bluuebunny/crossref_metadata_2025_split huggingface dataset:
Adipose tissue p50α/p55α abundance may drive acute high fat diet induced insulin resistance
Authors: Clayton Zachary , Hetrick Byron , McCurdy Carrie 
Published: June 2021

Insulin resistance (IR), a metabolic feature of obesity, drives a number of diseases. Recently, the focus of obesity research has been adipocyte‐immune cell crosstalk and its influence on systemic insulin sensitivity (IS). Currently, obesity induced IR is thought to arise secondary to inflammation due to infiltration of adipose tissue (AT) with pro‐inflammatory immune cells. Obesity is a progressive disease, and metabolic alterations in AT following acute nutrient excess, prior to development of obesity, are not well understood. Phosphotidylinositol 3‐kinase (PI3K) has been proposed as the integration point between immune cell infiltration and adipocyte insulin action. Global knockdown of pik3r1 (gene encoding regulatory subunits (p85α/p55α/p50α) of PI3K) resulted in enhanced adipocyte and systemic IS, with parallel decrease in AT macrophage infiltration despite marked obesity. Here we demonstrate that acute (3d) high fat diet (HFD) significantly increases bodyweight due to a significant increase in food intake in wild type mice. Body composition analysis by NMR indicates a significant increase in percent body fat with a concomitant decrease in lean mass. Energy expenditure measured by indirect calorimetry was also significantly decreased after 3d HFD. Importantly, both systemic IS measured by oral glucose challenge and insulin‐stimulated adipocyte 2‐deoxyglucose uptake was significantly reduced. No difference was found in adipocyte chemokine or cytokine secretion. We propose that reduction in adipocyte IS and subsequent systemic IS, is due to an increase in AT p50α/p55α abundance and occurs prior to initiation of a pro‐inflammatory response. Identifying primary steps in development of IR may guide treatments for obesity related metabolic diseases.NIH DK095926 (CEM)
Proteomic Analysis of the Late Effects of Intrauterine Undernutrition on Adipose Tissue of Rats
Authors: Souza Adriana , Pedroso Amanda , Tashima Alexandre , Ribeiro Eliane 
Published: June 2021

Poor nutrition during early development may program adulthood diseases such as obesity. This study performed proteomic analysis of the retroperitoneal adipose tissue of adult male and female rats born from dams either fed ad libitum or restricted to 50% of ad lib intake during pregnancy. Tissue proteins were analyzed by 2‐D gel electrophoresis and Maldi‐Tof/Tof – MS. Mascot server was used to search Swiss‐Prot.Results: The restricted female progeny developed obesity and had 18 down‐regulated and 4 up‐regulated proteins. The restricted males did not develop obesity but still had 12 down‐regulated and 17 up‐regulated proteins. Although the majority of the differentially expressed proteins are related to metabolic processes and have catalytic activity , gender differences were evident. For example, cytoplasmic malate dehydrogenase was up‐regulated only in males while glyceraldehyde‐3‐phosphate dehydrogenase was up‐regulated only in females.ConclusionFemale but not male rats develop into obese adults when undernourished intrauterus. Important metabolic processes in adipose tissue are programmed by intrauterine undernutrition with different phenotypes between male and female adult rats. These may help explain the gender dimorphism in the obesogenic response.Funding: FAPESP, CNPq, Capes: Brazil
GLP1R Regualtion of Gap Junction Coupling in the Islet of Langerhans
Authors: Walter Rachelle , Farnsworth Nikki , Benninger Richard 
Published: June 2021

Pancreatic islets regulate glucose homeostasis through insulin secretion. β‐cells are connected by gap junctions, which regulate depolarization, calcium signaling and insulin secretion. In diabetes, pro‐inflammatory cytokines contribute to β‐cell mass depletion and dysfunction. Low levels decrease Gap Junction Coupling (GJC) in the islet, disrupting calcium signaling and insulin secretion dynamics. Cyclic adenosine monophosphate (cAMP), elevated by a Type 2 Diabetes treatment called Exendin‐4, has been shown to increase GJC. Increased cAMP acts via protein kinase A (PKA) and exchange protein activated by cAMP 2 (Epac2) to regulate GJC in other systems; however, this has not been shown in pancreatic islets. We hypothesized Exendin‐4 can overcome cytokine induced decreases in GJC through PKA and Epac2 dependent mechanisms.To test this, isolated mouse islets were cultured overnight with a cocktail of pro‐inflammatory cytokines, 10nM Exendin‐4 and activators and inhibitors of PKA and EPAC2. Changes in GJC were quantified with Fluorescent Recovery After Photo‐bleaching, and calcium signaling was measured with calcium imaging of Fluo‐4.Cytokine+Exendin‐4 increased GJC compared to the cytokine treatment alone. PKA inhibitor decreased the Exendin‐4 recovery and had higher GJC than the cytokine treatment. PKA activator also recovered cytokine induced GJC decreases. Epac2 had little effect on recovery.These results suggest that Exendin‐4 is acting through PKA to overcome cytokine induced decreases in GJC while EPAC2 has minimal effect. Future work is needed to determine how PKA increases GJC via Cx36 in this system. This work will be important for targeting decreased GJC in pre‐diabetic patients.
Stromal Loss of mRNA‐Binding Protein IMP1 promotes a Pro‐Tumorigenic Colon Tumor Microenvironment in vivo
Authors: Chatterji Priya , Hamilton Kathryn , Lundsmith Emma , Andreas Sarah , Rustgi Anil 
Published: June 2021

