Respiratory syncytial virus infection in the absence of STAT 1 results in airway dysfunction, airway mucus, and augmented IL-17 levels.
Respiratory syncytial virus (RSV) is the leading infectious cause of respiratory failure and wheezing in infants and young children. Prematurity is the greatest risk factor for severe RSV-induced disease, and recent studies suggest that premature children have lower levels of the type I IFNs (alpha/beta), for which signal transducer and activator of transcription (STAT) 1 is a critical intracellular signaling molecule. We hypothesized that RSV infection in STAT 1 knockout (STAT 1 KO) mice would result in both increased airway resistance and airway hyperresponsiveness. Wild-type (WT) and STAT 1 KO mice on a BALB/c background were either RSV or mock infected. Phenotypic response to infection was assessed by means of plethysmography, immunohistochemistry, and lung cytokine measurement. We found that STAT 1 KO mice infected with RSV (STAT 1 KO-RSV) had greater baseline lung resistance (P=.05) and airway responsiveness (P<.001) than mock-infected STAT 1 KO (STAT 1 KO-MOCK), RSV-infected wild type (WT-RSV), and mock-infected wild type (WT-MOCK) mice. In addition, the STAT 1 KO-RSV mice showed induction of mucus production and expression of gob-5 and Muc5ac, conditions not present in any of the other 3 groups. IL-17, a cytokine that regulates Muc5ac expression, was expressed in the lungs of the STAT 1 KO-RSV mice, whereas lung levels of IL-17 were undetectable in the remaining groups. Expression of the IL-23-specific p19 subunit was also increased in the STAT 1 KO-RSV mice but not in the WT-RSV mice. These results show that STAT 1 has an important regulatory role in RSV-induced alteration of airway function.


Impact of weather changes on air quality and related mortality in Spain over a 25 year period [1993-2017].
Climate change is a major public health concern. In addition to its direct impacts on temperature patterns and extreme weather events, climate change affects public health indirectly through its influence on air quality. Pollution trends are not only affected by emissions changes but also by weather changes. In this paper we analyze air quality trends in Spain of important air pollutants (C6H6, CO, NO2, NOx, O3, PM10, PM2.5, and SO2) recorded during the last 25 years, from 1993 to 2017. We found substantial reductions in ambient concentration levels for all the pollutants studied except for O3. To assess the influence of recent weather changes on air quality trends we applied generalized additive models (GAMs) using nonparametric smoothing; with and without adjusting for weather parameters including temperature, wind speed, humidity and precipitation frequency. The difference of annual slopes estimated by the models without and with adjusting for these meteorological variables represents the impact of weather changes on pollutant trends, i.e. the 'weather penalty'. The analyses were seasonally and geographically stratified to account for temporal and regional differences across Spain. The results were meta-analyzed to estimate weather penalties on ambient concentration trends at a national level as well as the impact on mortality for the most relevant pollutants. We found significant penalties for most pollutants, implying that air quality would have improved even more during our study period if weather conditions had remained constant. The largest weather influences were found for PM10, with seasonal penalties up to 22 μg⋅m-3 accumulated over the 25-year period in some regions. The national meta-analysis shows penalties of 0.060 μg⋅m-3 per year (95% Confidence Interval, CI: 0.004, 0.116) in cold months and 0.127 μg⋅m-3 per year (95% CI: 0.089, 0.164) in warm months. Penalties of this magnitude would correspond to 129 annual deaths (95% CI: 25, 233), i.e. approximately 3200 deaths over the 25-year period in Spain. According to our results, the health benefits of recent emission abatements for this pollutant in Spain would have been up to 10% greater if weather conditions had remained constant during the last 25 years.


Proteomic analysis of SETD6 interacting proteins.
SETD6 (SET-domain-containing protein 6) is a mono-methyltransferase that has been shown to methylate RelA and H2AZ. Using a proteomic approach we recently identified several new SETD6 substrates. To identify novel SETD6 interacting proteins, SETD6 was immunoprecipitated (IP) from Human erythromyeloblastoid leukemia K562 cells. SETD6 binding proteins were subjected to mass-spectrometry analysis resulting in 115 new SETD6 binding candidates. STRING database was used to map the SETD6 interactome network. Network enrichment analysis of biological processes with Gene Ontology (GO) database, identified three major groups; metabolic processes, muscle contraction and protein folding.


What is the most appropriate follow-up time for detecting the epidemiological relationship between coronary artery disease and its main risk factors: novel findings from a 35-year follow-up study.
The aim was to investigate the most appropriate follow-up time to detect the associations of coronary artery disease (CAD) with its traditional risk factors in a long-term prospective cohort study. The Kuopio Ischaemic Heart Disease Risk Factors Study provided the study material of 1958 middle-aged men free from CAD at baseline and followed up for 35 years. We performed Cox models adjusted for age, family history, diabetes, obesity, hypercholesterolemia, hypertension, smoking, and physical activity, investigated covariate interactions, and tested Schoenfeld residuals to detect time-dependent covariates. Moreover, we applied a sliding window procedure with a subarray of 5 years to better differentiate between risk factors manifested within years and those manifested within decades. The investigated manifestations were CAD and fatal acute myocardial infarction (AMI). Seven hundred seventeen (36.6%) men had CAD, and 109 (5.6%) men died from AMI. After 10 years of follow-up, diabetes became the strongest predictor of CAD with a fully adjusted hazard ratio (HR) of 2.5-2.8. During the first 5 years, smoking was the strongest predictor (HR 3.0-3.8). When the follow-up time was 8-19 years, hypercholesterolemia predicted CAD with a HR of >2. The associations of CAD with age and diabetes depended on time. Age hypertension was the only statistically significant covariate interaction. The sliding window procedure highlighted the significance of diabetes over the first 20 years and hypertension after that. Regarding AMI, smoking was associated with the highest fully adjusted HR (2.9-10.1) during the first 13 years. The associations of extreme and low physical activity with AMI peaked when the follow-up time was 3-8 years. Diabetes showed its highest HR (2.7-3.7) when the follow-up time was 10-20 years. During the last 16 years, hypertension was the strongest predictor of AMI (HR 3.1-6.4). The most appropriate follow-up time for most CAD risk factors was 10-20 years. Concerning smoking and hypertension shorter and longer follow-up times could be considered, respectively, particularly when studying fatal AMI. In general, prospective cohort studies of CAD would provide more comprehensive results by reporting point estimates in relation to more than one timepoint and concerning sliding windows.


Thrombin weakens the amnion extracellular matrix (ECM) directly rather than through protease activated receptors.
Preterm premature rupture of fetal membranes (pPROM) is a major cause of preterm birth. Abruption associated thrombin production, and infection-inflammation associated cytokine production reportedly play major roles in pPROM. Utilizing an in vitro model-system we have confirmed that both thrombin and inflammatory cytokines remodel and biomechanically weaken amnion, the load-bearing component of FM. Also, we have shown thrombin directly weakens isolated amnion but cytokines weaken amnion only indirectly by initially interacting with choriodecidua and releasing unidentified soluble activator(s). This study's purpose was to determine whether thrombin weakens the isolated amnion through thrombin receptor-protease activated receptors (PARs 1,2,3,4), activation of previously secreted extracellular matrix (ECM) enzymes, or by direct action on the ECM. Primary amnion cells and isolated amnion were tested for PARs by immunohistochemistry, Western Blot and rtPCR. Cell-free amnion ECM was produced by devitalizing isolated amnion by exposure to UV light and subsequent freeze-thaw cycles. Devitalized amnion membrane explants were incubated with thrombin and biomechanically tested. PARs were not found in amnion or amnion cells. Thrombin induced dose-dependent weakening of devitalized amnion explants. Preincubation with the thrombin inhibitor hirudin prevented thrombin-induced weakening. Thrombin converted pro-MMP2 embedded in the devitalized amnion ECM to multiple active forms. Thrombin also directly digested gelatin gels in zymograms suggesting the possibility of direct degradation of amnion ECM components. Thrombin appears to directly weaken the amnion ECM and additionally activates stored pro-MMP2 to active forms that may further enhance amnion ECM degradation. Thrombin weakens amnion directly rather than through PARs.


Aerobic Methanotrophy and Co-occurrence Networks of a Tropical Rainforest and Oil Palm Plantations in Malaysia.
Oil palm (OP) plantations are gradually replacing tropical rainforest in Malaysia, one of the largest palm oil producers globally. Conversion of lands to OP plantations has been associated with compositional shifts of the microbial community, with consequences on the greenhouse gas (GHG) emissions. While the impact of the change in land use has recently been investigated for microorganisms involved in N2O emission, the response of the aerobic methanotrophs to OP agriculture remains to be determined. Here, we monitored the bacterial community composition, focusing on the aerobic methanotrophs, in OP agricultural soils since 2012, 2006, and 1993, as well as in a tropical rainforest, in 2019 and 2020. High-affinity methane uptake was confirmed, showing significantly lower rates in the OP plantations than in the tropical rainforest, but values increased with continuous OP agriculture. The bacterial, including the methanotrophic community composition, was modified with ongoing OP agriculture. The methanotrophic community composition was predominantly composed of unclassified methanotrophs, with the canonical (Methylocystis) and putative methanotrophs thought to catalyze high-affinity methane oxidation present at higher relative abundance in the oldest OP plantation. Results suggest that the methanotrophic community was relatively more stable within each site, exhibiting less temporal variations than the total bacterial community. Uncharacteristically, a 16S rRNA gene-based co-occurrence network analysis revealed a more complex and connected community in the OP agricultural soil, which may influence the resilience of the bacterial community to disturbances. Overall, we provide a first insight into the ecology and role of the aerobic methanotrophs as a methane sink in OP agricultural soils.


Influencing and moderating factors analyzed in the group art therapy of two schizophrenic inpatients.
Art therapy has been considered a guideline treatment for schizophrenia. Due to difficulty in the outcome measurement, the research is difficult and controversial. Here, we presented two schizophrenic patients receiving the regular art group therapy. We compared their characteristics and different outcome. Art therapy is difficult to quantify. However, we could qualify the improvement from the individual case. Further study might be focus on how to make appropriate qualification of art therapy and individualized difference instead of enrollment of huge data bank.


