bluuebunny/crossref_metadata_2025_split huggingface dataset:
An exploration of functional domains in the zebrafish ortholog of human Usher syndrome gene USH2A
Authors: Drake Rachel Rae , Phillips Jennifer , Westerfield Monte 
Published: June 2021

Usher syndrome is the most prevalent cause of hereditary deafblindness. Usher type 2A is the most widespread form of the syndrome, caused by mutations in the USH2A gene. USH2A patients experience congenital hearing impairment due to defects in the mechanosensory hair cells of the inner ear and progressive retinal degeneration. USH2A consists of 72 exons encoding the protein Usherin. Several functional domains have been identified and human mutations causing Usher Syndrome have been located in multiple regions of the protein. Determining the relative importance of these functional domains is crucial to the development of effective therapies.We designed morpholino oligonucleotides to interfere with pre‐mRNA splicing at the 5′ and 3′ regions of the transcript. Protein levels in sensory cells were visualized with antibody staining. Both morpholinos achieved a similar degree of Usherin depletion, suggesting that altered splicing at either the 5′ or 3′ region can impact Usherin production or stability. We noted circle‐swimming and failure to inflate the swim bladder in morphant larvae, both common phenotypes indicating hair cell defects. Because of the retinal degeneration seen in human Usher syndrome, we analyzed retinal cells in morphants and saw significant increases in photoreceptor death. These results will enable further functional analyses of zebrafish Usher proteins.
Validation of Toddler NutriSTEP®
Authors: Simpson Janis Randall , Whyte Kylie , Lac Jane , Rysdale Lee , Keller Heather , Beyers Joanne 
Published: June 2021

The objective was to establish criterion and construct validity for Toddler NutriSTEP®, a 17‐item, community‐based parent‐administered nutrition risk screening questionnaire (total score = 68), adapted from the original preschooler version. Parents of 200 toddlers (18–35 mo), recruited from community settings, completed Toddler NutriSTEP® and a demographic questionnaire; an assessment by a registered dietitian (RD) included medical and nutritional history, weight, height, 3‐d food records. The RD rated nutritional risk on a scale of 1 (low) to 10 (high). Receiver operating curves (ROC) were used to determine sensitivity and specificity for various cut‐points on the scale. Differences in Toddler NutriSTEP® scores for selected demographic variables were determined by ANOVA.Scores on Toddler NutriSTEP® and the RD rating were correlated (r=0.66, p<0.000). Areas under the ROC curves for the moderate risk (score 5–7) and the high risk (score 8+) RD ratings were 0.87 and 0.82 with cut points of >;20 & >;25, respectively. Scores on Toddler NutriSTEP® were significantly higher (more risk) for parents with low incomes, low education and for immigrants to Canada.The Toddler NutriSTEP® questionnaire demonstrates both criterion and construct validity in children 18 – 35 months. Funding: Canadian Institutes of Health Research
Early Intervention and Reduction of Patient Default Improve Cost Effectiveness of a Supplementary Feeding Intervention for HIV+ Malnourished Patients in Ethiopia
Authors: Rogers Beatrice Lorge , Coates Jennifer , Passarelli Simone , Bontrager Elizabeth , Suri Devika , Ghosh Shibani , Kidane Yitbarek , Sadler Kate 
Published: June 2021

Objectiveassess marginal cost and cost‐effectiveness of adding a nutritional supplement to standard treatment of wasted HIV+ patients to raise BMI.MethodsSupplement was provided in 8 clinics; 8 non‐randomly assigned comparison clinics provided standard care: ART, counseling. Costs included: supplement, staff time; administrative cost. Patients’ time in clinic visits was tracked.ResultsRecovery was defined as BMI=18.5 for 2 visits. Treatment patients spent more time per visit. Marginal cost per recovered patient was $12,192 for those entering with severe acute malnutrition (SAM=BMI<= 16) and $1980 for those with moderate acute malnutrition (MAM=BMI >; 16 < 18.5) due to low recovery in SAM patients. Default (failure to complete treatment) was high (70.6% treatment; 59.6% comparison). Three models assessed potential effects of reducing default. Reducing default substantially increased recovery rates and reduced marginal cost per recovered patient; costs were much lower for patients entering as MAM than SAM.ConclusionsSupplementation of wasted HIV+ patients should start early for maximum cost‐effectiveness; reducing default greatly increases cost‐effectiveness.No conflict of interest.
Prevalence of Vitamin A Deficiency and Associations with Socioeconomic Status in Lactating HIV‐infected Malawian Mothers
Authors: Patel K. D. , Bentley M. E. , Flax V. L. , Shahab‐Ferdows S. , Allen L. H. , Chasela C. S. , Kayira D. , Tegha G. , Jamieson D. J. , van der Horst C. M. , Adair L. S. 
Published: June 2021

