[A rapid polycationic method for typing erythrocyte antibodies in liquid blood and stains].
The new rapid polycationic method for typing erythrocytic antibodies in liquid blood and its smears permits a rapid (within 4-5 min) detection of antibodies of virtually all erythrocytic isoserological systems ABO, MNSs, Rh, P, Lewis, Duffy, and Kidd in liquid blood and the group isohemagglutinins and incomplete Rh antibodies in blood smears. The test is performed on plane surface at ambient temperature and requires no special equipment. The sensitivity of the proposed method is much superior to that of the most sensitive and routine tests: enzymatic, antiglobulin, and salt. The only exclusion were anti-K antibodies, the majority of which were not detected by modern polycationic (tube and plane) methods.


Improved functional overview of protein complexes using inferred epistatic relationships.
Epistatic Miniarray Profiling(E-MAP) quantifies the net effect on growth rate of disrupting pairs of genes, often producing phenotypes that may be more (negative epistasis) or less (positive epistasis) severe than the phenotype predicted based on single gene disruptions. Epistatic interactions are important for understanding cell biology because they define relationships between individual genes, and between sets of genes involved in biochemical pathways and protein complexes. Each E-MAP screen quantifies the interactions between a logically selected subset of genes (e.g. genes whose products share a common function). Interactions that occur between genes involved in different cellular processes are not as frequently measured, yet these interactions are important for providing an overview of cellular organization. We introduce a method for combining overlapping E-MAP screens and inferring new interactions between them. We use this method to infer with high confidence 2,240 new strongly epistatic interactions and 34,469 weakly epistatic or neutral interactions. We show that accuracy of the predicted interactions approaches that of replicate experiments and that, like measured interactions, they are enriched for features such as shared biochemical pathways and knockout phenotypes. We constructed an expanded epistasis map for yeast cell protein complexes and show that our new interactions increase the evidence for previously proposed inter-complex connections, and predict many new links. We validated a number of these in the laboratory, including new interactions linking the SWR-C chromatin modifying complex and the nuclear transport apparatus. Overall, our data support a modular model of yeast cell protein network organization and show how prediction methods can considerably extend the information that can be extracted from overlapping E-MAP screens.


ICG-VA application in subtemporal transtentorial treatment of a Cognard V dural arteriovenous fistula.
Intracranial dural arteriovenous fistulas (dAVF) with cervical perimedullary drainage, Cognard V, are a surgically challenging rare entity. In this video we show the disconnection of a right tentorial Cognard V dAVF, done through a subtemporal transtentorial approach with the application of indocyanine green video angiography. A 47-year-old man presented with severe tetraparesis. Only partial embolization was possible. An osteoplastic frontotemporal craniotomy was performed to obtain a wide view along with CSF release to safely mobilize the temporal lobe. Neuronavigation was used to detect the fistula and indocyanine to detect the tentorial afferent arteries and to confirm final disconnection. The video can be found here: https://youtu.be/Yr8tAiiHNXU .


[Amylin under examination. Fibrillation - cytotoxic pancreatic polypeptide aggregation].
In patients or animals affected by type 2 diabetes mellitus (DM2, non-insulin dependent diabetes mellitus [NIDDM]), some pathological deposits, called amyloid, are observed among cells of islets of Langerhans. Among other constituents, the deposits consist of an insoluble, fibrillar form of polypeptide neurohormone called amylin, produced by pancreatic beta cells. It is thought that formation of fibrillar deposits of misfolded and aggregated polypeptide is highly toxic to beta cells and leads to cell dysfunction, cell loss, pancreas destruction and progress of the disease. Due to the extreme insolubility of this polypeptide and its instant fibrillation, amylin constitutes a methodological problem, and there is a need for a special methodology in experiments. Some mechanisms and factors that govern amylin fibrillization are rather poorly understood. This article presents amylin as a fibrillating molecule and some methods and methodological aspects and problems that emerge at successive steps during the fibrillation process, including hypothesized cytotoxicity mechanisms of this polypeptide.


Predictive Modeling and Concentration of the Risk of Suicide: Implications for Preventive Interventions in the US Department of Veterans Affairs.
The Veterans Health Administration (VHA) evaluated the use of predictive modeling to identify patients at risk for suicide and to supplement ongoing care with risk-stratified interventions. Suicide data came from the National Death Index. Predictors were measures from VHA clinical records incorporating patient-months from October 1, 2008, to September 30, 2011, for all suicide decedents and 1% of living patients, divided randomly into development and validation samples. We used data on all patients alive on September 30, 2010, to evaluate predictions of suicide risk over 1 year. Modeling demonstrated that suicide rates were 82 and 60 times greater than the rate in the overall sample in the highest 0.01% stratum for calculated risk for the development and validation samples, respectively; 39 and 30 times greater in the highest 0.10%; 14 and 12 times greater in the highest 1.00%; and 6.3 and 5.7 times greater in the highest 5.00%. Predictive modeling can identify high-risk patients who were not identified on clinical grounds. VHA is developing modeling to enhance clinical care and to guide the delivery of preventive interventions.


The importance of macroprolactinemia in the differential diagnosis of hyperprolactinemic patients.
Hypersecretion of prolactin (PRL) by lactotroph cells of the anterior pituitary may lead to hyperprolactinemia in physiological or pathological conditions. However, some of the patients may present with another cause of hyperprolactinemia, described by various authors as macroprolactinemia. The clinical, radiological and biochemical assessment of 124 patients were carefully evaluated for differential diagnosis in light of the literature. Macroprolactinemia was assessed by the polyethylene glycol (PEG) method in all of the patients, with high PRL level but without significant symptomatology, presenting to our clinic between 2004 and 2006. The sera from 124 patients with hyperprolactinemia were screened for macroprolactinemia using the PEG method and macroprolactinemia was detected in 10 patients (8%). The average age of the patients was 35 years (range 23-46). Nine of the ten patients were female (90%) and one was male (10%). All of the patients had MRI. An intrasellar mass and stalk lipoma were found in three of the ten patients (30%). In conclusion, macroprolactinemia should be taken into consideration as a probable cause of high serum prolactin levels to avoid repeated hormone assessments, neuroradiological examinations and unnecessary medical and surgical treatments.


[On the role of virus infection in the pathogenesis of vulvovaginitis in young girls].
The antigens of a number of widely prevalent viruses and changes typical of the cytopathic effect of these viruses were found in the epithelial cells of vaginal secretes in 44 out of 49 examined young girls with protracted forms of vulvovaginite. Mixed virus infection of the vaginal mucosa was diagnosed in 22 girls. More frequently than others, parainfluenza (57.1%) and adenovirus (59.2%) infections were detected in the patients. In girls of the control group, parainfluenza infection was diagnosed in 5.9% and adenovirus infection in 32.3%. Antigens of Coxsackie A and B (16.3%), influenza A2 and B (6.1%) and respiratory syncytial (6.1%) viruses were found in the vaginal mucosa in patients with vulvovaginitis only. A possible role of virus infection in the pathogenesis of vulvovaginitis in childhood is discussed.


[Access to oral health services in children under twelve years of age in Peru, 2014].
The aim of the study was to explore the patterns of dental health services access in children under twelve years of age in Peru. Data from 25,285 children under 12 years who participated in the Demographic and Family Health Survey of 2014 were reviewed. An exploratory spatial analysis was performed to project the proportions of children with access to dental health services, according to national regions, type of health service and urban or rural place of residence. The results show that of the total sample, 26.7% had access to dental health services in the last six months, 39.6% belonged to the age group 0-4 years, 40.6% lived in the Andean region and 58.3% lived in urban areas. The regions of Huancavelica, Apurimac, Ayacucho, Lima and Pasco had the highest percentages of access nationwide. In conclusion, there is low access to dental health services in the population under 12 years of age in Peru. The spatial distribution of access to dental health services allows regions to be identified and grouped according to similar access patterns, in order to better focus public health actions.


Homicidal methanol poisoning in a child.
A case of homicidal poisoning of a 21-month-old child using methanol is presented. The child was found dead in his crib during a court-ordered visit to his father's home. He reportedly was experiencing "flu-like" symptoms the day before. Routine toxicology testing of specimens taken at autopsy revealed the presence of methanol in a concentration of 0.21%, 0.23%, 0.31%, 0.28%, and 0.26% (w/v or w/w) in heart blood, venous blood, urine, vitreous humor, and gastric contents, respectively. Formic acid concentrations were 1.0 and 6.4 g/L in heart blood and urine. No other drugs or chemicals were detected in comprehensive screening. Accidental ingestion of methanol was ruled out; however, the homicide investigation was complicated by the fact that up to seven adults at three different locations had been involved in the child's care. Minimal information in the literature on the time frame for the development of symptoms of methanol intoxication in a child of this age made predicting a likely time of ingestion difficult. Discussion of the investigation and the timeline for the poisoning that was eventually established are included.


Prostate-specific antigen dynamics predict individual responses to intermittent androgen deprivation.
Intermittent androgen deprivation therapy (IADT) is an attractive treatment for biochemically recurrent prostate cancer (PCa), whereby cycling treatment on and off can reduce cumulative dose and limit toxicities. We simulate prostate-specific antigen (PSA) dynamics, with enrichment of PCa stem-like cell (PCaSC) during treatment as a plausible mechanism of resistance evolution. Simulated PCaSC proliferation patterns correlate with longitudinal serum PSA measurements in 70 PCa patients. Learning dynamics from each treatment cycle in a leave-one-out study, model simulations predict patient-specific evolution of resistance with an overall accuracy of 89% (sensitivity = 73%, specificity = 91%). Previous studies have shown a benefit of concurrent therapies with ADT in both low- and high-volume metastatic hormone-sensitive PCa. Model simulations based on response dynamics from the first IADT cycle identify patients who would benefit from concurrent docetaxel, demonstrating the feasibility and potential value of adaptive clinical trials guided by patient-specific mathematical models of intratumoral evolutionary dynamics.