BackgroundInsulin‐like growth factor II mRNA‐binding protein 1 (IMP1) is an oncofetal protein that modulates mRNA stabilization, localization, and translation. Recent studies suggest that IMP1 may play a role in colonic mesenchymal stem cells during wound healing. We have demonstrated an oncogenic role for IMP1 in colorectal cancer (CRC) cells; however, it is not known how non‐epithelial IMP1 contributes to CRC in vivo.Methods: Using the azoxymethane/dextran sodium sulfate (AOM/DSS) model of colitis‐associated CRC, we evaluated tumorigenesis in mice with mesenchymal‐specific Imp1loss (Dermo1Cre;Imp1floxed mice;Imp1ΔMes). Furthermore, isolated mesenchyme from tumor‐bearing mice was evaluated for changes in growth factors and cytokines via antibody array.Results: We demonstrate that Imp1 ΔMesmice treated with AOM/DSS exhibit a significant increase in tumor number, size, and load compared to controls. Isolated mesenchmye from AOM/DSS‐ treated Imp1 ΔMes mice show, among other things, a significant increase in hepatocyte growth factor (HGF) expression, suggesting that IMP1 loss favors a pro‐tumorigenic environment, potentially via up‐regulation of the HGF pathway.ConclusionsOur results thus far indicate a novel, protective role for stromal IMP1 in colon tumorigenesis, likely via negative regulation of HGF signaling in the setting of colitis‐associated CRC. Future studies will determine the direct mechanisms for IMP1 regulation of the HGF pathway and may define a previously unknown, therapeutically tractable mechanism of stromal:epithelial crosstalk in CRC.Funding : NIH RO1 DK056645 (AKR), NIH K01 DK100485 (KEH) HHMI Medical Fellowship (ETL), NIH UO1 DK085551 (NIDDK/NIAID Intestinal Stem Cell Consortium)
Evaluating the Efficacy of a Video Dissection Guide Tailored to the Louisiana State University Health Sciences Center Gross Anatomy Curricula
Authors: Campeau Lisa , Mussell Jason 
Published: June 2021

In an effort to close the educational gap caused by low instructor to student ratios in gross anatomy labs, a new video dissector based on the required written text used at Louisiana State University (LSU), Grant's Dissector 15th edition, is being created by the gross anatomy faculty. As currently available video guides are more demonstrative and less instructional, our goal is easing the perfunctory dissection burden by describing dissection techniques and characteristic structures so critical dissection of historically difficult‐to‐identify structures can be carried out more quickly with less dissection damage. It is hypothesized that these LSU‐specific videos will be of greater aid to students in identifying structures than guides already in existence or than written text alone. To test this hypothesis, the class will be divided into 3 groups: written dissector only, traditional anatomy video specifically Acland's Video Atlas of Human Anatomy, and LSU video dissection guide. Accuracy of structure identification will be assessed by a simple questionnaire administered to all 3 groups. Either the students will identify a given structure or it will be located by the faculty if they cannot. Also, video group assignation will be compared to gross anatomy practical exam scores for each dissection block. It is hypothesized that assignment to the LSU video dissection guide will result in higher practical exam scores than either traditional anatomy video or written dissector alone.Source of Support: Dept of Cell Biology and Anatomy, Louisiana State University Health Sciences New Orleans, LA
ERCC1 and Thymidylate Synthase as Prognostic Biomarkers in Malignant Mesothelioma
Authors: Mezzapelle Rosanna , Miglio Umberto , Rena Ottavio , Mercalli Francesca , Veggiani Claudia , Paganotti Alessia , Buosi Roberta , Rinaldi Maurizio , Bianchi Marco , Boldorini Renzo 
Published: June 2021

