Variation in the Volatile Profiles of Black and Manchurian Ash in Relation to Emerald Ash Borer Oviposition Preferences.
Emerald ash borer (EAB; Agrilus planipennis) is a devastating pest of ash (Fraxinus spp.) in its invaded range in North America. Its coevolved Asian hosts are more resistant and less preferred for oviposition than susceptible North American species. We compared EAB oviposition preferences and bark and canopy volatile organic compound (VOC) emissions of resistant Manchurian ash and susceptible black ash, and examined relationships between VOC profiles and oviposition. In the field, black ash was highly preferred for oviposition while no eggs were laid on Manchurian ash, and we found clear differences in the VOC profiles of Manchurian and black ash. We detected 78 compounds emitted from these species, including 16 compounds that elicited EAB antennal activity in prior studies. Four compounds were unique to black and 11 to Manchurian ash. Emission rates of 14 canopy and 19 bark volatiles varied among the two species, including four previously reported as antennally active. Specifically, 7-epi-sesquithujene (bark) emissions were greater from black ash, while β-caryophyllene (canopy), linalool (bark), and α-cubebene (bark) were emitted at higher rates by Manchurian ash. No relationships were found between the emission rate of any single compound or group of compounds (e.g. monoterpenes) suggesting that preference may be based on complex profile combinations. This is the first study to directly compare VOCs of black and Manchurian ash as well as the first to examine bark- and canopy-specific VOCs. The unique bark and canopy VOC profiles of these two species implicates potentially important variation in VOCs between a closely related resistant and susceptible species that provides a foundation for future studies of host preferences of EAB.


Depression of tuberculin reaction in mild and moderate protein-calorie malnourished children following BCG vaccination.
Following BCG vaccination, mild to moderate protein-calorie malnourished children of the marasmic type were tuberculin tested with two types of tuberculin. The indurations of these malnourished children were compared to those of a similarly treated cohort of well-nourished controls. A large group of children who were initially malnourished became well-nourished over the time interval between BCG vaccination and tuberculin testing and these children were compared to the other two cohorts. A significant difference was found between the indurations of the malnourished and the well-nourished. The group which changed nutritional status had an average induration between that of the other two groups. Feeding for two days with a high protein-calorie mixture did not significantly increase the indurations of the malnourished. Supplementation with vitamin A had no discernible effect.


Audit on the use of the barium enema.
All 1358 referrals for barium enemas at Bristol Royal Infirmary in 1981 were studied. The overall diagnostic yield for colonic or distal ileal pathology was 33% with 75 (5.5%) cancers detected. General practitioner requested enemas constituted 25.8% of the total undertaken and their diagnostic yield was equal to hospital outpatient requested enemas. Medical and surgical diagnostic yields were similar for both inpatient and outpatients though the indications varied. Women predominated by 3:2 in all age groups. In the under 40's the yield was low (19.8%) except for inflammatory bowel disease. When related to symptoms the lowest yield was obtained for the investigation of abdominal pain (25%), particularly in women, except in those admitted as inpatients. The clinicians were wrong in diagnosing abdominal or pelvic masses as being of colonic origin in over 50% of cases. It is suggested that yield could be improved by being more selective in patients under 40, more use being made of suitable faecal occult blood testing, and ultrasonic scanning of abdominal masses before barium enema.


Rate of tooth movement and dentoskeletal effects of rapid canine retraction by dentoalveolar distraction osteogenesis: A prospective study.
The purpose of this study was to test the null hypothesis that duration of orthodontic treatment can be significantly reduced by accelerating canine retraction using dentoalveolar distraction (DAD). Thirty-six maxillary canines of 19 patients comprised the DAD group, and 28 canines of 14 patients were included in the distalization group (DG). The initial mean ages were 15.8 ± 1.96 years for the DAD group and 16.02 ± 2.8 years for the DG. A custom-made, rigid, tooth-borne intraoral distraction device was used for the DAD group, and intraoral elastics were applied for canine distalization in the DG. Six skeletal and 11 dental variables were measured for the cephalometric evaluation. Canine retraction was 7.9 ± 1.49 mm in 11.8 ± 1.3 days and canine distal tipping was 11.48° ± 4.37° after DAD; the canines were distalized 5.29 ± 2.01 mm and tipped 13.64° ± 9.54° in 200 ± 57 days in the DG. The rates of posterior canine movement were 0.67 ± 0.14 mm per day after DAD and 0.03 ± 0.01 mm per day in the DG. No significant first molar anchorage loss was observed after DAD, although the DG showed some vertical and sagittal first molar movement. We failed to reject the null hypothesis. DAD can reduce the duration of orthodontic treatment time by accelerating canine retraction in extraction patients without undesirable side effects.


Increase in veterinary prescriptions of azithromycin during the COVID-19 pandemic in Brazil.
With the advent of the pandemic in 2020 and the lack of perspectives on the treatment of COVID-19, numerous therapeutic proposals have emerged, including hydroxychloroquine and azithromycin. Therefore, some studies have shown that in many countries, the demand for azithromycin has increased during the pandemic. In Brazil, antibiotics can only be purchased with a medical, dental, or veterinary prescription. This study aimed to determine whether the number of prescriptions made by veterinarians (which could be used by humans) has increased during the pandemic. Data on the purchase of antibiotics made under veterinary prescriptions in Brazilian pharmacies between 2014 and 2021 were collected. To assess the changes in monthly trends in the use of the selected antibiotics, we applied the Joinpoint regression. The most prescribed antibiotic in all years was cephalexin (35%), followed by amoxicillin (24%). During the pandemic, sales of azithromycin substantially increased. Regression analysis showed that since 2014, azithromycin prescriptions grew by an average of 0.67% per month. At the beginning of the pandemic, the monthly growth rate became 12.64%. When comparing azithromycin sales during the pandemic with the historical average (2014-2019), the increase was 41%. During the pandemic, there was no animal health situation in Brazil that required the use of this antibiotic. Veterinary prescriptions may have been an instrument for human access to azithromycin for the treatment of COVID-19. Stricter enforcement policies are needed to address this problem to avoid antimicrobial resistance.


Advancing health equity through social care interventions.
To use evidence on addressing racism in social care intervention research to create a framework for advancing health equity for all populations with marginalized social identities (e.g., race, gender, and sexual orientation). Such groups have disproportionate social needs (e.g., food insecurity) and negative social determinants of health (SDOH; e.g., poverty). We recommend how the Agency for Healthcare Research and Quality (AHRQ) could advance health equity for marginalized populations through social care research and care delivery. This commentary is informed by a literature review of social care interventions that were affiliated with healthcare systems; input from health equity researchers, policymakers, and community leaders attending the AHRQ Health Equity Summit; and consensus of the authors. We recommend that AHRQ: (1) create an ecosystem that values research on SDOH and the effectiveness and implementation of social care interventions in the healthcare sector; (2) work with other federal agencies to (a) develop position statements with actionable recommendations about racism and other systems that perpetuate marginalization based on social identity and (b) develop aligned, complementary approaches to research and care delivery that address social marginalization; (3) advance both inclusive care delivery and inclusive research teams; (4) advance understanding of racism as a social determinant of health and effective strategies to mitigate its adverse impact on health; (5) advance the creation and scaling of effective strategies for addressing SDOH in healthcare systems, particularly in co-creation with community partners; and (6) require social care intervention researchers to use methods that advance our understanding of social health equity. AHRQ, as a federal agency, could help advance health equity using a range of strategies, including using the agency's levers to ensure AHRQ stakeholders examine and address the unique experiences of socially marginalized populations in SDOH and social care intervention research.


Resistance of Pseudomonas aeruginosa PAO to nalidixic acid and low levels of beta-lactam antibiotics: mapping of chromosomal genes.
Resistance to high concentrations of nalidixic acid in Pseudomonas aeruginosa PAO was due to mutations in one locus designated nalA, which was mapped by transduction between hex-9001 and leu-10. The nalA mutants were cross-resistant to pipemidic acid, a nalidixic acid analog, at relatively low concentrations. Replicative DNA synthesis was resistant to both drugs in permeabilized cells of nalA mutants. A locus coding for low-level resistance to nalidixic acid, nalB, was cotransducible with pyrB, proC, and met-28. The nalB mutants were also resistant to low levels of pipemidic acid, novobiocin, and beta-lactam antibiotics (e.g., carbenicillin, azlocillin, and cefsulodin), but not to other drugs, such as gentamicin, rifampin, kanamycin, or tetracycline. In nalB mutants, DNA replication showed wild-type sensitivity to nalidixic acid, whereas carbenicillin-induced filamentation required higher drug levels than in the wild-type strain. Thus, nalB mutations appear to decrease cell permeability to some antibiotics. The sensitivity of replicative DNA synthesis to nalidixic acid and novobiocin was very similar in P. aeruginosa and Escherichia coli; by contrast, the concentrations of these drugs needed to inhibit growth of P. aeruginosa were higher than those reported for E. coli by one or two orders of magnitude.


Modeling the dynamics of mouse iron body distribution: hepcidin is necessary but not sufficient.
Iron is an essential element of most living organisms but is a dangerous substance when poorly liganded in solution. The hormone hepcidin regulates the export of iron from tissues to the plasma contributing to iron homeostasis and also restricting its availability to infectious agents. Disruption of iron regulation in mammals leads to disorders such as anemia and hemochromatosis, and contributes to the etiology of several other diseases such as cancer and neurodegenerative diseases. Here we test the hypothesis that hepcidin alone is able to regulate iron distribution in different dietary regimes in the mouse using a computational model of iron distribution calibrated with radioiron tracer data. A model was developed and calibrated to the data from adequate iron diet, which was able to simulate the iron distribution under a low iron diet. However simulation of high iron diet shows considerable deviations from the experimental data. Namely the model predicts more iron in red blood cells and less iron in the liver than what was observed in experiments. These results suggest that hepcidin alone is not sufficient to regulate iron homeostasis in high iron conditions and that other factors are important. The model was able to simulate anemia when hepcidin was increased but was unable to simulate hemochromatosis when hepcidin was suppressed, suggesting that in high iron conditions additional regulatory interactions are important.


