Utilization of a Directly Supervised Telehealth-Based Exercise Training Program in Patients With Nonalcoholic Steatohepatitis: Feasibility Study.
Most patients with nonalcoholic fatty liver disease (NAFLD) are physically inactive despite the well-known benefits of physical activity. Telehealth offers promise as a novel way to deliver an exercise training program and increase physical activity. However, the feasibility, safety, and efficacy of telehealth-based exercise programs is unknown in patients with NAFLD. The aim of this study was to determine the feasibility of a directly supervised exercise training program delivered exclusively with telehealth to patients with nonalcoholic steatohepatitis (NASH), the progressive form of NAFLD. In response to COVID-19 research restrictions, we adapted an existing clinical trial and delivered 20 weeks of moderate-intensity aerobic training 5 days a week under real-time direct supervision using an audio-visual telehealth platform. Aerobic training was completed by walking outdoors or using a home treadmill. Fitness activity trackers with heart rate monitors ensured exercise was completed at the prescribed intensity with real-time feedback from an exercise physiologist. Three female patients with biopsy-proven NASH were enrolled with a mean age of 52 (SD 14) years. The mean body mass index was 31.9 (SD 5.1) kg/m2. All patients had metabolic syndrome. All patients completed over 80% of exercise sessions (mean 84% [SD 3%]) and no adverse events occurred. Body weight (mean -5.1% [SD 3.7%]), body fat (mean -4.4% [SD 2.3%]), and waist circumference (mean -1.3 in. [SD 1.6 in.]) all improved with exercise. The mean relative reduction in magnetic resonance imaging-proton density fat fraction (MRI-PDFF) was 35.1% (SD 8.8%). Mean reductions in hemoglobin A1c and Homeostatic Model Assessment for Insulin Resistance were also observed (-0.5% [SD 0.2%] and -4.0 [SD 1.2], respectively). The mean peak oxygen consumption (VO2peak) improved by 9.9 (SD 6.6) mL/kg/min. This proof-of-concept study found that supervised exercise training delivered via telehealth is feasible and safe in patients with NASH. Telehealth-based exercise training also appears to be highly efficacious in patients with NASH, but this will need to be confirmed by future large-scale trials. ClinicalTrials.gov NCT03518294; https://clinicaltrials.gov/ct2/show/NCT03518294.


Assessment of 8-oxo-7,8-dihydro-2'-deoxyguanosine as a marker of oxidative DNA damage in gasoline filling station attendants.
The urinary excretion of 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodG) was used as a biomarker of oxidative DNA damage. The urinary 8-oxodG levels in petrol filling station attendants (exposed) at various petrol bunks were estimated as well as in the unexposed (cashier) population. A total of 100 workers (79 petrol fillers and 21 cashiers) aged from 20 to 41 years participated in the study. An informed consent was taken from each participant. Information on personal habits and health was obtained through a questionnaire. After shifts, urine samples were collected analyzed for 8-oxodG using enzyme-linked immunosorbent assay (ELISA). Fifty-three percent of workers were in the 21-30 years age group. The maximum level of 8-oxodG was observed in the age group ≥ 41 years and the minimum in the age group of 31-40 years. The maximum level of 8-oxodG was observed among those workers who had ≥ 21 years of experience. The concentrations of 8-oxodG were significantly higher in petrol fillers than those in cashiers (p < 0.05). Despite the conflicting results obtained in our study it was shown that 8-oxodG is related to chemical exposure. Further research is needed embracing a bigger number of participants to highlight the correlations between the exposure and the effects.


Ligand field theory and the origin of life as an emergent feature of the periodic table of elements.
The assumption that all biological catalysts are either proteins or ribozymes leads to an outstanding enigma of biogenesis-how to determine the synthetic pathways to the monomers for the efficient formation of catalytic macromolecules in the absence of any such macromolecules. The last 60 years have witnessed chemists developing an understanding of organocatalysis and ligand field theory, both of which give demonstrable low-molecular-weight catalysts. We assume that transition-metal-ligand complexes are likely to have occurred in the deep ocean trenches by the combination of naturally occurring oceanic metals and ligands synthesized from the emergent CO(2), H(2), NH(3), H(2)S, and H(3)PO(4). We are now in a position to investigate experimentally the metal-ligand complexes, their catalytic function, and the reaction networks that could have played a role in the development of metabolism and life itself.


Congenital hydrocephalus and ventriculoperitoneal shunts: influence of etiology and programmable shunts on revisions.
Hydrocephalus is a notorious neurosurgical disease that carries the adage "once a shunt always a shunt." This study was conducted to review the treatment results of pediatric hydrocephalus. Pediatric patients who underwent ventriculoperitoneal shunt surgery over the past 14 years were reviewed for shunt revisions. Variables studied included age at shunt placement, revision, or replacement; programmable shunts; infection; obstruction; and diagnosis (congenital, posthemorrhagic, craniospinal dysraphism, and others including trauma, tumors, and infection). Multiple regression analysis methods were used to determine independent risk factors for shunt failure and the number of shunt revisions. The Kaplan-Meier method of survival analysis was used to compare etiologies on the 5-year survival (revision-free) rate and the median 5-year survival time. A total of 253 patients were studied with an almost equal sex distribution. There were 92 patients with congenital hydrocephalus, 69 with posthemorrhagic hydrocephalus, 48 with craniospinal dysraphism, and 44 with other causes. Programmable shunts were used in 73 patients (other types of shunts were used in 180 patients). A total of 197 patients (78%) underwent revision surgeries due to shunt failures. The mortality rate was 1.6%. Age at first revision, the 5-year survival rate, and the median 5-year survival time were significantly less for both posthemorrhagic and craniospinal dysraphism than for either the congenital or "other" group (p < 0.05). The failure rate and number of revisions were not significantly reduced with programmable shunts compared with either pressure-controlled or no-valve shunts (p > 0.5). Posthemorrhagic hydrocephalus and craniospinal dysraphism hydrocephalus had significantly earlier revisions than congenital and other etiologies. Programmable systems did not reduce the failure rate or the average number of shunts revisions.


Comparative Transcriptomics of the Bovine Apicomplexan Parasite Theileria parva Developmental Stages Reveals Massive Gene Expression Variation and Potential Vaccine Antigens.
Theileria parva is a protozoan parasite that causes East Coast fever (ECF), an economically important disease of cattle in Africa. It is transmitted mainly by the tick Rhipicephalus appendiculatus. Research efforts to develop a subunit vaccine based on parasite neutralizing antibodies and cytotoxic T-lymphocytes have met with limited success. The molecular mechanisms underlying T. parva life cycle stages in the tick vector and bovine host are poorly understood, thus limiting progress toward an effective and efficient control of ECF. Transcriptomics has been used to identify candidate vaccine antigens or markers associated with virulence and disease pathology. Therefore, characterization of gene expression throughout the parasite's life cycle should shed light on host-pathogen interactions in ECF and identify genes underlying differences in parasite stages as well as potential, novel therapeutic targets. Recently, the first gene expression profiling of T. parva was conducted for the sporoblast, sporozoite, and schizont stages. The sporozoite is infective to cattle, whereas the schizont is the major pathogenic form of the parasite. The schizont can differentiate into piroplasm, which is infective to the tick vector. The present study was designed to extend the T. parva gene expression profiling to the piroplasm stage with reference to the schizont. Pairwise comparison revealed that 3,279 of a possible 4,084 protein coding genes were differentially expressed, with 1,623 (49%) genes upregulated and 1,656 (51%) downregulated in the piroplasm relative to the schizont. In addition, over 200 genes were stage-specific. In general, there were more molecular functions, biological processes, subcellular localizations, and pathways significantly enriched in the piroplasm than in the schizont. Using known antigens as benchmarks, we identified several new potential vaccine antigens, including TP04_0076 and TP04_0640, which were highly immunogenic in naturally T. parva-infected cattle. All the candidate vaccine antigens identified have yet to be investigated for their capacity to induce protective immune response against ECF.


Dynamic contrast-enhanced MRI findings of acute pancreatitis in ectopic pancreatic tissue: case report and review of the literature.
Acute pancreatitis in ectopic pancreatic tissue is an uncommon cause of acute abdominal pain and can be difficult to diagnose on imaging. Our aim is to raise awareness and aid in the diagnosis of this entity by highlighting helpful dynamic contrast-enhanced MRI imaging findings. We report a 51-year-old man with acute onset epigastric pain presented to ER. With the presence of elevated serum lipase, the clinical diagnosis of acute pancreatitis was made. Contrast enhanced CT demonstrated normal pancreas and a focal mass at the duodenojejunal flexure, mimicked a neoplasm. Subsequent dynamic contrast enhanced MR images demonstrated enhancement pattern of the lesion similar to the native pancreatic tissue enhancement, a finding raised the possibility of acute pancreatitis in ectopic pancreatic tissue, but tumor was not excluded. Finally, patient undergone surgical bowel resection including the suspected mass that was proved as an ectopic pancreatic tissue on microscopic examination. We concluded that findings on dynamic contrast enhanced MR imaging can be characteristic and diagnostic of acute pancreatitis in ectopic pancreatic tissue in the appropriate clinical setting.


Development of a practical animal model of photodynamic therapy using a high concentration of extracellular talaporfin sodium in interstitial fluid: influence of albumin animal species on myocardial cell photocytotoxicity in vitro.
Photodynamic reaction-induced photocytotoxicity using talaporfin sodium is inhibited by serum proteins binding to talaporfin sodium. The serum albumin binding site for talaporfin sodium differs among animal species. To identify a practical animal therapeutic model, we studied the ability of human, canine, bovine, and porcine albumin to influence talaporfin sodium-induced photocytotoxicity in rat myocardial cells in vitro. Human, canine, bovine, and porcine serum albumins were used. The ratio of talaporfin sodium binding, which is strongly associated with photocytotoxicity, was measured by ultrafiltration with an albumin concentration of 0.5-20 mg/ml and 20 μg/ml talaporfin sodium to mimic interstitial fluid. Rat myocardial cell lethality was measured by the WST assay 2 h after samples were exposed to a radiant exposure of 20 J/cm2 by a red diode laser (Optical Fuel™, Sony, Tokyo, Japan) with a wavelength of 663 nm. The binding ratio dependence on albumin concentration differed among the animal species. Bovine albumin exhibited the largest difference from human albumin, with a maximum difference of 31% at 2 mg/ml albumin. The cell lethality characteristic was similar between human and canine albumin. The cell lethality dependence on albumin was not in the same order as the binding ratio. Cell lethality was lowest for human albumin with higher albumin concentrations between 5 and 20 mg/ml. There were no significant differences in cell lethality between bovine and porcine albumin and between human and canine albumin. We suggest that the canine model may be a useful animal therapeutic model for evaluating photodynamic therapy using a high concentration of the photosensitizer in the extracellular space.


