrude oil yield were only found with KOH and acetic acid, and these effects were stronger under milder HTL conditions. FT-ICR MS showed that O2 class in the biocrude was high due to higher carbohydrate in the biomass, numbers of N3O5-6 species present in the sample from acetic acid run, indicating its less decarboxylation ability. GC-MS showed more ketones and amides were formed from fatty acids in catalytic HTL, and this effect was sensitive toward reaction temperature. GPC suggested more light volatiles were in biocrude from KOH run, while analysis from NMR, FT-IR and elemental confirmed its high oil quality.


Late effects of childhood ALL treatment on body mass index and serum leptin levels.
It is well known that survivors of acute lymphoblastic leukemia (ALL) show a tendency to become overweight. Cranial irradiation (CRT), is considered to be the primary risk factor for development of obesity. The aim of our study was to evaluate body mass index (BMI) and serum leptin levels in survivors of childhood ALL. Subjects (Group I) consisted of 93 survivors of childhood ALL (53 males > or = 9 years old, 40 females > or = 8 years old) diagnosed between January 1975 and December 2002 in the Hematology-Oncology Division in Cerrahpasa Medical Faculty, Istanbul University after a follow up 10.21 +/- 4.90 (mean +/- SD) years. Fifty healthy individuals of similar chronological age were taken as controls (29 males, 21 females). Seventy-four subjects had received radiotherapy (Group IA) and 19 had not (Group I B). In Group I, BMI was significantly higher than in Group II (21.65 +/- 4.02 vs 20.31 +/- 3.49, p = 0.04). However, BMI was significantly higher only in Group I A (21.83 +/- 4.27) than in Group II (p = 0.032). Leptin levels were significantly higher in Group I A females than in Group II females. There was a significant correlation between BMI-SDS and serum leptin levels in group IA females. Leukemia treatment leads to obesity. Higher leptin levels in girls may suggest that sex may be a differentiating factor for this late effect.


A3B type unsymmetrical and amphiphilic phthalocyanines: Synthesis, characterization, thermal stability and aggregation studies.
This study reports the preparation, analysis, aggregation statuses and thermal properties of five phthalocyanines. The preparation of all molecules includes two steps: the first step is the preparation of phthalocyanine precursor molecules (A and B coded phthalonitriles); the second step is the synthesis of one metal-free and four metalled phthalocyanines (A3B-Co, A3B-Cu, A3B-H, A3B-Ni, and A3B-Zn coded compounds). Compound A holds a hydrophobic chain (cetyl alcohol); compound B holds a hydrophilic chain (tetraethylene glycol). Phthalocyanines include three hydrophobic chains and one hydrophilic chain (A3B type phthalocyanine). Characterization methods which were used to determine the structure of these compounds are Fourier Transform Infrared (FTIR), Proton Nuclear Magnetic Resonance (1H NMR), Carbon-13 Nuclear Magnetic Resonance (13C NMR), and Ultraviolet Visible (UV Vis) spectroscopies and elemental analysis. The phthalocyanines have high stability up to 260 °C as a minimum value. Aggregation statuses of the phthalocyanines change to the metal from the metal or to the solvent from the solvent.


Metastatic patterns and survival outcomes in patients with stage IV colon cancer: A population-based analysis.
The goal of this study was to delineate the patterns of distant metastasis from colon adenocarcinoma (CAC) and evaluate the survival differences by metastatic patterns. Using the Surveillance, Epidemiology, and End Results (SEER) database, we extracted patients diagnosed with stage IV CAC between 2010 and 2016. Kaplan-Meier survival curves were plotted with log-rank tests to compare overall survival (OS) of patients with different metastatic patterns. Univariate and multivariate Cox proportional hazards regression models were used to evaluate the effects of different metastatic patterns on survival outcomes in terms of OS and disease-specific survival (DSS). A total of 26 170 patients were analyzed. The 3- and 5-year OS were 20.7% and 10.5%, respectively, for patients with stage IV CAC. The most common distant metastatic site was the liver, followed by the lung, bone, and brain, but the frequency differed greatly by histology subtypes. The site of metastasis was a significant prognostic factor for OS and DSS in patients with stage IV CAC, independent of the number of metastatic sites and other clinical and demographic prognostic factors. Using liver-only metastasis as reference, lung-only metastasis was associated with better OS (hazard ratio [HR] = 0.82, 95% confidence interval [CI], 0.71-0.94) and DSS (HR = 0.75, 95% CI, 0.64-0.88). Older age, black race, unmarried status, grade III/IV tumors, advanced tumor-node-metastasis (TNM) stage, proximal colon, elevated preoperative carcinoembryonic antigen (CEA), no surgery of the primary site, and no chemotherapy were independent predictors of poor OS. The site of distant metastasis and number of metastasis site were independent prognostic factors for survival of patients with stage IV CAC. This study highlights the need for diverse treatment strategies for patients with different metastatic patterns.


What are the prevalence of and factors independently associated with depression and anxiety among patients with posttraumatic elbow stiffness? A cross-sectional, multicenter study.
Joint stiffness is a common complication after articular-related trauma in the elbow, resulting in significant limb disability, psychological stress, and a negative impact on daily life. No previous study has reported the impact of post-traumatic elbow stiffness (PTES) on psychological health. This study aims to (1) investigate the depression and anxiety levels and (2) identify factors independently associated with depression and anxiety symptoms in patients with PTES. A total of 108 patients with PTES presenting to 4 collaborative municipal hospitals were consecutively enrolled from September to December 2020. Sociodemographic and clinical characteristics were collected through questionnaires and medical records. The Depression Anxiety Stress Scale-21 was used to assess depression and anxiety status. Ordinal logistic regression analysis was performed to identify factors independently associated with depression and anxiety symptoms. The detection rates of mild-to-moderate depression and anxiety are 40.7% and 27.8%, and severe-to-extremely severe levels are 23.1% and 25.9%, respectively. Regression results show that factors independently associated with depression include elbow flexion (odds ratio [OR]per 1° loss = 1.021, 95% confidence interval [CI]: 1.001-1.041, P = .035), elbow pain on movement (ORper 1 point increase = 1.236, 95% CI: 1.029-1.484, P = .023), family relationship (ORless close/very close = 10.059, 95% CI: 2.170-46.633, P = .003), and self-care ability (ORunable/able = 3.858, 95% CI: 1.244-11.961, P = .019). Factors independently associated with anxiety are elbow flexion (ORper 1° loss = 1.031, 95% CI: 1.009-1.052, P = .005), elbow pain on movement (ORper 1 point increase = 1.212, 95% CI: 1.003-1.465, P = .047), and clinically significant heterotopic ossification around elbow (ORyes/no = 2.344, 95% CI: 1.048-5.243, P = .038). Patients with PTES exhibit significant depression and anxiety symptoms. Several sociodemographic and clinical characteristics are independently associated with depression and anxiety levels. Identifying and addressing these factors may be of particular benefit during PTES management. Future research might address whether depression and anxiety affect the outcome after stiff elbow surgery.


Crystal structure and characterization of a novel L-serine ammonia-lyase from Rhizomucor miehei.
L-serine ammonia-lyase, as a member of the β-family of pyridoxal-5'-phosphate (PLP) dependent enzymes, catalyzes the conversion of L-serine (L-threonine) to pyruvate (α-ketobutyrate) and ammonia. The crystal structure of L-serine ammonia-lyase from Rhizomucor miehei (RmSDH) was solved at 1.76 Å resolution by X-ray diffraction method. The overall structure of RmSDH had the characteristic β-family PLP dependent enzyme fold. It consisted of two distinct domains, both of which show the typical open twisted α/β structure. A PLP cofactor was located in the crevice between the two domains, which was attached to Lys52 by a Schiff-base linkage. Unique residue substitutions (Gly78, Pro79, Ser146, Ser147 and Thr312) were discovered at the catalytic site of RmSDH by comparison of structures of RmSDH and other reported eukaryotic L-serine ammonia-lyases. Optimal pH and temperature of the purified RmSDH were 7.5 and 40 °C, respectively. It was stable in the pH range of 7.0-9.0 and at temperatures below 40 °C. This is the first crystal structure of a fungal L-serine ammonia-lyase. It will be useful to study the catalytic mechanism of β-elimination enzymes and will provide a basis for further enzyme engineering.


