Diversity and distribution of benthic dinoflagellates in Tonga include the potentially harmful genera Gambierdiscus and Fukuyoa.
Benthic dinoflagellates that can cause illness, such as ciguatera poisoning (CP), are prevalent around the Pacific but are poorly described in many locations. This study represents the first ecological assessment of benthic harmful algae species in the Kingdom of Tonga, a country where CP occurs regularly. Surveys were conducted in June 2016 in the Tongatapu island group, and in June 2017 across three island groups: Ha'apai, Vava'u, and Tongatapu. Shallow subtidal coastal habitats were investigated by measuring water quality parameters and conducting quadrat surveys. Microalgae samples were collected using either macrophyte collection or the artificial substrate method. Benthic dinoflagellates (Gambierdiscus and/or Fukuyoa, Ostreopsis, and Prorocentrum) were counted using light microscopy, followed by molecular analyses (real-time PCR in 2016 and high throughput sequencing (metabarcoding) in 2017) to identify Gambierdiscus and Fukuyoa to species level. Six species were detected from the Tongatapu island group in 2016 (G. australes, G. carpenteri, G. honu, G. pacificus, F. paulensis, and F. ruetzleri) using real-time PCR. Using the metabarcoding approach in 2017, a total of eight species (G. australes, G. carpenteri, G. honu, G. pacificus, G. cheloniae, G. lewisii, G. polynesiensis, and F. yasumotoi) were detected. Species were detected in mixed assemblages of up to six species, with G. pacificus and G. carpenteri being the most frequently observed. Ha'apai had the highest diversity with eight species detected, which identifies this area as a Gambierdiscus diversity 'hotspot'. Vava'u and Tongatapu had three and six species found respectively. Gambierdiscus polynesiensis, a described ciguatoxin producer and proposed causative agent of CP was found only in Ha'apai and Vava'u in 2017, but not in Tongatapu in either year. Ostreopsis spp. and Prorocentrum spp. were also frequently observed, with Prorocentrum most abundant at the majority of sites. In 2016, the highest number of Gambierdiscus and/or Fukuyoa cells were observed on seagrass (Halodule uninervis) from Sopu, Tongatapu. In 2017, the highest numbers of Gambierdiscus and/or Fukuyoa from artificial substrate samples were recorded in the Halimeda dominant habitat at Neiafu Tahi, Vava'u, a low energy site. This raised the question of the effect of wave motion or currents on abundance measurements from artificial substrates. Differences in detection were noticed between macrophytes and artificial substrates, with higher numbers of species found on artificial substrates. This study provides a baseline of benthic dinoflagellate distributions and diversity for Tonga that may be used for future studies and the development of monitoring programmes.


Self-Report Study of Predictors of Physician Wellness, Burnout, and Quality of Patient Care.
The goal of this self-report study was to examine the relation of work variables, self-rated health and mental health status, and perceived social support to physician wellness, physician burnout, and quality of patient care. We administered a demographics questionnaire, the Physician Wellness Inventory, the Maslach Burnout Inventory, and the Patient Care Scale to a random sample of full physician members of the American Academy of Family Physicians. We performed regression analyses on self-reported health status, work variables, and social support data as predictor variables and the subscales from the Physician Wellness Inventory, Maslach Burnout Inventory, and Patient Care Scale as the outcome variables. The response rate was 22%. Self-reported mental health status significantly predicted all of the wellness scales, the burnout scales, and the quality of patient care. The ability to manage the workload was the second strongest predictor of multiple scales. More work should be done to explore the factors related to physicians' self-ratings of mental health status and what that means to them. Also, it is important to study whether self-rated mental health status is related to objective patient care quality measures.


Defining the Assessment and Treatment Competencies to Deliver Low-Intensity Cognitive Behavior Therapy: A Multi-Center Validation Study.
Despite the vastly increased dissemination of the low-intensity (LI) version of cognitive behavior therapy (CBT) for the treatment of anxiety and depression, no valid and reliable indices of the LI-CBT clinical competencies currently exist. This research therefore sought to develop and evaluate two measures: the low-intensity assessment competency scale (LIAC) and the low-intensity treatment competency scale (LITC). Inductive and deductive methods were used to construct the competency scales and detailed rating manuals were prepared. Two studies were then completed. The first study used a quantitative, fully-crossed design and the second a multi-center, quantitative longitudinal design. In study one, novice, qualified, and expert LI-CBT practitioners rated an LI-CBT assessment session (using the LIAC) and an LI-CBT treatment session (using the LITC). Study two used the LIAC and LITC across four training sites to analyze the competencies of LI-CBT practitioners over time, across raters, and in relation to the actor/patients' feedback concerning helpfulness, the alliance, and willingness to return. Both the LIAC and LITC were found to be single factor scales with good internal, test-retest reliability and reasonable inter-rater reliability. Both measures were sensitive to measuring change in clinical competence. The LIAC had good concurrent, criterion, discriminant, and predictive validity, while the LITC had good concurrent, criterion, and predictive validity, but limited discriminant validity. A score of 18 accurately delineated a minimum level of competence in LI-CBT assessment and treatment practice, with incompetent practice associated with patient disengagement. These observational ratings scales can contribute to the clinical governance of the burgeoning use of LI-CBT interventions for anxiety and depression in routine services and also in the methods of controlled studies.


High-level expression of alternative oxidase protein sequences enhances the spread of viral vectors in resistant and susceptible plants.
The alternative oxidase (AOX) is the terminal oxidase of the cyanide-resistant alternative respiratory pathway in plants and has been implicated in resistance to viruses. When tobacco mosaic virus (TMV) vectors were used to drive very high levels of expression of either AOX or AOX mutated in its active site (AOX-E), virus spread was enhanced. This was visualized as the induction of larger hypersensitive-response lesions after inoculation onto NN-genotype tobacco than those produced by vectors bearing sequences of comparable length [the green fluorescent protein (gfp) gene sequence or antisense aox] or the 'empty' viral vector. Also, in the highly susceptible host Nicotiana benthamiana, systemic movement of TMV vectors expressing AOX or AOX-E was faster than that of TMV constructs bearing gfp or antisense aox sequences. Notably, in N. benthamiana, TMV.AOX and TMV.AOX-E induced symptoms that were severe and ultimately included cell death, whereas the empty vector, TMV.GFP and the TMV vector expressing antisense aox sequences never induced necrosis. The results show that, if expressed at sufficiently high levels, active and inactive AOX proteins can affect virus spread and symptomology in plants.


Temperature-dependent sex determination: current practices threaten conservation of sea turtles.
Temperature determines the sex of hatchling green turtles (Chelonia mydas) produced from eggs incubated in a beach hatchery under different temperature regimens. Cola and cool nests (less than 28 degrees C) produced almost no females (0 to 10 percent) and warm, thermostable nests (greater than 29.5 degrees C) produced almost all females (95 to 100 percent). A few intersex hatchlings were produced at lower temperatures. Since little concern is given to temperatures at which sea turtle eggs are incubated in artificial hatcheries, present conservation practices may be producing all male, all female, or even intersex hatchlings.


The early impact of the global lockdown on post-secondary students and staff: A global, descriptive study.
The aim of this study was to gain a preliminary, broad-level understanding of how the first lockdown impacted post-secondary students, faculty, and staff worldwide. The data were obtained via a global online cross-sectional questionnaire survey using a mixed-method design and disseminated to university students, faculty, and staff from April to November 2020. The data were categorized in four themes/categories: (1) social life and relationships, (2) access to services, (3) health experiences, and (4) impact on mental health well-being. The survey included 27,804 participants from 121 countries and 6 continents. The majority of participants were from Europe (73.6%), female (59.2%), under 30 years of age (64.0%), living in large urban areas (61.3%), %), and from middle-income families (66.7%). Approximately 28.4% of respondents reported that the lockdown negatively impacted their social life, while 21.2% reported the lockdown had a positive impact. A total of 39.2% reported having issues accessing products or services, including essentials, such as groceries, or medical services. In addition, respondents reported an increase in stress and anxiety levels and a decrease in quality of life during the first 2 weeks of the lockdown. The COVID-19 pandemic and lockdown measures had an evident impact on the lives of post-secondary students, faculty, and staff. Further research is required to inform and improve policies to support these populations at both institutional and national levels.


Behavioral inhibition: linking biology and behavior within a developmental framework.
Behavioral inhibition refers to a temperament or style of reacting that some infants and young children exhibit when confronted with novel situations or unfamiliar adults or peers. Research on behavioral inhibition has examined the link between this set of behaviors to the neural systems involved in the experience and expression of fear. There are strong parallels between the physiology of behaviorally inhibited children and the activation of physiological systems associated with conditioned and unconditioned fear. Research has examined which caregiving behaviors support the frequency of behavioral inhibition across development, and work on the interface of cognitive processes and behavioral inhibition reveal both how certain cognitive processes moderate behavioral inhibition and how this temperament affects the development of cognition. This research has taken place within a context of the possibility that stable behavioral inhibition may be a risk factor for psychopathology, particularly anxiety disorders in older children. The current chapter reviews these areas of research and provides an integrative account of the broad impact of behavioral inhibition research.


[Teaching emergency medicine at the University Medical Center Freiburg: establishment of an integrative concept].
Efforts to improve the quality of undergraduate medical education are commonly hampered by limited human and financial resources. This deficiency may be offset by the development of well structured and innovative teaching concepts, which optimize available assets. The newly conceived modular course "Emergency Medicine" at the University Medical Center Freiburg was conducted for the first time in the winter semester 2006/2007. The core of the course is a 3-day practical training period. It provides the possibility to teach a maximum number of medical students with only four lecturers using patient simulators, interactive case scenarios (simulation software MicroSim), and case scenarios with standardized patients. Evaluation of the course revealed standardized patients to be the best of all teaching methods with an overall average grade of 1.1 (patient simulators 1.2, computer simulation 1.4). Of the students, 88% stated that the practical training encouraged their interest in the speciality emergency medicine. The excellent student evaluation results show that the new course "Emergency Medicine" for medical students constitutes a successful balance between the constraint of resource limitation and the goal of excellent medical education.