"Hyperglutamatergic cortico-striato-thalamo-cortical circuit" breaker drugs alleviate tics in a transgenic circuit model of Tourette׳s syndrome.
The brain circuits underlying tics in Tourette׳s syndrome (TS) are unknown but thought to involve cortico/amygdalo-striato-thalamo-cortical (CSTC) loop hyperactivity. We previously engineered a transgenic mouse "circuit model" of TS by expressing an artificial neuropotentiating transgene (encoding the cAMP-elevating, intracellular A1 subunit of cholera toxin) within a small population of dopamine D1 receptor-expressing somatosensory cortical and limbic neurons that hyperactivate cortico/amygdalostriatal glutamatergic output circuits thought to be hyperactive in TS and comorbid obsessive-compulsive (OC) disorders. As in TS, these D1CT-7 ("Ticcy") transgenic mice׳s tics were alleviated by the TS drugs clonidine and dopamine D2 receptor antagonists; and their chronic glutamate-excited striatal motor output was unbalanced toward hyperactivity of the motoric direct pathway and inactivity of the cataleptic indirect pathway. Here we have examined whether these mice׳s tics are countered by drugs that "break" sequential elements of their hyperactive cortical/amygdalar glutamatergic and efferent striatal circuit: anti-serotonoceptive and anti-noradrenoceptive corticostriatal glutamate output blockers (the serotonin 5-HT2a,c receptor antagonist ritanserin and the NE alpha-1 receptor antagonist prazosin); agmatinergic striatothalamic GABA output blockers (the presynaptic agmatine/imidazoline I1 receptor agonist moxonidine); and nigrostriatal dopamine output blockers (the presynaptic D2 receptor agonist bromocriptine). Each drug class alleviates tics in the Ticcy mice, suggesting a hyperglutamatergic CSTC "tic circuit" could exist in TS wherein cortical/amygdalar pyramidal projection neurons׳ glutamatergic overexcitation of both striatal output neurons and nigrostriatal dopaminergic modulatory neurons unbalances their circuit integration to excite striatothalamic output and create tics, and illuminating new TS drug strategies.


Nutritional support in HIV infection and AIDS.
Weight loss and wasting are common features of HIV infection and AIDS. Patterns of weight loss can be acute or chronic which appear to be related primarily to systemic infections and gastrointestinal pathology, respectively. However, weight loss is not inevitable. and periods of weight stability and weight gain have been documented. Reduced food intake appears to be a major cause of weight loss in HIV infection. Since time of death has been associated with degree of wasting, it seems reasonable to suggest that nutritional support may contribute to enhancing survival and quality of life. All patients should have early access to a qualified dietitian such that assessment of individual situations can be made and appropriate dietary advice given, within a multi-disciplinary approach. Choice of nutritional therapy should be made based on an assessment of the causes of weight loss and an assessment of gut function. Treating infections and alleviating symptoms is vital for ensuring effective nutritional support. Enhancing the energy and protein density of foods and use of oral supplements should be considered if a normal diet alone cannot meet nutritional requirements. Unnecessary dietary restrictions should be avoided. Enteral feeding is indicated for patients unable to meet their needs via the oral route, and in cases of inadequate gut function, parenteral nutrition may be necessary. There is currently insufficient clinical evidence to justify the need for special enteral formulae specifically for patients with HIV infection.


Inorganic cobalt supplementation: prediction of cobalt levels in whole blood and urine using a biokinetic model.
Soluble cobalt (Co) supplements with recommended daily doses up to 1000 μg Co/day are increasingly being marketed to consumers interested in healthy living practices. For example, some athletes may consider using Co supplements as blood doping agents, as Co is known to stimulate erythropoesis. However, the distribution and excretion kinetics of ingested Co are understood in a limited fashion. We used a Co-specific biokinetic model to estimate whole blood and urine Co levels resulting from oral exposure or ingestion of Co in amounts exceeding typical dietary intake rates. Following 10 days of Co supplementation at a rate of 400 to 1000 μg/day, predicted adult Co concentrations range from 1.7 to 10 μg/L in whole blood, and from 20 to 120 μg/L in urine. Chronic supplementation (≥ 1 year) at a rate of 1000 μg Co/day is predicted to result in blood levels of 5.7 to 13 μg/L, and in urine levels from 65 to 150 μg/L. The model predictions are within those measured in humans following ingestion of known doses. The methodology presented in this paper can be used to predict urinary or blood Co levels following acute or chronic occupational incidental ingestion, medicinal therapy, supplemental intake, or other non-occupational exposures.


Teaching and preschoolers' ability to infer knowledge from mistakes.
For instruction to be effective, teachers must adjust the way they teach to match what learners know. We asked whether children's ability to infer what someone knows based on his or her mistakes develops alongside their teaching-children's use of more explicit teaching strategies and their ability to tailor how much information to provide in response to their pupils' mistakes. Preschoolers (N=48) were taught a simple game and were then introduced to four puppets: one puppet who played the game perfectly, two puppets who each made one mistake, and one puppet who made two mistakes. After watching each puppet play individually, children were asked to rate the puppet's understanding of the game and then were invited to teach the puppet. Children's ability to monitor the relative accuracy of the puppets-the ability to make nuanced judgments about what each puppet understood based on each puppet's unique mistakes-improved with age. Moreover, older children were more explicit and more precise teachers than younger children. They more often contrasted the learners' mistakes with what should be done and more often provided instructions that directly addressed the puppets' unique mistakes. Thus, between 3 and 5 years of age, developments in children's ability to infer knowledge from mistakes parallel developments both in the strategies children use to teach and in the amount of information they teach in response to mistakes.


Antibody-dependent cell lysis by NK cells is preserved after sarcoma-induced inhibition of NK cell cytotoxicity.
Osteosarcoma and Ewing's sarcoma tumor cells are susceptible to IL15-induced or antibody-mediated cytolytic activity of NK cells in short-term cytotoxicity assays. When encountering the tumor environment in vivo, NK cells may be in contact with tumor cells for a prolonged time period. We explored whether a prolonged interaction with sarcoma cells can modulate the activation and cytotoxic activity of NK cells. The 40 h coculture of NK cells with sarcoma cells reversibly interfered with the IL15-induced expression of NKG2D, DNAM-1 and NKp30 and inhibited the cytolytic activity of NK cells. The inhibitory effects on receptor expression required physical contact between NK cells and sarcoma cells and were independent of TGF-β. Five days pre-incubation of NK cells with IL15 prevented the down-regulation of NKG2D and cytolytic activity in subsequent cocultures with sarcoma cells. NK cell FcγRIIIa/CD16 receptor expression and antibody-mediated cytotoxicity were not affected after the coculture. Inhibition of NK cell cytotoxicity was directly linked to the down-regulation of the respective NK cell-activating receptors. Our data demonstrate that the inhibitory effects of sarcoma cells on the cytolytic activity of NK cells do not affect the antibody-dependent cytotoxicity and can be prevented by pre-activation of NK cells with IL15. Thus, the combination of cytokine-activated NK cells and monoclonal antibody therapy may be required to improve tumor targeting and NK cell functionality in the tumor environment.


Continence in patients who undergo posterior rectal flap anorectoplasty.
The authors report the postoperative follow-up results of patients who underwent posterior rectal flap anorectoplasty (PRFA), with emphasis in fecal continence. Variables considered on each patient were sex, presence of fistula, location of the fistula, severity of the defect during the correction, sacral condition, dysraphism, and age at surgery. Continence was rated using the Peña method and defecogram. The authors studied 20 patients (17 boys and three girls). Postoperatively, normal continence was found in 18 patients. Of the two remaining (boys), one had grade II spotting (sacrum agenesia and uretrobulbar fistula) and the other had grade II staining; this patient had the most severe malformation requiring an abdominoperineal approach. All of the defecograms showed complete emptying of the rectum. No recurrent fistulas occurred. The posterior rectal flap anorectoplasty is a new technique that takes into consideration all known factors that contribute to continence. This initial report confirms that this technique results in excellent continence for patients with imperforate anus.


Lower embryonic loss rates among twin gestations following assisted reproduction.
To determine whether maternal age and number of transferred embryos influence early pregnancy losses in twin pregnancies compared to singletons following IVF/ICSI. We compared the pregnancy loss rates in singleton (n = 549) and twin (n = 252) gestations, stratified by maternal age (< or = 35 and > 35 years) and the number of transferred embryos (1-3 and 4-9). Loss rates of singleton pregnancies were significantly higher than that in twins (OR 3.0, 95% CI 1.9, 4.9), especially among singletons conceived after transfer of 4-9 embryos (OR 5.0, 95% CI 2.2, 11.9). Younger mothers of twins had lower loss rates (OR 0.3, 95% CI 0.1, 0.9). Twins have a significantly reduced spontaneous miscarriage rate compared with singletons following IVF/ICSI. Higher implantation rates per cycle (i.e., development of twins rather than one live embryo) may represent a better capacity of the uterus for early embryonic development.


Predictors for Unfavorable Early Outcomes in Elective Total Hip Arthroplasty: Does Extreme Body Mass Index Matter?
Studies of previous cohorts have demonstrated a controversial association between extreme body mass index (BMI) and complication rates following total hip arthroplasty (THA). The purpose of this study was to compare 30-day perioperative complications in underweight (BMI <18.50 kg/m2), normal-weight (BMI 18.50-24.99 kg/m2), overweight (BMI 25.00-29.99 kg/m2), class I obesity (BMI 30.00-34.99 kg/m2), and morbidly obese (BMI ≥35.00 kg/m2) groups. We performed a cohort study including patients who underwent unilateral primary THA by a single surgeon between January 2010 and December 2015 at our institution. We assessed 30-day complications, operation time, operative blood loss, and length of hospital stay. We identified 1565 primary THAs that were performed in patients with varying BMI levels. Compared with the normal-weight patients, the morbidly obese group had a higher 30-day complication rate (8.9% vs. 2.4%), longer operative time (79 minutes vs. 70 minutes), and more blood loss (376 mL vs. 302 mL). Underweight patients did not present any 30-day complications, and there were no differences among underweight and normal-weight patients regarding complication rates, operative time, or blood loss. The mean length of hospital stay was comparable among the different BMI groups. In the multivariate regression model, higher BMI was not associated with a higher risk of 30-day complications. Independent risk factors for 30-day complications were advanced age, prolonged operative time, and cardiovascular comorbidities. Although increased operative time, blood loss, and perioperative complications were seen in the morbidly obese patients, BMI alone was not an independent risk factor for a higher 30-day complication rate. Therefore, our data suggest clinicians should make elderly patients aware of increased 30-day complications before the procedure, particularly those with cardiovascular comorbidities. Withholding THA solely on the basis of BMI is not justified.


