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1.
Phys Occup Ther Pediatr ; : 1-18, 2024 Jun 07.
Article in English | MEDLINE | ID: mdl-38845534

ABSTRACT

AIM: Trauma-informed care (TIC) is a strengths-based organizational approach which recognizes the prevalence and impact of trauma on an individual's neurological, sensorimotor, and emotional responses and development. There is limited research on pediatric occupational therapy providers' (OTPs) and physical therapy providers' (PTPs) perceptions on incorporating TIC into their practice. This pilot study aims to describe the confidence, attitudes, barriers, and education of pediatric OTPs and PTPs regarding TIC. METHOD: OTPs and PTPs (n = 87) working in pediatric settings in the U.S. completed an electronic survey regarding their perceptions of TIC. RESULTS: OTPs reported higher confidence in TIC compared to PTPs. Most participants reported favorable attitudes toward TIC, yet no statistical significance was identified between the two professions' overall attitudes toward TIC. Collectively, participants reported lack of time and training as barriers. There was a statistically significant difference between how the two professions rated competency, training, and concern over clients' retraumatization. There was significant association between years of professional experience and TIC education. CONCLUSION: Preliminary findings indicate an overall agreement of OTPs and PTPs with principles of TIC and common barriers to TIC practice. There are implications to further integrate opportunities for TIC training into foundational education and professional practice.

2.
Personal Disord ; 15(1): 60-73, 2024 Jan.
Article in English | MEDLINE | ID: mdl-38206863

ABSTRACT

Borderline personality disorder (BPD) is a serious and understudied mental health condition associated with profound personal and public health consequences. Methodological differences in characterizing BPD may limit understanding the scope of the disorder's prevalence and effect. For example, using different diagnostic rules for BPD can affect apparent prevalence, comorbidity, and clinical presentation. This study examined how differences in diagnostic rules used to assign BPD diagnosis impacted its prevalence and associations with clinically relevant variables (e.g., demographics, comorbidity, treatment-seeking). Participants were a nationally representative sample of 36,309 noninstitutionalized U.S. adults. All variables were assessed via clinical interview (Alcohol Use Disorder and Associated Disabilities Interview Schedule-5). Six diagnostic rules determined BPD status. We used frequencies to examine prevalence rates of and associations between BPD and other clinical variables, and logistic regressions to examine the associations between each BPD variable and the other outcomes. The prevalence of BPD ranged widely-from 0.5% to 11.4%-per the diagnostic rule used. Associations between BPD diagnosis and various outcomes and clinical variables generally remained stable across all diagnostic rules, though effects became more extreme as diagnostic rules became more restrictive. Additionally, meaningful differences emerged as a function of the number of items used (30 vs. 18 items) even with no other changes to diagnostic rules. The field examining BPD and associated problem behaviors should critically consider how to most effectively characterize BPD to understand these problems more accurately and optimize the generalizability of findings. (PsycInfo Database Record (c) 2024 APA, all rights reserved).


Subject(s)
Alcoholism , Borderline Personality Disorder , Adult , Humans , Borderline Personality Disorder/diagnosis , Borderline Personality Disorder/epidemiology , Prevalence , Comorbidity , Alcohol Drinking
4.
Cell Immunol ; 391-392: 104751, 2023.
Article in English | MEDLINE | ID: mdl-37480823

ABSTRACT

Mast cells initiate allergic inflammatory immune responses and play a role in disease by releasing various inflammatory and immunomodulatory mediators. Several mast cell-lines and primary cultured cells have been used as mast cell models with inconsistent results among research groups. Bone marrow-derived mast cells (BMMC) cultured from mouse bone marrow progenitor cells are often used as a representative model of mucosal mast cell behaviour, however their reported phenotype is variable due to inconsistent culture protocols. RBL-2H3 is a rat basophilic histamine-releasing cell line that has some characteristics of both mast cells and basophils but is not a true representation of either cell type. The murine mast cell line MC/9 is an IL-3-dependent mucosal mast cell model but has limited mast cell characteristics. In this study, we have compared the response of BMMC (derived from C57BL/6 male or female mice), two sources of RBL-2H3 (purchased directly from ATCC and a lab curated culture), and MC/9 (ATCC) at several critical stages to some common stimuli (IgE/Ag, A23187) and analyzed mast cell morphology, expression level of common mast cell surface markers (CD117 and FcεRI), protease expression, and function (growth kinetics, viability, ROS production, degranulation, cytokine release and FcεRI signaling). The objective of this study was to provide insight into the effects of culture conditions, biological sex, and age of the cells on variability among reported phenotypes and, to determine optimal conditions for activation of these cells. Our data show that factors that are often overlooked such as source, age and biological sex of mast cells play an integral role in phenotypic outcomes and may account for the reported variability in their function.