Does genetic heterogeneity account for the divergent risk of type 2 diabetes in South Asian and white European populations?
South Asians are up to four times more likely to develop type 2 diabetes than white Europeans. It is postulated that the higher prevalence results from greater genetic risk. To evaluate this hypothesis, we: (1) systematically reviewed the literature for single nucleotide polymorphisms (SNPs) predisposing to type 2 diabetes in South Asians; (2) compared risk estimates, risk alleles and risk allele frequencies of predisposing SNPs between South Asians and white Europeans; and (3) tested the association of novel SNPs discovered from South Asians in white Europeans. MEDLINE, Embase, the Cumulative Index to Nursing and Allied Health Literature (CINAHL) and the Cochrane registry were searched for studies of genetic variants associated with type 2 diabetes in South Asians. Meta-analysis estimates for common and novel bi-allelic SNPs in South Asians were compared with white Europeans from the DIAbetes Genetics Replication And Meta-analysis (DIAGRAM) consortium. The population burden from predisposing SNPs was assessed using a genotype score. Twenty-four SNPs from 21 loci were associated with type 2 diabetes in South Asians after meta-analysis. The majority of SNPs increase odds of the disorder by 15-35% per risk allele. No substantial differences appear to exist in risk estimates between South Asians and white Europeans from SNPs common to both groups, and the population burden also does not differ. Eight of the 24 are novel SNPs discovered from South Asian genome-wide association studies, some of which show nominal associations with type 2 diabetes in white Europeans. Based on current literature there is no strong evidence to indicate that South Asians possess a greater genetic risk of type 2 diabetes than white Europeans.


[Disorders of myocardial contraction and cardiomyocyte ultrastructure after emotional stress].
By means of physiological, electron microscopical and histochemical methods, changes in intensity of mechanisms participating in calcium transport and the cations localization in cardiomyocytes have been studied, the changes resulted from an emotional-painful stress. After the stress, the contractile function of the rat isolated heart becomes more dependent on calcium concentration in the perfusate. As demonstrate combined electron microscopical and histochemical methods and, first of all, determination of calcium antimonate in the cardiomyocytes, together with increased cardiac function, an elevated amount of calcium localized in sarcoplasm of cardiomyocytes is observed, especially in the subsarcolemmic area. The data obtained demonstrate that the membrane mechanisms of calcium transport are injured and calcium content in the sarcoplasm is increased; that can play a definite role in the development of focal lesions in the myocardium at a stress.


Hyperbaric oxygen in chronic cluster headaches: influence on serotonergic pathways.
A controlled study was done with the aim of assessing the efficacy of hyperbaric oxygen (HBO2) in cluster headache and of studying the possible influence of this therapeutic approach on serotonergic pathways. Fourteen patients, aged between 26 and 56 yr, suffering from the chronic form of cluster headache were treated with HBO2 (n = 10) or environmental air (placebo) ( n = 4) during the 15 sessions of exposure (lasting 30 min each) in the hyperbaric chamber. The influence of this procedure on serotonergic pathways of pain was monitored by means of study of serotonin binding to mononuclear cells before and after the treatment for both subgroups. All of the treated 14 chronic cluster headache patients completed the study. In the subgroup treated with the placebo, no particular modifications on the number of attacks and of analgesic consumption as well as no change in the specific binding curve of serotonin to mononuclear cells were observed, whereas in the subgroup treated with HBO2 the clinical effectiveness and the appearance of plateau in the binding curves indicated that the oxygen therapy could act through serotonergic pathways.


Seasonality in Violent and Nonviolent Methods of Suicide Attempts: A Cross-Sectional Study on Systematic Registry Data.
Little attention has been paid to seasonality in suicide in Iran. Time pattern in suicide deaths and suicide attempts for some related factors such as gender, mental disorders has been found. In present study, we focus on suicide methods and the association with seasonality and other putative covariates such as gender. Through a cross-sectional study, overall identified suicide attempts and suicide deaths in the province of Ilam from 1 January 2010 and 31 December 2014 were enrolled. We used Edwards' test for test of seasonality in suicide methods. Seasonal effect (peak/trough seasons) and (deaths/attempts suicide) was explored by ratio statistics, the null hypothesis being that the attempted suicides in each method group are evenly distributed over a year. More suicide attempts by hanging 29.4% and self-immolation 41.4% were observed in spring and differ by season pattern in both genders. The overall distribution of suicides by violent and non-violent methods was (males x2=6.3, P=0.041, females x2=7.7, P=0.021) and (males x2=44.5, P=0.001, females x2=104.7, P=0.001), respectively. The peak and trough seasons was observed in taking medications and self-poisoning for spring and winter. Suicide with alcohol was no differ by season pattern (x2=1.0, P=0.460). Suicide in Ilam illustrates a significant seasonality for both violent and non-violent methods of suicide, in both genders, the two peaks were observed in spring and autumn for violent suicides, and spring and summer in non-violent suicides.


High school teacher ratings of academic, social, and behavioral difficulties: Factor structure and normative data for the School Functioning Scale.
Because academic, behavioral, and social difficulties are highly prevalent among U.S. high school students, teacher report is a critical component of comprehensive screening and assessment of adolescents. Currently available teacher questionnaires have limited utility in high school settings because of time demands and lesser opportunity to observe a wide variety of student behaviors relative to elementary school teachers. The School Functioning Scale (SFS) is a 9-item measure designed to efficiently obtain teacher perceptions of specific indicators of student performance across academic, behavior, and social functioning. Online SFS ratings were collected for 799 9th through 12th grade high school students (50% female) from 400 teachers (63.7% female). Teachers also reported each student's subject grades and percentage of assignments completed within the past month. Exploratory and confirmatory factor analyses indicated a general factor and three-factor structure for the SFS: academic, behavior, and social factors. The general factor demonstrated adequate levels of internal consistency and evidence of convergent validity. Findings regarding the group factors less robust. The factor structure was consistent across age, gender, and ethnicity groups. Normative data along with suggested cut-points for screening and assessment purposes are provided. Findings support the potential use of SFS teacher ratings for multimethod assessment and, possibly, screening and treatment evaluation. (PsycINFO Database Record (c) 2019 APA, all rights reserved).


Isolation and partial amino acid sequence of bacteriocins produced by Lactobacillus acidophilus.
Bacteriocins produced by Lactobacillus acidophilus JCM 1023, JCM 1028, JCM 1021, JCM 1229, and JCM 5342 were active against closely related lactobacilli. These bacteriocins were purified and partial sequenced. Bacteriocin activities of L. acidophilus JCM 1023 and JCM 1028 were associated with two components. On the basis of N-terminal amino acid sequencing and the molecular masses, it is interpreted that these two-component bacteriocins are identical to acidocin J1132, a bacteriocin from L. acidophilus JCM 1132 [Tahara et al., Appl. Environ. Microbiol., 62, 892-897 (1996)]. Other bacteriocins were single-peptide bacteriocins.


Rationale for eliminating Staphylococcus breakpoints for β-lactam agents other than penicillin, oxacillin or cefoxitin, and ceftaroline.
Due to the ongoing concern about the reliability of Staphylococcus breakpoints (interpretive criteria) for other β-lactam agents, the Clinical and Laboratory Standards Institute recently approved the elimination of all breakpoints for antistaphylococcal β-lactams except for penicillin, oxacillin or cefoxitin, and ceftaroline. Routine testing of penicillin and oxacillin or cefoxitin should be used to infer susceptibility for all β-lactams with approved clinical indications for staphylococcal infections. It is critical for laboratories to reject requests for susceptibility testing of other β-lactams against staphylococci and to indicate that susceptibility to these agents can be predicted from the penicillin and oxacillin or cefoxitin results. This article reviews β-lactam resistance mechanisms in staphylococci, current antimicrobial susceptibility testing and reporting recommendations for β-lactams and staphylococci, and microbiologic data and clinical data supporting the elimination of staphylococcal breakpoints for other β-lactam agents.


Isotopic Evidence for Disrupted Copper Metabolism in Amyotrophic Lateral Sclerosis.
Redox-active metals are thought to be implicated in neurodegenerative diseases including amyotrophic lateral sclerosis (ALS). To address this point, we measured the concentrations of 12 elements and, for the first time, the stable isotope compositions of copper (redox-active) and zinc (redox-inactive) in human cerebrospinal fluids of 31 patients with ALS, 11 age-matched controls (CTRL), and 14 patients with Alzheimer disease. We first show that metal concentrations weakly discriminate patients with ALS from the two other groups. We then report that zinc isotopic compositions are similar in the three groups, but that patients with ALS have significantly 65copper-enriched isotopic compositions relative to CTRL and patients with AD. This result unambiguously demonstrates that copper is implicated in ALS. We suggest that this copper isotopic signature may result from abnormal protein aggregation in the brain parenchyma, and propose that isotopic analysis is a potential tool that may help unraveling the molecular mechanisms at work in ALS.


Designed Polyhydroxyproline Helical Peptide with Ultrarobust Antifouling Capability for Electrochemical Sensing in Diverse Complex Biological Fluids.
Developing a generalized strategy for the nonfouling detection of biomarkers in diverse biological fluids presents a significant challenge. Herein, a polyhydroxyproline helical peptide (PHHP) was designed and adopted to fabricate electrochemical microsensors capable of detecting targets in various biological media. The PHHP possessed unique properties such as strong hydrophilicity, rigid structure, and lack of ionizable side-chain groups. Compared with common zwitterionic peptides (ZIPs), the PHHP exhibited similar antifouling capability but exceptional stability, allowing its antifouling performance to be unaffected by environmental alteration. The PHHP can prevent biofouling even in fluctuating pH conditions, high ionic strength environments, and the presence of high-valence ions and resist the protease hydrolysis. The PHHP-modified carbon fiber microelectrode was further immobilized with an aptamer to construct an antifouling microsensor for cortisol detection across diverse biofluids, and the microsensor exhibited acceptable accuracy and higher sensitivity than the ELISA method. In addition, different biological samples of mice were collected in situ using a microsensing device, and cortisol levels were analyzed in each specifically tailored region. This nonfouling sensing strategy based on PHHP allows a comprehensive assessment of biomarkers in both spatial and temporal dimensions in diverse biological environments, holding promising potential for early disease diagnosis and real-time health monitoring.