Leptin Induces a Novel Form of NMDA Receptor-Dependent LTP at Hippocampal Temporoammonic-CA1 Synapses
It is well documented that the hormone leptin regulates many central functions and that hippocampal CA1 pyramidal neurons are a key target for leptin action. Indeed, leptin modulates excitatory synaptic transmission and synaptic plasticity at the Schaffer-collateral input to CA1 neurons. However the impact of leptin on the direct temporoammonic (TA) input to CA1 neurons is not known. Here we show that leptin evokes a long-lasting increase [long-term potentiation (LTP)] in excitatory synaptic transmission at TA-CA1 synapses in rat juvenile hippocampus. Leptin-induced LTP was NMDA receptor-dependent and specifically involved the activation of GluN2B subunits. The signaling pathways underlying leptin-induced LTP involve the activation of phosphoinositide 3-kinase, but were independent of the ERK signaling cascade. Moreover, insertion of GluA2-lacking AMPA receptors was required for leptin-induced LTP as prior application of philanthotoxin prevented the effects of leptin. In addition, synaptic-induced LTP occluded the persistent increase in synaptic efficacy induced by leptin. In conclusion, these data indicate that leptin induces a novel form of NMDA receptor-dependent LTP at juvenile TA-CA1 synapses, which has important implications for the role of leptin in modulating hippocampal synaptic function in health and disease.


The functional role of the Meis/Prep-binding elements in Pax6 locus during pancreas and eye development.
Pax6 is an essential transcription factor for lens, lacrimal gland and pancreas development. Previous transgenic analyses have identified several Pax6 regulatory elements, but their functional significance and binding factors remain largely unknown. In this study, we generated two genomic truncations to delete three elements that were previously shown to bind to the Meis/Prep family homeoproteins. One 3.1 kb deletion (Pax6(∆DP/∆DP)) removed two putative pancreatic enhancers and a previously identified ectodermal enhancer, while a 450 bp sub-deletion (Pax6(∆PE/∆PE)) eliminated only the promoter-proximal pancreatic enhancer. Immunohistochemistry and quantitative RT-PCR showed that the Pax6(∆PE/∆PE) pancreata had a significant decrease in Pax6, glucagon, and insulin expression, while no further reductions were observed in the Pax6(∆DP/∆DP) mice, indicating that only the 450 bp region is required for pancreatic development. In contrast, Pax6(∆DP/∆DP), but not Pax6(∆PE/∆PE) mice, developed stunted lacrimal gland and lens hypoplasia which was significantly more severe than that reported when only the ectodermal enhancer was deleted. This result suggested that the ectodermal enhancer must cooperate with its neighboring sequences to regulate the Pax6 ectodermal expression. Finally, we generated conditional knockouts of Prep1 in the lens and pancreas, but surprisingly, did not observe any developmental defects. Together, these results provide functional evidence for the independent and synergistic roles of the Pax6 upstream enhancers, and they suggest the potential redundancy of Meis/Prep protein in Pax6 regulation.


Association of Depression With Past-Month Cannabis Use Among US Adults Aged 20 to 59 Years, 2005 to 2016.
Despite studies showing that repeated cannabis use may worsen depressive symptoms, the popular media increasingly presents cannabis as beneficial to mental health, and many members of the public view cannabis as beneficial for depression. Therefore, cannabis use among individuals with depression may be becoming more prevalent. To examine the association of depression with past-month cannabis use among US adults and the time trends for this association from 2005 to 2016. This repeated cross-sectional study used data from 16 216 adults aged 20 to 59 years who were surveyed by the National Health and Nutrition Examination Survey, a national, annual, cross-sectional survey in the United States, between 2005 and 2016. Data analysis was conducted from January to February 2020. Survey year and depression, as indicated by a score of at least 10 on the Patient Health Questionnaire-9. Any past-month cannabis use (ie, ≥1 use in the past 30 days) and daily or near-daily past-month cannabis use (ie, ≥20 uses in the past 30 days). Logistic regression was used to examine time trends in the prevalence of cannabis use, depression, and the association between cannabis use and depression from 2005 to 2016. The final analysis included 16 216 adults, of whom 7768 (weighted percentage, 48.9%) were men, 6809 (weighted percentage, 66.4%) were non-Hispanic White participants, and 9494 (weighted percentage, 65.6%) had at least some college education. They had a weighted mean age of 39.12 (95% CI, 38.23-39.40) years. Individuals with depression had 1.90 (95% CI, 1.62-2.24) times the odds of any past-month cannabis use and 2.29 (95% CI, 1.80-2.92) times the odds of daily or near-daily cannabis use compared with those without depression. The association between cannabis use and depression increased significantly from 2005 to 2016. The odds ratio for depression and any past-month cannabis use increased from 1.46 (95% CI, 1.07-1.99) in 2005 to 2006 to 2.30 (95% CI, 1.82-2.91) in 2015 to 2016. The odds ratio for depression and daily or near-daily past-month cannabis use increased from 1.37 (95% CI, 0.81-2.32) in 2005 to 2006 to 3.16 (95% CI, 2.23-4.48) in 2015 to 2016. The findings of this study indicate that individuals with depression are at increasing risk of cannabis use, with a particularly strong increase in daily or near-daily cannabis use. Clinicians should be aware of these trends and the evidence that cannabis does not treat depression effectively when discussing cannabis use with patients.


Data on length-weight and length-length relationships, mean condition factor, and gonadosomatic index of Rutilus rutilus and Perca fluviatilis from the Ob River basin, Western Siberia.
Raw data were obtained from 250 specimens of roach Rutilus rutilus and 274 specimens of perch Perca fluviatilis at 15 sampling sites located from North to South Taiga. Fish sampling was performed using gill-nets with 22 and 28 mm mesh. Total and eviscerated weights, total and standard lengths, fish sex, gonad weight and maturity stage were recorded. Linear regression analysis of log-transformed total weight and total length values was performed. Regression slope and intercept were used to obtain length-weight relationships and mean condition factor values. Length-length relationships were calculated by the linear regression between the total and standard lengths. Gonadosomatic index was determined using the gonad weight and the total weight of fish. The data are useful for establishing biomass, fish growth patterns, relative condition of individuals within a sample or across populations.


Therapeutic monitoring of metipamide during antihypertensive therapy.
The aim of our study was to monitor metipamide during a two-month period of treatment and to determine whether the whole-blood levels estimated by high-performance liquid chromatography provide a relevant indicator of possible accumulation of the drug. We also analysed antihypertensive activity and biochemical changes in the blood of twenty hypertonic patients. The results of our clinical trial showed that metipamide is an effective first-line antihypertensive agent, in that it combines satisfactory reduction of blood pressure with a low frequency of side-effects and a simple once-daily dosage regime.


Candidate gene based SSR and SNP markers for gynoecy in bitter gourd (Momordica charantia L.).
Even though the bitter gourd hybrids are shown to have significant heterosis for many of the economic traits, processes such as manual bagging and hand pollination make the hybrid seed production labour-intensive. Use of gynoecious line as female parent makes hybrid seed production more economical. This work was performed with the objective to identify the candidate gene based molecular markers for gynoecy in bitter gourd. Seven putative genes for flowering and sex expression, isolated from the monoecious (MC-136) and gynoecious (KAU-MCGy-101) bitter gourd accessions, were sequence characterized. MADS-box transcription factor genes AG6 and McAG2 had nucleotide polymorphisms at five sites each and were potential candidates for marker development. An In/Del polymorphism of 48 bp ([TC]24) in AG6 gene was used to develop an SSR marker and a transition mutation of [A/G] in this gene was used to develop a set of SNP markers. These markers have developed distinct polymorphism between the monoecious and gynoecious genotypes and were found suited for the marker assisted selection. MADS box transcription factor genes AG6 and McAG2 are identified as candidates for sex expression in bitter gourd. Based on the InDels and transition in the intronic region of AG6, SSR marker BGAG6 and an SNP marker set segregating with the sex forms were developed. The markers have been validated using four other monoecious lines and are routinely used in our bitter gourd hybrid seed production programmes.


Immune checkpoint inhibitor monotherapy is associated with less cardiac toxicity than combination therapy.
Treatment options for many cancers include immune checkpoint inhibitor (ICI) monotherapy and combination therapy with impressive clinical benefit across cancers. We sought to define the comparative cardiac risks of ICI combination and monotherapy. We used VigiBase, the World Health Organization pharmacovigilance database, to identify cardiac ADRs (cADRs), such as carditis, heart failure, arrhythmia, myocardial infarction, and valvular dysfunction, related to ICI therapy. To explore possible relationships, we used the reporting odds ratio (ROR) as a proxy of relative risk. A lower bound of a 95% confidence interval of ROR > 1 reflects a disproportionality signal that more ADRs are observed than expected due to chance. We found 2278 cADR for ICI monotherapy and 353 for ICI combination therapy. Combination therapy was associated with significantly higher odds of carditis (ROR 6.9, 95% CI: 5.6-8.3) versus ICI monotherapy (ROR 5.0, 95% CI: 4.6-5.4). Carditis in ICI combination therapy was fatal in 23.4% of reported ADRs, compared to 15.8% for ICI monotherapy (P = 0.058). Using validated pharmacovigilance methodology, we found increased odds of carditis for all ICI therapies, with the highest odds for combination therapy. Given the substantial risk of severe ADR and death, clinicians should consider these findings when prescribing checkpoint inhibitors.


Digital EPR with an arbitrary waveform generator and direct detection at the carrier frequency.
A digital EPR spectrometer was constructed by replacing the traditional bridge with an arbitrary waveform generator (AWG) to produce excitation patterns and a high-speed digitizer for direct detection of the spin system response at the carrier frequency. Digital down-conversion produced baseband signals in quadrature with very precise orthogonality. Real-time resonator tuning was performed by monitoring the Fourier transforms of signals reflected from the resonator during frequency sweeps generated by the AWG. The capabilities of the system were demonstrated by rapid magnetic field scans at 256 MHz carrier frequency, and FID and spin echo experiments at 1 and 10 GHz carrier frequencies. For the rapid scan experiments the leakage through a cross-loop resonator was compensated by adjusting the amplitude and phase of a sinusoid at the carrier frequency that was generated with another AWG channel.


[Dermoid cysts of the floor of mouth].
After brief mention of the embryogenesis of oral organs, a case of congenital dermoid cyst of the floor of the mouth treated surgically intraorally is reported. A short discussion on differential diagnosis follows.


Effect of waste tire rubber crumb on impermeability of cement-stabilized soil.
To alleviate the application limitations of cement-stabilized soil in impermeable engineering and promote the recycling of waste rubber tires, crumb rubber produced from waste rubber tires was used to improve the engineering properties of the soil. The effect of crumb rubber on the permeability of the soil under different conditions was investigated using compression, compaction, and permeability tests. Crumb rubber can effectively improve the impermeability of cement-stabilized soil. The impermeability efficiency of crumb rubber is between 11% and 45%. Cleaning crumb rubber with water and Na2CO3 solution can reduce the hydraulic conductivity of rubberized cement-stabilized soil (RCSS) by 7.9%-63.6%. This, in turn, increases the unconfined compressive strength by 4.1%-25.5%. The hydraulic conductivity of the RCSS decreases with an increase in the cement content, curing duration, and void ratio. A crumb rubber concentration of 10%-20% is suitable for enhancing the impermeability of the RCSS and satisfying its strength requirements. The NOF of the equation used to predict the hydraulic conductivity of the RCSS by the rubber content, cement content, and curing duration is less than 0.35. The linear correlation between the predicted and measured values of hydraulic conductivity was determined to be 0.995 from the k-quc correlation model. The results show that the hydraulic conductivity of RCSS can be estimated reliably based on the mix ratio and compressive strength.


