bluuebunny/crossref_metadata_2025_split huggingface dataset:
Kolaviron, A Flavonoid From Garcinia Kola Seeds, Ameliorates Experimental Colitis In Rats Exposed To Cadmium
Authors: Adegoke Adeoti Gbemisola , Olaleye Samuel Babafemi 
Published: June 2021

Cadmium has been shown to potentiate experimental ulcers in laboratory animals. Its ability to predispose humans to ulcerative colitis has also been postulated. Polyphenols and flavonoids found in plants possess antioxidant and metal binding potentials and may therefore, be beneficial in pathological conditions of oxidative stress resulting from exposure to heavy metals. In the present study, the effect of kolaviron, a biflavonoid complex from the seeds of Garcinia kola (Heckel) on colitis in cadmium‐exposed rats was investigated. Male Wistar rats (110–120 g) were randomly divided into five groups. Group 1 received distilled water, Group 2 received dimethyl sulphoxide, and served as negative and positive controls respectively. Group 3 received cadmium (10 mg/kg) while group 4 animals received Cadmium (10 mg/kg) in addition to kolaviron (200 mg/kg). The last group received kolaviron (200 mg/kg) alone. After four weeks of exposure to cadmium with or without kolaviron, colitis was induced in all animals in groups 2–5 by intrarectal administration of 2 mL of 4% Acetic acid (AA). Rats were sacrificed and colons resected at days 0, 3, 7 and 14 days of colitis induction. Assessment was based on symptoms of colitis such as weight loss and stool consistency using a diarrhoea score scale. Colon sample homogenates were analysed for concentrations of Malondialdehyde (MDA), Nitric oxide (NO), Glutathione (GSH), Sulfhydryl, activities of Superoxide dismutase (SOD), Na+‐K+ ATPase, Myeloperoxidase (MPO) by spectrophotometry and histology by hematoxylin and eosin staining. The weight loss (%) in Cadmium group by week 3 was (31.05 ± 0.04). Intrarectal administration of AA resulted in increased stool score in Cadmium group (0.80 ± 0.00) till day 14 post colitis induction. Macroscopic score was significantly (p < 0.05) increased in Cadmium group till day 14. MDA were significantly (p < 0.05) increased in Cadmium group at days 7 and 14. The notable increases in MPO and NO during AA‐induced colitis and cadmium exposure were significantly (p < 0.05) reduced by Kolaviron. Significant (p < 0.05) decreases in SOD, Na+‐K+ ATPase activities, sulfhydryl and GSH concentrations upon AA administration were attenuated in other groups except Cadmium group. Histology showed healing in all groups post colitis induction except in Cadmium group where histopathological damage was still evident at day 14. Kolaviron ameliorated the delayed healing of Acetic acid‐induced colitis during cadmium exposures in Wistar rats and may therefore, be useful as dietary intervention in cases of cadmium exposures.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Berberine hydrochloride induces brown adipogenesis
Authors: Li Lixin , Poudel Anil , Zhou Joseph 
Published: June 2021

Brown adipose tissue (BAT) dissipates energy in the form of heat through uncoupled respiration. Brown adipocytes originate from different precursor cells and can be induced by various stimuli. Pharmacologically induction of BAT thermogenesis has been shown to be a promising treatment for obesity‐related metabolic disease. In this study, we investigated the potential role of berberine hydrochloride (BBR), a naturally occurring plant alkaloid, in activating brown fat adipogenesis.3T3L1 preadipocyte cells, C2C12 myoblasts, and stromal vascular cells isolated from perigonadal fat of C57BL/6 mice were cultured in cell differentiation induction media for three days followed by differentiation maintenance media for another three days in the presence or absence of 10 μM of BBR. C2C12 differentiation media was also supplemented with PPARγ agonist rosiglitazone (0.5μM). Adipocyte differentiation was confirmed by oil red staining. Western blot analysis was performed on the differentiated cells to measure brown fat specific protein markers including UCP1, PCG‐1α, PPARγ, PRDM16 and CIDE‐A.BBR treatment significantly induced all of the above brown‐fat specific markers (p<0.005; n=4) in 3T3L1 cells, C2C12 myoblasts and stroma vascular blast cells. Specifically, UCP‐1 expression is increased 30.3 fold in 3T3L‐1, and 2.00 fold in primary adipocytes extracted from WAT respectively.Our results thus indicate that BBR induces brown adipogenesis, and is a promising treatment for obesity and obesity related metabolic diseases.Support or Funding InformationStart Up fund (Central Michigan University )This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Sox9‐independent Control of Intestinal Paneth Cell Differentiation and Function