Metabolic effects of glucagon in endogenous hypertriglyceridemia.
The metabolic effects of glucagon, administered i.v. in doses of 1 microgram/kg, were evaluated in two groups of patients with endogenous hypertriglyceridemia (Types IV and V according to Fredrickson) with normal and reduced glucose tolerance and in a control group. Glucagon had a lipolytic effect, evaluated as the plasma increase of free fatty acids (FFA) during the first 20 min in normal subjects, but not in the two hyperlipemic groups. A negative correlation was observed between fasting IRI level and FFA mobilization. The ketogenic and hypotriglyceridemic effects of glucagon were demonstrated in normal and hyperlipemic groups. It would seem, therefore, that at the pharmacological doses injected, there is no resistance to the hypotriglyceridemic effect of glucagon in endogenous hypertriglyceridemia.


Hemodynamic assessment and management of patients with the adult respiratory distress syndrome.
In summary, ARDS and the combination of MV and PEEP may have complicated hemodynamic effects in terms of cardiac output and regional blood flow. These effects should be assessed primarily in terms of the adequacy of tissue oxygenation. Hemodynamic management of patients with ARDS should be aimed at improving tissue oxygenation by reducing metabolic needs, and improving both the arterial oxygen content (Table 3) and the cardiac output (Table 4). In most patients, these goals can be realized simultaneously.


Clinical progression of moderate-to-severe Alzheimer's disease and caregiver burden: a 12-month multicenter prospective observational study.
Prospective studies on the clinical progression of Alzheimer's disease (AD) and its relationship to caregiver burden are needed to improve illness management and use of resources. This national, multicenter, observational study evaluated 1235 moderate to severe AD patients under routine care in Spain. Baseline cross-sectional sociodemographic and clinical data, and changes from baseline to month 12 of various neuropsychological tests and clinical ratings, including Blessed Dementia Scale, Mini-mental State Examination (MMSE), Hughes Clinical Dementia Rating sum-of-boxes (CDR-SB), Clinical Global Impression of Change (CGIC) and Zarit Caregiver Burden scales, were recorded and comprehensively analyzed. Baseline data were in accordance with characteristics consistently reported to influence AD risk regarding anthropometrics, sociocultural features and comorbidities. Significant progressive functional impairments (i.e. in routine activities and essential daily tasks) and cognitive (i.e. MMSE and CDR-SB) impairments were found at month 12. However, patients' behavior and caregivers' burden improved slightly, but significantly, corroborating the major influence of behavioral symptoms on caregivers' distress. Caregivers showed significantly lower burden with patients with higher levels of education and, to a lesser extent, when patients received AD-specific medication. Physicians accurately detected AD clinical evolution as their CGIC ratings significantly correlated with all tests. These findings reinforce previous AD knowledge and add data on the clinical course of advanced stages of AD. Caregiver burden depended more on patients' behavioral alterations than on their functional or cognitive declines; and it was diminished by their patients having higher levels of education and being treated with AD-specific medications. Research into unexplored factors that might reduce caregiver burden, ultimately benefiting both patients and caregivers, is encouraged.


PTHrP and breast cancer: more than hypercalcemia and bone metastases.
Parathyroid hormone-related protein (PTHrP) causes hypercalcemia in cancer patients. PTHrP is required for normal breast development and has been shown to promote bone metastases from breast cancers. However, whether the protein also contributes to the formation of primary tumors has been unclear. Two recent papers suggest it may. First, a report in Nature Genetics identified the PTHrP locus as a new breast cancer susceptibility gene. Second, a paper in Journal of Clinical Investigation demonstrated that PTHrP promotes tumor growth and metastases in MMTV-PyMT mice. These studies implicate PTHrP in the development and growth of primary breast tumors and underscore the need for further research.


Types of private speech produced by hyperactive and nonhyperactive boys.
Types and amount of private speech (audible talking that is not addressed to another person) were assessed during the free play of 16 hyperactive and 16 nonhyperactive boys. Verbalizations were coded into nine categories that denoted the boys' level of use of verbal control of their own behavior (Luria, 1961; Kohlberg, Yeager, & Hjertholm; 1968). Differences in amount and type of private speech between hyperactive and nonhyperactive boys were found to indicate that hyperactive boys may be presenting a specific or general cognitive lag in development. Treatment ramifications are discussed.


Patients' perception of colorectal cancer surveillance in the community: an exploratory study.
All patients who underwent curative resection for colorectal cancer (CRC) are frequently reviewed in tertiary institutions to ensure timely detection of any disease recurrence. There has been no local study that evaluated the feasibility of monitoring their condition in the community as a possible new model of care. This study henceforth seeks to understand CRC patients' views and receptiveness of having their surveillance consultations conducted in a community setting. We convenience sampled Stage I and II CRC patients who were within five years post-operation in the outpatient clinics. An open-ended questionnaire aimed at elucidating their perception towards cancer surveillance in a community setting was administered. Content analysis was used to group and quantify responses from participants. Twenty-five participants agreed to participate in the study. Only 48% of the participants felt that having phlebotomy procedures in a community or home setting was acceptable. Participants were less willing to be reviewed by a physician who is not their primary surgeon, with only 32% agreeable to seeing a different doctor for surveillance if given a choice. However, most participants were open to having a telephone consultation in place of a physical face-to-face consultation before (72%) and after (76%) going through medical imaging. Participants remained keen to be managed by their primary surgeons and were hesitant towards having their follow-up surveillance consultations in community and primary care settings. Further studies should be conducted to understand whether these perceptions are generalisable, and if more can be done to change public perception towards the role of community and primary care institutions.


Adapalene 0.1%/benzoyl peroxide 2.5% gel: a review of its use in the treatment of acne vulgaris in patients aged ≥ 12 years.
Adapalene 0.1%/benzoyl peroxide 2.5% gel (Epiduo™, Tactuo™) is the only fixed-dose combination product available that combines a topical retinoid with benzoyl peroxide; it targets three of the four main pathophysiologic factors in acne. This article reviews the therapeutic efficacy and tolerability of topical adapalene 0.1%/benzoyl peroxide 2.5% gel in the treatment of patients aged ≥ 12 years with acne vulgaris, as well as summarizing its pharmacologic properties. In three 12-week trials in patients aged ≥ 12 years with moderate acne, success rates were significantly higher with adapalene 0.1%/benzoyl peroxide 2.5% gel than with adapalene 0.1% gel or benzoyl peroxide 2.5% gel alone, and combination therapy had an earlier onset of action. In addition, significantly greater reductions in total, inflammatory, and noninflammatory lesion counts were seen in patients receiving adapalene 0.1%/benzoyl peroxide 2.5% gel than in those receiving adapalene 0.1% gel or benzoyl peroxide 2.5% gel alone. Adapalene 0.1%/benzoyl peroxide 2.5% gel did not significantly differ from clindamycin 1%/benzoyl peroxide 5% gel in terms of the reduction in the inflammatory, noninflammatory, or total lesion counts in patients with mild to moderate acne, according to the results of a 12-week trial. Twelve-week studies showed that topical adapalene 0.1%/benzoyl peroxide 2.5% gel in combination with oral lymecycline was more effective than oral lymecycline alone in patients with moderate to severe acne, and topical adapalene 0.1%/benzoyl peroxide 2.5% gel in combination with oral doxycycline hyclate was more effective than oral doxycycline hyclate alone in patients with severe acne. In patients with severe acne who responded to 12 weeks' therapy with topical adapalene 0.1%/benzoyl peroxide 2.5% gel plus oral doxycycline hyclate or oral doxycycline hyclate alone, an additional 6 months' therapy with adapalene 0.1%/benzoyl peroxide 2.5% gel was more effective than vehicle gel at maintaining response, with further improvement seen in adapalene 0.1%/benzoyl peroxide 2.5% gel recipients. A noncomparative study also demonstrated the efficacy of 12 months' therapy with adapalene 0.1%/benzoyl peroxide 2.5% gel in patients with acne vulgaris. Topical adapalene 0.1%/benzoyl peroxide 2.5% gel was generally well tolerated in patients with acne. In 12-week trials, the most commonly occurring treatment-related adverse events included erythema, scaling, dryness, and stinging/burning; these dermatologic treatment-related adverse events were usually of mild to moderate severity, occurred early in the course of treatment, and resolved without residual effects. Topical adapalene 0.1%/benzoyl peroxide 2.5% gel was generally well tolerated in the longer term, with dry skin being the most commonly occurring treatment-related adverse event over 12 months of treatment. In conclusion, adapalene 0.1%/benzoyl peroxide 2.5% gel is a valuable agent for the first-line treatment of acne vulgaris.