Integrating Machine Learning and Tumor Immune Signature to Predict Oncologic Outcomes in Resected Biliary Tract Cancer.
Improved methods are needed to predict outcomes in biliary tract cancers (BTCs). We aimed to build an immune-related signature and establish holistic models using machine learning. Samples were from 305 BTC patients treated with curative-intent resection, divided into derivation and validation cohorts in a two-to-one ratio. Spatial resolution of T cell infiltration and PD-1/PD-L1 expression was assessed by immunohistochemistry. An immune signature was constructed using classification and regression tree. Machine learning was applied to develop prediction models for disease-specific survival (DSS) and recurrence-free survival (RFS). The immune signature composed of CD3+, CD8+, and PD-1+ cell densities and PD-L1 expression within tumor epithelium significantly stratified patients into three clusters, with median DSS varying from 11.7 to 80.8 months and median RFS varying from 6.2 to 62.0 months. Gradient boosting machines (GBM) outperformed rival machine-learning algorithms and selected the same 11 covariates for DSS and RFS prediction: immune signature, tumor site, age, bilirubin, albumin, carcinoembryonic antigen, cancer antigen 19-9, tumor size, tumor differentiation, resection margin, and nodal metastasis. The clinical-immune GBM models accurately predicted DSS and RFS, with respective concordance index of 0.776-0.816 and 0.741-0.781. GBM models showed significantly improved performance compared with tumor-node-metastasis staging system. The immune signature promises to stratify prognosis and allocate treatment in resected BTC. The clinical-immune GBM models accurately predict recurrence and death from BTC following surgery.


Recent Hospitalization and Initiation of Antiepileptics Among Persons With Alzheimer's Disease.
Antiepileptic drugs (AEDs) are frequently prescribed for persons with Alzheimer's disease (AD), but little is known on factors associated with AED initiation in this population. We investigated whether recent hospitalization is associated with AED initiation in persons with AD. Nested case-control study in the nationwide register-based Medication use and Alzheimer's disease (MEDALZ) cohort. The MEDALZ cohort includes 70,718 persons diagnosed with AD during 2005-2011 in Finland. Altogether 6814 AED initiators and 6814 age-, sex-, and time since AD diagnosis-matched noninitiators were included in this study. Matching date was the date of AED initiation. AED purchases were identified from the Prescription Register and hospitalizations from the Care Register for Health Care. Recent hospitalization was defined as hospitalization ending within 2 weeks before the matching date. Association between recent hospitalization and AED initiation was assessed with conditional logistic regression. The most frequently initiated AEDs were pregabalin (42.9%) and valproic acid (32.2%). A bigger proportion of AED initiators (36.9%) than noninitiators (5.3%) were recently hospitalized [odds ratio (OR) 10.5, 95% CI 9.22-11.9]. Dementia was the most frequent discharge diagnosis among AED initiators (29.1%) and noninitiators (27.9%). Among AED initiators, the next most frequent diagnosis was epilepsy (20.6%). Musculoskeletal diagnoses and use of analgesics including opioids was more common among gabapentinoid initiators compared with other AED initiators. Recent hospitalization was significantly related to AED initiation. Initiations of AED might have been related to common symptoms in persons with AD like neuropathic pain, epilepsy, and neuropsychiatric symptoms.


Giant intracranial aneurysm presenting as a posterior fossa mass.
A patient with signs of brainstem compression and hydrocephalus is reported. The clinical picture suggested a posterior fossa neoplasm. Computed tomography, however, proved highly accurate in making the diagnosis of a giant, partially thrombosed vertebrobasilar aneurysm. The radiographic appearance of this entity is summarized and pathologic correlation is offered.


GPR84 in physiology-Many functions in many tissues.
Members of the GPCR superfamily have a wide variety of physiological roles and are therefore valuable targets for developing effective medicines. However, within this superfamily are receptors that are less well characterized and remain orphans, including GPR84. This receptor is stimulated by ligands derived from dietary nutrients, specifically medium chain fatty acids (C9-14), and novel synthetic agonists. There are data demonstrating the role of GPR84 in inflammatory pathways, in addition to emerging data suggesting a key role for GPR84 as a nutrient-sensing GPCR involved in metabolism by sensing energy load via nutrient exposure and subsequent signalling leading to modulation of food intake. Exploring GPR84 pharmacology, its localization and what drives its expression has revealed multiple roles for this receptor. Here, we will reflect on these various roles of GRP84 demonstrated thus far, primarily by exploring data from pre-clinical and clinical studies in various physiological systems, with a specific focus on the gastrointestinal tract.


Pharmacological targeting of microglia dynamics in Alzheimer's disease: Preclinical and clinical evidence.
Numerous clinical trials of anti-amyloid agents for Alzheimer's disease (AD) were so far unsuccessful thereby challenging the validity of the amyloid hypothesis. This lack of progress has encouraged researchers to investigate alternative mechanisms in non-neuronal cells, among which microglia represent nowadays an attractive target. Microglia play a key role in the developing brain and contribute to synaptic remodeling in the mature brain. On the other hand, the intimate relationship between microglia and synapses led to the so-called synaptic stripping hypothesis, a process in which microglia selectively remove synapses from injured neurons. Synaptic stripping, along with the induction of a microglia-mediated chronic neuroinflammatory environment, promote the progressive synaptic degeneration in AD. Therefore, targeting microglia may pave the way for a new disease modifying approach. This review provides an overview of the pathophysiological roles of the microglia cells in AD and describes putative targets for pharmacological intervention. It also provides evidence for microglia-targeted strategies in preclinical AD studies and in early clinical trials.


Regurgitation across an obviously normal valve orifice as a possible prerequisite for non-organic cardiac murmur.
Using pulsed and continuous wave Doppler echocardiography, the author examined 359 subjects referred to ultrasound examination for a variety of reasons. Thorough examination of persons with morphologically unchanged valves revealed regurgitation in the tricuspid, pulmonary, mitral and aortic valves in 49.4%, 20%, 14.5% and 0.8%, respectively. The degree of regurgitation was invariably haemodynamically insignificant. Some patients had a positive finding in several valves at a time. Overall, regurgitation was found in 247 subjects (69%). While it is evident from the study that the finding of regurgitation in normal valves is relatively frequent, evaluation of its clinical importance remains an open question.


Prevalence of sport specialisation and association with injury history in youth football.
The aims of this study were to determine the prevalence of sport specialisation in youth football, and to investigate the associations of sport specialisation and volume of sport participation with injury history. Cross-sectional questionnaire. New Zealand youth football teams. 414 youth football players aged 10-15 years. The level of specialisation, average weekly sport participation and free-play volume were recorded. 12-month injury history was captured and grouped by injury type. Associations between level of specialisation and demographic variables were analysed using chi square tests. Logistic regression was used to investigate associations between injury history, level of specialisation, and volume of participation. Participants were classified as high (43%), moderate (38%), or low (19%) specialised (n = 399 complete). High specialisation was more likely in boys, older participants, and those from large schools. Highly specialised participants were more likely to report a history of gradual onset injury than those who were low specialised (n = 340 with complete injury data). Odds of reporting a gradual onset injury also increased with greater weekly and annual sport participation volume. There is a high prevalence of sport specialisation in youth football, and it is associated with increased incidence of gradual onset injury.


Distortion products otoacoustic emissions in diagnosis of hearing loss in Down syndrome.
This study was carried out to investigate the features of hearing impairment in subjects with Down syndrome. Forty-seven children and 14 adults with Down syndrome were included in the study. In all cases, a complete otorhinolaryngological examination was performed, followed by audiological assessment. Depending on age, intellectual level and middle ear status the following examinations were performed: pure-tone 'play audiometry', tympanometry, acoustic reflex, auditory brain response (ABR) and distortion products otoacoustic emissions (DPOAE). The results were compared with age matched control groups. Among the group of children with Down syndrome, we have frequently found impairment of the conductive function of the middle ear which was expressed by pathological tympanometry. Tympanometry of B and C type was detected in 56% of ears. The amplitude of DPOAE was lower in children with Down syndrome than in the control group. This difference was more expressed in adults with Down syndrome. DPOAE examination results in subjects with Down syndrome without conductive hearing loss indicate early age related inner ear impairment.


A Study on Outcome of Myringoplasty in Dry Ear (Quiescent/Inactive CSOM) Without Using Gelfoam in Middle Ear.
Myringoplasty is a common surgery done for patients with tympanic membrane perforation in our clinical practice. This study was done to know the outcome of myringoplasty done without using gelfoam as a scaffold material. Simultaneously other parameters influencing the outcome were analysed. In a total of 80 patients assessed over 25 months were divided into test and control group by quasi randomisation. All patients underwent endoscopic underlay myringoplasty. Control groups had gelfoam bed created with gelfoam, whereas the test group only middle ear air pocket created with a proper seal without using gelfoam. The study revealed that without using gelfoam and creating a good middle ear air pocket the results were comparable to the method of using middle ear gelfoam bed. Also few insights and understanding regarding the parameters we need to asses preoperatively were also observed. With comparable healing and hearing results without using gelfoam in the middle ear, the normal physiology is restored in the postoperative period much earlier. The result of the surgery also becomes evident in the second postoperative week when the use of gelfoam in the middle ear is avoided.


To loop or not to loop? CNS-28 is the answer.
Regulation of interferon-γ is critical to constrain inflammation and mount effective responses to infection and anti-tumor immunity. In this issue of Immunity, Cui et al. identify a distal silencer element that disrupts promoter-enhancer looping, regulating IFN-γ expression and preventing inappropriate inflammation.


A multi-functional zwitterionic hydrogel with unique micro-structure, high elasticity and low modulus.
Owing to their tissue-like softness and low modulus, hydrogels minimize the mechanical mismatch with biological tissues and have received wide attention as biomaterials. However, the development of soft hydrogels is often limited by their brittleness. Here, an ultra-soft and tough hydrogel based on zwitterionic poly(sulfobetaine methacrylate) (PSBMA) was designed and successfully prepared. The obtained PSBMA hydrogel exhibits a unique spike-like micro-structure, low modulus, good stretchability and excellent compressive elasticity, due to the formation of a dual-crosslinking structure. The obtained hydrogel also possesses self-healing properties and electromechanical responses to tensile and compressive deformations. Moreover, the hydrogel has good compatibility attributed to its outstanding anti-protein-adsorption properties.