Characterization of keratins from rat cervical epithelial cells in vivo and in vitro.
The keratins expressed in cultured rat cervical epithelial cells were analyzed by using two-dimensional gel electrophoresis and monoclonal antikeratin antibodies and were compared with those expressed in the rat cervix in vivo in the presence and absence of estrogen stimulation. The cervical epithelium in vivo responds to estrogen stimulation with alterations in keratin composition. In response to estrogen, the epithelium undergoes proliferation and stratification and begins expressing increased amounts of two basic Mr 57,000 and 58,000 keratins and two acidic Mr 51,000 and 52,000 keratins. Cultured rat cervical epithelial cells were found to express only small amounts of the basic Mr 57,000 and 58,000 keratins and the two acidic Mr 51,000 and 52,000 keratins characteristic of the estrogen-stimulated cervix. In addition to the keratins found in the cervical epithelium in vivo, rat cervical epithelial cells in vitro begin expressing two Mr 55,000 and 56,000 keratins of the basic keratin family (type II) and an acidic component with a molecular weight of 40,000. Although these components are not detected in the rat cervix in vivo, they migrate similarly on two-dimensional electrophoretic gels to proteins expressed by the rat endometrial epithelium in vivo. These findings indicate that alterations in keratin expression could serve as markers for studying the effects of steroid hormones on the differentiation of rat cervical epithelial cells in vitro and during the development of squamous metaplasia.


Cost-effectiveness of oxaliplatin in the adjuvant treatment of colon cancer in Canada.
The cost-effectiveness of oxaliplatin in combination with 5-fluorouracil/leucovorin (5FU/LV)-the FOLFOX regimen-was compared with that of 5FU/LV alone as adjuvant therapy for patients with stage III colon cancer, from the perspective of the Cancer Care Ontario New Drug Funding Program. In the mosaic (Multicenter International Study of Oxaliplatin/5-Fluorouracil/Leucovorin in the Adjuvant Treatment of Colon Cancer) trial, the FOLFOX regimen significantly improved disease-free survival. The mosaic trial formed the basis of the present analysis. Extrapolated patient-level data from the mosaic trial were used to model patient outcomes from treatment until death. Utilities were obtained from the literature. Resource utilization data were derived from the mosaic trial and supplemented with data from the literature. Unit costs were obtained from the Ontario Ministry of Health and Long-Term Care, the London Health Sciences Centre, and the literature. Lifetime incremental cost-effectiveness ratios for FOLFOX compared with 5fu/lv were CA$14,266 per disease-free year, CA$23,598 per life-year saved, and CA$24,104 per quality adjusted life-year (QALY) gained, discounting costs and outcomes at 5% per annum. These results were stable for a wide range of inputs; only utility values associated with relapse seemed to influence the cost-effectiveness ratios observed. With an incremental cost of CA$24,104 per QALY gained, FOLFOX is a cost-effective adjuvant treatment for stage iii colon cancer. Compared with 5fu/lv alone, this regimen offers better clinical outcomes and provides good value for money.


Evaluating the effects of vitamin D and vitamin E supplement on premenstrual syndrome: A randomized, double-blind, controlled trial.
Premenstrual syndrome (PMS) can cause problems in daily work and relationships. Eighty-six women were randomly assigned to two intervention groups and one control group. Patients were asked to fill out the PMS Daily Symptoms Record for 2 months, and then the participants were randomly assigned to one of the three study groups. Medical intervention was carried out for 2 months with the participants in each group receiving either a tablet containing 200 mg vitamin D, 100 mg vitamin E, or a placebo each day, respectively. After 2 months, the results of pre- and post-intervention were compared. P < 0.005 was considered significant. After the intervention, the mean score of the syndrome significantly decreased in all the three groups (12, 16, and 8 participants had decreased scores in vitamin D, vitamin E, and placebo, respectively). The differences between groups were not significant (P > 0.05). Supplemental therapy with vitamins D and E is an effective and affordable treatment for PMS.


Resistance to Src inhibition alters the BRAF-mutant tumor secretome to promote an invasive phenotype and therapeutic escape through a FAK>p130Cas>c-Jun signaling axis.
Few therapy options exist for patients with advanced papillary and anaplastic thyroid cancer. We and others have previously identified c-Src as a key mediator of thyroid cancer pro-tumorigenic processes and a promising therapeutic target for thyroid cancer. To increase the efficacy of targeting Src in the clinic, we sought to define mechanisms of resistance to the Src inhibitor, dasatinib, to identify key pathways to target in combination. Using a panel of thyroid cancer cell lines expressing clinically relevant mutations in BRAF or RAS, which were previously developed to be resistant to dasatinib, we identified a switch to a more invasive phenotype in the BRAF-mutant cells as a potential therapy escape mechanism. This phenotype switch is driven by FAK kinase activity, and signaling through the p130Cas>c-Jun signaling axis. We have further shown this more invasive phenotype is accompanied by alterations in the secretome through the increased expression of pro-inflammatory cytokines, including IL-1β, and the pro-invasive metalloprotease, MMP-9. Furthermore, IL-1β signals via a feedforward autocrine loop to promote invasion through a FAK>p130Cas>c-Jun>MMP-9 signaling axis. We further demonstrate that upfront combined inhibition of FAK and Src synergistically inhibits growth and invasion, and induces apoptosis in a panel of BRAF- and RAS-mutant thyroid cancer cell lines. Together our data demonstrate that acquired resistance to single-agent Src inhibition promotes a more invasive phenotype through an IL-1β>FAK>p130Cas>c-Jun >MMP signaling axis, and that combined inhibition of FAK and Src has the potential to block this inhibitor-induced phenotype switch.


Monoclonal Antibodies and Antivirals against SARS-CoV-2 Reduce the Risk of Long COVID: A Retrospective Propensity Score-Matched Case-Control Study.
Long COVID is a complex condition affecting quality of life, with limited therapeutic options. We investigated the occurrence of long COVID in subjects receiving early therapy with monoclonal antibodies (mAbs) or antivirals to reduce the risk of COVID-19 progression. In this retrospective study we enrolled 737 adult patients (aged 65.16 ± 13.46; 361F), who experienced COVID-19 between January 2021 and March 2022. Antiviral or mAbs were administered to symptomatic patients who did not require oxygen therapy or hospital admission for SARS-CoV-2 infection, and who were at high risk of progression to severe disease, as identified by age > 65 years or the presence of comorbidities. Long COVID, defined as newly or persistent long-term symptoms 4 weeks after the onset of the acute illness, was reported in 204 cases (28%). Age (OR 1.03; p < 0.001), gender (OR 1.88; p < 0.001) and at least three comorbidities (OR 3.49; p = 0.049) were directly associated with long COVID; conversely, vaccination (OR 0.59; p = 0.005) and mAbs/antivirals (OR 0.44; p = 0.002) were independently associated with a reduced risk of long COVID. At a propensity-score-matched analysis, the mAbs/antivirals group had a significantly lower occurrence of long COVID in comparison with untreated controls (11% vs. 34%; p = 0.001). In conclusion, mAbs and antivirals administered against the progression of COVID-19 were associated with a reduced risk of long COVID.


P-glycoprotein substrates and antagonists cluster into two distinct groups.
To gather further insight into the interaction between P-glycoprotein (Pgp) and its substrates, 167 compounds were analyzed in multidrug resistant human colon carcinoma cells. These compounds were selected from the National Cancer Institute Drug Screen repository using computer-generated correlations with known Pgp substrates and antagonists. The compounds were prospectively defined as Pgp substrates if cytotoxicity was increased > or =4-fold by the addition of cyclosporin A (CsA) and as Pgp antagonists if inhibition of efflux increased rhodamine accumulation by 4-fold. Among the 84 agents that met either criterion, 35 met only the criterion for substrates, 42 met only the criterion for antagonists, and only seven met both criteria. Thus, compounds interacting with Pgp form two distinct groups: one comprising cytotoxic compounds that are transported and have poor or no antagonistic activity and a second comprising compounds with antagonistic activity and no evidence of significant transport. Vinblastine accumulation and kinetic studies performed on a subset of 18 compounds similarly differentiated substrates and antagonists, but inhibition of 3H-azidopine labeling and induction of ATPase activity did not. These data support an emerging concept of Pgp in which multiple regions instead of specific sites are involved in drug transport.


Nurses' experience of adjusting to workplace violence: a theory of adaptation.
Workplace violence directed at nurses working in both the mental health and general areas of the hospital is a common occurrence and the impact of these events on all parties may be severe. A consequence of these confronting situations is the possibility that nurse victims will leave the profession. To help administrators facilitate nurses' psychological recovery, this qualitative study identified how nurses in several areas of a hospital setting adapted to workplace violence, research which has been previously unexamined. This study was the first of its kind to use a theory of cognitive adaptation to explore nurses' experiences of workplace violence. Participants were found to use the cognitive processes of finding meaning, gaining mastery and enhancing the self to adapt to workplace violence. Critical incident debriefing may facilitate the nurse victim's psychological recovery following an episode of workplace violence.


Talazoparib in Patients with Advanced Breast Cancer and a Germline BRCA Mutation.
The poly(adenosine diphosphate-ribose) inhibitor talazoparib has shown antitumor activity in patients with advanced breast cancer and germline mutations in BRCA1 and BRCA2 ( BRCA1/2). We conducted a randomized, open-label, phase 3 trial in which patients with advanced breast cancer and a germline BRCA1/2 mutation were assigned, in a 2:1 ratio, to receive talazoparib (1 mg once daily) or standard single-agent therapy of the physician's choice (capecitabine, eribulin, gemcitabine, or vinorelbine in continuous 21-day cycles). The primary end point was progression-free survival, which was assessed by blinded independent central review. Of the 431 patients who underwent randomization, 287 were assigned to receive talazoparib and 144 were assigned to receive standard therapy. Median progression-free survival was significantly longer in the talazoparib group than in the standard-therapy group (8.6 months vs. 5.6 months; hazard ratio for disease progression or death, 0.54; 95% confidence interval [CI], 0.41 to 0.71; P<0.001). The interim median hazard ratio for death was 0.76 (95% CI, 0.55 to 1.06; P=0.11 [57% of projected events]). The objective response rate was higher in the talazoparib group than in the standard-therapy group (62.6% vs. 27.2%; odds ratio, 5.0; 95% CI, 2.9 to 8.8; P<0.001). Hematologic grade 3-4 adverse events (primarily anemia) occurred in 55% of the patients who received talazoparib and in 38% of the patients who received standard therapy; nonhematologic grade 3 adverse events occurred in 32% and 38% of the patients, respectively. Patient-reported outcomes favored talazoparib; significant overall improvements and significant delays in the time to clinically meaningful deterioration according to both the global health status-quality-of-life and breast symptoms scales were observed. Among patients with advanced breast cancer and a germline BRCA1/2 mutation, single-agent talazoparib provided a significant benefit over standard chemotherapy with respect to progression-free survival. Patient-reported outcomes were superior with talazoparib. (Funded by Medivation [Pfizer]; EMBRACA ClinicalTrials.gov number, NCT01945775 .).


