bluuebunny/crossref_metadata_2025_split huggingface dataset:
Proof of Principle: Transfer of Small Numbers of Bacteria from Mother to Fetus
Authors: Yu Kevin , Zarate Miguel , Keller‐Wood Maureen , Wood Charles E 
Published: June 2021

The fetus is thought to develop in a sterile environment in utero. Long standing dogma that “the uterus and the feto‐placental unit is “sterile” is based primarily on microbiological culture‐based techniques that were unsuccessful in growing “culture resistant” bacteria or intracellular bacteria. We have reported the presence of low numbers of bacteria in tissues from normal sheep fetuses in pregnancies not complicated by infection. The exposure of the fetus to bacteria might aid neonatal survival by informing fetal immune development. We propose that the fetus is not sterile and that bacteria or fragments of bacteria can be transferred from mother to fetus. We hypothesize that inoculation in the maternal mouth results in the appearance of bacteria in the fetus. We used S. aureus containing green (GFP), red (RFP), or far‐red (FRFP) fluorescent protein‐expressing plasmids to inoculate late‐gestation pregnant sheep (gestational age= 130–135 days, n=7) intravenously (GFP, 104 cfu), into maternal mouth (RFP, 104 cfu) and vagina (FRFP, 104 cfu). These were small doses of bacteria which did not cause physiological (no fever) or psychological (no anorexia) signs of infection. Five to seven days post maternal inoculation, animals were humanely sacrificed, and fetal tissues were collected, and DNA was extracted from placenta and fetal liver, spleen, and cerebral cortex. We probed for GFP plasmid using several primer pairs for endpoint PCR. While detection of whole‐length plasmids was not successful, PCR reactions probing for smaller fragments of plasmid were successful. We found GFP plasmid DNA in all tissues in 10/10 fetal livers and RFP in 10/10 livers. The appearance of GFP and RFP‐labelled bacteria in fetal liver (p=7.497e‐06) was statistically significant as tested by Chi‐Square analysis. We did not detect significant FRFP plasmid DNA in liver. Analysis of tissues by immunohistochemistry revealed GFP and RFP‐expressing bacteria in fetal tissues, although most appeared to be in aggregates. We conclude that S. aureus, introduced in small numbers in maternal mouth and bloodstream, appear in the fetus and placenta. We were unable to determine whether these bacteria were alive in the fetus.Support or Funding InformationThis work was supported by HD033053, AI120195, and HL083810This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Expression of Genes that Comprise the Core Molecular Clock are Altered in the Atrophied Skeletal Muscle by Androgen Deprivation
Authors: Rossetti Michael L , Casero Robert A , Esser Karyn A , Gordon Bradley Scott 
Published: June 2021

Skeletal muscle atrophy increases the risk of morbidity and mortality during various pathological conditions. In males, a decrease in the production and/or bioavailability of androgens (termed hypogonadism) directly contributes to muscle atrophy during various pathological conditions. While it is known that androgens prevent muscle atrophy, the mechanism(s) by which androgens mediate this effect are largely undefined. Our laboratory previously showed that mitochondrial turnover is enhanced in the tibialis anterior (TA) muscle of mice by androgen deprivation induced by castration surgery, and the magnitude of turnover was related to the degree of muscle atrophy. These data suggest that potentially dysfunctional mitochondria contribute to the muscle atrophy observed following androgen deprivation. To gain a better understanding of that factors that might contribute to changes in mitochondrial quality control during androgen deprived conditions, we subjected total RNA from the TA of sham and castrated mice to microarray analysis. This unbiased approach identified significant changes in expression of genes that comprise the core molecular clock. qRT‐PCR confirmed that expression of Brain and Muscle Arntl 1 (Bmal1) was decreased, while expression of Period 1, Period 2, and Period 3 (Per1, 2, & 3) were increased in the TA of castrated mice. When measured across a diurnal cycle, the change in expression of Bmal1, Per1, and Per2 exhibited reduced amplitude under androgen‐deprived conditions. Interestingly, strong relationships were observed between