bluuebunny/crossref_metadata_2025_split huggingface dataset:
The effects of high fat diet‐induced obesity and exercise training on hematopoietic stem and progenitor cells following sub‐lethal radiation
Authors: Emmons Russell , Hernández‐Saavedra Diego , Parks Zachary S , Chen Hong , De Lisio Michael 
Published: November 2023

Of the 14 million cancer survivors in the United States, approximately 2/3 have received radiation therapy. Hematopoietic stem and progenitor cells (HSPCs) are among the most sensitive cells to radiation damage resulting in anemia and immunosuppression acutely following radiation exposure, and decreased hematopoiesis in the long‐term. Increased adiposity and physical inactivity are common among cancer survivors and both have negative effects on hematopoiesis. However, the impacts of obesity and exercise on the late effects of radiation exposure on HSPCs are unknown. The purpose of the present investigation was to evaluate the impact of obesity and exercise on HSPCs following sub‐lethal radiation exposure. A total of 40 CBA mice were divided into 4 groups (CON+SED n=10, CON+EX n=10, HF+SED n=10, HF+EX n=10). Mice were put onto a control (CON) or high fat diet (HFD) to induce obesity for 4 weeks before being placed into a progressive treadmill exercise (EX) program for 9 weeks or remained sedentary (SED). Four weeks into the training program, all mice were exposed to a 3 Gy dose of gamma radiation and completed their respective exercise and diet programs. At the conclusion of the 9 week intervention, bone marrow cells were collected and lineage negative cells were separated and analyzed for total HSPCs (LSK, SCA+cKit+), and HSPC subpopulations: long‐term (LT‐HSC, LSK CD150+CD48−), short‐term hematopoietic stem cell (ST‐HSC, LSK CD150+, CD48+), and multi‐potent progenitors (MPP, LSK CD150−,48+). HFD‐induced obesity reduced bone marrow cellularity compared to control mice (p<0.01). EX attenuated these changes to bone marrow cellularity (p<0.01). A decrease in the total number of LSK resulted from consuming HFD (p<0.01) that was attenuated with EX (p<0.01). Similar HFD‐induced decreases were seen in LT‐HSC (p<0.01), ST‐HSC (p<0.01), and MPPs (p<0.01) which were attenuated with EX (LT‐HSC, p<0.01, ST‐HSCs, p<0.05) and strongly trended in MPPs (p=0.067).ConclusionHere we show that HFD‐induce obesity decreases bone marrow cellularity and HSPC content following radiation exposure which is rescued by EX. As such, exercise training may serve as a promising therapy to preserve HSPCs following radiation therapy.Support or Funding InformationFunded by the UIUC Research Board Grant, ACSM Student Research Grant, and ACSM Foundation Grant
CL‐K1, a Novel Lectin Pathway Component is a Leptomeningeal Protein
Authors: González‐Losada Cristóbal , Castillo‐González William , Jensenius Jens Christian , Zerr Inga , Schmitz Matthias , Dorta‐Contreras Alberto Juan 
Published: November 2023

The universal validity of the molecular diffusion/cerebrospinal fluid (CSF) flow model is valid for the brain‐derived molecules, is the base for a systematic approach to determine the source of a CSF protein either derived from blood, from brain or from leptomeninges.ObjectiveTo determine if CL‐K1, a new component of the lectin pathway, should be consider a leptomeningeal protein.Methods36 serum and CSF paired samples were obtained from control persons without neuroinflammation process and without oligoclonal IgG bands. CL‐K1 and albumin were quantified by ELISA and by an immunoturbidimetric analysis. Q albumin= CSF albumin/serum albumin and QCL‐K1= CSF CL‐K1/serum CL‐K1 was calculated.ResultsSerum and CSF CL‐K1 follow a normal distribution according to Kolmogorov‐Smirnov test. Mean values of CL‐K1 in serum was higher than its mean content in CSF. CSF CL‐K1 concentration increased with the increase of Q albumin. Cl‐K1 values in serum are higher than in CSF. This fact notes that CL‐K1 is not a brain‐derived protein. In Figure 1 there is the relationship between CL‐K1 vs. Q albumin. Note that the regression line has a positive slope. It means that CL‐K1 increases with increment of Q albumin. It is a general rule that follows the proteins that was located in the leptomeninges.ConclusionsThese results means that the function is not solely diffusion dependent and allowed the finding that CL‐KI in CSF derives from the leptomeninges as well as mannan‐binding lectin.
