bluuebunny/crossref_metadata_2025_split huggingface dataset:
Using Human Stem Cells to Analyze the Effects of the Immune System on Chronic Kidney Disease
Authors: D'Agostino Victoria Anne , Dunkley Truman , Toth Charles 
Published: April 2020

Recent studies have shown that there exists a correlation between Chronic Kidney Disease (CKD) and macrophage‐induced inflammation. Utilizing human induced pluripotent stem cells (hiPSC), the inflammatory process in kidney disease could be recapitulated and the mechanisms behind it better understood. Kidney organoids and macrophages were grown from two hiPSC cell lines, BJFF.6 and WTC11, following previously described procedures. An inflammatory response was induced by the addition the pro‐inflammatory mediator Poly (I:C) and Lipopolysaccharide (LPS). The role of the microbiome in kidney disease was examined by the introduction of the tryptophan metabolite, indole‐3‐sulfate (I3S). The difference in gene expression in the kidney and macrophage co‐cultures for two inflammatory markers, TNFα and IL‐6, were analyzed using qPCR. ELISA analysis was also performed, which showed that there was an overall increase in inflammation in the co‐cultured kidney organoids, which was reduced in the presence of 100 uM I3S. This demonstrates that microbiome metabolites can influence an inflammatory response. Examining the role of the microbiome in an inflammatory disease model can be used to further study kidney inflammation and detect potential targets for treatment of CKD.Support or Funding InformationProvidence College Undergraduate Research Grant, and a gift from Dr. John Mullen
Resveratrol Attenuates Doxorubicin Cytotoxicity and Mitochondrial Dysfunction and Human Proximal Tubular Epithelial Cells
Authors: Valentovic Monica A. , Brown Kathleen C. , Dial Mason , McGuffey Rachel 
Published: April 2020

Doxorubicin (DOX, Adriamycin), is a cancer chemotherapy employed in the treatment regimen for breast, Hodgkin and Non‐Hodgkin lymphomas, small cell lung cancer and acute lymphoid leukemia. The U.S. FDA issued a Black Box warning for Cardiomyopathy in patients treated with doxorubicin. Other adverse effects include renal impairment. The cellular mechanisms for doxorubicin mediated renal cytotoxicity are not known. Resveratrol (RES) is a phytoalexin found in many fruits including grapes, blueberries, cranberries, dark chocolate and nuts. Antioxidant and anticancer properties have been associated with RES in cell culture and in human clinical studies in cancer patients and in vivo rodent models. Our studies tested the hypothesis that RES protection for DOX renal cytotoxicity, was mediated by maintaining mitochondrial function. All studies were conducted in human noncancerous renal proximal tubular epithelial cells (HK‐2). HK‐2 cells were plated and equilibrated for 48 h. Cells were next pre‐incubated for 1 h with 0 (DMSO) or 5 or 7.5 uM RES followed by a 24 h co‐incubation with 0–5 uM DOX. All results were obtained from 3 independent experiments. Western analysis for protein carbonylation and 4‐hydroxynooneal (4‐HNE) were used as an indicator of oxidative stress. Mitochondrial function was assessed using a Seahorse analyzer system. Prior to examining mitochondrial function, cell number and substrate concentrations were optimized based on the manufacturer’s recommendations. Mitochondrial Stress Test was conducted following the treatments described above and each well was normalized by protein. Viability was assessed using MTT leakage and tyrpan blue exclusion cell counts. RES did not alter cell viability as indicated by comparable MTT values between DMSO and RES groups (p>0.05). DOX was cytotoxic to HK‐2 cells within 24 h. Pretreatment with RES provided protection from DOX to HK‐2 cells. Basal mitochondrial respiration assessed as oxygen consumption rate (OCR) was diminished by 4 uM DOX. Mitophagy mediated by DOX and the effect of RES compared protein expression of LC3BI, LC3BII and the ratio of LC3BII/I. In summary, RES attenuated DOX renal cytotoxicity by maintaining mitochondrial function. Further studies are needed to probe changes in specific mitochondrial proteins mediated by DOX.Support or Funding Information(Supported by NIH Grant P20GM103434 to the West Virginia IDeA Network for Biomedical Research Excellence; M.D. and R.M. supported by WV NASA Undergraduate Research Fellowship: M.D. Supported by MUSOM Summer Research Fellowship).
Faculty Perceptions of the Implementation of Flipped Teaching in Undergraduate STEM Courses Across Two Institutions
Authors: Givens Adriana , Bracey Georgia , Fickas Julie , Bartels Lynn , Locke Sharon , Gopalan Chaya 
Published: April 2020