Targeting of endoplasmic reticulum (ER) stress in gliomas.
Gliomas remain a group of malignant brain tumors with dismal prognosis and limited treatment options with molecular mechanisms being constantly investigated. The past decade, extracellular stress and intracellular DNA damage have been shown to disturb proteostasis leading to Endoplasmic Reticulum (ER) stress that is implicated in the regulation of gene expression and the pathogenesis of several tumor types, including gliomas. Upon ER stress induction, neoplastic cells activate the adaptive mechanism of unfolded protein response (UPR), an integrated signaling system that either restores ER homeostasis or induces cell apoptosis. Recently, the manipulation of the UPR has emerged as a new therapeutic target in glioma treatment. General UPR activators or selective GRP78, ATF6 and PERK inducers have been detected to modulate cell proliferation and induce apoptosis of glioma cells. At the same time, target-specific UPR inhibitors and small molecule proteostasis disruptors, work in reverse to increase misfolded proteins and cause a dysregulation in protein maturation and sorting, thus preventing the growth of neoplastic cells. Herein, we discuss the pathogenic implication of ER stress in gliomas onset and progression, providing an update on the current UPR modifying agents that can be potentially used in glioma treatment.


Spinal hemangioblastoma, syrinx, and hydrocephalus in a two-year-old child.
A two-year-old child presented with an acute inability to bear weight. Radiological investigation revealed a large cervicothoracolumbar syrinx of no known cause. During investigation, acute communicating hydrocephalus developed, which required a shunt. At surgery, a small thoracic spinal cord hemangioblastoma was discovered and excised. Complete recovery with collapse of the syrinx followed. The clinical features of this rare childhood tumor and its associated effects are discussed.


[The experience with the topical application of non-steroidal anti-inflammatory agents for the treatment of otitis media].
The objective of the present study was to estimate the therapeutic efficacy and safety of the topically applied otinum ear drops. The authors present the results of the combined treatment of acute catarrhal otitis in the children with the use of choline salicilate (otinum). The study included 50 patients randomized into two identical groups. The children of group 1 received systemic therapy supplemented by the topical application of otinum, those in group 2 were prescribed a 3% alcoholic solution of boric acid. The study has demonstrated a significantly more pronounced positive dynamics of clinical conditions in the patients of group 1 compared with those of the control group. The total duration of therapy in the first group was 37.5% shorter than in the second. The results of the study confirmed the strong anti-inflammatory and analgesic action of choline salicilate. The pain was relieved within 7 minutes on the average after the application of this agent. It is concluded that otinum can be recommended for the introduction into combined therapy of acute catarrhal otitis media as an efficacious anti-inflammatory and analgetic drug.


Retinal light-damage in albino rats: lysosomal enzymes, rhodopsin, and age.
The damaging effects incurred during a light-insult on the albino rat retina are studied separately from the resultant loss of visual cells which occurs. Groups of rats, 5-20 weeks of age, are exposed to 48 hr of 80 lux illumination, maintained in the dark for various lengths of time, and then killed. Beginning at light-off and continuing throughout 3 days of darkness, assays are made of (i) outer nuclear layer thickness; (ii) whole retina rhodopsin levels; and (iii) activities of lysosomal proteases known to be important in the degradation of retinal and ocular tissue. It is found that (a) having been given the light-insult, most of the visual cells that will die and disappear do so during the dark, postexposure period; (b) the retinas of all the animals show a limited capability for regenerating rhodopsin after the light-insult but younger animals are somewhat more successful at sustaining this than are older ones; (c) proteolytic enzyme activity is greatest in older animals which are also the ones that lose the most cells.


Gastric cancer: a primer on the epidemiology and biology of the disease and an overview of the medical management of advanced disease.
Gastric cancer is a cause of significant morbidity and cancer-related mortality worldwide. Despite recent advances in targeted therapy and understanding of the biology and development of the malignancy, progress in the treatment of gastric cancer has been limited. Most newly diagnosed patients will present with incurable disease, and have a median survival of less than 1 year. Although the disease has widespread ethnic and epidemiologic differences, medical management of gastric cancer does not distinguish among the various disease subtypes. The recent report of the ToGA phase III study has validated Her2 as a molecular target in this disease, supporting the concept that a greater understanding of the biology of gastric cancer subsets may improve treatment selection and overall outcome of individual patients. This article summarizes the epidemiology and ethnic variation of this disease to crystalize subtypes of gastric cancer in the context of current and future medical management of advanced disease.


The psychosocial profile of adolescent risk of homelessness.
To contrast the psychosocial profile of adolescents with risk factors for homelessness, identified using Chamberlain and MacKenzie's self-report scale, compared to the profiles of homeless adolescents. Multinomial logistic regression analyses were conducted contrasting profiles for (a) 137 homeless adolescents, (b) 766 secondary students reporting risk factors for homelessness, and (c) 4,844 students not reporting risks for homelessness. Fourteen percent of a representative population of at-school adolescents, from Victoria, Australia, showed elevated risk of homelessness. These adolescents showed depressive symptoms at least equivalent to homeless adolescents (RR 6.0, 95% CI: 4.9, 7.3, and RR 3.5, 95% CI: 2.1, 5.8, respectively). In multivariate analyses, homeless and at risk adolescents reported equivalent levels of family conflict, early problem behaviour and low opportunities and rewards for family involvement. Compared to adolescents not at risk, at risk adolescents were more likely to be female and to show poorer social skills/assertiveness and depressive symptoms. Compared to at risk adolescents, homeless adolescents showed additional family, school, peer and individual risks, but lower depressive symptomatology. The findings highlight the potential we have to quickly and simply detect adolescents showing significant risk of homelessness. This sizable minority of adolescents report risks often equivalent to homeless adolescents. It is hoped that stakeholders working with young people will utilise this screening potential to identify and intervene effectively with this significant subpopulation of youth, and their families, while they are still at home and school.


Predictive value of targeted proteomics for coronary plaque morphology in patients with suspected coronary artery disease.
Risk stratification is crucial to improve tailored therapy in patients with suspected coronary artery disease (CAD). This study investigated the ability of targeted proteomics to predict presence of high-risk plaque or absence of coronary atherosclerosis in patients with suspected CAD, defined by coronary computed tomography angiography (CCTA). Patients with suspected CAD (n = 203) underwent CCTA. Plasma levels of 358 proteins were used to generate machine learning models for the presence of CCTA-defined high-risk plaques or complete absence of coronary atherosclerosis. Performance was tested against a clinical model containing generally available clinical characteristics and conventional biomarkers. A total of 196 patients with analyzable protein levels (n = 332) was included for analysis. A subset of 35 proteins was identified predicting the presence of high-risk plaques. The developed machine learning model had fair diagnostic performance with an area under the curve (AUC) of 0·79 ± 0·01, outperforming prediction with generally available clinical characteristics (AUC = 0·65 ± 0·04, p < 0·05). Conversely, a different subset of 34 proteins was predictive for the absence of CAD (AUC = 0·85 ± 0·05), again outperforming prediction with generally available characteristics (AUC = 0·70 ± 0·04, p < 0·05). Using machine learning models, trained on targeted proteomics, we defined two complementary protein signatures: one for identification of patients with high-risk plaques and one for identification of patients with absence of CAD. Both biomarker subsets were superior to generally available clinical characteristics and conventional biomarkers in predicting presence of high-risk plaque or absence of coronary atherosclerosis. These promising findings warrant external validation of the value of targeted proteomics to identify cardiovascular risk in outcome studies. FUND: This study was supported by an unrestricted research grant from HeartFlow Inc. and partly supported by a European Research Area Network on Cardiovascular Diseases (ERA-CVD) grant (ERA CVD JTC2017, OPERATION). Funders had no influence on trial design, data evaluation, and interpretation.


Study of 'at risk' factors in rural under five children.
1. In the present hospital based cross sectional study, 64.97% under five children in rural area were found to be 'at risk'. 2. The most common 'at risk' factor found in these under five children was weight below 70% of the reference (39.1%) followed by acute gastroenteritis and respiratory infections (19.3%), spacing of less than 2 years (13.2%) and working mothers where the child was being looked after by a substitute (12.2%). 3. More than one 'at risk' factors were found in 43 (21.8%) children. 4. Statistically significant association was found between 'at risk' children and illiteracy and poverty.


Biopsy rate and nonalcoholic steatohepatitis (NASH) in patients with nonalcoholic fatty liver disease (NAFLD).
Background: Licensed therapies for nonalcoholic fatty liver disease (NAFLD) do not yet exist, but clinical trials are testing treatment options. Inclusion criteria often require liver biopsy showing fibrosis (F2/3) or cirrhosis (F4) and nonalcoholic steatohepatitis (NASH). However, histological criteria pose a serious obstacle for recruitment.Aims: Characterize the relevance of liver biopsies in the selection of patients with NAFLD.Methods: Patients between 2013 and 2018 with the ICD-10 code K76.0 were analyzed. Fibrosis was defined by the NASH clinical research network (CRN) fibrosis staging system, NASH by a NAFLD activity score (NAS) ≥4. Predictive factors were determined by logistic regression.Results: Liver biopsy was performed in 87/638 (13.6%) patients (49% female, age 52.5 ± 14.0, BMI 30.4 ± 5.9 kg/m2). Fibrosis stage F0/F1/F2/F3/F4 was observed in N = 7/47/7/17/9, an NAS ≥4 in N = 27. Fibrosis stage F2/F3 and F4 along with NAS ≥4 was found in 1.7% and 0.5% of cases. Liver stiffness measurement, LSM (OR 2.3 per doubling of value; CI 1.3-4.4, p = .005) and FIB-4 (OR 2.3 per doubling of value; CI 1.2-4.4, p = .012) were significant predictors for fibrosis ≥ F2. Predictive factors for NASH were not identified.Conclusion: The biopsy rate in NAFLD patients is low and fibrosis ≥ F2 along with NAS ≥4 only present in a few cases. Transient elastography and FIB-4 are useful to select patients at risk for fibrosis for liver biopsy.


