bluuebunny/crossref_metadata_2025 huggingface dataset:
A novel mouse model of kidney fibrosis induced by glomerulus‐specific overexpression of the full‐length of human transforming growth factor‐β1 gene
Authors: Takeshita Atsuro , Yasuma Taro , Hozumi Kenta , Nishihama Kota , D’ Alessandro‐Gabazza Corina N. , Totoki Toshiaki , Toda Masaaki , Yano Yutaka , Gabazza Esteban C
Published: June 2021

Chronic renal disease is a common complication of several diseases including diabetes mellitus, hypertension and chronic glomerulonephritis. Approximately 10% of adults in developed countries suffer from chronic renal disease. Clinical progression of chronic renal disease leads to end‐stage renal disease that can be treated only with renal replacement therapy including dialysis and renal transplantation. There is currently no other therapeutic option for patients in terminal stages of the disease. The main reason of the scarce therapeutic strategies for this chronic and fatal disease is the lack of appropriate disease model to evaluate drug discovery. In the present study, we focused on the potent profibrotic activity of transforming growth factor‐β1 and its critical mechanistic role in glomerulosclerosis. We prepared and amplified a chimeric mouse podocin/human transforming growth factor‐β1 bacterial artificial chromosome construct, purified, linearized, separated by pulsed field gel electrophoresis and dialyzed before microinjection into fertilized eggs. We obtained several founders expressing the transgene as demonstrated by Southern blotting. As expected, the transgenic mice have increased plasma levels of human transforming growth factor‐β1, creatinine and blood urea nitrogen from 8‐weeks of age. The urine levels of fatty acid‐binding protein, a marker of kidney injury, and human transforming growth factor‐β1 were also significantly increased in the transgenic mice compared to wild type mice. Trichrome staining of the kidneys disclosed extensive deposition of extracellular matrix proteins in the transgenic mice compared to their wild type counterparts. In addition to highlighting the important role of transforming growth factor‐β1 in the pathogenesis of kidney fibrosis, the results of this study led to development of a novel mouse model of kidney fibrosis that may be useful for drug discovery.

Support or Funding Information

I have no financial relationships to disclose.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Effects of Resistance Exercise on Kidney Antioxidant Enzymes of Rats Treated with Doxorubicin
Authors: Busekrus Raquel , Hydock David Scott , Tigner Allison , Kaufenberg Mikayla , Hook Sarah , Bredahl Eric
Published: June 2021

The aim of this study was to investigate the effects of the anti‐cancer drug doxorubicin (DOX) and resistance exercise training on kidney antioxidant enzyme expression. Male rats were assigned to a resistance training (RR) or sedentary (SS) group for the first 10 weeks of the study. RR were housed in specialized resistance training cages, and SS were house in standard cages. After 10 weeks, animals received either weekly 3 mg/kg intraperitoneal injections of DOX over 4 weeks (12 mg/kg total, SSD, RRD) or an equivalent doses of 0.9% saline as control and continued with the activity treatments (SSS, RRS). Five days after the last DOX injection, kidneys were collected, and expression of the antioxidants catalase (CAT), superoxide dismutase 1 (SOD1), and glutathione peroxidase (GPx) was analyzed using Western blotting. Significant activity and drug main effects were observed for CAT expression (p=0.02 and p=0.0002, respectively) with SSD expressing significantly lower CAT than all other groups (p<0.05). With SOD1 expression, a significant activity effect (p=0.0088), drug effect (p<0.0001), and activity × drug interaction (p=0.0347) was observed. No significant main effects or interactions were observed for GPx, but there was a trend toward an activity effect (p=0.0706). Although more work is needed, the results of this study are promising, and we conclude that resistance training prior to and during DOX treatment may help to minimize renal damage by upregulating antioxidant enzyme expression.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The Effect of Fennel (Foeniculum vulgare) on the JAK/STAT Pathway and Intestinal Barrier Function
Authors: Rabalais John , Kozan Philip , Lu Tina , Durali Nassim , McGeough Matthew D. , Marchelletta Ron , Lee Jae Beom , Barrett Kim , Sivagnanam Mamata
Published: June 2021

Foeniculum vulgare (F. vulgare), commonly known as fennel, is a flowering plant species in the family Apiaceae. It is believed to be one of the world's oldest medicinal herbs and has been used for its reported anti‐inflammatory and antipathogenic properties. Anethole, a major component of F. vulgare, has also been shown to inhibit tumor necrosis factor (TNF)‐induced activation of nuclear factor kappa B (NF‐kB) and interleukin‐1β in rats both of which are parts of wide ranging inflammatory pathways. F. vulgare has also been used as a complementary and alternative treatment for intestinal bowel disease (IBD) in some countries. To investigate this alternative treatment and F. vulgare's anti‐inflammatory properties further, the Janus Kinase/Signal Transducers and Activators of Transcription (JAK/STAT) pathway, which is known to be active in many forms of IBD, was selected to be interrogated by growing colonic (T84) monolayers in vitro with F. vulgare oil extract before being challenged with interferon‐gamma (IFN‐γ). A dextran sodium sulfate (DSS) murine model that was given F. vulgare extract was also used to model ulcerative colitis.

