Initial Time Point Research Articles

Biogas from Tannery Solid Waste Anaerobic Digestion Is Driven by the Association of the Bacterial Order Bacteroidales and Archaeal Family Methanosaetaceae.

The search for renewable energies has been one of the biggest challenges of the last decades. Sludge and solid wastes of many sources have been used to produce biogas of high calorific value. Thus, this work aimed to evaluate the biogas production of solid waste originating from a tannery that uses chromium salts as a tanning agent and to characterize the physicochemical parameters and microbial composition of the biogas-producing biomass. Wastes were collected and the parameters were evaluated at the initial and final time points of the anaerobic incubation process. At the end of 150days, there was a production of 26.1mLg-1 VSS of biogas with 52% of methane. The highest amount of biomethane observed was related to the archaeal family Methanosaetaceae and bacterial order Bacteroidales. Knowledge about changes in the microbial composition can provide tools for manipulation, isolation, and inoculation of the microorganisms inside the bioreactors to maximize methane production.

Complement initiation and regulation differ by sex in intestinal ischemia reperfusion injury

Abstract Males and females frequently sustain distinct symptoms during ischemic events and subsequent inflammatory events. The inflammatory events include significant complement activation, neutrophil infiltration and significant cytokine secretions. The past literature suggests that complement activation may include a sex-linked protein. Therefore, we hypothesized that complement activation may vary by sex during intestinal ischemia/reperfusion (IR) injury. We subjected male and female mice to 30 min intestinal ischemia followed by 15, 30, 60 or 120 min reperfusion. Additional mice of each sex was subjected to sham surgery. Intestinal sections were stained and complement initiation pathways and time points of deposition determined. Complement regulatory molecules were also examined. Initial results suggest that males expressed more C1q compared to females, while females expressed more MBL than males. In addition, complement regulators varied by sex. Our data demonstrate that complement initiation and regulation differ between sexes during intestinal IR. These response differences in intestinal IR provide insight into the sexual dimorphism of ischemic intestine diseases and may aid in the development of sex-specific therapies that reduce intestinal injury.

Influence of Hyperglycemia During Different Phases of Ischemic Preconditioning on Cardioprotection-A Focus on Apoptosis and Aggregation of Granulocytes.

Ischemic preconditioning (IPC) protects the myocardium against ischemia/reperfusion injury. Evidence suggests that hyperglycemia inhibits IPC-induced cardioprotection. The effects of hyperglycemia initiated during different phases of IPC on myocardial injury were characterized with emphasis on apoptosis and aggregation of polymorphonuclear granulocytes (PMN). Male Wistar rats were subjected to 35 min of myocardial ischemia and 2 h of reperfusion. Control animals were not further treated. IPC was induced by three cycles of 3 min ischemia and 5 min of reperfusion before major ischemia. Hyperglycemia (blood glucose more than 22.2 mmol/L) was induced by glucose administration with or without IPC during different phases (trigger- (before ischemia), mediator- (during ischemia), early reperfusion-phase). One additional group received an anti-PMN-antibody before ischemia. Infarct size was quantified by triphenyltetrazolium chloride staining. Cytochrome C release and B-cell lymphoma two (Bcl-2) expression were assessed by western blot analysis. Poly-ADP-Ribose staining and PMN accumulation were quantified with immunohistochemistry and histochemistry. IPC reduced infarct size compared with control. Hyperglycemia completely abolished IPC-induced cardioprotection independent of the time point of initiation. Hyperglycemia before and during major ischemia but without IPC also slightly reduced infarct size. IPC reduced the accumulation of PMNs. This effect was reversed by hyperglycemia during trigger- and mediator-phase but not by hyperglycemia during reperfusion. Hyperglycemia alone had no effect on PMN accumulation. In all treatment groups, signs of myocardial apoptosis were reduced compared with control. IPC alone, combined with hyperglycemia and anti-PMN treatment, reduced apoptosis by a Bcl-2-associated mechanism. Hyperglycemia alone reduced apoptosis by a Bcl-2-independent pathway. Hyperglycemia inhibits IPC-induced cardioprotection independent of its onset. Furthermore, hyperglycemia prevents apoptosis and IPC-induced reduction of PMN aggregation.