Authors: Bankaitis Eric , Dajee Devesh , Magness Scott 
Published: June 2021

The genetic and signaling mechanisms controlling the generation and function of differentiated cell types are of continued interest. The intestinal Paneth cell (PC) is a post‐mitotic and dedicated secretory cell type with reported functions in supporting intestinal stem cell (ISC) maintenance in the crypt stem cell niche, and in secreting antimicrobial peptides to protect the crypt from potentially pathogenic luminal microbiota. Previous studies have reported a near‐complete loss of PCs (as defined by morphological and marker‐based criteria) in epithelium where the transcription factor Sox9 is genetically ablated, leading to the proposition that Sox9 is required for PC differentiation. Currently, however, the mechanisms through which Sox9 controls PC differentiation are not known. We have found evidence in Sox9 knockout (KO) small intestine (SI) that cells at the crypt base frequently express PC‐markers such as Lysozyme (Lyz) and Cryptdin, exhibit apical granules, and are post‐mitotic, all of which are features indicative of PC differentiation. The numbers of Lyz+ cells are highest in the distal Sox9 KO SI but are significantly reduced relative to control, and are essentially absent in the proximal SI, suggesting that regionally patterned signals may drive PC differentiation independent of Sox9. Apical granularity and expression levels of Lyz/Cryptdin are also altered, suggesting that while Sox9 KO ISCs can initiate PC differentiation, they are unable to reach full maturity. Antibody and qPCR based detection of PC gene products (including secreted antimicrobials, growth factors, and transcriptional regulators proposed to drive PC differentiation downstream of Sox9) are maintained in Sox9 KO cultured intestinal organoids and are elevated in organoids treated with pharmacological agents that drive the PC differentiation program. These results indicate that Sox9‐deficient ‘Paneth‐like’ cells are continually generated in culture and can be pushed toward PC differentiation with factors known to stimulate the PC differentiation program. We conclude that Sox9 is not required for the activation of at least sub‐sets of gene products associated with PC differentiation and function. Ongoing experiments are designed to investigate Sox9‐ dependent and independent pathways toward PC fate, and to characterize the functionality of Sox9−/− ‘Paneth‐like’ cells compared to controls.Support or Funding InformationRO1 DK091427‐05F32 DK115016‐01This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Relationship Between Sympathetic Baroreflex Sensitivity And Vascular Transduction
Authors: Taylor Chloe E , Hissen Sarah L , Brown Rachael , Macefield Vaughan G 
Published: June 2021

Sympathetic baroreflex sensitivity (BRS) is a tool used to quantify how effectively the baroreflex buffers beat‐to‐beat changes in blood pressure. However, current methods of assessment do not take into account vascular transduction, i.e. the response of the peripheral vasculature to vasoconstrictor drive. The aim of the study was to investigate the relationship between sympathetic BRS and vascular transduction. Muscle sympathetic nerve activity (MSNA, microneurography), blood pressure, heart rate and superficial femoral artery blood flow (Doppler ultrasound) were recorded at rest for 5 min in 28 healthy adults (18–31yrs, 13 males, 15 females). Sympathetic BRS was quantified by plotting MSNA burst incidence against mean diastolic pressure using 3 mmHg bins. For vascular transduction, the % change in leg vascular conductance