Barrier studies of T84 monolayers showed pretreatments with F. vulgare oil extract significant protected transepithelial electrical resistance (TEER) as compared to monolayers which only received vehicle treatments prior to IFN‐γ challenge (Fig. 1A, P<0.01). IFN‐γ has been shown to significantly decrease TEER in cells.? Furthermore, tight junction protein‐1 (TJTP‐1) and occludin (OCLD), both important tight junction proteins, showed increased RNA expression in F. vulgare treated monolayers (Fig. 1B–C, P<0.05). Levels of STAT1 mRNA and total STAT1 protein were similar between IFN‐γ treatments, regardless of F. vulgare treatment (Fig. 2A, P>0.05). However, F. vulgare did inhibit phosphorylation of STAT1, showing decreasing amounts of pSTAT1 with increasing amounts of F. vulgare in T84 monolayers (Fig. 2B, P<0.05). In the DSS‐murine studies, mice given F. vulgare treatments showed marked decreases in ulcer indices as compared to mice only given DSS (Fig. 3B, P<0.05). mRNA transcripts of STAT1 showed decreases in F. vulgare treated mice when compared to mice only given DSS (Data not shown). Both phosphorylated STAT1 and total STAT1 levels were decreased in F. vulgare treated mice as compared to mice only given DSS (Fig. 4, P<0.05). The results of the in vitro and in vivo experiments demonstrate F. vulgare's protective effects on gastrointestinal barrier integrity and overall macroscopic structure in the colon. We propose that F. vulgare inhibits phosphorylation of STAT1 in vitro and inhibits both phosphorylation and transcription of STAT1 in vivo.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Engineered bacterial Tight Junctions: a high‐throughput method to identify absorption enhancers
Authors: Rollings Jay , Worthington Tyler , Mizrachi Dario
Published: June 2021

Tight Junctions are the master regulators of the paracellular space and the gatekeepers of the blood‐tissue barriers. Claudins are the membrane proteins responsible for cell‐cell contacts at the Tight Junctions. Absorption enhancers (AE) are molecules that disrupt the paracellular space at the Tight Junction level, allowing for increased transport of material between cells. Thus, greatly enhancing the translational potential for drug delivery and even for enabling new drug discovery. Currently, no AE have been identified for clinical use because the current setting of studying Tight Junctions involves mammalian cell monolayers in a very low throughput setting. We engineered a claudin protein which can be expressed in the outer membrane of E. coli. Thus, the E. coli are endowed with Tight Junction‐like structures driving cell to cell interactions. This new approach provides us with a high‐throughput method to interrogate investigate large libraries of compounds to identify AE. Our results indicate that known AE affect both mammalian and E. coli Tight Junctions in similar fashion. Results from the high‐throughput E. coli system can be translated to mammalian cells and result in candidates for translational solutions and new drug design.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Effect of Univestin (Scutellaria baicalensis root & Acacia catechu heartwood) on Post‐Exercise Muscle Soreness and Range of Motion in Healthy Adults
Authors: Talbott Shawn M , Talbott Julie A , Hantla Don
Published: June 2021

Introduction

Intense exercise is known to increase post‐exercise pain and result in delayed‐onset muscle soreness (DOMS) lasting for several days. Muscle soreness and soft tissue pain is a frequent complaint among recreational exercisers and a significant obstacle to continued participation in regular physical activity.

Objective

Our study examined the effects of an herbal dietary supplement (Univestin; combination of Scutellaria baicalensis root & Acacia catechu heartwood) versus a look‐alike placebo on subjective pain and range of motion (ROM) following intense exercise.

Subjects & Methods

Twenty‐three (N=23) healthy men and women were recruited to participate in an intense 60‐minute “boot camp style” resistance training workout designed to induce DOMS across all major muscle groups (quadriceps, hamstrings, gluteals/hips, shoulders, biceps, triceps). Subjects were randomly divided in double‐blind fashion to receive either the supplement (Univestin) at 250 mg b.i.d. (N=12) or placebo (N=11) for 1‐week post‐exercise. We measured range of motion (ROM), joint pain (WOMAC; Western Ontario & McMaster Universities Osteoarthritis Index), and muscle pain/stiffness (VAS; visual analog score) daily during the 1‐week post‐exercise period.

Results

Subjective evaluation of pain/stiffness (VAS) was noticeably lower across several muscle groups (calves, gluteals/hips, biceps, triceps) at day‐1 and significantly lower (p<0.05) by the 2nd day post‐exercise in the Univestin group compared to Placebo. ROM scores were significantly higher across both shoulder and hip movements (flexion/extension) at all time points from day‐1 to day‐6 post‐exercise in Univestin compared to Placebo (p<0.05). Measures of joint pain (WOMAC) were also lower in Univestin versus Placebo at some, but not all, timepoints.

Conclusion

Univestin has previously been reported to reduce the pain and stiffness associated with osteoarthritis (1–3). These clinical results further demonstrate Univestin's effectiveness to reduce post‐exercise muscle pain and improve range of motion in a population of healthy non‐arthritic adults. Therefore, Univestin may be viewed as an effective dietary supplement to facilitate healthy pain‐free participation in regular physical activity.