The flipped classroom is an active learning and teaching environment that has been shown to improve student engagement and thus student performance. In order to expand the use of flipped teaching (FT) in STEM classrooms, we recruited faculty members from two institutions, six faculty from a local community college and an additional six from a regional state university. We were particularly interested in how faculty perceived the implementation of FT, barriers in the implementation of FT, and the differences in FT implementation at a four‐year university and a two‐year community college. All 12 faculty members participated in six faculty development FT workshops, that were spread over an entire semester. While undergoing training, the participants were expected to be preparing for their implementation of FT during the following semester. Pre‐implementation survey data from both institutions showed that 6 out of 12 faculty had a strongly favorable attitude toward FT and 4 out of 12 strongly agreed that they possessed the knowledge to use FT. When implementation of FT was considered, 4 out of 6 community college faculty and 2 out of 6 university faculty strongly agreed that they could successfully implement FT. One common barrier contributing to the resistance in the implementation of FT as identified by the participants at both institutions in the pre‐implementation survey was the lack of time to implement. Additional barriers identified specifically by the university faculty included lacking the knowledge to implement FT as well as anticipating potential resistance from students. Post‐implementation survey data showed that university faculty attitudes toward FT increased from 3 out of 6 to 5 out of 6 while community college faculty’s attitudes remained at 3 out of 6. There was an increase in knowledge of using FT from 2 out of 6 to 6 out of 6 by university faculty and from 2 out of 6 to 5 out of 6 by community college faculty. Additionally, faculty perceptions of successful implementation increased from 2 out of 6 to 5 out of 6 by university faculty and remained unchanged at 4 out of 6 among community college faculty. Multiple barriers were identified through post‐implementation survey by faculty from both institutions which included lacking the time to implement, limited knowledge related to implementation, grading, and class sizes. An additional barrier identified specifically by the university faculty was anticipating potential resistance from students. Addressing these barriers is critical in allowing the use of active learning methodologies such as FT.Support or Funding InformationNational Science Foundation
CRF Release in the Nucleus Accumbens Promotes Arousal in Anticipation of Cocaine Availability: Implications for Stress‐Induced Drug‐Seeking
Authors: Leonard Michael , Covington Herb , Miczek Klaus 
Published: April 2020

Dopamine signaling within the nucleus accumbens core (NAcC) is critical to the generation of motivated behaviors in both appetitive and aversive contexts. Within the accumbens, various neuropeptides influence motivational states through the modulation of dopamine release dynamics. Here, microdialysis studies show that the stress peptide corticotropin releasing factor (CRF) is released in the NAcC when rats are exposed to environmental contexts that predict either an aversive outcome (i.e. social defeat) or drug‐reward opportunity (i.e. cocaine availability), and moreover, intra‐NAcC infusion of exogenous CRF elicits a robust increase in extracellular dopamine. The present studies further interrogate accumbens‐specific CRF actions as a candidate mechanism for maladaptive arousal evoked by drug‐predictive stimuli – a hallmark feature of cocaine dependence. In order to assess the role of NAcC‐CRF on instrumental responding directed towards cocaine procurement, male Long‐Evans rats were trained to self‐administer cocaine under a heterogeneous chained schedule of reinforcement (FIFR) in order to dissociate anticipatory (‘drug‐seeking’) from consummatory (‘drug‐taking’) behavior. Completion of a fixed‐interval (10 min) was followed by a period (10 min) of continuously reinforced responding (0.32mg/kg cocaine; FR1) on another lever. Microdialysis conducted during this procedure revealed that extracellular dopamine levels within the NAcC gradually ‘ramp up’ across the fixed‐interval in parallel with lever pressing activity, as drug availability approaches. Similarly, dialysate levels of CRF were specifically and progressively elevated during the fixed‐interval component preceding drug‐access, but were unaffected by drug‐taking. Infusion of CRF into the NAc produced a rapid elevated tonic dopamine levels that was accompanied by enhanced responding during the fixed‐interval component of the procedure, but notably did not affect subsequent cocaine intake. Conversely, pharmacological blockade of CRF‐R2, dampened extracellular dopamine levels in the NAc and suppressed lever pressing behavior during the fixed‐interval. Taken together, these data suggest that in response to appetitive or aversive stimuli, CRF may be a critical mediator of motivated processes that are classically associated with dopamine transmission. To this end, stress‐evoked CRF release in the NAc may represent a candidate mechanism by which stress triggers aberrant cocaine‐seeking.Support or Funding InformationNIDA R01‐DA031734 to KAM
Dicer as a Repressor of Antiviral Responses in Mouse Embryonic Stem Cells
Authors: Gurung Chandan , Sapkota Krishna , Huang Faqing , Guo Yan-Lin 
Published: April 2020