Diversity in clinical trial inclusion for peripheral artery disease lower extremity endovascular interventions: a systematic review protocol.
Aim: This review provides a study protocol for a systematic review of peripheral artery disease (PAD) clinical trials to examine the eligibility criteria, demographic representation, and enrollment strategies among PAD patients undergoing lower extremity (LE) endovascular interventions. Methods: This systematic review will be conducted according to the Cochrane Collaboration methodology for systematic reviews and following the Preferred Reporting Items for Systematic Reviews and Meta-Analysis Protocols (PRISMA-P). Eligible studies will include randomized controlled trials (RCTs) between January 2012 and December 2022. The primary outcome will be a description and summary of the frequency of the reporting of demographic characteristics. The feasibility of a meta-analysis or meta-regression will be explored, but if determined to be infeasible, the Synthesis Without Meta-analysis (SWiM) reporting guideline will be followed for the reporting of findings. Discussion: The findings may help to quantify existing inequities in clinical trial participation that may be addressed through optimizing enrollment strategies for future PAD trials. Systematic review registration: PROSPERO (CRD42022378304).


Efficacy of Sinopharm® COVID-19 Vaccine in Hemodialysis Patients: A Preliminary Report.
SARS-CoV-2 infection have been reported to have a greater mortality rate in adults receiving dialysis, as compared to general population. Hence, vaccination is very important in this vulnerable population group, in order to achieve an acceptable level of immunity. The aim of this study was to compare the level of anti-SARS-CoV-2 anti-spike protein receptor-binding domain IgG neutralizing antibody before and after vaccination with two doses of Sinopharm® vaccine, in patients undergoing hemodialysis. Ninety patients on maintenance in-center hemodialysis received two doses of Sinopharm® COVID-19 vaccine with an interval of about 28 days. Anti-SARS-CoV-2 anti-spike protein receptor-binding domain IgG (Anti-RBD) neutralizing antibody was measured with an ELISA kit. All statistical analyses were performed by SPSS-26 software. The absolute mean (± SE) change in antibody titer following full-scheduled vaccination was 8.98 ± 1.49 µg/mL. The rate of seroconversion was 31.1% after two doses of vaccine. In addition, the rate of seroconversion was higher in those with a history of COVID-19 than in those without a history of COVID-19. The administration of booster doses, doubling of the dose in each episode of vaccination schedule as well as combination of different vaccine platforms are recommended to increase COVID-19 vaccine efficacy in hemodialysis patients. DOI: 10.52547/ijkd.7024.


Political differences in free will belief are associated with differences in moralization.
In 14 studies, we tested whether political conservatives' stronger free will beliefs were linked to stronger and broader tendencies to moralize and, thus, a greater motivation to assign blame. In Study 1 (meta-analysis of 5 studies, n = 308,499) we show that conservatives have stronger tendencies to moralize than liberals, even for moralization measures containing zero political content (e.g., moral badness ratings of faces and personality traits). In Study 2, we show that conservatives report higher free will belief, and this is statistically mediated by the belief that people should be held morally responsible for their bad behavior (n = 14,707). In Study 3, we show that political conservatism is associated with higher attributions of free will for specific events. Turning to experimental manipulations to test our hypotheses, we show the following: when conservatives and liberals see an action as equally wrong there is no difference in free will attributions (Study 4); when conservatives see an action as less wrong than liberals, they attribute less free will (Study 5); and specific perceptions of wrongness account for the relation between political ideology and free will attributions (Study 6a and 6b). Finally, we show that political conservatives and liberals even differentially attribute free will for the same action depending on who performed it (Studies 7a-d). These results are consistent with our theory that political differences in free will belief are at least partly explicable by conservatives' tendency to moralize, which strengthens motivation to justify blame with stronger belief in free will and personal accountability. (PsycInfo Database Record (c) 2021 APA, all rights reserved).


Metagenomic Signatures of Gut Infections Caused by Different Escherichia coli Pathotypes.
Escherichia coli is a leading contributor to infectious diarrhea and child mortality worldwide, but it remains unknown how alterations in the gut microbiome vary for distinct E. coli pathotype infections and whether these signatures can be used for diagnostic purposes. Further, the majority of enteric diarrheal infections are not diagnosed with respect to their etiological agent(s) due to technical challenges. To address these issues, we devised a novel approach that combined traditional, isolate-based and molecular-biology techniques with metagenomics analysis of stool samples and epidemiological data. Application of this pipeline to children enrolled in a case-control study of diarrhea in Ecuador showed that, in about half of the cases where an E. coli pathotype was detected by culture and PCR, E. coli was likely not the causative agent based on the metagenome-derived low relative abundance, the level of clonality, and/or the virulence gene content. Our results also showed that diffuse adherent E. coli (DAEC), a pathotype that is generally underrepresented in previous studies of diarrhea and thus, thought not to be highly virulent, caused several small-scale diarrheal outbreaks across a rural to urban gradient in Ecuador. DAEC infections were uniquely accompanied by coelution of large amounts of human DNA and conferred significant shifts in the gut microbiome composition relative to controls or infections caused by other E. coli pathotypes. Our study shows that diarrheal infections can be efficiently diagnosed for their etiological agent and categorized based on their effects on the gut microbiome using metagenomic tools, which opens new possibilities for diagnostics and treatment.IMPORTANCEE. coli infectious diarrhea is an important contributor to child mortality worldwide. However, diagnosing and thus treating E. coli infections remain challenging due to technical and other reasons associated with the limitations of the traditional culture-based techniques and the requirement to apply Koch's postulates. In this study, we integrated traditional microbiology techniques with metagenomics and epidemiological data in order to identify cases of diarrhea where E. coli was most likely the causative disease agent and evaluate specific signatures in the disease-state gut microbiome that distinguish between diffuse adherent, enterotoxigenic, and enteropathogenic E. coli pathotypes. Therefore, our methodology and results should be highly relevant for diagnosing and treating diarrheal infections and have important applications in public health.


End-of-life decision-making in Canada: the report by the Royal Society of Canada expert panel on end-of-life decision-making.
This report on end-of-life decision-making in Canada was produced by an international expert panel and commissioned by the Royal Society of Canada. It consists of five chapters. Chapter 1 reviews what is known about end-of-life care and opinions about assisted dying in Canada. Chapter 2 reviews the legal status quo in Canada with regard to various forms of assisted death. Chapter 3 reviews ethical issues pertaining to assisted death. The analysis is grounded in core values central to Canada's constitutional order. Chapter 4 reviews the experiences had in a number of jurisdictions that have decriminalized or recently reviewed assisted dying in some shape or form. Chapter 5 provides recommendations with regard to the provision of palliative care in Canada, as well as recommendations for reform with respect to the various forms of assisted death covered in this document.


Use of Dietary Supplements and Perceived Knowledge among Adults Living with Fibromyalgia in Norway: A Cross-Sectional Study.
Fibromyalgia syndrome (FMS) is a complex medical condition characterized by widespread musculoskeletal pain. To date, no gold standard treatment has been developed, and persons with FMS often seek alternative methods to control their symptoms, such as dietary supplements (DS). This study aimed to describe the use of DS in persons living with FMS and examine the associations between the use of DS and its potential predictors. We recruited a convenience sample of 504 participants (≥18 years) living with FMS. The main outcome variables included estimated expenditure on DS in the last 12 months in Norwegian kroner (NOK) and the differences between the groups of users and non-users of DS. Of the 504 participants, 430 reported having used DS, and the mean amount of money spent in the previous year was determined to be NOK 2300. The most common DS reported were vitamin D, magnesium, and omega-3 fatty acids. The predictors of being a DS user were high education, high self-reported knowledge of DS but low overall knowledge of health claims. Users of DS marketed for muscles/joints appear to spend more money on DS. The increasing availability of DS and aggressive advertising in the media through health claims stipulate the need for interventions that lead to informed decisions about DS.


Effects of material morphology on the phototoxicity of nano-TiO2 to bacteria.
Nanostructured titania (nano-TiO2) is produced in diverse shapes, but it remains largely unknown how tuning the morphology of nano-TiO2 may alter its toxicity. Herein, we show that material morphology plays a critical role in regulating the phototoxicity of nano-TiO2 to bacteria. Low-dimensional nano-TiO2, including nanotubes, nanorods, and nanosheets, were synthesized hydrothermally, and their effects on the bacterial viability of Escherichia coli and Aeromonas hydrophila were compared to spherical nanostructures (anatase nanospheres and P25). Results reveal that TiO2 nanotubes and nanosheets are less phototoxic than their rod- and sphere-shape counterparts under simulated solar irradiation. None of the tested nano-TiO2 shows toxicity in the dark. In contrast to their diminished phototoxicity, however, TiO2 nanotubes and nanosheets exhibit comparable or even higher photoactivity than other nanostructures. Observations by scanning transmission electron microscopy suggest that material morphology influences nano-TiO2 phototoxicity by governing how nano-TiO2 particles align at the bacterial cell surface. Overall, when comparing materials with different morphologies and dimensionality, nano-TiO2 phototoxicity is not a simple function of photocatalytic reactivity or ROS production. Instead, we propose that the evaluation of nano-TiO2 phototoxicity encompasses a three-pronged approach, involving the intrinsic photoactivity, aggregation of nano-TiO2, and the nano-TiO2/bacteria surface interactions.


Balancing division and expansion during maize leaf morphogenesis: analysis of the mutant, warty-1.
Cell division and expansion are growth events that contribute to the developing shape, or morphogenesis, of a plant. Division and expansion are coordinated to the extent that plant organs, such as leaves, generally portray a predictable cellular pattern. To dissect the relationship between division and expansion, and to test for the role of each during morphogenesis, we have identified a recessive mutation warty-1 that produces a primary defect in cell size and shape in mutant leaves. Warty-1 mutant plants are similar to non-mutant siblings in terms of flowering time, overall plant size and leaf shape. Mature adult leaves have raised warts, consisting of excessively enlarged cells, that appear in patchy distribution throughout the blade. Cell wall deposition is abnormal or incomplete, suggesting cytokinesis is also affected, either directly or indirectly. Cells first increase in size at specific positions, which correspond to predictable cell dimensions of a developing 1 cm leaf. Once mutant cells exceed 133% normal size, cytokinesis becomes abnormal. As differentiation progresses, cells that appear normal in the mutant are actually dividing faster and are smaller than comparable cells in non-mutant siblings. These results suggest that (1) cells may compensate for growth defects by altering their cell cycle and that (2) proper execution of cytokinesis may require that cell size ratios are properly maintained.


