bluuebunny/crossref_metadata_2025_split huggingface dataset:
Stimulation of intrarenal endothelin B receptors with Sarafotoxin 6c improves kidney oxygenation in rats with diabetic nephropathy by augmented oxygen availability
Authors: Franzen Stephanie , Palm Fredrik 
Published: November 2023

Diabetic nephropathy is the leading cause of end stage renal disease. The damage progression has recently been suggested to involve decreased kidney oxygenation as a result of increased oxygen consumption. Furthermore, endothelin A receptors (ETA) are highly related to kidney damage meanwhile endothelin B receptors (ETB) seem to be more protective towards diabetic nephropathy. Therefore, in this study we examined how acute in vivo intrarenal stimulation of ETB affects the progression of kidney damage in rats with diabetic nephropathy.Two weeks post induction of hyperglycemia (Streptozotocin 55 mg/kg) rats werestudied for kidney function and oxygen metabolism during acute in vivo procedures. Parameters were collected during a baseline period with intrarenal saline infusion and during an experimental period with Sarafotoxin 6c (S6c) infusion (0.78125 pmol/h).Hyperglycemic rats had lower kidney oxygenation which increased during S6c infusion. Meanwhile the increased oxygen consumption and glomerular filtration rate was unaffected by the treatment. Renal blood flow, delivered oxygen, urinary sodium excretion and oxidative stress levels increased in both groups during treatment compared to baseline.Conclusively, stimulating ETB with S6c seems to improve the diminished kidney oxygenation in diabetic nephropathy in the same manner as inhibiting ETA, by improving oxygen availability, although not to the same extent.Support or Funding InformationAll experiments were conducted according to FASEB Statement of Principles. This study was supported by the Swedish Research Council, the Swedish Diabetes Foundation and the Swedish Heart Lung Foundation.
Investigation of MicroRNA Expression Profiles in the Susceptible Population to In‐stent Restenosis
Authors: Chen Chang‐Jui , Wu Chieh‐Hsi , Chun‐Ming Shih , Huang Chun‐Yao , Bi Wei‐Fung , Chan Chao‐Shun , Kao Yung‐Ta , Pan Chun‐Hsu , Chiang Shuo‐Ju , Chiang Kuang‐Hsing , Liu Yun‐Ru , Luo Ji‐Dung 
Published: November 2023

In‐stent restenosis (ISR) is a pathological phenomenon in response to vascular injury after balloon angioplasty with stent placement, which occurs in just about 20~30% cases. Because of the lack of ISR‐specific clinical biomarkers, the drug‐eluted stents (DES) were widely abused as preventive interventions at present. A growing number of evidences suggested that abnormal expression profiles of microRNAs (miRNAs) associated with development of numerous diseases. To date, no reports showed that any miRNA candidates have been used to distinguish the population with high‐risk for ISR. Thereby, the purpose of the present study was to identify potential candidates of miRNAs differentially expressed in ISR patients. Based on the correlation network of our preliminary data from the proteiomics analysis of animal disease model and a small‐size SNP array of human samples, the candidate miRNAs with highly potential to involve in ISR development were picked up by using the MetaCore™ software. Thirty participants, including the restenosis group and the non‐restenosis group, were recruited from the Taipei Medical University Hospital, Taiwan. Total miRNA was extracted from blood serum of each participant, and expression profiles of five candidate miRNAs (mir‐142‐5p, mir‐132‐3p, mir‐24‐2‐5p, mir‐424‐5p and mir‐15b‐5p) were analyzed by using SYBR Green‐based real‐time PCR. Our data mentioned that the expression levels of two mRNAs (mir‐424‐5p and mir‐15b‐5p) showed increasing trends but another mRNAs (mir‐142‐5p, mir‐132‐3p and mir‐24‐2‐5p) were decreased in the restenosis group.Support or Funding InformationTaipei Medical UniversityTaipei Medical University Hospital
Long‐term Body Weight Regulation by The Central Melanocortin System Appears to Be Dissociated from Food Intake
Authors: Cote Isabelle , Sakarya Yasemin , Kirichenko Nataliya , Morgan Drake , Carter Christy , Tumer Nihal , Scarpace Philip 
Published: November 2023