Subject(s)
Basophils , Mast Cells , Rats , Mice , Male , Female , Animals , Rodentia , Mice, Inbred C57BL , Cell Line , Cell Degranulation
5.
Front Pharmacol ; 14: 1102465, 2023.
Article in English | MEDLINE | ID: mdl-37124206

ABSTRACT

Recent interest in mushrooms and their components as potential therapies for mental health, along with recent government and health authority approvals, has necessitated a more comprehensive understanding of their effects on the cellular microenvironment of the brain. Amanita muscaria has been ingested as a treatment for a variety of ailments for centuries, most notably those affecting the central nervous system and conditions associated with neuroinflammation. However, the effects of these extracts on neuroinflammatory cells, such as microglia, are unknown. The effect of commercially-sourced A. muscaria extract (AME-1) on human microglial cell line (HMC3) expression of surface receptors such as CD86, CXCR4, CD45, CD125 and TLR4 was determined by flow cytometry. AME-1 upregulated expression of all of these receptors. The effect of AME-1 on HMC3 production of IL-8 and IL-6 was determined and compared to tumor necrosis factor (TNF), polyinosinic-polycytidylic acid [poly(I:C)], substance P and lipopolysaccharide (LPS), all known activators of HMC-3 and primary microglia. HMC3 produced both IL-8 and IL-6 when activated with LPS, TNF and poly(I:C) but not when they were activated with substance P. Although AME-1 at higher concentrations increased IL-8 production of HMC3 on its own, AME-1 notably potentiated HMC3 production of IL-8 in response to poly(I:C). AME-1 altered expression of toll-like receptor 3 (TLR3) mRNA but not surface protein by HMC3. AME-1 also did not significantly alter expression of retinoic acid-inducible gene I (RIG-I) or melanoma differentiation-associated protein 5 (MDA5), both cytosolic sensors of dsRNA. Metabolomics analysis showed that AME-1 contained several metabolites, including the autophagy inducer, trehalose. Like AME-1, trehalose also potentiated HMC3 poly(I:C) mediated production of IL-8. This study suggests that A. muscaria extracts can modify HMC3 inflammatory responses, possibly due to their trehalose content.

6.
J Equine Vet Sci ; 127: 104828, 2023 08.
Article in English | MEDLINE | ID: mdl-37245655

ABSTRACT

Calming supplements are common in the equine industry. This study tested the hypothesis that Phytozen EQ, a blend of citrus botanical oils, magnesium, and yeast would reduce startle response as well as reduce behavioral and physiological signs of stress in young (1.5-6 years of age) horses (n = 14) when tied in isolation and when trailered in isolation. During the 59-day trial, horses were assigned to either the control (CON; n = 7) or treatment (PZEN; n = 7) group that received 56 g of Phytozen EQ daily. Horses underwent a 10-minute isolation test on d 30 and a 15 minute individual trailering test on day 52 or 55. For both tests, blood samples were obtained pre, immediately after, and 1-hour post for analysis of plasma cortisol concentrations, which were analyzed by repeated measures ANOVA. On day 59, horses underwent a startle test, for which time to travel 3 m and total distance traveled were recorded. These data were analyzed using a T-test. During trailering, PZEN horses tended to have lower overall geometric mean (lower, upper 95% confidence interval) cortisol concentrations than CON (81 [67, 98] vs. 61 [48, 78] ng/mL; P = .071). For the startle test, PZEN horses tended to have longer geometric mean times to travel 3 m than CON horses (1.35 [0.39, 4.70] vs. 0.26 [0.07, 0.91 seconds, P = 0.064). Other data points were not different between treatments (P > .1). It is possible that this dietary supplement could have beneficial calming effects on horses undergoing trailering or in novel situations.


