bluuebunny/crossref_metadata_2025_split huggingface dataset:
Probiotics Ameliorate Gut‐Microbial Dysbiosis, Intestinal Permeability, Systemic Inflammation, and Skeletal Muscle Dysfunction in Cystathionine‐β‐synthase‐Deficient Mice
Authors: Behera Jyotirmaya , Ison Jessica , Tyagi Suresh C , Tyagi Neetu 
Published: June 2021

Recent studies have reported that dietary supplementation of probiotics is a novel approach for enhancing skeletal development and metabolism in both humans and animals. However, the precise role of probiotics in regulating skeletal muscle mass at the cellular and molecular levels in mice still needs to be clarified. Here we show that altered gut‐microbiota eubiosis, systemic inflammation and skeletal muscle dysfunction caused by hydrogen sulfide deficiency and whether the intragastric administration of probiotics (GSL#5) could retard H2S deficiency‐related skeletal muscle defects. To test this hypothesis, we employed 8‐week‐old female wild‐type (WT), cystathionine‐beta‐synthase heterozygote knockout (CBS+/−), and CBS+/− mice supplemented with probiotics (GSL#5) at the dosage of 1×109 CFU every other day through oral gavage for 6 weeks. In the murine model, H2S deficiency (CBS+/− mice) increased gut permeability, and upregulated the inflammatory cytokines TNFα (TNF), and IL‐17 in the blood and the skeletal muscle. Further, the gut microbiota is significantly altered in CBS‐deficient mice as assessed by the Denaturing gradient gel electrophoresis (DGGE) analysis. We documented that long non‐coding RNA‐PVT1 (lncRNA‐PVT1) is, indeed, upregulated in the soleus muscles of CBS‐deficient mice (4.2 fold), as assessed by RT2 lncRNA qPCR Assay. The mechanistic study shows that depressed H2S signaling stimulates the transcription factor C/EBP‐α binding to the promoter of lncRNA‐PVT1 and promotes the expression of lncRNA‐PVT1. Using an electrophoretic mobility shift assay and chromatin immunoprecipitations, we identified that lncRNA‐PVT1 interacts with TNFAIP3 interacting protein 1, TNIP1, leading to the acetylation of NF‐κB (P65 subunit) activity in the soleus muscle of CBS‐deficient mice. Further, activation of P65 region of NF‐κB transcriptionally upregulates atrogin‐1, MuRF‐1 expression. Overexpression of lncRNA‐PVT1 in cultured C2C12 muscle cell lines stimulates an increase in MuRF‐1 (2.3‐fold) and atrogin‐1 (3.7‐fold) expression. In addition, systemic delivery of synthetic lncRNA‐PVT1 in control mice causes the myofibers to undergo atrophy. Gene knockout of lncRNA‐PVT1 in CBS‐deficient mice attenuates atrophy of the myofibers and muscle wasting. Overall, administration of probiotics in CBS‐deficient mice reverses the gut‐microbiota dysbiosis and preserves skeletal muscle mass. In conclusion, our findings provide novel evidence that the administration of probiotics could balance the gut‐microbiota homeostasis as well as skeletal muscle function by modulating systemic inflammation in H2S‐deficient miceSupport or Funding InformationThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Differentiation Kinetics of Chronic Hypoxia Hematopoietic Stem Cells
Authors: Merritt Carlen Glen , Deskins Seth , Lawag Abdalla , Bacon Nick , Sollars Vincent 
Published: June 2021