Reconstruction of Composite Soft Tissue Defect in the Distal Finger Using Partial Toenail Flap Transfer.
Composite tissue loss involving the distal finger pulp and the nail is a common but challenging finger injury to restore. This study introduces a reconstruction procedure for a distal finger pulp and nail defect using a partial toenail flap transfer. Twenty digits, including 16 thumbs, two index fingers, and two middle fingers, with composite soft tissue defects were treated with a partial toenail flap transfer from October 2015 to January 2020. Shortening revision of the great toe phalanx, a V-Y advancement flap of the toe pulp, and a local pedicle flap from a second toe transfer were used to cover the donor sites, and no skin grafts were required. Functionality was evaluated using the validated Spanish version of the Quick-DASH scale. The aesthetics of both the reconstructed and donor sites were evaluated using the Vancouver Scar Scale (VSS). The static two-point discrimination (2-PD) of the finger pulp was used as a measure of tactile agnosia. All donor site wounds healed well. The average follow-up time was 23.6 months (6-39 months). The mean Quick-DASH functional score was 7.1. The VSS scores were 4.02 ± 0.29 and 4.00 ± 0.38 for the reconstructed and donor sites, respectively. The static 2-PD of finger pulp was 4.5 ± 0.76 mm. The patients were satisfied with finger motion, sensory function, and aesthetic contour. Partial toenail flap transfer is the recommended treatment to regain motion, sensation, function, and a satisfactory aesthetic appearance when considering repairing a composite soft tissue distal finger defect with accompanying loss of the perionychium, particularly in the thumb, index finger, or middle finger.


Biosynthesis of mycophenolic acid: purification and characterization of S-adenosyl-L-methionine: demethylmycophenolic acid O-methyltransferase.
The final step in the biosynthesis of mycophenolic acid involves the transfer of a methyl group from S-adenosylmethionine to demethylmycophenolic acid. The enzyme, S-adenosylmethionine:demethylmycophenolic acid O-methyltransferase, was isolated from Penicillium stoloniferum and purified 2,700-fold by ammonium sulfate fractionation and diethylaminoethyl-cellulose and Sephadex G-200 chromatography. Maximum enzyme activity was achieved at pH 7.5 and a temperature of 27 to 28 C. The apparent K(m) for demethylmycophenolic acid was 3.1 x 10(-6) M. The enzyme preparation was 50% inactivated when exposed to 33 C for 15 min. Mycophenolic acid, homocystine, S-adenosyl-homocysteine, ethanol, and Mg(2+) inhibited the methyltransferase. This enzyme appears to be subject to end product inhibition which may regulate the synthesis of mycophenolic acid. The methyltransferase activity was highest during the early phases of the fermentation.


ROS-dependent caspase-9 activation in hypoxic cell death.
Mitochondria are known to play a fundamental role in apoptosis by releasing apoptogenic molecules such as cytochrome c into the cytoplasm, thereby sequentially activating initiator caspase-9. However, the mechanisms of cytochrome c release or caspase-9 activation in response to hypoxia are unclear. In this report, we show that caspase-9 is activated by reactive oxygen species (ROS) without involvement of cytochrome c release in hypoxic injury. In addition, activated caspase-9 induces permeability transition (PT)-independent cytochrome c release, suggesting that caspase-9 may disrupt mitochondrial diffusion limit of cytochrome c and serve to amplify further release of cytochrome c.


Morphine and self-stimulation: evidence for action on a common neural substrate.
Recent studies have demonstrated that the self-stimulation phenomenon may provide a useful technique for investigating the rewarding properties of potentially addictive drugs such as morphine. The present study attempted to examine the nature of morphine's effects on self-stimulation by observing changes in rate-intensity functions following morphine administration. The results indicate that morphine markedly enhanced bar pressing for low intensity stimulation when the intensities were presented in an ascending sequence but morphine produced only slight changes in self-stimulation rates when a descending series was used. The failure of morphine to facilitate responding in the descending series suggests that adaptation of the self-stimulation system can block morphine's effects on this system. These findings appear to support the hypothesis that morphine affects the excitability of the neural system which mediates self-stimulation.


Fortune favours the brave: Movement responses shape demographic dynamics in strongly competing populations.
Animal movement is a key mechanism for shaping population dynamics. The effect of interactions between competing animals on a population's survival has been studied for many decades. However, interactions also affect an animal's subsequent movement decisions. Despite this, the indirect effect of these decisions on animal survival is much less well-understood. Here, we incorporate movement responses to foreign animals into a model of two competing populations, where inter-specific competition is greater than intra-specific competition. When movement is diffusive, the travelling wave moves from the stronger population to the weaker. However, by incorporating behaviourally induced directed movement towards the stronger population, the weaker one can slow the travelling wave down, even reversing its direction. Hence movement responses can switch the predictions of traditional mechanistic models. Furthermore, when environmental heterogeneity is combined with aggressive movement strategies, it is possible for spatially segregated co-existence to emerge. In this situation, the spatial patterns of the competing populations have the unusual feature that they are slightly out-of-phase with the environmental patterns. Finally, incorporating dynamic movement responses can also enable stable co-existence in a homogeneous environment, giving a new mechanism for spatially segregated co-existence.


Determination of picogram levels of brain serotonin by a simplified liquid chromatographic electrochemical detection assay.
A simplified assay is described for measurement of picogram amounts of serotonin (5-HT) by high pressure liquid chromatography with electrochemical detection (LCEC). The procedure involves pre-purification of brain 5-HT by adsorption on Amberlite CG-50. Serotonin is subsequently resolved and detected by LCEC on Zipax SCX resin. The present method gives working sensitivities of at least 100 pg, tissue recoveries of 95% and very low interassay variability (coefficient of variation = 3%). Determination of rat brain area 5-HT by this LCEC method is described and compared to other high sensitivity methods.


[Psoriasis and bullous pemphigoid. Description of 3 clinical cases].
Three cases with coexisting psoriasis and bullous pemphigoid are reported and the possible etiopathogenetic relationships between the two diseases are discussed.


Influence of low levels of water salinity on toxicity of nitrite to anuran larvae.
Reactive nitrogen compounds such as nitrite (NO2(-)) are highly toxic to aquatic animals and are partly responsible for the global decline of amphibians. On some fish and Caudata amphibian species low levels of sodium chloride significantly reduce the toxicity of nitrite. However, the nitrite-salinity interaction has not been properly studied in anuran amphibians. To verify if chloride (Cl(-)) attenuates NO2(-) toxicity, eggs and larvae of three anuran species were subjected to a series of NO2(-) solutions combined with three salt concentrations (0, 0.4 and 2 or 0, 0.052 and 0.2gL(-1)NaCl). One of the species tested originated from two different populations inhabiting highly contrasted nutrient richness environments: lowland Doñana Natural Park and Sierra de Gredos Mountain. In general, the presence of Cl(-) increased survival and growth of lowland Pelophylax perezi and activity of mountain P. perezi larvae exposed to NO2(-), thus attenuating the toxicity of NO2(-) to developing amphibians. Mountain amphibian populations appeared to be much more sensitive to the concentrations of NO2(-) and Cl(-) used in this experiment than coastal conspecifics, suggesting possible adaptation of populations to local conditions. Nitrogen pollution in coastal wetlands poses a serious threat to aquatic organisms, causing direct toxicity or indirect effects via ecosystem eutrophication. The presence of low to medium levels of salinity that would be common in coastal wetlands may attenuate the direct effects of increasing concentrations of nitrogenous compounds in water bodies. Furthermore, treating cultures of endangered anurans with small amounts of NaCl may provide an additional protective measure.


Whole-Body Hypothermia, Cerebral Magnetic Resonance Biomarkers, and Outcomes in Neonates With Moderate or Severe Hypoxic-Ischemic Encephalopathy Born at Tertiary Care Centers vs Other Facilities: A Nested Study Within a Randomized Clinical Trial.
The association between place of birth and hypothermic neuroprotection after hypoxic-ischemic encephalopathy (HIE) in low- and middle-income countries (LMICs) is unknown. To ascertain the association between place of birth and the efficacy of whole-body hypothermia for protection against brain injury measured by magnetic resonance (MR) biomarkers among neonates born at a tertiary care center (inborn) or other facilities (outborn). This nested cohort study within a randomized clinical trial involved neonates at 7 tertiary neonatal intensive care units in India, Sri Lanka, and Bangladesh between August 15, 2015, and February 15, 2019. A total of 408 neonates born at or after 36 weeks' gestation with moderate or severe HIE were randomized to receive whole-body hypothermia (reduction of rectal temperatures to between 33.0 °C and 34.0 °C; hypothermia group) for 72 hours or no whole-body hypothermia (rectal temperatures maintained between 36.0 °C and 37.0 °C; control group) within 6 hours of birth, with follow-up until September 27, 2020. 3T MR imaging, MR spectroscopy, and diffusion tensor imaging. Thalamic N-acetyl aspartate (NAA) mmol/kg wet weight, thalamic lactate to NAA peak area ratios, brain injury scores, and white matter fractional anisotropy at 1 to 2 weeks and death or moderate or severe disability at 18 to 22 months. Among 408 neonates, the mean (SD) gestational age was 38.7 (1.3) weeks; 267 (65.4%) were male. A total of 123 neonates were inborn and 285 were outborn. Inborn neonates were smaller (mean [SD], 2.8 [0.5] kg vs 2.9 [0.4] kg; P = .02), more likely to have instrumental or cesarean deliveries (43.1% vs 24.7%; P = .01), and more likely to be intubated at birth (78.9% vs 29.1%; P = .001) than outborn neonates, although the rate of severe HIE was not different (23.6% vs 17.9%; P = .22). Magnetic resonance data from 267 neonates (80 inborn and 187 outborn) were analyzed. In the hypothermia vs control groups, the mean (SD) thalamic NAA levels were 8.04 (1.98) vs 8.31 (1.13) among inborn neonates (odds ratio [OR], -0.28; 95% CI, -1.62 to 1.07; P = .68) and 8.03 (1.89) vs 7.99 (1.72) among outborn neonates (OR, 0.05; 95% CI, -0.62 to 0.71; P = .89); the median (IQR) thalamic lactate to NAA peak area ratios were 0.13 (0.10-0.20) vs 0.12 (0.09-0.18) among inborn neonates (OR, 1.02; 95% CI, 0.96-1.08; P = .59) and 0.14 (0.11-0.20) vs 0.14 (0.10-0.17) among outborn neonates (OR, 1.03; 95% CI, 0.98-1.09; P = .18). There was no difference in brain injury scores or white matter fractional anisotropy between the hypothermia and control groups among inborn or outborn neonates. Whole-body hypothermia was not associated with reductions in death or disability, either among 123 inborn neonates (hypothermia vs control group: 34 neonates [58.6%] vs 34 [56.7%]; risk ratio, 1.03; 95% CI, 0.76-1.41), or 285 outborn neonates (hypothermia vs control group: 64 neonates [46.7%] vs 60 [43.2%]; risk ratio, 1.08; 95% CI, 0.83-1.41). In this nested cohort study, whole-body hypothermia was not associated with reductions in brain injury after HIE among neonates in South Asia, irrespective of place of birth. These findings do not support the use of whole-body hypothermia for HIE among neonates in LMICs. ClinicalTrials.gov Identifier: NCT02387385.