A 13-question approach to resolving serological discrepancies in the transfusion medicine laboratory.
Laboratory professionals, consultants, and treating physicians may encounter discrepancies in serological testing results for numerous reasons; identifying the reason(s) for the presence of an unexpected antibody or antigen can be challenging. A question-based approach can be useful in identifying the underlying cause of the discrepancy. We describe a new approach to serological problems in a transfusion-service laboratory. The approach we outline herein is targeted towards a general transfusion medicine service, rather than a center that offers complex antibody investigations using specialized techniques. This question-based problem-solving approach considers patient factors including diagnosis, transfusion history, previous pregnancies, and medication history, along with serological test results: ABO and Rh groups, direct and indirect antiglobulin tests, reacting temperature of the antibody, effect of enzyme treatment of cells, strength of reactivity, and antibody reactivity with umbilical cord cells. We also demonstrate the usefulness of this approach through a case scenario.


ASH ISTH NHF WFH 2021 guidelines on the diagnosis of von Willebrand disease.
von Willebrand disease (VWD) is the most common inherited bleeding disorder known in humans. Accurate and timely diagnosis presents numerous challenges. These evidence-based guidelines of the American Society of Hematology (ASH), the International Society on Thrombosis and Haemostasis (ISTH), the National Hemophilia Foundation (NHF), and the World Federation of Hemophilia (WFH) are intended to support patients, clinicians, and other health care professionals in their decisions about VWD diagnosis. ASH, ISTH, NHF, and WFH established a multidisciplinary guideline panel that included 4 patient representatives and was balanced to minimize potential bias from conflicts of interest. The Outcomes and Implementation Research Unit at the University of Kansas Medical Center (KUMC) supported the guideline-development process, including performing or updating systematic evidence reviews up to 8 January 2020. The panel prioritized clinical questions and outcomes according to their importance for clinicians and patients. The panel used the Grading of Recommendations Assessment, Development and Evaluation (GRADE) approach, including GRADE Evidence-to-Decision frameworks, to assess evidence and make recommendations, which were subsequently subject to public comment. The panel agreed on 11 recommendations. Key recommendations of these guidelines include the role of bleeding-assessment tools in the assessment of patients suspected of VWD, diagnostic assays and laboratory cutoffs for type 1 and type 2 VWD, how to approach a type 1 VWD patient with normalized levels over time, and the role of genetic testing vs phenotypic assays for types 2B and 2N. Future critical research priorities are also identified.


Optimal loading of scraped HeLa cells with monoclonal antibodies to the proliferation-associated Mr 120,000 nucleolar antigen.
Our laboratory has reported (J. W. Freeman et al., Cancer Res., 48:1244-1251, 1988) a proliferation-associated Mr 120,000 nucleolar antigen (p120), that was found in human tumors but was not detectable in most normal resting tissues and in benign tumors. To study further the function and the localization of this protein, we have investigated various methods of microinjecting p120 monoclonal antibodies into cells. For comparison, we have used a monoclonal antibody to protein C23, a nucleolar protein found in high levels in most cells. To determine optimal conditions for loading of antibodies to nucleolar antigens into mechanically disrupted HeLa cells, we studied the effects of ionic strengths of loading buffers, various antibody concentrations, and optimal time for loading and antibody localization. With ascites fluids in isotonic buffer containing antibodies to nucleolar proteins p120 and C23, a maximum number of cells, 86 and 84%, respectively, were loaded following a 20-min incubation. Hypotonic buffers decreased the percentage of cells loaded (22%); hypertonic buffers reduced cell viability. The optimal concentration of purified antibody yielding a maximum number of loaded cells (81%) was 2.5 mg/ml. Higher concentrations of antibody resulted in residual cytoplasmic staining without increasing the percentage of loaded cells. With antibody concentrations less than 2.5 mg/ml, a linear decrease was noted in the percentage of cells loaded with a decrease in intensity of fluorescence. Following antibody loading, nucleolar fluorescence was observed by 12 h and the intensity increased at 24 h. Localization of the p120 antibody was followed through mitosis where it was perichromosomal and equally divided between the chromosomes at metaphase. A decrease of nucleolar immunofluorescence intensity and percentage of cells labeled were observed in successive cell generations.


A Single Cysteine Residue in the Translocation Pathway of the Mitosomal ADP/ATP Carrier from Cryptosporidium parvum Confers a Broad Nucleotide Specificity.
Cryptosporidiumparvum is a clinically important eukaryotic parasite that causes the disease cryptosporidiosis, which manifests with gastroenteritis-like symptoms. The protist has mitosomes, which are organelles of mitochondrial origin that have only been partially characterized. The genome encodes a highly reduced set of transport proteins of the SLC25 mitochondrial carrier family of unknown function. Here, we have studied the transport properties of one member of the C. parvum carrier family, demonstrating that it resembles the mitochondrial ADP/ATP carrier of eukaryotes. However, this carrier has a broader substrate specificity for nucleotides, transporting adenosine, thymidine, and uridine di- and triphosphates in contrast to its mitochondrial orthologues, which have a strict substrate specificity for ADP and ATP. Inspection of the putative translocation pathway highlights a cysteine residue, which is a serine in mitochondrial ADP/ATP carriers. When the serine residue is replaced by cysteine or larger hydrophobic residues in the yeast mitochondrial ADP/ATP carrier, the substrate specificity becomes broad, showing that this residue is important for nucleotide base selectivity in ADP/ATP carriers.


Use of NIR and pyrolysis-MBMS coupled with multivariate analysis for detecting the chemical changes associated with brown-rot biodegradation of spruce wood.
Near infrared (NIR) spectroscopy and pyrolysis-molecular beam mass spectrometry (py-MBMS) analysis can be used in conjunction with multivariate regression and principal components analysis to differentiate brown-rot-degraded wood from non-degraded spruce and to follow the temporal changes in wood undergoing brown-rot degradation. Regression of NIR test results vs. percent weight loss for Postia placenta- and Gloeophyllum trabeum-infected spruce wood blocks yielded a correlation coefficient of 0.96. Regression of MBMS test results for the same samples yielded a correlation coefficient of 0.96. Principle components analysis was used to differentiate non-infected wood and P. placenta- and G. trabeum-infected wood. These techniques may be used to detect different types of biodegradation and to develop a better understanding of the chemical changes that the wood undergoes when it is subjected to brown-rot biodegradation.


Empirical impression technique for artificial eye fitting.
Using an impression of the anophthalmic socket to facilitate the design of an artificial eye is common practice today. The Modified Impression technique was described in the American Journal of Ophthalmology, February 1969, by Lee Allen and Howard E. Webster. It is a highly successful method that involves taking an impression of the socket using an impression tray, then making a wax casting of the impression, and final modifications to the anterior aspect of the wax model. A different technique has been in use in our office and elsewhere for more than 15 years, also with a high degree of success. We've dubbed it the "Empirical/Impression" method. It involves similar steps to the Modified Impression system, but in different order: first a wax model of the anterior aspect of the eye is designed and modified; second, an impression is made of the socket, using this wax model as an impression tray. The primary advantage of this method is efficiency. It involves one less laboratory procedure, hence making a "one-day custom eye" a reality. Although this process can be used in almost any case, the Modified Impression technique may work better for certain highly irregular sockets where "reading" the fornices by empirical means may be difficult. Either method requires a highly skilled and experienced fitter to make appropriate modifications to the anterior aspect of the prosthesis.


Cochrane corner: digital contact tracing technologies in epidemics.
Contact tracing is a public health measure implemented to control the spread and break the chains of transmission of an infectious disease. It is done by identifying, assessing, and managing people who have been exposed to an infectious disease to prevent onward transmission. We summarize findings from a rapid Cochrane review that included cohort and modelling studies to assess the benefits and harms of digital solutions for identifying contacts of confirmed positive cases of an infectious disease. The review included 12 studies, which assessed digital contact tracing for the following infectious diseases: Ebola, tuberculosis, pertussis and coronavirus disease 2019 (COVID-19). This review revealed low-certainty evidence of a decrease in secondary cases of the targeted infectious disease, if digital contact tracing was used. However, it is uncertain from the currently available evidence whether digital contact tracing would produce more reliable counts of contacts and reduce the time taken to complete contact tracing. Therefore, implementation of digital contact tracing in the context of the ongoing coronavirus pandemic in African countries should be accompanied by a robust monitoring and evaluation framework. There should be an evaluation and documentation of the benefits, cost-effectiveness, acceptability, feasibility, equity impacts, and unintended consequences of the intervention.


Oestradiol implants for gender-affirming hormone therapy: an observational study of serum oestradiol levels and consumer survey.
Custom-compounded subcutaneous implants are being used widely in Australia for gender-affirming hormone therapy. However, there is no published literature regarding their use for this purpose. Electronic medical records were audited for consecutive clients who received oestradiol implants April 2019-November 2022 in gender clinics held within Hunter New England Health District in New South Wales, Australia. Serum oestradiol levels were analysed for implant doses 50-200mg, and predicted oestradiol level was modelled following 100mg implant insertion. An electronic consumer survey was sent to a convenience sample of implant recipients. A total of 38 clients received 88 implants, with 100mg oestradiol implants being the most frequently used (68%). The median interval between insertion procedures was 270 (IQR 186-399) days. The median serum oestradiol levels following implant insertion, for all implants combined, were within the target range of 250-600pmol/L at 1-, 3-, 6-, 9- and 12-month time points. Following insertion of a 100mg implant, the estimated time to reach a predicted serum oestradiol of ≤250pmol/L was 4months after an initial implant, and 13months after subsequent implants. Seventeen consumer surveys were received from 28 invitations. All respondents had previous experience of oral and/or transdermal oestradiol use. Oestradiol implants were preferred due to ease of use, perceived effectiveness, and the belief that other methods were less safe or associated with intolerance and side effects. Oestradiol implants are effective in achieving target serum oestradiol levels over a sustained period. Further research with larger cohorts could identify the optimal dosage regimen.


Stable Gene Regulatory Network Modeling From Steady-State Data.
Gene regulatory networks represent an abstract mapping of gene regulations in living cells. They aim to capture dependencies among molecular entities such as transcription factors, proteins and metabolites. In most applications, the regulatory network structure is unknown, and has to be reverse engineered from experimental data consisting of expression levels of the genes usually measured as messenger RNA concentrations in microarray experiments. Steady-state gene expression data are obtained from measurements of the variations in expression activity following the application of small perturbations to equilibrium states in genetic perturbation experiments. In this paper, the least absolute shrinkage and selection operator-vector autoregressive (LASSO-VAR) originally proposed for the analysis of economic time series data is adapted to include a stability constraint for the recovery of a sparse and stable regulatory network that describes data obtained from noisy perturbation experiments. The approach is applied to real experimental data obtained for the SOS pathway in Escherichia coli and the cell cycle pathway for yeast Saccharomyces cerevisiae. Significant features of this method are the ability to recover networks without inputting prior knowledge of the network topology, and the ability to be efficiently applied to large scale networks due to the convex nature of the method.