Support or Funding Information

Funding provided by Unigen

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Biological Metabolite and Resveratrol Interaction in Cholesterol Diet Fed Rabbits
Authors: Jimoh Abdulazeez , Olusegun Ayorinde Joseph , Tanko Yusuf , Abubakar Ahmed , Aliyu Mohammed
Published: June 2021

Serum adiponectin concentration is inversely associated with leptin and insulin concentration. There is little paucity of information in the literature on the adipocytes levels and regulations by resveratrol in cholesterol diet‐fed rabbit. Thirty rabbits were randomly divided into six groups, each of five animals: Group 1 = control, Group 2 = cholesterol diet (CD) only, Group 3 = resveratrol 200 mg/kg (R200), Group 4 = resveratrol 400 mg/kg (R400), Group 5 = CD + R200 and group 6 = CD + R400. The preparations were administered for eight weeks of experimental protocol. weight and blood glucose level were measured using digital weighing balance and glucometer on week zero and the eighth week of the experiment. At the end of the study period, the rabbits were placed under light anesthesia (ketamine) to remain unconscious. blood sample of about 3ml was drawn via cardiac puncture for serum extraction which were evaluated for serum concentration of metabolites assays (adiponectin, leptin and insulin). Blood glucose level and body weight were significantly higher in CD group compared to groups that received CD + resveratrol supplement and resveratrol only. Serum adiponectin showed significant (P < 0.05) increased in CD + resveratrol groups, compared to CD group only. Leptin and insulin significantly shows (P < 0.05) decreased in in CD groups co‐administered with resveratrol and resveratrol groups only compared to CD group only.in conclusion, the decrease in body weight and blood glucose level demonstrated the antidiabetic effect of resveratrol and increase in adiponectin level, decrease in leptin and insulin level following resveratrol supplement elucidated the therapeutic potential of resveratrol, despite cholesterol diet consumptions.

Support or Funding Information

TETfUND (Tertiary Education Trust Fund Nigeria) and Ahmadu Bello University, Zaria, Kaduna State, Nigeria

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Fenofibrate Prevents Weight Gain in DIO Mice Independent of Subcutaneous White Adipose Tissue Browning
Authors: Rodrigues Alice Cristina , Rocha Karina Cunha
Published: June 2021

Activation of peroxisome proliferator activated receptor alpha (PPARα) affects energy metabolism in adipocytes. Our group and others have shown fenofibrate, a PPARα agonist, increases energy expenditure and prevents weight gain in diet‐induced obese mice. The aim of this study is to evaluate if fenofibrate‐induced weight loss is a consequence of subcutaneous white adipose tissue browning. Male C57BL/6 mice at 8 weeks' old (n ± 40; CEUA 165/2011) were randomly assigned to receive a control (CD) or a high‐fat diet (HFD) for 6 weeks. After that, they were subdivided into four groups: C group (fed with CD, untreated), CF group (fed with CD and treated with fenofibrate), H group (fed with HFD, untreated) and HF group (fed with HFD and treated with fenofibrate). Fenofibrate (50 mg/Kg/day) was administered daily by oral gavage for 2 weeks. After 48h of the last dose, they were euthanized. Expression of mRNAs (Ucp1, Elovl3 and Eva‐1) in inguinal adipose tissue were measured by Real‐time PCR assay. Mice fed with HFD showed augment of body weight gain, visceral fat depots weights, fasting leptin and insulin serum levels and reduced constant of glucose disappearance (Kitt). Without changes in food intake, fenofibrate treatment attenuated the weight gain in both CF and HF groups, contributing to reduce the adipose tissue storage, and restored insulin and leptin levels when compared to C group. Kitt was not different between HF and H groups. Expression of Ucp1, Elovl3 and Eva‐1 were down regulated in obese mice and fenofibrate had no effect on their expression. On the other hand, in control mice, fenofibrate increased Eva‐1 expression, a gene characteristic of classical murine brown fat. In conclusion, fenofibrate prevents weight gain in obese mice by a mechanism that seems not to involve subcutaneous white adipose tissue browning.

Support or Funding Information

FAPESP and CAPES

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
SLN overexpression upregulates mitochondrial dynamics and promotes oxidative metabolism during caloric restriction.
Authors: Periasamy Muthu , dos Reis Felipe Gomes
Published: June 2021