Role of manometry, defecography and anal endosonography in the evaluation of colorectal disorders.
altered motility or anatomy of the rectum, anus and perineal floor may lead to symptoms which are unresponsive to routine therapeutic approaches. These disturbances usually lead to constipation, fecal incontinence, or both. Different tests and techniques for evaluating anorectal and perineal disorders, developed in the last two decades, make a better understanding of these disorders possible. This study was designed to evaluate the diagnostic benefits of combining manometry, defecography and anal endosonography in the assessment of patients with anorectal disorders. twenty-five children with constipation (with or without soiling), incontinence and/or prolapse underwent anal manometry, defecography and anal endosonography. Group A consisted of 9 children with fecal incontinence, group B consisted of 10 children with constipation with soiling, and group C comprised 6 children with constipation without soiling. in group A resting incontinence was associated with a hypotonic external sphincter in 4 out of 9 patients, 2 of whom had internal anal sphincter thinning. In group B resting incontinence was associated with a hypotonic external sphincter in 8 out of 10 patients, 6 of whom had internal anal sphincter thinning. In group C these associations were not seen in any of the patients. barium enema is not sufficient for an accurate diagnosis of anorectal disorders. No single test is capable of revealing the type of disease. Anal manometry, defecography and endosonography are complementary procedures in the assessment of this group of disorders. This new approach will improve our knowledge of the pathogenesis of these disorders in children. However, further studies are needed to obtain conclusive evidence.


Effective cross sections for production of single-strand breaks in plasmid DNA by 0.1 to 4.7 eV electrons.
We determined effective cross sections for production of single-strand breaks (SSBs) in plasmid DNA [pGEM 3Zf(-)] by electrons of 10 eV and energies between 0.1 and 4.7 eV. After purification and lyophilization on a chemically clean tantalum foil, dry plasmid DNA samples were transferred into a high-vacuum chamber and bombarded by a monoenergetic electron beam. The amount of the circular relaxed DNA in the samples was separated from undamaged molecules and quantified using agarose gel electrophoresis. The effective cross sections were derived from the slope of the yield as a function of exposure and had values in the range of 10(-15)- 10(-14) cm2, giving an effective cross section of the order of 10(-18) cm2 per nucleotide. Their strong variation with incident electron energy and the resonant enhancement at 1 eV suggest that considerable damage is inflicted by very low-energy electrons to DNA, and it indicates the important role of pi* shape resonances in the bond-breaking process. Furthermore, the fact that the energy threshold for SSB production is practically zero implies that the sensitivity of DNA to electron impact is universal and is not limited to any particular energy range.


New species and new record of Megatyrus Hlaváč & Nomura (Coleoptera: Staphylinidae: Pselaphinae) from Asia.
A new species, Megatyrus femoralis sp. n., is described from the Koshi Zone, East Nepal, with major diagnostic features illustrated. Megatyrus masumotoi Nomura, Sakchoowong & Chanpaisaeng, originally described from southwestern Thailand, is recorded from the Noring Timur Mountain, West Malaysia. The above data extends the known range of Megatyrus about 1,200 km to the west, and 870 km to the south.


Arsenic concentrations in local aromatic and high-yielding hybrid rice cultivars and the potential health risk: a study in an arsenic hotspot.
The presence of high levels of arsenic (As) in rice fields has negative effects on the health of those consuming rice as their subsistence food. This study determined the variation in total As concentration in local aromatic rice (LAR) (kalijira) and two high-yielding varieties (HYVs) (BRRI dhan 32 and BRRI dhan 28) grown in paddy fields in Matlab, Bangladesh, an As hotspot with elevated As levels in groundwater. Mature rice grain samples and soil samples were collected from different paddy fields, and the As concentrations in both the de-husked grains and the husks of the three rice cultivars were analysed to identify the safest of the three cultivars for human consumption. The results showed that the total As concentration was higher (0.09-0.21 mg As kg-1) in the de-husked grains of LAR than in the husks, while the opposite was found for the HYV rice. Moreover, the As concentration in soil samples was 2 to 5-fold higher for the LAR than for the HYVs, but the As accumulation factor (AF) was lower in the LAR (0.2-0.4%) than in the HYVs (0.9-1%). Thus, LAR can be considered the safest of the three cultivars for human consumption owing to its low AF value. Furthermore, due to the low AF, growing LAR instead of HYVs in soils with slightly elevated As levels could help improve the food safety level in the food chain.


Children's Responses to Maltreatment: Key Conclusions From a Systematic Literature Review.
Child maltreatment (CM) has received much research attention in recent years, leading to the enormous development of services provided to children worldwide. However, although the literature in the field of trauma studies recognizes that responses to trauma are of central importance, for a variety of reasons, the research on this subject is sparse. The current systematic literature review was therefore designed to characterize and analyze the existing knowledge on children's responses to CM, while putting in focus all possible responses that will appear in the identified manuscripts. Key databases were explored using the Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines, and only six studies were identified as relevant to the focus of the current study. Utilizing thematic analysis, the results addressed four main themes: the studies' methodologies, the terminology and responses that were explored in these studies, the abuse-related factors and contexts that were taken into account, and the consequences for the children (of their responses to the maltreatment). A key conclusion from this review is that concepts in this area, as well as theory, are considerably lacking. Promoting the development of theory regarding children's responses to CM is urgently needed, as it will greatly impact clinical intervention, legal investigation, and the development of prevention programs.


Stereoselectivity of macrocyclic ring-closing olefin metathesis.
[reaction: see text] Macrocyclic ring-closing olefin metathesis using ruthenium catalyst 3 was performed to produce a 14-membered lactone. The E/Z ratio of lactone was high regardless of the R group (auxiliary) or the initial alkene stereochemistry. A kinetic study demonstrates that the high E/Z ratio is due to secondary metathesis reactions that isomerize the product to the thermodynamic E/Z ratio.


Changes in the Phenotype of Intramural Inhibitory Neurons of the Porcine Descending Colon Resulting from Glyphosate Administration.
Environmental contamination and the resulting food contamination represent a serious problem and pose a major threat to animal and human health. The gastrointestinal tract is directly exposed to a variety of substances. One is glyphosate, whose presence in the soil is commonly observed. This study demonstrates the effects of low and high glyphosate doses on the populations of intramural neurons of the porcine descending colon. An analysis was performed on neurons ex-pressing the vasoactive intestinal peptide, pituitary adenylate cyclase-activating peptide, a neuronal isoform of nitrogen oxide synthase, and galanin. Even a low dose of glyphosate increased the number of neurons immunoreactive against the studied substances. However, the changes depended on both the plexus analysed and the substance tested. Meanwhile, a high glyphosate dose resulted in quantitative changes (an increase in the number) within neurons immunoreactive against all the studied neuropeptides/enzymes in the myenteric plexus and both submucosal plexuses. The response of the enteric nervous system in the form of an increase in the number of neurons immunoreactive against neuroprotective substances may suggest that glyphosate has a toxic effect on enteric neurons which attempt to increase their survivability through the released neuroprotective substances.


Characteristics of Microbial Factors of Healthcare-Associated Infections Including Multidrug-Resistant Pathogens and Antibiotic Consumption at the University Intensive Care Unit in Poland in the Years 2011-2018.
Introduction: In recent years, an increase in healthcare-associated infections (HAIs) caused by resistant pathogens, which is a clinically troublesome trend, has been observed. The aim of the study was to analyze the microbial factors of HAIs and the drug resistance of microorganisms to selected antibiotics and their consumption. Material and Methods: The retrospective study included 3708 patients hospitalized in the intensive care unit (ICU) of the University Teaching Hospital in Wroclaw, who were diagnosed with 742 HAIs in the period from 1 January 2011 to 31 December 2018. The aim of the study was the analysis of microorganisms isolated in the respective clinical forms of HAIs, including the occurrence of "alert pathogens", presence of multidrug-resistant (MDR) strains, and consumption of selected antibiotics. Findings: During the study period, 846 microorganisms were cultured in patients with HAIs, and among them, Acinetobacter baumannii MDR represented 31.8%; Klebsiella pneumoniae ESBLs, 11.3%; Pseudomonas aeruginosa MDR, 4.1% and MRSA, 2.2%; and Enterococcus spp. vancomycin-resistant enterococci (VRE), 1.3%. Among all the pathogens, Gram-negative bacteria (GNB) were dominant (71.6%). Gram-positive bacteria and fungi accounted for 21.6% and 7%, respectively. The total number of strains responsible for ventilator-associated pneumonia (VAP), urinary tract infection (UTI), and central line-associated blood stream infection (CLA-BSI) was as follows: 458 (54.1%), 274 (32.4%), and 114 (13.5%), respectively. Among the etiological factors of VAP, there was a prevalence of A. baumannii MDR (41.9%), as well as in the case of UTI (21.9%). With regards to CLA-BSI, MRCNS (29.8%) was the dominant pathogen. The "alert pathogens" accounted for 54.7% of all the analyzed strains. The MDR strains represented 72.6% and 9.7% among A. baumannii and P. aeruginosa, respectively. In the years 2011 vs. 2018, an increase in infections with MDR bacilli was observed, 34.6% vs. 61.0% (p = 0.0008), respectively, including A. baumannii MDR 16.54% vs. 34.56 % (p = 0.0009) and Enterobacterales ESBL+/AMPC 11.8% vs. 15.44 % (p = 0.3921). Resistance to methicillin was confirmed in 35.2% of S. aureus strains. Resistance to vancomycin was found among 30.9% of Enterococcus spp. The observed period was marked by an increase in the consumption of carbapenems: 197.7 vs. 235.9 defined daily dose (DDD)/1000 patients-days. Conclusions: Gram-negative bacteria were found to be dominant pathogens in healthcare-associated infections. The most frequently cultured pathogens were multidrug-resistant A. baumannii, K. pneumoniae ESBL(+), and P. aeruginosa. The study showed an increase in the incidence of "alert pathogens" and MDR bacilli, as well as the tendency of a growing resistance to antibiotics during the observed period. Microbiological analysis of HAIs and the consumption of antibiotics is the necessary element of the proper antibiotic policy in hospitals.