Semi-Immersive Virtual Reality as a Tool to Improve Cognitive and Social Abilities in Preschool Children

Virtual reality (VR) creates computer-generated virtual environments where users can experience and interact in a similar way as they would do in real life. VR systems are increasingly being used for rehabilitation goals, mainly with adults, but also with children, extending their application to the educational field. This report concerns a study of the impact of a semi-immersive VR system in a group of 25 children in a kindergarten context. The children were involved in several different games and activity types, specifically developed with the aim of learning specific skills and foster team collaboration. Their reactions and behaviors were recorded by their teachers and by trained psychologists through observation grids addressing task comprehension, participation and enjoyment, interaction and cooperation, conflict, strategic behaviors, and adult-directed questions concerning the activity, the device or general help requests. The grids were compiled at the initial, intermediate and final timepoint during each session. The results show that the activities are easy to understand, enjoyable, and stimulate strategic behaviors, interaction and cooperation, while they do not elicit the need for many explanations. These results are discussed within a neuroconstructivist educational framework and the suitability of semi-immersive, virtual-reality-based activities for cognitive empowerment and rehabilitation purposes is discussed.

Early Inflammatory Signatures Predict Subsequent Cognition in Long-Term Virally Suppressed Women With HIV.

Immunologic function is an important determinant of cognition. Here we examined the contribution of early immune signatures to cognitive performance among HIV-infected, virally suppressed women (HIV+VS) and in HIV-uninfected (HIV-) women. Specifically, we measured serum inflammatory markers, developed combinatory immune signatures, and evaluated their associations with cognition. Forty-nine HIV+VS women in the Women’s Interagency HIV Study (WIHS) who achieved viral suppression shortly after effective antiretroviral therapy (ART) initiation, and 56 matched HIV− women were selected. Forty-two serum inflammatory markers were measured within 2 years of effective ART initiation for HIV+VS women, and at an initial timepoint for HIV− women. The same inflammatory markers were also measured approximately 1, 7, and 12 years later for all women. Of the 105 women with complete immune data, 83 (34 HIV+VS, 49 HIV−) also had cognitive data available 12 years later at ≥1 time points (median = 3.1). We searched for combinatory immune signatures by adapting a dynamic matrix factorization analytic method that builds upon Tucker decomposition followed by Ingenuity® Pathway Analysis to facilitate data interpretation. Seven combinatory immune signatures emerged based on the Frobenius residual. Three signatures were common between HIV+VS and HIV− women, while four signatures were unique. These inflammatory signatures predicted subsequent cognitive performance in both groups using mixed-effects modeling, but more domain-specific associations were significant in HIV+VS than HIV− women. Leukocyte influx into brain was a major contributor to cognitive function in HIV+VS women, while T cell exhaustion, inflammatory response indicative of depressive/psychiatric disorders, microglial activity, and cytokine signaling predicted both global and domain-specific performance for HIV− women. Our findings suggest that immune signatures may be useful diagnostic, prognostic, and immunotherapeutic targets predictive of subsequent cognitive performance. Importantly, they also provide insight into common and distinct inflammatory mechanisms underlying cognition in HIV− and HIV+VS women.

Time-series expression profiling of sugarcane leaves infected with Puccinia kuehnii reveals an ineffective defense system leading to susceptibility.

Successful orange rust development on sugarcane can potentially be explained as suppression of the plant immune system by the pathogen or delayed plant signaling to trigger defense responses. Puccinia kuehnii is an obligate biotrophic fungus that infects sugarcane leaves causing a disease called orange rust. It spread out to other countries resulting in reduction of crop yield since its first outbreak. One of the knowledge gaps of that pathosystem is to understand the molecular mechanisms altered in susceptible plants by this biotic stress. Here, we investigated the changes in temporal expression of transcripts in pathways associated with the immune system. To achieve this purpose, we used RNA-Seq to analyze infected leaf samples collected at five time points after inoculation. Differential expression analyses of adjacent time points revealed substantial changes at 12, 48h after inoculation and 12days after inoculation, coinciding with the events of spore germination, haustoria post-penetration and post-sporulation, respectively. During the first 24h, a lack of transcripts involved with resistance mechanisms was revealed by underrepresentation of hypersensitive and defense response related genes. However, two days after inoculation, upregulation of genes involved with immune response regulation provided evidence of some potential defense response. Events related to biotic stress responses were predominantly downregulated in the initial time points, but expression was later restored to basal levels. Genes involved in carbohydrate metabolism showed evidence of repression followed by upregulation, possibly to ensure the pathogen nutritional requirements were met. Our results support the hypothesis that P. kuehnii initially suppressed sugarcane genes involved in plant defense systems. Late overexpression of specific regulatory pathways also suggests the possibility of an inefficient recognition system by a susceptible sugarcane genotype.

