bluuebunny/crossref_metadata_2025_split huggingface dataset:
Histone Deacetylase 3 Supports Cystathionine β‐synthase Mediated Bone formation by Controlling Cytokine Signaling and Matrix Remodeling
Authors: Behera Jyotirmaya , Zhai Yuankun , Bala Jyoti , Tyagi Suresh C , Tyagi Neetu 
Published: November 2023

Cystathionine beta‐synthase (CBS) is a crucial enzyme in homocysteine (Hcy) metabolism. CBS deficiency cause hyperhomocysteinemia (HHcy) which confers diverse clinical manifestations, notably characteristic skeletal abnormalities. However, several clinical trials demonstrated that despite reduction in Hcy levels, bone disease outcome remained unaffected, thus the mechanism of bone formation is poorly defined. Therefore, we hypothesize that CBS as a novel molecular regulator in osteoblasts which promotes bone development. To test this hypothesis, we used 8–12 weeks old mice in our study; wild type mice (C57BL/6, WT) and CBS+/− mice. Silencing CBS function in osteoblast by either inhibitor hydroxylamine (HA) or CRISPR/CAS‐9 mediated gene knockout in vitro and bone marrow derived CBS‐deficient (CBS+/−) osteoblast causes reduced osteoblast differentiation and mineralization and increased osteoclast activity. Bone marrow derived CBS‐deficient (CBS+/−) osteoblasts exhibited exaggerated oxidative stress mechanism and further increased the expression of inflammatory cytokine and matrix‐degrading genes (IL‐6, MMP‐9, MMP‐13) and reduced expression of genes related to extracellular matrix production, bone development (ALP, Col1A, RUNX2, OSTERIX, OCN, OPN). ELISA assay identified among others IL‐6 to be present in plasma of CBS(+/−) mice but not in plasma of wild type mice (C57BL/6, WT). Furthermore, histone deacetylase activity 3 also decreased under oxidative stress mechanism via phosphorylation dependent manner. This leads to activation and acetylation of nuclear factor Kb (NF‐kB). This increased inflammatory cytokine production further suppresses the osteoblast differentiation and paracrine activation of osteoclasts and bone resorption via IL‐6 signaling. This increased resorptive activity is further confirmed with immunostaining of tibia cross section by TRAP‐activity and increased osteoclasts number in bone tissue of CBS+/− mice. Blockade of HDAC3 activation in CBS‐deficient (CBS+/−) osteoblast by RGFP566 (HDAC3 Inhibitor) leads to the remodelling of histone landscapes in genome and thereby accelerates histone acetylation via recognising the acetylated lysine residue in histone (H3K27ac) of chromatin. This eventually resulted in transcriptional activation of MMP9,‐13 and IL‐6 gene expression. Thus, CBS controls the temporal and spatial expression of tissue‐remodeling genes and inflammatory responses in osteoblast to ensure proper bone development. In conclusion, we demonstrate that CBS control the osteoblasts balances, osteoblast differentiation/mineralization and osteoclastgenesis via HDAC3 dependent manner during bone development.Support or Funding InformationThis work is financial supported from National Institute of health grant AR‐067667 and HL‐107640‐NT are greatly acknowledged.
Bone Marrow Transfer from Rats Fed Low or High Salt Diets Does Not Influence Salt‐Sensitive Blood Pressure and Kidney Disease in Dahl SS Recipient Rats
Authors: Fehrenbach Daniel J , Petrova Galina , Lund Hayley , Abais‐Battad Justine M , Dasinger John Henry , Mattson David L 
Published: November 2023