Subject(s)
Dietary Supplements , Hydrocortisone , Horses , Animals , Dietary Supplements/analysis
7.
Front Bioeng Biotechnol ; 11: 1160460, 2023.
Article in English | MEDLINE | ID: mdl-37113661

ABSTRACT

Introduction: Mast cells are highly granulated tissue-resident leukocytes that require a three-dimensional matrix to differentiate and mediate immune responses. However, almost all cultured mast cells rely on two-dimensional suspension or adherent cell culture systems, which do not adequately reflect the complex structure that these cells require for optimal function. Methods: Crystalline nanocellulose (CNC), consisting of rod-like crystals 4-15 nm in diameter and 0.2-1 µm in length, were dispersed in an agarose matrix (12.5% w/v), and bone marrow derived mouse mast cells (BMMC) were cultured on the agarose/CNC composite. BMMC were activated with the calcium ionophore A23187 or immunoglobulin E (IgE) and antigen (Ag) to crosslink high affinity IgE receptors (FcεRI). Results: BMMC cultured on a CNC/agarose matrix remained viable and metabolically active as measured by reduction of sodium 3'-[1-[(phenylamino)-carbony]-3,4-tetrazolium]-bis(4-methoxy-6-nitro) benzene-sulfonic acid hydrate (XTT), and the cells maintained their membrane integrity as analyzed by measuring the release of lactate dehydrogenase (LDH) and propidium iodide exclusion by flow cytometry. Culture on CNC/agarose matrix had no effect on BMMC degranulation in response to IgE/Ag or A23187. However, culture of BMMC on a CNC/agarose matrix inhibited A23187-and IgE/Ag-activated production of tumor necrosis factor (TNF) and other mediators such as IL-1ß, IL-4, IL-6, IL-13, MCP-1/CCL2, MMP-9 and RANTES by as much as 95%. RNAseq analysis indicated that BMMC expressed a unique and balanced transcriptome when cultured on CNC/agarose. Discussion: These data demonstrate that culture of BMMCs on a CNC/agarose matrix promotes cell integrity, maintains expression of surface biomarkers such as FcεRI and KIT and preserves the ability of BMMC to release pre-stored mediators in response to IgE/Ag and A23187. However, culture of BMMC on CNC/agarose matrix inhibits BMMC production of de novo synthesized mediators, suggesting that CNC may be altering specific phenotypic characteristics of these cells that are associated with late phase inflammatory responses.

8.
Molecules ; 27(19)2022 Oct 08.
Article in English | MEDLINE | ID: mdl-36235240

ABSTRACT

Mast cells (MC) synthesize and store proinflammatory mediators and are centrally important in atopic diseases such as asthma and atopic dermatitis. Quercetin a and resveratrol are plant derived polyphenolic compounds with anti-inflammatory properties that inhibit MC degranulation and mediator release. However, the underlying mechanism of these inhibitory effects on MC is poorly understood and it is unclear whether this is a general effect on all MC phenotypes. We have characterized and compared the effects of quercetin with resveratrol on human (LAD2) and mouse (MC/9 and BMMC) MC mediator release, receptor expression and FcεRI signaling to better understand the mechanisms involved in quercetin and resveratrol-mediated inhibition of MC activation. Quercetin significantly decreased the expression of FcεRI by BMMC and MC/9, although the effects on MC/9 were associated with a significant reduction in cell viability. Quercetin also inhibited antigen-stimulated TNF release by BMMC. Although neither quercetin nor resveratrol significantly altered antigen-stimulated BMMC degranulation or downstream signaling events such as phosphorylation of spleen tyrosine kinase (SYK) or extracellular signal-regulated kinase 1/2 (ERK), resveratrol inhibited ERK phosphorylation and FcεRI- stimulated degranulation in LAD2. Our data suggests that quercetin and resveratrol inhibit human and mouse MC differentially and that these effects are associated with modification of FcεRI expression, signaling (phosphorylation of SYK and ERK) and mediator release.