Obesity results from increased caloric intake, physical inactivity and is a major contributor to type 2 diabetes (T2D). Exercise and caloric restriction (CR) are the most effective interventions to prevent obesity, T2D and sarcopenia. By increasing fatty acid oxidation (FAO), CR promotes insulin sensitivity and improves mitochondrial function. Our laboratory has identified Sarcolipin (SLN) as an important player in muscle thermogenesis. SLN‐binding to SERCA promotes uncoupling of Ca2+ transport, causing futile cycling of the SERCA pump, increasing energy expenditure. SLN overexpression in glycolytic muscle leads to increased mitochondrial biogenesis and FAO, resembling an oxidative phenotype. Therefore, we sought to determine if SLN overexpression (1) promotes resistance to fatigue and increased exercise performance and (2) improves the beneficial effects of CR in skeletal muscle mitochondrial metabolism. Wild type (WT), Sarcolipin knockout (SLN−/−) and Sarcolipin overexpression (SLNOE) mice were submitted to 8 weeks of exercise training at 60% of maximal effort capacity. After exercise, oxygen consumption, maximal force and resistance to fatigue were measured in isolated muscles. Electronic transport chain (ETC) and oxidative phosphorylation (OXPHOS) related proteins were analyzed in muscle and adipose tissue. Exercise‐induced increase in ETC proteins of white adipose tissue was blunted in SLN−/− mice. Our results show that SLNOE are resistant to fatigue and show better functional recovery as compared with WT and SLN−/−. Oxygen consumption is also increased in tibialis anterior after exercise in SLNOE, indicating that SLN overexpression in glycolytic muscle improves FAO and promotes endurance capacity. To induce CR, 3 month‐old male WT, SLN−/− and SLNOE mice were submitted to 8 weeks of 40% CR protocol, and body weight was measured weekly. Glucose tolerance test and insulin sensitivity measurements showed that SLNOE improved CR‐induced insulin sensitivity and glucose tolerance. Western blot analysis revealed that CR‐induced increase in ETC proteins (Complex I to V) and mitochondrial content (Tfam) is SLN‐dependent. Secondly, SLN increases mitochondrial fusion (Mfn1 and Opa1) and fission (Drp1) proteins and, as expected, CR increased markers of autophagy (LC3‐ II). Our data indicates that SLN plays an important role in metabolic adaptations in muscle by modulating mitochondrial dynamics and promoting oxidative metabolism. We propose that SLN can contribute to CR induced metabolic benefits in muscle.

Support or Funding Information

NIDDK ‐R01 DK098240

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Adipocyte Specific Expression of the PI3 Kinase Regulatory Subunit p85α Causes Reduced Insulin Sensitivity in Mice
Authors: Hetrick Byron , Clayton Zach , Chick Wallace S. , McCurdy Carrie
Published: June 2021

Obese adipose tissue is characterized by reduced insulin response and elevated, persistent inflammation. Obese adipose tissue has also been shown to have increased abundance of the regulatory subunits of Phosphoinsoitide 3‐kinase (PI3 kinase), p85α, p55α, and p50α. Preventing the upregulation of the PI3 kinase regulatory subunits in response to high fat diet improved insulin response and reduced inflammation in white adipose tissue. p85α contains unique N‐terminal adaptor regions that allows its function outside of regulation of PI3 kinase localization and activity. We therefore hypothesize that p85α upregulation plays a unique role contributing to insulin resistance in obese adipose tissue. To understand how adipocyte specific increases in p85α influence insulin response, we have generated a mouse model overexpressing the PI3 kinase regulatory subunit, p85α, in an adipocyte specific manner. Expression of Cre recombinase is under control of the adiponectin promoter, restricting its expression to white and brown adipocytes. Cre mediated recombination results in overexpression of p85α under control of the constitutive ROSA26 promoter. We observe overexpression of p85α specifically in white adipose tissue. In oral glucose tolerance tests, mice overexpressing p85α show an increased glucose area under the curve. Reduced adipocyte insulin response, or increased inflammation caused by adipocyte p85α overexpression may contribute to the reduced glucose tolerance. We are investigating how adipocyte specific p85α overexpression influences adipose tissue insulin response through modulation of PI3 kinase/Akt signaling pathway in response to insulin. Increased p85α expression may alter PI3 kinase activity, in addition, p85α has been shown to stabilize and activate Phosphatase and tensin homolog (PTEN), which acts in opposition to PI3 kinase to attenuate the PI3K/Akt signal transduction pathway. Besides the direct interactions in the insulin signal transduction cascade, blocking the obesity induced upregulation of PI3 kinase regulatory subunits attenuated the pro‐inflammatory environment of obese adipose tissue. We therefore also predict that p85α overexpression will increase the pro‐inflammatory gene expression profile of adipose tissue through currently unidentified pathways. Based on our observations in an adipocyte specific p85α overexpression mouse line, we propose that high fat diet induced upregulation of p85α is a pathogenic rather than adaptive response to overnutrition. Blocking high fat diet induced upregulation of p85α would likely be an effective strategy to attenuate high fat diet induced insulin resistance.

Support or Funding Information

NIH DK095926‐01

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Mitochondrial DNA Damage And Impaired Iron Homeostasis In Muscle Aging
Authors: Picca Anna , Mankowski Robert , Calvani Riccardo , Marzetti Emanuele , Leeuwenburgh Christiaan
Published: June 2021

Iron dyshomeostasis has been indicated as a major risk factor for the development and progression of several disease conditions. Impairment of iron balance is also an aging feature especially in post‐mitotic tissues. In particular, while age‐related systemic iron deficiency (anemia) compromise oxygen availability, increased cellular and mitochondrial iron levels have been involved in DNA mutation load.

We purified total proteins and DNA from muscle samples of young (Y, n=10) and sedentary older adults, classified as high‐ (HFE, n=14) and low‐functioning (LFE, n=6) based on the Short Physical Performance Battery (SPPB). The effect of aging on key proteins involved in iron homeostasis and qualitative alterations of mtDNA and how these relate to physical performance was evaluated.