The melanocortinergic peptide Melanotan II (MTII) is a potent appetite suppressor that rapidly promotes body weight loss when administered in the brain. Given the rapid loss of anorexic response upon chronic MTII treatment, most investigations focused on the initial physiological adaptations. However, other evidence supports the central melanocortin system as a long‐term modulator of energy balance that yet remains to be addressed. Consequently, we centrally infused either MTII (high dose; 1 μg/day or low dose; 0.04 μg/day) or artificial cerebrospinal fluid (aCSF) in 6 month‐old F344xBN rats (n = 6–7/group) for 40 days to examine the chronic effects of MTII on food intake, body weight, body composition, muscle function, and hypothalamic melanocortin receptor signaling. Central delivery of MTII suppressed appetite for five days in a dose‐dependent manner (P < 0.05). Despite that food intake promptly rose back to control level, body weight remained significantly reduced in MTII groups throughout the duration of the treatment (P < 0.01). Notably, at day 14, lean body mass was increased in rats administered aCSF relative to their respective basal value, whereas lean body mass remained stable (low dose) or decreased (high dose) in animals receiving MTII. At the end of the experiment, both MTII groups displayed approximately 40% smaller intra‐abdominal fat pads (P < 0.01) compared to the aCSF group, indicating that chronic MTII treatment targets white adipose tissues. Interestingly, although rats given MTII high dose had significantly reduced skeletal muscle mass (P < 0.05) compared to aCSF animals, neither spontaneous voluntary exercise and nor grip strength differed between groups, suggesting that MTII did not impair muscle function. At day 40, the hypothalamus of rats treated with MTII high dose was enriched with key molecules from two distinct melanocortin receptor signaling pathways. In fact, hypothalamic Phospho‐p44/42 Mitogen‐Activated Protein Kinase (P‐p44/42 MAPKs) levels of MTII high dose treated animals were six‐fold those of aCSF infused rats. Consistently, Phospho‐cAMP response element (P‐CREB) protein expression was higher (P < 0.01) in rats administered MTII high dose than in control animals. Collectively, the long‐term effects on body weight, intra‐abdominal fat mass, along with the enrichment in key signaling elements suggest that MTII induces durable physiological responses through persistent receptor activation. In conclusion, this study shows that MTII reduces body weight and intra‐abdominal adiposity through a mechanism that appears to be dissociated from changes in food intake. Our study proposes that caloric restriction efficaciously initiates body weight loss; however, combined with a secondary action that preserves a negative energy balance state over time would be a valuable approach to combat obesity.Support or Funding InformationSupported by the NIH DK091710
Active expiration and sympathetic excitation during hypercapnia require glutamatergic neurotransmission in the retrotrapezoid nucleus
Authors: Lemes Eduardo , Zoccal Daniel 
Published: November 2023

Animals exposed to hypercapnia exhibit increases in sympathetic and expiratory motor activities that are depend on, at least in part, the activation of glutamatergic neurons in the retrotrapezoid nucleus (RTN). The generation of active expiratory pattern requires the expiratory neurons of the parafacial respiratory group (pFRG), which are anatomically co‐localized with the RTN neurons. Herein we explored the hypothesis that antagonism of glutamate receptors in the RTN/pFRG prevents the emergence of active expiration and reduces the sympathoexcitation induced by hypercapnia. Microinjections of kynurenic acid (KYN, antagonist of glutamate ionotropic receptor, 100mM, 50 nL) were performed bilaterally in the RTN of arterially‐perfused decerebrated in situ preparations of Holtzman rats (60–70 g) during the exposure to hypercapnia (8% CO2) and the changes in the thoracic sympathetic (tSN), abdominal (AbN) and phrenic nerve (PN) activities were evaluated. Hypercapnia elevated tSN, PN and AbN activities of the in situ preparations (n=7). KYN microinjections in the RTN during hypercapnia reduced the sympathetic‐excitation (ΔtSN: 158±25 vs 80 ± 11%, P<0.05) and greatly attenuated the AbN response (ΔAbN: 41±13 vs 16±3%, P<0.001), but not changed the reflex increase in PN activity. Also, KYN in the RTN during hypercapnia further reduced the inspiratory time (ΔTi: −0.06±0.05 vs −0.13±0.05 s, P<0.05) and the perfusion pressure (ΔPP: −12±1 vs −20±3 mmHg, P<0.05). The results indicate that glutamatergic neurotransmission in the RTN is required for processing of the sympathetic‐excitatory and the generation of active expiratory pattern in response to hypercapnia.Support or Funding InformationSupport by FAPESP (2013/17251‐6), NIH (1R01AT008632‐01).
Improved Insulin‐Mediated Glucose Oxidation in Cultured Human Myotubes Following Roux‐en‐Y Gastric Bypass Surgery
Authors: Zou Kai , Hinkley J. Matthew , Park Sanghee , Zheng Donghai , Dohm G. Lynis , Houmard Joseph A. 
Published: November 2023