Local delivery of hyaluronic acid as an adjunct to scaling and root planing in the treatment of chronic periodontitis in smokers and non-smokers: A clinical and microbiological study.
The objective of the study is to assess the effects of adjunctive use of hyaluronic acid (HA), clinically and microbiologically in smokers and nonsmokers, in the management of chronic periodontitis. 48 sites from 24 chronic periodontitis patients, including smokers and nonsmokers with probing depth >5 mm, were selected for the study. A split-mouth design was followed. Scaling and root planing (SRP) was performed for all the patients. The test sites received 0.8% HA gel whereas the control site received placebo gel. The clinical parameters plaque, gingival and bleeding indices, Pocket Probing Depth (PPD), and clinical attachment level (CAL) were evaluated at baseline, 1 and 3 months. Microbiological parameters were evaluated at baseline and 1 month. The results showed reduction in PPD and gain in CAL in both smokers and nonsmokers at the end of 3rd month. The improvements in the test sites were statistically significant when compared with that of control sites. The microbiological analysis showed a significant reduction in Aggregatibacter actinomycetemcomitans and Porphyromonas gingivalis at the test sites when compared to the controls in both the groups. The findings in both groups suggest that the use of locally delivered 0.8% HA gel may be valuable as an adjunct to SRP in the treatment of chronic periodontitis.


A novel Hg2+ selective ratiometric fluorescent chemodosimeter based on an intramolecular FRET mechanism.
A irreversible Hg2+ selective ratiometric fluorescence probe FR, a fluorescein fluorophore linked to a rhodamine B hydrazide by a thiourea spacer, was designed and synthesized. The developed probe FR exhibited great ratiometric fluorescence enhancement and remarkable yellow-magenta color change toward Hg2+ with excellent selectivity in aqueous acetone solution, and the ratiometric fluorescence response to Hg2+ was not interfered by other metal cations including Fe3+, Co2+, Ni2+, Cr3+, Zn2+, Pb2+, Cd2+, Ca2+, Mg2+, Ba2+ and Mn2+. The linear range and the detection limit of this supposed ratiometric fluorescence method for Hg2+ were 0.0-10.0x10(-6) and 5x10(-8) M, respectively.


Somatostatin Neurons Govern Theta Oscillations Induced by Salient Visual Signals.
Salient visual stimuli enhance theta oscillations and spike-phase locking in the theta band in the primary visual cortex (V1) of mice; however, the detailed mechanisms remain unknown. GABAergic neurons play a vital role in regulating these oscillations. Here, we use optogenetic recordings to tag cell-type-specific neurons in V1 of head-fixed mice and demonstrate that salient visual stimuli facilitate somatostatin (SOM)-expressing neuron responses and firing with theta band oscillations but suppress activities of parvalbumin (PV)-expressing neurons. Furthermore, inactivation of SOM neurons attenuates the enhancement of theta oscillations induced by salient visual stimuli and rhythmic activation of SOM neurons enhances theta oscillations. These results reveal a potential cortical theta oscillation mechanism governed by SOM neurons.


EGF, GDNF, and IGF-1 influence the proliferation and stemness of ovine spermatogonial stem cells in vitro.
The objective of the present study was to purify sheep spermatogonial stem cells (SSCs) from testicular isolate using combined enrichment methods and to study the effect of growth factors on SSC stemness during culture. The testicular cells from prepubertal male sheep were isolated, and SSCs were purified using Ficoll gradients (10 and 12%) followed by differential plating (laminin with BSA). SSCs were cultured with StemPro®-34 SFM, additives, and FBS for 7 days. The various doses (ng/ml) of growth factors, EGF at 10, 15, and 20, GDNF at 40, 70, and 100 and IGF-1 at 50, 100, and 150 were tested for the proliferation and stemness of SSCs in vitro. The stemness in cultured cells was assessed using SSC markers PLZF, ITGA6, and GFRα1. Ficoll density gradient separation significantly (p < 0.05) increased the percentage of SSCs in 12% fraction (35.1 ± 3.8 vs 11.2 ± 3.7). Subsequently, purification using laminin with BSA plating further enriched SSCs to 61.7 ± 4.7%. GDNF at 40 ng/ml, EGF at 15 and 20 ng/ml and IGF1 at 100 and 150 ng/ml significantly (p < 0.05) improved proliferation and stemness of SSCs up to 7 days in culture. GDNF at 40 ng/ml outperformed other growth factors tested and could maintain the ovine SSCs proliferation and stemness for 36 days. The combined enrichment method employing density gradient centrifugation and laminin with BSA plating improves the purification efficiency of ovine SSCs. GDNF at 40 ng/ml is essential for optimal proliferation and sustenance of stemness of ovine SSCs in vitro.


alpha1,3 fucosyltransferase-VII up-regulates the mRNA of alpha5 integrin and its biological function.
After transfection of alpha1,3fucosyltransferase (FucT)-VII cDNA into H7721 human hepatocarcinoma cells, the expression of alpha5, but not beta1 integrin was significantly up-regulated. This was evidenced by the increase of alpha5 integrin on cell surface as well as the increase of alpha5 mRNA and protein in the cells. However, the expressions of sialyl Lewis X (SLe(x), the product of alpha1,3FucT-VII) on both alpha5 and beta1 integrin subunits were unchanged. Concomitantly, the tyrosine autophosphorylated FAK and dephosphorylated Src (FAK and Src involve in the signal transduction of integrin alpha5beta1) were up-regulated, while the Tyr-527 phosphorylated Src was down-regulated. The above-mentioned alterations were correlated to the expressions of alpha1,3FucT-VII in different alpha1,3FucT-VII transfected H7721 cell lines. In addition, after alpha1,3FucT-VII transfection, cell adhesion to fibronectin (Fn) and chemotaxic cell migration were obviously promoted. The cell adhesion could be blocked by alpha5 integrin antibody, and cell migration was obviously attenuated by the antibodies to both alpha5 integrin and SLe(x). These findings suggest that the increased surface alpha5 integrin caused by the up-regulation of alpha5 mRNA promotes the cell adhesion to Fn, cell migratiom, and Fn-induced signaling of alpha5beta1 integrin. The up-regulation of surface SLe(x) originated from the over expression of alpha1,3FucT-VII also led to the stimulation of cell migration. This is the first time to report that alpha1,3FucT-VII can regulate the mRNA expression of integrin.


Nprl3 is required for normal development of the cardiovascular system.
C16orf35 is a conserved and widely expressed gene lying adjacent to the human α-globin cluster in all vertebrate species. In-depth sequence analysis shows that C16orf35 (now called NPRL3) is an orthologue of the yeast gene Npr3 (nitrogen permease regulator 3) and, furthermore, is a paralogue of its protein partner Npr2. The yeast Npr2/3 dimeric protein complex senses amino acid starvation and appropriately adjusts cell metabolism via the TOR pathway. Here we have analysed a mouse model in which expression of Nprl3 has been abolished using homologous recombination. The predominant effect on RNA expression appears to involve genes that regulate protein synthesis and cell cycle, consistent with perturbation of the mTOR pathway. Embryos homozygous for this mutation die towards the end of gestation with a range of cardiovascular defects, including outflow tract abnormalities and ventriculoseptal defects consistent with previous observations, showing that perturbation of the mTOR pathway may affect development of the myocardium. NPRL3 is a candidate gene for harbouring mutations in individuals with developmental abnormalities of the cardiovascular system.


Isocratic reversed-phase high-performance liquid chromatography of ribonucleotides, deoxynucleotides, cyclic nucleotides and deoxycyclic nucleotides.
An isocratic reversed-phase high-performance liquid chromatographic system is described for the separation of a relatively large number (greater than 20) of ribonucleotides, deoxynucleotides, cyclic nucleotides and deoxycyclic nucleotides. A 25 cm x 5 mm ODS-Hypersil (5 microns particle size) column was used with methanol-triethylammonium phosphate buffer as eluent. The effects of methanol content, pH, and ionic strength of the buffer on retention and resolution of the nucleotides have been studied. The applicability of the system was demonstrated by the analysis of nucleotides in cells and tissue extracts.


Bronchodilating activity of oral clenbuterol in asthmatic children after single administration of different dosages.
The dose-response activity of clenbuterol, a new long-acting beta 2-agonist, was evaluated in 12 children aged 5 to 11 years with moderate to severe asthma. The study was a double-blind cross-over comparison of three oral dose levels: 0.5, 1.0, and 1.5 micrograms/kg and placebo all in the form of syrup. Pulmonary function, heart rate, blood pressure, and tremor were evaluated at 30, 60, 90, and 120 min, then hourly for 8 hr, following the ingestion of the drug. The bronchodilating effect of clenbuterol, evaluated as percentage changes in expiratory flow rates was significantly different from placebo. The overall fall-off in the effect of 1.0 micrograms/kg and 1.5 micrograms/kg doses after 8 hr was small. The results of the 1.0 micrograms/kg dose often overlapped those of the 1.5 micrograms/kg dose, suggesting that a single 1.0 micrograms/kg dose of clenbuterol is most advisable in children, assuring the most favorable risk/benefit ratio. Even the highest dose of clenbuterol caused only a marginal increase in tremor, not statistically different from that induced by placebo.


Are personalities genetically determined? Inferences from subsocial spiders.
Recent research has revealed that polymorphic behavioral strategies shape intra-and interspecific interactions and contribute to fitness in many animal species. A better understanding of the proximate mechanisms underlying these behavioral syndromes will enhance our grasp this phenomenon. Spiders in the genus Anelosimus exhibit inter-individual behavioral variation on several axes: individuals have consistent responses to stimuli (e.g. bold vs. shy individuals) and they are subsocial (exhibiting extended maternal care and sibling cooperation) across most of their range, but they sometimes form permanent social groups in northern temperate regions. Here, we seek genetic variants associated with boldness and with social structure in a socially polymorphic population of the spider Anelosimus studiosus. We also develop preliminary genomic resources, including a genome assembly and linkage map, that support this and future genomic research on this group. Remarkably, we identify a small genomic scaffold (~ 1200 bp) that harbors seven single nucleotide polymorphisms (SNPs) associated with boldness. Moreover, heterozygotes are less common than expected based on Hardy-Weinberg equilibrium, suggesting that either assortative mating or selection against heterozygotes may be occurring in this system. We find no loci significantly associated with social organization. Our draft genome assembly allows us to localize SNPs of interest in this study and to carry out genetic comparisons with other published genomes, although it remains highly fragmented. By identifying a locus associated with a well-studied animal personality trait, this study opens up avenues for future research to link behavioral studies of animal personality with genotype and fitness.