In vivo, stem cells can be found in hypoxic niches within bone marrow, peripheral blood, and umbilical cord blood. Stem cells grown in hypoxic conditions, which contains an oxygen level of up to 1%, cause them to utilize glycolysis as the primary source of energy, as opposed to stem cells cultured in normoxic conditions that use both glycolysis and oxidative phosphorylation. Differentiation requires a significant amount of energy, and previous research has shown that hydroxide radicals leaked from mitochondria inhibit phosphatases and stimulate differentiation. EML cells are a stem cell factor dependent murine hematopoietic stem cell line often used to study differentiation of blood cells. In our study, EML cells were allowed to differentiate in normoxia, chronic hypoxia and acute hypoxia conditions. We observed the effects that each of the conditions have on the differentiation kinetics by using antibodies to label the cell surface markers against: Sca‐1, CD117, CD48, CD135, CD150, CD11b, CD127, F4/80, Ter119, Ly6‐G/C. Chronic hypoxia EML cells differentiation kinetics were significantly altered. Also, labeling cells with fluorescent antibodies allowed us to trace the lineage of the induced differentiated cells, by using flow cytometry. We found that EML cells cultured in chronic hypoxic conditions had a shift in its multi‐potential progenitor (MPP) subtype, MPP4 to MPP3, that the other conditions did not. Further investing the differentiation kinetics of chronic hypoxia could lead to a better understanding on producing long term hematopoietic stem cells in vitro, for stem cell transplantation in healthcare.Support or Funding InformationMy research project began November 2017 and is ongoing, and is part of Dr. Sollars' laboratory project that is funded by NIH 1R15CA186017.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Acute exposure to high‐induction electromagnetic field affects activity of model peripheral sensory neurons
Authors: Dittert Ivan , Prucha Jaroslav , Krusek Jan , Sinica Viktor , Kadkova Anna , Vlachova Viktorie 
Published: June 2021

Exposure to repetitive low‐frequency electromagnetic field (LF‐EMF) shows promise as a non‐invasive approach to treat various sensory and neurological disorders. Despite considerable progress in the development of modern stimulation devices, there is a limited understanding of the mechanisms underlying their biological effects and potential targets at the cellular level. A significant impact of electromagnetic field on voltage‐gated calcium channels and downstream signaling pathways has been convincingly demonstrated in many distinct cell types. However, evidence for clear effects on primary sensory neurons that particularly may be responsible for the analgesic actions of LF‐EMF is still lacking.Here, we used F11 cells derived from a fusion between mouse embryonic neuroblastoma and rat dorsal root ganglion (DRG) neurons as an authentic in vitro model of peripheral sensory neurons. These cells retain many features of native DRG neurons, including expression of neuropeptide receptors and various voltage‐gated channels (Vetter a Lewis, 2010, Yin et al., 2016). Using calcium‐imaging, we examined the effects of three different protocols of high induction electromagnetic stimulation on F11 chemical responsiveness and spontaneous activity. We show that short‐term (< 180 s) exposure to LF‐EMF reduces calcium transients in response to bradykinin, a potent pain‐producing inflammatory agent formed at sites of injury. Moreover, we characterize an immediate and reversible potentiating effect of LF‐EMF on neuronal spontaneous activity. Our results provide new evidence that electromagnetic field may directly modulate the activity of sensory neurons and highlight the potential of sensory neuron‐derived cell line as a physiologically relevant tool for studying the effects of LF‐EMF, and the underlying mechanisms at the cellular and molecular level.Support or Funding InformationThis work was supported by the Ministry of Health of the Czech Republic (NV16‐28784A)This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Spontaneously Hypertensive Rats is Susceptible to Bacteria Translocation from Gut to Peripheral Tissues
Authors: Zhang Bojun , Nelson James W , Bryan Robert M , Durgan David J 
Published: June 2021

Cerebral small vessel disease (CSVD) is an inflammatory disease characterized by blood‐brain barrier (BBB) breakdown and neuroinflammation. We previous observed that spontaneously hypertensive stroke‐prone rats (SHRSPs) exhibit gut dysbiosis. Furthermore, transplanting SHRSPs with control gut microbiota significantly reduced makers associated with inflammation. Since the gut microbiome is a potential source of inflammation, we hypothesize that bacteria translocating from gut to brain induce neuroinflammation in CSVD. To test our hypothesis, we determined the efficiency of fluorescent‐labeled bacteria (GFP‐E. coli JJ1901, chloramphenicol resistant) translocation in WKY and SHRSP rats. A single dose of E. coli JJ1901 [≈109 colony forming units (CFUs)] was administered to WKY and SHRSP rats (11–15 weeks old) through oral gavage. Peripheral tissues, brain, and cecum content were collected 2 hours later, homogenized, and plated on culture plates. The number of CFUs was counted 24 hours later. When compared to WKY rats, SHRSP showed significantly increased bacteria translocation to peripheral tissues (liver, spleen, and abdominal adipose tissue; n=7, p<0.05). In addition, more bacteria translocation incidents were found in SHRSPs compared to WKY rats (5/7 in SHRSP vs. 3/7 in WKY). However, similar level of acute bacteria translocation to brain was observed in both groups. To assess translocation rates of the chronically colonized bacteria, as opposed to the acute effects following gavage, colonization was established in newborn rat pups by administrating E. coli JJ1901 (≈109 CFUs) to pregnant mother rats through oral gavage. Bacteria colonization in the offspring of the gavaged rats was confirmed by plating diluted fecal samples on culture plates. Colonized SHRSPs (15 weeks old) showed significantly increased bacteria translocation to peripheral tissues when compared to WKY controls. Additionally, SHRSPs demonstrated a trend towards increased translocation of E. coli JJ1901 to the brain than WKY rats. We conclude that translocation of gut bacteria can be an underlying source responsible for inflammation in pathological states.Support or Funding InformationNational Heart, Lung, and Blood Institute R01HL134838 (D.J. Durgan), and by National Institute of Neurological Disorders and Stroke R01NS080531 (R.M. Bryan)This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Cholesterol crystals increase vascular permeability by inactivating SHP2 and disrupting adherens junctions
Authors: Mani Arul Murugan , Singh Nikhlesh K. , Rao Gadiparthi N. 
Published: June 2021