Prediction and Reduction of the Aggregation of Monoclonal Antibodies.
Protein aggregation remains a major area of focus in the production of monoclonal antibodies. Improving the intrinsic properties of antibodies can improve manufacturability, attrition rates, safety, formulation, titers, immunogenicity, and solubility. Here, we explore the potential of predicting and reducing the aggregation propensity of monoclonal antibodies, based on the identification of aggregation-prone regions and their contribution to the thermodynamic stability of the protein. Although aggregation-prone regions are thought to occur in the antigen binding region to drive hydrophobic binding with antigen, we were able to rationally design variants that display a marked decrease in aggregation propensity while retaining antigen binding through the introduction of artificial aggregation gatekeeper residues. The reduction in aggregation propensity was accompanied by an increase in expression titer, showing that reducing protein aggregation is beneficial throughout the development process. The data presented show that this approach can significantly reduce liabilities in novel therapeutic antibodies and proteins, leading to a more efficient path to clinical studies.


Efficacy of Reslizumab Treatment in Exacerbation-Prone Patients with Severe Eosinophilic Asthma.
Patients with severe eosinophilic asthma have increased risk of clinical asthma exacerbations (CAEs), impaired lung function, and lower quality of life (QoL) compared with patients with noneosinophilic asthma. The efficacy and safety of intravenous reslizumab have been demonstrated in 2 duplicate, randomized, double-blind, placebo-controlled, phase 3 studies. We present findings from post hoc analyses of the subgroup of patients from the phase 3 studies with severe (Global Initiative for Asthma [GINA] Step 4 or 5) eosinophilic asthma who had ≥2 or ≥3 CAEs in the 12 months before screening. Patients aged ≥12 years with severe eosinophilic asthma were randomized (1:1) to reslizumab 3.0 mg/kg or placebo every 4 weeks for 1 year. Outcomes assessed included CAEs, forced expiratory volume in 1 second (FEV1), and Asthma Control Questionnaire 6 (ACQ-6) and Asthma QoL Questionnaire (AQLQ) scores. Of 953 patients randomized, 318 (reslizumab: n = 156; placebo: n = 162) and 164 (reslizumab: n = 72; placebo: n = 92) were GINA 4/5 with ≥2 CAEs in the prior year and ≥3 CAEs in the prior year, respectively. Reslizumab significantly improved CAE rate, time to first CAE, and the proportion of patients who experienced ≥1 CAE versus placebo in both CAE subgroups. Improvements in FEV1, ACQ-6 and AQLQ scores, and systemic corticosteroid burden were also observed with reslizumab versus placebo after 52 weeks of treatment in both CAE subgroups. Reslizumab reduced CAE frequency and improved lung function, asthma control, and QoL versus placebo in patients with severe eosinophilic asthma with a high CAE rate before treatment.


Role of quasiresonant planetary wave dynamics in recent boreal spring-to-autumn extreme events.
In boreal spring-to-autumn (May-to-September) 2012 and 2013, the Northern Hemisphere (NH) has experienced a large number of severe midlatitude regional weather extremes. Here we show that a considerable part of these extremes were accompanied by highly magnified quasistationary midlatitude planetary waves with zonal wave numbers m = 6, 7, and 8. We further show that resonance conditions for these planetary waves were, in many cases, present before the onset of high-amplitude wave events, with a lead time up to 2 wk, suggesting that quasiresonant amplification (QRA) of these waves had occurred. Our results support earlier findings of an important role of the QRA mechanism in amplifying planetary waves, favoring recent NH weather extremes.


Benefit to N2-fixing alder of extending growth period at the cost of leaf nitrogen loss without resorption.
This study examines the adaptive role of not resorbing N in N(2)-fixing deciduous trees in terms of their energy balance. The autumnal growth of N(2)-fixing Alnus firma Sieb. et Zucc. (alder) was compared with that of the non-N(2)-fixing Morus bombycis Koizumi (mulberry), which resorbs leaf N. The freezing resistance of leaves of both species was -2 degrees C. Mulberry seedlings lost their photosynthetic ability in mid-October, although the minimum temperature was still above 0 degrees C. Thereafter, their leaves turned yellow and were gradually shed. In contrast, seedlings of the alder maintained their photosynthetic ability until mid-November, when the minimum temperature fell to the freezing resistance limit. Thereafter, their leaves were shed quickly without an autumn tint. The mulberry resorbed 48.9% of leaf N, whereas the alder resorbed hardly any. These results show that, compared with the mulberry tree, the alder extended its growth period for 1 month in return for losing leaf N without resorption. The amount of energy assimilated by the alder in the extended growth period was about six times that required for compensating for the nitrogen loss, if the compensation is dependent only on the tree's own nitrogen fixation. This surplus energy balance has probably allowed N(2)-fixing deciduous trees to evolve their non-N-resorbing trait.


The development of tone in the smooth muscle of guinea-pig isolated tracheal preparations may be influenced by prostanoids released from the adjacent airway cartilage.
Guinea-pig tracheal strip preparations containing cartilage, placed under an applied load in vitro, develop tone spontaneously. The finding that spontaneous tone is reduced by indomethacin suggests that one or more prostanoids are involved in the development of spontaneous tone in this species. In this study we examined the effects of removing the cartilage component of the preparations on changes in tone induced by indomethacin and isoproterenol. In contrast to preparations containing cartilage, tissues devoid of cartilage, did not develop tone after the application of an initial 1 g resting load. Indomethacin (1 microM) reduced resting tone by 0.62 +/- 0.14 g in cartilage-containing tissues but, in contrast, reduced tone by only 0.03 +/- 0.01 g in tissues devoid of cartilage. Furthermore, relaxation responses (0.38 +/- 0.05 g) to isoproterenol (1 microM) could be produced in cartilage-containing preparations but not in cartilage-free preparations. Radioimmunoassays indicated that the release of PGE2, PGF2 alpha and 6-keto PGF1 alpha, the end-product of PGI2 breakdown, was diminished in preparations lacking cartilage. Thus, in guinea-pig airway preparations cartilage is apparently a source of sufficient prostanoids to induce spontaneous tone.


Rapid changes in the lymphopoietic and granulopoietic compartments of the marrow caused by stress levels of corticosterone.
Exposure to concentrations of glucocorticoids analogous to those produced during stress, trauma and malnutrition had rapid but varying effects on the major classes of cells within the marrow. Corticosterone (CS) was given as a subdermal implant in young mice and generated 60-95 microg CS/dl of blood compared to 5-15 microg CS/dl for sham controls over a period of 36 hr. Within 24 hr CS had caused losses of 30-70% among the early pro-B, pre-B and immature B cells. The pre-B cells were virtually eliminated by 36 hr and the capacity of surviving pro- and pre-B cells to cycle was reduced by 70-80%. Interestingly, the earliest of B cells, the prepro-B cells, showed considerable resistance to CS, being reduced by only 20% at 36 hr. Thus, the pattern of survival within the B-cell compartment paralleled the expression of Bcl-2. At the 36-hr time-point there were no changes in the proportion of progenitor cells, erythroid or monocytic cells, or number of nucleated cells in the marrow. By contrast, 36 hr after exposure to CS there was an increase of 30% in the proportion and absolute number of cells in the granulocytic compartment. Chronic production of CS appears to reprogramme lymphopoiesis and myelopoiesis, perhaps to preserve the first line of immune defence at the expense of the lymphoid branch. Resistance to apoptosis and modifications in the activity of the glucocorticoid receptor and cytokines produced by stromal cells are postulated as targets for CS-driven changes.