Subject(s)
Mast Cells , Receptors, IgE , Animals , Antigens/metabolism , Cell Degranulation , Congenital Disorders of Glycosylation , Humans , Mast Cells/metabolism , Mice , Mitogen-Activated Protein Kinase 3/metabolism , Quercetin/metabolism , Quercetin/pharmacology , Receptors, IgE/metabolism , Resveratrol/metabolism , Resveratrol/pharmacology , Syk Kinase/metabolism
9.
J Anim Sci ; 100(8)2022 Aug 01.
Article in English | MEDLINE | ID: mdl-35908781

ABSTRACT

Optimal athletic performance requires meeting the energetic demands of the muscle fibers, which are a function of myosin ATPase enzymatic activity. Skeletal muscle with a predominant oxidative metabolism underlies equine athletic success. Sodium butyrate, a short-chain fatty acid, can affect muscle fiber composition in pigs. To determine if a similar scenario exists in horses, 12 adult Thoroughbred geldings (7.4 ± 0.6 yr of age; mean ± SEM) were fed 16 g of calcium butyrate (CB) or an equivalent amount of carrier (CON) daily for 30 d in a crossover design. Middle gluteal muscle biopsies were collected before and after the feeding trial for immunohistochemical determination of fiber type, and RNA and protein isolation. After 30 d, CB increased (P < 0.05) the percentage of type IIA fibers and tended (P = 0.13) to reduce the numbers of type IIX fibers in comparison to control (CON). No changes (P > 0.05) in type I, IIA, or IIX fiber size were observed in response to CB. No differences (P > 0.05) were noted in the abundance of succinate dehydrogenase (SDH) protein or activity between horses receiving CB or CON. Myogenin mRNA abundance was unaffected (P > 0.05) by 30 d of CB supplementation. The increase in type IIA fibers in the absence of altered mitochondrial SDH enzymatic activity suggests that CB affects myosin ATPase expression independent of altered metabolism.


The largest tissue in the body, skeletal muscle, is a heterogeneous mix of fibers that are categorized based on their primary source of energy production and speed of contraction. Evidence suggests that Thoroughbred horses with a greater percentage of type IIA, fast-twitch, oxidative fibers are more successful than those with fewer. Pigs fed a diet supplemented with butyrate contained a greater percentage of oxidative muscle fibers. This study examined the ability of calcium butyrate (CB), a short-chain fatty acid, to alter muscle fiber composition in horses. Adult Thoroughbred geldings were supplemented with a placebo or CB for 30 d, and gluteus medius muscle biopsies were retrieved and analyzed for fiber type, myogenin expression, and succinate dehydrogenase (SDH) activity. Results demonstrate a small increase in the percentage of type IIA fibers without a change in SDH activity, a marker of oxidative metabolism. Myogenin expression remained unaffected by CB supplementation. These efforts underscore the need for further research to validate improved exercise performance in response to CB supplementation and identify a mechanism of action for the fatty acid in the equine skeletal muscle.


Subject(s)
Calcium , Myosin Heavy Chains , Animals , Butyrates/metabolism , Calcium/metabolism , Dietary Supplements , Horses , Male , Muscle Fibers, Skeletal/metabolism , Muscle, Skeletal/metabolism , Myosin Heavy Chains/genetics , Myosin Heavy Chains/metabolism , Myosins , Oxidative Stress , Swine
10.
Gels ; 9(1)2022 Dec 22.
Article in English | MEDLINE | ID: mdl-36661773

ABSTRACT

Mono-N-alkylated primary oxalamide derivatives with different sized branched alkyl tail-groups were excellent low molecular weight gelators for a variety of different organic solvents with different polarities and hydrogen-bonding abilities. Solvent-gelator interactions were analyzed using Hansen solubility parameters, while 1H NMR and FTIR spectroscopy were used to probe the driving forces for the supramolecular gelation. The molecular structures of the twin tail-groups did not significantly affect the supramolecular gelation behavior in different solvents. However, for select solvents, the molecular structures of the tail-groups did have a significant effect on gel properties such as the critical gelator concentration, thermal stability, gel stiffness, gel strength, network morphology, and molecular packing. Finally, metabolic activity studies showed that the primary alkyl oxalamide gelators had no effect on the metabolic activity of mouse immune cells, which suggests that the compounds are not cytotoxic and are suitable for use in biomedical applications.