Roux‐en‐Y gastric bypass (RYGB) leads to remission of type 2 diabetes and markedly improved insulin‐mediated glucose oxidation at the whole body level, as well as in skeletal muscle. However, little is known about whether the improvements occur intrinsically at the level of skeletal muscle. The purpose of this study was to determine whether improved glucose oxidation is intrinsically retained in differentiated myotubes derived from RYGB surgery patients, and if so, the time course of the improvement. Primary human skeletal muscle cells were isolated from muscle biopsies obtained from 6 RYGB patients prior to, 1‐month and 7‐months following surgery (BMI = 50.2 ± 2.0, 43.2 ± 2.8 and 35.7 ± 2.2 kg/m2, respectively) and 6 lean subjects (BMI = 23.4 ± 0.6 kg/m2) and were differentiated to myotubes. Radiolabeled 1‐14C glucose was used to measure complete glucose oxidation (GO) and non‐oxidized glycolysis (NOG) rates in the presence or absence of insulin. Insulin‐stimulated GO rate was significantly elevated to greater extent in myotubes derived from RYGB patients at 1‐month after surgery in comparison to before surgery (7.3 ± 3.4% vs. −2.2 ± 2.3%, respectively, P<0.05). It was further enhanced at 7‐months post‐surgery (13.5 ± 2.1%, P<0.05), which was virtually restored to the level of lean people (15.4 ± 1.4%). On the other hand, NOG production in response to insulin stimulation was not significantly changed in myotubes from RYGB patients at 1‐months post‐surgery in comparison to before surgery (16.0 ± 6.2% vs. 19.5 ± 5.6%), but was largely decreased at 7‐months post‐surgery (−1.3 ± 3.6%, P<0.05), which was almost identical to the lean control group (−1.2 ± 2.5%). These changes lead to the improvement of GO/NOG ratio, an indicator of glucose partitioning, in response to insulin stimulation in myotubes from RYGB patients at both 1‐month (1.8 ± 3.8%, P=0.08) and 7‐months (11.2 ± 1.0%, P<0.05) post‐surgery when compared to before surgery (−10.8 ± 5.4%). Taken together, these data suggest that RYGB surgery can rapidly improve glucose oxidation intrinsically in skeletal muscle as early as 1‐month following surgery and further restore it to the level control level.Support or Funding InformationNational Institute of Health (DK 56112)
Transcriptomic profiling in cardiovascular disease with renal failure in human
Authors: Lee Danielle , Xu Jen , Zhang Emily , Ho Lilun , Kong Tianqing , Lu Tzongshi 
Published: November 2023

BackgroundChronic kidney disease (CKD) is a major health issue globally; it affects 50 million people worldwide and over 26 million Adults in the United States. Studies indicate that up to 50% of deaths in patients with ens‐stage renal failure are due to cardiovascular disease (CVD) and there is a 10 to 30‐fold higher cardiovascular mortality in CKD patients than in the age‐matched general population, despite adjustment for classical cardiovascular risk factors. To profile the exhaustive signatures in calcifited human endotelials, we performed whole transcriptome analysis to explore novel genes in primary human aortic endothelial cells (HAECs) and human brain microvascular endothelial cells (HBMECs).MethodsHAECs and HBECs were treated with calcification medium, and total RNA was extracted using the Nucleospin RNA isolation kit (Clontech). cDNA was synthesized and fragmented using SMART‐seq2 and Nextera XT (Illumina), respectively to generate libraries for sequencing on Hi‐seq 2000 (Illumina). Five million single‐ended raw reads from each sample were mapped to the human reference genome consortium GRCh38 by Burrows‐Wheeler Aligner (BWA); the abundancies of each gene, (FPKM, fragment per kilo‐base exon per million mapped reads), were estimated using RNA‐seq by Expectation Maximization (RSEM). Log 2 transformed FPKM were normalized by quantile normalization, which were then subjected to differential expression analysis to calculate significant fold change (Gene‐E, Broad Institute). Gene selections were performed by the combinations of fold‐changes on log 2 ratio, and p value < 0.05. Data were analyzed using Kyoto Encyclopedia of Genes and Genomes (KEGG, http://www.genome.jp/kegg/) and The Database for Annotation, Visualization and Integrated Discovery v6.7 (DAVID, http://david.abcc.ncifcrf.gov/).ResultsPathway analysis among the top 500 up‐regulated genes in comparing untreated HAECs and HBMECs to those treated with calcification medium revealed that these genes are mainly involved in cell metabolic, proliferation, and apoptosis processes. Among these genes, they were found to play roles in cell cycle regulation, apoptosis, oxidative stress, and stress resistance such as heat shock protein 70 families (HSPA1, HSPA5, and HSP40), NF‐kappa‐B inhibitor alpha, and aryl hydrocarbon receptor nuclear translocator (ARNT) in HIF‐1, NF‐kappa B and c‐AMP signaling pathways. In addition, MT‐RNR1 and cannabinoid receptor 1 are also detected that involved in amount of ATP produced in mitochondria and co‐chaperone of HSP70.ConclusionWe have identified a group of genes differentially expressed in healthy and calcified endothelial cells. Our data contribute to our understanding of the direct effects in the presence of hyperphosphatemia and hypocalcemia at endothelial cells. This gene set may have pathophysiological significance and may serve to inform rational development of therapeutic strategies to prevent CVD adverse effects in CKD patients.
Extracellular Adenosine Diphosphate Ribose Mobilizes Intracellular Ca2+ via Purinergic‐dependent Ca2+ Pathway in Rat Pulmonary Artery Smooth Muscle Cells
Authors: Huang Chun , Hu Jinxing , Subedi Krishna P. , Lin Amanda H.Y. , Sham James S.K. 
Published: November 2023