Rapid in vivo detection of rat spinal cord injury with double-diffusion-encoded magnetic resonance spectroscopy.
Diffusion-weighted imaging is a common experimental tool for evaluating spinal cord injury (SCI), yet it suffers from complications that decrease its clinical effectiveness. The most commonly used technique, diffusion tensor imaging (DTI), is often confounded by effects of edema accompanying acute SCI, limiting its sensitivity to the important functional status marker of axonal integrity. The purpose of this study is to introduce a novel diffusion-acquisition method with the goal of overcoming these limitations. A double diffusion encoding (DDE) pulse sequence was implemented with a diffusion-weighted filter orthogonal to the spinal cord for suppressing nonneural signals prior to diffusion weighting parallel to the cord. A point-resolved spectroscopy readout (DDE-PRESS) was used for improved sensitivity and compared with DTI in a rat model of SCI with varying injury severities. The DDE-PRESS parameter, restricted fraction, showed a strong relationship with injury severity (P < 0.001, R2 = 0.67). Although the whole-cord averaged DTI parameter values exhibited only minor injury relationships, a weighted region of interest (ROI) based DTI analysis improved sensitivity to injury (P < 0.001, R2 = 0.66). In a rat model of SCI, DDE-PRESS demonstrated high sensitivity to injury with substantial decreases in acquisition time and data processing. This method shows promise for application in rapid evaluation of SCI severity. Magn Reson Med 77:1639-1649, 2017. © 2016 International Society for Magnetic Resonance in Medicine.


In situ fluorescence spectroscopy correlates ionomer degradation to reactive oxygen species generation in an operating fuel cell.
The rate of generation of reactive oxygen species (ROS) within the polymer electrolyte membrane (PEM) of an operating proton exchange member fuel cell (PEMFC) was monitored using in situ fluorescence spectroscopy. A modified barrier layer was introduced between the PEM and the electrocatalyst layer to eliminate metal-dye interactions and fluorescence resonance energy transfer (FRET) effects during measurements. Standard fuel cell operating parameters (temperature, relative humidity, and electrode potential) were systematically varied to evaluate their influence on the rate of ROS generation during PEMFC operation. Independently, the macroscopic rate of PEM degradation was measured by monitoring the fluoride ion emission rate (FER) in the effluent stream at each operating condition. The ROS generation reaction rate constant (estimated from the in situ fluorescence experiments) correlated perfectly with the measured FER across all conditions, demonstrating unequivocally for the first time that a direct correlation exists between in situ ROS generation and PEM macroscopic degradation. The activation energy for ROS generation within the PEM was estimated to be 12.5 kJ mol(-1).


Sex differences in the experience of testing positive for genital chlamydia infection: a qualitative study with implications for public health and for a national screening programme.
To explore men's and women's accounts of chlamydia testing, with a view to understanding sex differences in attitudes and in behaviours of public health importance. Semistructured interviews with 24 heterosexual patients (12 men; 12 women) diagnosed with genital chlamydia infection, at a large sexual health clinic in central London were transcribed and analysed using qualitative thematic analysis. Participants' expressions revealed important sex differences. Women felt anxious about their future reproductive health, feared stigmatisation, and blamed themselves for contracting chlamydia; whereas men generally reported less concern, were unwilling to disclose their condition to sexual partners, and some men projected attributions of blame onto their partners. Delays in seeking care appeared to be related to perceptions of chlamydia as a relatively minor infection, particularly in men. Health promotion needs to reflect sex and age differences, emphasising the negative consequences of delayed clinic attendance and exposure to repeat infections. For health professionals to respond appropriately and effectively to increasing numbers of chlamydia infections, there is a need to understand men's avoidant attitudes and behaviours in relation to sexual and reproductive health.


Akt Regulates TNFα synthesis downstream of RIP1 kinase activation during necroptosis.
Necroptosis is a regulated form of necrotic cell death that has been implicated in the pathogenesis of various diseases including intestinal inflammation and systemic inflammatory response syndrome (SIRS). In this work, we investigated the signaling mechanisms controlled by the necroptosis mediator receptor interacting protein-1 (RIP1) kinase. We show that Akt kinase activity is critical for necroptosis in L929 cells and plays a key role in TNFα production. During necroptosis, Akt is activated in a RIP1 dependent fashion through its phosphorylation on Thr308. In L929 cells, this activation requires independent signaling inputs from both growth factors and RIP1. Akt controls necroptosis through downstream targeting of mammalian Target of Rapamycin complex 1 (mTORC1). Akt activity, mediated in part through mTORC1, links RIP1 to JNK activation and autocrine production of TNFα. In other cell types, such as mouse lung fibroblasts and macrophages, Akt exhibited control over necroptosis-associated TNFα production without contributing to cell death. Overall, our results provide new insights into the mechanism of necroptosis and the role of Akt kinase in both cell death and inflammatory regulation.


Acute Pulmonary Embolism in a Teenage Female - A Case Report.
Thrombophilia represents a tendency towards excessive blood clotting and the subsequent development of venous thromboembolism (VTE). VTE is a rare condition in children that comprises both deep venous thrombosis (DVT) and pulmonary embolism (PE). This paper reports the case of a 16-year-old girl, admitted to the Pediatrics Clinic No. 1, Tîrgu Mureș, Romania, for dyspnea, chest pain and loss of consciousness. Her personal history showed that she had had two orthopedic surgical interventions in infancy, two pregnancies, one spontaneous miscarriage and a recent caesarian section at 20 weeks of gestation for premature detachment of a normally positioned placenta associated with a deceased fetus. Laboratory tests showed increased levels of D-dimers. Angio-Computed Tomography (Angio-CT) showed multiple filling defects in both pulmonary arteries, establishing the diagnosis of PE. The laboratory tests were undertaken to assist in the diagnoses of a possible thrombophilia underlined a low level of antithrombin III. Antiphospholipid syndrome was ruled out and genetic tests revealed no specific mutation. Anticoagulant therapy was initiated with unfractionated heparin and afterwards subcutaneously low molecular heparin was prescribed for three months. Later it has been changed to oral therapy with acenocoumarol. The patient was discharged in good general status with the recommendation of life-long anticoagulation therapy. Thrombophilia is a significant risk factor for PE, and it must be ruled out in all cases of repeated miscarriage.


Impact of health beliefs, social support and self-efficacy on physical activity and dietary habits during the post-partum period after gestational diabetes mellitus: study protocol.
Gestational diabetes mellitus (GDM) is defined as a glucose intolerance of variable severity occurring or diagnosed for the first time during pregnancy. Numerous epidemiological studies show that this disorder affects between 1 and 18% of pregnancies, depending on the ethnicity of the populations studied, the diagnostic criteria, or the body mass index (BMI). Its incidence is constantly rising worldwide. Patients with GDM have a high risk of developing type 2 diabetes in the months after delivery. For this reason, GDM patients are encouraged to practice specific health behaviors (dietary habits, physical activity) during the postpartum period. It is important to identify the factors that may impact adherence to these behaviors. A targeted sample size of 200 eligible pregnant women with a diagnosis of GDM will be enrolled in this prospective, cohort study. They will be recruited from 30-36 weeks of gestation as part of their diabetes consultation in Geneva University Hospital (GUH) maternity unit. Psychosocial variables that could impact adherence to health behaviors in the postpartum period (behavioral intentions, risk perceptions, general knowledge about diabetes, health beliefs, social support, self-efficacy) will be evaluated using specific tools at the end of pregnancy, at 6 weeks postpartum and at 6 months postpartum. Multiple regression analyses will be performed on SPSS. For the first time in Europe, the objective of this research is to study in women with very recent GDM the link between dietary habits, physical activity levels, and psychosocial and cognitive factors possibly involved in the adoption of health behaviors in the postpartum period. These factors have been identified in the literature, but to date have never been combined in a single study. The study will allow a predictive theoretical model of health behavior to be established and used as a basis for reflection to optimize interventions carried out on women who have had GDM.


API ZYM and API An-Ident reactions of fastidious oral gram-negative species.
API ZYM and API An-Ident enzymatic substrate tests were done on six oral species which are difficult to characterize with conventional biochemical tests. "Bacteroides forsythus, the "fusiform" Bacteroides species (A. C. R. Tanner, M. A. Listgarten, M. N. Strzempko, and J. L. Ebersole, manuscript in preparation), is difficult to cultivate in broth media, yet it gave 15 positive tests in these series. The tests were able to separate this new species from species of Capnocytophaga and Fusobacterium. "B. forsythus" reactions were similar but not identical to those of reference Bacteroides species. Positive reactions for alpha-glucosidase, beta-glucosidase, alpha-fucosidase, and alpha-glucuronidase suggest that "B. forsythus" may be saccharolytic. It was the only species tested which was trypsin positive. Wolinella species, Campylobacter concisus, B. gracilis, and Eikenella corrodens are asaccharolytic, and characterization relies heavily on sensitivities to inhibitory agents. These species reacted weakly in the API ZYM and API An-Ident enzymatic substrate tests, and the reactions were not useful for separating these species. The enzyme reactions differentiated Wolinella recta and C. concisus from Selenomonas sputigena, another oral motile but saccharolytic organism.


170 W, single-frequency, single-mode, linearly-polarized, Yb-doped all-fiber amplifier.
A 170 W all-fiber linearly-polarized single-frequency sing-mode ytterbium amplifier at 1064 nm with an optical efficiency of 80% is demonstrated. 3.9 m long ytterbium-doped polarization maintaining fiber with a core diameter of 10 μm is used as the gain fiber, which guarantees a diffraction-limited output with a measured M(2) of 1.02. To suppress the stimulated Brillouin scattering, longitudinally varied strains are applied on the gain fiber according to the signal power evolution and the temperature distribution. 7 times increase of the stimulated Brillouin scattering threshold is achieved.