was determined for 15 cardiac cycles following each MSNA burst and the nadir of the response was calculated. Linear regression analysis was used to examine the relationship between sympathetic BRS and vascular transduction in males and females separately. Sympathetic BRS was not significantly different between males (−2.9± 1.8) and females (−3.2 ± 1.6 %bursts/mmHg, p=0.68). However, vascular transduction was significantly lower in males (−5.5 ± 4.1) compared with females (9.1 ± 4.7 %change in ml/min/mmHg, p=0.04). In males there was a significant inverse relationship between sympathetic BRS and vascular transduction (r=0.78, p=0.002), but in females there was no significant relationship (r=0.34, p=0.21). The results suggest that in males those with high sympathetic BRS have low vascular transduction, and vice versa. This may represent a compensatory effect to ensure that blood pressure is regulated effectively. However, this relationship between sympathetic BRS and vascular transduction was not apparent in young females.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Evidence Supporting a Role for Autonomic Modulation of C‐Low Threshold Mechanoreceptors
Authors: Provost Makalele , Goolsby Bill , Hochman Shawn 
Published: June 2021

C‐low threshold mechanoreceptors (LTMRs) are a functionally distinct class of cutaneous sensory fibers believed to facilitate the sensing and coding of pleasurable touch. These afferents have also been implicated in neuropathic pain states including allodynia, in which normally innocuous tactile stimuli elicit painful sensations. The mechanisms that mediate changes in C‐LTMR function are incompletely understood. Autonomic activity has been implicated in the maintenance of various pain syndromes, yet increases in autonomic neural activity do not seem to influence the firing properties of cutaneous nociceptors. Autonomic modulation of cutaneous C‐LTMRs may contribute to allodynia and neuropathic pain, but this has not been studied. We developed an in vitro isolated truncal skin‐nerve preparation to study the recruitment properties of C‐LTMRs following optical stimulation of skin (TH::CHR2 mice) and the actions of modulatory transmitters when bath applied to the subcutaneous interior side of the skin. Extracellular spike recordings were obtained from dorsal cutaneous nerves. Optogenetic recruitment of C‐LTMRs was temperature sensitive with lower temperatures recruiting more units. Recruited C‐LTMRs had typical conduction velocities (ranging from 0.35 to 0.5 m/s) and an average spike half‐width of 0.53 ms (+/− 0.2ms). We assessed fatigability in response to repetitive optical stimuli at 10 Hz. Consistent with previous reports C‐LTMRs were highly fatigable, especially at higher temperatures. Application of acetylcholine resulted in increased spontaneous afferent activity, reduced C‐LTMR recruitment, and increased fatigability. Application of the cholinesterase inhibitor neostigmine also depressed C‐LTMR recruitment. Norepinephrine also decreased recruitment and increased fatigability but reduced the incidence of spontaneous activity. The norepinephrine transport inhibitor clomipramine also reduced C‐LTMR recruitment. Serotonin did not result in changes to recruitment or fatigability of C‐LTMRs, but increased spontaneous activity. However, the serotonin transport inhibitor citalopram did reduce C‐LTMR recruitment. Similarly, dopamine administration had no effect, but application of the dopamine transport inhibitor buproprion depressed C‐LTMR recruitment. We conclude that C‐LTMR activity can be modified by neuromodulatory transmission and studies are underway to further characterize this effect.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Ion Channel Activity in a 3‐hydroxyacyl‐coA dehydratase 1 (HACD1) Deficient Muscle Cell Line
Authors: Morgan Rhiannon Sarah , Walmsley Gemma , Dyer Steven , O'Brien Fiona , Staunton Caroline , Blondelle Jordan , Tiret Laurent , Piercy Richard , Barrett‐Jolley Richard 
Published: June 2021