Investigating the Range of Symptom Endorsement at Initiation of a Graduated Return-to-Play Protocol After Concussion and Duration of the Protocol: A Study From the National Collegiate Athletic Association–Department of Defense Concussion, Assessment, Research, and Education (CARE) Consortium

Background: Organizations recommend that athletes should be asymptomatic or symptom-limited before initiating a graduated return-to-play (GRTP) protocol after sports-related concussion, although asymptomatic or symptom-limited is not well-defined. Hypotheses: (1) There will be a range (ie, beyond zero as indicator of “symptom-free”) in symptom severity endorsement when athletes are deemed ready to initiate a GRTP protocol. (2) Baseline symptom severity scores and demographic/preinjury medical history factors influence symptom severity scores at the commencement of the GRTP protocol. (3) Greater symptom severity scores at GRTP protocol initiation will result in longer protocol duration. (4) Symptom severity scores will not differ between those who did and did not sustain a repeat injury within 90 days of their initial injury. Study Design: Cohort study; Level of evidence, 2. Methods: Across 30 universities, athletes (N = 1531) completed assessments at baseline and before beginning the GRTP protocol, as determined by local medical staff. Symptom severity scores were recorded with the symptom checklist of the Sport Concussion Assessment Tool–3rd Edition. Nonparametric comparisons were used to examine the effect of medical, demographic, and injury factors on symptom endorsement at GRTP protocol initiation, as well as differences in symptom severity scores between those who did and did not sustain a repeat injury within 90 days. A Cox regression was used to examine the association between symptom severity scores at GRTP protocol initiation and protocol duration. Results: Symptom severity scores at the time when the GRTP protocol was initiated were as follows: 0 to 5 (n = 1378; 90.0%), 6 to 10 (n = 76; 5.0%), 11 to 20 (n = 42; 3.0%), and ≥21 (n = 35; 2.0%). Demographic (sex and age), medical (psychiatric disorders, attention-deficit/hyperactivity disorder, learning disorder), and other factors (baseline symptom endorsement and sleep) were significantly associated with higher symptom severity scores at the GRTP initiation (P < .05). The 4 GRTP initiation time point symptom severity score groups did not significantly differ in total time to unrestricted RTP, χ2(3) = 1.4; P = .73. When days until the initiation of the GRTP protocol was included as a covariate, symptom severity scores between 11 and 20 (P = .02; hazard ratio = 1.44; 95% CI, 1.06-1.96) and ≥21 (P < .001; hazard ratio = 1.88; 95% CI, 1.34-2.63) were significantly associated with a longer GRTP protocol duration as compared with symptom severity scores between 0 and 5. Symptom severity scores at GRTP initiation did not significantly differ between those who sustained a repeat injury within 90 days and those who did not (U = 29,893.5; P = .75). Conclusion: A range of symptom severity endorsement was observed at GRTP protocol initiation, with higher endorsement among those with higher baseline symptom endorsement and select demographic and medical history factors. Findings suggest that initiation of a GRTP protocol before an absolute absence of all symptoms is not associated with longer progression of the GRTP protocol, although symptom severity scores >10 were associated with longer duration of a GRTP protocol. Results can be utilized to guide clinicians toward optimal GRTP initiation (ie, balancing active recovery with avoidance of premature return to activity).

Diabetic Nephropathy in Spontaneously Type 2 Diabetic Nonhuman Primates (Rhesus Monkeys Macaca mulatta): Comparator Validation of Angiotensin Receptor Blocker Valsartan