To understand the adverse effects of cholesterol crystals on vascular homeostasis, we have studied their effects on endothelial barrier function. Cholesterol crystals increased endothelial barrier permeability in a dose and time‐dependent manner. In addition, cholesterol crystals induced tyrosine phosphorylation of VE‐cadherin and a‐catenin, disrupting endothelial AJ and its barrier function and these effects required xanthine oxidase‐mediated H2O2 production, SHP2 inactivation and Frk activation. Similarly, feeding C57BL/6 mice with cholesterol‐rich diet increased xanthine oxidase expression, H2O2 production, SHP2 inactivation and Frk activation leading to enhanced tyrosine phosphorylation of VE‐cadherin and a‐catenin, thereby disrupting endothelial AJ and increasing vascular permeability. Resolvin D1, a specialized pro‐resolving mediator, prevented all these adverse effects of cholesterol crystals and cholesterol‐rich diet in endothelial cells and mice, respectively. Based on these observations, it is likely that cholesterol crystals via disrupting AJ increase vascular permeability, a critical event of endothelial dysfunction and specialized pro‐resolving mediators such as Resolvin D1 exert protection against these effects.Support or Funding InformationHL074860 from NHLBI to GNR.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Age‐ and Salt‐dependent Changes in miRNA Profile in FBN rat and C57BL/6J mice: Role of sodium hydrogen exchanger regulatory factor‐1 (NHERF1)
Authors: Jain Anish , Aubee Joseph I , Thompson Karl M , Knepper Mark A , Lee Dexter L , Khundmiri Syed Jalal 
Published: June 2021

Previous studies from our laboratory demonstrated decreased NHERF1 expression in aging F344. Loss of NHERF1 expression prevented dopamine‐dependent inhibition of NKA in renal PTCs from aged F344. Our laboratory demonstrated increased expression of miRNAs regulating markers of inflammation in NHERF‐1 deficient mice and rat models. We hypothesized that loss of NHERF1 expression prevents salt‐sensitive hypertension due to changes in miRNA expression regulating Na‐transporters in renal tubules. To address this hypothesis, we analyzed the miRNA expression profile in 4mo and 24mo old FBN rats and 18mo old C57BL/6J wild‐type and NHERF1 knock‐out mice fed a normal salt or 8% salt diet (HSD). Total RNA from kidney cortex was isolated using Trizol (miRVana, Ambion) and miRNA profile was analyzed using kits and software from Nanostring. The top 100 miRNAs with cut‐off set at counts three times the SD of the negative control miRNA counts were considered for analysis. Counts from normal diet fed animals (n=8, 4M4F) were averaged together and considered as 1 to calculate changes in HSD animals. In 4‐mo old FBN rats 23 miRNAs significantly increased in HSD (miR196c, miR466b, miR144, and miR142‐5p), while miR547, miR375, and 6 others significantly decreased. In 24‐mo old FBN rats 30 miRNAs increased significantly in HSD (miR‐192, miRNA141, and miR21), while 10 miRNAs decreased significantly (miR328a and miR‐let‐7a and f). Similar results were observed in WT mice. However, in NHERF‐1 KO mice the miRNAs increased strongly regulate markers of inflammation. The data was confirmed by measuring the expression of down‐stream targets using western blotting and ELISA. These data may suggest that NHERF1 plays a major role in regulation of renal inflammation and salt‐sensitive hypertension.Support or Funding Information7R21AG047474‐03 NIH; R25AG047843‐NIH; 16GRNT31030019‐AHAThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The increased expression of microRNAs 451, 638 and 362 in Urinary Exosomes of Human Subjects profiled as Diabetic and Hypertensive
Authors: Lewis Harvey , Smith Michael , Fluitt Maurice , Khundmiri Syed J , Ecelbarger Carolyn , Lee Dexter L 
Published: June 2021