Morphometric Analysis of the External Auditory Canal by Computed Tomography in Indian Population.
Various studies have shown variations in size and shape of different anthropometric measurements of external auditory canal. We conducted an anthropometric study of the three-dimensional anatomy of the osseous external auditory canal (OEAC) using high-resolution computed tomography the temporal bone to identify the variations in subset of Indian population from North India. A retrospective review of high-resolution computed tomography images of the temporal bones of 125 patients (250 external auditory canals) of different ages (mean 28.43 years) acquired from September 2014 to February of 2015 was performed. Using a method, as proposed by Mahboubio et al. (Otol Neurotol 33:715-720, 2012), six defined dimensions of the OEAC in the parasagittal planes were recorded at the level of annulus, midcanal and the outermost point of osseous external auditory canal at bony-cartilaginous junction. The length and shape of the OEAC were also studied and the frequency rate of each was recorded. The most prevalent shape of the OEAC was found to be conical (64%) and the mean osseous external auditory canal length was 9.61 mm. The length of the OEAC was significantly different between ages above and below 12 years while the 6 defined cross sectional dimensions were statistically significant between ages above and below 8 years. The history of chronic suppurative otitis media had a significant bearing on the inferior mid-anteroposterior dimension at the level of bony-cartilaginous junction. There was statistically significant difference in supero-inferior diameter in the posterior half at the level of mid-canal and outer bony-cartilaginous junction between males and females. The comprehensive set of standardized measurements collected in this study provides three-dimensional information on osseous external auditory canal geometry. These measurements and the methodology will contribute to the development of element models of the osseous canal for computational modeling purposes and also provide important measurements for design of in-the-canal hearing aids, specialized earplugs and for defining average sizes for canalplasty procedures, in pre- and postoperative surgical planning and assessment of canal atresia and stenosis in Indian population. No such previous study has been done in North Indian population.


Hemodynamic effects of amrinone and colloid administration in children following cardiac surgery.
Amrinone was used as the sole vasoactive medication in 9 of 14 children (aged 5 months to 8.25 years) given the drug following open repair of congenital cardiac lesions. Four children received a concomitant dopamine infusion and one infant had the infusion stopped after 5 hours for low mean arterial pressure (49 mmHg). In the 10 children receiving only amrinone, cardiac index increased 21% (range, 0 to 94%) after a total loading dose of 4.5 mg/kg given over 1 hour. Four of 14 patients (29%) required dopamine infusions to maintain mean arterial pressure over 55 mmHg and in these children cardiac index increased from baseline and was maintained during the amrinone infusion. Preload was held constant by administration of whole blood or plasmanate during amrinone loading; a decrease in systemic vascular resistance index was seen resulting in a stable arterial blood pressure. Minimal chronotropic effect was seen and no arrhythmias occurred. The sole child with postoperative pulmonary hypertension had a beneficial decrease in pulmonary artery pressure, increase in cardiac index, and stable systemic blood pressure during amrinone use. Cardiac index changes during amrinone loading in these children were variable and less clearly related to serum levels than reported in adults. Pharmacokinetic analysis in 12 children showed a clearance of 3.4 mL/min/kg, a volume of distribution of 1.65 L/kg, and an elimination half-life of 5.75 hours. Decreases in platelet counts were seen in 6 children and platelet transfusion was needed in 1; thus, serial platelet counts should be monitored.(ABSTRACT TRUNCATED AT 250 WORDS)


Protective effect of ginsenoside Rd on military aviation noise-induced cochlear hair cell damage in guinea pigs.
Noise pollution has become one of the important social hazards that endanger the auditory system of residents, causing noise-induced hearing loss (NIHL). Oxidative stress has a significant role in the pathogenesis of NIHL, in which the silent information regulator 1(SIRT1)/proliferator-activated receptor-gamma coactivator 1α (PGC-1α) signaling pathway is closely engaged. Ginsenoside Rd (GSRd), a main monomer extract from ginseng plants, has been confirmed to suppress oxidative stress. Therefore, the hypothesis that GSRd may attenuate noise-induced cochlear hair cell loss seemed promising. Forty-eight male guinea pigs were randomly divided into four groups: control, noise exposure, GSRd treatment (30 mg/kg Rd for 10d + noise), and experimental control (30 mg/kg glycerol + noise). The experimental groups received military helicopter noise exposure at 115 dB (A) for 4 h daily for five consecutive days. Hair cell damage was evaluated by using inner ear basilar membrane preparation and scanning electron microscopy. Terminal dUTP nick end labeling (TUNEL) and immunofluorescence staining were conducted. Changes in the SIRT1/PGC-1α signaling pathway and other apoptosis-related markers in the cochleae, as well as oxidative stress parameters, were used as readouts. Loss of outer hair cells, more disordered cilia, prominent apoptosis, and elevated free radical levels were observed in the experimental groups. GSRd treatment markedly mitigated hearing threshold shifts, ameliorated outer hair cell loss and lodging or loss of cilia, and improved apoptosis through decreasing Bcl-2 associated X protein (Bax) expression and increasing Bcl-2 expression. In addition, GSRd alleviated the noise-induced cochlear redox injury by upregulating superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) levels, decreasing malondialdehyde (MDA) levels, and enhancing the activity of SIRT1 and PGC-1α messenger ribonucleic acid (mRNA) and protein expression. In conclusion, GSRd can improve structural and oxidative damage to the cochleae caused by noise. The underlying mechanisms may be associated with the SIRT1/PGC-1α signaling pathway.


MicroRNA-487b Is a Negative Regulator of Macrophage Activation by Targeting IL-33 Production.
MicroRNAs (miRNAs) are short noncoding RNAs that regulate a broad spectrum of biological processes, including immune responses. Although the contributions of miRNAs to the function of immune cells are beginning to emerge, their specific roles remain largely unknown. IL-33 plays an important role in macrophage activation for innate host defense and proinflammatory responses. In this study, we report that miR-487b can suppress the levels of mRNA and protein for IL-33 during the differentiation of bone marrow-derived macrophages (BMDMs). This results in inhibition of IL-33-induced expression of Ag-presenting and costimulatory molecules and proinflammatory mediators. A luciferase assay showed that miR-487b binds to the IL-33 3'-untranslated region. We also confirmed that IL-33 directly promotes the activation of BMDMs by increasing the expression of MHC class I, MHC class II, CD80/CD86, and inducible NO synthase (iNOS) in a dose-dependent manner. Exposure of BMDMs to the TLR4 ligand, LPS, decreased miR-487b expression, increased IL-33 transcript levels, and induced the production of proinflammatory mediators (e.g., iNOS, IL-1β, IL-6, and TNF-α). Treatment with a specific inhibitor of miR-487b function also resulted in increased levels of IL-33 mRNA, which augmented LPS-induced expression of these inflammatory mediators in macrophages. Collectively, our results indicate that miR-487b plays a negative regulatory role in macrophages by controlling the levels of IL-33 transcript and protein to fine-tune innate immune host defense and proinflammatory responses of these cells. Thus, miR-487b plays an important role in the regulation of macrophage homeostasis and activation by targeting IL-33 transcripts.


Skewed X-chromosome inactivation plays a crucial role in the onset of symptoms in carriers of Becker muscular dystrophy.
Becker muscular dystrophy (BMD) is an X-linked recessive disorder affecting approximately 1: 18.000 male births. Female carriers are usually asymptomatic, although 2.5-18% may present muscle or heart symptoms. In the present study, the role of the X chromosome inactivation (XCI) on the onset of symptoms in BMD carriers was analysed and compared with the pattern observed in Duchenne muscular dystrophy (DMD) carriers. XCI was determined on the lymphocytes of 36 BMD carriers (both symptomatic and not symptomatic) from 11 families requiring genetic advice at the Cardiomyology and Medical Genetics of the Second University of Naples, using the AR methylation-based assay. Carriers were subdivided into two groups, according to age above or below 50 years. Seven females from the same families known as noncarriers were used as controls. A Student's t-test for nonpaired data was performed to evaluate the differences observed in the XCI values between asymptomatic and symptomatic carriers, and carriers aged above or below 50 years. A Pearson correlation test was used to evaluate the inheritance of the XCI pattern in 19 mother-daughter pairs. The results showed that symptomatic BMD carriers had a skewed XCI with a preferential inactivation of the X chromosome carrying the normal allele, whereas the asymptomatic carriers and controls showed a random XCI. No concordance concerning the XCI pattern was observed between mothers and related daughters. The data obtained in the present study suggest that the onset of symptoms in BMD carriers is related to a skewed XCI, as observed in DMD carriers. Furthermore, they showed no concordance in the XCI pattern inheritance.