In patients, Type 2 Diabetes Mellitus (T2DM) and Diabetic Nephropathy (DN) have unknown initiation time points, likely well before their respective clinically‐defined features of elevated fasting glucose/HbA1c and reduced eGFR/microalbuminuria respectively, and have unknown preceding and/or causal mechanisms or events (genetic or otherwise). In part, this is due to animal models that do not fully reflect the human condition, do not show the same progressive processes preceding overt onset of clinically‐defined symptoms, and do not show similar associations of contributing factors including hypertension and cardiovascular disease (CVD)/metabolic syndrome (MetSyn). Nonhuman primates (NHPs, rhesus monkeys, Macaca mulatta) frequently, naturally and spontaneously develop obesity, MetSyn, T2DM and DN, with or without CVD, and in this study have been followed metabolically for diabetes‐related and cardiovascular disease features longitudinally from normal young adults to full expression of these diseases (colony N= 1198). We have identified the specific criteria for the identification of DN (eGFR CKD‐EPI 30–59 ml/min/1.73m2 or Iohexol clearance for direct GFR) and/or urine albumin/creatinine ratio (UACR ≥ 10 mg/g) and of CVD (cardiac function by ECHO HFpEF or HFrEF with LVEF &lt;50%, e’&lt;8 cm/s and E/e’ &gt;10) in these animals and have examined these features in relation to the progression of DN and T2DM (using ADA DM diagnostic criteria as applied to patients). Of those with CKD (N=52), 58% also had CVD. In addition, we report here the responses of adult monkeys, selected for the presence of T2DM and diabetic nephropathy ((increased eGFR and decreased urine albumin/creatinine ratio (UACR)) to the validation drug, an angiotensin receptor blocker, Valsartan. Monkeys (N=8 with both CKD and CVD) were studied before and at 2‐week intervals during the 8 week treatment period for the effects on blood pressure, Creatinine, Cystatin C, BUN, and eGFR. Significant improvements compared to vehicle were observed in eGFR (‐increased 23%), UACR (decreased 76%) and SBP (decreased14%) (all p’s &lt;0.05). There were no differences in cardiac ultrasound evaluations compared to vehicle. The similarities in all features and effects between the rhesus monkey and previous reports in humans in clinical settings are extraordinary. We have previously shown the histopathology of DN in monkeys to be “classical human glomerulopathy”, and show that these important similarities permit renal biopsies to be used at any desired intervals to follow changes in DN histopathology and molecular markers, and to evaluate their reversal or modification by therapeutic agents of direct relevance to human pathophysiology.Support or Funding InformationNIH National Institute on Aging: HHSN263200800022C; SCPrimed Shines Biotech Co., Ltd, Chengdu, Sichuan, China

Differential Regulation of DJ‐1 in Glial Cells upon MHV‐A59 ‐Induced Oxidative Stress

Astrocyte and microglial activation are two hallmarks associated with oxidative stress induction in neural cells. Generation of reactive oxygen species (ROS) is one of the molecular outcomes of neurotropic Mouse Hepatitis Virus A59 (MHV‐A59) infection in the Central Nervous System in an experimental animal model for human demyelinating disease Multiple Sclerosis. MHV‐A59 infection has been found to cause encephalitis, meningitis, and hepatitis in acute stage (Day 5–7 post‐infection) and progressive demyelination in chronic stage (Day 30 post‐infection). The virus has shown glial cell tropism leading to its activation, although the role of oxidative stress in this aspect is not well understood. Therefore, our work focuses on the regulation and function of important anti‐oxidative markers like DJ‐1, Nrf2, and HMOX‐1, upon viral infection in animal CNS tissue, isolated cells from tissues, primary culture and cell culture‐based studies. All three genes were found to be upregulated in vivo in mice brain at acute stage (Day 3 and 5) and chronic stage (Day 30) post‐infection. Similar upregulation was also observed in isolated microglia from mice brain, primary astrocytes, and microglia cultured from neonatal mice brain. In vitro studies with DBT astrocytoma cell line and N9 microglia cell line showed oxidative stress generation and upregulation in all three genes at initial time points of viral infection; however all the genes were found to be downregulated at higher time points. DBT cells exhibited decrease in cell viability and oxidative stress generation upon viral infection, and such changes had been observed to be ameliorated when DJ‐1 was stably transfected into DBT cells. Literature studies indicate that XBP1, an ER stress marker, upregulates DJ‐1 expression upon oxidative stress induction. In silico and in vitro studies showed that XBP1 was differentially activated in both DBT and N9 cells and was associated with DJ‐1 promoter upon viral infection in N9 cells. MHV‐A59 has shown tropism for glial cells resulting in their activation, and oxidative stress‐related cellular mechanisms involving these genes can contribute to the activation process of these glial cells.Support or Funding InformationThis study was supported by the Department of Biotechnology (DBT) and Indian Institute of Science Education and Research Kolkata (IISER Kolkata). Soumya Kundu was supported by University Grant Commission (UGC). Gisha Rose Anthony, Vineeth A. Raveendran, and Rahul Kumar were supported by DST‐INSPIRE fellowship under the banner of IISER Kolkata.