Obesity-Induced Changes in Bone Marrow Homeostasis.
Obesity is characterized by low-grade inflammation, which is accompanied by increased accumulation of immune cells in peripheral tissues including adipose tissue (AT), skeletal muscle, liver and pancreas, thereby impairing their primary metabolic functions in the regulation of glucose homeostasis. Obesity has also shown to have a detrimental effect on bone homeostasis by altering bone marrow and hematopoietic stem cell differentiation and thus impairing bone integrity and immune cell properties. The origin of immune cells arises in the bone marrow, which has been shown to be affected with the obesogenic condition via increased cellularity and shifting differentiation and function of hematopoietic and bone marrow mesenchymal stem cells in favor of myeloid progenitors and increased bone marrow adiposity. These obesity-induced changes in the bone marrow microenvironment lead to dramatic bone marrow remodeling and compromising immune cell functions, which in turn affect systemic inflammatory conditions and regulation of whole-body metabolism. However, there is limited information on the inflammatory secretory factors creating the bone marrow microenvironment and how these factors changed during metabolic complications. This review summarizes recent findings on inflammatory and cellular changes in the bone marrow in relation to obesity and further discuss whether dietary intervention or physical activity may have beneficial effects on the bone marrow microenvironment and whole-body metabolism.


A radiometric and petrographic approach to risk assessment at Alte Madonie Mounts region (Sicily, Italy).
The main goal of this work was to assess the radiological hazard at Alte Madonie Mounts region (north-central Sicily, Italy) in response to rumours of an increase in the incidence of cancer in this area. A correlation between the natural radionuclide contents and the petrographic features of the soil and rock samples was also evaluated. A total of 41 samples of selected soils and rocks were collected, powdered, dried and sealed in 'Marinelli' beakers for 20 d prior to measurement to ensure that a radioactive equilibrium between (226)Ra and (214)Bi had been reached. A gamma-ray spectrometer was used to quantify the radioactivity concentrations. To determine (238)U and (232)Th activities, the 609.3-keV line from (214)Bi in secular equilibrium with (226)Ra and the 911-keV line from (228)Ac, with which (232)Th can be assumed to be in equilibrium, were used, respectively. The gamma transition of 1461 keV was used to determine (40)K activity. The average values of the concentrations of (214)Bi, (228)Ac and (40)K were 30, 17 and 227 Bq kg(-1), respectively, whereas the greatest values were 134, 59 and 748 Bq kg(-1), respectively. A linear relationship was found between the activity values of (214)Bi, (228)Ac and (40)K. An exception was found for a group of samples in which the (214)Bi activities were much higher than expected. The chemical compositions and mineralogical features of the samples permitted the justification of these anomalies. The results of the primordial radionuclide contents are reassuring from a radiation protection point of view because the activities of the uranium and thorium series products and of the (40)K do not present a significant radiological hazard.


Clinical aspects and treatment of primary tracheal malignancies.
Selecting patients that are candidates for surgical treatment is important in the work-up of patients with tracheal cancer. Toward this goal, centralization of care concerning tracheal tumors is advised. Centralization may increase long-term survival and decrease operative morbidity and mortality even further. Primary tracheal tumors pose a diagnostic and therapeutic challenge for the physician when confronted with this mostly malignant tumor. Diagnosis is often delayed for months or years due to its aspecific and asthma-mimicking symptoms. Knowledge from retrospective series is limited and few clinicians have gained experience with this tumor. The available literature on the diagnosis and management of this group of tumors is reviewed to summarize the available knowledge about these uncommon tumors. New diagnostic, staging, and treatment guidelines are proposed. PubMed was searched for English publications from 1960. The available literature was reviewed and summarized. Surgical resection and primary reconstruction is the best curative treatment modality available at present. In centers of experience, more than half of all patients with tracheal cancer may be candidates for surgical resection, although in population-based studies this treatment is applied in only 10-25% of patients.


Characteristic Cytological Findings of Lobular Endocervical Glandular Hyperplasia Associated with Adenocarcinoma of the Uterine Cervix.
To investigate the cytological findings of lobular endocervical glandular hyperplasia (LEGH) associated with adenocarcinoma and to clarify its characteristics and the coexisting adenocarcinoma using histochemistry and immunohistochemistry. Eighteen surgical cases of LEGH of the uterine cervix were retrospectively reviewed and classified into 3 groups: pure (pure type), atypical (atypical type), and LEGH with adenocarcinoma (mixed type). The mixed type is defined as LEGH or atypical LEGH with in situ or invasive adenocarcinoma. Cytological findings of conventional endocervical smear specimens (Papanicolaou stain) were analyzed. Histochemistry (periodic acid-Schiff reaction) and immunohistochemistry (M-GGMC-1, Muc-6 glycoprotein, and Ki-67) were performed using tissue specimens. Cytologically, the pure type (7 cases) is characterized by glandular cell clusters that tended to form monolayered sheets with uniformly small nuclei and contain golden-yellowish mucin, whereas atypical (5 cases) and mixed (6 cases) types are characterized by glandular cell clusters similar to those of the pure type, but with complex glandular structures and mucin localization on the surface of glandular cell clusters. Ki-67 labeling index was significantly higher in atypical and mixed types than that in the pure type. Gastric-type mucinous carcinoma (MC-G) was observed in 2 out of 6 cases with mixed type. LEGH is found to be associated with adenocarcinoma types other than MC-G. Complex glandular structures or mucin localization on the surface of glandular cell clusters may be useful cytological findings to detect atypical and mixed types of LEGH.


[Tuberculin-dependent and nonspecific migration activity of leukocytes in astronauts].
There have been presented the results of studying the reactivity of leucocytes with respect to tuberculin and monspecific migration activity of leukocytes in the cosmonatus pre flight and after completion of orbital expeditions of various duration are presented. In some cosmonauts tuberculin rearranged migration of leukocytes. This may bear witness to activisation of the immunity system in response to the antigens which had been already familiar to test-subjects. Experimental data pointed to significant changes in leukocyte mobility in 12 out of 28 cosmonauts examined on days 1 and 7-14 of recovery after the first or repeated long-duration space missions indicating some functional disorders in the phagocytic link which may lead to a typical decline in human resistance to infections.


The pathogenesis of deferred cancer; a study of the after-effects of methylcholanthrene upon rabbit skin.
The ears of young adult rabbits were painted with methylcholanthrene (MC) long enough to call forth a few benign tumors (papillomas, frill horns), and the animals were followed throughout their later lives. Soon after the paintings were stopped the tumors began to dwindle and vanished, yet even while they were disappearing other growths of the same kinds arose, only to vanish later in their turn. For a long while more arose than disappeared, and in consequence the number of tumors increased throughout years. They accumulated at a constant rate despite concurrent changes in the supporting skin, which might have been supposed, on previous experience, to have prevented this from happening. Only in the old age of the animals did the number of tumors eventually fall off, and by this time the skin on which they had arisen, long since normal in the gross to all appearance, had become nearly so microscopically. Even then latent neoplastic potentialities still existed in the cutaneous tissue; where punch holes were healing new tumors arose. A great multitude of hidden neoplastic cells were present in the MCed skin, and from them many of the growths called forth by the stimulus of healing undoubtedly derived. Yet the facts make it difficult to suppose that the long accumulation, at a constant rate, of tumors visible in the gross was due wholly to the proliferation of cells rendered neoplastic during the period of exposure to MC, and lying hidden afterwards for periods determined by their differing, evenly graded capabilities. Nor can the accumulation be attributed to a sustained carcinogenesis resulting from the pathological state of the skin. As a whole the findings indicate that the linear increase in growths was due for the most part to a continual arrival at the neoplastic state and subsequent proliferation of cells, or the descendants of cells, that had been no more than started on the way toward becoming neoplastic by the carcinogen. There is clinical evidence for such a course of events. Now and again a carcinoma arose from the skin previously treated with MC, but they were few in all, as would follow from the presence of local conditions unfavorable to malignant change. Some appeared only after years,-in one instance more than 5 years after. The occurrence of deferred cancer in man can be understood in terms of the findings in rabbits.


Bereavement among South African adolescents following a sibling's death from AIDS.
While increasing attention has been paid in recent years to studying the impact of parental death from AIDS on children, we know little about how a sibling's death from AIDS affects children. In this qualitative descriptive study, 11 in-depth interviews were conducted by trained social workers with adolescents who had lost a sibling to AIDS, in KwaZulu-Natal Province, South Africa. The average time since the sibling's death was 18 months. These adolescents had been actively involved in caring for their sick sibling, yet they received inadequate emotional support from any source both before and after the sibling's death. HIV/AIDS stigma as well as the family's daily struggle to survive caused these adolescents to keep their feelings and their grief to themselves. Despite the trauma of losing a beloved sibling and the hardships of their environment, they demonstrated remarkable fortitude as well as concern for others. More research is needed on the issue of sibling bereavement associated with AIDS, and interventions that address the multiple needs of these vulnerable young people need to be developed.


Reduction of daily maintenance inhaled corticosteroids in patients with severe eosinophilic asthma treated with benralizumab (SHAMAL): a randomised, multicentre, open-label, phase 4 study.
Stepwise intensification of inhaled corticosteroids (ICS) is routine for severe eosinophilic asthma, despite some poor responses to high-dose ICS. Dose reductions are recommended in patients responding to biologics, but little supporting safety evidence exists. SHAMAL was a phase 4, randomised, open-label, active-controlled study done at 22 study sites in four countries. Eligible participants were adults (aged ≥18 years) with severe eosinophilic asthma and a five-item Asthma Control Questionnaire score below 1·5 and who received at least three consecutive doses of benralizumab before screening. We randomly assigned patients (3:1) to taper their high-dose ICS to a medium-dose, low-dose, and as-needed dose (reduction group) or continue (reference group) their ICS-formoterol therapy for 32 weeks, followed by a 16-week maintenance period. The primary endpoint was the proportion of patients reducing their ICS-formoterol dose by week 32. The primary outcome was assessed in the reduction group, and safety analyses included all randomly assigned patients receiving study treatment. This study is registered at ClinicalTrials.gov, NCT04159519. Between Nov 12, 2019, and Feb 16, 2023, we screened and enrolled in the run-in period 208 patients. We randomly assigned 168 (81%) to the reduction (n=125 [74%]) and reference arms (n=43 [26%]). Overall, 110 (92%) patients reduced their ICS-formoterol dose: 18 (15%) to medium-dose, 20 (17%) to low-dose, and 72 (61%) to as-needed only. In 113 (96%) patients, reductions were maintained to week 48; 114 (91%) of patients in the reduction group had zero exacerbations during tapering. Rates of adverse events were similar between groups. 91 (73%) patients had adverse events in the reduction group and 35 (83%) in the reference group. 17 patients had serious adverse events in the study: 12 (10%) in the reduction group and five (12%) in the reference group. No deaths occurred during the study. These findings show that patients controlled on benralizumab can have meaningful reductions in ICS therapy while maintaining asthma control. AstraZeneca.