Vitamin A deficiency (VAD) is a concern in HIV+ women because it lowers epithelial integrity, the immune response, and red blood cell production, leading to greater risk of negative health outcomes. A subsample of Malawian lactating HIV+ mothers enrolled in the Breastfeeding, Antiretrovirals, and Nutrition Study, with demographic and plasma retinol binding protein (RBP) data (n=623), was evaluated to understand prevalence of VAD (serum RBP ≤1.17 μmol/L) and the correlation between socioeconomic status (SES) and VAD. The Thurnham method was used to adjust RBP values for inflammation, using C‐reactive protein>;5 mg/L and α‐1‐acid glycoprotein>;1 g/L. The prevalence of VAD was 71% at 2 or 6 weeks postpartum and 77% at 24 weeks postpartum and did not change when adjusted for inflammation. Logistic regression models were used to calculate odds of deficiency for each SES indicator. Lower maternal age and having no paid work were associated with higher levels of VAD (p<0.05), but VAD was unrelated to other SES indicators. The high level of VAD in this population, especially among younger and poorer mothers, suggests the need for vitamin A supplementation, but concerns that supplementation is associated with increased viral shedding and a higher risk of mother‐to‐child HIV transmission need to be weighed in settings with high HIV prevalence.Funding: CDC(U48‐DP000059–01); Gates Foundation(OPP53107)
Resistant Starch Promotes Vitamin D Balance in Type 1 Diabetic Mice
Authors: Anderegg Alysse S. , Schalinske Kevin L. 
Published: June 2021

Type 1 diabetes mellitus (T1DM) is a chronic disease characterized by abnormal carbohydrate metabolism and hyperglycemia owing to beta cell dysfunction and insulin deficiency. A serious complication of T1DM is diabetic nephropathy, which can result in vitamin D imbalance, characterized by the loss of 25‐OH‐vitamin D in the urine. Using streptozotocin (STZ)‐treated rats as a model of T1DM, we have demonstrated that cooked dietary resistant starch (RS) attenuates the weight loss associated with T1DM and prevents the loss of vitamin D binding protein and 25‐OH‐vitamin D in the urine. The focus of this study was to determine if uncooked dietary RS would produce similar effects in a STZ‐induced mouse model of T1DM. In this study, 18 female mice were randomly assigned to one of 3 treatment groups: control, T1DM, and T1DM + uncooked dietary RS. Control and T1DM mice were fed a standard cornstarch diet during the 6‐wk treatment period, whereas T1DM + RS group were fed a diet containing high amylose cornstarch (36% resistant to digestion). To date, we have observed similar results in the mouse model, including diminished weight loss owing to the positive effect of dietary RS. Future research will be directed toward understanding the mechanism underlying the impact of RS on maintaining vitamin D balance, owing to the potential prevention of diabetic nephropathy in type 1 diabetic mice.Support: Plant Sciences Institute, ISU.Grant Funding Source: Plant Sciences Institute, ISU
Chronic alcohol consumption has greater impact on hepatic DNA hydroxymethylation in young mice relative to old
Authors: Tammen Stephanie A. , Dolnikowski Gregory G. , Ausman Lynne M. , Sauer Julia , Friso Simonetta , Choi Sang‐Woon 
Published: June 2021

Aging and chronic alcohol consumption affects DNA methylation, changing gene transcription. It is not yet known whether these factors also alter DNA hydroxymethylation (hmC), an epigenetic mark produced by oxidation of methylcytosine. Our objective was to determine changes in hmC caused by aging and alcohol. Young (4 months; n=20) and old (18 months; n=18) male C57BL/6 mice were pair‐fed a Lieber‐DeCarli liquid diet with or without alcohol (18% of energy) for 5 weeks. Global hmC was analyzed from hepatic DNA using an LC/MS‐MS method and calculated as percent of total cytosine. Differentially hydroxymethylated regions (DhMRs) were determined by microarray‐based hydroxymethyl‐DNA immunoprecipitation (hmeDIP‐chip). Relative to the controls, chronic alcohol consumption did not alter global hmC in old mice, but significantly reduced global hmC in young mice (0.22%±0.01% vs 0.29±0.06%, p=0.004). Old mice had significantly lower global DNA hmC than young mice (0.24±0.02% vs 0.29±0.06%, p=0.04). This model also suggests an interaction between aging and alcohol in determining global hmC levels (pint.=0.01). Genomic hmC patterns followed a similar trend; the alcohol diet resulted in more DhMRs in young mice compared to old (431 vs 196 DhMRs). This is the first study on the effects of alcohol consumption and aging on hmC levels and posits that chronic alcohol consumption differently affects hmC in young and old mice.Grant Funding Source: NIH grant R01 AG025834
The Influence of Type II Diabetes on the Immune Response of Obese Adults
Authors: Paich Heather A. , Sheridan Patricia A. , Noah Terry L. , Weir Samuel S. , Beck Melinda A. 
Published: June 2021