Predicting limiting ‘free sugar’ consumption using an integrated model of health behavior

Excess intake of ‘free sugars’ is a key predictor of chronic disease, obesity, and dental ill health. Given the importance of determining modifiable predictors of free sugar-related dietary behaviors, we applied the integrated behavior change model to predict free sugar limiting behaviors. The model includes constructs representing ‘reasoned’ or deliberative processes that lead to action (e.g., social cognition constructs, intentions), and constructs representing ‘non-conscious’ or implicit processes (e.g., implicit attitudes, behavioral automaticity) as predictors of behavior. Undergraduate students (N = 205) completed measures of autonomous and controlled motivation, the theory of planned behavior (TPB) measures of explicit attitude, subjective norms, perceived behavioral control (PBC), and intentions, past behavior, implicit attitude, and behavioral automaticity at an initial point in time, and free sugar limiting behavior and behavioral automaticity two weeks later. A Bayesian structural equation model indicated that explicit attitude, subjective norms, and PBC predicted behavior via intention. Autonomous motivation predicted behavior indirectly through all TPB variables, while controlled motivation predicted behavior only via subjective norms. Implicit attitudes and behavioral automaticity predicted behavior directly and independently. Past behavior predicted behavior directly and indirectly through behavioral automaticity and intentions, but not implicit attitudes. Current findings suggest pervasive effects of constructs representing both reasoned and non-conscious processes and signpost potential targets for behavioral interventions aimed at minimizing free sugar consumption.

Serum albumin changes and mortality risk of peritoneal dialysis patients.

Initial or single time point serum albumin levels have been shown to be important in predicting the prognosis of peritoneal dialysis (PD) patients. However, we assume the dynamic change and trend of albumin after PD are essential. We aimed to investigate the association between baseline albumin levels, albumin trajectories, and patient mortality in a retrospective cohort study. In this retrospective cohort study, 547 incident PD patients were enrolled from Peking University Third Hospital. Date were collected by medical records review, including age, gender, body mass index, primary disease, comorbidities, and laboratory tests. A joint model for longitudinal data and time-to-event data was used to establish the relationship between serum albumin trajectories and mortality risk of PD patients. The albumin trajectories was negatively correlated with risk of death. The increase in the current value of albumin trajectories at time points t after PD was associated with decreased risk of death (HR = 0.881, p < 0.0001). There was no statistical association between initial albumin and risk of death (HR = 1.030, 95% CI 0. 995-1.066). The results showed that increased age, higher albumin-corrected Ca levels, and higher eGFR values were risk factors for death. In addition, predictors of low albumin levels are increased PD time, increased age, increased albumin-corrected Ca, and decreased BMI as well as initial albumin levels. This study demonstrates that albumin trajectories after PD is better than initial serum albumin level in predicting mortality risk. Increasing albumin level over time can improve the prognosis of PD patients.

Acute Oral Toxicity Evaluation of Ethyl Acetate Fraction of Sesbania Grandiflora Linn. Roots as Per OECD 425 TG on Albino Mice

Objective To evaluate the acute oral toxicity of ethyl acetate fraction of Sesbania grandiflora Linn. roots as per OECD 425 guidelines. Methods The two groups of five female albino mice were selected and both the groups of mice were observed for 14 days for various parameters according to OECD guidelines 425 guidelines after treating one group as a normal control and other with ethyl acetate fraction of Sesbania grandiflora root EFSGR 2000mgkg p.o.Results Oral dose of EFSGR 2000mgkg did not resulted in death and treatment related adverse effects such as modification in food and water consumption body weight variation relative organ weights biochemical and hematological parameters. But treatment showed mild changes in behavioral patterns like sleeping and itching during initial time point. Further histopathological evaluation of vital organs exhibited normal architecture and this observation provided good support to general and biochemical measurements.Conclusion The current study showed that the ethyl acetate fraction of Sesbania grandiflora did not exhibit any signs of toxicity in female albino mice. Hence proved that the toxicity study based calculated dose of EFSGR can be used for the pharmacological studies.

Reasoned and implicit processes in heavy episodic drinking: An integrated dual-process model.