A Novel Lipidomic Approach to Understand Human Diabetic Heart Disease
Authors: Pinz Ilka , Robich Michael P. , Ryzhov Sergey , Sawyer Douglas B. , Vary Calvin 
Published: November 2023

Diabetes mellitus is an important risk factor for acute myocardial infarction and chronic ischemic heart disease. We have demonstrated in an animal model that high fat feeding causes lipotoxic changes in the heart that includes the accumulation of sphingolipids, lipids that are enriched in specialized membrane microdomains called caveolae. Accumulation of sphingolipids causes the loss of caveolin proteins (CAV) from the plasma membrane, as well as the loss of receptor tyrosine kinase signaling, such as insulin and other growth and survival signaling. We have now expanded our work from animal models to informed and consented human diabetic patients who are scheduled to undergo coronary artery bypass grafting (CABG) surgery at Maine Medical Center. The protocol and use of human tissue samples for this study has been approved by the Institutional Review Board of Maine Medical Center. We tested whether disrupted membrane lipid homeostasis is associated with the loss of survival signaling in the diabetic heart, and thus may contribute to the increased risk of cardiovascular complications in diabetics. To test this idea, we used an unbiased mass spectrometric lipidomics approach to determine the lipid profiles in right atrial appendix (RAA) tissue and blood plasma, and determined the expression of CAV1, CAV3, IRS‐1, and ERBB2, 3 and 4 receptors in RAA by Western blotting. We found different lipidomic signatures in human diabetic vs. non‐diabetic blood plasma and right atrial appendage tissue demonstrated by different principle components in the global lipid comparison. A more detailed analysis showed that diabetic subjects have increased plasma and tissue levels of sphingolipids, which dependent on the specific sphingolipid species, are between 2 and 15 times higher compared to non‐diabetics (n = 5 in both groups). The expression of caveolin proteins and total ERBB receptors varied throughout our patient population (n = 8), however increased lipid levels correlated with lower RTK expression and lower caveolin protein expression. Flow cytometric analysis of isolated atrial endothelial cells showed reduced cell surface ERBB2 expression in diabetics compared with non‐diabetics.These data support the hypothesis that dysregulated lipid homeostasis in cardiac cell membranes of diabetic patients may be contributing to disrupted RTK signaling. To our knowledge this is the first study to characterize these changes in this patient population.
ob/ob Mice Fail to Lose Weight on an Alternate‐Day Fasting Diet, Yet Improve Glucose Control
Authors: Bingaman Mark Jake , Hult Elissa M , Swoap Steven John 
Published: November 2023

Alternate day fasting (ADF) has proven effective for weight loss in humans and rodents on a high‐fat diet. Mice with a homozygous mutation in the ob gene are obese, diabetic, and exhibit reduced body temperature (Tb) with variable torpor bouts in the fed state. We tested the hypotheses that an ADF protocol in genetically obese mice would result in (i) body weight (BW) loss, (ii) improved glucose control, and (iii) induction of torpor on fasted days. We evaluated the physiological effects of ADF in control and ob/ob mice for caloric intake, BW, Tb, and circulating glucose and insulin. Adult female ob/ob mice were implanted with temperature telemeters and randomly assigned to either 1) ad lib food availability (n=6) or 2) an ADF protocol, where the mice were fasted every other day, with ad lib feeding between fasts (n=11). Over 18 days of ADF, ob/ob mice consumed 258±11 kcal, approximately two‐thirds of that consumed by ob/ob mice on an ad lib diet (410±21 kcal). Despite this, ob/ob mice on the ADF diet lost only 0.7±0.2g of BW compared to the 3.9 ± 1.3g weight gain by ad lib ob/ob mice. ADF caused long and deep bouts of torpor in ob/ob mice such that the average and minimum 48‐hour Tb was 34.0±0.2 and 26.1±0.2°C, respectively, as compared to ad lib ob/ob mice (35.2±0.3; 