Presentation and management of arthrofibrosis of the knee: A case report.
Case report. Arthrofibrosis is a debilitating condition that results in pain, decreased range of motion, and decreased function. Although surgical management of arthrofibrosis has been well described in the literature, rehabilitation of the arthrofibrotic knee is less well described. A 28-year-old female presented with swelling, pain, and decreased strength, range of motion, patellar mobility, and function following an exploratory arthroscopy of her left knee. After failed conservative management, the patient underwent two additional surgeries to remove scar tissue. Following each surgery, the emphasis was on decreasing inflammation and maintaining patellar mobility while increasing joint range of motion and strength. Therapy progression was determined by the presence or absence of inflammatory signs. The second scar tissue removal surgery resulted in a femoral neuropathy that further complicated the rehabilitation process. At 3-year follow-up, the patient continued to present with decreased range of motion and strength compared to the uninvolved limb, but had returned to a modified running program and reported pain no longer limited her ability to participate in activities of daily living. This case report highlights the importance of recognizing that arthrofibrosis may result following a minor knee surgery and with minimal range of motion loss. Additional complications also may result during arthrofibrosis treatment. Progressing rehabilitation based on the inflammatory response may decrease the likelihood of additional scar tissue formation and potentially improve the outcome for the patient.


Control of her1 expression during zebrafish somitogenesis by a delta-dependent oscillator and an independent wave-front activity.
Somitogenesis has been linked both to a molecular clock that controls the oscillation of gene expression in the presomitic mesoderm (PSM) and to Notch pathway signaling. The oscillator, or clock, is thought to create a prepattern of stripes of gene expression that regulates the activity of the Notch pathway that subsequently directs somite border formation. Here, we report that the zebrafish gene after eight (aei) that is required for both somitogenesis and neurogenesis encodes the Notch ligand DeltaD. Additional analysis revealed that stripes of her1 expression oscillate within the PSM and that aei/DeltaD signaling is required for this oscillation. aei/DeltaD expression does not oscillate, indicating that the activity of the Notch pathway upstream of her1 may function within the oscillator itself. Moreover, we found that her1 stripes are expressed in the anlage of consecutive somites, indicating that its expression pattern is not pair-rule. Analysis of her1 expression in aei/DeltaD, fused somites (fss), and aei;fss embryos uncovered a wave-front activity that is capable of continually inducing her1 expression de novo in the anterior PSM in the absence of the oscillation of her1. The wave-front activity, in reference to the clock and wave-front model, is defined as such because it interacts with the oscillator-derived pattern in the anterior PSM and is required for somite morphogenesis. This wave-front activity is blocked in embryos mutant for fss but not aei/DeltaD. Thus, our analysis indicates that the smooth sequence of formation, refinement, and fading of her1 stripes in the PSM is governed by two separate activities.


Does diabetes protect the femoral artery against atherosclerosis?
To compare the extent of angiographically detected stenoses in different segments of the lower leg arteries in diabetics and nondiabetics. The study group consisted of 60 nondiabetic and 38 diabetic patients with lower extremity disease and no previous vascular interventions. Arterial lesions were analysed by digital subtraction angiography (DSA). The patients were evaluated by walking distance and ankle-brachial index (ABI) and the reason for hospitalization was recorded. Statistically significantly fewer stenoses were found in the superficial femoral artery in diabetics than in nondiabetics. Although a tendency to more diffuse changes in crural arteries was found, no statistically significant difference were found in other segments of the lower leg arteries apart from the femoral arteries. Judging from the present results there are fewer angiographically detectable stenoses in the superficial arteries in diabetics than nondiabetics.


Bioavailability of transdermal methimazole in a pluronic lecithin organogel (PLO) in healthy cats.
The antithyroid drug methimazole is widely used for the medical management of feline hyperthyroidism. Recently, custom veterinary pharmacies have offered methimazole in a transdermal gel containing pluronic and lecithin (PLO), with anecdotal evidence of efficacy. The purpose of this study was to determine the bioavailability, relative to i.v. and oral routes of administration, of transdermal methimazole in a PLO gel in cats. Six healthy adult cats were assigned to receive 5 mg of methimazole by the i.v., oral, or transdermal routes, in a randomized triple crossover protocol with 1 week washout between doses. Blood samples were taken for high performance liquid chromatography (HPLC) determination of serum methimazole, at 0, 5, 15, 30, 60 min, and 2, 4, 6, 12 and 24 h after dosing. Methimazole absorption following transdermal administration was poor and variable, with only two of six cats achieving detectable serum methimazole concentrations at any time point following transdermal administration. Area under the concentration-time curve (AUC), maximum concentration (Cmax), and absolute bioavailability were all significantly lower for the transdermal route (0.39 +/- 0.63 microg h/mL, 0.05 +/- 0.09 microg/mL, and 11.4 +/- 18.7%, respectively) than for either i.v. (7.96 +/- 4.38 microg h/mL, 3.34 +/- 2.00 microg/mL, 100%) or oral routes (2.94 +/- 1.24 microg h/mL, 0.51 +/- 0.15 microg/mL, 40.4 +/- 8.1%). The results of this study indicate generally low to undetectable bioavailability of methimazole in a lecithin/pluronic gel given as a single transdermal dose to healthy cats, although one individual cat did achieve nearly 100% transdermal bioavailability relative to the oral route.


Development and validation of measure of household food insecurity in urban Costa Rica confirms proposed generic questionnaire.
Interest in household food insecurity (FI) within scientific and policy groups has motivated efforts to develop methods for measuring it. Questionnaires asking about FI experiences have been shown to be valid in the contexts in which they were created. The issue has arisen as to whether such questionnaires need be developed from the ground up or if a generic questionnaire can be adapted to a particular context. This study aimed to gain an in-depth understanding of household FI in urban Costa Rica, develop and validate a questionnaire for its measurement, and inform the choice between the 2 methods of development. The study was conducting using qualitative and quantitative methods provided in the Food and Nutrition Technical Assistance (FANTA) guidelines. In-depth interviews were conducted with 49 low-middle-income urban women using a semistructured interview guide. A 14-item FI questionnaire was developed based on results from these interviews. A field study was conducted in 213 households. The results show that the developed questionnaire provides valid measurement of household FI in urban Costa Rica and is simple and quick to apply in the household setting. FANTA developed a guide during the period that this research was completed that provides a generic questionnaire that can be adapted for use in various countries, rather than building the questionnaire from the ground up. This study provides evidence that careful attention to the procedures in this guide will likely yield a questionnaire suitable for assessing household FI in middle-income countries.


Evaluation of 99mTc labelled poly lactic acid microspheres for diagnostic radioembolization.
Radioembolization is used in diagnostic imaging of the lungs and for radioembolization therapy of hepatic tumours. Presently, 99mTc labelled macroaggregates or microspheres of human serum albumin (HAM) are used for this purpose. Poly lactic acid (PLA) is biodegradable, like HAM, and, unlike HAM, is not a blood product. The aim of the present study was to evaluate the uptake and biodegradation of PLA microspheres in lungs. PLA (MW = 48720 Da) microspheres of 1.0-100 microm (mean = 39.5 microm) in diameter were prepared by solvent evaporation from methylene chloride. They were labelled with 99mTc by stannous chloride reduction at pH 3, with an efficiency of 98% and a stability of 96% at 24 h. For biodistribution studies, 15 mice were i.v. injected with 20 microCi 99mTc-PLA microspheres in 0.1 ml and sacrificed at 15 min, 1, 3, 6 and 24 h (three at each time point). All the organs were removed, weighed and counted against a standard prepared from 1/100 dilution of the injected radioactivity. Some mice were similarly injected and sacrificed at 30 min, 15 and 30 days. The lungs were removed and frozen, and 10 microm sections were obtained, stained with haemotoxylin and eosin and examined under a light microscope. Five rabbits were i.v. injected with 1 mCi of 99mTc-PLA microspheres. Scintigrams were obtained at various intervals up to 24 h. In mice, the lung uptake was significant at 30 min-1h post-injection. In rabbits, the lungs were the only organs visualized up to 24 h. Microscopic examination of tissue sections demonstrated slow biodegradation of PLA particles. In conclusion; (1) The high lung uptake obtained in mice and rabbits indicates the suitability of PLA microspheres for lung imaging, and (2) although the slow biodegradation rate might be a disadvantage in patients with lung disorders in diagnostic studies, it may be an advantage in therapeutic applications with radionuclides which have long physical half lives.


Increased hepatic expression is a major determinant of serum alanine aminotransferase elevation in mice with nonalcoholic steatohepatitis.
Serum alanine aminotransferase (ALT) is a biomarker for hepatitis of various aetiologies including fatty liver disease. Increased serum ALT is thought to be related to its increased release from dying hepatocytes. We sought to understand the mechanisms by which serum ALT is elevated in a mouse model of experimental fatty liver disease where hepatocyte death is minimal. To induce fatty liver disease, female A/J mice were fed a methionine-choline deficient (MCD) diet for up to 12 weeks. Serum and liver ALT expression and hepatic inflammation, necrosis and apoptosis were assessed and expressed relative to their expressions in control-diet-fed mice. Feeding mice the MCD diet produced hepatic steatosis with minimal hepatic inflammation or necrosis. Liver cell apoptosis was not significantly increased by MCD diet treatment. Conversely, serum ALT activity was approximately four-fold increased at 12 weeks of diet treatment, and ALT protein expressions in serum were correspondingly increased: ALT1 1.7-fold and ALT2 1.9-fold at 12 weeks. The expressions of ALT1 and ALT2 protein in liver increased over 2-12 weeks of MCD treatment. At 12 weeks, liver ALT1 protein was 2.27+/-0.31-fold increased and ALT2 protein 4.72+/-0.48-fold increased relative to their expressions in the mice fed a diet replete with methionine and choline. Liver ALT mRNA expressions were correspondingly increased: ALT1 mRNA 2.58-fold and ALT2 mRNA 4.97-fold at 12 weeks. Linear regression analysis showed a strong correlation between serum and liver tissue expressions for both ALT1 and ALT2. These findings suggest that induction of hepatic expression significantly contributes to increased serum ALT in this model of experimental fatty liver disease, whereas cell death appears not to.