Objectives University students commonly engage in heavy episodic drinking (HED), which contributes to injury risk, deleterious educational outcomes, and economic costs. Identification of the determinants of this risky behaviour may provide formative evidence on which to base effective interventions to curb HED in this population. Drawing from theories of social cognition and dual-process models, this study tested key hypotheses relating to reasoned and implicit pathways to action for HED in a sample of Australian university students who drink alcohol. Design A two-wave correlational design was adopted. Methods Students (N=204) completed self-reported constructs from social cognition theories with respect to HED at an initial time point (T1): attitude, subjective norm, perceived behavioural control, intentions, habit, past behaviour, and implicit alcohol identity. Four weeks later (T2), students self-reported their HED behaviour and habit. Results An initial path model indicated attitude and subjective norm predicted intentions, and intentions and implicit alcohol identity predicted HED. Inclusion of past behaviour and habit revealed direct effects of these on HED. Effects of T1 habit on HED were indirect through T2 habit, and there were indirect effects of past behaviour on HED through habit at both time points and the social cognition constructs. Direct effects of intentions and implicit alcohol identity, and indirect effects of attitude and subjective norm, on HED, were attenuated by the inclusion of past behaviour and habit. Conclusion Results indicate that university students' HED tends to be governed by non-conscious, automatic processes than conscious, intentional processes. Statement of contribution What is already known on this subject? Social cognitive factors are associated with risky alcohol consumption behaviours. Dual-process models are being used to explain health behaviours, such as heavy episodic drinking (HED). What does this study add? Past HED behaviour and HED habits have direct and indirect effects on students' HED behaviour. Past behaviour and habit attenuate the effects of intentions and implicit alcohol identity on HED.

Are There Sex-Specific Neuromuscular or Force Responses to Fatiguing Isometric Muscle Actions Anchored to a High Perceptual Intensity?

Keller, JL, Housh, TJ, Hill, EC, Smith, CM, Schmidt, RJ, and Johnson, GO. Are there sex-specific neuromuscular or force responses to fatiguing isometric muscle actions anchored to a high perceptual intensity? J Strength Cond Res 36(1): 156-161, 2022-The purpose of this study was to use the ratings of perceived exertion (RPE) clamp model to examine sex-specific changes in neuromuscular responses and force after a sustained isometric leg extension muscle action anchored to RPE = 8. Twenty adults (10 men and 10 women) performed sustained, isometric leg extension muscle actions at RPE = 8. Electromyographic (EMG) and mechanomyographic signals were recorded from the dominant leg. Neuromuscular and force values resulting from the sustained muscle action were normalized to pretest maximal voluntary isometric contractions (MVICs). The level of significance set for the study was p ≤ 0.05. The pretest MVIC was significantly (p < 0.001) greater (averaged across sex) than posttest MVIC force (55.5 ± 10.0 vs. 47.6 ± 11.1 kg). There was a significant (p < 0.01) decrease from pretest (95.4 ± 7.7 Hz) to posttest (76.2 ± 5.9 Hz) in EMG mean power frequency (MPF) for the men. The normalized force (averaged across sex) decreased significantly (p < 0.001) from the initial timepoint (57.1 ± 16.4%) to the final timepoint (44.3 ± 15.7%) of the sustained muscle action. Normalized EMG MPF (averaged across sex) decreased significantly (p = 0.001) from the initial timepoint (96.4 ± 17.5%) to final timepoint (87.8 ± 18.1%). The men and women exhibited similar fatigue-induced changes in force and neuromuscular parameters; therefore, these findings did not indicate different sex-specific responses after the fatiguing task anchored to a high perception of exertion. The force corresponding to RPE = 8 did not match the anticipated value; so, RPE and percentages of MVIC cannot be used interchangeably, and sustained isometric muscle actions anchored to RPE may elicit unique neuromuscular adaptations.

Eligibility of sodium-glucose co-transporter-2 inhibitors among patients with diabetes mellitus admitted for heart failure.

AimsSodium–glucose co‐transporter (SGLT)‐2 inhibitors have been shown to reduce the risk of cardiovascular death and heart failure (HF) hospitalization in patients with type 2 diabetes mellitus (DM) and high cardiovascular risk in two large clinical outcome trials: empagliflozin in EMPA‐REG OUTCOME and canagliflozin in CANVAS. The scope of eligibility for SGLT‐2 inhibitors (empagliflozin and canagliflozin) among patients with type 2 DM and HF, based on clinical trial criteria and current US Food and Drug Administration (FDA) labelling criteria, remains unknown.Methods and resultsUsing data from the US Get With The Guidelines (GWTG)—Heart Failure registry, we evaluated the proportion of patients with DM and HF eligible for SGLT‐2 inhibitor therapy based on the clinical trial criteria and the US FDA labelling criteria. The GWTG‐HF registry is a quality improvement registry of patients admitted in hospital with HF in the USA. We included GWTG‐HF registry participants meeting eligibility criteria hospitalized between August 2014 and 30 June 2017 from sites fully participating in the registry. The initial inclusion time point reflects when both drugs had FDA approval. Among the 139 317 patients (out of 407 317) with DM hospitalized with HF (in 460 hospitals; 2014 to 2017), the median age was 71 years, 47% (n = 65 685) were female, and 43% (n = 59 973) had HF with reduced ejection fraction. Overall, 43% (n = 59 943) were eligible for the EMPA‐REG OUTCOME trial, 45% (n = 62 818) were eligible for the CANVAS trial, and 34% (n = 47 747) of patients were eligible for either SGLT‐2 inhibitors based on the FDA labelling criteria. Among the FDA‐eligible patients, 91.5% (n = 43 708) were eligible for either the EMPA‐REG OUTCOME trial or the CANVAS trial. Patients who were FDA eligible, compared with those who were not, were younger (70.0 vs. 72.0 years of age), more likely to be male (57.7 vs. 50.3%), and had less burden of co‐morbidities.ConclusionsThe majority of patients with DM who are hospitalized with HF are not eligible for SGLT‐2 inhibitor therapies. Ongoing studies evaluating the safety and efficacy of SGLT‐2 inhibitors among patients with HF may potentially broaden the population that may benefit from these therapies.