A decrease in muscle volume, as quantified through 3D‐NMR, was found in LFE (38%) and HFE (30%) participants compared to the younger counterpart and positively correlated with physical performance. A lower expression of the iron import protein transferrin receptor (TfR1) but not Zip14 was also found in both groups of elderly participants, regardless of the SPPB score. The analysis of the mutation load of mitochondrial DNA (mtDNA) (e.g., single‐strand, double‐strand breaks and abasic sites), instead, showed an increase in mtDNA damage in HFE and LFE participants, with no differences between the two groups. This study showed an association between altered mitochondrial homeostasis and levels of intracellular iron in aged human muscles. The accumulation of mtDNA mutations and the decline in iron balance may therefore represent critical steps to muscle aging and possible targets for interventions against muscle wasting.

Support or Funding Information

This study was supported by the National Institute on Aging (NIA) (R01 AG17994), the Claude D. Pepper Older Americans Independence Center (1 P30 AG028740), “Centro Studi Achille e Linda Lorenzon”, and Fondazione Roma (NCDs Call for Proposal 2013).

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
A Review‐ Structure‐Function studies of Abrin: A ribosome inactivation toxin
Authors: Singh Sarabjit
Published: June 2021

Abrin is a highly toxic protein produced by Abrus precarious, with an estimated human fatal dose of 0.1–1 μg/kg, and has caused death after accidental and intentional poisoning. Abrin can be extracted from jequirity beans using a relatively simple and cheap procedure. Abrin from the seeds of Abrus precatorius is type II ribosome‐inactivating proteins. Abrin induces apoptosis in cells by triggering intrinsic mitochondrial pathway. The toxins inhibit protein synthesis probably by inactivating the 60S ribosomal subunits, interfering with chain elongation. Only eukaryotic ribosomes are attacked by the toxin. Toxin resistance involves an impairment of the transfer of the toxin A‐chain from the cell surface to the target, the ribosomes. A remarkable group of proteins challenge the notion that protein sequence determines a unique three‐ dimensional structure, and that membrane and soluble proteins are very distinct. Protein synthesis inhibition by abrin and agglutinin‐I in two different cell lines and found that 200–2000 fold higher concentration of agglutinin‐I is needed for the same degree of inhibition. The description of the abrin‐ A chain, APA‐I A chain consists of three folding domains. Domain 1 is composed of residues 2–108 consisting of six stranded β‐sheet, one β‐hairpin and two helices. Domain 2 consists of residues from 109–196 and is the most conserved part of the A chain with total five helices. Domain 3 consists of residues 197–261. It consists of two helices and two anti‐parallel strands. Abrin consists of an A‐chain of 251 amino acids preceded by 34 amino acids followed by a 14 amino acid linker and a B chain of 263 amino acids. There is not more research on abrin in India. Therefore i try to study about toxicity, function and structure of abrin protein.

Support or Funding Information

Self funded.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Developmental Nicotine Exposure Alters Upper Airway Protective Reflexes In Neonates
Authors: Clarke Jordan , Cross Seres , Fregosi Ralph
Published: June 2021

Although smoking while pregnant is associated with developmental abnormalities in newborns, at least 20% of women smoke during pregnancy. In addition, many women that quit smoking in pregnancy are prescribed nicotine replacement therapies, despite limited knowledge of its developmental consequences on the fetus. Human infants that were smoke‐exposed in utero tend to have more obstructive apneas during sleep than unexposed infants. Here we report the results of experiments designed to examine the impact of developmental nicotine exposure (DNE) on motor output to upper airway muscles in neonatal rats. Pregnant dams were exposed to nicotine (6 mg/kg of nicotine tartrate daily) with an osmotic mini‐pump implanted subdermally on the 5th day of gestation. Neonates (P3‐P10) were lightly anesthetized and placed in a head out body plethysmograph to measure airflow, and fine wires were inserted into the genioglossus muscle to measure the EMG. We examined ventilatory and EMG responses during and after 15‐sec nasal occlusions. The change in EMG burst amplitude in response to nasal occlusion was blunted in DNE animals compared to control, but only in animals younger than P7. DNE animals also show an increased latency period (time of first EMG burst after the onset of nasal occlusion) compared to control animals, again only in animals P7 and younger. Twelve of the 112 animals studied failed to recover following nasal occlusion, and 9 of those 12 were nicotine‐exposed. These results show that nicotine exposure in‐utero does alter upper airway protective reflexes in neonates by increasing the time it takes to respond to respiratory stress and blunting the magnitude of each effort to breathe during their response. Moreover, DNE animals were more likely to die following a nasal occlusion challenge.