the castration‐mediated changes of core clock components and the measures of mitochondrial turnover. Specifically, Bmal1 expression was directly related to BCL2/adenovirus E1B 19 kDA protein‐interacting protein 3 (BNIP3) protein content (R2 = 0.88), and the expression of core clock components were also directly related to the content of various mitochondrial proteins. Expression of core clock components were also related to the autophagy marker, p62, and the mass of the TA. Ex post facto analysis of the microarray also identified changes in genes regulating polyamine biosynthesis. As polyamines are known to alter core clock function, we determined whether androgen deprivation altered polyamine content. While castration did not alter Spermine content, there was a significant reduction in the content of Spermidine in the TA of castrated mice. Overall, these data suggest that reduced Spermidine concentrations may contribute to alterations in the core molecular clock in the skeletal muscle under androgen‐deprived conditions, which may in turn contribute to reduced mitochondrial quality control and subsequent muscle atrophySupport or Funding InformationNIH‐CA204345 (RAC) and Institute for Successful Longevity (BSG)This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Effect of short‐term 17B‐estradiol on low‐flow‐mediated constriction in young women
Authors: Kuczmarski Andrew , Sebzda Kelly , Wenner Megan 
Published: June 2021

Low‐flow mediated constriction (L‐FMC) examines vascular responses during a reduction in blood flow and provides a measure of resting shear‐stress dependent vascular tone. L‐FMC is a complementary measure to flow‐mediated dilation (FMD), and is blunted in patients with cardiovascular disease. 17β‐estradiol (E2) exerts positive effects on vascular endothelial health and can improve FMD. However, it is unknown whether E2 impacts L‐FMC.PURPOSEThe purpose of this study was to test the hypothesis that E2 administration would augment L‐FMC in young women.METHODSEleven young women (24±1 years, 24±1 kg/m2) have completed the study. To isolate the impact of E2, endogenous ovarian hormone production was suppressed with daily subcutaneous injections of a gonadotropin releasing hormone antagonist (GnRHant, 0.25 mg/day) for 10 days. E2 (0.1mg/day patch) was administered during the last 7 days of GnRHant. We measured changes in brachial artery diameter via ultrasound on day 3 of GnRHant and day 7 of E2 add‐back. A blood pressure cuff was placed just distal to the olecranon process and inflated to 200mmHg for five minutes. Images were recorded continuously at baseline (1‐minute), during occlusion (5‐minutes), and after cuff deflation (2‐minutes). L‐FMC was determined from the last 30 seconds of cuff inflation using the lowest average value and was reported as percent change from baseline diameter. FMD was calculated as a percent change from baseline to peak diameter after cuff deflation. Total vessel reactivity was calculated as the absolute value of L‐FMC and FMD. Results are reported as mean ± standard error.RESULTSBaseline diameter was similar between GnRHant (3.14 ± 0.12 mm) and E2 (3.08 ± 0.11 mm; p=0.3). L‐FMC tended to be greater during E2 administration (GnRHant −2.5 ± 0.5% vs. E2 −3.8 ± 0.5%; p=0.11). However, FMD (GnRHant 7.5 ± 0.7% vs. E2 7.6 ± 1.1%; p=0.92) and total vessel reactivity (GnRHant 10.0 ± 0.7% vs. E2 11.5 ± 1.2%; p=0.26) were not altered by E2.CONCLUSIONThese preliminary findings suggest that short term E2 administration may augment L‐FMC in young women. Additional data are needed to understand the impact of ovarian hormones on L‐FMC and its role in regulating vascular health.Support or Funding InformationNIH Grant P20 GM 103446, P20 GM 113125, U54 GM 104941, The University of Delaware Research Foundation, and AHA Award 16SDG30700015This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Maternal Overconsumption of High Fructose Corn Syrup Causes Hypoxia in Placenta and Fetal Livers of Wistar Rats
Authors: Ahmed Erum , Carrillo‐Sepulveda Maria Alicia 
Published: June 2021