High-Throughput Quantification of Surface Protein Internalization and Degradation.
Cell surface proteins are key regulators of fundamental cellular processes and, therefore, often at the root of human diseases. Thus, a large number of targeted drugs which are approved or under development act upon cell surface proteins. Although down-regulation of surface proteins by many natural ligands is well-established, the ability of drug candidates to cause internalization or degradation of the target is only recently moving into focus. This property is important both for the pharmacokinetics and pharmacodynamics of the drug but may also constitute a potential resistance mechanism. The enormous numbers of drug candidates targeting cell surface molecules, comprising small molecules, antibodies, or alternative protein scaffolds, necessitate methods for the investigation of internalization and degradation in high throughput. Here, we present a generic high-throughput assay protocol, which allows the simultaneous and independent quantification of internalization and degradation of surface proteins on a single-cell level. Because we fuse a HaloTag to the cell surface protein of interest and exploit the differential cell permeability of two fluorescent HaloTag ligands, no labeling of the molecules to be screened is required. In contrast to previously described approaches, our homogeneous assay is performed with adherent live cells in a 96-well format. Through channel rescaling, we are furthermore able to obtain true relative abundances of surface and internal protein. We demonstrate the applicability of our procedure to three major drug targets, EGFR, HER2, and EpCAM, examining a selection of well-investigated but also novel small molecule ligands and protein affinity reagents.


Sub-wavelength terahertz beam profiling of a THz source via an all-optical knife-edge technique.
Terahertz technologies recently emerged as outstanding candidates for a variety of applications in such sectors as security, biomedical, pharmaceutical, aero spatial, etc. Imaging the terahertz field, however, still remains a challenge, particularly when sub-wavelength resolutions are involved. Here we demonstrate an all-optical technique for the terahertz near-field imaging directly at the source plane. A thin layer (<100 nm-thickness) of photo carriers is induced on the surface of the terahertz generation crystal, which acts as an all-optical, virtual blade for terahertz near-field imaging via a knife-edge technique. Remarkably, and in spite of the fact that the proposed approach does not require any mechanical probe, such as tips or apertures, we are able to demonstrate the imaging of a terahertz source with deeply sub-wavelength features (<30 μm) directly in its emission plane.


Myelofibrosis in a cat (abstract).
A 2-year-old female domestic shorthair cat was presented with ill-health and marked ascites. Clinically, there was marked pallor of mucous membranes. Two haematological examinations and transudate-exudate analyses of peritoneal fluid were conducted 1 week apart, as well as a coagulation screen and routine serum biochemistry. The haematological examinations revealed a severe persistent non-regenerative anaemia. There were no abnormalities within the neutrophil and lymphocyte cell lines. Very occasional nucleated red blood cells (RBCs) in peripheral circulation and some RBC agglutination were seen. Analysis of peritoneal fluid disclosed a modified transudate, and small numbers of erythroid and myeloid precursors were observed on the second occasion. The cat was FIV/FeLV-negative, and there were no coagulation abnormalities. Following necropsy and histopathological examination, myelofibrosis and extramedullary haematopoiesis were seen. Thoracic masses were hyperplastic lymph nodes in which there was evidence of extramedullary haematopoiesis. The exact cause of the myelofibrosis was not obvious but it may have been a case of myeloid metaplasia, however not all the characteristics of this entity were present.


Gene and translation initiation site prediction in metagenomic sequences.
Gene prediction in metagenomic sequences remains a difficult problem. Current sequencing technologies do not achieve sufficient coverage to assemble the individual genomes in a typical sample; consequently, sequencing runs produce a large number of short sequences whose exact origin is unknown. Since these sequences are usually smaller than the average length of a gene, algorithms must make predictions based on very little data. We present MetaProdigal, a metagenomic version of the gene prediction program Prodigal, that can identify genes in short, anonymous coding sequences with a high degree of accuracy. The novel value of the method consists of enhanced translation initiation site identification, ability to identify sequences that use alternate genetic codes and confidence values for each gene call. We compare the results of MetaProdigal with other methods and conclude with a discussion of future improvements. The Prodigal software is freely available under the General Public License from http://code.google.com/p/prodigal/.


Causal relationship between rheumatoid arthritis and hypothyroidism or hyperthyroidism: a bidirectional two-sample univariable and multivariable Mendelian randomization study.
The causal relationship between Rheumatoid arthritis (RA) and hypothyroidism/hyperthyroidism remains controversial due to the limitations of conventional observational research, such as confounding variables and reverse causality. We aimed to examine the potential causal relationship between RA and hypothyroidism/hyperthyroidism using Mendelian randomization (MR). We conducted a bidirectional two-sample univariable analysis to investigate the potential causal relationship between hypothyroidism/hyperthyroidism and RA. Furthermore, we performed a multivariate analysis to account for the impact of body mass index (BMI), smoking quantity, and alcohol intake frequency. The univariable analysis indicated that RA has a causative influence on hypothyroidism (odds ratio [OR]=1.07, 95% confidence interval [CI]=1.01-1.14, P=0.02) and hyperthyroidism (OR=1.32, 95% CI=1.15-1.52, P<0.001). When hypothyroidism/hyperthyroidism was considered as an exposure variable, we only observed a causal relationship between hypothyroidism (OR=1.21, 95% CI=1.05-1.40, P=0.01) and RA, whereas no such connection was found between hyperthyroidism (OR=0.91, 95% CI=0.83-1.01, P=0.07) and RA. In the multivariate MR analyses, after separately and jointly adjusting for the effects of daily smoking quantity, alcohol intake frequency, and BMI, the causal impact of RA on hypothyroidism/hyperthyroidism and hypothyroidism on RA remained robust. However, there is no evidence to suggest a causal effect of hyperthyroidism on the risk of RA (P >0.05). Univariate and multivariate MR analyses have validated the causal association between RA and hypothyroidism/hyperthyroidism. Hypothyroidism confirmed a causal relationship with RA when employed as an exposure variable, whereas no such relationship was found between hyperthyroidism and RA.


A spontaneous low-pathogenic variant of Theiler's virus contains an amino acid substitution within the predominant VP1(233-250) T-cell epitope.
Theiler's murine encephalomyelitis virus (TMEV) induces immune-mediated demyelination after intracerebral inoculation of the virus into susceptible mouse strains. We isolated from a TMEV BeAn 8386 viral stock, a low-pathogenic variant which requires greater than a 10,000-fold increase in viral inoculation for the manifestation of detectable clinical signs. Intracerebral inoculation of this variant virus induced a strong, long-lasting, protective immunity from the demyelinating disease caused by pathogenic TMEV. The levels of antibodies to the whole virus as well as to the major linear epitopes were similar in mice infected with either the variant or wild-type virus. However, persistence of the variant virus in the central nervous system (CNS) of mice was significantly lower than that of the pathogenic virus. In addition, the T-cell response to the predominant VP1 (VP1(233-250)) epitope in mice infected with the variant virus was significantly weaker than that in mice infected with the parent virus, while similar T-cell responses were induced against another predominant epitope (VP2(74-86)). Further analyses indicated that a change of lysine to arginine at position 244 of VP1, which is the only amino acid difference in the P1 region, is responsible for such differential T-cell recognition. Thus, the difference in the T-cell reactivity to this VP1 region as well as the low level of viral persistence in the CNS may account for the low pathogenicity of this spontaneous variant virus.


Spiritual Perspectives on Pain in Advanced Breast Cancer: A Scoping Review.
The aim of the present review was to characterize how pain and spirituality have been conceptualized, assessed, and addressed and how these concepts may be related among women with advanced breast cancer. A scoping review was conducted including publications of various methodologies. Searches were conducted in PubMed, CINAHL, PsycINFO, Cochrane Library, OpenGrey, OAIster, and a large university library database (published 2006-2018). Research questions and criteria were formulated at the outset, followed by identification of publications, charting data, and collating results. Forty-two publications met the inclusion criteria. Most (n = 33) focused exclusively on pain, five pain and spirituality, and four exclusively spirituality. Conceptual definitions were not explicitly provided but were implied. Most assessments used the 0-10 Numeric Rating Scale (pain) and qualitative methods (spirituality). Pain management primarily focused on radiotherapy and pharmaceuticals, and two publications identified spiritual interventions. No publications directly examined the impact of spirituality on pain. Findings of qualitative studies including both concepts suggest the potential value of spirituality as a mechanism to cope with pain. This review identified significant unmanaged pain in women with advanced breast cancer. Women identified dimensions of spirituality as important for coping with their disease. A gap in understanding spirituality and its potential influence on pain in this population was identified.


Cytotoxic T-lymphocyte antigen-4 A49G polymorphism is associated with susceptibility to and severity of alcoholic liver disease in Italian patients.
To determine whether the functional A49G polymorphism of cytotoxic T-lymphocyte antigen-4 (CTLA-4), a T-cell surface molecule that modulates T-lymphocyte activation and influences the risk of developing alcohol-induced autoantibodies, plays a role in susceptibility to alcoholic liver disease (ALD) and influences disease severity in Italian alcohol abusers. One hundred and eighty-three patients with chronic ALD (61 cirrhosis), 115 end-stage HCV cirrhosis, 102 non-alcoholic fatty liver disease (NAFLD), 93 healthy subjects and 43 heavy drinkers without liver disease were studied. CTLA-4 gene polymorphism was analysed by restriction analysis. The frequency of the CTLA-4 polymorphism was higher in patients with ALD than in patients with HCV chronic hepatitis and NAFLD, healthy subjects (P < 0.0001), and heavy drinkers without liver disease (P = 0.02). In patients with ALD, homozygosity for the CTLA-4 polymorphic allele (G/G genotype) was more represented in subjects with cirrhosis (P = 0.047), and independently associated with the risk of cirrhosis (OR 3.5; P = 0.03). The CTLA-4 polymorphic G allele, probably by interfering with the immune response, may confer susceptibility to ALD and, in homozygous state, to alcoholic cirrhosis.