We have previously shown that obesity impairs expression of CD4+ and CD8+ T cell markers of activation and function. Obesity is associated with type II diabetes; however, not all obese individuals develop type II diabetes. Type II diabetes is associated with low levels of chronic inflammation and a higher prevalence of infections. The objective of our study was to elucidate the differences in the human cellular immune response between obese individuals with type II diabetes and obese individuals without type II diabetes. We stimulated peripheral blood mononuclear cells, obtained from obese individuals with and without type II diabetes, ex vivo with live pandemic influenza virus A and anti‐CD3/anti‐CD28 beads. Markers of activation, function, and signaling were measured using flow cytometry. We also measured three parameters of cellular glycolysis, including glycolytic rate, glycolytic capacity, and glycolytic reserve capacity, in T cells stimulated with or without anti‐CD3/anti‐CD28 bead stimulation and with or without insulin stimulation. We found significant differences between obese individuals with and without type II diabetes, demonstrating that metabolic disruptions in insulin signaling further increase immune dysfunction in obese adults.Grant Funding Source: NIH ROI AI078090 and P30DK056350
CYTOKINE PROFILE IN CELIAC DISEASE: A LASER CAPTURE MICRODISSECTION APPROACH
Authors: Iaquinto Gaetano , Aufiero Vera Rotondi , Preziuso Marco , Marano Angela , Stillitano Ilaria , Giardullo Nicola , Bevilacqua Rosa Maria , Iacomino Pino , Mazzarella Giuseppe 
Published: June 2021

BackgroundDifferent approaches have been used to study the pattern of cytokines in celiac disease (CD), which have led sometimes to contradictory results. Laser capture microdissection (LCM) is a powerful tool for the isolation of specific cell populations from stained sections.AimCharacterize the pattern of cytokines within the tissue compartments of the small intestinal mucosa of CD patients, dissected by LCM.MethodsFrozen sections of jejunal biopsies were obtained from 7 untreated CD patients. Epithelium and lamina propria were isolated separately by LCM, and the mRNA levels for IFN‐γ, IL‐21, IL‐15, IL‐10 and TGF‐β, were determined by using real‐time PCR.ResultsWe observed a higher expression of IFN‐γ and IL‐21 mRNA in the lamina propria than in the epithelium and a higher expression of IL‐15 mRNA in the epithelium than in the lamina propria. Same levels of IL‐10 and TGF‐β transcripts were detected in both the compartments of untreated celiac mucosa.ConclusionOur data suggest that the epithelium produce cytokines involved in the innate immunity while lamina propria cells produce cytokines involved in the adaptive immune response. Both epithelial and lamina propria compartments produce immunosuppressive cytokines. This work underlines the importance of LCM as a valuable tool to determine inflammatory components in specific intestinal compartments enhancing understanding of CD pathogenesis.
Omega‐3 derived Resolvin D1 reduces the severity of inflammatory arthritis
Authors: Norling Lucy Victoria , Perretti Mauro 
Published: June 2021