Urinary Exosomes (UE) are involved in several metabolic processes, including hypertension and are considered to be vehicles for eradicating cell waste products. UE have important functions in human diseases as carriers of information including, proteins, lipids and miRNA (mi‐R). Because of their high stability, UE mi‐Rs can be useful predictors of the progression of diabetes and hypertension. UE mi‐Rs 451, 638, 362 and 16 are potential biomarkers for early diabetic nephropathy, salt‐sensitive hypertension and cardiovascular disease, respectively. We hypothesized that mi‐Rs 451, 638, 362 and 16 are increased in diabetic + hypertensive patients when compared to controls. To address this hypothesis, human urine samples were collected from control, diabetic and hypertensive patients. UE were isolated in each group and mi‐R 451, 638, 362 and 16 expression was quantified by RT‐PCR. mi‐R expression was expressed as log2, normalized to U6 small nuclear RNA as mean and standard error. UE mi‐R 451 expression in diabetic + hypertensive patients was 2.4 ±1.8 and 0.5 ±0.8 in controls. UE mi‐R 638 expression in diabetic + hypertensive patients was 1.0 ±2.0 and 0.1 ±1.7 in controls. UE mi‐R 362 expression in diabetic + hypertensive patients was 1.9 ±1.8 and 0.2 ±0.8 in controls. UE mi‐R 16 expression in diabetic + hypertensive patients was similar to the controls (0.1 ±10 and −9 ±1, respectively). Our results suggest that both mi‐R 451, 638 and 362 expressions are increased in diabetic + hypertensive patients. There was not an increase in UE mi‐R 16 in diabetic + hypertensive patients when compared to controls. In conclusion, mi‐R 451, 638 and 362 expressions may serve as non‐invasive biomarkers for the early detection for diabetes and hypertension.Support or Funding InformationR25AG047843‐NIHThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Differential Nuclear Localization of SOX18 Variants in Transiently Transfected Epithelial Cells
Authors: Cvammen William , Prokop Jeremy W. , Qutob Dinah , Freeland Thomas , Underwood Adam 
Published: June 2021

Sry‐related box (SOX) proteins are transcription factors containing a conserved High Mobility Group‐box (HMG‐box) DNA‐binding domain that regulate embryogenesis and tissue maintenance. SOX mutations result in cancer, developmental abnormalities, sex reversal, and circulatory dysfunction. Genomic analysis of the 20 human SOX genes identified 3,049 variants from gnomAD and 1,566 from COSMIC with an enrichment of variants that falls within the HMG box. The most impactful variant identified occurred in the third alpha helix of the SOX18 HMG‐box that replaces a glutamic acid (E) for a lysine (K) (rs201931544). Since other variants occurring within the HMG box of SOX18 are linked to vascular and lymphatic disfunction, we predict SOX18 E137K will also derange protein function through altered DNA interaction and localization to the nucleus. No literature exists regarding E137K, although it is present in 0.8% of Latino population. The purpose of this project was to produce native and mutated SOX18 expression constructs to determine if nuclear accumulation is impacted. Native human SOX18 was first cloned into pHTCHaloTag® CMV‐neo vector (Promega), followed by site‐directed mutagenesis to synthesize pHTCHalo/SOX18‐E137K. After sequence confirmation, human dermal lymphatic endothelial cells, HeLa and A375 were transiently transfected with either the native or mutated construct, or a control vector (pHTCHaloTag® CMV‐neo vector with no insert), in glass chamber slides. After 24hr, live cells were incubated with HaloTag® Oregon Green® Ligand (Promega) and micrographs were captured on an Olympus iX51 with DP71 digital camera. Controls showing cytochemistry specificity include cells transfected with SOX18 incubated with PBS in place of Oregon Green® Ligand. From these trials, it was determined that E137K reduces nuclear localization of SOX18 in all cell lines and may account for divergent phenotypic consequences when compared to native SOX18.Support or Funding InformationSupport for this study was from NIH‐K01ES025435 (to JWP) and Walsh UniversityThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The current unrecognized epidemic of smell dysfunction in the United States
Authors: Henkin Robert 
Published: June 2021