Muscle force-length dynamics during level versus incline locomotion: a comparison of in vivo performance of two guinea fowl ankle extensors.
For a terrestrial animal to move in the complex natural environment, the limb muscles must modulate force and work performance to meet changing mechanical requirements; however, it is not clear whether this is accomplished via a collective shift in function by all limb muscles, or a division of labor among limb muscles. Do muscles differ in their ability to modulate force-length contractile function to meet the mechanical demands of different locomotor tasks? We explore this question by examining the in vivo force-length performance of the guinea fowl Numida meleagris lateral gastrocnemius (LG) and digital flexor-IV (DF-IV), during level and incline locomotion. During level locomotion, the LG and DF-IV exhibit differing muscle fascicle strain patterns: the LG shortens by 10-15% while developing force, whereas the DF-IV undergoes a stretch-shorten cycle with large strain amplitudes and small net strains of 1-8%. Furthermore, the DF-IV operates at higher muscle stresses (92-130 kPa, compared to 23-39 kPa for LG) and possesses a longer tendon, which allows the DF-IV tendon to recover greater elastic energy than the LG tendon. During incline locomotion, these muscles contribute only one-third of the energy expected for their mass, with the DF-IV exhibiting high stride-to-stride variability in work output. While the stretch-shorten cycle of the DF-IV muscle may allow more economic force production, it also leads to large changes in work output with small changes in the relative timing of force and strain. Thus, while the primary determinants of LG work are net strain and mean force, the primary determinant of DF-IV work is the phase relationship between force and strain. Our results suggest that, in addition to influencing a muscle's mechanical performance during steady level locomotion, morphology also affects its capacity and mechanism for altering work output for different locomotor tasks.


Facile synthesis of fluorescent hetero[8]circulene analogues with tunable solubilities and optical properties.
Hetero[8]circulenes are an interesting class of polycyclic heteroaromatic molecules having rigid and planar structures, which are promising in light of their potential applications for OLEDs, OFETs and so forth. Although their synthetic methods have been developed in some specific cases, a facile synthetic protocol of novel hetero[8]circulenes with tunable properties is highly desirable. We herein report the unexpected formation of methoxy-substituted quasi-aza[8]circulene and its conversion into unprecedented triazaoxa[8]circulene. The structures and optical properties were comparatively studied. Remarkably, triazaoxa[8]circulene is highly soluble in THF, acetone and DMSO mainly because of effective hydrogen-bonding of the NH moieties to these solvents. Their highly soluble nature in various solvents enabled us to study the solvent effects of these molecules. In particular, triazaoxa[8]circulene displays a high fluorescence quantum yield of 0.72 in DMSO. Furthermore, enantiomeric separation of highly distorted quasi-aza[8]circulene was successfully achieved by chiral HPLC. Thus, these novel hetero[8]circulene derivatives are practically useful fluorescent nanographene-like molecules with intriguing optical properties.


Risk domains associated with an adolescent alcohol dependence diagnosis.
To determine the contribution of familial, interpersonal, academic and early substance use factors to relative risk for an alcohol dependence (AD) diagnosis in adolescents. Information on 619 adolescents and their 390 sets of biological parents was obtained using the adolescent version of the Child Semi-Structured Assessment for the Genetics of Alcoholism (C-SSAGA) and the adult counterpart of this instrument, the Semi-Structured Assessment for the Genetics of Alcoholism (SSAGA). The C-SSAGA elicits a wide range of environmental, social, and psychiatric diagnostic information. Specific domain scale scores associated with an adolescent AD were computed, and generalized estimating equations (GEE) modeling was used to determine the odds ratio (relative risk) of the specified risk domains for an alcohol dependence diagnosis. Risk factors for a DSM-III-R AD diagnosis included being at least 16 years of age, as well as negative parent-child interactions, school and personal-related difficulties (including the presence of an externalizing or internalizing DSM-III-R non-alcohol-related diagnosis), and early experimentations with a variety of substances. An array of familial, interpersonal, academic and early substance use factors were strongly associated with adolescent AD. Given the findings of this study, further research to determine temporal relationships that might influence the onset of adolescent alcohol dependence is warranted.


Behcet's disease associated with bone marrow failure in Korean patients: clinical characteristics and the association of intestinal ulceration and trisomy 8.
The aim of this study was to determine the clinical characteristics of Behcet's disease (BD) associated with bone marrow failure (BMF), classified as conditions such as myelodysplastic syndrome (MDS) or aplastic anaemia (AA), in Korea. A retrospective analysis was made of 13 patients with BD associated with BMF (MDS 8 cases, AA 5 cases) and 66 patients with BD not associated with BMF. These patients all fulfilled the diagnostic criteria of the international BD study group. BD patients with BMF showed significantly lower leucocyte count, haemoglobin level and platelet count when compared with patients without BMF (P < 0.001). BD patients with BMF had significantly higher serum CRP level at the time of BD diagnosis compared with patients without BMF (P = 0.03). Intestinal lesions were more frequent in BD patients with BMF than those without BMF (61.5% vs 13.6%, P = 0.001). Cytogenetic abnormality was observed in 90.9% of BD patients with BMF. Of the cytogenetic abnormalities, trisomy 8 was most common, occurring in 70% of the patients. In four patients with refractory BD associated with BMF, successful treatment of BMF by haematopoietic stem cell transplantation resulted in clinical remission of BD. Our study indicates that intestinal ulceration is a characteristic finding in BD associated with BMF. It also suggests that cytogenetic aberration, especially trisomy 8, may play an important role in the pathogenesis of BD associated with BMF.


Sulfonylurea receptor -1 (SUR1): genetic and metabolic evidences for a role in the susceptibility to type 2 diabetes mellitus.
The pancreatic B-cell ATP-sensitive potassium channel (K(ATP)) is composed of two distinct subunits, an inwardly rectifying ion channel forming the pore (Kir6.2), and a regulatory subunit, namely the sulfonylurea receptor-1 (SUR1), which binds this widely used class of insulin-secreting drugs. Mutations in the genes encoding Kir6.2 and SUR1 may result in familial persistent hyperinsulinemic hypoglycaemia of infancy, demonstrating their role in the regulation of insulin secretion. Studies in various populations with different ethnic background provided evidence that various alleles of single nucleotide polymorphisms (SNPs) in the SUR1 gene, and to a less extent in the Kir6.2 gene, confer a significantly increased risk for the development of type 2 diabetes mellitus (T2DM). Allelic variations of these SNPs were shown to modulate insulin secretion and insulin sensitivity in vivo, thus providing a pathophysiological background to explain their contribution to the genetic susceptibility to T2DM. The aim of this review is to summarise and discuss the significant results of recent literature on the implication of K(ATP), and particularly of SUR1, in the genetic and pathopysiological mechanisms of T2DM.


Bone Loss in the Posterior Edentulous Mandible with Implant-Supported Overdentures vs Complete Dentures: A Systematic Review and Meta-Analysis.
To analyze the current evidence on bone loss in the posterior edentulous mandible restored with complete dentures (CDs), two-implant-supported overdentures (2-IODs), or four-implant-supported overdentures (4-IODs). A search was conducted in the Ovid MEDLINE, Embase, Web of Science, CINAHL, and Cochrane databases for clinical studies comparing bone loss in posterior edentulous mandibles restored with CDs, 2-IODs, or 4-IODs. A meta-analysis was performed using statistical software to estimate the mean differences in bone loss with 95% CI. The level of significance was set at P < .05. The search strategy identified 2,806 articles, of which 14 met the inclusion criteria. The meta-analysis included 7 two-arm studies comparing CDs vs 2-IODs or 2-IODs vs 4-IODs. No statistically significant difference was found in bone loss between 2-IODs and CDs (mean difference -0.25 [95% CI -0.85 to 0.36]; P = .43), whereas bone loss was significantly lower with 4-IODs than with 2-IODs (mean difference -0.96 [95% CI -1.86 to -0.06]; P = .04). Overall, the data were highly heterogenous (I2 > 74%). 4-IODs can benefit the patient by decreasing bone loss in the posterior edentulous mandible. However, 2-IODs may not be superior to CDs in reducing bone loss in the posterior mandible. A validation of these results is needed through well-designed RCTs.


Association of Aquaporin 7 and 9 with Obesity and Fatty Liver in db/db Mice.
Aquaporin (AQP) 7 and AQP9 are membrane channel proteins called aquaglyceroporins and are related to glucose and lipid metabolism. AQP7 is mainly expressed in white adipose tissue (WAT) and is involved in releasing glycerol into the bloodstream. AQP9 is the glycerol channel in the liver that supplies glycerol to the hepatic cells. In this study, we investigated the relationship between the expression of aquaglyceroporins and lifestyle-related diseases, such as obesity and fatty liver, using 22-week-old db/db mice. Body weight, WAT, and liver weight showed increases in db/db mice. The levels of liver lipids, plasma lipids, insulin, and leptin were also increased in db/db mice. Gene expression related to fatty acid and triglyceride synthesis in the liver was enhanced in db/db mice. In addition, gene and protein expression of gluconeogenesis-related enzymes was increased. Conversely, lipolysis-related gene expression in WAT was reduced. In the db/db mice, AQP9 expression in the liver was raised; however, AQP7 expression in WAT was reduced. These results suggest that in db/db mice, enhanced hepatic AQP9 expression increased the supply of glycerol to the liver and induced fatty liver and hyperglycemia. Additionally, reduced AQP7 expression in WAT is associated with excessive lipid accumulation in adipocytes. Aquaglyceroporins are essential molecules for glucose and lipid metabolism, and may be potential target molecules for the treatment of obesity and lifestyle-related diseases.


Pituitary hormones in the brain: what is their function?
Data concerning the presence in the central nervous system of the anterior and intermediate lobe hormones ACTH, beta-lipotropin, alpha-melanocyte stimulating hormone, beta-endorphin, prolactin, growth hormone, gonadotrophic hormone, and thyrotropin stimulating hormone are reviewed. Available evidence for the ACTH-lipotropin family of peptides indicates that synthesis can occur in brain as well as in pituitary. Although behavioral effects have been described for some of these peptides and their fragments (ACTH, alpha-MSH, beta-endorphin, prolactin), the physiological relevance and the mechanisms of such effects, the nature of the biosynthetic pathways involved, and the factors regulating the brain concentrations of these peptides remain to be explored.