Prospects for vaccination against pathogenic African trypanosomes.
African trypanosomes cause human and animal African trypanosomiases, which are chronic, debilitating and often fatal diseases of people and livestock in sub-Saharan Africa. The extracellular protozoan parasites are exemplars of antigenic variation. They direct host-protective B-cell and T-cell immune responses towards hypervariable components of their variable surface glycoprotein coat and evade immune elimination by generating new surface coat antigenic variants at a rate that supersedes immune destruction. This results in recurring waves of parasitemia, tissue invasion and escalating immunopathology in trypanosomiasis-susceptible hosts. Here, we discuss the possibility that host control of African trypanosomes might be improved by immunization with conserved VSG peptides and invariant surface glycoproteins. Infection-induced T-cell recall responses to these typically poorly expressed or nonimmunogenic parasite components induce tissue phagocytes to produce microbicidal materials that kill trypanosomes. Preliminary data that support this immune-enhancing vaccine strategy are discussed, as are host and parasite interactions that might downregulate the protective responses. These include infection-induced immunosuppression and increasing virulence of infecting parasites over time.


A data assimilation framework that uses the Kullback-Leibler divergence.
The process of integrating observations into a numerical model of an evolving dynamical system, known as data assimilation, has become an essential tool in computational science. These methods, however, are computationally expensive as they typically involve large matrix multiplication and inversion. Furthermore, it is challenging to incorporate a constraint into the procedure, such as requiring a positive state vector. Here we introduce an entirely new approach to data assimilation, one that satisfies an information measure and uses the unnormalized Kullback-Leibler divergence, rather than the standard choice of Euclidean distance. Two sequential data assimilation algorithms are presented within this framework and are demonstrated numerically. These new methods are solved iteratively and do not require an adjoint. We find them to be computationally more efficient than Optimal Interpolation (3D-Var solution) and the Kalman filter whilst maintaining similar accuracy. Furthermore, these Kullback-Leibler data assimilation (KL-DA) methods naturally embed constraints, unlike Kalman filter approaches. They are ideally suited to systems that require positive valued solutions as the KL-DA guarantees this without need of transformations, projections, or any additional steps. This Kullback-Leibler framework presents an interesting new direction of development in data assimilation theory. The new techniques introduced here could be developed further and may hold potential for applications in the many disciplines that utilize data assimilation, especially where there is a need to evolve variables of large-scale systems that must obey physical constraints.


SpHincterotomy for Acute Recurrent Pancreatitis Randomized Trial: Rationale, Methodology, and Potential Implications.
In patients with acute recurrent pancreatitis (ARP), pancreas divisum, and no other etiologic factors, endoscopic retrograde cholangiopancreatography (ERCP) with minor papilla endoscopic sphincterotomy (miES) is often performed to enlarge the minor papillary orifice, based on limited data. The aims of this study are to describe the rationale and methodology of a sham-controlled clinical trial designed to test the hypothesis that miES reduces the risk of acute pancreatitis. The SpHincterotomy for Acute Recurrent Pancreatitis (SHARP) trial is a multicenter, international, sham-controlled, randomized trial comparing endoscopic ultrasound + ERCP with miES versus endoscopic ultrasound + sham for the management of ARP. A total of 234 consented patients having 2 or more discrete episodes of acute pancreatitis, pancreas divisum confirmed by magnetic resonance cholangiopancreatography, and no other clear etiology for acute pancreatitis will be randomized. Both cohorts will be followed for a minimum of 6 months and a maximum of 48 months. The trial is powered to detect a 33% risk reduction of acute pancreatitis frequency. The SHARP trial will determine whether ERCP with miES benefits patients with idiopathic ARP and pancreas divisum. Trial planning has informed the importance of blinded outcome assessors and long-term follow-up.


Loss of IL-17-producing CD8 T cells during late chronic stage of pathogenic simian immunodeficiency virus infection.
Progressive disease caused by pathogenic SIV/HIV infections is marked by systemic hyperimmune activation, immune dysregulation, and profound depletion of CD4(+) T cells in lymphoid and gastrointestinal mucosal tissues. IL-17 is important for protective immunity against extracellular bacterial infections at mucosa and for maintenance of mucosal barrier. Although IL-17-secreting CD4 (Th17) and CD8 (Tc17) T cells have been reported, very little is known about the latter subset for any infectious disease. In this study, we characterized the anatomical distribution, phenotype, and functional quality of Tc17 and Th17 cells in healthy (SIV-) and SIV+ rhesus macaques. In healthy macaques, Tc17 and Th17 cells were present in all lymphoid and gastrointestinal tissues studied with predominance in small intestine. About 50% of these cells coexpressed TNF-α and IL-2. Notably, ∼50% of Tc17 cells also expressed the co-inhibitory molecule CTLA-4, and only a minority (<20%) expressed granzyme B suggesting that these cells possess more of a regulatory than cytotoxic phenotype. After SIV infection, unlike Th17 cells, Tc17 cells were not depleted during the acute phase of infection. However, the frequency of Tc17 cells in SIV-infected macaques with AIDS was lower compared with that in healthy macaques demonstrating the loss of these cells during end-stage disease. Antiretroviral therapy partially restored the frequency of Tc17 and Th17 cells in the colorectal mucosa. Depletion of Tc17 cells was not observed in colorectal mucosa of chronically infected SIV+ sooty mangabeys. In conclusion, our results suggest a role for Tc17 cells in regulating disease progression during pathogenic SIV infection.


What Is Ego Depletion? Toward a Mechanistic Revision of the Resource Model of Self-Control.
According to the resource model of self-control, overriding one's predominant response tendencies consumes and temporarily depletes a limited inner resource. Over 100 experiments have lent support to this model of ego depletion by observing that acts of self-control at Time 1 reduce performance on subsequent, seemingly unrelated self-control tasks at Time 2. The time is now ripe, therefore, not only to broaden the scope of the model but to start gaining a precise, mechanistic account of it. Accordingly, in the current article, the authors probe the particular cognitive, affective, and motivational mechanics of self-control and its depletion, asking, "What is ego depletion?" This study proposes a process model of depletion, suggesting that exerting self-control at Time 1 causes temporary shifts in both motivation and attention that undermine self-control at Time 2. The article highlights evidence in support of this model but also highlights where evidence is lacking, thus providing a blueprint for future research. Though the process model of depletion may sacrifice the elegance of the resource metaphor, it paints a more precise picture of ego depletion and suggests several nuanced predictions for future research.


Semi-autonomous surgical tasks using a miniature in vivo surgical robot.
Natural Orifice Translumenal Endoscopic Surgery (NOTES) is potentially the next step in minimally invasive surgery. This type of procedure could reduce patient trauma through eliminating external incisions, but poses many surgical challenges that are not sufficiently overcome with current flexible endoscopy tools. A robotic platform that attempts to emulate a laparoscopic interface for performing NOTES procedures is being developed to address these challenges. These robots are capable of entering the peritoneal cavity through the upper gastrointestinal tract, and once inserted are not constrained by incisions, allowing for visualization and manipulations throughout the cavity. In addition to using these miniature in vivo robots for NOTES procedures, these devices can also be used to perform semi-autonomous surgical tasks. Such tasks could be useful in situations where the patient is in a location far from a trained surgeon. A surgeon at a remote location could control the robot even if the communication link between surgeon and patient has low bandwidth or very high latency. This paper details work towards using the miniature robot to perform simple surgical tasks autonomously.


Lack of the inhibitory effect of intragastrically administered capsaicin on NNK-induced lung tumor formation in the A.J mouse.
Capsaicin is the principal component in Capsicum fruits consumed by humans worldwide as a food additive. The tobacco-specific nitrosamine 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) is thought to be an important causative factor in human lung cancer. Dietary factors are reported to modify lung tumor formation in laboratory animals and in humans. In this study, NNK-induced lung tumor formation in female A/J mice given intragastric doses of capsaicin (5 mg/kg body wt) was compared to mice not receiving capsaicin. At the end of the 21 week study, mice treated with capsaicin had an average of 17.1 +/- 1.8 lung tumors/mouse while untreated mice had 19.6 +/- 2. There were 100% lung tumor bearers in each group. Capsaicin alone did not affect spontaneous formation of lung tumors. Our results do not support a possible chemoprotective effect of dietary capsaicin toward NNK-induced lung tumors in human smokers.


An evaluation of the effectiveness of a community mentoring service for socially isolated older people: a controlled trial.
Social isolation affects a significant proportion of older people and is associated with poor health outcomes. The current evidence base regarding the effectiveness of interventions targeting social isolation is poor, and the potential utility of mentoring for this purpose has not previously been rigorously evaluated. The purpose of this study was to examine the effectiveness of a community-based mentoring service for improving mental health, social engagement and physical health for socially isolated older people. This prospective controlled trial compared a sample of mentoring service clients (intervention group) with a matched control group recruited through general practice. One hundred and ninety five participants from each group were matched on mental wellbeing and social activity scores. Assessments were conducted at baseline and at six month follow-up. The primary outcome was the Short Form Health Survey v2 (SF-12) mental health component score (MCS). Secondary outcomes included the SF-12 physical health component score (PCS), EuroQol EQ-5D, Geriatric Depression Score (GDS-10), social activity, social support and morbidities. We found no evidence that mentoring was beneficial across a wide range of participant outcomes measuring health status, social activity and depression. No statistically significant between-group differences were observed at follow-up in the primary outcome (p = 0.48) and in most secondary outcomes. Identifying suitable matched pairs of intervention and control group participants proved challenging. The results of this trial provide no substantial evidence supporting the use of community mentoring as an effective means of alleviating social isolation in older people. Further evidence is needed on the effectiveness of community-based interventions targeting social isolation. When using non-randomised designs, there are considerable challenges in the recruitment of suitable matches from a community sample. SCIE Research Register for Social Care 105923.


Delivering adolescent preventive services in an office setting.
Adolescence has long been recognized as a vulnerable period. Although several major medical professional organizations have published adolescent preventive care guidelines, regional and national surveys of youth, providers, and medical records all suggest many gaps exist in the delivery of recommended preventive services. In this article, we review the development of adolescent preventive health guidelines, describe current rates of preventive health screening and counseling, and explore barriers to delivery of preventive care for this age group. We present resources and practical suggestions to help the reader create an adolescent-friendly environment and to facilitate preventive health care delivery in your practice.