31.1±0.9°C, respectively). Circulating glucose fell in the ADF mice, but only on fasting days (ad lib 307±57, ADF‐Fed 271±8, ADF‐Fast 132±17 mg/dl). The HOMA index, which evaluates insulin sensitivity using circulating glucose and insulin, improved significantly in the ob/ob ADF mice, but only on fasting days (ad lib 913±349, ADF‐Fed 1256±393, ADF‐Fast 288±146). These data demonstrate that weight loss is diminished, likely due to the deep bouts of torpor on fasted days, even though ADF significantly reduces food intake in ob/ob mice. Further, circulating glucose and the HOMA index fell in the ob/ob ADF mice on fasting days, suggesting improved glucose control in the face of minimal weight loss.Support or Funding InformationSupported by 1R15HL120072‐01A1 to SJS
Effects of Sleep Duration on Insulin Resistance – A Longitudinal Study in Healthy, Lean, College‐Age, Males
Authors: McGregor Kathryn , Cialkowski Kathryn , Townsend Dana K 
Published: November 2023

IntroductionThere are sparse interventional, within subject, clinical sleep studies. College‐age adults can experience substantial changes in sleep duration but have received little attention relative to potential concomitant changes in in fasting and dynamic measures of glucose metabolism.PurposeTo test the hypothesis that in healthy, lean males in a college‐age group (18–26 yrs.), that sleep attenuation measured in the home environment for 1 week, at a level approximate to average college‐age sleep deprivation (1.5 hrs. < 7–7.5 hrs./night), will significantly increase insulin resistance while augmentation for 1 week, to recommended average levels for this age group (8 hrs.), will significantly decrease insulin resistance.Methods17 healthy, lean, male subjects (age 21.1 ± 2.0 yr., BMI 22.4 ± 2.2) slept according to their normal schedule in the home environment (7hrs 22min ± 33min) for one week, slept 1.5 hours below their baseline (6hrs 7min ± 35min) per night on week two, and with sleep duration augmented to optimal sleep (7hrs 56min ± 28min), on week three. Sleep was measured by wrist actigraphy augmented with individual sleep logs. Following each week, fasted subjects underwent a two‐hour oral glucose tolerance test. HOMA‐IR, a measure of hepatic insulin resistance, and the Matsuda Index, a measure of whole body insulin sensitivity, were calculated each week. The average differences in values of the HOMA and Matsuda between baseline, short sleep, and augmented sleep were analyzed using one way repeated measures ANOVA.ResultsThere was no statistically significant effect of sleep augmentation or deprivation between baseline, short sleep, and long sleep on HOMA‐IR (p‐value=.06) and Matsuda Index (p‐value=.30) values.ConclusionThis study shows sleep reduction and augmentation, at moderate levels that approximate normal college‐age sleep, may not have significant short‐term impacts on insulin sensitivity in lean males.Support or Funding Information2015, 2016 Aldeen Award‐‐Wheaton College, office of the Provost 2015, 2016 Summer Science Research Student in Residence support‐‐Wheaton College, Alumni Association
Gα12 is required for renal cystogenesis through ADAM10 dependent junction dysfunction pathway
Authors: Xu Frank , Ho Li‐Lun , Kong Tianqing , Lu Tzongshi 
Published: November 2023

BackgroundAutosomal dominant polycystic kidney disease (ADPKD) is one of the most common life‐threatening genetic diseases and is characterized by early formation and gradual enlargement of multiple kidney cysts, which eventually results in end‐stage renal disease (ESRD). At ADPKD, cell‐cell junctions and polarity between epithelial cells are disrupted. Cadherins constitute a major protein family in polycystin‐1 (PC1) complexes in renal epithelial cells. And, E‐cadherin plays an important role to maintain the integrity of cell‐cell contacts and polarity. Our previous data showed that PC1‐regulated protein, Gα12, deletion protected kidneys from development of kidney cysts induced by Pkd1 inactivation, and activation of Gα12 increased the shedding of E‐cadherin. The signaling pathways of PC1 in ADPKD are not fully understood.Aim and MethodsTo understand