Adenosine diphosphate ribose (ADPR), a product of β‐NAD+ metabolism by the multifunctional enzyme CD38, is recognized as a novel signaling molecule. The catalytic site of CD38 orients extracellularly or intracellularly, capable of generating ADPR outside and inside the cells. CD38‐dependent pathways have been characterized in pulmonary artery smooth muscle cells (PASMCs); however the physiological function of extracellular ADPR is unclear. Here we sought to characterize the Ca2+ mobilizing and proliferative effects of extracellular ADPR and compared with the ATP‐induced responses in rat PASMCs. ADPR elicited concentration‐dependent increase in [Ca2+]i with a fast transient and a sustained phase in PASMCs. The sustained phase was abolished by Ca2+ removal and inhibited by the non‐selective cation channel blocker SKF‐96365, but was unaffected by TRPM2 antagonists or nifedipine. The IP3‐receptor inhibitor xestospongin C caused a partial inhibition of the peak and the sustained phase, while ryanodine had no effect on the ADPR‐induced Ca response. The purinergic receptor (P2X) antagonist pyridoxal‐phosphate‐6‐azophenyl‐2′,4′‐disulfonate inhibited partially the transient and the sustained Ca2+ response, while the P2(XY) inhibitor suramin and the phospholipase C inhibitor U73122 abolished the sustained Ca2+ influx. The P2Y1 antagonist MRS2179 had no effect on the response. By contrast, ATP and ADP activated Ca2+ response exhibited a high and a low affinity component, and the pharmacological profile of ATP‐induced Ca2+ response was distinctive from that of ADPR. BrdU incorporation assay showed that ADPR caused significant inhibition whereas ATP caused slight stimulation of PASMC proliferation. RT‐PCR analysis found that mRNA of most P2X and P2Y subtypes are expressed in PAs. In conclusion, ADPR and ATP activate Ca2+ responses through different combinations of multiple purinergic receptor subtypes; and extracellular ADPR may exert an autocrine/paracrine action via purinergic receptors on PASMCs.Support or Funding InformationThis work was supported by NIH grants HL071835 and HL075134,, National Natural Science Foundation of China grant NSFC31371165, and Fujian Province “Hundred Experts Award”
The collection and preservation of urinary proteins using a special diaper
Authors: Qin Weiwei 
Published: November 2023

Change is the most fundamental property of biomarker. Homeostatic nature of internal environment limits the amount and extent of changes (biomarkers) in blood, which in turn make the urine a better biomarker source (more changes) than blood, let alone that drawing blood from infants and toddlers is hard and less tolerated. For those patients with immature or no language ability, giving chief complaint is difficult. Thus, using biomarkers in urine can provide valuable clues for diagnosis of diseases especially pediatric ones. Collecting urine from little kids and some adult patients is not as easy as from healthy adults. Here we propose a method using a special diaper to collect urine. Urinary proteins were then eluted and adsorbed onto a piece of nitrocellulose membrane which can be dried and stored in a vacuumed bag. Through this procedure, the protein patterns remain faithfully preserved in SDS‐PAGE. With this simple and economical method, it is possible to collect and preserve urine samples from infants, toddlers and special patients even on large‐scale for prospective studies which are important for biomarker research on related pathophysiological conditions.Support or Funding InformationNational Basic Research Program of China (973 Program)(Nos. 2012CB517605, 2013CB530805).
Genetic tools for study of the hypothalamus and thyroid in Zebrafish
Authors: Hutcheson David , Xie Yuanyuan , Duncan Robert , Dorsky Richard 
Published: November 2023