Stress induction and mitochondrial localization of Oxr1 proteins in yeast and humans.
Reactive oxygen species (ROS) are critical molecules produced as a consequence of aerobic respiration. It is essential for cells to control the production and activity of such molecules in order to protect the genome and regulate cellular processes such as stress response and apoptosis. Mitochondria are the major source of ROS within the cell, and as a result, numerous proteins have evolved to prevent or repair oxidative damage in this organelle. The recently discovered OXR1 gene family represents a set of conserved eukaryotic genes. Previous studies of the yeast OXR1 gene indicate that it functions to protect cells from oxidative damage. In this report, we show that human and yeast OXR1 genes are induced by heat and oxidative stress and that their proteins localize to the mitochondria and function to protect against oxidative damage. We also demonstrate that mitochondrial localization is required for Oxr1 protein to prevent oxidative damage.


Congenital dysfibrinogenemia in a Japanese family with fibrinogen Naples (BβAla68Thr) manifesting as superior sagittal sinus thrombosis.
: Congenital dysfibrinogenemia refers to the presence of a dysfunctional fibrinogen molecule, typically because of mutations in the fibrinogen gene. About 20% of fibrinogen gene mutations are responsible for thrombosis. Here, we described the case of a 17-year-old Japanese boy, who had a sudden stroke because of superior sagittal sinus thrombosis associated with dysfibrinogenemia. Genetic testing confirmed the presence of homozygous fibrinogen Naples (BβAla68Thr) mutation, which was previously reported as a causative mutation for thrombotic dysfibrinogenemia only in an Italian family. In this Japanese family, the patient's 12-year-old asymptomatic sister was also homozygous for this mutation. She, like her brother, was started on warfarin therapy. This report highlights the occurrence of fibrinogen Naples that has caused severe thrombotic complications in a young member of a Japanese family.


Impact on replicative fitness of the G48E substitution in the protease of HIV-1: an in vitro and in silico evaluation.
We observed an unusual glycine-to-glutamate substitution at protease (PR) residue position 48 (G48E) in an African patient infected with a subtype A1 HIV-1 strain failing a saquinavir-containing regimen. Phenotypic analysis of protease inhibitor (PI) susceptibility showed that the G48E site-directed mutant, when introduced into an NL4-3 HIV-1 PR backbone, was slightly resistant to SQV (2-fold when compared with the wild-type virus). In addition, the G48E and G48E/V82A site-directed mutants were associated with a decrease in fitness, whereas a reversion to the wild type at position 48 was observed in vitro. Growth competition experiments using a novel growth competition assay based on enhanced green fluorescent protein- or Discosoma spp. red fluorescent protein-expressing viruses showed that the replicative fitness of the G48E virus was reduced to 55% compared with the parental NL4-3 virus. Synthesizing all possible site-directed mutants found in the patient strain is too time-consuming; therefore, a molecular dynamics (MD) simulation approach was used to understand why this mutation survived despite its fitness cost. These simulations documented that the G48E mutant interacted with PI resistance mutations (M46I, I54V, Q58E, and L63P) and with natural polymorphisms specific to subtype A1 (E35D, M36I, and R57K) that were present in the patient's virus. We hypothesize that the polymorphisms contained in the PR flap regions of the patient's virus may compensate for the presence of G48E, possibly by restoring the flexibility of the PR flaps. In summary, our results demonstrate that the G48E substitution, when introduced in the context of an HIV-1 subtype B strain, is highly unstable and gives rise to viruses with a poor replicative fitness in vitro. We also showed that when confronted with too many mutations to evaluate in vitro, MD simulations are helpful to draft hypotheses on how polymorphisms can interact with resistance mutations to stabilize their potential fitness cost.


Targeting Angiogenesis in Pancreatic Neuroendocrine Tumors: Resistance Mechanisms.
Despite being infrequent tumors, the incidence and prevalence of pancreatic neuroendocrine tumors (P-NETs) has been rising over the past few decades. In recent years, rigorous phase III clinical trials have been conducted, allowing the approval of several drugs that have become the standard of care in these patients. Although various treatments are used in clinical practice, including somatostatin analogues (SSAs), biological therapies like sunitinib or everolimus, peptide receptor radionuclide therapy (PRRT) or even chemotherapy, a consensus regarding the optimal sequence of treatment has not yet been reached. Notwithstanding, sunitinib is largely used in these patients after the promising results shown in SUN111 phase III clinical trial. However, both prompt progression as well as tumor recurrence after initial response have been reported, suggesting the existence of primary and acquired resistances to this antiangiogenic drug. In this review, we aim to summarize the most relevant mechanisms of angiogenesis resistance that are key contributors of tumor progression and dissemination. Furthermore, several targeted molecules acting selectively against these pathways have shown promising results in preclinical models, and preliminary results from ongoing clinical trials are awaited.


Efficacy of the early treatment with tocilizumab-hydroxychloroquine and tocilizumab-remdesivir in severe COVID-19 Patients.
The effectiveness of the best combination between different antiviral and anti-inflammatory drugs stills an interest in the treatment of COVID19 infection. A prospective randomized cohort study comprised 108 adult patients with confirmed PCR COVID 19 infection with systemic hyper inflammation state, divided into two groups according to the treatment regimen, 56 in the tocilizumab- hydroxychloroquine (TCZ-HCQ) treatment, and 52 in the tocilizumab-remdesivir (TCZ-RMV) treatment. The first group received a combination of I.V. TCZ (400-800 mg every 24 h for only two doses) and HCQ (400 mg twice in the first day then 200 mg twice for 5 days) while the second group of patients received I.V. RMV of 200 mg on day 1 followed by 100 mg once daily infused over 60 min for 5 days with the same TCZ regimen used in the first group. All clinical parameters and laboratory investigations were assessed before and after treatment. The CRP was significantly decreased while PaO2/FiO2 (P/F) ratio post-treatment was significantly improved in both treatment groups. TCZ-HCQ group showed a significant decrease in the ferritin, LDH, and D. Dimer levels. The median days of hospitalization with interquartile range (IQR) were 10 (6-16) and 8 (5-12) for TCZ-HCQ and TCZ-RMV groups, respectively. The numbers of mechanically ventilated patients were 25 and 43 for TCZ-HCQ and TCZ-RMV groups, respectively. Therapeutic failure was about 26.8% in the TCZ-HCQ group and 30.8% in the TCZ-RMV group but there was no significant difference between both groups. Some complications were recognized only in TCZ-RMV following treatment including secondary bacterial infections (42.3%), myocarditis (15.4%), and finally pulmonary embolism (7.7%). Efficacy of both TCZ-RMV and TCZ-HCQ combinations are observed in the treatment of severe COVID-19 patients; however the increased need for ICU or mechanical ventilation in the TCZ-RMV arm contributed to the appearance of cardiac and thrombotic events. The study was registered at the Clinical Trials registry (ClinicalTrials.gov; NCT04779047).


[CUTANEOUS LEISHMANIASIS IN ISRAEL 2016 - AN UPDATE].
Leishmaniasis is endemic in Israel and is increasing in its frequency. The pathogens that cause cutaneous leishmaniasis (CL) in Israel are Leishmania major (L. major) and Leishmania tropica (L. tropica). In recent years, there have been significant changes in the distribution of these species. L. major is endemic mainly in the Negev, Arava and the Beit She'an Valley. L. tropica is endemic in Tiberias, northern Sea of Galilee, Samaria and in the vicinity of Jerusalem such as at Kfar Adumim, Ma'ale Adumim and Anathoth (north-east of Jerusalem). In addition, there is imported Leishmania Braziliensis (L. braziliensis) from Israeli travelers, originating mainly from the Amazon region of Bolivia. The clinical course of CL and the best treatment varies and is determined by the leishmania species. Among the endemic pathogens in Israel, the clinical course of L. tropica can be more severe, last longer and may be more resistant to treatment in comparison to L. major. However, imported leishmaniasis carries the risk of muco-cutaneous involvement and it is therefore important to identify the exact species to determine the appropriate treatment. The main treatment options for endemic cutaneous leishmaniasis are topical treatment with paromomycin ointment, liquid nitrogen, or intra-lesional injectable sodium stibogluconate (pentostam). Occasionally, there are cases of CL that require systemic therapy, such as in cases that lack response to local treatment, cases of multiple lesions or if affected areas of the body are such that local treatment cannot be applied. The most common systemic treatment is intravenous sodium stibogluconate (pentostam). This treatment may cause serious side effects. Therefore, in recent years we use intravenous liposomal amphotericin B (AmBisome) as the first line treatment, with a high success rate. Standard treatment for imported L. braziliensis is intravenous therapy in order to prevent its spread to the naso-pharynx, which can cause permanent destruction of the cartilage in these areas. The drug of choice is intravenous liposomal amphotericin B, and in cases of treatment failure, intravenous sodium stibogluconate (pentostam) can be used. This year the use of oral miltefosine tablets was also approved in Israel for cases of cutaneous leishmaniasis.


Partial migration in fishes: causes and consequences.
Partial migration, where only some individuals from a population migrate, has been widely reported in a diverse range of animals. In this paper, what is known about the causes and consequences of partial migration in fishes is reviewed. Firstly, the ultimate and proximate drivers of partial migration are reflected upon: what ecological factors can shape the evolution of migratory dimorphism? How is partial migration maintained over evolutionary timescales? What proximate mechanisms determine whether an individual is migratory or remains resident? Following this, the consequences of partial migration are considered, in an ecological and evolutionary context, and also in an applied sense. Here it is argued that understanding the concept of partial migration is crucial for fisheries and ecosystem managers, and can provide information for conservation strategies. The review concludes with a reflection on the future opportunities in this field, and the avenues of research that are likely to be fruitful to shed light on the enduring puzzle of partial migration in fishes.