Incidence trends of childhood type 1 diabetes in eastern Saudi Arabia.
To assess the epidemiology and incidence rate of type 1 diabetes in children <15 years of age in a subpopulation in the eastern province of the Kingdom of Saudi Arabia (KSA). This is a subpopulation-based observational incidence study. Admission dates, diagnosis, age, and gender were collected for all Saudi children <15 years of age with new onset type 1 diabetes that received medical care at the Saudi Aramco Medical Services Organization, Dhahran Health Center, Dhahran, KSA during the period 1990-2007. The case ascertainment rate was estimated to be 100%. There were 438 patients diagnosed during the study period. The average incidence rate was 27.52/100,000/year (95% CI: 26.72-28.32), increasing from 18.05/100,000/year in the first 9 years of the study period to 36.99/100,000/year in the last 9 years. Twenty-one percent of patients were <5 years of age, with no significant difference in the rate of increase in the incidence rate of this age group compared to the older age groups. Of the total patients, 55% were females, and 40% presented with diabetic ketoacidosis. The incidence rate of childhood type 1 diabetes increased alarmingly over the past 18 years in our study population. A Kingdom-wide diabetes registry is essential to study the epidemiology of this disease in the whole country.


Visual Acuity Outcomes after Cataract Extraction with Intraocular Lens Implantation in Eyes with Diabetic Retinopathy.
To evaluate change in best-corrected visual acuity (BCVA) in patients with diabetes and diabetic retinopathy (DR) after cataract extraction (CE). Retrospective cohort study. Diabetic eyes of patients 18 years of age or older that underwent CE at the Cleveland Clinic from 2012 through 2018. Chart review examining visual acuities from patient visits before and after surgery, as well as OCT images. Statistical analysis was carried out using multiple linear regression models. The primary end point was change in BCVA during the first postoperative year. The secondary end point was the association of central subfield thickness at baseline with change in BCVA. Additional preoperative factors examined were age, race, gender, laterality, insulin use, hemoglobin A1c, creatinine, blood urea nitrogen, and estimated glomerular filtration rate. Diabetic eyes without DR (n = 138) and eyes with mild/moderate nonproliferative DR (NPDR; n = 125), severe NPDR (n = 20), and proliferative DR (PDR; n = 72) were included. A year after surgery, eyes without DR gained a median of 11.0 Early Treatment Diabetic Retinopathy Study (ETDRS) letters (interquartile range [IQR], 5.0-20.0) from 65.0 (IQR, 58.0-70.0) before surgery, eyes with mild or moderate DR gained 10.0 letters (IQR, 5.0-22.0) from 65.0 (IQR, 58.0-76.0), eyes with severe NPDR gained 20.5 letters (IQR, 8.0-28.5) from 55.0 (IQR, 26.0-65.0), and eyes with PDR gained 15.0 letters (IQR, 6.0-29.5) from 55.0 (IQR, 35.0-61.0). Eyes without DR or mild or moderate NPDR showed significantly greater improvements in VA 1 year after surgery compared with eyes with severe NPDR or PDR when controlling for baseline visual acuity (VA), with eyes with more severe DR showing less expected VA gain (P < 0.001). Length of disease most strongly related to baseline DR severity (ρ = 0.431; P < 0.001). Cataract surgery is beneficial in most patients with DR without severe concurrent macular pathologic features. However, preoperative VA and the severity of DR can limit visual outcomes.


How is mortality affected by money, marriage, and stress?
It is believed that the length of a person's life depends on a mixture of economic and social factors. Yet the relative importance of these is still debated. We provide recent British evidence that marriage has a strong positive effect on longevity. Economics matters less. After controlling for health at the start of the 1990s, we cannot find reliable evidence that income affects the probability of death in the subsequent decade. Although marriage keeps people alive, it does not appear to work through a reduction of stress levels. Greater levels of psychological distress (as measured by General Health Questionnaire (GHQ) stress scores) cannot explain why unmarried people die younger. For women, however, we do find that mental strain itself is dangerous. High GHQ stress scores help to predict the probability of an early death.


Prosthodontic complications of Spline dental implants.
Access to accurate clinical data on the frequency of prosthodontic complications encountered in dental implant restorations can provide dentists with important information for evaluating dental implant systems. The complication of prosthetic screw loosening, for example, has often been ignored or under-reported. Only limited published clinical data exists on the use of the Spline connections for dental implant restorations. This paper reports on an in-vitro investigation of prosthodontic complications encountered with the Spline dental implant system restored by students and residents at The Ohio State University's School of Dentistry. From a comprehensive list of implant patients generated from the University's computer records, an extensive chart review was conducted, and all patients who were treated with Spline dental implants were entered into the study. During a second chart review, data on all prosthodontic complications with Spline implant restorations were gathered on reporting forms then entered into a computer database and subjected to statistical analysis. The results showed that 95.75% of the prostheses were free of any complications. Iatrogenic factors that resulted in screw loosening were identified. Based on the findings of this relatively short-term study ranging from 29 to 59 months of post-restoration clinical follow-up, Spline dental implants provided a stable implant prosthetic connection.


Importance of the penultimate positive charge in mouse hepatitis coronavirus A59 membrane protein.
The coronavirus membrane (M) protein carboxy tail interacts with the nucleocapsid during virus assembly. Previous studies demonstrated that the two terminal residues are important, and the charged residue (R227) in the penultimate position in the mouse hepatitis coronavirus (MHV) A59 M protein was suggested to participate in intermolecular interactions with negative charges in the nucleocapsid (N) protein. To determine the significance of the positive charge at position 227, we substituted the arginine with lysine (K), aspartic acid (D), glutamic acid (E), or alanine (A) and studied these by reverse genetics in the context of a MHV full-length infectious clone. Viruses with wild-type phenotype were readily recovered with the K or A substitutions. In contrast, negative-charge substitutions were not tolerated as well. In all recovered R227D viruses the negative charge was replaced with heterologous residues resulting from apparent template switching during negative-strand synthesis of subgenomic RNA 7. An additional second-site compensatory V202I substitution was present in some viruses. Recovered R227E viruses had second-site changes within the M protein carboxy tail that were partially compensatory. Significantly, most of the second site changes in the R227E mutant viruses were previously shown to compensate for the removal of negative charges in the N protein. Our results strongly indicate that a positive charge is not absolutely required. It is clear that other regions within the tail must also be involved in helping mediate interactions between the M protein and the nucleocapsid.


Mesocestoides corti (syn. M. vogae): modulation of larval motility by neuropeptides, serotonin and acetylcholine.
The effect of platyhelminth FaRPs and selected classical neurotransmitters on the motility of intact Mesocestoides corti (syn. M. vogae) tetrathyridial larvae was studied in vitro using a micromotility meter. The effects of the test substances were temperature dependent and these were examined at 4, 23, 30 and 36 degrees C. At 36 degrees C all test substances had concentration-dependent excitatory effects, with thresholds for activity of: 100 nM (GNFFRFamide), 10 microm (YIRFamide), 30 microM (GYIRFamide), 100 nM (serotonin) and 100 microM (acetylcholine). At this temperature significant elevation of motility indices (MI) was recorded within 5 min of the addition of peptide or serotonin. The effect of acetylcholine was slower in onset and appeared 15-20 min post-addition. At 30 degrees C larval motility diminished more rapidly than that recorded at 36 degrees C, following the addition of 1 mM of each test substance. At 23 degrees C only serotonin (1 mM) significantly increased the MI, all other test substances having no apparent effect. Larval movement was completely arrested at 4 degrees C. The results demonstrate for the first time excitatory effects of platyhelminth neuropeptides and acetylcholine on muscle systems in cestode larvae. The fact that the only known cestode FaRP, GNFFRF amide, was more potent than any of the turbellarian FaRPs tested, suggests structural conservation of FaRPs and FaRP receptors within the cestodes.


Multicenter evaluation of the Elecsys Toxo IgG and IgM tests for the diagnosis of infection with Toxoplasma gondii.
Detection of IgG and IgM antibodies is commonly performed for the diagnosis of infection with Toxoplasma gondii. We determined the accuracy of the Elecsys Toxo IgG and IgM test at four European laboratories compared to local reference methods. Coefficients of variation for reproducibility ranged from 1.0 to 6.5% for IgG and from 0.8 to 3.2% for IgM. Seroconversion panels revealed high overall concordance with the reference tests. The Elecsys test detected IgG antibodies earlier than the Cobas Core IgG test in 19 of 47 panels; persisting IgM antibodies were observed in the VIDAS but not the Elecsys test in five of 47 panels. In 31.4% of latent stage sera with persistent IgM antibodies (positive LIASON IgM), the Elecsys IgM test gave negative results indicating increased "clinical" specificity. Sensitivity and specificity of the Elecsys IgG assay ranged from 99.45 to 100% and 87.50-99.80%, respectively, and 91.11-95.74 and 98.45-99.79% for the Elecsys IgM assay, respectively. In conclusion, excellent reproducibility and accuracy make the Elecsys Toxo G and M tests highly suitable for the detection of anti-T. gondii IgG and IgM antibodies. The lower detection rates for persistent IgM in the Elecsys IgM test increase "clinical" specificity and decrease the need for follow-up testing.


Eye Movement Abnormalities in Major Depressive Disorder.
Background: Despite their high lifetime prevalence, major depressive disorder (MDD) is often difficult to diagnose, and there is a need for useful biomarkers for the diagnosis of MDD. Eye movements are considered a non-invasive potential biomarker for the diagnosis of psychiatric disorders such as schizophrenia. However, eye movement deficits in MDD remain unclear. Thus, we evaluated detailed eye movement measurements to validate its usefulness as a biomarker in MDD. Methods: Eye movements were recorded from 37 patients with MDD and 400 healthy controls (HCs) using the same system at five University hospitals. We administered free-viewing, fixation stability, and smooth pursuit tests, and obtained 35 eye movement measurements. We performed analyses of covariance with group as an independent variable and age as a covariate. In 4 out of 35 measurements with significant group-by-age interactions, we evaluated aging effects. Discriminant analysis and receiver operating characteristic (ROC) analysis were conducted. Results: In the free-viewing test, scanpath length was significantly shorter in MDD (p = 4.2 × 10-3). In the smooth pursuit test, duration of saccades was significantly shorter and peak saccade velocity was significantly lower in MDD (p = 3.7 × 10-3, p = 3.9 × 10-3, respectively). In the fixation stability test, there were no significant group differences. There were significant group differences in the older cohort, but not in the younger cohort, for the number of fixations, duration of fixation, number of saccades, and fixation density in the free-viewing test. A discriminant analysis using scanpath length in the free-viewing test and peak saccade velocity in the smooth pursuit showed MDD could be distinguished from HCs with 72.1% accuracy. In the ROC analysis, the area under the curve was 0.76 (standard error = 0.05, p = 1.2 × 10-7, 95% confidence interval = 0.67-0.85). Conclusion: These results suggest that detailed eye movement tests can assist in differentiating MDD from HCs, especially in older subjects.