INOS dependent and independent phases of lymph node expansion in mice with TNF-induced inflammatory-erosive arthritis

IntroductionA pivotal effect of lymphatic vessel (LV) function in joint homeostasis was identified in the tumor necrosis factor-transgenic (TNF-Tg) mouse model of rheumatoid arthritis (RA). Specifically, loss of LV contractions is associated with progressive synovitis and erosions. Furthermore, draining lymph node expansion is a biomarker of arthritic progression, and both macrophages and lymphatic endothelial cells express inducible nitric oxide synthase (iNOS), which disrupts LV contraction and transport of immune cells to the draining lymph nodes. Therefore, to directly assess these relationships, we tested the hypothesis that TNF-Tg mice with global genetic ablation of iNOS (iNOS−/−) will show delayed draining lymph node expansion, maintained LV contractions, and decreased synovitis and erosions.MethodiNOS−/−× TNF-Tg female and male mice, and control littermates (iNOS−/−, TNF-Tg, and WT), were examined with (1) ultrasound to determine popliteal lymph node (PLN) volume and (2) near-infrared imaging (NIR) to assess popliteal LV contraction frequency, and differences between genotypes were assessed at 3, 4, 5, and 6 months of age. Knees and PLN were harvested at 4 months in females and 6 months in males, to assess synovitis, bone erosions, and cellular accumulation in PLN sinuses via histology.ResultsInitially, an increase in PLN volume was observed for both female and male iNOS−/−× TNF-Tg and TNF-Tg compared to their WT and iNOS−/− counterparts at 2 and 3 months, respectively. Subsequently, TNF-Tg PLNs continue to increase in volume, while iNOS−/−× TNF-Tg did not increase in volume from the initial timepoints. WT and iNOS−/− PLN volume was unchanged throughout the experiment. LV contraction frequency was increased at 4 months in females and 5 months in males, in the iNOS−/−× TNF-Tg mice compared to the TNF-Tg. Synovitis and erosions were moderately reduced in iNOS−/−× TNF-Tg versus TNF-Tg knees in females, while no differences in knee pathology were observed in males.ConclusionsGenetic iNOS ablation maintains draining lymph node volume and LV function during TNF-induced inflammatory arthritis and is associated with moderately decreased joint inflammation and damage.

Analysis of the Sub-Clonal Structure of Smoldering Myeloma over Time Provides a New Means of Disease Monitoring and Highlights Evolutionary Trajectories Leading to Myeloma

Analysis of the Sub-Clonal Structure of Smoldering Myeloma over Time Provides a New Means of Disease Monitoring and Highlights Evolutionary Trajectories Leading to Myeloma

Expression Analysis of mRNA Decay of Maternal Genes during Bombyx mori Maternal-to-Zygotic Transition.

Maternal genes play an important role in the early embryonic development of the silkworm. Early embryonic development without new transcription depends on maternal components stored in the egg during oocyte maturation. The maternal-to-zygotic transition (MZT) is a tightly regulated process that includes maternal mRNAs elimination and zygotic transcription initiation. This process has been extensively studied within model species. Each model organism has a unique pattern of maternal transcriptional clearance classes in MZT. In this study, we identified 66 maternal genes through bioinformatics analysis and expression analysis in the eggs of silkworm virgin moths (Bombyx mori). All 66 maternal genes were expressed in vitellogenesis in day eight female pupae. During MZT, the degradation of maternal gene mRNAs could be divided into three clusters. We found that eight maternal genes of cluster 1 remained stable from 0 to 3.0 h, 17 maternal genes of cluster 2 were significantly decayed from 0.5 to 1.0 h and 41 maternal genes of cluster 3 were significantly decayed after 1.5 h. Therefore, the initial time-point of degradation of cluster 2 was earlier than that of cluster 3. The maternal gene mRNAs decay of clusters 2 and 3 is first initiated by maternal degradation activity. Our study expands upon the identification of silkworm maternal genes and provides a perspective for further research of the embryo development in Bombyx mori.