Social and economic barriers to adherence among patients at Livingstone General Hospital in Zambia.
Zambia is one of the countries hardest hit by the human immunodeficiency virus (HIV) and acquired immune deficiency syndrome (AIDS) pandemic with a national HIV prevalence estimated at 14% among those aged 15-49 years in 2012. Antiretroviral therapy (ART) has been available in public health facilities in Zambia since 2003. By early 2016, 65% of the 1.2 million Zambians living with HIV were accessing ART. While access to ART has improved the lives of people living with HIV globally, the lack of adherence to ART is a major challenge to treatment success globally. This article reports on social and economic barriers to ART adherence among HIV patients being attended to at Livingstone General Hospital in Zambia. Livingstone General Hospital is located in the Southern province of Zambia, and had over 7000 patients enrolled for HIV care of whom 3880 patients were on ART. An explorative, qualitative study was conducted with 42 patients on ART where data were collected through six focus group discussions (3 male and 3 female groups) and seven in-depth interviews. Data were audio-recorded and transcribed verbatim and subjected to thematic content analysis. Economic factors such as poverty and unemployment and the lack of food were reported as major barriers to adherence. Furthermore, social factors such as traditional medicine, religion, lack of family and partner support, and disclosure were also reported as critical barriers to adherence to ART. Interventions to improve adherence among ART patients should aim to redress the socio-economic challenges at community and individual levels.


Development and evaluation of a multiplex digital PCR method for sensitive and accurate detection of respiratory pathogens in children.
The emergence of multiplex digital polymerase chain reaction (dPCR) and other detection technologies for respiratory pathogens in recent years has facilitated greater understanding of respiratory virus epidemics. In this study, a multiplex dPCR method was developed and evaluated as a means of detecting five respiratory pathogens in children with acute lower respiratory tract infection (ALRTI). With 139 nasopharyngeal swabs collected from children with ALRTI, pathogens were detected using dPCR and quantitative real-time PCR (qPCR) methods. Of those specimens, dPCR detected 86 positive cases, while qPCR identified 84. Moreover, dPCR exhibited higher sensitivity than qPCR, and displayed no cross-reactivity with common respiratory pathogens. These findings suggest that dPCR-based method could become one of the most promising options for acute respiratory pathogen detection.


Aberrant gene expression in the Arabidopsis SULTR1;2 mutants suggests a possible regulatory role for this sulfate transporter in response to sulfur nutrient status.
Sulfur is required for the biosynthesis of cysteine, methionine and numerous other metabolites, and thus is critical for cellular metabolism and various growth and developmental processes. Plants are able to sense their physiological state with respect to sulfur availability, but the sensor remains to be identified. Here we report the isolation and characterization of two novel allelic mutants of Arabidopsis thaliana, sel1-15 and sel1-16, which show increased expression of a sulfur deficiency-activated gene β-glucosidase 28 (BGLU28). The mutants, which represent two different missense alleles of SULTR1;2, which encodes a high-affinity sulfate transporter, are defective in sulfate transport and as a result have a lower cellular sulfate level. However, when treated with a very high dose of sulfate, sel1-15 and sel1-16 accumulated similar amounts of internal sulfate and its metabolite glutathione (GSH) to wild-type, but showed higher expression of BGLU28 and other sulfur deficiency-activated genes than wild-type. Reduced sensitivity to inhibition of gene expression was also observed in the sel1 mutants when fed with the sulfate metabolites Cys and GSH. In addition, a SULTR1;2 knockout allele also exhibits reduced inhibition in response to sulfate, Cys and GSH, consistent with the phenotype of sel1-15 and sel1-16. Taken together, the genetic evidence suggests that, in addition to its known function as a high-affinity sulfate transporter, SULTR1;2 may have a regulatory role in response to sulfur nutrient status. The possibility that SULTR1;2 may function as a sensor of sulfur status or a component of a sulfur sensory mechanism is discussed.


Structural Basis of the Interaction between Tuberous Sclerosis Complex 1 (TSC1) and Tre2-Bub2-Cdc16 Domain Family Member 7 (TBC1D7).
Mutations in TSC1 or TSC2 cause tuberous sclerosis complex (TSC), an autosomal dominant disorder characterized by the occurrence of benign tumors in various vital organs and tissues. TSC1 and TSC2, the TSC1 and TSC2 gene products, form the TSC protein complex that senses specific cellular growth conditions to control mTORC1 signaling. TBC1D7 is the third subunit of the TSC complex, and helps to stabilize the TSC1-TSC2 complex through its direct interaction with TSC1. Homozygous inactivation of TBC1D7 causes intellectual disability and megaencephaly. Here we report the crystal structure of a TSC1-TBC1D7 complex and biochemical characterization of the TSC1-TBC1D7 interaction. TBC1D7 interacts with the C-terminal region of the predicted coiled-coil domain of TSC1. The TSC1-TBC1D7 interface is largely hydrophobic, involving the α4 helix of TBC1D7. Each TBC1D7 molecule interacts simultaneously with two parallel TSC1 helices from two TSC1 molecules, suggesting that TBC1D7 may stabilize the TSC complex by tethering the C-terminal ends of two TSC1 coiled-coils.


Translational enhancement of mdm2 oncogene expression in human tumor cells containing a stabilized wild-type p53 protein.
The mdm2 oncogene has transforming potential that is activated by overexpression. We previously reported the identification of human choriocarcinoma cell lines that have very high levels of mdm2 proteins as well as elevated levels of a stabilized wild-type p53 protein. Importantly, this mdm2 overexpression resulted from enhanced translation of mdm2 mRNA, a mechanism that had not previously been implicated in mdm2 expression control. The focus of this study was to investigate the breadth of enhanced translation of mdm2 mRNA in human cancers and to elucidate the basis for this translational activation. Here we present evidence that translational enhancement of mdm2 expression occurs in a variety of human tumor cells. Most of these samples also have high levels of wild-type p53 protein. However, there is no evidence for concomitant overexpression of the p53 target genes p21/waf1 and gadd45. Additionally, we demonstrate that the translational enhancement of mdm2 involves a preferential increase in mdm2 transcription that is initiated from the internal p53-responsive promoter region of this gene. The particular mdm2 transcripts that are generated contain a distinct 5' untranslated region and exhibit a significantly enhanced translational efficiency. These data provide a quantitative explanation for the overexpression of mdm2 proteins in this class of human tumors.


A dileucine motif targets E-cadherin to the basolateral cell surface in Madin-Darby canine kidney and LLC-PK1 epithelial cells.
E-cadherin is a major adherens junction protein of epithelial cells, with a central role in cell-cell adhesion and cell polarity. Newly synthesized E-cadherin is targeted to the basolateral cell surface. We analyzed targeting information in the cytoplasmic tail of E-cadherin by utilizing chimeras of E-cadherin fused to the ectodomain of the interleukin-2alpha (IL-2alpha) receptor expressed in Madin-Darby canine kidney and LLC-PK(1) epithelial cells. Chimeras containing the full-length or membrane-proximal half of the E-cadherin cytoplasmic tail were correctly targeted to the basolateral domain. Sequence analysis of the membrane-proximal tail region revealed the presence of a highly conserved dileucine motif, which was analyzed as a putative targeting signal by mutagenesis. Elimination of this motif resulted in the loss of Tac/E-cadherin basolateral localization, pinpointing this dileucine signal as being both necessary and sufficient for basolateral targeting of E-cadherin. Truncation mutants unable to bind beta-catenin were correctly targeted, showing, contrary to current understanding, that beta-catenin is not required for basolateral trafficking. Our results also provide evidence that dileucine-mediated targeting is maintained in LLC-PK(1) cells despite the altered polarity of basolateral proteins with tyrosine-based signals in this cell line. These results provide the first direct insights into how E-cadherin is targeted to the basolateral membrane.


Adsorption of methyl orange from aqueous solution using chitosan/diatomite composite.
A novel chitosan/diatomite composite was prepared by a simple mixture in the mass ratio to remove methyl orange (MO) from aqueous media in this study. The composite adsorbent was characterized by Fourier transform infrared spectroscopy and scanning electron microscopy analysis. The parameters to influence the adsorption of MO were studied under such conditions as kinetics, adsorption isotherm, pH effect, and thermodynamics. The results revealed that adsorption of MO was initially rapid and the equilibrium time was reached after 40 min. The optimal value of the pH was 5.0 for better adsorption. The equilibrium data were well fitted to the Langmuir isotherm compared to the Freundlich isotherm, and exhibited the highest capacity and a removal rate of 88.37% under an initial dye concentration of 50 mg/L. The kinetic data were well described by the pseudo-second order model. The thermodynamic calculations revealed that the sorption was viable, spontaneous, and exothermic under the conditions studied. In addition, the chitosan/diatomite composite had good adsorption and desorption performance with respect to reusability after six cycles. These results showed that the chitosan/diatomite could be considered as a potential adsorbent for the removal of MO in aqueous solution.


[Nonspecific suppressive effect of Ehrlich tumor ascitic fluid on transplantation immunity].
The influence of syngeneic Ehrlich ascites tumour fluid on the survival of allogeneic skin allografts was investigated on CC57BR mice. A 3--4-day delay of the allograft rejection in mice injected with ascitic fluid was revealed. Preliminary transplantation of such allograft to mice with Ehrlich tumour produced no intensification of the immunodepressive action of the ascites fluid obtained from them. The data obtained indicated the preferential significance of the antigen-independent immunosuppressive factors of ascites tumour fluid in the suppression of the transplantation immunity in vivo.


COVID-19 and Immune-Mediated RBC Destruction.
To summarize the epidemiologic, clinical, and laboratory characteristics of autoimmune hemolytic anemia (AIHA) secondary to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection or vaccination. We conducted a systematic review using standardized keyword search to identify all reports of SARS-CoV-2 infection or vaccination and AIHA across PubMed, Web of Science, Scopus, and Google Scholar through September 24, 2021. Fifty patients (mean [SD] age, 50.8 [21.6] years) diagnosed with coronavirus disease 2019 (COVID-19) and AIHA were identified. AIHA subtypes and number of patients were as follows: cold AIHA (n = 18), warm AIHA (n = 14), mixed-type AIHA (n = 3), direct antiglobulin test (DAT)-negative AIHA (n = 1), DAT-negative Evans syndrome (n = 1), Evans syndrome (n = 3), and subtype not reported (n = 10). Mean (SD) hemoglobin at AIHA diagnosis was 6.5 [2.8] g/dL (95% confidence interval, 5.7-7.3 g/dL). Median time from COVID-19 symptom onset to AIHA diagnosis was 7 days. In total, 19% (8/42) of patients with COVID-19-associated AIHA with reported outcomes were deceased. Four patients (mean [SD] age, 73.5 [16.9] years) developed AIHA following SARS-CoV-2 vaccination: Pfizer-BioNTech BNT162b2 vaccine (n = 2); Moderna mRNA-1273 vaccine (n = 1); undisclosed mRNA vaccine (n = 1). AIHA occurred after 1 dose in 3 patients (median, 5 days). SARS-CoV-2 infection and vaccination are associated with multiple AIHA subtypes, beginning approximately 7 days after infectious symptoms and 5 days after vaccination.