BackgroundAmerican maternal overnutrition negatively impacts fetal development. Several epidemiological and experimental studies have demonstrated that overconsumption of diets rich in fats and sugars can have effects spanning three generations. While the negative impact of high fructose corn syrup (HFCS), a component heavily present in many processed American foods, is well‐recognized, its effects during fetal development remain largely underexplored. Thus, this study aims to examine the placental and fetal hepatic effects of HFCS maternal overconsumption.MethodsAdult female Wistar rats were randomized in 2 experimental groups: HFCS Group received a 20% HFCS solution in drinking water for 30 days prior to and during pregnancy. Control Group received regular water. Prior to pregnancy, blood pressure was measured in both experimental groups by using tail‐cuff plethysmography. Fasting glucose readings were measured weekly. Terminal experiments were completed on day 21 of gestation. Placentas and fetal livers were isolated and further processed for analysis of Hypoxia‐Inducible Factor‐α (HIF‐α) expression, a classical transcription factor of hypoxia, using Western Blot. Blood samples were collected for biochemical analysis.ResultsPrior to pregnancy, HFCS group exhibited increased serum triglycerides (441.48 vs. 109.47 mg/dL), systolic (159.03 ± 12.04 vs. 136.29 ± 2.57 mmHg, p<0.05) and diastolic blood pressures (114.66 ± 9.47 vs. 99.29 ± 2.49 mmHg, p<0.05) compared to controls. No differences were found in glucose levels (98.17 ± 1.65 vs. 92.5 ± .24 mg/dL controls). At the terminal experimental protocol, HFCS group displayed increased HIF‐α expression in placenta (2.3 fold‐increase, p<0.05) and fetal liver (3.1 fold increase, p<0.001) compared to controls. Therefore, our results show hypoxic changes in both placental and fetal hepatic tissues. Moreover, oil red staining confirmed increased fat deposition in the maternal livers from HFCS group.ConclusionOur findings show that overconsumption of HFCS prior to and during pregnancy has a negative impact on offspring, as demonstrated by upregulation of hypoxic markers such as HIF‐α. We will further this study to determine whether other tissues undergo hypoxia and the mechanisms by which HFCS leads to hypoxia in maternal and fetal tissues.Support or Funding InformationIn‐house Grant NYITThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
10‐Week High‐Fat Diet Promotes Endotoxemia and Alters Microbial Taxonomy in Male Adolescent Rats
Authors: Crawford Meli'sa Shaunte , Sweazea Karen 
Published: June 2021

Diet‐induced changes in gut microbiota are associated with increased risk of obesity and diabetes. Prior research shows that a proprietary soil‐derived compound decreases weight gain and blood glucose in diabetic mice. The aim of this study was to examine the effects of a similar organometallic complex (OMC) on gastrointestinal inflammation in rats fed high‐fat diet (HFD). Six‐week‐old male Sprague‐Dawley rats (n=36) were divided into two dietary groups: chow (18.9% protein, 57.33% carbohydrates, 5% fat) or HFD (20% protein, 20% carbohydrates, 60% fat) for 10 weeks. Animals were further divided (n=6/group) and administered 0, 0.6 or 3.0 mg/mL OMC in their drinking water. Microbial taxonomy of cecal contents were evaluated by LEfSe analysis and showed a significant abundance of Rikenellaceae, Alistipes, Erysipelotrichaceae and Oceanospirillales in samples from chow rats (LDA <2.0) after 10 weeks. In contrast, HFD cecum contents were more enriched in bacteria classified as “other” and “mitochondria” (LDA <2.0). HFD rats also showed elevated lipopolysaccharide concentrations (p<0.05), which was prevented by OMC. Additionally, HFD rats showed an increase in fat accumulation in the liver as indicated by elevated triglyceride concentrations (p<0.05). Furthermore, plasma ALT activity was increased in HFD rats (p<0.05). Western blot analyses of the cecum showed no changes in protein expression of the inflammatory cytokines NF‐kB, IL‐1B, or IL‐6 (Two‐Way ANOVA, p=0.362, p=0.702, and p=0.746, respectively) in rats fed HFD. Likewise, protein expression of NF‐kB, IL‐1B, and IL‐6 were not altered (Two‐Way ANOVA, p=0.402, p=0.062, or p=0.953 respectively) by OMC‐treatment. Thus, while a 10‐week HFD induces gut dysbiosis and endotoxemia, gut inflammatory biomarkers were not significantly altered by diet or OMC. Further studies are needed to determine whether OMC alters intestinal permeability.Support or Funding InformationThis study was partially funded by a grant from Isagenix International, LLC.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The Taste Receptor Gene Repertoire in Salmonids