Support or Funding Information

NIH RO1HD071302

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Endothelin‐1 and COMP Regulate Heme Biosynthesis and Guanylate Cyclase Expression in Smooth Muscle of Pulmonary Arteries
Authors: Alruwaili Norah , Yu Hang , Kelly Melissa R. , Sun Dong , Wolin Michael S.
Published: June 2021

The objective of this study is to examine if heme biosynthesis is regulated in smooth muscle of pulmonary arteries by endothelin‐1 (ET1) and/or conditions modulating the availability of Cartilage Oligomeric Matrix Protein (COMP). Our recent studies in endothelium‐rubbed bovine pulmonary arteries (BPA) organoid cultured for up to 48 hours observed ET1 (10nM) increases superoxide in the mitochondrial matrix and extra mitochondrial regions associated with COMP depletion. While incubation with COMP (0.5μM) attenuated ET1‐induced superoxide increases. We hypothesize that increased superoxide in the mitochondrial matrix could impair heme biosynthesis from protoporphyrin IX (PpIX) by decreasing the availability of Fe2+ and/or ferrochelatase (FECH). To examine this hypothesis, we studied the effects of ET1, COMP and siRNA depletion of COMP on the expression of FECH, transferrin receptor‐1 (TfR1, an indicator of iron availability) and soluble guanylate cyclase (sGC, a key heme‐dependent protein), and on measurements of PpIX (HPLC) and heme content. ET1 decreased FECH, heme and sGC, and increased PpIX and TfR1. COMP reversed these effects of ET1, and COMP also decreased PpIX and increased heme in the absence of ET1. The depletion of COMP by siRNA increased detection of mitochondrial Matrix (MitoSOX‐HPLC) and extramitochondrial (DHE‐HPLC) superoxide, PpIX and TfR1 expression, and decreased the expression of FECH, sGC and the phosphorylation of VASP on the site mediated by cGMP protein kinase. In vascular reactivity studies, it was observed that relaxation of BPA to the nitric oxide donor spermine NONOate was inhibited by organoid culture with ET1, and this effect of ET1 was attenuated in the presence of COMP during the exposure to ET1. Collectively, ET1 and COMP depletion appears to impair the biosynthesis of heme in pulmonary arteries potentially associated with the loss of protective sGC and cGMP vasodilator and remodeling‐inhibitory mechanisms.

Support or Funding Information

Supported by NIH Grants HL115124 & HL129797; The Natural Science Foundation of the P. R. China (31400989); The Natural Science Foundation of Heilongjiang Province (C2015069) ; Postdoctoral Science Foundation of Heilongjiang Province (LBHQ15087)

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Long‐term Hypoxia Alters Cytochrome P450 Activity During Pregnancy in Ovine Species
Authors: Truong Hillary , Lam Tran Chau , Solak Sara , Adeoye Olayemi Olufikayo
Published: June 2021

The absolute avoidance of medication during pregnancy, though desired, might not be practical because of preexisting co‐morbidities, pregnancy induced disorders (e.g. gestational diabetes), preeclampsia, or even fetal disorders. When indicated, the use of medication could be a major source of stress and apprehension for the pregnant woman and her care‐provider. The inadequacy of data on drug safety during pregnancy is one contributory factor to this apprehension. Compounding this problem is the resurgence of the use of illicit drugs and abuse of prescription medications such as opioids during pregnancy. It is well‐established that drug metabolism is altered during pregnancy. Changes in maternal drug metabolism can also occur during pathologic conditions associated with exposure to varying levels of hypoxia.

Cytochrome P450 (CYP) 3A accounts for the metabolism of over 60% of clinically used drugs and environmental xenobiotics. CYP expression is altered by pregnancy as well as pathologies attributable to hypoxia like eclampsia, placental insufficiency and illicit drug use. We hypothesize here that long term hypoxia (LTH) alters CYP activity in pregnant ovine species via epigenetic mechanisms. To generate the hypoxic experimental groups, non‐pregnant and pregnant sheep were taken to an altitude of 3280m above sea level for 110 days while normoxic controls for each group were kept at sea level. Post sacrifice, precise cuts of liver segments were cleaned of blood and assayed for CYP3A4 activity and total methylated cytosines. Compared to their normoxic controls, LTH increased hepatic CYP3A4 catalytic activity in non‐pregnant sheep (118%) and pregnant sheep (142%). Whereas hepatic CYP3A4 catalytic activity was not significantly different between non‐pregnant and pregnant normoxic sheep, it was decreased in pregnant hypoxic sheep (20%) compared to non‐pregnant hypoxic sheep. Western blot quantification revealed no significant changes in CYP3A4 protein abundance between normoxic vs. hypoxic and non‐pregnant vs. pregnant sheep. Lastly, quantified total methylated cytosines (TMC) showed that, compared to their normoxic controls, LTH increased the TMC in non‐pregnant sheep (65%) but decreased (48%) the TMC in pregnant sheep. TMC was increased (38%) in pregnant compared to non‐pregnant normoxic sheep but decreased (56%) in pregnant compared to non‐pregnant hypoxic sheep. Together, these results suggest that gene silencing via CpG methylation of candidate CYP genes may contribute to changes in the catalytic activity of CYP3A4 during pregnancy but not non‐pregnant states.

Support or Funding Information

Loma Linda University School of Pharmacy Faculty Scholarship Fund.

NIH Program Project Grant from NICHD (HD31266) and an NIH R01 Grant from NINDS (NS076945).