The effect of early initiation of antiretroviral treatment in infants on pediatric AIDS mortality in South Africa: a model-based analysis.
Guidelines for treatment of pediatric HIV have recently changed to recommend that all infants who are identified as HIV-infected should start antiretroviral treatment (ART) immediately, regardless of their immunologic or clinical status. This study aims to assess the likely impact of this change in guideline in South Africa. A mathematical model was developed to simulate mother-to-child transmission of HIV, disease progression, and death of HIV-infected children <15 years of age. The model is calibrated to South African data sources, including prevention of mother-to-child transmission program coverage data, pre-ART survival data, ART program statistics, and pediatric HIV prevalence studies. Relative to what would be expected in the absence of early ART initiation, the number of infant AIDS deaths over the 2010-2025 period is expected to drop by 23.6% (95% confidence interval [CI]: 22.5-24.5%) at current levels of polymerase chain reaction (PCR) diagnosis, and by 34.2% (95% CI: 32.7-35.6%) if PCR diagnosis increases to 80% of perinatally infected infants at 2 months. However, the pediatric HIV disease burden has shifted toward older children in recent years. The effect of early ART on total pediatric AIDS mortality during the 2010-2025 period is therefore modest: a 9.8% reduction (95% CI: 7.9-12.6%) at current levels of PCR diagnosis, and a 14.2% reduction (95% CI: 11.4-18.2%) if PCR coverage increases to 80% of perinatally infected infants. The changes in ART guidelines for infants will have a significant impact on pediatric AIDS mortality at young ages, but further efforts are required to reduce the substantial growing AIDS mortality in older children.


Heatwave distortion correction using an improved reference sample compensation method and multispectral digital image correlation.
Heatwave distortions, caused by unevenly distributed temperature and refractive index in the optical path, unavoidably occur in high-temperature digital image correlation (DIC) measurement. To eliminate these distortions, a multispectral DIC-aided reference sample compensation method is proposed. The proposed method first adheres a correcting transparent glass (decorated with fluorescent speckle patterns) onto the test specimen (sprayed with red speckle patterns). Then, by illuminating the specimen with ultraviolet- and red-light sources, the blue light excited from the correcting glass and the red light reflected from the specimen surface can be captured by a 3CCD camera, forming a color image. After separating the recorded color images into red and blue subimages, the original and the correcting displacement fields can be calculated from these two sets of subimages using the subset-based local 2D-DIC algorithm. By point-to-point subtracting the correcting displacement fields from the original ones, the heatwave distortions can be eliminated, and the corrected real displacement fields can be obtained. For validation, static heatwave experiments show the feasibility and accuracy of the proposed method in correcting heatwave distortions. A uniaxial tensile test of an aluminum specimen with a central hole was also performed, further confirming the practicality of the proposed method in correcting heatwave distortions and revealing heatwave-hidden deformation.


[Traumatic arterio-venous fistula. Experience with 9 cases].
Traumatic arteriovenous fistulas (TAF) are rare forms of vascular injuries. The authors report an experience of nine new cases. Three of them result from a minor traumatism, while the six others are iatrogenic. The symptoms and the physical examination are very helpful in the diagnosis. Selective arteriography is the most useful paraclinical examination to confirm the affection and for the choice of the mode of therapy: surgery, selective arterial embolization, alone or used with surgery according to the characteristics and the location of the fistula.


Multidisciplinary treatment approach in Treacher Collins syndrome.
Treacher Collins syndrome (TCS) is a common genetic disorder with high penetrance and phenotypic variability. First and second branchial arches are affected in TCS, resulting in craniofacial and intraoral anomalies such as: severe convex facial profile; mid-face hypoplasia; microtia; eyelid colobomas; mandibular retrognathism; cleft palate; dental hypoplasia; heterotopic teeth; maxillary transverse hypoplasia; anterior open bite; and Angle Class II molar relationship. A high incidence of caries is also a typical finding in TCS patients. Nonetheless, even simple dental restorative procedures can be challenging in this patient population due to other associated medical conditions, such as: congenital heart defects; decreased oropharyngeal airways; hearing loss; and anxiety toward treatment. These patients often require a multidisciplinary treatment approach, including: audiology; speech and language pathology; otorhinolaryngology; general dentistry; orthodontics; oral and maxillofacial surgery; and plastic and reconstructive surgeries to improve facial appearance. This paper's purpose was to present a current understanding of Treacher Collins syndrome etiology, phenotype, and current treatment approaches.


Quantitative flux coupling analysis.
Flux coupling analysis (FCA) aims to describe the functional dependencies among reactions in a metabolic network. Currently studied coupling relations are qualitative in the sense that they identify pairs of reactions for which the activity of one reaction necessitates the activity of the other one, but without giving any numerical bounds relating the possible activity rates. The potential applications of FCA are heavily investigated, however apart from some trivial cases there is no clue of what bottleneck in the metabolic network causes each dependency. In this article, we introduce a quantitative approach to the same flux coupling problem named quantitative flux coupling analysis (QFCA). It generalizes the current concepts as we show that all the qualitative information provided by FCA is readily available in the quantitative flux coupling equations of QFCA, without the need for any additional analysis. Moreover, we design a simple algorithm to efficiently identify these flux coupling equations which scales up to the genome-scale metabolic networks with thousands of reactions and metabolites in an effective way. Furthermore, this framework enables us to quantify the "strength" of the flux coupling relations. We also provide different biologically meaningful interpretations, including one which gives an intuitive certificate of precisely which metabolites in the network enforce each flux coupling relation. Eventually, we conclude by suggesting the probable application of QFCA to the metabolic gap-filling problem, which we only begin to address here and is left for future research to further investigate.


Analysis of corneal injuries in King Mahendra Memorial Eye Hospital Bharatpur, Chitwan.
To determine the corneal injuries pattern. It is a retrospective hospital based analysis of 5504 corneal injuries among the 111781 patients attending King Mahendra Memorial Eye Hospital over a period of 4 years in order to know the lesions pattern for planning purpose. Among the corneal lesions, 59.7% were traumatic and 40.3% were non traumatic lesions, wood stick was the commonest traumatic agent (28.7%), 24.1% cases reported in hospital within 7 days of injury. Corneal ulcer was the most commonest traumatic lesions (48.2%). Clinically 69.2% cases were of bacterial lesions. 71.4% cases underwent medical treatment. 54.24% had good vision after treatment 1,2,3,5. Early treatment with antibiotic drops in a case of corneal injuries restores good vision. Protective glasses while working are the preventive measure against traumatic lesions.


The Hippo pathway regulates apical-domain size independently of its growth-control function.
The Hippo pathway, identified in Drosophila and conserved in vertebrates, regulates tissue growth by promoting cell cycle exit and apoptosis. In addition to their well-characterised overproliferation phenotype, adult Drosophila epithelial cells mutant for the kinases Hippo and Warts have hypertrophic apical domains. Here we examine the molecular basis of this apical hypertrophy and its impact on cell proliferation. In the wing imaginal disc epithelium, we observe increased staining for members of the apical polarity complexes aPKC and Crumbs as well as adherens junction components when Hippo activity is compromised, while basolateral markers are not affected. This increase in apical proteins is correlated with a hypertrophy of the apical domain and adherens junctions. The cell surface localisation of the Notch receptor is also increased in mutant clones, opening the possibility that aberrant receptor signalling may participate in overgrowth of hpo-deficient tissue. Interestingly, however, although the polarity determinant Crumbs is required for the accumulation of apical proteins, this does not appear to significantly contribute to the overproliferation defect elicited by loss of Hippo signalling. Therefore, Hippo signalling controls growth and apical domain size by distinct mechanisms.


Case of the weeping wound: formation of a lacrimal pseudocyst following lateral canthal surgery.
An 87-year-old woman presented with a subcutaneous fluid collection overlying the right lateral orbital rim 2 days after undergoing a lateral tarsal strip. Drainage and excision of fibrinous granulation tissue occurred twice within the first postoperative month, with the formation of a lacrimal pseudocyst. The pseudocyst repeatedly enlarged following drainage, necessitating 4 needle aspirations. A third attempt at excision, coupled with cautery to the wound base, and loose wound closure succeeded in breaking the cycle of pseudocyst recurrence. However, the wound continued to drain, so aggressive cautery and botulinum toxin were used to obliterate the area of presumed lacrimal gland laceration and slow tear production. The possibility of damage to the lacrimal system should be entertained in the scenario of a postoperative weeping wound. Formation of a lacrimal pseudocyst should be considered when a cystic lesion appears following surgical intervention in the lateral canthal region.


Results of a new classification algorithm combining K nearest neighbors and recursive partitioning.
We present results of a new computational learning algorithm combining favorable elements of two well-known techniques: K nearest neighbors and recursive partitioning. Like K nearest neighbors, the method provides an independent prediction for each test sample under consideration, while like recursive partitioning, it incorporates an automatic selection of important input variables for model construction. The new method is applied to the problem of correctly classifying a set of chemical data samples designated as being either active or inactive in a biological screen. Training is performed at varying levels of intrinsic model complexity, and classification performance is compared to that of both K nearest neighbor and recursive partitioning models trained using the identical protocol. We find that the cross-validated performance of the new method outperforms both of these standard techniques over a considerable range of user parameters. We discuss advantages and drawbacks of the new method, with particular emphasis on its parameter robustness, required training time, and performance with respect to chemical structural class.