Antigenic reactivity of the major glycoprotein of equine infectious anemia virus, a retrovirus.
The immunogenic contributions of the carbohydrate and peptide portions of the major envelope glycoprotein of equine infections anemia virus, EIAV gp90, were analyzed by measuring the effects of specific glycosidase and protease digestions on the reactivity of the glycoprotein with immune sera from infected horses. The results of both direct and competitive radioimmunoassay demonstrated that immune sera contained antibodies reactive with both the carbohydrate and protein moieties of EIAV gp90, with the predominant reactivity apparently against the gp90 peptide epitopes. These results contrast with previous descriptions of retrovirus glycoprotein antigenicity in which antigenic reactivity was attributed exclusively to protein (Friend murine leukemia virus) or to carbohydrate (bovine leukemia virus). Thus EIAV gp90 displays a complex antigenicity which must be taken into consideration in analyzing the biochemical nature of antigenic variation of the virus during persistent infections in horses.


Accuracy and ease-of-use of seven point-of-care SARS-CoV-2 antigen-detecting tests: A multi-centre clinical evaluation.
Antigen-detecting rapid diagnostic tests (Ag-RDTs) for SARS-CoV-2 are important diagnostic tools. We assessed clinical performance and ease-of-use of seven Ag-RDTs in a prospective, manufacturer-independent, multi-centre cross-sectional diagnostic accuracy study to inform global decision makers. Unvaccinated participants suspected of a first SARS-CoV-2 infection were recruited at six sites (Germany, Brazil). Ag-RDTs were evaluated sequentially, with collection of paired swabs for routine reverse transcription polymerase chain reaction (RT-PCR) testing and Ag-RDT testing. Performance was compared to RT-PCR overall and in sub-group analyses (viral load, symptoms, symptoms duration). To understandusability a System Usability Scale (SUS) questionnaire and ease-of-use (EoU) assessment were performed. 7471 participants were included in the analysis. Sensitivities across Ag-RDTs ranged from 70·4%-90·1%, specificities were above 97·2% for all Ag-RDTs but one (93·1%).Ag-RDTs, Mologic, Bionote, Standard Q, showed diagnostic accuracy in line with WHO targets (> 80% sensitivity, > 97% specificity). All tests showed high sensitivity in the first three days after symptom onset (≥87·1%) and in individuals with viral loads≥ 6 log10SARS-CoV2 RNA copies/mL (≥ 88·7%). Usability varied, with Rapigen, Bionote and Standard Q reaching very good scores; 90, 88 and 84/100, respectively. Variability in test performance is partially explained by variable viral loads in population evaluated over the course of the pandemic. All Ag-RDTs reach high sensitivity early in the disease and in individuals with high viral loads, supporting their role in identifying transmission relevant infections. For easy-to-use tests, performance shown will likely be maintained in routine implementation. Ministry of Science, Research and Arts, State of Baden-Wuerttemberg, Germany, internal funds from Heidelberg University Hospital, University Hospital Charité - Universitätsmedizin Berlin, UK Department of International Development, WHO, Unitaid.


Divergent reactivity of platinum(ii) and palladium(ii) methylperoxo complexes and the formation of an unusual hemi-aminal complex.
The 6,6''-diaminoterpyridine palladium(ii) methylperoxo complex eliminates methyl hydroperoxide and reacts with acetone to form a novel hemi-aminal palladium complex, whereas the analogous platinum(ii) complex generates formaldehyde and a platinum(ii) hydroxo complex.


Acute methamphetamine administration upregulates NGFI-B mRNA expression in the striatum: co-localization with c-Fos immunoreactivity.
In this study, the effects of acute methamphetamine administration on expression of the nuclear transcription factor NGFI-B mRNA and its co-localization with c-Fos immunoreactivity in the striatum were evaluated in animals receiving a single dose of methamphetamine (4 mg/kg) given at 2 or 6 h prior to perfusion. All animals received a daily saline injection for 6 days prior to methamphetamine treatment. We have found that, similar to c-fos activation, NGFI-B mRNA levels were significantly increased 2 h after methamphetamine treatment and returned to basal levels 6 h later. Induction of NGFI-B mRNA levels by methamphetamine was highest in central striatum as compared to the dorsomedial distribution pattern observed in control animals. After acute methamphetamine treatment, the distribution pattern of NGFI-B mRNA upregulation was very similar to that of methamphetamine induced c-Fos immunoreactivity. However, co-localization studies with c-Fos immunoreactivity showed that not all NGFI-B-positive cells contained c-Fos after methamphetamine treatment. Forty-five percent of all NGFI-B mRNA expressing neurons contained c-Fos immunoreactivity in the dorsomedial striatum as compared to 60% in central and 35% in ventrolateral striatum. This study provides a detailed description of the differential spatial and temporal modulation of NGFI-B and c-Fos expression in the striatum by acute methamphetamine treatment over time.


FOXO3 growth inhibition of colonic cells is dependent on intraepithelial lipid droplet density.
Forkhead transcription factor FOXO3 plays a critical role in suppressing tumor growth, in part, by increasing the cell cycle inhibitor p27kip1, and Foxo3 deficiency in mice results in marked colonic epithelial proliferation. Here, we show in Foxo3-deficient colonic epithelial cells a striking increase in intracytoplasmic lipid droplets (LDs), a dynamic organelle recently observed in human tumor tissue. Although the regulation and function of LDs in non-adipocytes is unclear, we hypothesize that the anti-proliferative effect of FOXO3 was dependent on lowering LD density, thus decreasing fuel energy in both normal and colon cancer cells. In mouse colonic tumors, we found an increased expression of LD coat protein PLIN2 compared with normal colonic epithelial cells. Stimulation of LD density in human colon cancer cells led to a PI3K-dependent loss of FOXO3 and a decrease in the negative regulator of lipid metabolism in Sirtuin6 (SIRT6). Foxo3 deficiency also led to a decrease in SIRT6, revealing the existence of LD and FOXO3 feedback regulation in colonic cells. In parallel, LD-dependent loss of FOXO3 led to its dissociation from the promoter and decreased expression of the cell cycle inhibitor p27kip1. Stimulation of LD density promoted proliferation in colon cancer cells, whereas silencing PLIN2 or overexpression of FOXO3 inhibited proliferation. Taken together, FOXO3 and LDs might serve as new targets for therapeutic intervention of colon cancer.


Effect of environmental stress of low pressure on tyrosine aminotransferase and phenylalanine 4-hydroxylase activities in the rat.
1. Tyrosine aminotransferase activity in the liver increased about fourfold after 9h, on exposure of rats to stress of low pressure. 2. The phenylalanine hydroxylase activity increased about 60% on exposure for 24h or more. 3. An environmental pressure decrease of about 0.033 MN/m2 is needed to increase the activity of tyrosine aminotransferase. 4. Adrenalectomy completely abolished the increase in activity of tyrosine aminotransferase obtained on exposure to low pressure. 5. Treatment with cycloheximide or actinomycin D prevented the increase in activity of tyrosine aminotransferase. 6. Treatment with cycloheximide at the early part of exposure to stress prevented the increase in activity of phenylalanine hydroxylase obtained after 24h.