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Differences in Peak VO2 Among Healthy Andean Highlanders and Males with Chronic Mountain Sickness Before and After Isovolemic Hemodilution at 4350m
Authors: Heinrich Erica C. , Anza‐Ramirez Cecilia , Tift Michael S. , Marcarlupu Jose Luis , Wagner Harrieth E. , Figueroa‐Mujica Romulo , Vizcardo‐Galindo Gustavo , Gu Wanjun , Wagner Peter D. , Villafuerte Francisco C. , Simonson Tatum S.
Published: June 2021

Excessive erythrocytosis is the hallmark of Chronic Mountain Sickness (CMS), a maladaptive clinical syndrome common in Andeans that is thought to negatively impact oxygen transport and exercise capacity. We performed cardiopulmonary exercise testing at 4350m in both healthy males (controls) and those with CMS pre‐ and post‐isovolemic hemodilution (n = 8 controls, n = 6 CMS) to determine whether they differed in exercise capacity as a function of hematocrit (Hct). Hct differed significantly between control (Hct=51.6±4.0) and pre‐hemodilution CMS subjects and between pre‐ and post‐hemodilution (Hct=66.7±4.1, 53.2±1.8, respectively; all p<0.0005), and Hct showed a negative trend with exercise capacity in CMS subjects. Anaerobic threshold was lower in CMS subjects compared to controls (p<0.01) but normalized after hemodilution. While peak VO2/kg was similar pre‐ and post‐hemodilution in CMS subjects, it was significantly different between control and CMS subjects with or without hemodilution (p<0.05). Cardiac output (QT/kg) during maximal exercise was lower in pre‐ and post‐hemodilution CMS compared to controls (p<0.02) and was relatively lower in pre‐ versus post‐hemodilution CMS subjects (p<0.05). The slopes of the QT‐VO2 relationship indicate that decreased QT response to exercise in CMS subjects is partially normalized post hemodilution (p<0.04), while the intercept is not statistically different. While QT is reduced in CMS and can be partially recovered by hemodilution, VO2 max did not increase in CMS individuals post hemodilution, suggesting this reduction of Hct does not influence exercise capacity. How the differences between CMS and non‐CMS individuals relate to other components of oxygen transport and the presence and absence of adaptive genetic factors remains to be determined.

Support or Funding Information

This study was supported by an NIH NRSA Individual Postdoctoral Fellowship (Parent F32) to ECH (F32HL131218), a Wellcome Trust PHTM grant (107544/Z/15/Z) to FCV, and an NIH R00HL118215 and American Physiological Society Giles F. Filley Memorial Award to TSS.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Heme‐oxygenase 2 (HMOX2) variants associated with evolutionary adaptation and hemoglobin concentration in Tibetans are common in Andean Highlanders
Authors: Yip Gigi , Heinrich Erica C. , Villafuerte Francisco C. , Simonson Tatum S.
Published: June 2021

Andean and Tibetan populations have developed distinct adaptations to high altitude. The heme oxygenase 2 gene (HMOX2) is under selection in Tibetans and is associated with hemoglobin concentration in Tibetan men. Putatively adaptive variants within this gene are also prevalent in lowland population but have not been examined in Andean highlanders and could influence variation in hypoxia responses in this population. We genotyped ten variants within the HMOX2 gene region (rs2058812, rs2270363, rs6500606, rs3747584, rs4786509, rs860891, rs8129, rs11639998, rs9934267, and rs17761816) in 128 Quechua Andean highlanders in Cerro de Pasco, Peru (4,350 m) (female = 61; male = 67). We found that the frequencies in Andean highlanders are not as elevated as Tibetans' but similar to those observed in Han Chinese. After controlling for sex and age, there was no significant association between these SNPs and hematocrit (Hct), mean arterial pressure (MAP), oxygen saturation (SpO2), nor hypoxic ventilatory response (HVR). These results suggest that the putatively adaptive high‐altitude HMOX2 variants identified in Tibetans are found at intermediate frequencies in Andeans, although they may not be regulators of hemoglobin concentration in this population.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Autophagy‐Related Protein 16L1 (Atg16L1) Depletion Induces Insulin Resistance Through Decreased IRS Expression
Authors: Frendo‐Cumbo Scott , Jaldin‐Fincati Javier , Brumell John H , Klip Amira
Published: June 2021