Dietary supplementation with omega‐3 PUFA is efficacious in reducing joint pain, morning stiffness and NSAID usage in Rheumatoid arthritis patients. However, the mechanisms by which omega‐3 PUFA exert their beneficial effects has not been fully explored. Recent discoveries have alluded that omega‐3 PUFA can be enzymatically converted in vivo to novel bioactive lipid mediators, which promote the resolution of inflammation and are log‐orders more potent than their precursors, termed resolvins. We recently showed that resolvin D1 (RvD1) limits PMN recruitment in experimental peritonitis via its receptor Fpr2, yet whether RvD1 exhibits anti‐arthritic effects remains to be elucidated. We performed the K/BxN serum transfer model of arthritis (50μl serum i.p. on day 0 and day 2) on mice fed control or 10% salmon oil‐enriched diet, in comparison with mice administered daily 17R‐RvD1 (100ng, i.p.) or vehicle (0.1% EtOH), and graded arthritis severity (max. 12). Omega‐3 enriched diet mildly reduced disease severity, yet RvD1‐treated mice exhibited a significantly lower clinical score, and reduced hind paw oedema compared to vehicle‐treated mice. These studies indicate that the endogenous pro‐resolving mediator RvD1 is more efficacious than an omega‐3 enriched diet at reducing the severity of inflammatory arthritis and that omega‐3 PUFA and derivatives are a viable approach to novel anti‐arthritic therapies.Supported by Arthritis Research UK Fellowship 19909 to LVN.
Therapeutic use of eNOS/Caveolin‐1 antagonistic peptides for endothelial dysfunction and atherogenesis
Authors: Sharma Arpeeta , Leung Cleo , Stefanovic Nada , de Haan Judy , Bernatchez Pascal 
Published: June 2021

RationaleEndothelial dysfunction, which is characterized by a reduction in nitric oxide (NO) bioavailability, is an early pro‐atherogenic step. Endothelial nitric oxide synthase (eNOS), the enzyme responsible for the constitutive production of NO, is negatively regulated by its association to Caveolin‐1 (Cav‐1), the major coat protein of caveolae. Whether antagonizing the eNOS/Cav‐1 interaction is an anti‐atherosclerotic drug target is unknown.ObjectiveWe have recently shown that a mutant Cav‐1 derived peptide (CavNoxin) can increase NO release by antagonizing eNOS binding to Cav‐1 (Bernatchez, Sharma et al., JCI 2011). Since upregulating NO production is therapeutically relevant in atherosclerosis we hypothesized that CavNoxin can attenuate atherosclerotic progression in vivo.Methods & ResultsApoE knockout mice were placed on a high fat diet for 12 weeks and were injected with either CavNoxin (2.5mg/kg) or vehicle peptide every 3 days. At the end of the 12 weeks, atherosclerotic lesions were analysed. CavNoxin treatment attenuated atherosclerotic lesions in the aorta and aortic sinuses by 42% and 21% respectively, as compared to vehicle controls. In addition, Cavnoxin reduced VCAM‐1 (adhesion molecule) expression and oxidative stress (measured by DHE and nitrotyrosine staining) in vivo.ConclusionThese data are the first to show an effect of the eNOS/Cav‐1 antagonism in atherosclerosis.
Sex‐specific endothelial cell response to insulin
Authors: Bingaman Susan , Huxley Virginia H. 
Published: June 2021

We have observed that following acute exposure to insulin, permeability of mesenteric microvessels to albumin (Ps) differs by sex (S Bingaman et al., FASEB J20 406.7 2006; R Sasaki et al., FASEB J20 205.3 2006).Occupancy of the endothelial cell (EC) insulin receptor leads to the activation of 2 pathways that generate the vasodilator nitric oxide on the one hand and the vasoconstrictor endothelin‐1 on the other. Given our working hypothesis that agents that are vasodilators elevate Ps, and agents that are vasoconstrictors reduce Ps, we posited that the sex‐specific responses to insulin result from sex differences in the contributions of the 2 pathways.To evaluate the cell signaling EC isolated from the mesenteries of adult male and female Sprague Dawley rats were grown in culture for up to 7 passages. Cell‐based ELISA was used to assess total and phosphorylated endothelial nitric oxide synthase (eNOS) and MAPK, and insulin receptor‐β (IRβ) expression after 30 minutes exposure to 10−7M Insulin as compared to its absence. Results were normalized against β‐tubulin expression for each group.IRβ expression was reduced only in EC from males, while both sexes showed differences in eNOS and MAPK expression and activity. These data demonstrate the capacity for vascular cells to retain and display sex‐differences in cell signaling in the absence of sex hormones.Support: HL093068, HCO6RR017353 & the MU Pulmonary/MPP Partnership
Relapse of clinical symptoms in the acute experimental autoimmune encephalomyelitis (EAE) is adrenalectomy (ADX) time dependent in the Lewis rat
Authors: Quintanar‐Stephano Andrés , Contreras‐Romo Martha Citlalli , Ibarra‐Pedroza Janeth Alejandra , Zambrano Cynthia , Organista‐Esparza Alejandro , Rodríguez‐Peralta Adriana , González‐Ruíz Claudia Rebeca , Kovacs Kalman , Berczi Istvan 
Published: June 2021