The standard of care for malignant pleural mesothelioma (MPM) is a combination of platinum compounds and Pemetrexed. Still, the median survival of MPM patients is low (about 12 months). Excision Repair Cross Complementing Group 1 (ERCC1) acts by removing DNA adducts formed by platinum compounds, whereas Pemetrexed inhibits Thymidylate Synthase (TS), which is involved in DNA synthesis.Pleural biopsies were collected from 140 MPM patients (98 males and 42 females, mean age 68 years) before treatment initiation. Average disease specific survival was 13 months (range 1‐60). mRNA was extracted from formalin fixed paraffin blocks, reverse transcribed and assayed by qRT‐PCR. Immunohistochemistry with anti‐ERCC1 and anti‐TS mAbs was scored semi‐quantitatively.Of 140 patients, 102 were treated, the remaining were not because of bad performance status or advanced age. Patients with low ERCC1 protein expression survived longer (24.3 vs 12.5 months, P=.03). Likewise, patients with low levels of TS mRNA had better survival (P=.01); there was no advantage, however, in the treated sub‐group, suggesting that Pemetrexed treatment and low TS expression had no additive value.ERCC1 protein expression and TS gene expression both seem important prognostic markers; we suggest their evaluation in routine biopsies from MPM patients.
Association between DAPK1 Promoter Methylation and Cervical Cancer: a Systematic Review and a Meta‐analysis
Authors: Agodi Antonella , Barchitta Martina , Quattrocchi Annalisa , Maugeri Andrea , Vinciguerra Manlio 
Published: June 2021

DNA methylation, a major epigenetic mechanism, is involved in various biological processes including cancer. Particularly, promoter methylation can inactivate Death‐Associated Protein Kinase1 (DAPK1), a tumor suppressor gene which is often methylated during the carcinogenesis of cervical cancer. In the present systematic review and meta‐analysis, we evaluated the association between DAPK1 promoter methylation and cervical cancer risk. A literature search, using the PubMED database, was performed to select studies published up to July 2014. Subgroups analyses were performed. A total of 27 studies were included. All studies evaluated DAPK1 promoter methylation in squamous cell carcinoma and 12 studies also in adenocarcinoma. The “gold standard” method of promoter methylation evaluation was methylation specific PCR. Twenty studies, evaluating DAPK1 promoter methylation both in tumor and in healthy controls samples, were included in the meta‐analysis (1092 from cancer patients and 837 from controls). Meta‐analysis showed a significant association between DAPK1 promoter methylation and cervical cancer (pooled OR: 21.20; 95% CI: 11.14, 40.35) with the random effects model. The association was confirmed in specific subgroups. In conclusion DAPK1 promoter methylation might play an important role in the pathogenesis of cervical cancer. The promoter methylation of DAPK1 is a frequent event in cervical carcinogenesis and may have potential clinical application as a cancer biomarker.This work was conducted with the research support of Bench Srl, University of Catania.
Anti‐Proliferative Effects of Selected Dietary Polyphenols on Human Colon Carcinoma Cell Line, T84
Authors: Javed Fatima , Khan Nabihah , Detloff Sally Jo , Sarathy Jayashree , Ronkainen Niina 
Published: June 2021

Polyphenols are naturally occurring compounds in plants which are typically involved in defense against ultraviolet radiation or pathogens. Previous studies have shown that various polyphenols have anti‐proliferative properties in a variety of human cancers. The purpose of this study is to determine the effect of Quercetin (QC), Epigallocatechin‐3‐gallate (EGCG), Catechin and Resveratrol (RVL) on cell death and apoptosis in human colon carcinoma, T84, cells.Cell death and apoptosis was measured by using Annexin V: FITC Apoptosis Detection Kit I and a flow cytometry. Short term (1 hour) and long term (overnight; O/N) effects were observed. Cells were grown in 6 well plates, serum starved O/N and treated with different concentrations of polyphenols for varying time. Linear Sweep Voltammetry (LSV) was used to electrochemically characterize the antioxidant capacity and to quantify active RVL and QC (the polyphenols acting as reducing agents) for the time frame of the study.While EGCG and Catechin did not have an effect, RVL and QC showed an interesting dose and time‐dependent change in cell death and apoptosis. O/N exposure to 200 µM QC caused an increase in cell death and apoptosis (control: 5.5+ 1.0%; QC: 15.5+ 4.3%). However, 30 µM RVL increased cell death and apoptosis at 1 hr (control: 4.3+ 0.8; RVL: 13.9+ 3.2) and returned back to control levels with time. Based on LSV, QC has a higher antioxidant capacity than RVL. However, the antioxidant effect of RVL decreased over time. In summary, QC and RVL both have anti‐proliferative properties with QC having prolonged effects in human colon carcinoma cells. Thus, a combination of polyphenols might have a better effect in preventing colon cancer.
The Role of En­doplasmic Reticulum–Associated Degradation in the Regulation of a Potassium Channel Associated with Type II Bartter Syndrome
Authors: O'Donnell Brighid , Mackie Timothy , Brodsky Jeffrey 
Published: June 2021