Post-transplant malignancies in alcoholic liver disease.
Post-transplant malignancy is emerging as an important cause of mortality in patients with cirrhosis undergoing liver transplant (LT). However, establishing the exact relationship between the two needs further evaluation. It has been observed that approximately 30% deaths after 10 years of hepatic transplantation occur due to de novo malignancies. Various known risk factors include immunosuppression, age of patient, alcoholic liver disease (ALD) or primary sclerosing cholangitis, smoking, and oncogenic viral infections. There is scanty literature on the post-transplant malignancy risk in patients with alcoholic cirrhosis. The current evidence suggests a particularly increased risk of oropharyngeal and lung cancers in patients transplanted for ALD. Abstinence from alcohol, smoking and other tobacco-containing products along with optimization of immunosuppression are paramount for decreasing the risk of post-transplant malignancies.


Automated design of 3D DNA origami with non-rasterized 2D curvature.
Improving the precision and function of encapsulating three-dimensional (3D) DNA nanostructures via curved geometries could have transformative impacts on areas such as molecular transport, drug delivery, and nanofabrication. However, the addition of non-rasterized curvature escalates design complexity without algorithmic regularity, and these challenges have limited the ad hoc development and usage of previously unknown shapes. In this work, we develop and automate the application of a set of previously unknown design principles that now includes a multilayer design for closed and curved DNA nanostructures to resolve past obstacles in shape selection, yield, mechanical rigidity, and accessibility. We design, analyze, and experimentally demonstrate a set of diverse 3D curved nanoarchitectures, showing planar asymmetry and examining partial multilayer designs. Our automated design tool implements a combined algorithmic and numerical approximation strategy for scaffold routing and crossover placement, which may enable wider applications of general DNA nanostructure design for nonregular or oblique shapes.


Oral immunization with the saliva-binding region of Streptococcus mutans AgI/II genetically coupled to the cholera toxin B subunit elicits T-helper-cell responses in gut-associated lymphoid tissues.
Mice immunized intragastrically (i.g.) with a genetically constructed chimeric protein consisting of the saliva-binding region (SBR) of Streptococcus mutans AgI/II coupled to cholera toxin (CT) A2 and B subunits (CTA2/B) develop serum immunoglobulin G (IgG) and mucosal IgA antibody responses against AgI/II that are enhanced by the coadministration of CT as an adjuvant. To investigate the development of antigen-specific T cells in the gut-associated lymphoid tissues, mice were immunized i.g. with SBR, SBR-CTA2/B, or SBR-CTA2/B plus CT. AgI/II-specific T cells in Peyer's patches (PP), mesenteric lymph nodes (MLN), and spleen were assayed by lymphoproliferation and flow cytometry for the expression of T-cell surface markers, and cytokine mRNA expression was evaluated by reverse transcription-PCR. T-cell responses were consistent with antibody responses but were detectable after the first immunization. Proliferative responses of PP and MLN cells upon stimulation with AgI/II in vitro were low and delayed in mice given SBR alone, and these cells displayed a mixed type 1 and 2 (or Th0) pattern of cytokine expression. Immunization with SBR-CTA2/B resulted in greater AgI/II-specific proliferative responses in PP cells and an increase in the proportion of CD4+ T cells. Coadministration of CT with SBR-CTA2/B led to greater proliferative responses especially in the MLN cells, which then showed an increase in CD4+ cells. Immunization with SBR-CTA2/B (with or without CT) skewed the cytokine expression pattern in PP and MLN cells toward Th2. The results indicate that T helper cells were induced in gut-associated lymphoid tissues by i.g. immunization with SBR-CTA2/B, concomitantly with and prior to the appearance of circulating and mucosal antibodies.


Prognostic implications of asymptomatic left ventricular dysfunction in patients undergoing vascular surgery.
The prognostic value of heart failure symptoms on postoperative outcome is well acknowledged in perioperative guidelines. The prognostic value of asymptomatic left ventricular (LV) dysfunction remains unknown. This study evaluated the prognostic implications of asymptomatic LV dysfunction in vascular surgery patients assessed with routine echocardiography. Echocardiography was performed preoperatively in 1,005 consecutive vascular surgery patients. Systolic LV dysfunction was defined as LV ejection fraction less than 50%. Ratio of mitral-peak velocity during early and late filling, pulmonary vein flow, and deceleration time was used to diagnose diastolic LV dysfunction. Troponin-T measurements and electrocardiograms were performed routinely perioperatively. Multivariate regression analyses evaluated the relation between LV function and the study endpoints, 30-day cardiovascular events, and long-term cardiovascular mortality. Left ventricular dysfunction was diagnosed in 506 (50%) patients of which 80% were asymptomatic. In open vascular surgery (n = 649), both asymptomatic systolic and isolated diastolic LV dysfunctions were associated with 30-day cardiovascular events (odds ratios 2.3, 95% confidence interval [CI] 1.4-3.6 and 1.8, 95% CI 1.1-2.9, respectively) and long-term cardiovascular mortality (hazard ratios 4.6, 95% CI 2.4-8.5 and 3.0, 95% CI 1.5-6.0, respectively). In endovascular surgery (n = 356), only symptomatic heart failure was associated with 30-day cardiovascular events (odds ratio 1.8, 95% CI 1.1-2.9) and long-term cardiovascular mortality (hazard ratio 10.3, 95% CI 5.4-19.3). This study demonstrated that asymptomatic LV dysfunction is predictive for 30-day and long-term cardiovascular outcome in open vascular surgery patients. These data suggest that preoperative risk stratification should include not only solely heart failure symptoms but also routine preoperative echocardiography to risk stratify open vascular surgery patients.


A High-Performance Antibacterial Nanostructured ZnO Microfluidic Device for Controlled Bacterial Lysis and DNA Release.
In this work, the antibacterial properties of nanostructured zinc oxide (ZnO) surfaces are explored by incorporating them as walls in a simple-to-fabricate microchannel device. Bacterial cell lysis is demonstrated and quantified in such a device, which functions due to the action of its nanostructured ZnO surfaces in contact with the working fluid. To shed light on the mechanism responsible for lysis, E. coli bacteria were incubated in zinc and nanostructured ZnO substrates, as well as the here-investigated ZnO-based microfluidic devices. The unprecedented killing efficiency of E. coli in nanostructured ZnO microchannels, effective after a 15 min incubation, paves the way for the implementation of such microfluidic chips in the disinfection of bacteria-containing solutions. In addition, the DNA release was confirmed by off-chip PCR and UV absorption measurements. The results indicate that the present nanostructured ZnO-based microfluidic chip can, under light, achieve partial inactivation of the released bacterial DNA via reactive oxygen species-mediated oxidative damage. The present device concept can find broader applications in cases where the presence of DNA in a sample is not desirable. Furthermore, the present microchannel device enables, in the dark, efficient release of bacterial DNA for downstream genomic DNA analysis. The demonstrated potential of this antibacterial device for tailored dual functionality in light/dark conditions is the main novel contribution of the present work.


Do ethical guidelines give guidance? A critical examination of eight ethics regulations.
The number of legal and nonlegal ethical regulations in the biomedical field has increased tremendously, leaving present-day practitioners and researchers in a virtual crossfire of legislations and guidelines. Judging by the production and by the way these regulations are motivated and presented, they are held to be of great importance to ethical practice. This view is shared by many commentators. For instance, Commons and Baldwin write that, within the nursing profession, patient care can be performed unethically or ethically depending on the professional standards the nurses have set for themselves. They also hold that such standards are set when nurses become aware of the ethical codes available. As nurses are often not familiar with the codes, they do not all conform to them. Commons and Baldwin argue that nurses' ability to deal with ethical dilemmas is effectively secured with education on guidelines, creating a "barrier" between personal and professional values (p. 5).


Modality-specific and amodal aspects of object perception in infancy: the case of active touch.
Three experiments investigated 4.5-month-old infants' perception of the unity and boundaries of haptically presented objects. When infants actively explored the two handles of an unseen object assembly, perception of the unity of the assembly depended on the handles' motion. Infants perceived a single, connected object if the handles moved rigidly together, and they perceived two distinct objects if the handles underwent relative vertical or horizontal motion. When infants passively explored the same object assembly undergoing the same motions, object perception appeared to be indeterminate. The findings of the active motion experiments accord with the findings of studies of visual object perception and suggest that object perception depends on amodal processes, operating on representations of either seen or felt surface motions. The findings of the passive motion experiments nevertheless suggest a difference between visual and haptic perception: for infants as for adults, haptic perception is enhanced by the active production of surface motion.


The Salmonella Typhimurium effector protein SopE transiently localizes to the early SCV and contributes to intracellular replication.
Salmonella enterica serovar Typhimurium (S. Tm) is a facultative intracellular pathogen that induces entry into non-phagocytic cells by a Type III secretion system (TTSS) and cognate effector proteins. Upon host cell entry, S. Tm expresses a second TTSS and subverts intracellular trafficking to create a replicative niche - the Salmonella-containing vacuole (SCV). SopE, a guanidyl exchange factor (GEF) for Rac1 and Cdc42, is translocated by the TTSS-1 upon host cell contact and promotes entry through triggering of actin-dependent ruffles. After host cell entry, the bulk of SopE undergoes proteasomal degradation. Here we show that a subfraction is however detectable on the nascent SCV membrane up to ∼ 6 h post infection. Membrane localization of SopE and the closely related SopE2 differentially depend on the Rho-GTPase-binding GEF domain, and to some extent involves also the unstructured N-terminus. SopE localizes transiently to the early SCV, dependent on continuous synthesis and secretion by the TTSS-1 during the intracellular state. Mutant strains lacking SopE or SopE2 are attenuated in early intracellular replication, while complementation restores this defect. Hence, the present study reveals an unanticipated role for SopE and SopE2 in establishing the Salmonella replicative niche, and further emphasizes the importance of entry effectors in later stages of host-cell manipulation.