Susceptible Lewis rats to acute experimental autoimmune encephalomyelitis (EAE) develop a single episode of paralysis, spontaneous recovery and remain refractory to EAE re‐immunizations. In non‐susceptible rats, EAE induces a strong corticosterone (CO) response protecting against the disease, whereas in the susceptible Lewis rats an inappropriate CO response induces an altered immune response. Here we study the critical time in which CO induce the refractivity to relapses in the EAE by using Lewis EAE‐adrenalectomized (ADX) animals operated at different times of EAE‐immunization. Animals were immunized with guinea pig brain homogenate in CFA (200 μL subcutaneously).ResultsAll animals developed an acute episode of EAE symptoms which started at 10–11 days post immunization (DPI), however differences in the clinical course were apparent in the several groups. The Control group developed a single full‐blown disease without relapse. ADX 15 days before immunization and ADX 3 DPI groups developed a lethal form of EAE. ADX 20 DPI group developed a single full‐blown of EAE without relapse. ADX 27 DPI group developed the acute full‐blown symptoms with a full‐blown relapse. ADX 30 DPI group developed a mild but prolonged relapse. The ADX 36 DPI group developed a single full‐blown disease without relapse. Results indicate that there is a critical period (between 20–27 DPI), in which CO suppress the relapses of the EAE.Supported by UAA‐PIBB11–5 and CONACYT‐62317. México.
Neuroprotective effects of EGCG on H2O2‐ and MPTP‐stressed PC12 cells
Authors: Snitsarev Vladislav , Andrej Anastasia , Rotella David P 
Published: June 2021

Parkinson's disease results from degeneration of dopaminergic neurons. EGCG (epigallocatechin gallate), the most abundant catechin in tea and a powerful antioxidant, has been shown to play a neuroprotective role in the presence of degenerative stress inducers. We hypothesized that EGCG would protect a dopaminergic PC12 cell line from H2O2 and MPTP (1‐methyl‐4‐ phenyl‐1,2,3,6‐tetrahydropyridine)‐induced stress. PC12 cells were plated at 104 cells/cm2 in 60 (35 mm) wells. 15 wells were treated with 10 nM EGCG, 15 with 100 nM EGCG, and 15 with 1000 nM EGCG. 5 wells of each EGCG concentration were used as controls with no additional treatments. 5 wells of each EGCG concentration were treated with 100 μM H2O2, 5 wells of each EGCG concentration were treated with 10 μM MPTP, and 5 PC12‐ containing wells were used as the control with no treatment of EGCG. 5 wells were treated with 10μM MPTP only and 5 wells were treated with 100 μM H2O2 only, for a total of 60 wells. The PC12 cells with respective treatments were incubated for 24 hours, and 4 random images of each well were taken, for a total of (60×4) 240 images.The above figures display PC12 cells treated with maximum concentrations of H2O2 (100 μM) and EGCG (1000 nM) for 24 hours. The results indicate that EGCG plays a neuroprotective role in the presence of oxidative stress stimuli. Similar results were obtained after treating the PC12 cells with MPTP.
Increased cortical CCL3 expression is associated with microglia activation and can be ablated by reducing circulating TGFβ1 during hepatic encephalopathy
Authors: McMillin Matthew , DeMorrow Sharon 
Published: June 2021

Hepatic encephalopathy (HE) is a neuroinflammatory complication associated with liver failure. Other neuroinflammatory disorders show involvement of chemokine ligand 3 (CCL3) and its receptor chemokine receptor 1 (CCR1). Transforming growth factor beta 1 (TGFβ1) modulates these proteins and is elevated in the serum during HE. Our aims were to examine CCL3 and CCR1 expression in the cortex and their interactions with TGFβ1 signaling during HE. Male C57Bl/6 mice were injected with azoxymethane (AOM) to induce HE. Neutralizing antibodies against TGFβ or clodronate liposomes were injected prior to AOM. Cognitive impairment was monitored and at coma brains and livers were dissected. CCR1, CCL3 and IBA1 expression was assessed by immunofluorescence and immunoblotting. Microglia activation occurred following AOM treatment and microglia depletion with clodronate liposomes delayed neurological decline in AOM mice. Cortical CCL3 protein, but not CCR1, was significantly elevated in AOM mice. Treatment with neutralizing antibodies against TGFβ delayed neurological decline and decreased CCL3 protein with no effects on CCR1. Our data suggest that microglia activation following AOM could be due to upregulation of CCL3 and that neutralizing antibodies against TGFβ are neuroprotective possibly through the suppression of CCL3 protein in AOM mice.Research support is provided from NIDDK via 7R01DK082435–03.