Interleukin-1beta expression and phospholipase A(2) activation after intestinal ischemia/reperfusion injury.
The experiments were carried out to explore the interactions between IL-1 beta gene expression, protein level and phospholipase A(2) PLA(2) inhibition after intestinal ischemia/reperfusion injury. Using a rat intestinal ischemia/reperfusion injury model, after collecting the serum, lung lavage, abdomen cavity lavage and important organ tissue samples from control, injury and PLA(2) inhibitor treated groups, IL-1 beta level was measured by radioimmunoassay, and the mRNA expression of IL-1 beta and type II PLA (2)was determined by RT-PCR. After 6 h of injury, the IL-1 beta level in serum was significantly higher than that in the control group; an increase in IL-1 beta was also observed in abdomen cavity lavage 1 or 3 h after injury. IL-1 beta was significantly increased in liver tissue after injury, but was not changed obviously in the lung, kidney and intestinal tissues. IL-1 beta in the lung lavage was significantly higher than that of control group. The mRNA expression of IL-1 beta in lung tissue was increased after injury, but type II PLA(2) mRNA expression was decreased. There were different changes in IL-1 beta level and gene expression after treatment with PLA(2) inhibitor chloroquine, cyclo-oxidase inhibitor indomethacin, or PAF receptor antagonist SR27417 respectively after injury. All these results indicate that after intestinal ischemia/reperfusion injury, the IL-1 beta level and mRNA gene expression are significantly increased, however, the relationship among IL-1 beta, PLA(2) activation and its metabolite release remains to be further elucidated.


Carnitine acetyltransferase (Crat) in hunger-sensing AgRP neurons permits adaptation to calorie restriction.
Hunger-sensing agouti-related peptide (AgRP) neurons ensure survival by adapting metabolism and behavior to low caloric environments. This adaption is accomplished by consolidating food intake, suppressing energy expenditure, and maximizing fat storage (nutrient partitioning) for energy preservation. The intracellular mechanisms responsible are unknown. Here we report that AgRP carnitine acetyltransferase (Crat) knockout (KO) mice exhibited increased fatty acid utilization and greater fat loss after 9 d of calorie restriction (CR). No differences were seen in mice with ad libitum food intake. Eleven days ad libitum feeding after CR resulted in greater food intake, rebound weight gain, and adiposity in AgRP Crat KO mice compared with wild-type controls, as KO mice act to restore pre-CR fat mass. Collectively, this study highlights the importance of Crat in AgRP neurons to regulate nutrient partitioning and fat mass during chronically reduced caloric intake. The increased food intake, body weight gain, and adiposity in KO mice after CR also highlights the detrimental and persistent metabolic consequence of impaired substrate utilization associated with CR. This finding may have significant implications for postdieting weight management in patients with metabolic diseases.-Reichenbach, A., Stark, R., Mequinion, M., Lockie, S. H., Lemus, M. B., Mynatt, R. L., Luquet, S., Andrews, Z. B. Carnitine acetyltransferase (Crat) in hunger-sensing AgRP neurons permits adaptation to calorie restriction.


The Interactive Management of the SARS-CoV-2 Virus: The Social Cohesion Index, a Methodological-Operational Proposal.
This contribution places itself within the emergency context of the COVID-19 spread. Until medical research identifies a cure acting at an organic level, it is necessary to manage what the emergency generates among the members of the Community in interactive terms in a scientific and methodologically well-founded way. This is in order to promote, among the members of the Community, the pursuit of the common aim of reducing the spread of infection, with a view to community health as a whole. In addition, being at the level of interactions enables us to move towards a change of these interactions in response to the COVID-19 emergency, in order to manage what will happen in the future, in terms of changes in the interactive arrangements after the emergency itself. This becomes possible by shifting away from the use of deterministic-causal references to the use of the uncertainty of interaction as an epistemological foundation principle. Managing the interactive (and non-organic) fallout of the emergency in the Community is made possible by the formalisation of the interactive modalities (the Discursive Repertories) offered by Dialogical Science. To place oneself within this scientific panorama enables interaction measurements: so, the interaction measurement indexes offers a range of generative possibilities of realities built by the speeches of the Community members. Moreover, the Social Cohesion measurement index, in the area of Dialogical Science, makes available to public policies the shared measure of how and by how much the Community is moving towards the common purpose of reducing the contagion spread, rather than moving towards other personal and not shared goals (for instance, having a walk in spite of the lockdown). In this index, the interaction between the Discursive Repertories and the "cohesion weight" associated with them offers a Cohesion output: the data allow to manage operationally what happens in the Community in a shared way and in anticipation, without leaving the interactions between its members to chance. In this way, they can be directed towards the common purpose through appropriate interventions relevant to the interactive set-up described in the data. The Cohesion measure makes it possible to operate effectively and efficiently, thanks to the possibility of monitoring the progress of the interventions implemented and evaluating their effectiveness. In addition, the use of predictive Machine Learning models, applied to interactive cohesion data, allows for immediate and efficient availability of the measure itself, optimising time and resources.


Moderate to severe psoriasis: a single-center analysis of gender prevalence.
Psoriasis is a chronic, relapsing disease and most epidemiological studies include selected patients undergoing systemic therapies only. Epidemiological data suggest that psoriasis affects 2-3% of the general population, and that men and women are equally affected. The objective was to identify differences in gender for disease severity, patient characteristics and comorbidities in patients with moderate to severe psoriasis, independent of therapy. A retrospective medical chart review of consecutive patients diagnosed with moderate-severe psoriasis at a single center between 2004 and 2017, with a complete set of medical records, was undertaken. Both univariate and multivariate regression analyses were performed. Statistical significance was defined as P<0.05. The male-to-female ratio revealed a higher prevalence for male gender (2:1, P<0.05). Whilst no significant differences were found for most factors according to gender, age at first evaluation was significantly higher for women. Logistic regression analysis indicated that autoimmune/autoinflammatory diseases were more frequently observed in women, as well as phenotypes other than plaque psoriasis and hypertension. Inversely, dyslipidemia was more frequently associated with male gender. Our results show that moderate-severe psoriasis is more common in men and suggests a differential gender distribution of some specific comorbidities in the setting of this disease.


Cognitive outcome following staged bilateral pallidal stimulation for the treatment of Parkinson's disease.
As neurosurgical treatment of parkinsonian symptoms has become increasingly popular, concern about the cognitive morbidity which may result from such interventions has risen proportionately. Previous reports of cognitive difficulties associated with pallidotomy and thalamotomy, especially in bilateral cases, have provided the impetus for research into chronic electrical deep brain stimulation procedures which are believed to be safer than ablation. Given the lack of neurobehavioral research following bilateral deep brain stimulation procedures, this preliminary study of six Parkinson's disease patients undergoing staged bilateral pallidal stimulation was undertaken. A battery of tests assessing attention, executive function, visuomotor coordination, language, visuoperceptual function, learning memory and mood revealed no significant change in overall level of cognitive functioning after either unilateral or bilateral pallidal deep brain stimulation. No significant declines were observed about three months following bilateral stimulation, and in fact, significant gains in delayed recall and relief of anxiety symptoms were noted. It was concluded from this preliminary data that bilateral pallidal stimulation for the treatment of Parkinson's disease, at least in the absence of operative complications, offers a cognitively safe alternative to ablation.


You Cannot Step Into the Same River Twice: When Power Analyses Are Optimistic.
Statistical power depends on the size of the effect of interest. However, effect sizes are rarely fixed in psychological research: Study design choices, such as the operationalization of the dependent variable or the treatment manipulation, the social context, the subject pool, or the time of day, typically cause systematic variation in the effect size. Ignoring this between-study variation, as standard power formulae do, results in assessments of power that are too optimistic. Consequently, when researchers attempting replication set sample sizes using these formulae, their studies will be underpowered and will thus fail at a greater than expected rate. We illustrate this with both hypothetical examples and data on several well-studied phenomena in psychology. We provide formulae that account for between-study variation and suggest that researchers set sample sizes with respect to our generally more conservative formulae. Our formulae generalize to settings in which there are multiple effects of interest. We also introduce an easy-to-use website that implements our approach to setting sample sizes. Finally, we conclude with recommendations for quantifying between-study variation.


Divergence of premating behaviors in the closely related species Drosophila subquinaria and D. recens.
Most animal species use distinctive courship patterns to choose among potential mates. Over time, the sensory signaling and preferences used during courtship can diverge among groups that are reproductively isolated. This divergence of signal traits and preferences is thought to be an important cause of behavioral isolation during the speciation process. Here, we examine the sensory modalities used in courtship by two closely related species, Drosophila subquinaria and Drosophila recens, which overlap in geographic range and are incompletely reproductively isolated. We use observational studies of courtship patterns and manipulation of male and female sensory modalities to determine the relative roles of visual, olfactory, gustatory, and auditory signals during conspecific mate choice. We find that sex-specific, species-specific, and population-specific cues are used during mate acquisition within populations of D. subquinaria and D. recens. We identify shifts in both male and female sensory modalities between species, and also between populations of D. subquinaria. Our results indicate that divergence in mating signals and preferences have occurred on a relatively short timescale within and between these species. Finally, we suggest that because olfactory cues are essential for D. subquinaria females to mate within species, they may also underlie variation in behavioral discrimination across populations and species.