Acceptance, effects, and tolerability in the vaccination process against SARS-CoV-2 among cancer patients in Bosnia and Herzegovina: A single-center cross-sectional study.
The SARS-CoV-2 pandemic has been the main public health issue since the end of 2019. The vaccination campaign in Bosnia and Herzegovina started in April 2021, with several vaccines available. Our study aimed to evaluate the acceptance, effects, and tolerability of vaccines against SARS-COV-2 among cancer patients. We conducted a cross-sectional, observational study between 22 October and 30 November 2021, at the Clinic of Oncology, Clinical Center University of Sarajevo. Patients were enrolled during their regular visit to the Clinic of Oncology by agreeing to completean individual paper questionnaire. The study included 1063 patients with malignant diseases, of whom 681 (64.1%) were adequately vaccinated patients. In the study population, 76.9% of patients reported that they did not experience any side effects due to vaccination, while only 0.5% had side effects, causing a delay in their treatment. Among adequately vaccinated patients, there were 40 patients (3.8%) who were infected with SARS-CoV-2 after the second or booster dose of the vaccine. Five patients (0.5%) were hospitalized due to COVID-19 after being adequately vaccinated. The findings of our study suggest that cancer patients have a higher acceptance of vaccines against SARS-CoV-2 than the general population in Bosnia and Herzegovina. Vaccination side effects are tolerable and do not cause major delays in specific cancer treatment. The protective effects of COVID-19 vaccines in the cancer patients presented in our study are comparable to available results of similar studies, which included the general population.


Three-dimensional printed surgical templates for fresh cadaveric osteochondral allograft surgery with dimension verification by multivariate computed tomography analysis.
The fit of the allograft is a particular concern in fresh cadaveric osteochondral allograft (FOCA) surgery. Digital design and fabrication were utilized in conjunction with traditional surgery to enable efficient discovery and reproduction of appropriately dimensioned allograft. A patient with large osteochondral defects in the lateral femoral condyle was to undergo FOCA surgery. A digital virtual operation was performed, based on computed tomography (CT) images of the patient. Polyamide saw templates were manufactured using a selective laser sintering process, and gypsum powder was used to manufacture preoperative and intraoperative medical models with binder jetting process. The design dimensions were verified numerically by determining the intactness of the section surface and allograft volume based on four independent measurements of the initial design, and an automated design optimization strategy was postulated. For the surgery, a lateral longitudinal approach was employed. The virtual operation allowed an efficient design of the saw templates. Their shape and dimensions were verified with a numerical CT analysis method. The allograft dimensions (medial-lateral/superior-inferior/anterior-posterior) were approximately 40/28.5/24 mm, respectively, with the anterosuperior corner diagonally removed, yielding a section volume of approximately 16.5 cm3. These manually chosen dimensions were reminiscent of the corresponding computationally optimized values. Use of computer-aided design in virtual operation planning and three-dimensional printing in the fabrication of designed templates allowed for an efficient FOCA procedure and accurate allograft fitting. The numerical optimization method allowed for a semiautomated design process, which could in turn be realized also with surgical navigation or robotic surgery methods.


A long-term controlled follow-up study of objective treatment need on young adults treated with functional appliances.
The aims of this study were to 1) evaluate the objective success rate of Class II malocclusion treatment with functional appliances five years after completion of treatment and 2) to compare the remaining objective treatment need with an untreated control group. Records of all listed patients between 18-20 years (n=1054) treated in a general practice were reviewed for the purpose of finding treatments with removable functional appliances. Among all subjects (n=61) who previously had been treated, 58 accepted to participate in the study.The test group was matched with an orthodontically untreated group with no history of objective treatment need. Clinical examination was performed and study casts and photos were taken from both groups.The objective treatment need was evaluated through clinical examination and study cast analysis with weighted Peer Assessment Rating index (wPAR). Twenty patients, (34.5%) (mean wPAR 13.8), succeeded with the functional appliance treatment.The wPAR score (mean 15.0) of the entire test group was significantly higher than the one of the control group (mean 7.3).The group that was treated exclusively with functional appliances had a mean wPAR score of 17.4. Eighteen patients (31.0%) who received retreatment with fixed appliances had a slightly higher mean wPAR (8.6) than the control group. Treatments with functional appliances in a general practice showed a high failure rate and a remaining treatment need. It is the treating dentist's responsibility to motivate the patient to cooperate to the treatment, because as it previously has been shown the treatment with functional appliances is a well-functioning treatment alternative with the cooperation of the patient being sufficient. It is also of importance, already before starting treatment, to estimate the child's cooperation ability and to avoid treatment with removable appliances if the child or parents are reluctant about such a treatment.


Single inhaler extrafine triple therapy versus long-acting muscarinic antagonist therapy for chronic obstructive pulmonary disease (TRINITY): a double-blind, parallel group, randomised controlled trial.
Limited data are available for the efficacy of triple therapy with two long-acting bronchodilators and an inhaled corticosteroid in chronic obstructive pulmonary disease (COPD). We compared treatment with extrafine beclometasone dipropionate, formoterol fumarate, and glycopyrronium bromide (BDP/FF/GB; fixed triple) with tiotropium, and BDP/FF plus tiotropium (open triple). For this double-blind, parallel-group, randomised, controlled trial, eligible patients had COPD, post-bronchodilator forced expiratory volume in 1 s (FEV1) of less than 50%, at least one moderate-to-severe COPD exacerbation in the previous 12 months, and a COPD Assessment Test total score of at least 10. After a 2-week run-in period receiving one inhalation per day via single-dose dry-powder inhaler of open-label 18 μg tiotropium, patients were randomised (2:2:1) using a interactive response technology system to 52 weeks treatment with tiotropium, fixed triple, or open triple. Randomisation was stratified by country and severity of airflow limitation. The primary endpoint was moderate-to-severe COPD exacerbation rate. The key secondary endpoint was change from baseline in pre-dose FEV1 at week 52. The trial is registered with ClinicalTrials.gov, number NCT01911364. Between Jan 21, 2014, and March 18, 2016, 2691 patients received fixed triple (n=1078), tiotropium (n=1075), or open triple (n=538). Moderate-to-severe exacerbation rates were 0·46 (95% CI 0·41-0·51) for fixed triple, 0·57 (0·52-0·63) for tiotropium, and 0·45 (0·39-0·52) for open triple; fixed triple was superior to tiotropium (rate ratio 0·80 [95% CI 0·69-0·92]; p=0·0025). For week 52 pre-dose FEV1, fixed triple was superior to tiotropium (mean difference 0·061 L [0·037 to 0·086]; p<0·0001) and non-inferior to open triple (-0·003L [-0·033 to 0·027]; p=0·85). Adverse events were reported by 594 (55%) patients with fixed triple, 622 (58%) with tiotropium, and 309 (58%) with open triple. In our TRINITY study, treatment with extrafine fixed triple therapy had clinical benefits compared with tiotropium in patients with symptomatic COPD, FEV1 of less than 50%, and a history of exacerbations. Chiesi Farmaceutici SpA.


Daily consumption of pistachios over 12 weeks improves dietary profile without increasing body weight in healthy women: A randomized controlled intervention.
Dietary guidelines around the world recommend the regular intake of nuts because of their nutrient contribution to the diet and reported health benefits. However, consumers are often reluctant to include nuts in their diet due to the high caloric density of nuts. In a 12-week randomized, controlled intervention, we investigated how adding a pistachio portion to the daily diet affects body weight and composition, satiety, energy and nutrient intake. Sixty healthy pre-menopausal women who did not typically consume nuts were randomly assigned to a control or an experimental groups. Experimental participants (n = 30) consumed 44 g (250 kcal) pistachios mid-morning while controls (n = 30) maintained their current eating habits for 12 weeks. Ad libitum food intake and appetite sensations following the pistachio portion were assessed in all participants before and after the intervention in four 2-consecutive-day test sessions (under laboratory and then free-living conditions). Body weight and composition (DXA) were unchanged in both groups after 12 weeks. Pre- and post-intervention tests showed that ad libitum intake adjusted to the pistachio portion, mostly via reduced intakes of carbohydrates and starch, in parallel with decreased hunger and increased satiety following the morning snack. Intakes of MUFA, PUFA, linoleic acid, thiamin, pyridoxine, copper, manganese, and zinc were significantly higher among women consuming the pistachio snack, in spite of compensatory adjustments in intake. In conclusion, daily intake of 44 g pistachios improved nutrient intake without affecting body weight or composition in healthy women. The additional calories provided by the pistachios induced satiety and sufficient adjustment of intake to prevent body weight changes. CLINICAL TRIAL REGISTRY NUMBER: NCT03526120https://clinicaltrials.gov/ct2/show/NCT03526120.


The schistosome egg: development and secretions.
We have investigated the development of the schistosome egg and its secretions in order to understand how it migrates through gut tissues and also initiates pathology in the liver. We show by electron microscopy that the subshell envelope is absent in the newly deposited egg, but appears very early and differentiates as development progresses. In the mature egg, this nucleated envelope contains extensive endoplasmic reticulum, suggestive of a protein synthetic capacity. Furthermore, Reynolds' layer only appears between the envelope and the egg-shell in the mature egg and may represent its accumulated secretions. We have biosynthetically labelled and collected the secretions (ESP) released by mature but not immature eggs during culture. Their fractionation by SDS-PAGE reveals a simple pattern of 6 bands, differing markedly in composition from soluble egg antigen preparations. Electrophoresis in casein substrate gels demonstrates the presence of 2 distinct proteases in the egg secretions. By immunocytochemistry, ESP localized predominantly to the envelope of the mature egg, suggesting that this layer rather than the miracidium is the source of egg secretions.


Screening of phenylpyruvic acid producers and optimization of culture conditions in bench scale bioreactors.
Alpha keto acids are deaminated forms of amino acids that have received significant attention as feed and food additives in the agriculture and medical industries. To date, their production has been commonly performed at shake-flask scale with low product concentrations. In this study, production of phenylpyruvic acid (PPA), which is the alpha keto acid of phenylalanine was investigated. First, various microorganisms were screened to select the most efficient producer. Thereafter, growth parameters (temperature, pH, and aeration) were optimized in bench scale bioreactors to maximize both PPA and biomass concentration in bench scale bioreactors, using response surface methodology. Among the four different microorganisms evaluated, Proteus vulgaris was the most productive strain for PPA production. Optimum temperature, pH, and aeration conditions were determined as 34.5 °C, 5.12, and 0.5 vvm for PPA production, whereas 36.9 °C, pH 6.87, and 0.96 vvm for the biomass production. Under these optimum conditions, PPA concentration was enhanced to 1,054 mg/L, which was almost three times higher than shake-flask fermentation concentrations. Moreover, P. vulgaris biomass was produced at 3.25 g/L under optimum conditions. Overall, this study demonstrated that optimization of growth parameters improved PPA production in 1-L working volume bench-scale bioreactors compared to previous studies in the literature and was a first step to scale up the production to industrial production.