Centronuclear myopathy in Labrador retrievers is the most common inherited neuromuscular disorder in dogs: this is caused by a mutation in a fatty acid processing gene, HACD1. The mutation is carried by 15–20% of the breed worldwide (Pele et al., 2005). These dogs present with generalised progressive weakness. A congenital myopathy due to a HACD1 mutation has also been documented in a human family, where infants have hypotonia and delayed motor milestones (Muhammed et al., 2013). The cellular mechanisms by which HACD1 mutation lead to muscle weakness are unknown, but since fatty acids interact with membrane ion channels we hypothesised that dysregulation of membrane function could be involved. To investigate this we have used a muscle cell line (C2C12) with shRNA knock down (KD) of HACD1 in combination with ion channel gene qPCR and patch‐clamp electrophysiology.To quantify changes in C2C12 myogenic differentiation following HACD1 KD, we used a fusion index calculation on day 8 myotubes stained for the differentiation marker MF20 and counted nuclei that were present in MF20 positive myotubes against those that were in undifferentiated, MF20 negative cells. No significant difference in myotube fusion was observed in HACD1‐KD cells compared to control (n=2).Before beginning patch‐clamp studies on C2C12 cells we used qPCR to identify potential ion channel targets in control C2C12 myoblasts. We found expression of (<30ct) of 21 ion channels, including those of particular interest to calcium homeostasis such as the voltage‐gated Ca2+ channels; Cacna1c, cacna1g, the non‐selective cation (NSC) channels; TRPV4, TRPV2, TRPC1 and a Ca2+‐activated K+ channel; Kcnn3.To functionally identify channel gating we used cell‐attached patch‐clamp electrophysiology of control and HACD1‐KD C2C12 myoblasts. K‐means clustering revealed five statistically significant clusters of ion channel activity in these cells (p<0.05). Assuming an RMP of −15mV (Tanaka et al., 2017) clusters had centeroids of;(i) conductance 21 ± 0.4pS, Vrev 30 ± 6mV (ii) 28 ± 6pS, −38 ± 4mV (iii) 29 ± 0.7pS, −78 ± 4mV (iv) 40 ± 6pS, 5 ± 3mV (v) 94 ± 13, 16 ± 44mV.In conclusion, the phenotype of the smallest ion channel conductance (28pS) would be consistent with the Ca2+‐activated K+ channel KCNN3, and the 94pS consistent with any of the three TRP channels identified by qPCR. These will be further characterised with pharmacological inhibitors and in the future we will examine if the populations change as the control and HACD1‐KD myoblasts differentiate into myotubes.Support or Funding InformationAuthors would like to thank J Blondelle & L Tiret from the Alfort Veterinary School for the cells used in these experiments.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 Effects of Leucine Supplementation on Skeletal Muscle Mitochondrial Content and Protein Turnover in Tumor Bearing Mice
Authors: Lee Harold , Persinger Aaron , Lee Kevin , Jones Michelle , Puppa Melissa 
Published: June 2021

IntroductionMany forms of cancer are associated with loss of lean body mass, a significant indicator of increased mortality. This effect is commonly attributed to decreased protein synthesis and stimulation of proteolytic pathways within the skeletal muscle due to metabolic disturbances from the tumor burden. The branched chain amino acid (BCAA) leucine has been shown to improve protein synthesis, insulin signaling, and mitochondrial biogenesis, key signaling pathways influenced by tumor signaling.PurposeOur study aimed to elucidate the effects of leucine supplementation on mitochondrial biogenesis, within the Lewis Lung Carcinoma (LLC) implantation model. We hypothesized that LLC implantation will impair mitochondrial biogenesis and protein synthesis leading to a loss of muscle mass, with leucine attenuating these effects.MethodsTwenty male C57BL/6 mice were divided into four equal groups (n=5) of equivalent body weight: Chow, leucine (Leu), LLC, LLC+Leu. At the age of 9–10 weeks, mice received a subcutaneous injection of 1×106 LLC cells or phosphate buffered saline (PBS). Leu groups were then switched to diet supplemented with 5% leucine (w/w). Upon euthanasia, muscle and tumors were weighed and collected. Measures of protein synthesis, mitochondrial biogenesis, and inflammation in the gastrocnemius muscle were assessed via western