11.
J Vis Exp ; (171)2021 05 11.
Article in English | MEDLINE | ID: mdl-34057448

ABSTRACT

Three-dimensional (3D) bioprinting utilizes hydrogel-based composites (or biomaterial inks) that are deposited in a pattern, forming a substrate onto which cells are deposited. Because many biomaterial inks can be potentially cytotoxic to primary cells, it is necessary to determine the biocompatibility of these hydrogel composites prior to their utilization in costly 3D tissue engineering processes. Some 3D culture methods, including bioprinting, require that cells be embedded into a 3D matrix, making it difficult to extract and analyze the cells for changes in viability and biomarker expression without eliciting mechanical damage. This protocol describes as proof of concept, a method to assess the biocompatibility of a crystalline nanocellulose (CNC) embedded agarose composite, fabricated into a 24-well culture system, with mouse bone marrow-derived mast cells (BMMCs) using flow cytometric assays for cell viability and biomarker expression. After 18 h of exposure to the CNC/agarose/D-mannitol matrix, BMMC viability was unaltered as measured by propidium iodide (PI) permeability. However, BMMCs cultured on the CNC/agarose/D-mannitol substrate appeared to slightly increase their expression of the high-affinity IgE receptor (FcεRI) and the stem cell factor receptor (Kit; CD117), although this does not appear to be dependent on the amount of CNC in the bioink composite. The viability of BMMCs was also assessed following a time course exposure to hydrogel scaffolds that were fabricated from a commercial biomaterial ink composed of fibrillar nanocellulose (FNC) and sodium alginate using a 3D extrusion bioprinter. Over a period of 6-48 h, the FNC/alginate substrates did not adversely affect the viability of the BMMCs as determined by flow cytometry and microtiter assays (XTT and lactate dehydrogenase). This protocol describes an efficient method to rapidly screen the biochemical compatibility of candidate biomaterial inks for their utility as 3D scaffolds for post-print seeding with mast cells.


Subject(s)
Bioprinting , Cell Culture Techniques , Ink , Animals , Biocompatible Materials , Bone Marrow , Mice , Printing, Three-Dimensional , Sepharose , Tissue Engineering , Tissue Scaffolds
12.
J Equine Vet Sci ; 101: 103455, 2021 06.
Article in English | MEDLINE | ID: mdl-33993938

ABSTRACT

Exercising horses are commonly plagued by muscle fatigue and soreness, which can result in reduced performance ability. In the present study, ten unconditioned horses were fed 200g per day DigestaWell NRG, a commercial dietary supplement containing Yucca schidigera and Trigonella foenum-graecum, two herbs shown in other species to reduce post-exercise muscle pain and soreness. A control, unsupplemented group contained ten horses of similar age, breed, and gender. Horses completed a 50 minutes, ridden standardized exercise test of moderate intensity immediately prior to (Period1) and after 28 days of supplementation (Period2). Muscle soreness and tightness were evaluated 24 hours prior to and after each exercise test and used to determine the percent increase in post-exercise muscle soreness and tightness. Blood samples were collected before, and at 10 and 30 minutes, and 1, 4, and 24 hours post exercise. Plasma was analyzed for glucose, lactate, non-esterified fatty acid, tumor necrosis factor-α, and interleukin-1ß concentrations. Data were analyzed by repeated measures ANOVA using SAS Enterprise Guide v. 7.1. No changes in plasma parameters were indicated between periods for unsupplemented horses (P > 0.1) during Period2, excepting glucose, which was greater during Period2 (P = 0.018). Supplemented horses had lesser concentrations of tumor necrosis factor-α (P = 0.016) and lactate (P = 0.058) during Period2 than during Period1. During Period2, supplemented horses experienced a smaller percent increase in post exercise muscle soreness (P = 0.031). DigestaWell NRG supplementation may benefit unconditioned horses undergoing moderate intensity exercise through reducing lactate production and inflammation.


Subject(s)
Horse Diseases , Physical Conditioning, Animal , Animals , Dietary Supplements , Horses , Myalgia/drug therapy , Myalgia/veterinary , Pilot Projects , Plant Breeding
13.
J Equine Vet Sci ; 88: 102938, 2020 05.
Article in English | MEDLINE | ID: mdl-32303306