Whole-brain analysis of amyotrophic lateral sclerosis by using echo-planar spectroscopic imaging.
To detect regional metabolic differences in amyotrophic lateral sclerosis (ALS) with whole-brain echo-planar spectroscopic imaging. Sixteen patients with ALS (nine men, seven women; mean age, 56.6 years), five persons suspected of having ALS (four men, one woman; mean age, 62.6 years), and 10 healthy control subjects (five men, five women; mean age, 56.1 years) underwent echo-planar spectroscopic imaging after providing informed consent. The study was approved by the institutional review board and complied with HIPAA. Data were analyzed with the Metabolic Imaging and Data Analysis System software, and processed metabolite maps were coregistered and normalized to a standard brain template. Metabolite maps of creatine (Cr), choline (Cho), and N-acetylaspartate (NAA) were segmented into 81 regions with Automated Anatomical Labeling software to measure metabolic changes throughout the brains of patients with ALS. Statistical analysis involved an unpaired, uncorrected, two-sided Student t test. The NAA/Cho ratio across six regions was significantly lower by a mean of 23% (P ≤ .01) in patients with ALS than in control subjects. These regions included the caudate, lingual gyrus, supramarginal gyrus, and right and left superior and right inferior occipital lobes. The NAA/Cr ratio was significantly lower (P ≤ .01) in eight regions in the patient group, by a mean of 16%. These included the caudate, cuneus, frontal inferior operculum, Heschl gyrus, precentral gyrus, rolandic operculum, and superior and inferior occipital lobes. The Cho/Cr ratio did not significantly differ in any region between patient and control groups. Whole-brain echo-planar spectroscopic imaging permits detection of regional metabolic abnormalities in ALS, including not only the motor cortex but also several other regions implicated in ALS pathophysiologic findings.


Oscillatory electrodeposition of metal films at liquid/liquid interfaces induced by the large surface energy of growing deposits.
Electrodeposition of zinc (Zn) at an aqueous ZnSO4/n-butylacetate (BuAc) interface (liquid/liquid (LL) interface) showed a potential oscillation in the region of the current density exceeding the diffusion-limited one, accompanied by formation of two-dimensional Zn film with a concentric pattern at the LL interface. In-situ optical microscopic inspections revealed that the oscillatory growth of the Zn film synchronized with meniscus oscillation of the LL interface. The vigorous growth of the deposits occurs only when the shape of the meniscus becomes hollow on the negative potential side of the potential oscillation. On the other hand, on the positive side, the meniscus becomes almost flat and the deposits formed in the preceding stage are thickened. A mechanism is proposed to explain the oscillatory Zn electrodeposition coupled with the meniscus oscillation, on the basis of the fact that the interfacial tension at the growing metal/aqueous solution interface is extremely large.


Investigation of the causal relationships between human IgG N-glycosylation and 12 common diseases associated with changes in the IgG N-glycome.
Changes in the N-glycosylation of immunoglobulin G (IgG) are often observed in pathological states, such as autoimmune, inflammatory, neurodegenerative, cardiovascular diseases and some types of cancer. However, in most cases, it is not clear if the disease onset causes these changes, or if the changes in IgG N-glycosylation are among the risk factors for the diseases. The aim of this study was to investigate the casual relationships between IgG N-glycosylation traits and 12 diseases, in which the alterations of IgG N-glycome were previously reported, using two sample Mendelian randomization (MR) approach. We have performed two sample MR using publicly available summary statistics of genome-wide association studies of IgG N-glycosylation and disease risks. Our results indicate positive causal effect of systemic lupus erythematosus (SLE) on the abundance of N-glycans with bisecting N-acetylglucosamine in the total IgG N-glycome. Therefore, we suggest regarding this IgG glycosylation trait as a biomarker of SLE. We also emphasize the need for more powerful GWAS studies of IgG N-glycosylation to further elucidate the causal effect of IgG N-glycome on the diseases.


[Insomnia symptoms, daytime naps and physical leisure activities in the elderly: FIBRA Study Campinas].
The practice of physical activities contributes to reducing the risk of chronic diseases and improves sleep patterns in the elderly. This research aimed to investigate the association between insomnia symptoms and daytime nap and the participation in physical leisure activities in elderly community residents. Data from the Studies Network of the Fragility in Brazilian Elderly (Campinas site), were used. Information from 689 elderly was analyzed, regarding sociodemographic characterization, physical leisure activity, occurrence of daytime napping and its duration, symptoms of insomnia and use of sleep medication. A significant association was found between the practice of walking and the daytime nap of short duration. Studies indicate that a short nap can benefit the quality of sleep and health of the elderly. Therefore, promoting the practice of walking can be a nursing intervention that favors the sleep patterns of the elderly.


Localization of macromolecules in Escherichia coli. II. RNA and its site of synthesis.
The distribution of RNA in cells of E. coli 15 T(-)U(-) labeled with uridine-H(3) was studied by methods involving the analysis of radioautographic grain counts over random thin cross-sections and serial sections of the cells. The results were correlated with electron microscope morphological data. Fractionation and enzyme digestion studies showed that a large proportion of the label was found in RNA uracil and cytosine, the rest being incorporated as DNA cytosine. In fully labeled cells the distribution of label was found to be uniform throughout the cell. The situation remained unchanged when labeled cells were subsequently treated with chloramphenicol. When short pulses of label were employed a localization of a large proportion of the radioactivity became apparent. The nuclear region was identified as the site of concentration. Similar results were obtained when cells were exposed to much longer pulses of uridine-H(3) in the presence of chloramphenicol. If cells were subjected to a short pulse of cytidine-H(3), then allowed to grow for a while in unlabeled medium, the label, originally concentrated to some extent in the nuclear region, was found dispersed throughout the cell. The simplest hypothesis which accounts for these results is that a large fraction of the cell RNA is synthesized in a region in or near the nucleus and subsequently transferred to the cytoplasm.


The effect of health insurance and health facility-upgrades on hospital deliveries in rural Nigeria: a controlled interrupted time-series study.
Access to quality obstetric care is considered essential to reducing maternal and new-born mortality. We evaluated the effect of the introduction of a multifaceted voluntary health insurance programme on hospital deliveries in rural Nigeria. We used an interrupted time-series design, including a control group. The intervention consisted of providing voluntary health insurance covering primary and secondary healthcare, including antenatal and obstetric care, combined with improving the quality of healthcare facilities. We compared changes in hospital deliveries from 1 May 2005 to 30 April 2013 between the programme area and control area in a difference-in-differences analysis with multiple time periods, adjusting for observed confounders. Data were collected through household surveys. Eligible households ( n = 1500) were selected from a stratified probability sample of enumeration areas. All deliveries during the 4-year baseline period ( n = 460) and 4-year follow-up period ( n = 380) were included. Insurance coverage increased from 0% before the insurance was introduced to 70.2% in April 2013 in the programme area. In the control area insurance coverage remained 0% between May 2005 and April 2013. Although hospital deliveries followed a common stable trend over the 4 pre-programme years ( P = 0.89), the increase in hospital deliveries during the 4-year follow-up period in the programme area was 29.3 percentage points (95% CI: 16.1 to 42.6; P < 0.001) greater than the change in the control area (intention-to-treat impact), corresponding to a relative increase in hospital deliveries of 62%. Women who did not enroll in health insurance but who could make use of the upgraded care delivered significantly more often in a hospital during the follow-up period than women living in the control area ( P = 0.04). Voluntary health insurance combined with quality healthcare services is highly effective in increasing hospital deliveries in rural Nigeria, by improving access to healthcare for insured and uninsured women in the programme area.


The microRNA-124-iGluR2/3 pathway regulates glucagon release from alpha cells.
Glucagon, secreted from islet alpha cells, plays an important role in regulating glucose homeostasis; however, the molecular mechanism underlying this process is not fully understood. Previous studies have demonstrated that miRNAs are involved in the function of alpha cells. Glutamate promotes glucagon secretion by mediating the opening of Ca2+ channels. In this present, iGluR2 and iGluR3 levels were significantly increased in fasting-treated mouse islets. Additional studies showed that miR-124-3p simultaneously regulates the expression of iGluR2 and iGluR3 through the direct targeting of mRNA 3'UTR of these two genes. The miR-124-iGluRs pathway also contributed to the high level of glucagon secretion through long-term high glucose levels. Thus, a novel pathway comprising miRNA, glutamate and iGluRs has been demonstrated to regulate the biological process of glucagon release.


Clinical efficacy and survival analysis of apatinib combined with docetaxel in advanced esophageal cancer.
Standard chemotherapy has limited clinical efficacy in patients with esophageal cancer and there is a significant and unmet clinical need for effective treatment options for these patients. The aim of this study was to compare the clinical efficacy of the novel, targeted drug apatinib combined with docetaxel, and docetaxel combined with S-1 as second- or further-line treatment for patients with advanced esophageal cancer. We enrolled 33 patients with advanced esophageal cancer in chemotherapy group or apatinib combined with chemotherapy group in this retrospective study. Apatinib (500 mg) was taken orally once daily; docetaxel was administered at a dose of 75 mg/m2; and S-1 was optional at a dose of 40-60 mg, based on body surface area. The primary endpoint of this study was progression-free survival (PFS). Secondary endpoints included objective response rate (ORR), disease control rate (DCR), and the incidence and severity of adverse events (AEs). No complete response was observed in the two groups. However, two and five patients achieved partial response in the chemotherapy group and the apatinib combined with chemotherapy group, respectively. The ORR and DCR for the chemotherapy group was 11.1% and 33.3%, respectively. In the apatinib combination group, ORR and DCR was 88.9% and 93.3%, respectively. Anemia (11.1%) and neutropenia (5.6%) were the most frequent grade III/IV AEs observed in the chemotherapy group. In the apatinib combination group, the most frequent grade III/IV AEs were anemia (13.3%), hypertension (6.7%), and proteinuria (6.7%). Median PFS was significantly longer in the apatinib combination group than in the chemotherapy group (175 days vs 85 days, P=0.01). The combination of apatinib and docetaxel has a manageable toxicity profile and may prolong survival. Therefore, this combination may be used as as second- or further-line treatment for patients with advanced esophageal cancer.