Mutations of ovine and bovine placental lactogens change, in different ways, the biological activity mediated through homologous and heterologous lactogenic receptors.
The biological activities of ovine (o) and bovine (b) placental lactogens (PLs) and their mutated analogues were compared using several binding and in vitro bioassays. In almost all cases, the biological activities of these analogues mediated through rat (r) prolactin receptor (PRLR) showed little or no change, despite a remarkable decrease in their capacity to bind to the extracellular domain of rPRLR and despite compromised stability of the 2:1 complexes. These results indicate that mutations impairing the ability of oPL or bPL to form stable complexes with lactogenic receptors do not necessarily lead to a decrease in the biological activity, because the transient existence of the homodimeric complex is still sufficient to initiate the signal transduction. In contrast, oPL and bPL analogues completely, or almost completely, lost their ability to activate homologous PRLRs, and some of them even acted as site-2 antagonists. To explain the difference between the activity transduced through homologous and that transduced through heterologous PRLRs, we propose the novel term 'minimal time of homodimer persistence'. This concept assumes that in order to initiate the signal transduction, the associated kinase JAK2 has to be transphosphorylated and this requires a 'minimal time' of homodimer existence. In the case of homologous interaction between ruminant PLs and homologous PRLRs, this 'minimal time' is met, though the interaction with homologous PRLRs has a shorter half-life than that with heterologous PRLRs. Therefore oPL or bPL are active in cells possessing both homologous and heterologous PRLRs. Mutations of oPL or bPL lead to reduced affinity and, consequently, the 'time of homodimer persistence' is shortened. Although in the case of heterologous interaction the 'minimal time' is still sufficient to initiate the biological activity, in homologous interactions, which are already weaker than heterologous interactions, further destabilization of the complex shortens its persistence to below the 'minimal time', leading to full or partial loss of biological activity.


An unusual spliced variant of DELLA protein, a negative regulator of gibberellin signaling, in lettuce.
DELLA proteins are negative regulators of the signaling of gibberellin (GA), a phytohormone regulating plant growth. DELLA degradation is triggered by its interaction with GID1, a soluble GA receptor, in the presence of bioactive GA. We isolated cDNA from a spliced variant of LsDELLA1 mRNA in lettuce, and named it LsDELLA1sv. It was deduced that LsDELLA1sv encodes truncated LsDELLA1, which has DELLA and VHYNP motifs at the N terminus but lacks part of the C-terminal GRAS domain. The recombinant LsDELLA1sv protein interacted with both Arabidopsis GID1 and lettuce GID1s in the presence of GA. A yeast two-hybrid assay suggested that LsDELLA1sv interacted with LsDELLA1. The ratio of LsDELLA1sv to LsDELLA1 transcripts was higher in flower samples at the late reproductive stage and seed samples (dry seeds and imbibed seeds) than in the other organ samples examined. This study suggests that LsDELLA1sv is a possible modulator of GA signaling in lettuce.


Expansion of an FMR1 grey-zone allele to a full mutation in two generations.
Fragile X Syndrome is caused by the expansion of an unstable CGG-repeat tract in the 5'-UTR of the FMR1 gene, which generally results in transcriptional silencing and consequent absence of the FMR1 protein. To date, the smallest premutation allele reported to expand to a full mutation allele in a single generation is 59 CGG repeats. Here, we report a single-generation expansion to a full mutation allele (male with approximately 538 CCG repeats) from a mother who is a carrier of a premutation allele of 56 CGG repeats. Furthermore, the maternal grandfather was a carrier of a gray (or intermediate)-zone allele (45 to 54 repeats) of 52 CGG repeats. Thus, in this family, a gray-zone allele expanded to the full mutation range in two generations. Interestingly, the two AGG interruptions present in the grandfather's allele were absent in the mother's premutation allele. These observations underscore the need to consider carriers of alleles of greater than 55 CGG repeats as being at risk for transmission of a full mutation allele in a single generation, and those with even smaller alleles in the gray zone as being at risk of having grandchildren with full mutation alleles.


Cerebral embolism from left atrial myxoma leading to cerebral and retinal aneurysms: a case report.
Left atrial myxoma commonly leads to cerebral embolic ischemic stroke. The subsequent formation of cerebral aneurysms due to myxomatous emboli is a phenomenon that may lead to severe neurologic complications such as intracerebral hemorrhage. In addition to the formation of aneurysms in cerebral arteries, we report here the unique picture of a retinal involvement consisting in microaneurysm formation associated with myxomatous embolism.


COVID-19 Lessons Learned: Response to the Anticipated Ventilator Shortage.
Early in the COVID-19 pandemic predictions of a worldwide ventilator shortage prompted a worldwide search for solutions. The impetus for the scramble for ventilators was spurred on by inaccurate and often unrealistic predictions of ventilator requirements. Initial efforts looked simply at acquiring as many ventilators as possible from national and international sources. Ventilators from the Strategic National Stockpile were distributed to early hotspots in the Northeast and Northwest United States. In a triumph of emotion over logic, well-intended experts from other industries turned their time, talent, and treasure toward making a ventilator for the first time. Interest in shared ventilation (more than one patient per ventilator) was ignited by an ill-advised video on social media that ignored the principles of gas delivery in deference to social media notoriety. With shared ventilation, a number of groups mistook a physiologic problem for a plumbing problem. The United States government invoked the Defense Production Act to push automotive manufacturers to partner with existing ventilator manufacturers to speed production. The FDA granted emergency use authorization for "splitters" to allow shared ventilation as well as for ventilators and ancillary equipment. Rationing of ventilators was discussed in the lay press and medical literature but was never necessary in the US. Finally, planners realized that staff with expertise in providing mechanical ventilation were the most important shortage. Over 200,000 ventilators were purchased by the United States government, states, cities, health systems, and individuals. Most had little value in caring for patients with COVID-19 ARDS. This paper attempts to look at where miscalculations were made, with an eye toward what we can do better in the future.


Detection of prostate cancer and predicting progression: current and future diagnostic markers.
Carcinoma of the prostate is the second leading cause of male cancer-related death in the United States. Better indicators of prostate cancer presence and progression are needed to avoid unnecessary treatment, predict disease course, and develop more effective therapy. Numerous molecular markers have been described in human serum, urine, seminal fluid, and histological specimens that exhibit varying capacities to detect prostate cancer and predict disease course. However, to date, few of these markers have been adequately validated for clinical use. The purpose of this review is to examine the current status of these markers in prostate cancer and to assess the diagnostic potential for future markers from identified genes and molecules that display loss, mutation, or alteration in expression between tumor and normal prostate tissues. In this review we cite 91 molecular markers that display some level of correlation with prostate cancer presence, disease progression, cancer recurrence, prediction of response to therapy, and/or disease-free survival. We suggest criteria to consider when selecting a marker for further development as a clinical tool and discuss five examples of markers (chromogranin A, glutathione S-transferase pi 1, prostate stem cell antigen, prostate-specific membrane antigen, and telomerase reverse transcriptase) that fulfill some of these criteria. Finally, we discuss how to conduct evaluations of candidate prostate cancer markers and some of the issues involved in the validation process.


Geographical patterns and predictors of malaria risk in Zambia: Bayesian geostatistical modelling of the 2006 Zambia national malaria indicator survey (ZMIS).
The Zambia Malaria Indicator Survey (ZMIS) of 2006 was the first nation-wide malaria survey, which combined parasitological data with other malaria indicators such as net use, indoor residual spraying and household related aspects. The survey was carried out by the Zambian Ministry of Health and partners with the objective of estimating the coverage of interventions and malaria related burden in children less than five years. In this study, the ZMIS data were analysed in order (i) to estimate an empirical high-resolution parasitological risk map in the country and (ii) to assess the relation between malaria interventions and parasitaemia risk after adjusting for environmental and socio-economic confounders. The parasitological risk was predicted from Bayesian geostatistical and spatially independent models relating parasitaemia risk and environmental/climatic predictors of malaria. A number of models were fitted to capture the (potential) non-linearity in the malaria-environment relation and to identify the elapsing time between environmental effects and parasitaemia risk. These models included covariates (a) in categorical scales and (b) in penalized and basis splines terms. Different model validation methods were used to identify the best fitting model. Model-based risk predictions at unobserved locations were obtained via Bayesian predictive distributions for the best fitting model. Model validation indicated that linear environmental predictors were able to fit the data as well as or even better than more complex non-linear terms and that the data do not support spatial dependence. Overall the averaged population-adjusted parasitaemia risk was 20.0% in children less than five years with the highest risk predicted in the northern (38.3%) province. The odds of parasitaemia in children living in a household with at least one bed net decreases by 40% (CI: 12%, 61%) compared to those without bed nets. The map of parasitaemia risk together with the prediction error and the population at risk give an important overview of the malaria situation in Zambia. These maps can assist to achieve better resource allocation, health management and to target additional interventions to reduce the burden of malaria in Zambia significantly. Repeated surveys will enable the evaluation of the effectiveness of on-going interventions.


[Family formation by schizophrenic and manic-depressive patients according to an epidemiologic study (birth rate)].
The birth rate among patients with endogenous disorders is considered as an index of the degree of their social adaptation. It was established that in all forms of such disorders the level of birth rates and the rate of family formation is lower than that in the general population. However, in all endogenous disorders (except the malignant schizophrenia) there are some patients the adaptation of whom is not disturbed.


Cyclic di-GMP stimulates protective innate immunity in bacterial pneumonia.
Innate immunity is the primary mechanism by which extracellular bacterial pathogens are effectively cleared from the lung. We have previously shown that cyclic di-GMP (c-di-GMP [c-diguanylate]) is a novel small molecule immunomodulator and immunostimulatory agent that triggers protective host innate immune responses. Using a murine model of bacterial pneumonia, we show that local intranasal (i.n.) or systemic subcutaneous (s.c.) administration of c-di-GMP prior to intratracheal (i.t.) challenge with Klebsiella pneumoniae stimulates protective immunity against infection. Specifically, i.n. or s.c. administration of c-di-GMP 48 and 24 h prior to i.t. K. pneumoniae challenge resulted in significantly increased survival. Pretreatment with c-di-GMP resulted in a 5-fold reduction in bacterial CFU in the lung (P < 0.05) and an impressive >1,000-fold decrease in CFU in the blood (P < 0.01). c-di-GMP administration stimulated a robust innate response to bacterial challenge, characterized by enhanced accumulation of neutrophils and alphabeta T cells, as well as activated NK and alphabeta T lymphocytes, which was associated with earlier and more vigorous expression of chemokines and type I cytokines. Moreover, lung macrophages recovered from Klebsiella-infected mice pretreated with c-di-GMP expressed greater quantities of inducible nitric oxide synthase and nitric oxide ex vivo than did macrophages isolated from infected mice pretreated with the control, c-GMP. These findings demonstrate that c-di-GMP delivered in either a compartmentalized or systemic fashion stimulates protective innate immunity in the lung and protects mice against bacterial invasion. We propose that the cyclic dinucleotide c-di-GMP may be used clinically as an effective immunomodulator, immune enhancer, and vaccine adjuvant to protect against respiratory infection and pneumonia in humans and animals.