Autophagy is a degradation pathway persisting in all cells at basal levels and further activated by various stimuli, such as starvation. This cellular process is mediated by autophagy‐related (Atg) proteins, which govern cargo recognition and formation of a nascent organelle called the autophagosome. Several Atg proteins have been identified, including Atg3 and the Atg12‐Atg5‐Atg16L1 complex. Recently, connections between autophagy and insulin action have emerged. The insulin receptor (IR) and insulin‐like growth factor receptor (IGF1R) bind and phosphorylate insulin receptor substrate 1 (IRS1) leading to PI3K/Akt activation, which through downstream activation of mTORC1 inhibit autophagy. On the other hand, tissue‐specific, autophagy‐deficient mouse models have attenuated basal and insulin‐stimulated PI3K/Akt signalling. Similarly, expression of the dominant‐negative mutant ATG5K130R in L6 myoblasts reduces insulin‐induced Akt and IRS1 phosphorylation. As growth factor signalling reduces autophagy, these findings suggest a potential feedback mechanism whereby deficient autophagy acts to attenuate insulin signalling. However, the mechanism through which reduced autophagy feeds back onto insulin signalling remains unknown. We hypothesize that interfering with autophagy affects insulin and IGF1 signalling at different levels, including IR, IGF1R and IRS1, and we investigate the potential underlying mechanism. To examine the interplay between deficient autophagy and insulin action, we treated mouse embryonic fibroblasts (MEFs) depleted of Atg3, Atg5, Atg12 or Atg16L1 with 10nM insulin or 100ng/ml (13.3nM) IGF1. Atg depletion did not induce changes in IR content, its localization at the plasma membrane or insulin binding. Interestingly, Atg16L1‐KO MEFs have reduced IRS1 and IRS2 protein content and gene expression, which suggests that defective Atg16L1 affects IRS gene transcription. ER stress is known to reduce Foxo1 protein content, an important transcription factor of IRS, and we have found elevated ER stress in Atg16L1‐KO MEFs. Accordingly, Atg16L1‐ and Atg5‐KO MEFs exhibited attenuated insulin‐ and IGF1‐induced Akt phosphorylation at both S473 and T308 compared to WT. In contrast, insulin‐ and IGF1‐induced Akt phosphorylation was inhibited selectively at T308 in Atg3‐KO MEFs, and at S473 in Atg12 KO MEFs. In conclusion, Atg16L1 appears to induce insulin and IGF1 resistance through decreased IRS1 and IRS2 expression, while Atg3 Atg5 and Atg12 affect signalling at the level of Akt.

Support or Funding Information

SFC is funded by NSERC‐CGS.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Distinct Heat Stress Responses of Porcine Cumulus Cells and The Oocyte Is Associated With Disrupted Intercellular Communication
Authors: Yin Chao
Published: June 2021

Cumulus cells (CCs) play critical roles in oocytes maturation through intercellular communication. The adverse effect of heat stress (HS) on oocyte maturation has been well documented, whereas the HS responses of CCs and the oocytes in association with cumulusoocyte communication remain unclear. In this study, cumulus‐oocyte complexes (COCs) were subjected to HS at 42°C for 24 h during in vitro maturation. Cumulus expansion was impaired and oocyte quality was reduced with lower survival rate, polar body extrusion rate and early embryo developmental potentials. Oocytes and cumulus cells isolated from COCs demonstrated distinct responses to HS. In oocytes, mitochondrial DNA (mtDNA)‐encoded genes and heat shock protein (HSP)‐related genes were significantly down‐regulated, with significantly lower ATP content yet higher mitochondria reactive oxygen species (mROS) and caspase 3 activity. In contrast, CCs isolated from heat‐exposed COCs showed significant activation of mtDNAencoded genes and HSP‐related genes, which was accompanied by increased ATP content and caspase 3 activity yet unaltered mROS content. When denuded oocytes and isolated CCs were subjected to HS separately, ATP content in denuded oocytes decreased more extensively than that in oocytes isolated from heat‐exposed COCs. Also, mROS was significantly increased in CCs but not in denuded oocytes, which coincided with less pronounced increase of caspase 3 in denuded oocytes. Moreover, F‐actin, a marker of trans‐zonal projections, and occludin, a tight junction protein, and their co‐localization, were significantly reduced in heat‐exposed COCs. These results indicate that cumulus cells and oocytes show distinct HS responses, indicating disrupted cumulus‐oocytes communication in heat‐exposed porcine oocytes.

Support or Funding Information

This work was supported by the National Key Research and Development Program of China (2016YFD0500502), the National Basic Research Program of China (2014CB138502), the Priority Academic Program Development of Jiangsu Higher Education Institutions, and Jiangsu Collaborative Innovation Center of Meat Production and Processing, Quality and Safety Control.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Regional Adipose Tissue Immune Cell Profiles in Childhood‐Onset and Adult‐Onset Obesity
Authors: Murphy Jessica , Dam Vi , Delaney Kerri , Morais José A. , Tsoukas Michael , Santosa Sylvia
Published: June 2021

Background

We do not understand why individuals who have had obesity since childhood are at greater risk of metabolic disease.

Objective

To examine the effects of obesity onset and adipose tissue region (upper vs. lower body) on the proportion of adipose tissue immune cells.

Methods

We used flow cytometry to quantify the proportion of immune cells in the stromovascular fraction of abdominal (AB) and femoral (TH) subcutaneous adipose tissue from adults with childhood‐onset (n = 16) or adult‐onset (n = 22) obesity.

Results

The proportion of CD8+CD3+ T‐cells was greater in AB than TH in childhood‐onset obesity, while greater in TH than AB in adult‐onset obesity. There was no effect of obesity onset on other immune cell types. However, there was an overall effect of region where the proportion of CD45RA+CD8+CD3+ T‐cells was greater in TH than AB, and the proportion of CD206+CD68+ M2‐like macrophages was greater in AB than TH.

Conclusion

Our results show that there are regional differences in the adipose tissue immune cell profiles between childhood‐onset and adult‐onset obesity. It remains to be seen whether these differences are implicated in increasing the risk of metabolic disease.

Support or Funding Information

This research was funded by NSERC and the Heart and Stroke Foundation of Canada

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.