blot analysis.ResultsWhile body mass was not different between groups, gastrocnemius mass was decreased in the LLC+Leu group relative to the LLC group (p=0.040). Protein synthesis, quantified through the SUnSET technique, was decreased in LLC mice (p=0.001). Phosphorylation of STAT3, an indicator of inflammation was decreased in the Leu group relative to the control (p=0.019), but did not significantly attenuate the inflammatory effect of LLC implantation (p=0.619). Peroxisome proliferator‐activated receptor Gamma Co‐activator 1‐α (PGC‐1α), a marker of mitochondrial biogenesis, was increased in LLC+Leu relative to LLC (p=0.001). Finally, LLC implantation decreased mitochondrial content as measured by Cytochrome C (p=0.015), mitochondrial complexes III (p=0.006) and V (p=0.041).ConclusionWithin our study, leucine supplementation was unable to preserve protein synthesis or mitochondrial content associated with LLC implantation. However, Leucine supplementation was able to increase mitochondrial biogenesis signaling.Support or Funding InformationUniversity of Memphis School of Health Studies Faculty GrantThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Comparative Pharmacokinetic Profile of Three Batches of Ovine Enoxaparin and One Batch of Branded Enoxaparin
Authors: Rangnekar Varun , Jeske Walter , Yao Yiming , Kouta Ahmed , Hoppensteadt Debra , Duff Rick , Fareed Jawed 
Published: June 2021

BackgroundHeparins can be derived from tissues of a variety of different species. Although current pharmaceutical grade heparin is derived almost exclusively from porcine intestinal mucosa, there is interest within the FDA for diversifying the sourcing of heparin in the US to protect against supply shortages and economically motivated adulteration as was seen in 2008. Because sheep‐derived (ovine) heparin is structurally similar to porcine heparin, it is expected that low molecular weight heparin (LMWH) derived from ovine‐heparin will be comparable to porcine‐derived LMWH. This study will compared the pharmacokinetic behavior of ovine LMWH with that of enoxaparin in non‐human primates.Materials and MethodsEnoxaparin (Lovenox, Sanofi‐Aventis, Bridgewater, NJ) and three batches of ovine LMWH produced by benzylaztion/alkaline hydrolysis (Suzhou Ronnsi Pharma, Suzhou, China) were used in this study. Following anesthesia, a baseline blood sample was collected from non‐human primates (Macaca mulatta; n=4/LMWH). LMWH was administered subcutaneously (SC) at a dose of 1 mg/kg. Additional blood samples were collected at 2, 4 and 6 hours post‐administration. Circulating drug concentrations determined using anti‐Xa and anti‐thrombin assays were used to calculate pharmacokinetic parameters such as half‐life, area under the curve, systemic clearance and volume of distribution using the PKSolver add‐in for Microsoft Excel. Tissue factor pathway inhibitor (TFPI) levels were measured by ELISA.ResultsIn vitro studies demonstrated that the ovine LMWHs met pharmacopoeial requirements for enoxaparin in terms of potency and molecular weight distribution. In the samples collected at 2, 4, and 6 hours, comparable levels of branded enoxaparin and the three batches of ovine enoxaparin were observed using both anti‐Xa and anti‐thrombin assays, with the concentration vs. time curves being nearly superimposable. Consistent with the similarity observed with the plasma drug concentrations, no significant differences were observed between pharmacokinetic parameters calculated for branded enoxaparin and ovine LMWH. TFPI levels returned to baseline levels by 6 hours in ovine LMWH‐treated animals, but remained slightly elevated in animals treated with branded enoxaparin.DiscussionAt a 1 mg/kg SC dosage, three ovine enoxaparin preparations produced comparable anti‐Xa and anti‐IIa effects which were similar to the ones observed with branded enoxaparin. The PK parameters calculated based on this data were also comparable for both groups. It is concluded that the SC pharmacokinetics of the three batches of ovine enoxaparin were not only comparable between batched, but were also similar to the branded product.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.