Recent studies have demonstrated that pluripotent stem cells (PSCs), including human and mouse embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs) have underdeveloped antiviral innate immunity. Specifically, they fail to express IFN in response to viral stimuli. This finding has led to the hypothesis that the lack of IFN response is an intrinsic property of PSCs. It has also inspired the investigation of the underlying molecular mechanisms. In an effort to conduct a holistic analysis of antiviral responses in ESCs, we have developed an assay that can simultaneously measure RNA‐mediated gene expression and RNA‐induced antiviral responses. In this system, GFP is expressed from functionalized synthetic GFP‐mRNA and is used to measure the cellular translation activity, whereas dsRNA and 5’‐triphosphate‐ssRNA act as viral RNA analogs. Transfection of ESCs with these synthetic RNAs can potently activate all common antiviral signaling pathways, including RNA interference (RNAi), dsRNA activated protein kinase (PKR), OAS‐RNase L, and IFN responses. Our data revealed that RNAi activity was detected in ESCs, but not in Dicer knockout ESCs (D‐/‐ESCs) as expected since Dicer is a key enzyme in miRNA and siRNA processing. The antiviral responses mediated by PKR, OAS‐RNase L, and IFN were either not detected or detected at low levels in ESCs, but they increased dramatically in D‐/‐ESCs, as indicated by increased activity of PKR (measured by low level GFP expression and PKR phosphorylation) and RNase L (determined by RNA degradation). Furthermore, the components of the IFN signaling pathway are expressed at higher basal levels in D‐/‐ESCs than in normal ESCs and are further up‐regulated by RNA transfection. The most notable effect of RNA transfection into D‐/‐ESCs is the induction of cell death and decreased cell proliferation, which is at least partly attributed to PKR activation and increased expression of the cell cycle inhibitors p21 and p19. Our findings suggest that ESCs are exquisitely sensitive to cellular damages caused by antiviral response and that Dicer acts as a repressor of antiviral responses in ESCs under normal conditions.Support or Funding InformationThis work was in part supported by the National Institute of General Medical Sciences (R15GM128196‐01). We thank Mississippi‐IDeA Network of Biomedical Research Excellence for the use of the imaging facility (funded by the National Institute of General Medical Sciences P20 GM103476‐11).
Enhanced Mitochondrial Turnover in Aged Human Right Atrial Tissue
Authors: Oliveira Ashley N. , Karmanova Lioubov , Murugavel Shamini , Yanagawa Bobby , Hood David A. 
Published: April 2020