Establishment of an immortalized stromal cell line derived from human Endometriotic lesion.
Endometriosis is a benign gynecological disease with obviously feature of estrogen-dependence and inflammatory response. The applications of primary endometriotic stromal cells in research of endometriosis are restricted for short life span, dedifferentiation of hormone and cytokine responsiveness. The objective of this study was to establish and characterize immortalized human endometriotic stromal cells (ihESCs). The endometriotic samples were from a patient with ovarian endometriosis and the primary endometriotic stromal cells were isolated from the endometriotic tissues. The primary cells were infected by lentivirus to establish telomerase reverse transcriptase (hTERT)-induced immortalized cells. Quantification of mRNA and proteins was examined by quantitative real-time polymerase chain reaction (qRT-PCR) and Western Blot. CCK-8 assay and EdU labeling assay were assigned to assess the growth of ihESCs. Karyotype assay was performed to detect the chromosomes of ihESCs. Colony formation assay and nude mouse tumorigenicity assay were used to evaluate colony-formation and tumorigenesis abilities. ihESCs continuously overexpressed hTERT via infection of lentivirus and significant extended the life span reaching 31 passages. The morphology, proliferation and karyotype of ihESCs remained unchanged. The expression of epithelial-mesenchymal transition (EMT) markers, estrogen-metabolizing proteins and estrogen/progesterone receptors (ERs and PRs) were unaltered. Furthermore, the treatment of estrogen increased the proliferation and EMT of ihESCs. Lipopolysaccharides (LPS) and IL-1β remarkably induced inflammatory response. The clonogenesis ability of ihESCs was consistent with primary cells, which were much lower than Ishikawa cells. In addition, nude mouse tumorigenicity assay demonstrated that ihESCs were unable to trigger tumor formation. This study established and characterized an immortalized endometriotic stromal cell line that exhibited longer life span and kept the cellular morphology and physiological function as the primary cells. The immortalized cells remained normal feedback to estrogen and inflammatory response. Moreover, the immortalized cells were not available with tumorigenic ability. Therefore, ihESCs would be serviceable as in vitro cell tool to investigate the pathogenesis of endometriosis.


High salt diet decreases longevity in the spontaneously hypertensive rat.
1. Blood pressure, body weight and mortality rate were examined in female spontaneously hypertensive rats (SHR) of the Okamoto strain maintained on a lifelong high (5% NaCl w/w) or low (0.1% NaCl w/w) salt diet. 2. A high salt diet increased mortality and decreased body weight in SHR, but failed to significantly alter blood pressure up to 24 months of age. 3. Hence, a low salt diet does not appear to be a long-term risk factor and may protect the SHR from cardiovascular accidents.


Hepatic SREBP signaling requires SPRING to govern systemic lipid metabolism in mice and humans.
The sterol regulatory element binding proteins (SREBPs) are transcription factors that govern cholesterol and fatty acid metabolism. We recently identified SPRING as a post-transcriptional regulator of SREBP activation. Constitutive or inducible global ablation of Spring in mice is not tolerated, and we therefore develop liver-specific Spring knockout mice (LKO). Transcriptomics and proteomics analysis reveal attenuated SREBP signaling in livers and hepatocytes of LKO mice. Total plasma cholesterol is reduced in male and female LKO mice in both the low-density lipoprotein and high-density lipoprotein fractions, while triglycerides are unaffected. Loss of Spring decreases hepatic cholesterol and triglyceride content due to diminished biosynthesis, which coincides with reduced very-low-density lipoprotein secretion. Accordingly, LKO mice are protected from fructose diet-induced hepatosteatosis. In humans, we find common genetic SPRING variants that associate with circulating high-density lipoprotein cholesterol and ApoA1 levels. This study positions SPRING as a core component of hepatic SREBP signaling and systemic lipid metabolism in mice and humans.


The curious case of an isolated right coronary artery aneurysm complicated by thrombosis and distal embolization.
A 40-year-old male patient with no significant medical history was admitted with an inferior ST-segment elevation myocardial infarction. Primary percutaneous coronary intervention revealed a right coronary artery aneurysm, with no evidence of significant coronary disease. We support the hypothesis of aneurysmal thrombus formation with distal embolization.


Metastatic behavior analyses of tetraspanin TM4SF5-expressing spheres in three-dimensional (3D) cell culture environment.
Cancer metastasis involves diverse cellular functions via bidirectional communications between intracellular and extracellular spaces. To achieve development of the anti-metastatic drugs, one needs to consider the efficacy and mode of action (MOA) of the drug candidates to block the metastatic potentials of cancerous cells. Rather than under two-dimensional environment, investigation of the metastatic potentials under three-dimensional environment would be much pharmaceutically beneficent, since it can mimic the in vivo tumor lesions in cancer patients, leading to allowance of drug candidates analyzed in the 3D culture systems to lower failure rates during the anti-metastatic drug development. Here we have reviewed on the analyses of metastatic potentials of certain cancer models in 3D culture systems surrounded with extracellular matrix proteins, which could be supported by TM4SF5- and/or EMT-mediated actions. We particularly focused the initial events of the cancer metastasis, such as invasive outgrowth and dissemination from the cancer cell masses, spheroids, embedded in the 3D gel culture systems. This review summarizes the significance of tetraspanin TM4SF5 and Snail1 that are related to EMT in the metastatic potentials explored in the 3D gel systems.


Synthesis of (Glu-OMe)2-litorin.
The synthesis of the nonapeptide less than Glu-Glu(OMe)-Trp-Ala-Val-Gly-His-Phe-Met-NH2, corresponding to the formula of the (Glu-OMe)2-litorin, is described. The compound has the same chemical and biological properties of the second bombesin like peptide extracted from the skin of the Australian frog Litoria aurea.


Patient preferences for rheumatoid arthritis treatments: results from the national cross-sectional LERACS study.
To investigate the treatment preferences of patients with rheumatoid arthritis (RA) and determine whether these preferences are related to specific disease characteristics. A national survey was designed to collect demographic, disease, treatment, and preference data on RA patients enrolled in 7 private and university hospital clinics in Lebanon. Associations between patient factors and treatment preferences for RA were analyzed by χ2 or Mann-Whitney U test. A total of 693 patients (83% female; 67% aged 41-70 years) consulting 7 trained rheumatologists completed the survey. Most patients (80%) had established RA >24 months, and approximately one-third (34%) were in remission according to the disease activity score in 28 joints (DAS28). Most (87%) were receiving oral agents (60% oral only). Almost two-thirds of patients (64%) expressed a preference for oral treatments, and more than half (53%) ranked doctor's advice as the most influential factor when choosing treatment. In univariable analysis, health coverage, radiographic damage, disease duration, current therapy, and previous side effects were significantly associated with treatment preference. In multivariable analyses, only radiographic damage and current route of administration were independently associated with preference (both P<0.001), with patients with no radiographic damage and those on oral-only therapy being more likely to prefer oral agents. RA patients expressed a preference for oral rather than subcutaneous/intravenous-administered drugs. Understanding patients' preferences may help to inform policymaker decisions.


Human Epidermal Growth Factor Receptor 2/Human Epidermal Growth Factor Receptor 3 PET Imaging: Challenges and Opportunities.
Human epidermal growth factor receptor 2 (HER2) and HER3 provide actionable targets for both therapy and imaging in breast cancer. Further, clinical trials have shown the prognostic impact of receptor status discordance in breast cancer. Intra- and intertumoral heterogeneity of both HER and hormone receptor expression contributes to inherent errors in tissue sampling, and single biopsies are incapable of identifying discordance in biomarker expression. Numerous PET radiopharmaceuticals have been developed to evaluate (or target for therapy) HER2 and HER3 expression. This review seeks to inform on challenges and opportunities in HER2 and HER3 PET imaging in both clinical and preclinical settings.


Spatial organization of ubiquitin ligase pathways orchestrates neuronal connectivity.
Recent studies have revealed that E3 ubiquitin ligases have essential functions in the establishment of neuronal circuits. Strikingly, a common emerging theme in these studies is that spatial organization of E3 ubiquitin ligases plays a critical role in the control of neuronal morphology and connectivity. E3 ubiquitin ligases localize to the nucleus, centrosome, Golgi apparatus, axon and dendrite cytoskeleton, and synapses in neurons. Localization of ubiquitin ligases within distinct subcellular compartments may facilitate neuronal responses to extrinsic cues and the ubiquitination of local substrates. Here, we review the functions of neuronal E3 ubiquitin ligases at distinct subcellular locales and explore how they regulate neuronal morphology and function in the nervous system.