Evaluating the risk of pathogen transmission from wild animals to domestic pigs in Australia.
Wild animals contribute to endemic infection in livestock as well as the introduction, reintroduction and maintenance of pathogens. The source of introduction of endemic diseases to a piggery is often unknown and the extent of wildlife contribution to such local spread is largely unexplored. The aim of the current study was to quantitatively assess the probability of domestic pigs being exposed to different pathogens from wild animals commonly found around commercial piggeries in Australia. Specifically, this study aims to quantify the probability of exposure to the pathogens Escherichia coli, Salmonella spp. and Campylobacter spp. from European starlings (Sturnus vulgarus); Brachyspira hyodysenteriae, Lawsonia intracellularis and Salmonella spp. from rats (Rattus rattus and Rattus norvegicus); and Mycoplasma hyopneumoniae, Leptospira spp., Brucella suis and L. intracellularis from feral pigs (Sus scrofa). Exposure assessments, using scenario trees and Monte Carlo stochastic simulation modelling, were conducted to identify potential pathways of introduction and calculate the probabilities of these pathways occurring. Input parameters were estimated from a national postal survey of commercial pork producers and from disease detection studies conducted for European starlings, rats and feral pigs in close proximity to commercial piggeries in Australia. Based on the results of the exposure assessments, rats presented the highest probability of exposure of pathogens to domestic pigs at any point in time, and L. intracellularis (median 0.13, 5% and 95%, 0.05-0.23) and B. hyodysenteriae (median 0.10, 0.05-0.19) were the most likely pathogens to be transmitted. Regarding European starlings, the median probability of exposure of domestic pigs to pathogenic E. coli at any point in time was estimated to be 0.03 (0.02-0.04). The highest probability of domestic pig exposure to feral pig pathogens at any point in time was found to be for M. hyopneumoniae (median 0.013, 0.007-0.022) and L. intracellularis (median 0.006, 0.003-0.011) for pigs in free-range piggeries. The sensitivity analysis indicates that the presence and number of wild animals around piggeries, their access to piggeries and pig food and water, and, in the case of feral pigs, their proximity to piggeries, are the most influential parameters on the probability of exposure. Findings from this study support identification of mitigation strategies that could be implemented at on-farm and industry level to minimize the exposure risk from European starlings, rats and feral pigs.


Cardiac Implantable Electronic Devices: A Window Into the Evolution of Conduction Disease in Cardiac Amyloidosis.
This study characterized the relationship between conduction disease and cardiac amyloidosis (CA) through longitudinal analysis of cardiac implantable electronic device (CIED) data. Bradyarrhythmias and tachyarrhythmias are commonly reported in CA and may precede a CA diagnosis, although the natural history of conduction disease in CA is not well-described. Patients with CA (transthyretin amyloidosis cardiomyopathy [ATTR-CM] and light-chain amyloidosis [AL-CA]) and a CIED were identified within the Duke University Health System. Patient characteristics at the time of implantation, including demographics and data relevant to CA diagnosis, cardiac imaging, and CIED were recorded. CIED interrogations were analyzed for pacing and atrial fibrillation (AF) burden, activity level, lead parameters, and ventricular arrhythmia incidence and/or therapy. Thirty-four patients with CA (7 with AL-CA, 27 with ATTR-CM [78% with wild-type]; 82% men) with median age of 75 years and a mean ejection fraction of 42 ± 13% had a CIED implanted for bradycardia (65%) or prevention of sudden cardiac death (35%). CIED implantation preceded CA diagnosis in 14 patients (41%). Over a mean follow-up of 3.1 ± 4.0 years, right ventricular sensing amplitudes decreased but did not result in device malfunction; lead impedances and capture thresholds remained stable. Between post-implantation years 1 and 5, mean ventricular pacing increased from 56 ± 9% to 96 ± 1% (p = 0.003) and AF burden increased from 2 ± 1.3 to 17 ± 3 h/day (p = 0.0002). Ventricular arrhythmias were common (mean episodes per patient per year: 6.7 ± 2.3 [ATTR-CM] and 5.1 ± 3.2 [AL-CA]) but predominately nonsustained; only 1 patient with AL-CA required implantable cardioverter-defibrillator therapy. Longitudinal analysis of CIED data in patients with CA revealed progressive conduction disease, with high AF burden and eventual dependence on ventricular pacing, although lead parameters remained stable. Ventricular arrhythmias were common but predominantly nonsustained, particularly in ATTR-CM.


Gene expression profiling of Escherichia coli in response to interactions with the lettuce rhizosphere.
The objective of this study was to examine transcriptional changes in Escherichia coli when the bacterium was growing in the lettuce rhizoshpere. A combination of microarray analyses, colonization assays and confocal microscopy was used to gain a more complete understanding of bacterial genes involved in the colonization and growth of E. coli K12 in the lettuce root rhizosphere using a novel hydroponic assay system. After 3 days of interaction with lettuce roots, E. coli genes involved in protein synthesis, stress responses and attachment were up-regulated. Mutants in curli production (crl, csgA) and flagella synthesis (fliN) had a reduced capacity to attach to roots as determined by bacterial counts and by confocal laser scanning microscopy. This study indicates that E. coli K12 has the capability to colonize lettuce roots by using attachment genes and can readily adapt to the rhizosphere of lettuce plants. Results of this study show curli production and biofilm modulation genes are important for rhizosphere colonization and may provide useful targets to disrupt this process. Further studies using pathogenic strains will provide additional information about lettuce-E. coli interactions.


Enhancing diagnosis of T-cell lymphoma using non-recombined T-cell receptor sequences.
Clonality assessment, which can detect neoplastic T cells by identifying the uniquely recombined T-cell receptor (TCR) genes, provides important support in the diagnosis of T-cell lymphoma (TCL). BIOMED-2 is the gold standard clonality assay and has proven to be effective in European TCL patients. However, we failed to prove its sensitivity in Taiwanese TCL patients, especially based on the TCRβ gene. To explore potential impact of genetic background in the BIOMED-2 test, we analyzed TCRβ sequences of 21 healthy individuals and two TCL patients. This analysis suggests that genetic variations in the BIOMED-2 primer sites could not explain the difference in sensitivity. The BIOMED-2 test results of the two TCL patients were positive and negative, respectively. Interestingly, a higher percentage (>81%) of non-recombined TCRβ sequences was observed in the test-negative patient than those of the test-positive patient and all healthy individuals (13~66%). The result suggests a new TCR target for enhancing TCL diagnosis. To further explore the hypothesis, we proposed a cost-effective digital PCR assay that quantifies the relative abundance of non-recombined TCRβ sequences containing a J2-2P~J2-3 segment. With the digital PCR assay, bone marrow specimens from TCL patients (n=9) showed a positive outcome (i.e., the relative abundance of the J2-2P~J2-3 sequences ≧5%), whereas non-TCL patients (n=6) gave a negative result. As five of nine TCL patients had a negative BIOMED-2 test result, the J2-2P~J2-3 sequences may improve TCL detection. This is the first report showing the capability of characterizing non-recombined TCR sequences as a supplementary strategy for the BIOMED-2 clonality test.


KIT protein expression and analysis of c-kit gene mutation in adenoid cystic carcinoma.
The c-kit proto-oncogene encodes a transmembrane receptor tyrosine kinase (KIT), which is expressed in several normal human tissues, especially mast cells and interstitial cells of Cajal. Expression of KIT has been noted in several types of neoplasms and gene mutation has been shown as a mechanism of c-kit oncogene activation in some tumors. Recently, a single adnexal adenoid cystic carcinoma (ACC) was reported to demonstrate KIT expression, however, examination of KIT expression or c-kit mutation in ACC of salivary glands has not been performed. We examined archival tissue samples from 30 ACC of major and minor salivary glands for KIT protein expression by immunohistochemistry with a polyclonal antibody and c-kit gene mutation by polymerase chain reaction amplification and DNA sequencing. KIT protein expression was noted in 90% of ACCs. An association between the presence of at least 50% KIT positive neoplastic cells and Grade 3 ACC or a solid growth pattern was observed (P < .05). KIT expression in normal or nonneoplastic salivary gland tissue was absent. No c-kit juxtamembrane domain (exon 11) or phosphotransferase domain (exon 17) mutations were found in any of the tumors examined. In conclusion, KIT protein expression is correlated with tumor grade of salivary ACC. However, gene mutation of exon 11 or exon 17 is not a mechanism of c-kit activation in these neoplasms.


Safety Assessment of Citrus Plant- and Seed-Derived Ingredients as Used in Cosmetics.
The Expert Panel for Cosmetic Ingredient Safety (Panel) assessed the safety of 30 Citrus plant- and seed-derived ingredients, which are most frequently reported to function in cosmetics as fragrances and/or skin conditioning agents. Because final product formulations may contain multiple botanicals, each containing similar constituents of concern, formulators are advised to be aware of these constituents and to avoid reaching levels that may be hazardous to consumers. With Citrus plant- and seed-derived ingredients, the Panel was concerned about the presence of the hydroperoxides of limonene and linalool in cosmetics. Industry should use good manufacturing practices to limit impurities. The Panel reviewed the available data presented and concluded that 18 of these ingredients are safe in the present practices of use and concentration when formulated to be non-irritating and non-sensitizing. The data for the remaining 12 ingredients are insufficient to determine safety.


Modulation of the Aggregation of the Prion-like Protein RepA-WH1 by Chaperones in a Cell-Free Expression System and in Cytomimetic Lipid Vesicles.
The accumulation of aggregated forms of proteins as toxic species is associated with fatal diseases such as amyloid proteinopathies. With the purpose of deconstructing the molecular mechanisms of these type of diseases through a Synthetic Biology approach, we are working with a model bacterial prion-like protein, RepA-WH1, expressed in a cell-free system. Our findings show that the Hsp70 chaperone from Escherichia coli, together with its Hsp40 and nucleotide exchange factor cochaperones, modulates the aggregation of the prion-like protein in the cell-free system. Moreover, we observe the same effect by reconstructing the aggregation process inside lipid vesicles. Chaperones reduce the number of aggregates formed, matching previous findings in vivo. We expect that the in vitro approach reported here will help to achieve better understanding and control of amyloid proteinopathies.


Supraclavicular artery in Japanese: an anatomical basis for the flap using a pedicle containing a cervical, non-perforating cutaneous branch of the superficial cervical artery.
We investigated the morphology of the non-perforating cervical cutaneous branch in the lateral cervical triangle using 65 (130 sides) donated cadavers. We found the branch in 104 of the entire 130 sides (80.0%). In the majority (72.1%), the cutaneous branch did not cross the clavicle or acromion but supplied the dorsolateral cervical area. The branch was originated from the superficial cervical artery close to the posterior belly of the omohyoideus muscle and immediately lateral to the external jugular vein. The comitant vein, if present, drained into the external jugular vein. Our observations and measurements (length and diameter) suggested that the non-perforating cutaneous branch is useful for a pedicle of the dorsolateral cervical flap in Japanese people. However, detailed morphologies differ from the previous studies published in western countries.