Peracute and acute death from ischaemic heart disease within 4 days of onset of symptoms.
Post-mortem examination was made of 136 patients who died of ischaemic heart disease from within a few minutes to 4 days after the onset to symptoms. Acute myocardial infarction was detected by means of the histochemistry of dehydrogenases in 102 cases (75%); in 34 cases(25%) no infarction was found, but the presence of coronary obstruction together with clinical symptoms justified the diagnosis of acute coronary insufficiency. Fifty-six of the infarctions occurred within 2--3 h after the onset of ischaemia, 24 within 4--5 h, 12 within 6--24 h, and 10 within 1--4 days. The chances of recovery increased if the patients survived for 24 h. The period of 5 h after the onset of ischaemia appeared especially important; 80 of 102 patients with acute myocardial infarction died during that period. Thirty-one of 34 patients with acute coronary insufficiency and 60 of 102 patients with acute myocardial infarction died peracutely, i.e. within a few minutes to 2 h after the onset of symptoms. In the patients of the latter group the onset of symptoms must have been delayed, otherwise none of these patients could have appeared in the category of peracute death. This delay was more obvious in the patients with acute myocardial infarction localized in the area supplied by the left descending artery. The occurrence of peracute deaths could not be correlated witha particular localization of area of infarction. The causes of coronary obstructions and the role of dynamic disturbances of coronary perfusion in the pathogenesis of coronary insufficiency and myocardial infarction and discussed.


Computational workflow for integrative analyses of DNA replication timing, epigenomic, and transcriptomic data.
Temporal profiling of DNA replication timing (RT) in combination with chromatin modifications, chromatin accessibility, and gene expression provides new insights into the causal relationships between chromatin and RT during cell cycle. Here, we describe a protocol for in-depth integrative computational analyses of Repli-seq, ATAC-seq, RNA-seq, and ChIP-seq or CUT&RUN data for multiple marks at various time points across cell cycle and changes in their interrelationships upon an experimental perturbation (e.g., knockdown or overexpression of a regulatory protein). For complete details on the use and execution of this protocol, please refer to Van Rechem et al. (2021).


Elevated plasma glutamate levels in colorectal carcinoma patients and in patients with acquired immunodeficiency syndrome (AIDS).
Amino acid concentrations were analyzed in the sera of HIV (LAV/HTLV-III) positive persons and in the plasma of colorectal carcinoma patients. Both groups of persons showed significantly elevated glutamate concentrations when compared with healthy control persons. Glutamate concentrations were found to be strongly elevated in all groups of HIV-positive persons including patients with acquired immunodeficiency syndrome (AIDS) or lymphadenopathy as well as HIV-positive persons without overt symptoms, indicating that increased plasma glutamate levels may be among the earliest consequences of the HIV infection. Moreover, the increased plasma glutamate concentrations in the colorectal carcinoma patients were correlated with a decreased immunological reactivity (mitogenic responses against concanavalin A). This suggests the possibility that the increased plasma glutamate concentrations may be causally responsible for the decreased immunological reactivity in colorectal carcinoma patients as well as in patients with AIDS.


Botulinum neurotoxin type A for the treatment of pain: not just in migraine and trigeminal neuralgia.
Despite their huge epidemiological impact, primary headaches, trigeminal neuralgia and other chronic pain conditions still receive suboptimal medical approach, even in developed countries. The limited efficacy of current pain-killers and prophylactic treatments stands among the main reasons for this phenomenon. Botulinum neurotoxin (BoNT) represents a well-established and licensed treatment for chronic migraine, but also an emerging treatment for other types of primary headache, trigeminal neuralgia, neuropathic pain, and an increasing number of pain conditions. We searched and critically reviewed evidence for the efficacy of BoNT for the treatment of chronic pain. Meta-analyses and randomized controlled trials (RCTs) suggest that BoNT potentially represents a multi-purpose drug for the treatment of pain in several disorders due to a favorable safety profile and a long-lasting relief after a single injection. BoNT is an emerging treatment in different pain conditions. Future RCTs should explore the use of BoNT injection therapy combined with systemic drugs and/or physical therapies as new pain treatment strategies.


MicroRNA regulation and analytical methods in cancer cell metabolism.
The reprogramming of glucose metabolism from oxidative to glycolytic metabolism, known as the Warburg effect, is an anomalous characteristic of cancer cell metabolism. Recent studies have revealed a subset of microRNAs (miRNAs) that play critical roles in regulating the reprogramming of glucose metabolism in cancer cells. These miRNAs regulate cellular glucose metabolism by directly targeting multiple metabolic genes, including those encoding key glycolytic enzymes. In the first part of this review, we summarized the recent knowledge of miRNA regulation in the reprogramming of glucose metabolism in cancer cells and discussed the potential utilization of the key miRNA regulators as metabolic targets for developing new antitumor agents. Then, we summarized recent advances in methods and techniques for studying miRNA regulation in cancer cell metabolism.


DNA sequences bound specifically by glucocorticoid receptor in vitro render a heterologous promoter hormone responsive in vivo.
Glucocorticoids stimulate transcriptional initiation within integrated mammary tumor virus (MTV) DNA sequences in infected cells. We report here that production of herpes simplex virus thymidine kinase (tk) RNA is stimulated as much as 50-fold by the glucocorticoid, dexamethasone, when sequences from a particular region of MTV DNA are fused upstream of the normally constitutive tk promoter region. Three cloned fragments of MTV DNA were tested, each itself lacking the sequences required for transcription initiation. We monitored the effects of dexamethasone on the efficiency with which these recombinants transfect a tk- rat cell line to a tk+ phenotype, and measured tk enzymatic activity, the size and abundance of tk mRNA, and the 5' termini of tk transcripts in the transfectants. We conclude that the MTV promoter region contains a "glucocorticoid response element" that can be separated from a second element essential for MTV transcription initiation. The hormone response element maps within a 340-base pair MTV DNA fragment that contains specific binding sites for purified glucocorticoid receptor protein in vitro, implying that receptor binding at these sites in vivo may mediate hormone responsiveness. Comparison of several different constructions indicates that the location and orientation of the glucocorticoid response element relative to the transcription start site is not rigidly constrained.


Comparison of the rebound tonometer (TonoVet) with the applanation tonometer (TonoPen XL) in normal Eurasian Eagle owls (Bubo bubo).
To examine the feasibility and accuracy of a handheld rebound tonometer, TonoVet, and to compare the intraocular pressure (IOP) readings of the TonoVet with those of an applanation tonometer, TonoPen XL, in normal Eurasian Eagle owls. Ten clinically normal Eurasian Eagle owls (20 eyes). Complete ocular examinations, using slit-lamp biomicroscopy and indirect ophthalmoscopy, were conducted on each raptor. The IOP was measured bilaterally using a rebound tonometer followed by a topical anesthetic agent after 1 min. The TonoPen XL tonometer was applied in both eyes 30 s following topical anesthesia. The mean +/- SD IOP obtained by rebound tonometer was 10.45 +/- 1.64 mmHg (range 7-14 mmHg), and by applanation tonometer was 9.35 +/- 1.81 mmHg (range 6-12 mmHg). There was a significant difference (P = 0.001) in the IOP obtained from both tonometers. The linear regression equation describing the relationship between both devices was y = 0.669x + 4.194 (x = TonoPen XL and y = TonoVet). The determination coefficient (r(2)) was r(2) = 0.550. The results suggest that readings from the rebound tonometer significantly overestimated those from the applanation tonometer and that the rebound tonometer was tolerated well because of the rapid and minimal stress-inducing method of tonometry in the Eurasian Eagle owls, even without topical anesthesia. Further studies comparing TonoVet with manometric measurements may be necessary to employ rebound tonometer for routine clinical use in Eurasian Eagle owls.


Immunomodulatory mechanisms during Echinococcus granulosus infection.
The pathologic events that ensue after humans ingest the eggs of Echinococcus granulosus and continue while cystic echinococcosis develops, provide an excellent example illustrating the evasive strategies helminth parasites use to develop, progress and cause chronic disease. The hydatid cyst secretes and exposes numerous immunomodulatory molecules to the host's immune system. By characterizing these molecules we can understand the mechanisms that E. granulosus uses for increasing the efficiency and persistency of infection in the host. These molecules modulate both the innate and adaptive arms of the immune response and appear to target cellular and humoral responses. In this review, we discuss recent advances in the immunobiology of host-E. granulosus interactions that provide intriguing insights into the complex interplay between host and parasite that ultimately facilitates parasite survival.


Does exposure to testosterone significantly alter endogenous metabolism in the marine mussel Mytilus galloprovincialis?
Mussels (Mytilus galloprovincialis) were exposed to different concentrations of testosterone (T: 20, 200 and 2000ng/L) in a semi-static water regime (1-day dosing intervals) for up to 5 days in an attempt to see whether endogenous steroid levels and steroid metabolism were altered by exogenous exposure to testosterone. Whole tissue levels of total testosterone (free+esterified) sharply increased in a concentration-dependent manner, from 2ng/g in controls to 290ng/g in organisms exposed to the highest concentration. In contrast, levels of free testosterone were only significantly elevated at the high-exposure group (5-fold increase with respect to controls). Increased activity of palmitoyl-CoA:testosterone acyltransferase (ATAT) was detected in organisms exposed to the highest concentration of testosterone, while those exposed to low and medium concentrations showed significant alterations in their polyunsaturated fatty acid profiles. The obtained results suggest that esterification of the excess of T with fatty acids might act as a homeostatic mechanism to maintain endogenous levels of free T stable. Interestingly, a decrease in CYP3A-like activity was detected in T-exposed mussels together with a significant decrease in the metabolism of the androgen precursor androstenedione to dihydrotestosterone (5α-DHT). Overall, the work contributes to the better knowledge of androgen metabolism in mussels.


An immunohistochemical study of gelsolin immunoreactivity in corneal amyloidosis.
A variant of the actin-modulating protein gelsolin has recently been identified as a component of the amyloid deposits in familial amyloidosis, Finnish type (Meretoja's syndrome), and has been demonstrated immunohistochemically in amyloid deposits in the cornea, and in the skin, kidney, heart, thyroid gland, salivary gland, and rectum of patients with this disease. With the use of immunohistochemistry involving an antibody against gelsolin, we examined a corneal specimen from a patient with Meretoja's syndrome and 14 corneal specimens with lattice dystrophy type I, atypical lattice dystrophy, polymorphic amyloid degeneration, primary familial amyloidosis, or secondary corneal amyloidosis. Our results showed the presence of a gelsolin-related protein either within or around corneal amyloid deposits in nine of the 15 specimens and markedly increased anti-gelsolin immunoreactivity of the corneal keratocytes in 13 of the 15 diseased corneas. These data indicated that the accumulation of gelsolin may be seen in various forms of amyloidosis and may not be confined to Meretoja's syndrome.