Authors: Gomes Ana Silva , Lai Floriana , Taché Yvette , Rønnestad Ivar 
Published: June 2021

In vertebrates, G protein–coupled receptors (GPCRs) detect the sweet, umami and bitter tastes. The sweet and umami GPCRs are encoded by the T1R gene family, whose members heterodimerise to detect molecules associated with sweet (T1R2‐T1R3) and umami (T1R1–T1R3) tastes. The bitter receptors are encoded by the T2R gene family. In mammals, these receptors detect not only nutrients and toxins in the oral cavity but also in the gastrointestinal (GI‐)tract. Here, these receptors are located in enteroendocrine cells facing the lumen that trigger in response to nutrients the release of hormones, enabling the control of many gut physiological functions.The number of T1R genes is highly conserved among different species except for some lineage‐specific changes, whereas the numbers of T2R genes diverge drastically across the vertebrate lineage. In this context, the additional and relatively recent whole‐genome duplication (WGD) event in the salmonids (salmonid‐specific 4th WGD) offers an excellent model system to study the evolution and adaptation of the taste receptor genes, their functional specialization and their possible role in gut nutrient‐sensing system.In this study, we analyzed the genome database of five salmonid species, Salmo (S.) salar, Oncorhynchus (O.) tshawytscha, O. mykiss, O. kisutch and Salvelinus (S.) alpinus, and found that only 2 to 4 t2r genes are present in salmonids species, similar to what has been observed for other teleost species (with the exception of Astyanax mexicanus). In O. mykiss and S. alpinus 6 t1r genes were identified, 7 were found in the S. salar and O. kisutch and 9 genes in the O. tshawytscha genoma. The number of t1r3 genes has expanded in all salmonids species (4 genes were found in the O. tshawytscha genome and 2 genes in the other species) compared to most other teleost species (with the exception of carps which also undergone a 4th round of WGD). Phylogenetic and short‐linkage analysis should elucidate the taste receptor gene evolution dynamics in salmonids and across the vertebrate lineages. In addition, we are investigating the mRNA expression levels of the t1r and t2r in S. salar. Preliminary data suggests that t1r transcripts are abundantly expressed in the oropharyngeal area and some t1r transcripts may be present in specific regions of the GI‐tract. This could confirm that the functional role of T1R and T2R in sensing nutrients and bitter components in the GI‐tract observed for mammalian species also exist in the salmonid lineage, however further studies are necessary to proof this hypothesis.Support or Funding InformationFunded by the Research Council of Norway (NFR 262096 GUTASTE).This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Identifying and Characterizing Hepatic Sensory Receptors
Authors: Kurtz Ryan , Steinberg Lily G , Shepard Blythe D 
Published: June 2021

Sensory receptors, including olfactory receptors (ORs), taste receptors (TAS), and orphan G‐protein coupled receptors (GPRs) have recently been found in a variety of non‐sensory tissues where they have important physiological functions. As G protein‐coupled receptors (GPCRs), these receptors serve as chemosensors by interpreting and responding to chemical cues in their native environment (i.e. blood, urine) and can be activated by naturally occurring metabolites. We reasoned that the liver, the largest metabolic organ in the body, is primed to take advantage of some of these sensory receptors in order to regulate its internal environment. To identify the complete complement of sensory receptors expressed in murine liver, we performed an unbiased screen of whole liver using TaqMan qPCR array cards. This screen identified a total of 16 ORs, 10 TAS, 3 opsins, and 57 GPRs – with 7 of the GPRs expressed at levels equal to well‐characterized hepatic GPCRs. In order to understand the functions of these sensory receptors, it is imperative to determine where they localize within the liver and their ligand profile. Unfortunately, due to the lack of reliable antibodies for most sensory receptors, localizing these receptors and resolving their binding profiles can be challenging. To begin, we focused initially on the hepatic ORs. Using gene‐specific primers and RT‐PCR, we successfully confirmed the presence of 16 ORs consistently