ABSTRACT

Insulin resistance (IR) is characterized by an increase in biomarkers of systemic inflammation and susceptibility to laminitis in horses. Impacts on reproduction include a lengthened interovulatory period in horses. Dietary omega-3 (docosahexaenoic acid [DHA]) promotes anti-inflammatory processes, has been implicated in health benefits, and can reduce cytokine secretion. This preliminary study investigated the impact of IR as well as the influence of dietary supplementation (DHA) on the uterine fluid proteome in early pregnant horses. Mares were artificially inseminated; uterine fluid and embryos were collected on d 12.5 after ovulation. Uterine fluid was pooled for metabolic and diet categories (n = 8; n = 2 per metabolic and dietary status) and concentrated, and the proteome was analyzed using tandem mass spectrometry (iTRAQ). Five proteins met differential abundance criteria (±1.5-fold change, P < .05) in all comparisons (Control C, IS vs. C, IR; C, IS vs. DHA, IS; C, IR vs. DHA, IR). Serum amyloid A, afamin, and serotransferrin were upregulated in C, IR mares but downregulated in DHA, IR mares when compared to C, IS and C, IR, respectively. Quantitative PCR supported mass spectrometry results. The presence of serum amyloid A and serotransferrin in histotroph of IR mares potentially indicates an inflammatory response not seen in IS counterparts. These preliminary findings provide novel evidence on the potential impact of insulin resistance and DHA supplementation on the secreted equine uterine proteome during early pregnancy.


Subject(s)
Fatty Acids, Omega-3 , Proteome , Animals , Diet/veterinary , Docosahexaenoic Acids , Female , Horses , Pregnancy , Tandem Mass Spectrometry/veterinary
14.
Animals (Basel) ; 10(3)2020 Mar 13.
Article in English | MEDLINE | ID: mdl-32183167

ABSTRACT

There is little information available to describe the inflammatory consequences of and recovery from moderate-intensity exercise bouts in hunting dogs. The purpose of the current study is to generate pilot data on the appearance and disappearance of biomarkers of inflammation and inflammation resolution following a typical one-hour exercise bout in basset hounds. Four hounds were set out to find a scent and freely adopted running or walking over wooded terrain for approximately one hour. Venous blood samples were obtained before the exercise and at 1, 2, 4, 6, and 10 hours following cessation of exercise and were analyzed for biomarkers of inflammation (prostaglandin E2 (PGE2), nitric oxide (NO), interleukin 1ß (IL-1ß)) tumour necrosis factor-α (TNF-α)), and inflammation resolution (resolvin D1 (RvD1)). There was an increase in inflammation one hour after the exercise, shown by a significant increase in PGE2. Following this peak, PGE2 steadily declined at the same time as RvD1 increased, with RvD1 peaking at six hours. This pilot study provides evidence that dogs that undergo an hour of hunt exercise experience transient inflammation that peaks one hour after the end of exercise; inflammation resolution peaks six hours after the end of exercise. Future studies should seek to further understand the distinct and combined roles of PGE2 and RvD1 in dog adaptation to exercise stress.

15.
Article in English | MEDLINE | ID: mdl-32098057

ABSTRACT

Swachh Bharat Abhiyan, India's flagship sanitation intervention, set out to end open defecation by October 2019. While the program improved toilet coverage nationally, large regional disparities in construction and use remain. Our study used ethnographic methods to explore perspectives on open defecation and latrine use, and the socio-economic and political reasons for these perspectives, in rural Bihar. We draw on insights from social epidemiology and political ecology to explore the structural determinants of latrine ownership and use. Though researchers have often pointed to rural residents' preference for open defecation, we found that people were aware of its many risks. We also found that (i) while sanitation research and "behavior change" campaigns often conflate the reluctance to adopt latrines with a preference for open defecation, this is an erroneous conflation; (ii) a subsidy can help (some) households to construct latrines but the amount of the subsidy and the manner of its disbursement are key to its usefulness; and (iii) widespread resentment towards what many rural residents view as a development bias against rural areas reinforces distrust towards the government overall and its Swachh Bharat Abhiyan-funded latrines in particular. These social-structural explanations for the slow uptake of sanitation in rural Bihar (and potentially elsewhere) deserve more attention in sanitation research and promotion efforts.