Comparison of two contemporary rotary systems in a pre-clinical student course setting.
To assess two contemporary rotary instrumenting systems subjectively and objectively in a pre-clinical student course setting. Undergraduate dental students (n = 44) prepared mesiolingual canals of 3D-printed mandibular molar replicas (RepliDens, Zurich, Switzerland). The HyFlex and BioRace rotary systems, both previously unknown to the students, were used according to the manufacturers' guidelines after a short theoretical introduction. For comparison, a first-generation rotary system (ProFile .04), which the students knew from their previous education, was then used in a third RepliDens. Questionnaires were issued to note subjective experiences immediately after instrumentation. Objectively, time to instrument to size 40, .04 taper and shaping outcomes were analysed. Categorical data were compared using chi-square and Fisher's exact tests, numerical data according to goodness of fit to the normal distribution, P < 0.05. Subjectively, the students liked the file size and sequence designation in the BioRace system significantly (P < 0.05) better than in the HyFlex counterpart, whilst they found better controllability with the HyFlex (P < 0.05), and reported no difference in cutting efficiency. They preferred both systems to the ProFile. Objectively, canal transportation was significantly less with the HyFlex (and the ProFile) systems compared to BioRace (P < 0.05). Both systems under investigation were statistically similar in terms of file fractures (nil), length control, and instrumentation time, which was considerably faster than with the ProFile control system. HyFlex and BioRace had perceived and quantifiable strengths and weaknesses. Both systems were equally liked by the students and preferred over the ProFile first-generation rotary system.


A Practical Guide for Treatment of Pain in Patients with Systemic Mast Cell Activation Disease.
Systemic mast cell activation disease (MCAD, a subclass of mastocytosis), which has a prevalence of around 17% (at least in the German population), is characterized by accumulation of genetically altered dysfunctional mast cells with abnormal release of these cells' mediators. Since mast cells affect functions in potentially every organ system, often without causing abnormalities in routine laboratory or radiologic testing, this disease has to be considered routinely in the differential diagnosis of patients with chronic multisystem polymorbidity of a generally inflammatory and allergic theme. Pain in its different manifestations is a common symptom in MCAD found in more than three-quarters of the MCAD patients. Because of the specific mast cell-related causes of pain in MCAD it should be treated specifically, if possible, deduced from their putative mast cell mediator-related causes. As yet, there is no official guideline for treatment of MCAD at all. The present review focuses on mast cell mediator-induced acute and chronic pain and the current state of analgesic drug therapy options in MCAD. Due to the high prevalence of MCAD, many physicians are often faced with the issue of pain management in MCAD patients. Hence, our practical guide should contribute to the improvement of patient care.Key words: Pain therapy, mast cell activation disease, mast cell activation syndrome, systemic mastocytosis, mast cell.


Short-Term Exposure to Ambient Air Pollution and Influenza: A Multicity Study in China.
Air pollution is a major risk factor for planetary health and has long been suspected of predisposing humans to respiratory diseases induced by pathogens like influenza viruses. However, epidemiological evidence remains elusive due to lack of longitudinal data from large cohorts. Our aim is to quantify the short-term association of influenza incidence with exposure to ambient air pollutants in Chinese cities. Based on air pollutant data and influenza surveillance data from 82 cities in China over a period of 5 years, we applied a two-stage time series analysis to assess the association of daily incidence of reported influenza cases with six common air pollutants [particulate matter with aerodynamic diameter ≤2.5μm (PM2.5), particulate matter with aerodynamic diameter ≤10μm (PM10), NO2, SO2, CO, and O3], while adjusting for potential confounders including temperature, relative humidity, seasonality, and holiday effects. We built a distributed lag Poisson model for one or multiple pollutants in each individual city in the first stage and conducted a meta-analysis to pool city-specific estimates in the second stage. A total of 3,735,934 influenza cases were reported in 82 cities from 2015 to 2019, accounting for 72.71% of the overall case number reported in the mainland of China. The time series models for each pollutant alone showed that the daily incidence of reported influenza cases was positively associated with almost all air pollutants except for ozone. The most prominent short-term associations were found for SO2 and NO2 with cumulative risk ratios of 1.094 [95% confidence interval (CI): 1.054, 1.136] and 1.093 (95% CI: 1.067, 1.119), respectively, for each 10 μg/m3 increase in the concentration at each of the lags of 1-7 d. Only NO2 showed a significant association with the daily incidence of influenza cases in the multipollutant model that adjusts all six air pollutants together. The impact of air pollutants on influenza was generally found to be greater in children, in subtropical cities, and during cold months. Increased exposure to ambient air pollutants, particularly NO2, is associated with a higher risk of influenza-associated illness. Policies on reducing air pollution levels may help alleviate the disease burden due to influenza infection. https://doi.org/10.1289/EHP12146.


Efficacy of voriconazole and amphotericin B in corneal preservative media.
To evaluate the efficacy of voriconazole and amphotericin B in McCarey-Kaufman (MK) media. MK media vials were supplemented with either voriconazole at 1, 2, 20, 50, 100 μg/mL or amphotericin B at 0.5, 1, 2, 10, 20 μg/mL. The standard inoculum of the American Type Culture Collection (ATCC) strain of Candida albicans, Aspergillus flavus, and Fusarium keratinoplasticum was added to the set of vials. The efficacy outcomes were calculated as 'viable fungal colony counts' determined from the samples taken on Days 0 and 4. MK media containing fungal inoculum but without antifungal supplements were used as control. In the voriconazole arm, on Day 4, a reduction in the colony count was observed for Candida albicans (1 μg/mL, 36%; 100 μg/mL, 100%), Aspergillus flavus (1 μg/mL, 53.8%; 100 μg/mL, 80.4%), and Fusarium keratinoplasticum (1 μg/mL, 39.0%; 100 μg/mL, 72.2%). Similarly, in the amphotericin B arm, on Day 4, a reduction in the colony count was observed for Candida albicans (0.5 μg/mL; 99.9%; 20 μg/mL, 100%), Aspergillus flavus (0.5 μg/mL, 65.2%; 20 μg/mL, 84.8%), and Fusarium keratinoplasticum (0.5 μg/mL, 90.1%; 20 μg/mL, 100%). Compared to voriconazole, the addition of amphotericin B significantly reduces fungal contamination in MK media.


Prophylactic use of a probiotic in the prevention of colic, regurgitation, and functional constipation: a randomized clinical trial.
Infantile colic, gastroesophageal reflux, and constipation are the most common functional gastrointestinal disorders that lead to referral to a pediatrician during the first 6 months of life and are often responsible for hospitalization, feeding changes, use of drugs, parental anxiety, and loss of parental working days with relevant social consequences. To investigate whether oral supplementation with Lactobacillus reuteri DSM 17938 during the first 3 months of life can reduce the onset of colic, gastroesophageal reflux, and constipation in term newborns and thereby reduce the socioeconomic impact of these conditions. A prospective, multicenter, double-masked, placebo-controlled randomized clinical trial was performed on term newborns (age <1 week) born at 9 different neonatal units in Italy between September 1, 2010, and October 30, 2012. Parents were asked to record in a structured diary the number of episodes of regurgitation, duration of inconsolable crying (minutes per day), number of evacuations per day, number of visits to pediatricians, feeding changes, hospitalizations, visits to a pediatric emergency department for a perceived health emergency, pharmacologic interventions, and loss of parental working days. In total, 589 infants were randomly allocated to receive L reuteri DSM 17938 or placebo daily for 90 days. Prophylactic use of probiotic. Reduction of daily crying time, regurgitation, and constipation during the first 3 months of life. Cost-benefit analysis of the probiotic supplementation. At 3 months of age, the mean duration of crying time (38 vs 71 minutes; P < .01), the mean number of regurgitations per day (2.9 vs 4.6; P < .01), and the mean number of evacuations per day (4.2 vs 3.6; P < .01) for the L reuteri DSM 17938 and placebo groups, respectively, were significantly different. The use of L reuteri DSM 17938 resulted in an estimated mean savings per patient of €88 (US $118.71) for the family and an additional €104 (US $140.30) for the community. Prophylactic use of L reuteri DSM 17938 during the first 3 months of life reduced the onset of functional gastrointestinal disorders and reduced private and public costs for the management of this condition. clinicaltrials.gov Identifier: NCT01235884.


Synergistic effects of alpha interferon and 1,25 dihydroxyvitamin D3: preliminary evidence suggesting that interferon induces expression of the vitamin receptor.
To active metabolite of vitamin D3-1,25(OH)2D3-is a well-known differentiation inducer. The addition of this metabolite to sensitive cell cultures inhibits proliferation and induces monocytic-macrophagic differentiation. Alpha interferon may also inhibit proliferation and increase the expression of some surface antigens in some neoplastic cells. In the present report, we describe the synergistic activity of these two drugs on U-937 and on cultured cells from a leukemic patient. Proliferation was studied by 3H-thymidine incorporation; differentiation markers were evaluated immunologically by monoclonal antibodies and by cytochemical tests. Phagocytosis and NBT reduction test were also performed in order to confirm the differentiating properties of these drugs. Finally, the expression of the 1,25(OH)2D3 receptor was evaluated by immunochemical methods. After culturing these cells for 72 hours in the presence of 1,25(OH)2D3, cell proliferation was reduced and the expression of some phenotypic and functional markers suggested monocytic-macrophagic differentiation. Alpha interferon and 1,25(OH)2D3 synergistically inhibit the proliferation of U-937 cells. Alpha interferon increased the expression of the 1,25(OH)2D3 receptor in U-937 cells. The reported results confirm the synergistic activity of INF and 1,25(OH)2D3 on cell proliferation in monoblastic cells. The possible role of the increased expression of the vitamin receptor in cells cultured in the presence of INF is discussed.


To ID or not to ID? Changes in classification rates of intellectual disability using DSM-5.
The Diagnostic and Statistical Manual of Mental Disorders-Fifth Edition (DSM-5) diagnostic criteria for intellectual disability (ID) include a change to the definition of adaptive impairment. New criteria require impairment in one adaptive domain rather than two or more skill areas. The authors examined the diagnostic implications of using a popular adaptive skill inventory, the Adaptive Behavior Assessment System-Second Edition, with 884 clinically referred children (ages 6-16). One hundred sixty-six children met DSM-IV-TR criteria for ID; significantly fewer (n = 151, p = .001) met ID criteria under DSM-5 (9% decrease). Implementation of DSM-5 criteria for ID may substantively change the rate of ID diagnosis. These findings highlight the need for a combination of psychometric assessment and clinical judgment when implementing the adaptive deficits component of the DSM-5 criteria for ID diagnosis.