A Health Education Intervention to Increase Fiber Intake: A Randomized Clinical Trial

The intake level of fiber, a nutrient with a vast number of health benefits, falls behind the recommendations nationwide. The impact of nutrition education on improving daily intake has thus been investigated and shown to be effective in enhancing the intake of specific fiber-rich foods. Not much has been done to probe the effect of education on enhancing fiber intake with emphasis on all sources of dietary fiber. Thus, we conducted a randomized controlled trial to determine if a multi-component intervention—the Full Plate Diet— that places emphasis on fiberrich foods, impacts the dietary fiber intake to a level that exceeds the recommendations of the Institute of Medicine. Thirty-five subjects (7 males and 28 females) were randomly assigned to either the intervention group (n=20), which received the nutrition/health education or to the control group (n=15) which did not receive the education. A collective analysis of the two groups’ dietary intake was divided into three time points; April, May, and June. The recalled intake of dietary fiber and food groups considered to be rich sources of dietary fiber as well as macronutrients were averaged for each of these time points for each treatment group. For the purpose of comparing changes in intake of dietary fiber across the 3 time points and between the intervention and control groups, a linear mixed-model was used. The delta changes between the various time points were computed by subtracting the final time point minus the initial time point. Results showed that fiber intake increased significantly by 9.3 grams from the first time-point (April) to the second time-point (May) (95% CI 2.2, 16.4). The intake of legumes significantly improved by 0.36 (95% CI: 0.01, 0.7) and by 0.65 (95% CI: 0.3, 1.0) servings in the intervention group between baseline and the second and third-time (June) points, respectively, but did not change in the control group. Short-term education with emphasis on fiber intake from various food sources could improve the daily fiber intake. Longer follow-up investigations should also be conducted to assess whether or not health education could significantly improve the level of fiber on a longterm basis.

Regulation of Osteogenic Markers at Late Stage of Osteoblast Differentiation in Silicon and Zinc Doped Porous TCP.

Calcium phosphates (CaPs) are one of the most widely used synthetic materials for bone grafting applications in the orthopedic industry. Recent trends in synthetic bone graft applications have shifted towards the incorporation of metal trace elements that extend the performance of CaPs to have osteoinductive properties. The objective of this study is to investigate the effects of silicon (Si) and zinc (Zn) dopants in highly porous tricalcium phosphate (TCP) scaffolds on late-stage osteoblast cell differentiation markers. In this study, an oil emulsion method is utilized to fabricate highly porous SiO2 doped β-TCP (Si-TCP) and ZnO doped β-TCP (Zn-TCP) scaffolds through the incorporation of 0.5 wt.% SiO2 and 0.25 wt.% ZnO, respectively, to the β-TCP scaffold. Reverse transcriptase quantitative polymerase chain reaction (RT-qPCR) is utilized to analyze the mRNA expression of osteoprotegerin (OPG), receptor activator of nuclear kappa beta ligand (RANKL), bone morphogenetic protein 2 (BMP2), and runt-related transcription factor 2 (Runx2) at the later stage of osteoblast differentiation, day 21 and day 28. Results show that the addition of Si and Zn to the β-TCP structure inhibited the β to α-TCP phase transformation and enhance the density without affecting the dissolution properties. Normal BMP-2 and Runx2 transcriptions are observed in both Si-TCP and Zn-TCP scaffolds at the initial time point, as demonstrated by RT-qPCR. Moreover, the addition of both Si and Zn positively regulate the osteoprotegerin: receptor activator of nuclear factor k-β ligand (OPG:RANKL) ratio at 21-days for Si-TCP and Zn-TCP scaffolds. These results demonstrate the effects of Si and Zn doped porous β-TCP scaffolds on the upregulation of osteoblast marker gene expression including OPG, RANKL, BMP-2, and Runx2, indicating the role of trace elements on the effective regulation of late-stage osteoblast cell differentiation markers.