Disruptions during a pandemic: Gaps identified and lessons learned.
Global disruptions caused by coronavirus disease 2019 (COVID-19) affects all walks of life, and dentistry and dental education are no exceptions. Dental education uniquely blends didactic courses and hands-on clinical training seamlessly to prepare oral healthcare providers of the future. Apart from economical and access to care implications, closure of all the dental institutions in the United States affects their educational mission greatly, equally disturbing pre-doctoral and graduate training. Efforts are ongoing to continue the educational mission in dental institutions by delivering scheduled course content remotely using multiple online tools. In spite of those efforts, since clinical experiences cannot be completely replaced by any available alternative method of instruction that is delivered remotely, students are missing out on valuable patient-based clinical experiences. In this perspective article, we briefly discuss the several implications of COVID-19, in the context of dental education. We then highlight some of the lessons we can learn from this pandemic which we hope will have several positive implications, including curricular changes, increased public health awareness and preparedness for future public health emergencies.


Adhesion of Histoplasma capsulatum to pneumocytes and biofilm formation on an abiotic surface.
The pathogenic fungus, Histoplasma capsulatum, causes the respiratory and systemic disease 'histoplasmosis'. This disease is primarily acquired via inhalation of aerosolized microconidia or hyphal fragments of H. capsulatum. Evolution of this respiratory disease depends on the ability of H. capsulatum yeasts to survive and replicate within alveolar macrophages. It is known that adhesion to host cells is the first step in colonization and biofilm formation. Some microorganisms become attached to biological and non-biological surfaces due to the formation of biofilms. Based on the importance of biofilms and their persistence on host tissues and cell surfaces, the present study was designed to investigate biofilm formation by H. capsulatum yeasts, as well as their ability to adhere to pneumocyte cells. H. capsulatum biofilm assays were performed in vitro using two different clinical strains of the fungus and biofilms were characterized using scanning electron microscopy. The biofilms were measured using a 2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-5-[(phenylamino)carbonyl]-2H-tetrazolium-hydroxide (XTT) reduction assay. The results showed that both the H. capsulatum strains tested were very efficient at adhering to host cells and forming biofilm. Therefore, this is a possible survival strategy adopted by this fungus.


Recovery of the gut microbiome following fecal microbiota transplantation.
Clostridium difficile infection is one of the most common health care-associated infections, and up to 40% of patients suffer from recurrence of disease following standard antibiotic therapy. Recently, fecal microbiota transplantation (FMT) has been successfully used to treat recurrent C. difficile infection. It is hypothesized that FMT aids in recovery of a microbiota capable of colonization resistance to C. difficile. However, it is not fully understood how this occurs. Here we investigated changes in the fecal microbiota structure following FMT in patients with recurrent C. difficile infection, and imputed a hypothetical functional profile based on the 16S rRNA profile using a predictive metagenomic tool. Increased relative abundance of Bacteroidetes and decreased abundance of Proteobacteria were observed following FMT. The fecal microbiota of recipients following transplantation was more diverse and more similar to the donor profile than the microbiota prior to transplantation. Additionally, we observed differences in the imputed metagenomic profile. In particular, amino acid transport systems were overrepresented in samples collected prior to transplantation. These results suggest that functional changes accompany microbial structural changes following this therapy. Further identification of the specific community members and functions that promote colonization resistance may aid in the development of improved treatment methods for C. difficile infection. Within the last decade, Clostridium difficile infection has surpassed other bacterial infections to become the leading cause of nosocomial infections. Antibiotic use, which disrupts the gut microbiota and its capability in providing colonization resistance against C. difficile, is a known risk factor in C. difficile infection. In particular, recurrent C. difficile remains difficult to treat with standard antibiotic therapy. Fecal microbiota transplantation (FMT) has provided a successful treatment method for some patients with recurrent C. difficile infection, but its mechanism and long-term effects remain unknown. Our results provide insight into the structural and potential metabolic changes that occur following FMT, which may aid in the development of new treatment methods for C. difficile infection.


The effect of chloramphenicol on deoxyribonucleic acid synthesis and the development of resistance to ultraviolet irradiation in E. coli infected with bacteriophage T2.
To elucidate the role of protein synthesis in DNA formation, E. coli R2 infected with phage T2 was studied as a model, employing chloramphenicol to inhibit protein synthesis. The following results were obtained. 1. Chloramphenicol inhibited protein synthesis but not synthesis of nucleic acids in uninfected bacteria. 2. Studies of the effect of chloramphenicol on phage maturation indicated a delay of 2 minutes between time of addition and cessation of phage growth. 3. The increase of DNA in phage-infected bacteria was completely suppressed by the addition of chloramphenicol within 2 minutes following infection. Addition at later times showed progressively less inhibitory action depending upon the time interval, and addition after the 10th or 12th minute showed no appreciable effect on DNA synthesis despite the cessation of intracellular phage formation and protein synthesis. 4. When chloramphenicol was added to infected cells the increase of resistance to UV stopped within 2 minutes, whether or not DNA synthesis continued. Thus evolution of resistance paralleled the rate of DNA synthesis achieved, but not the amount of DNA accumulated. 5. We conclude that in infected bacteria, protein synthesis is necessary to initiate DNA synthesis but is not essential for its continuation. The resistance to UV that characterizes infected cells near the midpoint of the latent period is not due to accumulation of DNA, but depends on some chloramphenicol-sensitive process (probably protein synthesis) completed at about the time the rate of DNA synthesis becomes maximal.


Combining mutual information with structural analysis to screen for functionally important residues in influenza hemagglutinin.
Influenza hemagglutinin mediates both cell-surface binding and cell entry by the virus. Mutations to hemagglutinin are thus critical in determining host species specificity and viral infectivity. Previous approaches have primarily considered point mutations and sequence conservation; here we develop a complementary approach using mutual information to examine concerted mutations. For hemagglutinin, several overlapping selective pressures can cause such concerted mutations, including the host immune response, ligand recognition and host specificity, and functional requirements for pH-induced activation and membrane fusion. Using sequence mutual information as a metric, we extracted clusters of concerted mutation sites and analyzed them in the context of crystallographic data. Comparison of influenza isolates from two subtypes--human H3N2 strains and human and avian H5N1 strains--yielded substantial differences in spatial localization of the clustered residues. We hypothesize that the clusters on the globular head of H3N2 hemagglutinin may relate to antibody recognition (as many protective antibodies are known to bind in that region), while the clusters in common to H3N2 and H5N1 hemagglutinin may indicate shared functional roles. We propose that these shared sites may be particularly fruitful for mutagenesis studies in understanding the infectivity of this common human pathogen. The combination of sequence mutual information and structural analysis thus helps generate novel functional hypotheses that would not be apparent via either method alone.


Exploring differential item functioning on eating disorder measures by food security status.
Food insecurity is associated with elevated eating disorder (ED) pathology, yet commonly used ED measures may not fully capture ED pathology in the context of food insecurity. The present study used differential item functioning (DIF) analyses to explore whether item endorsement on two commonly used ED questionnaires differed by food security status. Participants were 634 cisgender women recruited through Amazon's Mechanical Turk. DIF was explored for items on the Short Eating Disorder Examination Questionnaire (S-EDE-Q) and the Eating Disorder Diagnostic Scale for DSM-5 (EDDS-5). DIF analyses used a hybrid ordinal logistic regression/item response theory approach, with the presence of both statistical (p < .01) and clinical significance (pseudo ΔR2 ≥ .035) indicating DIF. There was no evidence of clinically significant DIF within the S-EDE-Q. Two items on the EDDS-5 exhibited statistically and clinically significant DIF, with moderate effect sizes. Specifically, compared to food-secure participants, food-insecure participants were more likely to report (1) eating large amounts of food when not physically hungry and (2) feeling disgusted, depressed, or guilty about overeating at lower levels of overall ED pathology but less likely to report these experiences at higher levels of overall ED pathology. Findings highlight a potential need to adapt ED measures to fully capture ED pathology in food-insecure populations. Level III, well-designed cohort study.


Oct4 is required ~E7.5 for proliferation in the primitive streak.
Oct4 is a widely recognized pluripotency factor as it maintains Embryonic Stem (ES) cells in a pluripotent state, and, in vivo, prevents the inner cell mass (ICM) in murine embryos from differentiating into trophectoderm. However, its function in somatic tissue after this developmental stage is not well characterized. Using a tamoxifen-inducible Cre recombinase and floxed alleles of Oct4, we investigated the effect of depleting Oct4 in mouse embryos between the pre-streak and headfold stages, ~E6.0-E8.0, when Oct4 is found in dynamic patterns throughout the embryonic compartment of the mouse egg cylinder. We found that depletion of Oct4 ~E7.5 resulted in a severe phenotype, comprised of craniorachischisis, random heart tube orientation, failed turning, defective somitogenesis and posterior truncation. Unlike in ES cells, depletion of the pluripotency factors Sox2 and Oct4 after E7.0 does not phenocopy, suggesting that ~E7.5 Oct4 is required within a network that is altered relative to the pluripotency network. Oct4 is not required in extraembryonic tissue for these processes, but is required to maintain cell viability in the embryo and normal proliferation within the primitive streak. Impaired expansion of the primitive streak occurs coincident with Oct4 depletion ∼E7.5 and precedes deficient convergent extension which contributes to several aspects of the phenotype.


Effect of basic fibroblast growth factor on perforated chinchilla tympanic membranes.
Basic fibroblast growth factor (bFGF) is a polypeptide mitogen which stimulates proliferation of epidermal and connective tissue cells. When applied to tympanic membrane perforations it has been reported to enhance healing and produce connective tissue hyperplasia. Previous work with animal models has shown that hyperplastic alterations of the tympanic membrane play an essential role in cholesteatoma development. This study was designed to further investigate the hyperplastic effects of bFGF and to determine if it might induce cholesteatoma formation during the healing process. Ten chinchillas received bilateral tympanic membrane perforations. In each animal, three doses of bFGF (400 nanograms per dose) were applied to the perforated tympanic membrane on one side; the opposite (control) ear received saline alone. The animals were terminated at either two or four weeks and studied histologically. Although the dosage and administration schedule used were consistent with previous studies utilizing other rodent species, there was little evidence that bFGF affected tympanic membrane healing in chinchillas. In both control and bFGF-treated ears, dense connective tissue occupied the lamina propria of the tympanic membrane, providing an effective barrier against ingrowth of skin toward the middle ear. No cholesteatomas developed in any animals included in the study. The results of this work indicate that the risk of cholesteatoma formation following administration of bFGF is minimal when it is applied short-term to acute perforations.