Causes and risk factors for infant mortality in Nunavut, Canada 1999-2011.
The northern territory Nunavut has Canada's largest jurisdictional land mass with 33,322 inhabitants, of which 85% self-identify as Inuit. Nunavut has rates of infant mortality, postneonatal mortality and hospitalisation of infants for respiratory infections that greatly exceed those for the rest of Canada. The infant mortality rate in Nunavut is 3 times the national average, and twice that of the neighbouring territory, the Northwest Territories. Nunavut has the largest Inuit population in Canada, a population which has been identified as having high rates of Sudden Infant Death Syndrome (SIDS) and infant deaths due to infections. To determine the causes and potential risk factors of infant mortality in Nunavut, we reviewed all infant deaths (<1 yr) documented by the Nunavut Chief Coroner's Office and the Nunavut Bureau of Statistics (n=117; 1999-2011). Rates were compared to published data for Canada. Sudden death in infancy (SIDS/SUDI; 48%) and infection (21%) were the leading causes of infant death, with rates significantly higher than for Canada (2003-2007). Of SIDS/SUDI cases with information on sleep position (n=42) and bed-sharing (n=47), 29 (69%) were sleeping non-supine and 33 (70%) were bed-sharing. Of those bed-sharing, 23 (70%) had two or more additional risk factors present, usually non-supine sleep position. CPT1A P479L homozygosity, which has been previously associated with infant mortality in Alaska Native and British Columbia First Nations populations, was associated with unexpected infant death (SIDS/SUDI, infection) throughout Nunavut (OR:3.43, 95% CI:1.30-11.47). Unexpected infant deaths comprise the majority of infant deaths in Nunavut. Although the CPT1A P479L variant was associated with unexpected infant death in Nunavut as a whole, the association was less apparent when population stratification was considered. Strategies to promote safe sleep practices and further understand other potential risk factors for infant mortality (P479L variant, respiratory illness) are underway with local partners.


The genes encoding virulence-associated proteins and the capsule of Streptococcus pneumoniae are upregulated and differentially expressed in vivo.
The polysaccharide capsule of Streptococcus pneumoniae and several well-characterized virulence proteins are known to contribute to the pathogenesis of pneumococcal disease. However, there is a paucity of data on the expression of their respective genes in vivo. In this study, the relative abundance of the mRNA transcripts of the genes encoding pneumolysin (ply), pneumococcal surface protein A (pspA), pneumococcal surface antigen A (psaA) and choline-binding protein A (cbpA), and of the first gene of the capsular polysaccharide biosynthesis locus (cps2A), was measured in virulent type 2 pneumococci harvested from the blood of BALB/c mice at 12 h and 24 h following intraperitoneal infection. The mRNA levels were then compared, using relative quantitative RT-PCR, with those present in organisms grown in serum broth. The expression of ply was upregulated threefold at 12 h, and 10-fold at 24 h post-infection; the expression of pspA and psaA was upregulated threefold and fivefold, respectively, at 12 h post-infection. Interestingly, the expression of pspA was 36-fold higher at 24 h post-infection whereas the expression of cps2A was upregulated approximately fourfold at 12 and 24 h post-infection. However, cbpA mRNA levels remained comparable in vivo and in vitro. When organisms were grown in whole blood or THY broth, the relative expression of these genes in the two growth media also differed markedly. This work provides direct molecular evidence that known virulence-associated genes of S. pneumoniae are differentially expressed in vivo. Data on the relative expression of these genes in different growth media also suggests that the regulation of expression of these genes is highly complex and multifactorial.


Enhanced expression of membrane type-1 matrix metalloproteinase in mesangial proliferative glomerulonephritis.
Matrix metalloproteinase-2 (MMP-2, gelatinase A) is involved in the inflammatory and sclerotic events of glomerular diseases. Newly identified membrane-type matrix metalloproteinases (MT-MMP) have been shown to activate specifically proMMP-2. To date, several types of MT-MMP have been cloned; however, their expressions in glomerular diseases have not been evaluated. To investigate the role of MT-MMP in glomerular diseases, the glomerular gene expression and enzymatic activity of MT-MMP were examined during the time course of nephritis induced in rats by anti-Thy1.1 antibody injection. Both MT1-MMP and MMP-2 mRNA expression increased prominently 5 and 10 d after anti-Thy1.1 antibody injection and decreased thereafter, as assayed by semiquantitative reverse transcription-PCR. In contrast, there were no remarkable changes in the gene expression of MT2-MMP between normal and diseased tissue, and that of MT3-MMP was not detected in isolated glomeruli by reverse transcription-PCR analysis. The activation of proMMP-2 as analyzed by gelatin zymography correlated with the glomerular MT1-MMP gene expression, suggesting that proMMP-2 was activated by MT1-MMP. Protein and mRNA expression of fibronectin, one of the major mesangial matrix proteins and substrate of MMP-2, were also synchronized with MT1-MMP and MMP-2 expression. In situ hybridization revealed intense MT1-MMP mRNA expression in the proliferating mesangial cells. Interestingly, MT1-MMP gene expression exhibited a similar distribution as alpha-smooth muscle actin expression, which was closely associated with mesangial phenotypic change. These results suggest that among the newly identified MT-MMP, MT1-MMP may play the central role in activation of proMMP-2. Furthermore, the enhancement of MT1-MMP and MMP-2 expression associated with mesangial phenotypic change may contribute to the development of anti-Thy1.1 antibody-induced glomerulonephritis and remodeling of extracellular matrices.


Engineering mammalian cells to seek senescence-associated secretory phenotypes.
Since the removal of senescent cells in model organisms has been linked to rejuvenation and increased lifespan, senotherapies have emerged to target senescent cells for death. In particular, interleukin-6 (IL6) is a prominent senescence-associated secretory phenotype (SASP) and, thus, seeking IL6 could potentially localize engineered cells to senescent cells for therapeutic intervention. Here, we engineered a chimeric IL6 receptor (IL6Rchi) that generates a Ca2+ signal in response to IL6 stimulation. When IL6Rchi was co-expressed with an engineered Ca2+-activated RhoA (CaRQ), it enabled directed migration to IL6 in cells that have no such natural ability. Next, the removal of target cells was accomplished by the mechanism of membrane fusion and subsequent death. This work represents a first step towards engineering a cell to target senescent cells that secrete high levels of IL6. For increased specificity to senescent cells, it will likely be necessary for an engineered cell to recognize multiple SASPs simultaneously.


Metabolism of the isoflavones genistein and biochanin A in human breast cancer cell lines.
There is substantial variation in the growth inhibition of different human breast cancer cell lines by the isoflavones genistein and biochanin A. ZR-75-1 and BT-20 cells are > or = 2- to 4-fold less sensitive to these isoflavones than are MCF-7 cells, whereas T47D cells have a sensitivity similar to that of MCF-7 cells. To determine whether these differences are related to isoflavone metabolism by these cancer cells, each of the cell lines was incubated with [4-(14)C]genistein and [4-(14)C]biochanin A. Metabolites in the cell culture media were identified by radio-HPLC electrospray ionization mass spectrometry. One metabolite of genistein (genistein 7-sulfate) and 2 metabolites of biochanin A (genistein and genistein 7-sulfate) were detected by radio-HPLC. Further analysis by mass spectrometry identified 3 other metabolites, a hydroxylated methylated form of each isoflavone and a biochanin A sulfate. IC50 (the concentration at which the growth rate was halved) values of the breast cancer cell lines did not correlate well with production of genistein 7-sulfate from genistein or with biochanin A sulfate, genistein 7-sulfate, or genistein from biochanin A. However, IC50 values correlated with the production of the hydroxylated and methylated forms of the isoflavones. Only T47D cells produced these metabolites in this study, and only T47D cells had IC50 values similar to those of MCF-7 cells, which also produced the hydroxylated and methylated metabolites. These data suggest that the hydroxylated and methylated metabolites may be the active forms of genistein in human breast cancer cells and emphasize the importance of isoflavone metabolism in the mechanism of action of isoflavones.


Value of Packaged Testing for Sexually Transmitted Infections for Persons who Inject Drugs Hospitalized With Serious Injection-Related Infections.
Persons who inject drugs (PWID) are frequently admitted for serious injection-related infections (SIRIs). PWID are also at risk for sexually transmitted infections (STIs). We conducted a multicenter quality improvement project at 3 hospitals in Missouri. PWID with SIRI who received an infectious diseases consultation were prospectively identified and placed into an electronic database as part of a Centers for Disease Control and Prevention-funded quality improvement project. Baseline data were collected from 8/1/2019 to 1/30/2020. During the intervention period (2/1/2020-2/28/2021), infectious diseases physicians caring for patients received 2 interventions: (1) email reminders of best practice screening for HIV, viral hepatitis, and STIs; (2) access to a customized EPIC SmartPhrase that included checkboxes of orders to include in assessment and plan of consultation notes. STI screening rates were compared before and after the intervention. We then calculated odds ratios to evaluate for risk factors for STIs in the cohort. Three hundred ninety-four unique patients were included in the cohort. Initial screening rates were highest for hepatitis C (88%), followed by HIV (86%). The bundled intervention improved screening rates for all conditions and substantially improved screening rates for gonorrhea, chlamydia, and syphilis (30% vs 51%, 30% vs 51%, and 39 vs 60%, respectively; P < .001). Of patients who underwent screening, 16.9% were positive for at least 1 STI. In general, demographics were not strongly associated with STIs. PWID admitted for SIRI frequently have unrecognized STIs. Our bundled intervention improved STI screening rates, but additional interventions are needed to optimize screening.


Effects of gamma irradiation on ruminal DM and NDF degradation kinetics of alfalfa hay.
The effects of gamma irradiation on ruminal dry matter, Neutral Detergent Fiber (NDF) degradation of alfalfa hay were investigated. Alfalfa hay samples were irradiated by gamma irradiator at doses of 50, 100 and 150 kGy under identical conditions of temperature and humidity. Nylon bags of untreated or irradiated samples were suspended in the rumen of three Taleshi bulls for up to 96 h and resulting data were fitted to non-linear degradation model to calculate degradation parameters. Results indicated that the washout fractions of dry matter and NDF increased linearly (p < 0.001) with increasing irradiation dose. The b fraction and the degradation rate of the b fraction (c) of DM and NDF were the highest at 50 kGy dose. Effective degradability of DM and NDF increased linearly with increasing irradiation dose. Gamma irradiation at doses of 50, 100 and 150 kGy increased the effective NDF degradability of alfalfa hay at rumen outflow rate of 0.05 h(-1) by about 8, 11 and 12%, respectively. Gamma irradiation affects on the hydrogenic bonds and with theirs breakdowning causes the wander-valls power weaken, that results in the degradation of cellulose and increasing of DM and NDF degradability.