Synthesis of new (-)-bestatin-based inhibitor libraries reveals a novel binding mode in the S1 pocket of the essential malaria M1 metalloaminopeptidase.
The malarial PfA-M1 metallo-aminopeptidase is considered a putative drug target. The natural product dipeptide mimetic, bestatin, is a potent inhibitor of PfA-M1. Herein we present a new, efficient, and high-yielding protocol for the synthesis of bestatin derivatives from natural and unnatural N-Boc-d-amino acids. A diverse library of bestatin derivatives was synthesized with variants at the side chain of either the α-hydroxy-β-amino acid (P1) or the adjacent natural α-amino acid (P1'). Surprisingly, we found that extended aromatic side chains at the P1 position resulted in potent inhibition against PfA-M1. To understand these data, we determined the X-ray cocrystal structures of PfA-M1 with two derivatives having either a Tyr(OMe) 15 or Tyr(OBzl) 16 at the P1 position and observed substantial inhibitor-induced rearrangement of the primary loop within the PfA-M1 pocket that interacts with the P1 side chain. Our data provide important insights for the rational design of more potent and selective inhibitors of this enzyme that may eventually lead to new therapies for malaria.


Deletion of the CYP2D6 gene as a likely explanation for the serious side effects of the antipsychotic drug pimozide: a case report.
CYP2D6 analysis prior to the prescription of pimozide is required above a certain dose by the Food and Drug Administration in order to detect individuals with the poor metabolizer status. This precautionary measure aims to prevent the occurrence of serious adverse drug reactions. This study presents a case of a patient diagnosed with schizophrenia spectrum disorder. The patient suffered re-admission in the psychiatry ward because of severe secondary symptoms due to the antipsychotic drug pimozide, previously prescribed on a first admission. In order to assess the patient's medication profile, real-time PCR was performed to analyze the main genes responsible for its metabolization, namely, CYP2D6 and CYP3A4. The pharmacogenetic study revealed that the patient is a poor metabolizer for CYP2D6, presenting deletion of both copies of the gene (diplotype *5/*5). Fortunately, the symptomatology disappeared after the withdrawal of the responsible drug. In conclusion, abiding by the pharmacogenetic clinical practice guidelines and the pharmacogenetic analysis of CYP2D6 when prescribing pimozide would have probably saved the patient from the consequences of severe side effects and the health system expenditure. There is an important need for more training in the pharmacogenetic field for specialists in psychiatry.


[A comparison between antigliadin antibodies and blood xylose in celiac disease].
We have compared serum antigliadin antibodies (AGA) with xylose absorption test in diagnosis and follow-up of pediatric celiac disease. Three groups of children were investigated: celiacs, affected by other gastrointestinal disease, healthy controls. On gluten diet AGA IgA, IgG and xylose test were abnormal in all celiac children. After only three months of gluten-free diet, abnormal AGA IgA values were found in 3%, AGA IgG in 63%, xylose test in 28% of children. Normal values for AGA IgA and IgG and for xylose test were found between 7 and 20 months. On challenge, after 1-4 months of gluten diet, abnormal AGA IgA and IgG values were found in 90% of cases, xylose test only in 27%. As far as the children with other gastrointestinal disease are concerned, 2% had abnormal values for AGA IgA, 22% for AGA IgG and 42% for xylose test. All healthy children had normal AGA IgA, IgG values and xylose test. Our date show AGA IgA the most specific laboratory test, among these investigated, for diagnosis and follow-up of celiac disease.


Nursing care of patients in intermaxillary fixation.
Often patients in IMF are healthier, tend to be more independent, and have shorter hospital stays than many patients hospitalized for other reasons. This is by no means any indication that they do not require close assessment and skilled nursing care. While recovering from anesthesia, airway obstruction and vomiting are real dangers. Although wire cutters are a necessary safety precaution and a part of the legal standard of practice, nurses and patients should not rely solely on them in the incidence of shortness of breath or vomiting. Nurses and patients should know the proper technique for clearing the mouth and airway of a patient in IMF. Hemorrhage evaluation and secretion control also require skilled observation and assistance by the nurse. Patients in IMF benefit from a decrease in anxiety levels when they are educated in airway clearance, secretion control, comfort measures, and diet. The nursing care of patients in IMF is rewarding and challenging for many areas of nursing.


Exposure to perfluorooctanoic acid leads to promotion of pancreatic cancer.
Pancreatic cancer is the fourth leading cause of cancer deaths in the United States. Perfluorooctanoic acid (PFOA), a persistent environmental pollutant, has been shown to induce pancreatic acinar cell tumors in rats. Human epidemiologic studies have linked PFOA exposure to adverse chronic health effects including several types of cancer. Previously, we demonstrated that PFOA induces oxidative stress and focal ductal hyperplasia in the mouse pancreas. Here, we evaluated whether PFOA promotes pancreatic cancer using the LSL-KRasG12D;Pdx-1 Cre (KC) mouse model of pancreatic cancer. KC mice were exposed to 5 ppm PFOA in drinking water starting at 8 weeks of age and analyzed at 6 and 9 months of age. At the 6-month time point, PFOA exposure increased pancreatic intraepithelial neoplasia (PanIN) area by 58%, accompanied by a 2-fold increase in lesion number. Although PanIN area increased at 9 months, relative to 6 months, no treatment effect was observed. Collagen deposition was enhanced by PFOA at both the 6- and 9-month time points. PFOA also induced oxidative stress in the pancreas evidenced by elevated antioxidant activity of superoxide dismutase (Sod), catalase and thioredoxin reductase, and a ~3-fold increase in Sod1 mRNA and protein levels at 6 months. Although antioxidant activity was not enhanced by PFOA exposure at the 9-month time point, increased pancreatic oxidative damage was observed. Collectively, these results show that PFOA elicited temporal increases in PanIN lesion area and desmoplasia concomitant with the induction of oxidative stress, demonstrating that it functions to promote pancreatic cancer progression.


Perceived Stigma as a Contextual Barrier to Early Uptake of HIV Testing, Treatment Initiation, and Disclosure; the Case of Patients Admitted with AIDS-Related Illness in a Rural Hospital in South Africa.
We explored the extent to which perceived HIV-related stigma influences the disclosure and concealment of HIV status to family among adult patients hospitalised for AIDS-related illness, and described reports of negative responses and enacted stigma following disclosure. We conducted interviews with a purposeful sample of 28 adult patients in a rural South African hospital. Data analysis was deductive and inductive and followed the thematic approach. We found evidence of delayed HIV diagnosis and initiation of treatment. There was delayed and selective disclosure as well as concealment of the HIV-positive status. The disclosure was delayed for months or even years. During that time, there was active concealment of the HIV status to avoid stigma from family, friends, and community. When disclosure occurred, there was selective disclosure to close family members who would keep the secret and respond favorably. Although the participants disclosed mostly to close family, some of their post-disclosure experiences included incidents of enacted stigma and discrimination. The fear of perceived stigma and self-stigma influenced the active concealment of their HIV status from others. Continuous concealment of one's HIV status and delayed disclosure limit the opportunities for support and care. There is a need to take into consideration the interaction between HIV-related stigma and disclosure to develop disclosure-counselling strategies in primary health care settings.


Pharmacokinetic profiles and egg residue patterns of levamisole in laying hens at two dosing rates and two routes of administration.
The levamisole maximum residue limit for edible fat, kidney, and muscle of chickens is 0.01 mg/kg. However, no maximum residue limit has been established for eggs. In the present study, the pharmacokinetic profile and levamisole residue in the eggs from laying hens were investigated using ultra-performance liquid chromatography-tandem mass spectrometry. A single dose of levamisole (30 mg/kg) was administered via the intramuscular or oral route, and an additional egg residue study was performed with 300 or 600 mg/kg commercial LEV drug (30 or 60 mg/kg as levamisole) orally. The limit of quantification was 0.0056 μg/mL and 0.0015 mg/kg for plasma and eggs, respectively. The plasma concentration was below the limit of quantification 10 and 12 h after intramuscular and oral administration, respectively. The half-life of the absorption phase was comparable between the intramuscular and oral routes, which was approximately 1 h, and the mean maximum concentration value was significantly higher in intramuscular (2.29 ± 0.30 μg/mL) than in oral (1.45 ± 0.38 μg/mL) route. The relative oral bioavailability after intramuscular administration was 92.3%. In the egg residue study, dose-dependent area under concentration and maximum concentration were observed after single oral administration of 30 and 60 mg/kg egg residue, and the calculated withdrawal period for both 30 and 60 mg/kg groups based on the positive list system standard (0.01 mg/kg) was 7 d after the treatment.


A Self-Reported Association between Temporomandibular Joint Disorders, Headaches, and Stress.
Temporomandibular joint disorder (TMD) is an umbrella term for a number of conditions in the area of the joint, temple, and masticatory system. Many of those with TMD also suffer from headaches and anxiety. The aims and objectives of this study were to determine if there exists an association between the Diagnostic Criteria of TMD (DC/TMD) symptom questionnaire and report of headaches as well as stress. A cross-sectional study was implemented via the use of the DC/TMD symptom questionnaire and the perceived stress scale (PSS). This was distributed conveniently among dental students in a multi-dental school setting in Riyadh, Saudi Arabia, and was completed by 152 dental students. Odds ratios, Chi-square, and their corresponding 95% confidence intervals are reported. Statistical significance was set at P ≤ 0.05. Those who ticked yes for pain on the TMD pain screener for pain with chewing hard or tough food, pain on mouth opening and lateral excursion, and pain with jaw habits as parafunction were at an odds risk from twice to thrice as likely to experience headaches. Similarly, those with poor coping skills on the PSS were more likely to have headaches (P = 0.002). Likewise, positive answers on the screener and symptom questionnaire were relevant with higher stress scores on the PSS. This study clarifies and reiterates the intertwined power of both stress and headaches; the former being a role player in TMD progression and the latter its product. TMD is unfortunately on the rise; it should not be brushed off as a nuisance. A multidisciplinary approach in diagnosis and treatment of both TMD and headaches by a team of orofacial pain specialist, neurologist, psychiatrist and/or psychologist, and physical therapists to untangle the deceiving presentation of both conditions would not only provide a more favorable prognosis but also improve cost and time expenditures.