Subject(s)
Defecation , Toilet Facilities , Adolescent , Adult , Aged , Anger , Female , Humans , India , Male , Middle Aged , Rural Population , Sanitation , Social Responsibility , Young Adult
16.
J Anim Physiol Anim Nutr (Berl) ; 103(1): 283-294, 2019 Jan.
Article in English | MEDLINE | ID: mdl-30284316

ABSTRACT

The objective of this study was to examine markers of whole-body and muscle protein metabolism in aged horses fed a diet typical for North American aged horses, supplemented with amino acids. In a replicated Latin square design, six aged horses (20 ± 1.1 years) were studied while receiving each of three isocaloric, isonitrogenous diets, a control treatment concentrate (CON; 100 mg/kg-1 BW day-1 lysine, 84 mg kg-1  day-1 threonine, 51 mg kg-1  day-1 methionine), LYS/THR (134 mg kg-1 BW day-1 lysine, 110 mg kg-1 BW day-1 threonine, 52 mg kg-1 BW day-1 methionine) and LYS/THR/MET (132 mg kg-1 BW day-1 lysine, 112 mg kg-1 BW day-1 threonine, 62 mg kg-1 BW day-1 methionine). In each 15-days period, urine and faeces were collected for assessment of nitrogen balance. Blood samples were collected before and after feeding for analysis of plasma urea nitrogen (PUN), glucose, insulin and plasma amino acid concentrations. Skeletal muscle samples were collected for measurement of proteins associated with muscle protein synthesis and degradation, and horses underwent stable isotope infusion procedures for comparison of differences in whole-body rates of protein synthesis and degradation. There was no effect of treatment on relative abundance of proteins involved in protein synthesis, nitrogen retention or phenylalanine kinetics. PUN concentrations tended to be higher for LYS/THR (p = 0.054) and were higher for LYS/THR/MET (p = 0.0056) than for CON. Atrogin-1 abundance tended to be higher in the post-absorptive state for the CON treatment (p = 0.07), indicating that amino acid supplementation resulted in less muscle protein degradation when horses were in the post-absorptive state. However, lack of differences in nitrogen retention and phenylalanine kinetics indicated that whole-body protein metabolism was not improved, and higher PUN concentrations in the supplemented diets suggest that the supplemented amino acids may have been catabolized. Amino acid availability was not limiting protein synthesis in the sedentary aged horses in this study when fed the CON diet.


Subject(s)
Aging , Amino Acids/administration & dosage , Horses/physiology , Muscle Proteins/metabolism , Animal Feed/analysis , Animal Nutritional Physiological Phenomena , Animals , Blood Glucose , Blood Urea Nitrogen , Connectin/drug effects , Connectin/metabolism , Cross-Over Studies , Diet/veterinary , Dietary Supplements , Gene Expression Regulation/drug effects , Insulin/blood , Muscle Proteins/genetics , Random Allocation
17.
Toxicology ; 404-405: 25-32, 2018 07 01.
Article in English | MEDLINE | ID: mdl-29729928

ABSTRACT

To examine the functional role of peroxisome proliferator-activated receptor-ß/δ (PPARß/δ) and PPARγ in skin cancer, stable cell lines were created in the A431 human squamous cell carcinoma cell line. Expression of PPAR target genes was greatly enhanced in response to ligand activation of PPARß/δ or PPARγ in A431 cells expressing these receptors. PPARß/δ expression blocked the cell cycle at the G2/M phase, and this effect was increased by ligand activation. Ligand activation of PPARß/δ markedly inhibited clonogenicity as compared to vehicle-treated controls. Similarly, ligand activation of PPARγ in A431 cells expressing PPARγ resulted in reduced clonogenicity. Expression of either PPARß/δ or PPARγ markedly reduced tumor volume in ectopic xenografts, while ligand activation of these receptors had little further influence on tumor volume. Collectively, these studies demonstrate that stable expression and activation of PPARß/δ or PPARγ in A431 cells led to reduced tumorigenicity. Importantly, PPAR expression or ligand activation had major impacts on clonogenicity and/or tumor volume. Thus, PPARß/δ or PPARγ could be therapeutically targeted for the treatment of squamous cell carcinomas.