expressed in male and female murine liver. The majority of the identified hepatic ORs are orphan receptors with no known ligands. This is due, in part, to the inability to functionally express ORs in heterologous cell systems. To examine functional expression of Olfr99 and Olfr267, the two highest expressing liver ORs, we cloned the full length ORs from the liver into an expression system containing an N‐terminal Flag tag and expressed them in HEK293T cells. Using live‐cell surface labeling for the extracellular Flag epitope, we were able to identify conditions that promoted functional expression of Olfr99, but not Olfr267, using a cleavable LUCY tag as well as two chaperone proteins, RTP1s and Ric8b. With expression conditions established, we began a large‐scale ligand screen using a cAMP driven Dual‐Luciferase assay. To date, we have screened Olfr99 against a total of 49 compounds and mixtures including small odorant molecules that cover a wide odorant space and physiologically‐relevant molecules including hepatic metabolites and bile acids. Although a strong activator has yet to be found for Olfr99, our results have uncovered the identification of novel sensory receptors in the liver. Efforts are currently underway to determine the localization and physiological function of these receptors to better understand how the liver responds to its immediate environment.Support or Funding InformationNIDDKThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Differential DNA Methylation Between African Americans And European Americans: the Role of Childhood Socioeconomic Status and Relations to Health Disparity
Authors: Pan Yue , Wang Xiaoling , Treiber Frank , Harshfield Gregory , Su Shaoyong 
Published: June 2021

RationaleDifferent DNA methylation patterns between ethnic groups may reflect the impact of social and environmental factors and contribute to health disparity. Increasing evidence indicates the importance of low socioeconomic status (SES), particularly in childhood, as a partial explanation for poor health outcomes in African Americans (AAs) compared to European Americans (EAs). This study aims to identify ethnicity‐related methylation patterns and their relationship with health disparities, and further determine to what extent the identified signals could be explained by potential difference in childhood SES between AAs and EAs.MethodsGenome wide DNA methylation data from leukocytes of 224 AAs (58% females) and 244 EAs (46% females), aged 18–37, were obtained from Georgia Stress and Heart study using the Infinium HumanMethylation450 BeadChip. Childhood SES was assessed using parental Hollingshead score.ResultsBy comparing AAs and EAs, 1671 differentially methylated CpG sites (DMCs) (p<1×10−7) were identified. Gene ontology analysis using DAVID showed that the genes harboring these DMCs were significantly enriched (FDR<0.05) in disease terms of chem dependency, tobacco use, body weight, cardiovascular, metabolic and developmental disorders, which were complex traits with known racial disparities. Furthermore, the ethnicity‐related DMCs showed significant enrichment in DMCs identified by epigenome‐wide association studies for obesity (p<7×10−28), hypertension (p<0.01), and type 2 diabetes (p<2.4×10−5), supporting the role of ethnicity‐related DMCs in explaining the health disparity in metabolic health between AAs and EAs. In addition, AAs showed significant lower childhood SES compared to EAs (37.8±13.4 vs. 45.2±12.2, p<0.001). Moreover, among the 1671 ethnicity related DMCs, 766 were significantly associated with childhood SES (FDR<0.05). Mediation analysis further revealed that childhood SES could partially explain the ethnicity differences in DNA methylation levels in 193 out of the 766 DMCs (mediation effects: 7%–22%, p<0.05).ConclusionsWe found significant differences in DNA methylation levels between AAs and EAs, and these differences were highly enriched with respect to DMCs related to metabolic traits, suggesting that epigenetics underlying the ethnicity difference may partially explain the health disparity in metabolic health between AAs and EAs. We also found that ethnicity related differential methylation may be partially mediated through disproportionately lower childhood SES in AAs relative to EAs.Support or Funding InformationHL125577, HL069999This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.