Type II Bartter Syndrome is an autosomal recessive disease caused by a loss of function mutation in the human KCNJ1 gene. This gene encodes the Renal Outer Medullary Potassium channel (ROMK), the primary route for K+ efflux within the nephron. To maintain K+ homeostasis, renal epithelial cells must regulate the apical surface density of ROMK. The molecular defects in ROMK channel trafficking and regulation that cause Bartter Syndrome are poorly defined, but we propose that some of the disease‐causing mutations lead to protein misfolding. Because misfolded membrane proteins are subject to ER quality control, we asked if some mutations target ROMK for En­doplasmic Reticulum–Associated Degradation (ERAD) in a yeast model. The ability of wild‐type and Bartter ROMK mutants to rescue low K+ sensitivity was investigated. Yeast lacking Trk1 and Trk2 K+ channel proteins were transformed with mutant ROMK, serially diluted, and plated onto media of various K+ concentrations. Growth was scored via imaging, and relative ROMK expression levels were assayed by Western blot. Wild type ROMK and Bartter mutant stability was analyzed by cycloheximide chase analysis. To half of each culture, MG132 was added to inhibit the proteosome, and protein translation was halted with cyclo­heximide. Aliquots were collected at various time points, and relative levels of ROMK were assessed by Western blot. We found that yeast expressing ROMK Bartter mutants exhibit growth defects on low K+ media, and the mutant proteins are less stable compared to wild‐type ROMK. Degradation is proteasome‐mediated. Bartter Syndrome mutants increase targeting for ERAD when compared to wild‐type ROMK.NIH Grant 5T32DK071492‐09 and R01GM075061
Three‐dimensional Domain Swapping in the α Subunit of Tryptophan Synthase
Authors: Michalska Karolina , Bigelow Lance , Endres Michael , Joachimiak Andrzej 
Published: June 2021

Tryptophan synthase is an enzyme participating in the last two steps of tryptophan synthesis in plants, fungi and bacteria. It consists of two protein chains, α and β, encoded by trpA and trpB genes, that function as an αββα complex. Structural and functional features of tryptophan synthase have been extensively studied, explaining roles of individual residues in catalysis and allosteric regulation of the two active sites. TrpA has become a model system to investigate folding mechanisms of larger proteins. In 1969, Jackson and Yanofsky observed that typically monomeric TrpA forms a small population of dimers. The dimerization was postulated to occur through an exchange of structural elements of the monomeric chains – a phenomenon currently known as 3D domain swapping. Nevertheless, structural details of the dimeric form of TrpA protein remained unknown. We determined the crystal structure of TrpA that for the first time illustrates domain swapping in a TIM barrel fold. The exchanging fragment represents the N‐terminal section and includes H0‐S1‐H1‐S2 elements. The hinge region corresponds to loop L2 linking S2 with H2'. S2 and L2 carry the catalytic residues, Glu and Asp. As the S2 element is part of a swapped domain, architecture of the catalytic apparatus in α2 is regenerated from the two protein chains. The dimer interface corresponds to the α‐β interface in the tryptophan synthase complex, suggesting that the observed α2 dimer may not form a functional complex with the β subunit.This work was supported by the NIH grant GM094585 (MCSG) and the U.S. DOE, OBER, contract DE‐AC02‐06CH11357 (SBC).
Analysis of 5' and 3' snoRNA termini maturation in Saccharomyces cerevisiae
Authors: Matuszek Zaneta , Szczepaniak Sylwia , Kufel Joanna 
Published: June 2021