Effects of pharmacological inhibition of hyaluronic acid synthesis on experimental endometriosis.
Dysregulated hyaluronic acid (HA) metabolism has been shown to be implicated in several pathologies including endometriosis. 4-Methylumbelliferone (4MU) is an HA synthesis inhibitor with proven antitumour activity. In this study, we aim to evaluate the effect of 4MU on endometriosis development both in vivo and in vitro. Endometriosis was surgically induced by uterine tissue auto-transplantation in 32 two-month-old BALB/c mice. Animals were designated into the early or late starting treatment group, which initiated on day 2 or day 15 after surgery, respectively. Within each group, 4MU 200 mg/kg/day or vehicle (Control) were administered by oesophageal gavage for 28 days. After sacrifice, the percentage of developed lesions, lesion size, cell proliferation, vascularization and HA deposition within the endometriotic-like lesions were evaluated. Cell viability was assessed in endometrial epithelial cells (ECC-1) and in endometrial stromal cells (t-HESC); and migration was evaluated in t-HESC. There was a significant reduction in the percentage of developed lesions in mice that started the 4MU treatment on day 2 compared with its respective control group, and compared with those that started treatment on day 15. However, no significant changes were found when analysing endometriotic-like lesion's cell proliferation, vascularization and HA deposition. In vitro, both cell viability and migration were inhibited by 4MU treatment. The inhibition of HA synthesis could be a beneficial and alternative option to treat endometriosis at the early stage of the disease. Further research is necessary to elucidate 4MU's mechanism of action and better strategies for delivering this promising drug.


Autoregulation of the tufB operon in Salmonella.
In Salmonella enterica and related species, translation elongation factor EF-Tu is encoded by two widely separated but near-identical genes, tufA and tufB. Two thirds of EF-Tu is expressed from tufA with the remaining one third coming from tufB. Inactivation of tufA is partly compensated by a doubling in the amount of EF-TuB but the mechanism of this up-regulation is unknown. By experimental evolution selecting for improved growth rate in a strain with an inactive tufA we selected six different noncoding or synonymous point mutations close to the tufB start codon. Based on these results we constructed a total of 161 different point mutations around the tufB start codon, as well as tufB 3'-truncations, and measured tufB expression using tufB-yfp transcriptional and translational fusions. The expression data support the presence of two competing stem-loop structures that can form in the 5'-end of the tufB mRNA. Formation of the 'closed' structure leads to Rho-dependent transcriptional termination of the tufB mRNA. We propose a model in which translational speed is used as a sensor for EF-Tu concentration and where the expression of tufB is post-transcriptionally regulated. This model describes for the first time how expression of the most abundant Salmonella protein is autoregulated.


MDM2-positive papillary sarcomatoid renal cell carcinoma: a potential diagnostic pitfall.
Sarcomatoid renal cell carcinoma is a highly aggressive form of carcinoma, histologically showing both carcinomatous and mesenchymal component in different proportions. We present a case of advanced type 1 papillary sarcomatoid renal cell carcinoma infiltrating adjacent organs and showing positivity for MDM2 by immunohistochemistry and MDM2 amplification by fluorescence in situ hybridization. This finding, together with sarcomatoid morphology, poses a potential pitfall for diagnosis with dedifferentiated liposarcoma. MDM2 is known to be altered in various human sarcomas. Only recently, MDM2 alterations have been reported in carcinomas. The presented case illustrates the need of thorough sampling with clinic-pathological correlation before making a final diagnosis in sarcomatoid retroperitoneal tumours. Additionally, the potential clinical implications of MDM2 amplification in renal cell carcinoma are discussed.


Gene therapy: some history, applications, problems, and prospects.
The concept of transferring genes to tissues for clinical applications has been discussed for nearly half a century, but our ability to manipulate genetic material via recombinant DNA technology has brought this goal to reality. While originally conceived as a way to treat life-threatening disorders (inborn errors, cancers) refractory to conventional treatment, gene therapy now is considered for many non-life-threatening conditions, including those adversely affecting a patient's quality of life. The lack of suitable treatment has become a rational basis for extending the scope of gene therapy. This manuscript reviews the general methods by which genes are transferred as well as diverse examples of clinical applications (acquired tissue damage, upper gastrointestinal tract infection, autoimmune disease, systemic protein deficiency). Despite some well-publicized problems, gene therapy has made substantive progress, including tangible success, albeit much slower than was initially predicted. Although gene therapy is still at a fairly primitive stage, it is firmly science based. There is justifiable optimism that with increased pathobiological understanding and biotechnological improvements, gene therapy will become a standard part of clinical practice within 20 years.


Dispersion-Controlled Regioselective Acid-Catalyzed Intramolecular Hydroindolation of cis-Methindolylstyrenes To Access Tetrahydrobenzo[ cd]indoles.
Readily prepared cis-β-(α',α'-dimethyl)-4'-methindolylstyrenes undergo acid-catalyzed intramolecular hydroindolation to afford tetrahydrobenzo[ cd]indoles. Our experimental and computational investigations suggest that dispersive interactions between the indole and styrene preorganize substrates such that 6-membered ring formation is preferred, apparently via concerted protonation and C-C bond formation. When dispersion is attenuated (by a substituent or heteroatom), regioselectivity erodes and competing oligomerization predominates for cis substrates. Similarly, all trans-configured substrates that we evaluated failed to cyclize efficiently.


Multidimensional analysis of the learning curve for laparoscopic resection in rectal cancer.
We attempted to assess the learning curve for laparoscopic resection for rectal cancer. We included 381 patients who underwent laparoscopic resection for rectal cancer between December 2002 and December 2007. The operative experience was divided into four periods according to numbers of operations and significant changes in main surgical results. Operative time decreased significantly after 90 operations. The overall anastomotic leakage rate was 3.7%; 14.6% for the first 50 patients and 5.4% for the following 40 patients. The overall conversion rate was 2.9%, 4-6% during the first and second periods, but decreasing thereafter. The number of harvested lymph nodes and distal resection margin was within an acceptable range during the entire period. For the patients with stage I-III tumors, the local recurrence rate was 4.4% and the overall recurrence rate was 22.9%. The local recurrence rate was 8.9% initially and decreasing to 1.4% after the second period. The cumulative incidence of local recurrence decreased to less than 7% after 120 patients and to less than 5% after 180 cases. The learning curve for laparoscopic surgery for rectal cancer changed over time. Moreover, the learning curve for oncological safety was longer than that for operative safety.


Expression of NMU, PPBP and GNG4 in colon cancer and their influences on prognosis.
This study aims to identify the core genes that influence the prognosis of colon cancer (CC) and analyze their relationships with clinical characteristics. The gene expression profiles were downloaded from The Cancer Genome Atlas (TCGA) database. Differentially expressed genes (DEGs) were identified. The top ten core genes were selected by bioinformatics tools and screened through the Oncomine database. The expression of core genes in CC tissues and cells was validated by immunohistochemistry, immunoblotting and quantitative real-time polymerase chain reaction. Spearman correlation was used to analyze the relationship between different parameters. Overall survival was assessed by the Kaplan-Meier method. The area under the curve (AUC) and the receiver operating curve (ROC) were applied to assess the accuracy of genes for predicting prognosis. There were 1,665 DEGs that were identified from TCGA database. Bioinformatics analysis found that GNGT1, NMU, PPBP, AGT, and GNG4 were differentially expressed in CC tissue. Overexpression of NMU, PPBP, AGT, and GNG4 in CC was associated with shortened survival time (P<0.05). In the validation studies, the high expression levels of NMU, PPBP and GNG4 in CC cells and tissues were confirmed compared to the control groups (P<0.05) and were adverse prognostic biomarkers (P<0.01). The combination prognostic model of the three core genes predicted the 1-, 3-, and 5-year survival of CC with AUCs of 0.868, 0.635 and 0.770, respectively. High levels of NMU, PPBP, and GNG4 were associated with poor prognosis in CC. The combination prognostic model of these three genes could be a new option.


Prenatal diagnosis of penoscrotal transposition with 2- and 3-dimensional ultrasonography.
The purpose of this study was to determine the prenatal diagnostic accuracy of 2-dimensional ultrasonography (2DUS) alone versus 2DUS in combination with 3-dimensional ultrasonography (3DUS) for penoscrotal transposition. Fetuses suspected of having penoscrotal transposition on the basis of ultrasonographic findings or a family history were examined by 2DUS and then 3DUS. Of a total of 22 fetuses, 14 had penoscrotal transposition at birth. The combination of 2DUS and 3DUS correctly identified more of the penoscrotal transposition cases than 2DUS alone (90.9% versus 63.6%; P < .05). The combination of 2DUS and 3DUS improved the prenatal detection rate for penoscrotal transposition compared with 2DUS alone.


Cutaneous squamous cell carcinoma and mortality in end-stage renal disease.
In the general population, cutaneous squamous cell carcinoma (cSCC) is associated with increased all-cause mortality. Transplant patients have been shown to have an increased risk of developing cSCC, and their cSCC is associated with an increased risk for mortality. In end-stage renal disease (ESRD) patients, there is extensive mortality and immune dysfunction. Because of this immune system dysfunction, we examined whether cSCC is associated with increased risk of all-cause mortality among ESRD patients, as well as the risk factors for cSCC. We analyzed ESRD patients in the United States Renal Data System from 2004-2014, excluding organ transplant recipients. We assessed mortality using a Cox Proportional Hazards (CPH) model to control for various demographic and clinical parameters, identified using international classification of diseases (ICD)-9 codes. Of the 1,035,193 patients included, 624 (0.1%) were diagnosed with cSCC. The median survival time for those with cSCC was 3.91 years [95% confidence interval (CI) = 3.67-4.15], versus 2.92 years [95%CI = 2.92-2.93] for patients without cSCC. ESRD patients with cSCC were at lower risk of death [adjusted hazard ratio = 0.75; 95%CI = 0.69-0.82] compared to those without. Decreased risk of death was also associated with parameters such as black race, Hispanic ethnicity, tobacco dependence and actinic keratosis. Increased mortality risk was associated with increasing age, male sex, hemodialysis (versus peritoneal dialysis) and alcohol dependence. Contrary to expectations, ESRD patients with a cSCC diagnosis showed reduced all-cause mortality risk relative to those without. The reason for this discrepancy remains unclear, suggesting the need for further study.