Age is one of the greatest risk factors for cardiovascular disease, which is the leading cause of death in the developed world. Therefore, the maintenance of cardiac health throughout the lifespan is of the upmost importance. Aging is associated with a plethora of biochemical and metabolic alterations, many of which converge on the mitochondrion. As a highly oxidative tissue, mitochondria supply roughly 95% of all ATP required in the myocardium. This reliance on mitochondria underscores the importance of maintaining the quality of these organelles, especially with age, however little is known about how age effects quality control mechanisms in the human heart. We hypothesized that the removal of mitochondria through mitochondrial specific autophagy, or mitophagy, would be impaired in aged human cardiac tissue. To evaluate mitochondrial quality control, samples of right atria were collected from young (≤50 years) and aged (≥70 years) patients undergoing coronary artery bypass surgery (CABG). Tissue was collected prior to and immediately following surgery, providing a useful model of ischemia‐reperfusion injury. Patients were matched for hyperention and dyslipidemia, as well as prescribed medications such as statins, and ACE inhibitors. Exclusion criteria included smoking, a prior stroke or myocardial infarction, or other comorbidities (e.g. diabetes, cancer). Mitochondrial content as indicated by various markers such as COX IV, VDAC, and UQCRC2 was reduced in the aged samples, however a modest 1.2‐fold increase in Citrate Synthase was observed, suggesting age‐associated compositional changes. The upstream marker of autophagy Beclin‐1 and the transcriptional regulators TFEB and TFE3 did not change with age, suggesting that aging does not affect the drive for autophagy. Despite this, constituents of the autophagosome p62 and LC3‐II were reduced by 20% and 30%, respectively, in the aged samples, indicative of increased autophagy flux. The mitophagy marker Parkin displayed a trend to decrease with age, suggesting a decline in the signaling for mitophagy. The lysosomal marker Cathepsin D was unaffected by age in cardiac muscle, but the lysosomal calcium channel Mucolipin‐1 increased in aged atrial muscle by 2‐fold. Analysis of post‐CABG samples indicated marked elevations in HSP70 and Caspase‐3 protein compared to pre‐CABG values. This was only evident in the atria of aged individuals. Our data suggest that increased mitophagy flux could account for the reductions in mitochondrial markers in aged atria, and that CABG can induce a stress response leading to apoptosis signaling that is more prominent with age.Support or Funding InformationSupported by NSERC Canada.
Characterizing the Action of Arginine Vasopressin at Hypoglossal Motoneurons In Neonatal Mice
Authors: Bolte Kristen , Revill Ann L. 
Published: April 2020

Vasopressin, traditionally considered a hormone, also acts as an excitatory neurotransmitter. The effects of vasopressin have been characterized in brainstem motoneurons, including hypoglossal motoneurons (XII MNs). The XII motor nucleus receives inspiratory rhythm from the preBötzinger Complex where it is relayed to the muscles of the tongue via the XII nerve. With some variation across postnatal maturation, the vasopressin V1a receptor gene is expressed from birth to adulthood in the XII motor nucleus, and is enriched in the XII nucleus in adults compared to other cranial motoneuron populations. Previous research demonstrated that vasopressin, acting via V1a receptors on preBötzinger Complex, can stimulate breathing in adult rats by increasing breathing frequency as well as diaphragm and genioglossus (primary tongue protruder) muscle activity. Whether vasopressin can also act directly at XII MNs to potentiate inspiratory bursting has not been well characterized. Thus, the aim of this research was to characterize vasopressin effects on XII MN inspiratory bursting behavior. We generated rhythmic, transverse medullary slices (600μM) from CD1 mice of either sex (postnatal days 0–5), and recorded inspiratory activity from XII nerve rootlets using a suction electrode. We then bath applied arginine vasopressin (0.01μM and 0.1μM, AVP, V1a receptor agonist), or locally injected AVP (0.1μM and 1.0μM). In doing so, we were able to assess the effects of arginine vasopressin on XII inspiratory burst amplitude. Bath applied AVP increased inspiratory burst amplitude by 43.4%±12.5% (n=9) when applied at 0.01μM, and 56.4%±18.1% at 0.1μM. When locally applied (n=7), AVP at 0.1μM increased inspiratory burst amplitude 38.8%±6.2%, and AVP at 1.0μM caused a 26.5%±6.3% increase in inspiratory burst amplitude with a notable increase in tonic activity. In a separate set of experiments (n=8) to test whether AVP acts via V1a receptors, local AVP increased inspiratory burst amplitude by 40.2%±9.3%. Local pre‐application of the V1a receptor antagonist ((d(CH2)51Tyr(Me)2Arg8) Vasopressin, 500nM) for 60 s attenuated the response to AVP (6.7%±14.7% decrease in inspiratory burst amplitude), but the AVP response was recovered after 15 minutes with an increase in XII inspiratory burst amplitude of 22.3%±4.1%. Overall, our results support that AVP can increase XII inspiratory burst amplitude via V1a receptor activation. Further research will need to determine whether AVP plays a role in maintaining airway patency in vivo.Support or Funding InformationKenneth A. Suarez Research Fellowship, Midwestern University
Proteome and MicroRNAs in Human Milk: Term‐Dependent Changes in Exosome Composition
Authors: Vahkal Brett , Ferretti Emanuela , Altosaar Illimar , Beaulieu Jean-François 
Published: April 2020