Small nucleolar RNAs (snoRNAs) are the best characterized non‐coding RNAs transcribed by RNA polymerase II. They are produced as precursors, whose extended 3' ends are trimmed exonucleolytically, whereas 5' ends either undergo combined endo‐ and exonucleolytic processing by Rnt1 and Rat1, respectively, or remain unchanged. In the latter case, the m7G cap becomes hypermethylated by Tgs1. [figure1]cRT‐PCR analyses suggest that processing of snoRNA 3' and 5' termini is tightly coupled. Inhibition of 5' end maturation, in rnt1Δ strain, cause the accumulation of snoRNA precursors that don't possess mature 3' ends but carry polyadenylated extensions. This defect is further increased when components of the cap‐binding complex (CBC) or methyltransferase Tgs1 are missing. Although we observe an accumulation of pre‐snoRNAs in rnt1Δ strain, there are also mature snoRNAs present. These results, obtained in Northern blot analysis, suggest the existence of an alternative pathway of 5' end processing, that most likely involves pre‐snoRNA cap removal by Dcp1/Dcp2 complex. Interestingly most pre‐snoRNAs that accumulates in dcp2Δ strain, are not cleaved by Rnt1, which was confirmed by 5´ RACE analysis.Obtained data strongly suggest the existence of a quality control mechanism that coordinates 5' and 3' end processing.Research project was financially supported by by “Generacja Przyszłości” Ministry of Science and Higher Education Funding Programme.
Changes in fMRI Hypothalamic Activity Via Intrinsic Oscillations After Intake of Regular Soda, Diet Soda or Water During a Meal
Authors: Bravo Stephen , Lowndes Joshua , Rippe James 
Published: June 2021

The aim of this study was to examine hypothalamic changes after intake of different beverages at normal levels of consumption.A total of 63 healthy, non‐diabetic, normotensive subjects with BMI 21‐29.9 kg/m2, between the ages of 20‐50 years took part in an fMRI scanning session. The protocol consisted of six runs of resting‐state‐fMRI. During the first two runs subjects were in the fasting state. Subsequently they ate a standardized meal for 30 minutes outside the scanner that included either a regular soda, a diet soda, or water, based on random allocation (n=21 for each group). After the meal, subjects were scanned four additional runs. For data analysis, standard image preprocessing was performed using FSL software. We used fractional amplitude of low‐frequency fluctuation (fALFF), a validated method that allows the detection of the regional intensity of spontaneous low fluctuations (0.01–0.08 Hz) in fMRI BOLD signal. Voxelwise fALFF maps were calculated for each participant in native space, and then transformed into the MNI152 standard brain space. Before statistical analyses, each individual fALFF map was Z‐transformed. A seed of 3 mm of hypothalamus based on a previous study was used to extract the Z values from these fALFF maps for group‐level parametric analyses. We modeled the data using ANOVA with group (regular soda, diet soda, water), and condition (fast, fed) as factors.There was no effect of metabolic condition (F=1.99, p=0.16) or group (F=0.77, p=0.46). The interaction group x condition was also not significant (F=0.57, p=0.56). These results suggest that the previously identified differences may not apply to the case of SSBs under conditions that are more representative of normal daily intake levels and patterns.
Frequency of Metabolic Syndrome in a Sample of Brazilian Institutionalized Elderly
Authors: PEDROSA LUCIA , Sales Marcia , Sena‐Evangelista Karine , Oliveira Larissa , Lemos Telma , Santos Ana , Lima Kenio 
Published: June 2021

Objectivemeasure the frequency of MS in institutionalized elderly in Natal, Northeastern Brazil.Methodsa cross‐sectional study was conducted in elderly persons aged 蠅 60 years, from 12 nursing home care facilities. The frequency of MS was estimated according to the National Cholesterol Education Program ‐ Adult Treatment Panel III. Participants were clinically assessed for fasting glycemia, lipid profile, body mass index, waist circumference, physical activity level, smoking and food consumption.ResultsThe study enrolled 203 elderly with mean age 81.97 (SD=9.11) years, 75% women and 72% residing in public nursing home care. The frequency of MS was 29.6%, low HDL cholesterol levels being the most frequent component (28%). Physical inactivity, overweight and altered waist circumference was observed in 73%, 33%, and 40% of the total sample, respectively. Habitual diet showed mean energy intake of 1394 kcal (SD=356) with high percentages of adequate ingestion of carbohydrate (74%) and saturated fat (97%). The associated factors of MS were age between 80 and 90 years (PR=7.64; 95% CI 1.86‐31.37) and 蠅 90 years (PR=6.22; 95% CI 1.40‐27.59); being female (PR=0.42; 95%CI 0.19‐0.94) and diabetes (PR= 14.01; 95% CI 6.29‐31).Conclusionsthe study population exhibited moderate prevalence of SM involving associated factors that should be considered in the care of the health of elderly individuals.