There is an estimated 21 million people with smell dysfunction in the United States. However, most people in the U.S. are unaware of the existence of this problem. Most physicians are also unaware of this problem. Indeed, there is no specific medical specialty to evaluate or treat this problem. These issues comprise an unrecognized epidemic of smell dysfunction occurring in the U.S. Smell dysfunction comprises patients with loss of smell acuity, the presence of distorted perception of external odors (aliosmia) and the presence of distorted smells in the nose in the absence of any odor (phantosmia). These symptoms most commonly occur after a viral‐type illness (about 25%), allergic rhinitis (about 20%), head injury (about 15%) and a variety of other illnesses including Parkinsonism, Alzheimer disease and other degenerative neurological disorders. The current view of the pathology which generates these symptoms is that some neurological abnormalities occurred either in the olfactory nerves or in the brain. However, recent studies have demonstrated that the underlying cause of these symptoms is not neurological in nature but usually biochemical related to loss of olfactory receptor function due to inhibition of biochemical activation and signaling in stem cells upon which olfactory cell growth and development depend. The recognition that biochemical treatment of this stem cell pathology results in activation of specific sensory cell signaling which improves smell dysfunction demonstrates the biochemical basis of this unrecognized epidemic.Support or Funding InformationThis institution did not receive any funding from agencies in the public, commercial, or not‐for‐profit sectors.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Gut Microbiome Composition Differs between Individuals with Acute and Long‐standing Traumatic Spinal Cord Injury
Authors: Li Jia , Hoover Thomas , Morrow Casey , VanDerPol William , Yarar‐Fisher Ceren 
Published: June 2021

ObjectiveTo examine the gut microbiome composition among individuals with acute and long‐standing spinal cord injury (SCI).MethodsSeven individuals with acute spinal cord injury (A‐SCI, 36 ± 13 y, 2F/5M, C4‐T10, AIS A‐D) and 10 individuals with long‐standing SCI (L‐SCI, 49 ± 9 y, 1F/9M, C4‐T12, AIS A, B, and D ) were recruited. Stool samples were collected after a median of 7 days after the onset of SCI (range: 4–11 days) from participants with A‐SCI in the hospital. The median number of years post injury for the L‐SCI group was 22 years at the time of stool collection. Parapak® vials was used for collection of the stool samples. Fecal microbiota DNA was extracted using Zymo® Fecal DNA Isolation kit. PCR was used to amplify the V4 region of the 16S rRNA gene, and the PCR products were sequenced using an Illumina MiSeq platform. Bioinformatics analyses started with assessment of raw data and filtering low‐quality data using FASTQC and FASTX, respectively. A combination of tools in the QIIME suite were utilized for clustering reads into operational taxonomic units (uclust) and taxa assignment (PyNAST and Fasttree). Several metrics for alpha and beta diversity were determined for group comparisons. Significant differences between groups (A‐SCI vs L‐SCI) were determined by PERMANOVA test on each of the beta diversity indices. A Kruskal‐Wallis test was performed to compare the abundances of key taxa between groups using tools within the QIIME suite.ResultsMicrobiota alpha diversity was not significantly different between groups. However, group differences were observed in beta diversity (unweighted and weighted UniFrac analyses, P < 0.05 for both). Principle coordinate analysis showed a separation of two groups (see attached figure). Individuals with chronic SCI had higher abundances of bacteria in the Clostridiaceae, Lachnospiraceae, and Alcaligenaceae families, lower abundances of S24‐7 family, and higher abundances of Parabacteroides genus (family Porphyromonadaceae) (P < 0.05).ConclusionThese data for the first time demonstrated a sustained shift in gut microbiota composition in individuals with L‐SCI vs. A‐SCI. Specifically, microbes linked to the development of metabolic syndrome, central nervous system‐related bowel disorders, and physical inactivity are elevated in adults with L‐SCI. Future studies are warranted to evaluate the involvement of gut microbiota in the development of metabolic disorders among individuals with L‐SCI.Support or Funding InformationThis work was supported by KL2TR001419‐01 (CY‐F) and H133N110008 (McLain A)This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.