[Oxygen balance and regional blood flow in the gingival mucosa during exposure to head-pelvis (+Gz) acceleration].
The information content of a new contact rheopolarographic procedure used to determine the oxygen balance and regional circulation of the gingival mucosa was measured, bearing in mind the applicability of the procedure as an objective index of human tolerance to +Gz acceleration. It was found that the parameters of the oxygen balance and regional circulation of the gingival mucosa were well correlated with blood pressure in the floor of the auricle. In contrast to the traditional methods for assessing tolerance to acceleration, the new procedure provides information about the health condition of the centrifuged subjects on a continuous basis. Variations in the oxygen balance and regional circulation of the gingival mucosa helped to identify compensatory reactions of the cardiovascular system in response to +Gz acceleration.


Pleural effusion following ovarian hyperstimulation.
Ovarian hyperstimulation syndrome (OHSS) is an iatrogenic complication that occurs in the luteal phase of an induced hormonal cycle. In most cases, the symptoms are self-limited and spontaneous regression occurs. However, severe cases are typically accompanied by acute respiratory distress. The objective of the present study was to describe the clinical presentation, treatment, and outcome of pleural effusion associated with OHSS in three patients undergoing in vitro fertilization. The patients ranged in age from 27 to 33 years. The onset of symptomatic pleural effusion (bilateral in all cases) occurred, on average, 43 days (range, 27-60 days) after initiation of hormone therapy for ovulation induction. All three patients required hospitalization for massive fluid resuscitation, and two required noninvasive mechanical ventilation. Although all three patients initially underwent thoracentesis, early recurrence of symptoms and pleural effusion prompted the use of drainage with a pigtail catheter. Despite the high output from the pleural drain (mean, 1,000 mL/day in the first week) and prolonged drainage (for 9-22 days), the outcomes were excellent: all three patients were discharged from hospital. Although pleural effusion secondary to OHSS is probably underdiagnosed, the associated morbidity should not be underestimated, especially because it affects potentially pregnant patients. In this study, early diagnosis and appropriate supportive measures yielded favorable results, limiting the surgical approach to adequate pleural drainage.


Rhabdanaerobium thermarum gen. nov., sp. nov., a novel anaerobic bacterium isolated from a hot spring.
A novel Gram-stain-positive, obligately anaerobic and thermophilic bacterium, strain SYSU GA15001T, was isolated from a sample collected from a hot spring located in Tengchong, China. Phylogenetic analysis based on the 16S rRNA gene sequence indicated that strain SYSU GA15001T belonged to the family Eubacteriaceae, and was closely related to 'Irregularibacter muris' 2PG-426-CC4.2T (93.6 % similarity), Garciella nitratireducens DSM 15102T (93.2 %) and Alkalibacter saccharofermentans Z-78920T (89.7 %). Cells were rods and motile by means of a subpolar flagellum. Spores were formed in old cultures. Optimum growth was observed at 45 °C, at pH 7 and in the presence of 1.5 % (w/v) NaCl. The strain utilized d-arabinose, dulcitol, d-galactose, glucose, lactose, maltose, d-mannose, raffinose, l-rhamnose, d-sorbitol, xylitol, l-alanine, l-arginine, glycine, l-isoleucine, l-leucine, dl-methionine, NH4Cl, l-proline and l-threonine as sole carbon/nitrogen sources. Additionally, complex substrates including beef extract, peptone, starch and yeast extract can also be used as sole energy sources. The major fatty acid methyl esters were C14 : 0, anteiso-C15 : 0 and C18 : 0. The polar lipid profile consisted of diphosphatidylglycerol, phosphatidylglycerol, phosphatidylinositol, an unidentified polar lipid, three unidentified phospholipids and an unidentified glycolipid. The DNA G+C content was determined to be 35.6 mol%. Strain SYSU GA15001T was physiologically and phylogenetically distinguishable from its closely related genera, and merits assignment as representing the type species of a new genus Rhabdanaerobium, with the name Rhabdanaerobium thermarum gen. nov., sp. nov. The type strain of the type species is SYSU GA15001T (=CGMCC 1.5219T=KCTC 15623T).


Bringing Needed Change to Medical Student Well-Being: A Call to Expand Accreditation Requirements.
Issue: Noting high rates of burnout, depression, and suicidality among medical students, academic medical communities are trying to identify preventive and curricular measures that protect and promote student well-being. To date, the effectiveness of these efforts is unclear. In addition, evidence increasingly suggests that the major drivers of distress appear to be factors within the social, learning, and work environments. Specific to medical schools in the United States, neither the Liaison Committee on Medical Education nor the Commission on Osteopathic College Accreditation include accreditation standards regarding well-being curricula and, as such, these curricula are not well-integrated into students' medical school experience. Current accreditation standards also do not specifically require institutions to assess or address systemic factors of the learning environment that negatively affect student well-being. Evidence: This paper proposes expanding current Liaison Committee on Medical Education and Commission on Osteopathic College Accreditation standards on professionalism to incorporate well-being as a core component of professional identity formation by requiring individual and institutional-level actions. Proposed changes to accreditation standards include (1) institutional assessment of the impact of the learning environment on student well-being; (2) continuous quality improvement efforts to address structural factors associated with student well-being and modification of practices that impair student well-being; and (3) integrated curriculum with related assessment to educate students on empirically-supported strategies for well-being. Implications: Refining undergraduate medical education accreditation standards in the United States to include language specific to student well-being will facilitate long overdue changes to the learning environment. In the end, the goal is not just to improve medical student well-being, but to provide a workforce better equipped for a sustainable and meaningful career.


Expression of a Schistosoma mansoni 28-kilodalton glutathione S-transferase in the livers of transgenic mice and its effect on parasite infection.
Schistosomiasis is a debilitating tropical disease for which an effective vaccine is needed. A 28-kDa glutathione S-transferase from Schistosoma mansoni (Sm28GST) has been shown to induce protective immunity. Sm28GST possesses significant sequence identity to mammalian GST isoforms. In order to study self-reactivity in mice immunized with Sm28GST and the concomitant phenomena of immune tolerance and epitope suppression, as well as their consequences for the protective immunity induced by this vaccination, we developed transgenic (Tg) mice that express Sm28GST under the control of a part of the mouse transferrin gene promoter. A study of (P28)Tg mice showed that the expression of Sm28GST was strictly localized in pericentrolobular hepatocytes. No histological change, inflammatory infiltrates, or modification of seric L-aspartate: 2-oxoglutarate aminotransferase concentration was observed over an 18-month period, despite a cross-reactivity between Sm28GST and a mouse molecule of 30 kDa. The immunoglobulin G anti-Sm28GST response of (P28)Tg mice immunized with recombinant Sm28GST was lower (P < 0.001) than that observed in non-(P28)Tg littermates and inversely proportional of Sm28GST liver expression. The response of non-(P28)Tg mouse spleen cells to Sm28GST stimulation was greater (P < 0.01) than that observed with (P28)Tg mouse spleen cells. (P28)Tg mice infected with 40 S. mansoni furcocercariae harbored more worms (P < 0.05) than did non-(P28)Tg control mice. The increase in the level of infection in (P28)Tg mice was reflected in concomitant increases in the numbers of adult worms and schistosome eggs found in livers and intestines after whole-body perfusion at 56 days postinfection, but no relative increase in the fertility of individual female worms was observed. The results obtained argue for the involvement of Sm28GST in reducing levels of infection and support the view that this enzyme has a central role in the maintenance of parasite viability, at least during its migration through host tissues.


Purification and characterization of the Saccharomyces cerevisiae RAD1/RAD10 endonuclease.
The Saccharomyces cerevisiae RAD1 and RAD10 genes are required for the incision step of excision repair, and in addition, they function in mitotic recombination. The RAD1 and RAD10 proteins are associated in a tight complex, and genetic studies have indicated that complex formation is essential for the RAD1/RAD10 controlled biological activities. We had previously purified the RAD10 protein to near homogeneity from yeast and shown that it is a DNA-binding protein with a strong preference for single-stranded DNA. In this study, we purify the RAD1 protein to near homogeneity from yeast and show that it also binds single-stranded DNA preferentially and that the RAD1/RAD10 complex possesses an endonuclease activity. We characterize the RAD1/RAD10 endonuclease activity on both single-stranded and double-stranded DNAs, using agarose gel electrophoresis and trichloroacetic acid precipitation. The RAD1/RAD10 nuclease exhibits a much higher level of activity on single-stranded DNA than double-stranded DNA. The susceptibility of double-stranded DNA to nicking by the RAD1/RAD10 enzyme is markedly dependent on the degree of negative superhelicity, such that a 15-fold increase in nicking rate is observed from superhelical state sigma = zero to sigma = -0.08. The enzyme produces 3'-hydroxyl and 5'-phosphate termini on both single- and double-stranded DNAs. We discuss the role of RAD1/RAD10 endonuclease in nucleotide excision repair and in mitotic recombination.


LRRC8 N termini influence pore properties and gating of volume-regulated anion channels (VRACs).
Volume-regulated anion channels (VRACs) are crucial for cell volume regulation and have various roles in physiology and pathology. VRACs were recently discovered to be formed by heteromers of leucine-rich repeat-containing 8 (LRRC8) proteins. However, the structural determinants of VRAC permeation and gating remain largely unknown. We show here that the short stretch preceding the first LRRC8 transmembrane domain determines VRAC conductance, ion permeability, and inactivation gating. Substituted-cysteine accessibility studies revealed that several of the first 15 LRRC8 residues are functionally important and exposed to a hydrophilic environment. Substituting glutamate 6 with cysteine decreased the amplitudes of swelling-activated ICl,vol currents, strongly increased iodide-over-chloride permeability, and markedly shifted the voltage dependence of channel inactivation. Importantly, these effects were reversed by 2-sulfonatoethyl methanethiosulfonate, which restores the negative charge at this amino acid position. Cd2+-mediated blocking of ICl,vol in cysteine variants suggested that the LRRC8 N termini come close together in the multimeric channel complex and might form part of the pore. We propose a model in which the N termini of the LRRC8 subunits line the cytoplasmic portion of the VRAC pore, possibly by folding back into the ion permeation pathway.