how the interaction of PC1 and Gα12 affect the shedding of junction proteins, we used MDCK cells, Pkd1 deletion renal cells and Pkd1 knockout mice kidney tissue. Generation of inducible Pkd1 knockout mice (Mx1Cre+Pkd1flox/flox) were made by exons 2 through 6 flanked by two LoxP sites. Double knockout of Pkd1 and Gα12 mice were obtained by crossing Mx1Cre+Pkd1flox/flox mice with Gα12−/− mice. Madin‐Darby canine kidney (MDCK) Tet‐off inducible Gα12 and Gα12QL cell lines were characterized for three‐dimensional cell culture.ResultsOur results provided genetic evidence that Gα12 was essential for the development of kidney cysts induced by Pkd1 knockout, and Pkd1 deletion caused accumulation of cleaved E‐cadherin fragment in renal cystic fluid. There was no phenotype in mice with deletion of Gα12 (Gα12−/−). Conditional deletion of Pkd1 (Pkd1−/−) in one week old mice resulted in multiple kidney cysts by 9 weeks, but the mice with double knockout of Pkd1 and Gα12 (Pkd1−/−Gα12−/−) had no structural and functional abnormalities in the kidneys. In addition, Gα12 increased the active form of ADAM10 and knockdown of ADAM10 blocked the Gα12 mediated E‐cadherin shedding. Our data indicate that ADAM10 is the major sheddase for cleavage of E‐cadherin caused by Gα12 activation. Furthermore, deletion of Pkd1 increased the activation of Gα12, which subsequently decreased cell‐matrix and cell‐cell adhesion by affecting the function of focal adhesion, and E‐cadherin, respectively.ConclusionsOur results demonstrate that Gα12 activation increases ADAM10 activity and promotes the ectodomain shedding of E‐cadherin, which plays the key role in on the development of kidney cysts in Pkd1 deletion induced ADPKD.
Quantitative Proteomic Analysis of Endothelial Cells in G‐protein Coupled Estrogen Receptor (Gper1) Knockout Rats
Authors: Galla Sarah , Waghulde Harshal , Cheng Xi , Chakraborty Saroj , Mell Blair , Basrur Venkatesha , Joe Bina 
Published: November 2023

G‐protein coupled estrogen receptor (Gper1), a previously identified orphan receptor, now known to be a receptor for estrogen and aldosterone, has been localized to blood vessels, heart, and the central nervous system. Gper1 knockout models have been developed in mice and rats and have shown increased blood pressure as compared to controls. In addition, a Gper1 agonist, G1, caused decreased blood pressure when injected into mice. Women with a GPERP16L amino acid substitution in their GPER1 protein have increased BP. In order to study the effects of this receptor further, we created a novel CRISPR/Cas9 gene deletion model of the Gper1 gene on a Dahl Salt‐Sensitive (S) rat background. Hemodynamic and vascular phenotypic studies demonstrated that the Gper1 knockout rats had lower blood pressure and improved vascular function compared to the S rat. Microbiotal and metabolomics studies showed that Gper1 knockout rats had a significantly altered microbiotal composition and an increased circulating level of the short chain fatty acid, acetate. Since the improved vascular function was observed to be occurring in an endothelial dependent manner, we hypothesize that deletion of Gper1 in endothelial cells causes quantitative alterations in proteins that are directly impacted by the function of Gper1. To test this hypothesis and explore mechanisms behind the observed BP lowering effect in Gper1 knockout rats, a quantitative proteomic analysis was conducted on endothelial cells isolated and cultured from the thoracic aorta of Gper1 knockout rats and wildtype S rats. Endothelial cell cultures were digested and quantitatively analyzed by Mass Spectrometry. A total of 101 statistically significant differentially expressed proteins were identified, of which 97 proteins were with higher expression in the Gper1 knockout rats. Data prioritizing three of the most significantly upregulated proteins, Fibrillin‐1, Ephrin B1, and Cd99, as potential mechanistic links downstream of the functionality of Gper1 in endothelial cells will be presented.Support or Funding InformationFunding for this work to BJ from the NHLBI/NIH (HL020176) is gratefully acknowledged.