Infants born prematurely are a vulnerable population. Human milk is considered the best source of nutrition for infants as it contains not only nutrients, but also a variety of bioactive components to help infants adjust to life outside the womb. Among these components are exosomes, which are lipid bilayer vesicles that contain messenger RNA (mRNA), microRNA (miR) and proteins. Human milk exosomes contain various immunomodulatory components that may provide protection and promote microbial tolerance in the GI tract of newborns. However, mothers birth infants at very different times. Could preterm birth signal mother’s mammary gland to manufacture magical milk? We propose that term and preterm human milk convey different exosome constituents to the infant gut. We hypothesized that term and preterm human milk have distinct protein and miR content, and the exosomal isolates have unique immunomodulatory effects on intestinal cells. To investigate this, exosomes were isolated from human milk after term and preterm births. The exosomal miRs were characterized using RNA sequencing. Exosomal proteome enabled determination of abundant protein classes, narrowing of protein profile and discovery of essential proteins in preterm versus term milk. To investigate downstream pathways of either exosomes or specific exosomal‐contained proteins, we used the Caco 2/15 intestinal cell line to measure levels of immunomodulatory proteins following treatments with milk derived exosomes or proteins. These results provide novel insights for adaptations that exist in human term and preterm milk on both the proteome and expressional level. Ultimately, this research can guide the development for therapeutics in infant gastro‐intestinal diseases, such as necrotizing enterocolitis, and elucidate unique proteins that are released into human milk for downstream action(s) in the developing infant. Therapeutic benefits of preterm milk constituents are being explored.Support or Funding InformationCanadian Institutes of Health Research
Mechanism of Action of Moringa oleifera (Moringa) in the Treatment of Diabetes Mellitus
Authors: Obih John-Clifford Agbasi , Obih Patience Ogechi , Arome Okai Samuel 
Published: April 2020

Diabetes mellitus is a metabolic disease characterized by impairment of carbohydrate, protein and fat metabolism which results from defect in the secretion of insulin, insulin utilization or both. According to CDC, 30.3 million people have diabetes in the USA out of which 23.1 million people have been diagnosed and 7.2 million people have not yet been diagnosed. Approximately 84 million American adults have prediabetes. The prevalence of diabetes is increasing both in developed and developing countries. Standard drugs are available in the market for the treatment of diabetes. However, many people with diabetes are turning to medicinal herbs for treatment. The assumption of those who use medicinal herbs is that the herbs are edible and do not carry attendant side effects as the drugs in the market. Moringa oleifera is among the medicinal herbs used locally for diabetes. It is widely cultivated in India. Research has shown that Moringa oleifera is an edible plant that has multiple benefits in diabetes. In many parts of Africa, moringa is widely used for self‐medication in diabetes and hypertension. Its mechanism of action as an antidiabetic has not been well elucidated. This study was undertaken to examine if moringa affects carbohydrate metabolism like acarbose that reduces hyperglycemia by inhibiting alpha‐glucosidase. The alpha‐glucosidase inhibition study of moringa aqueous leaf extract was performed in vitro in a 96 well‐plate using Saccharomyces cerevisiae as the source of alpha‐glucosidase. The assay was analyzed in a multiskan spectrophotometer and the absorbance was read at 400 nm. A parallel test was run with acarbose. Acarbose, gave a dose‐dependent inhibition of alpha‐glucosidase and moringa showed also activity of a similar mechanism, but not as potent as acarbose. It has been reported that herbal medicines have multiple mechanisms of action. More studies need to be carried out to identify other possible mechanisms of action of Moringa oleifera as an antidiabetic.Support or Funding InformationCOE