[Prognostic analysis of allogeneic hematopoietic stem cell transplantation for Philadelphia chromosome-positive acute lymphoblastic leukemia in complete remission in the era of tyrosine kinase inhibitors].
Objective: To study the clinical results and prognostic factors for allo-HSCT of Philadelphia chromosome-positive (Ph(+)) acute lymphoblastic leukemia (ALL) in complete remission (CR) in the era of tyrosine kinase inhibitors (TKI) . Methods: We performed a retrospective analysis of the clinical characteristics of 116 patients with Ph(+)ALL who underwent allo-HSCT while in CR. Results: The study population included 72 men and 44 women. The median patient age was 20 years (4-64 years) . The patients received sibling-identical donor (n=21) , haplo (n=77) , and unrelated donor (n=18) HSCT. The overall survival (OS) rate at 5 years was 73.2% (95% CI 63.8% -80.5% ) . In particular, the 5-year OS can reach 87.5% when the time from diagnosis to transplant is <180 days. The 5-years DFS was 61.4% (95% CI 51.8% -69.7% ) , the 5-year molecular and morphology cumulative relapse incidence was 18.5% (95% CI 12.6% -27.3% ) , and the 5-year TRM was 19.9% (95% CI 13.8% -28.7% ) . A multivariate analysis showed that an age range of 15-39 years (HR=2.730, P=0.044) , time from diagnosis to HSCT ≥ 180 days (HR=4.534, P=0.010) , and Ⅲ-Ⅳgrade aGVHD (HR=7.558, P=0.000) were significantly associated with an inferior overall survival. Limited cGVHD subgroup had better OS (HR=0.300, P=0.034) . Sex, WBC count at diagnosis, type of BCR-ABL fusion genes, somatic gene mutations, CR(1) or >CR(1), MRD negative or positive, conditioning regimen based on TBI or Bu, conditioning intensity, donor source, GVHD prophylactic proposal using cyclosporine or tacrolimus, presence/absence of CMV viremia, and presence/absence of EBV viremia were not significantly different in terms of the OS and DFS. Conclusion: Factors influencing the overall survival of Ph(+) ALL patients who underwent allo-HSCT in CR in the TKI era include age, time form diagnosis to HSCT, and aGVHD severity.


Catalase-positive Eikenella corrodens and Eikenella-like isolates of human and canine origin.
Ten catalase-positive isolates and one catalase-negative isolate that had been assigned to Eikenella corrodens were compared to the nomenclatural type strain regarding selected phenotypic and molecular features and chromosomal deoxyribonucleic acid (DNA) relatedness using the spectrophotometric method. Five catalase-positive human isolates were assigned to the genomic species Eikenella corrodens on the basis of high DNA relatedness levels. Three others, among them strain Chen UB 204, exhibited only moderate degrees of DNA relatedness to the type strain and with each other. Two catalase-positive isolates from dogs were closely interrelated, but yielded only low degrees of DNA binding with Eikenella corrodens and the Eikenella-like human isolates. These findings confirm that the human eikenellas comprise more than one genomic species and that the canine strains represent a distinct taxonomic entity. The differentiation of the strains investigated by conventional phenotypic features, hydrolytic enzyme reactions, and cellular carbohydrate patterns was considered.


Bioequivalence Study of Two Formulations of Mifepristone Tablets in Healthy Chinese Subjects Under Fasting Conditions.
A single-dose, open-label, randomized, two-period crossover-design study was conducted to evaluate the bioequivalence of the reference and test formulations of mifepristone tablets. Each subject was randomized at the beginning to receive a 25-mg tablet of the test or the reference mifepristone under fasting conditions during the first period, then received the alternate formulation during the second period following a 2-week washout period. A validated high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method was used to determine the plasma concentrations of mifepristone and its two metabolites (RU42633 and RU42698). Fifty-two healthy subjects were enrolled in this trial, 50 of whom completed the study. The 90% confidence intervals for the log-transformed Cmax , AUC0-t , and AUC0-∞ fell within the accepted 80%-125% range. Throughout the study period, a total of 58 treatment-emergent adverse events were reported. No serious adverse event was observed. In conclusion, the test and reference mifepristone were bioequivalent and well tolerated under fasting conditions.


Intersectionality and public understandings of health inequity in England: learning from the views and experiences of young adults.
Attempts to reduce health inequities in England frequently prioritise some equity dimensions over others. Intersectionality highlights how different dimensions of inequity interconnect and are underpinned by historic and institutionalised power imbalances. We aimed to explore whether intersectionality could help us shed light on young adults' understanding of health inequities. The study incorporatedqualitative thematic analysis of primary data. Online focus groups with young adults (n = 25) aged 18-30 living in three English regions (Greater London; South Yorkshire/Midlands; North-East England) between July 2020 and March 2021. Online semistructured interviews (n = 2) and text-based communication was conducted for participants unable to attend online groups. Young adults described experiencing discrimination, privilege, and power imbalances driving health inequity and suggested ways to address this. Forms of inequity included cumulative, within group, interacting, and the experience of privilege alongside marginalisation. Young adults described discrimination occurring in settings relevant to social determinants of health and said it adversely affected health and well-being. Intersectionality, with its focus on discrimination and identity, can help public health stakeholders engage with young adults on health equity. An upstream approach to improving health equity should consider multiple and intersecting forms of discrimination along with their cultural and institutional drivers.


Differential efficacy of a chlorine dioxide-containing sanitizer against single species and binary biofilms of a dairy-associated Bacillus cereus and a Pseudomonas fluorescens isolate.
Daily exposure to 100 p.p.m. chlorine dioxide of single species and binary biofilms of dairy-associated Bacillus cereus DL5 and Pseudomonas fluorescens M2, attached to stainless steel surfaces in a laboratory flow system, was studied. Surfaces were sampled daily before and after sanitizer treatment and cells and spores dislodged and enumerated by standard methods. Duplicate surfaces were prepared for confocal scanning laser microscopy (CSLM) and scanning electron microscopy. Higher counts of Ps. fluorescens M2 were obtained in single species biofilms, microcolonies stained green (viable) in CSLM images and were closely packed on attachment surfaces. By contrast, higher counts of B. cereus DL5 were obtained in binary biofilms, microcolonies stained green in CSLM images, but were more spread out. Lower spore counts were obtained for B. cereus DL5 in binary biofilms. The survival of Ps. fluorescens M2 cells after exposure to chlorine dioxide was apparently enhanced by the presence of B. cereus DL5 in binary biofilms. By contrast, B. cereus DL5 showed increased susceptibility to sanitizer treatment in the presence of Ps. fluorescens M2. Co-cultured bacteria in biofilms influence each other with respect to attachment capabilities and sanitizer resistance/susceptibility. Binary biofilms endemic in food-processing industries can survive sanitation regimes and may represent reservoirs of product contamination leading to subsequent spoilage and/or food safety risks.


Influenza-attributable years of life lost in older adults in a subtropical city in China, 2012-2017: A modeling study based on a competing risks approach.
The aim of this study was to estimate influenza-attributable years of life lost (YLL) in older adults in subtropical Hefei, China during the years 2012-2017, based on a competing risks approach. The quasi-Poisson model was fitted to weekly numbers of all-cause deaths by 5-year age groups for older adults ≥60 years of age. The product of the weekly influenza-like illness consultation rate and the proportion of specimens that tested positive for influenza was taken as the measurement of influenza activity, which was incorporated into the model as an exploratory variable. Excess deaths associated with influenza were calculated by subtracting baseline deaths (setting influenza activity to zero) from fitted deaths. Influenza-attributable YLL accounting for competing risks was estimated using restricted mean lifetime survival analysis. The annual influenza-attributable YLL was highest in the 75-79 years age group (565 per 100,000 persons, 95% confidence interval 550-580), followed by the 80-84, 70-74, 85-89, 65-69, and 60-64 years age groups. Influenza A(H3N2) virus was associated with higher YLL than A(H1N1) and B viruses. Influenza-attributable YLL accounted for 1.03-1.53% of total YLL, and the proportion would be overestimated to 2.91-7.34% if the traditional Kaplan-Meier method ignoring competing risks was used. Although influenza-associated mortality increased with age, influenza-attributable YLL was found to be highest in the 75-79 years age group.