Inhibition of Estrogen Sulfotransferase and Adipocyte Differentiation by Hydroxylated Metabolites of Lower Chlorinated Polychlorinated Biphenyls
Authors: Parker Victoria Shayla , Squirewell Edwin , Lehmler Hans‐Joachim , Robertson Larry , Klingelhutz Aloysius , Duffel Michael 
Published: November 2023

Polychlorinated biphenyls (PCBs) are persistent molecules that are associated with the risk of developing diabetes, metabolic syndrome, cancer, thyroid abnormalities, and other diseases. PCBs are commonly found in air, food, water, paint pigments, and various materials from older buildings. The PCBs with four or fewer chlorine atoms (lower chlorinated PCBs) are readily metabolized to hydroxylated congeners (OH‐PCBs) in reactions catalyzed by cytochrome P450s. Our primary interest is in the OH‐PCBs that are derived from the lower‐chlorinated PCBs that are present in indoor and outdoor air. OH‐PCBs interact with cytosolic sulfotransferases (SULTs) as either substrates or inhibitors. Endocrine function may be altered by inhibiting the estrogen sulfotransferase (SULT1E1) that converts estradiol (active at estrogen receptors) to estradiol sulfate (inactive form), thus increasing cellular estradiol concentrations. Several of these lower‐chlorinated OH‐PCBs are potent inhibitors of SULT1E1. SULT1E1 has been previously shown to be a critical enzyme in adipogenesis, and its inhibition may alter the differentiation of pre‐adipocytes to mature adipocytes. We hypothesized that lower chlorinated congeners of OH‐PCBs that are potent inhibitors of the human SULT1E1 will also inhibit cellular differentiation in an immortalized human adipocyte cell line. Our initial studies have examined 4‐OH‐PCB 3, 4‐OH‐PCB 8, 4‐OH‐PCB 11, 4‐OH‐PCB 25, and 4‐OH PCB 52. Non‐differentiated pre‐adipocytes were treated for 4 days with either 1 μM or 10 μM of each of the OH‐PCBs, and these concentrations of OH‐PCBs did not alter overall cell viability (as judged by MTT assay). Adipogenesis studies were then conducted by incubating pre‐adipocytes in pre‐adipocyte growth media containing either 1 μM or 10 μM concentrations of each of these OH‐PCBs for 3 days. The media containing the OH‐PCB was then removed, and the cells were grown for 11 days in adipocyte differentiation media. Adipored staining was used to quantify the amount of lipid accumulation as an indicator of adipocyte differentiation. All but one of the OH‐PCBs (i.e., 4‐OH‐PCB 3) significantly inhibited adipocyte differentiation. The results were consistent both with our findings on these OH‐PCBs as inhibitors of purified recombinant human SULT1E1 and with the hypothesis that SULT1E1‐inhibition by lower‐chlorinated OH‐PCBs may inhibit adipocyte differentiation. Future work will determine cellular levels of estradiol and estradiol‐sulfate in both the pre‐adipocytes and the differentiated adipocytes under these conditions as well as changes in mRNA and protein expression for SULT1E1 and key cellular markers of differentiation.Support or Funding InformationSupported by NIH P42 ES013661 and R25 GM058939
The neurokinin‐2 receptor antagonist GR 159897 protects against neuroinflammation in the mouse enteric nervous system during colitis
Authors: Delvalle Ninotchska M , Gulbransen Brian 
Published: November 2023