Skeletal muscle myosin heavy-chain synthesis rate in healthy humans.
Mixed muscle protein synthetic rate has been measured in humans. These measurements represent the average of synthetic rates of all muscle proteins with variable rates. We determined to what extent the synthesis rate of mixed muscle protein in humans reflects that of myosin heavy chain (MHC), the main contractile protein responsible for the conversion of ATP to mechanical energy as muscle contraction. Fractional synthetic rates of MHC and mixed muscle protein were measured from the increment of [13C]leucine in these proteins in vastus lateralis biopsy samples taken at 5 and 10 h during a primed continuous infusion of L-[1-13C]leucine in 10 young healthy subjects. Calculations were done by use of plasma [13C]ketoisocaproate (KIC) and muscle tissue fluid [13C]leucine as surrogate measures of leucyl-tRNA. Fractional synthetic rate of MHC with plasma KIC (0.0299 +/- 0.0043%/h) and tissue fluid leucine (0.0443 +/- 0.0056%/h) were only 72 +/- 3% of that of mixed muscle protein (0.0408 +/- 0.0032 and 0.0603 +/- 0.0059%/h, respectively, with KIC and tissue fluid leucine). Contribution of MHC (7 +/- 1 mg.kg-1.h-1) to synthetic rates of whole body mixed muscle protein (36 +/- 5 mg.kg-1.h-1) and whole body protein (127 +/- 4 mg.kg-1.h-1) is only 18 +/- 1 and 5 +/- 1%, respectively. This relatively low contribution of MHC to whole body and mixed muscle protein synthesis warrants direct measurement of synthesis rate of MHC in conditions involving abnormalities of muscle contractile function.


Self-reported STIs and sexual health checks in a cross-sectional study of gay and bisexual men in New Zealand.
To determine the incidence of self-reported sexually transmitted infections (STIs) and sexual health checks in community and internet samples of New Zealand gay, bisexual and other men who have sex with men (MSM) and factors associated with these. We analysed anonymous self-completed data from 3138 MSM who participated in the location-based Gay Auckland Periodic Sex Survey (GAPSS) and the internet-based Gay Online Sex Survey (GOSS) undertaken in February 2011. Overall 8.2% of the participants reported at least one STI in the previous 12 months, which did not differ significantly by demographic factors or HIV status. While having anal sex and more partners were associated with more STI, after adjustment for the number of partners, the type of partner (regular or casual) was not. Medium and low condom users reported STIs more than high condom users, regardless of partner type. Overall 40% had a sexual health check-up without an STI diagnosed in the past year, with similar numbers attending general practice and sexual health clinics. Having a check-up was lower among Pacific and Asian men, those identifying as bisexual and recruited online. While those with more partners, having anal intercourse and diagnosed with HIV were more likely to go for a check-up, those using condoms less often were not. STIs are commonly reported in this community sample of MSM but will underestimate the true incidence due to asymptomatic infection. Screening for STIs outside sexual health clinics should be normalised for MSM and made accessible, safe and relevant.


Trypanosoma brucei cathepsin-L increases arrhythmogenic sarcoplasmic reticulum-mediated calcium release in rat cardiomyocytes.
African trypanosomiasis, caused by Trypanosoma brucei species, leads to both neurological and cardiac dysfunction and can be fatal if untreated. While the neurological-related pathogenesis is well studied, the cardiac pathogenesis remains unknown. The current study exposed isolated ventricular cardiomyocytes and adult rat hearts to T. brucei to test whether trypanosomes can alter cardiac function independent of a systemic inflammatory/immune response. Using confocal imaging, T. brucei and T. brucei culture media (supernatant) caused an increased frequency of arrhythmogenic spontaneous diastolic sarcoplasmic reticulum (SR)-mediated Ca(2+) release (Ca(2+) waves) in isolated adult rat ventricular cardiomyocytes. Studies utilising inhibitors, recombinant protein and RNAi all demonstrated that this altered SR function was due to T. brucei cathepsin-L (TbCatL). Separate experiments revealed that TbCatL induced a 10-15% increase of SERCA activity but reduced SR Ca(2+) content, suggesting a concomitant increased SR-mediated Ca(2+) leak. This conclusion was supported by data demonstrating that TbCatL increased Ca(2+) wave frequency. These effects were abolished by autocamtide-2-related inhibitory peptide, highlighting a role for CaMKII in the TbCatL action on SR function. Isolated Langendorff perfused whole heart experiments confirmed that supernatant caused an increased number of arrhythmic events. These data demonstrate for the first time that African trypanosomes alter cardiac function independent of a systemic immune response, via a mechanism involving extracellular cathepsin-L-mediated changes in SR function.


Spontaneous Regression of Delayed Pulmonary and Mediastinal Metastases from Clear Cell Renal Cell Carcinoma.
Renal cell carcinoma (RCC) is often metastatic at diagnosis. Conventional therapies such as chemotherapy, radiotherapy, and hormonal therapy have generally proven ineffective in the treatment of RCC. The abscopal effect, specifically, the ability of localized radiation to trigger systemic antitumor effects, has been reported to lead to regression of non-irradiated distant tumor lesions. Herein, we report 3 patients with non-metastatic clear cell RCC (CCRCC) who underwent a nephrectomy and experienced metachronous pulmonary/mediastinal metastases confirmed as CCRCC. No patients underwent radiation post-nephrectomy or pulmonary metastasectomy. The mean duration was 7.24 weeks from the last negative chest CT prior to the nephrectomy and 96.2 weeks post-nephrectomy. All patients achieved durable complete response by RECIST criteria, with a mean follow-up duration of 115 months. Our case series represents the largest in the literature of patients who underwent a nephrectomy for CCRCC with no pre-existing pulmonary/mediastinal metastatic disease confirmed by chest CT, did not undergo radiotherapy, and developed significantly delayed CCRCC pulmonary/mediastinal metastases. We highlight the spontaneous regression of delayed metastatic disease and the role of immune responses in curtailing the growth of pulmonary metastasis in CCRCC.


Anti-MHC class I antibody activation of proliferation and survival signaling in murine cardiac allografts.
Anti-MHC class I alloantibodies have been implicated in the process of acute and chronic rejection because these Abs can bind to endothelial cells and transduce signals leading to the activation of cell survival and proliferation pathways. To characterize the role of the MHC class I-signaling pathway in the pathogenesis of Ab-mediated rejection, we developed a mouse vascularized heterotopic cardiac allograft model in which B6.RAG1 KO hosts (H-2K(b)/D(b)) received a fully MHC-incompatible BALB/c (H-2K(d)/D(d)) heart transplant and were passively transfused with anti-donor MHC class I Ab. We demonstrate that cardiac allografts of mice treated with anti-MHC class I Abs show characteristic features of Ab-mediated rejection including microvascular changes accompanied by C4d deposition. Phosphoproteomic analysis of signaling molecules involved in the MHC class I cell proliferation and survival pathways were elevated in anti-class I-treated mice compared with the isotype control-treated group. Pairwise correlations, hierarchical clustering, and multidimensional scaling algorithms were used to dissect the class I-signaling pathway in vivo. Treatment with anti-H-2K(d) Ab was highly correlated with the activation of Akt and p70S6Kinase (S6K). When measuring distance as a marker of interrelatedness, multidimensional scaling analysis revealed a close association between members of the mammalian target of rapamycin pathway including mammalian target of rapamycin, S6K, and S6 ribosomal protein. These results provide the first analysis of the interrelationships between these signaling molecules in vivo that reflects our knowledge of the signaling pathway derived from in vitro experiments.


Utility of MetaSite in improving metabolic stability of the neutral indomethacin amide derivative and selective cyclooxygenase-2 inhibitor 2-(1-(4-chlorobenzoyl)-5-methoxy-2-methyl-1H-indol-3-yl)-N-phenethyl-acetamide.
Prediction of the metabolic sites for new compounds, synthesized or virtual, is important in the rational design of compounds with increased resistance to metabolism. The aim of the present investigation was to use rational design together with MetaSite, an in silico tool for predicting metabolic soft spots, to synthesize compounds that retain their pharmacological effects but are metabolically more stable in the presence of cytochrome P450 (P450) enzymes. The model compound for these studies was the phenethyl amide (1) derivative of the nonsteroidal anti-inflammatory drug (NSAID) indomethacin. Unlike the parent NSAID, 1 is a potent and selective cyclooxygenase-2 (COX-2) inhibitor and nonulcerogenic anti-inflammatory agent in the rat. This pharmacological benefit is offset by the finding that 1 is very unstable in rat and human microsomes because of extensive P4503 A4/2D6-mediated metabolism on the phenethyl group, experimental observations that were accurately predicted by MetaSite. The information was used to design analogs with polar (glycinyl) and/or electron-deficient (fluorophenyl, fluoropyridinyl) amide substituents to reduce metabolism in 1. MetaSite correctly predicted the metabolic shift from oxidation on the amide substituent to O-demethylation for these compounds, whereas rat and human microsomal stability studies and pharmacokinetic assessments in the rat confirmed that the design tactics for improving pharmacokinetic attributes of 1 had worked in our favor. In addition, the fluorophenyl and pyridinyl amide derivatives retained the potent and selective COX-2 inhibition demonstrated with 1. Overall, the predictions from MetaSite gave useful information leading to the design of new compounds with improved metabolic properties.


What about the caregiver? A journey into Parkinson's disease following the burden tracks.
To investigate caregivers and patients characteristics related to different dimensions of burden in Parkinson's disease (PD). 55 pairs of PD patients and caregivers were recruited. The burden was evaluated with the Caregiver Burden Inventory (CBI). Multivariate analysis was applied to evaluate the impact of caregivers' and patients' characteristics on the varying aspects of burden. ADL score was the dominant predictor for the total score and all dimensions of CBI, except for the social burden, which is strongly predicted by the motor severity of PD. As one can easily imagine, the Total CBI decreases as the ADL score increases. An increased appreciation for characteristics of caregiver burden is a fundamental aspect of the patient's global evaluation. Clinicians may need to directly probe for these factors in the caregiver as they may not be elicited routinely.