Subject(s)
Carcinogenesis/metabolism , Carcinoma, Squamous Cell/metabolism , Cell Cycle/physiology , PPAR delta/biosynthesis , PPAR-beta/biosynthesis , Skin Neoplasms/metabolism , Animals , Carcinoma, Squamous Cell/prevention & control , Cell Cycle Checkpoints/physiology , Cell Line, Tumor , Female , Humans , Mice, Nude , Skin Neoplasms/prevention & control , Xenograft Model Antitumor Assays/methods
19.
Toxicol Sci ; 159(2): 436-448, 2017 10 01.
Article in English | MEDLINE | ID: mdl-28962521

ABSTRACT

Skin tumorigenesis results from DNA damage, increased inflammation, and evasion of apoptosis. The peroxisome proliferator-activated receptors (PPARs) can modulate these mechanisms in non-melanoma skin cancer. However, limited data exists regarding the role of PPARs in melanoma. This study examined the effect of proliferator-activated receptor-ß/δ (PPARß/δ) and PPARγ on cell proliferation, anchorage-dependent clonogenicity, and ectopic xenografts in the UACC903 human melanoma cell line. Stable overexpression of either PPARß/δ or PPARγ enhanced ligand-induced expression of a PPARß/δ/PPARγ target gene in UACC903 cell lines as compared with controls. The induction of target gene expression by ligand activation of PPARγ was not altered by overexpression of PPARß/δ, or vice versa. Stable overexpression of either PPARß/δ or PPARγ reduced the percentage of cells in the G1 and S phase of the cell cycle, and increased the percentage of cells in the G2/M phase of the cell cycle in UACC903 cell lines as compared with controls. Ligand activation of PPARß/δ did not further alter the distribution of cells within each phase of the cell cycle. By contrast, ligand activation of PPARγ enhanced these changes in stable UACC903 cells overexpressing PPARγ compared with controls. Stable overexpression of either PPARß/δ or PPARγ and/or ligand activation of either PPARß/δ or PPARγ inhibited cell proliferation, and anchorage-dependent clonogenicity of UACC903 cell lines as compared with controls. Further, overexpression of either PPARß/δ or PPARγ and/or ligand activation of either PPARß/δ or PPARγ inhibited ectopic xenograft tumorigenicity derived from UACC903 melanoma cells as compared with controls, and this was likely due in part to induction of apoptosis. Results from these studies demonstrate the antitumorigenic effects of both PPARß/δ and PPARγ and suggest that targeting these receptors may be useful for primary or secondary melanoma chemoprevention.


Subject(s)
Apoptosis/physiology , Inflammation/physiopathology , Melanoma/pathology , Peroxisome Proliferator-Activated Receptors/physiology , Skin Neoplasms/pathology , Animals , Cell Adhesion/physiology , Cell Cycle/physiology , Cell Line, Tumor , Cell Proliferation/physiology , Heterografts , Humans , Ligands , Mice , Mice, Nude , Peroxisome Proliferator-Activated Receptors/genetics
20.
Dev Neurobiol ; 77(9): 1057-1071, 2017 09.
Article in English | MEDLINE | ID: mdl-28245532

ABSTRACT

The mushroom bodies (MBs) are insect brain regions important for sensory integration, learning, and memory. In adult worker honey bees (Apis mellifera), the volume of neuropil associated with the MBs is larger in experienced foragers compared with hive bees and less experienced foragers. In addition, the characteristic synaptic structures of the calycal neuropils, the microglomeruli, are larger but present at lower density in 35-day-old foragers relative to 1-day-old workers. Age- and experience-based changes in plasticity of the MBs are assumed to support performance of challenging tasks, but the behavioral consequences of brain plasticity in insects are rarely examined. In this study, foragers were recruited from a field hive to a patch comprising two colors of otherwise identical artificial flowers. Flowers of one color contained a sucrose reward mimicking nectar; flowers of the second were empty. Task difficulty was adjusted by changing flower colors according to the principle of honey bee color vision space. Microglomerular volume and density in the lip (olfactory inputs) and collar (visual inputs) compartments of the MB calyces were analyzed using anti-synapsin I immunolabeling and laser scanning confocal microscopy. Foragers displayed significant variation in microglomerular volume and density, but no correlation was found between these synaptic attributes and foraging performance. © 2017 Wiley Periodicals, Inc. Develop Neurobiol 77: 1057-1071, 2017.


Subject(s)
Aging/physiology , Feeding Behavior/physiology , Mushroom Bodies/cytology , Neuronal Plasticity/physiology , Neuropil/physiology , Animals , Bees/anatomy & histology , Color Perception/physiology , Distance Perception/physiology , Learning/physiology , Male , Microscopy, Confocal , Neuropil/metabolism , Reward , Statistics, Nonparametric , Synapsins/metabolism
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