Irritable bowel syndrome (IBS) is a chronic functional gastrointestinal (GI) disorder that affects 20% of Americans. The motility changes in IBS are driven by changes in the enteric nervous system (ENS). IBS has no cure, but recently, a neurokinin‐2 receptor (NK2R) antagonist has been shown to improve overall symptoms in a phase II study of diarrhea‐predominant IBS. However, the effects of NK2R antagonists on the ENS are not understood. We hypothesize that the beneficial effects of this antagonist on IBS are, in part, due to effects on the ENS. We tested our hypothesis by treating mice with the NK2R antagonist GR 159897 in a dinitrobenzene sulfonic acid (DNBS) model of colitis. To assess changes in the ENS, we used immunohistochemistry to quantify neuronal survival and performed a glial morphology analysis to assess reactive gliosis in the ENS. Calcium imaging recordings were performed in tissue preparations from mice expressing the genetically‐encoded calcium indicator GCaMP5g expressed under the control of the SOX10 promoter (SOX10::creERT2/+; PC::G5‐TdT+/−) or by loading cells with the calcium indicator dye Fluo4‐AM. Our results show that treatment with GR 159897 prevents increases in glial fibrillary acid protein (GFAP) immunoreactivity (n= 3–4 mice; p= 0.0141), increases in glial process length (n= 9–12 glia; p= 0.013) and neurodegeneration (n = 3–4; p= 0.0028). Immunohistochemical and calcium imaging data show that enteric glia express NK2Rs that are activated by neurokinin‐A (NKA). Glial responses to NKA were decreased in the presence of GR 159897 (n= 101–113 glia; p= 0.0001) or tetrodotoxin and were significantly reduced in samples from mice lacking glial connexin‐43 hemichannels (SOX10::creERT2/+; Cx43f/f mice; n= 50–113 glia; p= 0.0001). Glial responses to NKA were completely abolished in tissue samples from SOX10::creERT2/+; Cx43f/f mice in the presence of GR 159897 (n= 34–113; p= 0.0001). In conclusion, our data demonstrate that GR 159897 provides neuroprotection during colitis through mechanisms that involve decreasing glial activity driven by NKA. We speculate that these mechanisms could explain some of the clinical benefits of current NK2R antagonists in IBS.Support or Funding InformationRO1DK103723‐02S1
Regulation of Pancreatic Beta Cell Function by Ankyrin‐B
Authors: Lorenzo Damaris N , Hostettler Janell , Bennett Vann 
Published: November 2023

Functional variants of the cytoskeletal protein ankyrin‐B (AnkB) have been implicated in human hereditary cardiac arrhythmia and type 2 diabetes (T2D).1–3 We recently showed that knock‐in mice expressing human AnkB variants p.R1788W (present in 0.2% Caucasians and associated with T2D) or p.L1622I (present in 7.5% of African Americans), exhibit a metabolic syndrome that combines deficits in insulin secretion with age‐ and diet‐dependent adiposity, and insulin resistance.3 Here, we demonstrate that beta cell‐specific depletion of AnkB is sufficient to promote alterations in pancreatic islet morphology, reduction in insulin secretion, abnormal glucose tolerance, and the onset of diabetes in mice. We found that AnkB, through multiple structural and signaling functions, is a critical player in pancreatic islet biology. Biochemical and proximity ligation assays show that AnkB interacts with the inositol‐1,4,5‐triphosphate receptor (InsP3R) and with the Kir6.2 subunit of the ATP‐sensitive potassium channel. Expression of T2D‐associated AnkB mutations or loss of AnkB altered the level and localization of the InsP3R and Kir6.2 in beta cells, which, in turn, resulted in decreased insulin secretion in response to glucose and carbachol stimulation in vivo and in isolated islets, and hyperglycemia in mice. Immunohistochemistry of the pancreas revealed reduction in beta‐cell area, along with increase in apoptosis and endoplasmic reticulum stress markers. These morphological and metabolic deficits are exacerbated with age or following a high‐fat diet. These results show that Ank‐B is a critical regulator of pancreatic beta cell function in a cell‐autonomous fashion, and its deficits might cause diabetes.Support or Funding InformationHoward Hughes Medical Institute