The hypothalamus plays is a key regulatory role in a wide variety of animal behaviors including feeding, aggression, and mating. To further characterize the development and function of this CNS structure, we have begun to develop genetic tools with which to manipulate the hypothalamus in living zebrafish. Nkx2.4b is specifically expressed in the hypothalamus and thyroid during development, and is therefore an attractive target for genetic manipulation. First, we used TALENs to produce an Nkx2.4b−/− null. Damage caused by TALENs is typically repaired by non‐homologous end joining (NHEJ). We have obtained and bred several lines containing NHEJ in/dels that produce frameshifts in the 1st exon of Nkx2.4b and are predicted to be null alleles. Second, we are testing the ability of TALENS to mediate a targeted homologous recombination (HR). We created a plasmid containing Nkx2.4b genomic sequence flanking a GFP transgene, such that homologous recombination would create a “knock‐in knock‐out”. This plasmid was co‐injected along with Nkx2.4b TALENS RNAs. Initial results suggest we have successfully inserted our construct, both as a random transgene insertion as well as in a targeted manner to the Nkx2.4b locus. Confirmation by genetic mapping is underway. In nulls, the hypothalamus appears normal while the thyroid shows complete loss of both developmental and functional markers. Additionally, we seek to develop a homologous recombination vector containing a fluorophore‐2A‐CreERT2 transgene flanked by Nkx2.4b genomic sequence. This will allow equimolar expression of both a fluorescent protein as well as the CreERT2 protein. Using the same strategy we used to insert the GFP transgene into the genome, we will create a Cre driver suitable for temporally controlled manipulation of hypothalamic cells. This tool will enable a wide range of studies with the potential to greatly enhance our knowledge of hypothalamic development and function.Support or Funding InformationR01 NS082645 ‐ Regulation of hypothalamic radial glia by Wnt signaling
Molecular Cloning and Expression of a Putative Oleate Hydratase Isozyme From Nocardia Cholesterolicum NRRL 5767
Authors: Huang Jenq‐Kuen , Alhmadi Hekmat B , Vanderway David R , Hoerner Cole A , Wen Lisa 
Published: November 2023

Nocardia Cholesterolicum NRRL 5767 is well‐known for the ability to transform about 95% of added oleic acid to value‐added product, namely 10‐hydroxystearic acid (10‐HSA). Oleate hydratase is the enzyme catalyzes this conversion. The objective of this research was to clone, overexpress, and characterize oleate hydratase. Nocardia Cholesterolicum NRRL 5767 genome contains two annotated oleate hydratase. Here, we report cloning of an annotated oleate hydratase gene from the N. cholestericum genomic DNA by PCR. The gene was inserted into pET28a. Expression of functional recombinant oleate hydratase was achieved by IPTG induction. The ability to produce large quantities of the protein will permit characterization and evaluation of its suitability for use in immobilized enzyme for the production of 10‐HSA.Support or Funding InformationWestern Illinois University Foundation and the Department of Chemistry.
Optimization and Lipid Quantitation of Potential Algal Biofuels Feedstocks
Authors: Brad Samuel J. , Umbach Brynn E. , Gehlhausen Erin D. , Sweet Charles R. 
Published: November 2023

This research investigates the lipid content and composition of extremophilic and estuarine microalgae for suitability as biofuel feedstocks. These are major considerations in optimizing algal biofuel production, as they affect important fuels characteristics including production yield, melting point, flash point, cetane number, and fuel value. In these studies the known feedstock alga Chlorella saccharophila and potential new feedstocks including Galdieria sulphuraria and cold‐tolerant estuarine algae isolated from the Severn River, MD were grown mixotrophically, under replete conditions and a variety of nutrient‐depleted growth conditions. The lipid content of these organisms was determined by spectroscopic quantitation techniques in comparison with dry‐weight percentage, and acyl content was characterized by GS‐MS of fatty acid‐methyl ester (FAME) derivatives. This work compares lipid yields, fatty acid profiles, and quantitation methods from a variety of organisms under various temperature and nutrient conditions, to examine their suitability as feedstocks and determine optimal feedstock production techniques. Observations include increased relative lipid yield in Galdieria under replete conditions compared to Chlorella and increased presence of unsaturated fatty acids in the lipids of cold‐tolerant estuarine algae.Support or Funding InformationResearch Corporation Cottrell College Science Award, DoD ONR/USNA Midshipman Research Fund, and DoD DTRA Service Academy Research Initiative