Synthesis and characteristics of the enantiomers of the cyclic metabonate of methadone.
2-Ethylidene-1,5-dimethyl-3,3-diphenylpyrrolidine (II), a cyclic metabonate of methadone was synthesized from methadone (I) and methadone-N-oxide (III). M-Chloroperbenzoic acid was used to oxidatively demethylate I while sulphur dioxide reductively demethylated III to give II. The importance of these syntheses lies in the fact that there is no attack on the asymmetric carbon atom in I or III, thus providing direct methods of obtaining the optical isomers of II.


TBC domain family, member 15 is a novel mammalian Rab GTPase-activating protein with substrate preference for Rab7.
Ypt/Rabs are Ras-related GTPases that function as key regulators of intracellular vesicular trafficking. Their slow intrinsic rates of GTP hydrolysis are catalyzed by GTPase-activating proteins (GAPs). Ypt/Rab-GAPs constitute a family of proteins that contain a TBC (Tre-2/Bub2/Cdc16) domain. Only three of the 51 family members predicted in the human genome are confirmed Ypt/Rab-GAPs. Here, we report the identification and characterization of a novel mammalian Ypt/Rab-GAP, TBC domain family, member 15 (TBC1D15). TBC1D15 is ubiquitously expressed and localized predominantly to the cytosol. The TBC domain of TBC1D15 exhibits relatively high homology with that of Gyp7p, a yeast Ypt/Rab-GAP. Furthermore, TBC1D15 stimulates the intrinsic GTPase activity of Rab7, and to a lesser extent Rab11, but is essentially inactive towards Rab4 or Rab6. These data increase the number of mammalian TBC domain family members with demonstrated Rab-GAP activity to four, and suggest that TBC1D15 may be involved in Rab7-mediated late endosomal trafficking.


Microbiology of the genitalia of nulliparous and postpartum Savanna brown goats.
A study of the bacterial flora of the genitalia of nulliparous Savanna Brown does was carried out both before breeding and at different intervals postpartum to investigate the type of microbial organisms that could be present in the uterus, cervix and the vagina respectively. Of 29 pre-breeding vaginal swabs, Staphylococcus sp. was isolated from 20 goats, Streptococcus sp. from 15 goats and Micrococcus sp. from four goats. Mycoplasma agalactiae was isolated from five goats. The postpartum vagina did not show any appreciable change in the microbial flora, except that Escherichia coli was encountered in two cases. The uterus yielded E. coli from the goats slaughtered 2 days postpartum; Micrococcus sp. from goats slaughtered 12 days postpartum; Staph. aureus from goats slaughtered 16 days postpartum and Staph. aureus from goats slaughtered 24 days postpartum. Also in these two cases-a goat slaughtered at two days postpartum (dpp) and a goat slaughtered 16 dpp-E. coli was present in the uterus. Other isolates from the uteri of slaughtered goats were Micrococcus sp. (12 dpp), Staph. aureus and Micrococcus sp. (16 dpp) and Staph. aureus (24 dpp).


Genome sequence of five Zetaproteobacteria metagenome-assembled genomes recovered from hydrothermal vent Longqi, Southwest Indian Ridge.
Southwest Indian Ridge (SWIR) increasingly becomes the hot spot of deep-sea mining and extreme life research. Strategies for future environmental conservation are undoubtedly required which makes investigation of microbial iron metabolisms imperative. Through deep metagenome sequencing, five iron-oxidizing Zetaproteoabacteria metagenome-assembled genomes were recovered from Longqi hydrothermal vent in the SWIR. Phylogenetic analysis revealed that two of the MAGs might represent novel genus in Zetaproteobacteria while other MAGs were related to Mariprofundus. Functional profile suggested that they might be aerobic chemolithoautotrophic species with genes encoding cytochrome c oxidase, iron oxidase cyc2 homologs and carbon fixation CBB pathway. Versatile capabilities of synthesizing diverse amino acids and cofactors were indicated while possession of various metal ion transporters could be vital to heavy metal resistance. Our work has provided more understanding about phylogenetic and functional features of iron-oxidizing Zetaproteobacteria, which might be important to investigate iron biogeochemistry and mineral oxidation in SWIR.


Outcome of normal and dysfunctional labor in different racial groups.
Spontaneous labor in patients of different racial groups has been studied relating progress and outcome to whether labor was dysfunctional as defined by the partogram and action line. Forty-three percent of primigravidas and 17.6 to 25.8% of multigravidas passed the action line and had a lower admission cervical dilatation and a longer observed first stage than those patients whose labor progress remained to the left of the action line. White and black primigravidas whose labor progressed to the right of the action line had lower 1 and 5 minute Apgar scores and delivered heavier babies than those to the left. The cesarean section rates were 1.6% and 1.4% (left) and 7.6% and 18.2% (right) in white and black primigravidas, respectively. The cesarean section rate was significantly higher in black primigravidas irrespective of the relationship to the action line due to the high incidence of the complications of hypertension such as fetal distress and abruptio placentae in those in normal labor as well as those in dysfunctional labor due to cephalopelvic disproportion in those patients whose cervimetric progress went to the right of the action line.


Structural basis for potent neutralization of SARS-CoV-2 and role of antibody affinity maturation.
SARS-CoV-2 is a betacoronavirus virus responsible for the COVID-19 pandemic. Here, we determined the X-ray crystal structure of a potent neutralizing monoclonal antibody, CV30, isolated from a patient infected with SARS-CoV-2, in complex with the receptor binding domain (RBD). The structure reveals CV30's epitope overlaps with the human ACE2 receptor binding site thus providing the structural basis for its neutralization by preventing ACE2 binding.


The psychological gains and losses of religious conversion.
A presentation of case material demonstrating productive identify formation and conflict resolution resulting from religious conversion with at the same time significant repression that so affected personal relatedness as to eventually lead to breakdown of adjustment.


Clinical features of sudden sensorineural hearing loss in diabetic patients.
Clinical studies of sudden sensorineural hearing loss (SSNHL) rarely focus on diabetic patients. We attempted to elucidate the clinical features of SSNHL in diabetic patients and to evaluate the factors influencing hearing deficits. Retrospective. A retrospective review of diabetic patients with SSNHL was conducted at National Taiwan University Hospital from 1984 to 2003. The demographic and clinical characteristics, audiometries, and course of hearing recovery were reviewed. Sixty-seven patients (38 men and 29 women) with a mean age of 60.1 +/-11.9 years were recruited. The mean duration of diabetes was 7. 5 +/- 7.7 years. The mean fasting plasma glucose (FPG), postprandial plasma glucose (PPG), and glycosylated hemoglobin (HbA1C) at admission were 12.4 +/- 5.3 mmol/L, 14.5 +/- 5.4 mmol/L, and 9.9 +/- 2.9 mmol/L, respectively. Profound hearing loss was very common in this series (44.8%). Hearing impairment was also noted in the opposite ear, especially in the high frequencies. Presenting symptoms, the duration of diabetes, the FPG, and the HbA1C had no significant correlations with the severity of hearing loss. After adjusting for sex and age, a high PPG level was significantly associated with opposite-ear hearing deficits in the middle frequencies. In follow-up, the low and middle tone hearing thresholds in the lesion ears improved more than that at high frequencies, but they still did not return to baseline. Hearing consistently improved within 2 months after disease onset, but improvements were rare thereafter. With suitable treatment, optimal glycemic control could be achieved even under high-dose steroid regimens. In diabetic patients with SSNHL, hearing loss in the contralateral ear and the profound type hearing loss in the lesion ear were commonly noted. The age and PPG level had significant correlations to contra-ear hearing loss. The poor prognosis of sudden deafness in diabetes patients may be caused by preexisting microvascular lesions in the inner ear, and the PPG level could be a risk factor indicator for cochlear dysfunction in diabetic patients. High-dose glucocorticoid should not be contraindicant in diabetic patients with SSNHL. Whether a longer duration of treatment will result in more hearing improvement is worthy of further study.