Increasing evidence indicates a role for the immune system in the development of hypertension and renal disease. Studies in our laboratory have demonstrated that pharmacological or genetic inhibition of adaptive immunity attenuates salt‐sensitive hypertension in the Dahl SS rat. Specifically, Dahl SS rats with a mutation in CD247 (SSCD247−/−), a gene encoding the CD3 zeta chain, lack CD3+ T‐cells and have a marked attenuation of salt‐sensitive hypertension and renal damage. The present study was designed to investigate the potential role of memory cells in the bone marrow in the development of salt‐sensitive disease. Age‐matched Dahl SS rats were kept on either a low salt (0.4% NaCl) or a high salt (4.0% NaCl) diet for 3 weeks starting at 3 weeks of age. Bone marrow from these SS rats was then transferred into age‐matched, SSCD247−/− rats (n=4 low salt bone marrow; n=5 high salt bone marrow) that underwent total body irradiation (TBI) at a dose of 11 Gy and a rate of 1.83 Gy/min. The recipient SSCD247−/− rats that underwent TBI had been maintained throughout life on the low salt diet and received bone marrow from SS rats fed the low (LSBM) or high salt (HSBM) diet. Any T cells reconstituted in the SSCD247−/− rats which underwent TBI therefore arose from the donor SS. After two weeks of recovery, the LSBM and HSBM rats were instrumented for blood pressure measurement by telemetry. While maintained on a low salt diet, there was no difference in the MAP or urine albumin excretion (Ualb) rate in the LSBM (108±2 mmHg and 0.8±0.3 mg/day) or HSBM rats (111±4mmHg and 2.4±1.6 mg/day). The rats were then switched to a high salt diet and followed for 7 days. When the rats were switched to a high salt diet, MAP and Ualb tended to increase compared to baseline in the LSBM group (127±5 mmHg and 41.4±13.5 mg/day) and MAP but not Ualb significantly increased in the HSBM group (129±6 mmHg and 60.9±27.7 mg/day), but there were no significant differences between groups after 7 days. A subsequent flow cytometry analysis demonstrated reconstitution of T‐lymphocyte (CD45+/CD3+), helper T cell (CD4+), cytotoxic T cell (CD8+), and T‐memory cell (CD44h) populations, but there were no significant differences between the two groups at this time in the any of the tissues collected (PBMCs, spleen, bone marrow, and kidneys). Though significant differences were not detected between groups in the time frame of this experiment, we plan to repeat this experiment for an extended high salt period as previous work has demonstrated the effects of immune cells on hypertension and renal damage in Dahl SS rats typically begins at approximately Day 14 of high salt.Support or Funding InformationSupported by DK96859, HL116264, and 15SFRN2391002
Effects of Whole Milk and Full‐Fat Dairy Products on Blood Pressure in Patients with Elevated Blood Pressure
Authors: Roy Stephen , Baker Brett , LaPierre Stephanie , Geary Wesley , Delfausse Laura , Pasha Evan , Tanaka Hirofumi 
Published: November 2023

We have previously demonstrated that the solitary addition of nonfat dairy products to the normal routine diet was capable of lowering blood pressure. Currently, it is not known if whole milk and full‐fat dairy products could exert similar effects as most of the available cohort studies reported no association between full‐fat dairy products and blood pressure. Therefore, the purpose of the study was to determine the efficacy of conventional full‐fat dairy products as part of the normal routine diet for reducing blood pressure in otherwise healthy adults with elevated blood pressure. After a 2‐week run‐in period, 35 subjects (13 men and 22 women) with pre‐ or stage I hypertension underwent a randomized controlled cross‐over dietary intervention trial encompassing 4 servings of dairy (whole milk, cheese, or yogurt) per day and 4 weeks of an isocaloric non‐dairy control (coconut milk, orange juice, peanuts, and applesauce) per day. All the dairy products as well as isocaloric non‐dairy products were provided in daily packages by the investigators. Two weeks of washout period was inserted between the dietary conditions. Body weight did not change throughout the experimental period. Casual (office) systolic blood pressure decreased slightly but significantly in both the dairy (135±1 vs. 131±2 mmHg) and the control (136±2 vs.133±2 mmHg) conditions. There were no significant changes in casual diastolic or mean blood pressure. Ambulatory (24 hour) systolic blood pressure did not change significantly in both dairy (131±2 vs. 130±2 mmHg) and control (131±3 vs. 129±2 mmHg) conditions. Similarly, daytime and nighttime blood pressure did not change significantly in both conditions. These data suggest that unlike non‐fat milk and non‐fat dairy products, whole milk and full‐fat dairy products added to the normal routine diet did not exert hypotensive effects in patients with elevated blood pressure.Support or Funding InformationSupported by the Dairy Research Institute
High‐Intensity Interval Training With or Without Post‐Exercise Milk Consumption Improves Cardiovascular Function in Patients with Type 2 Diabetes
Authors: Francois Monique Emily , Durrer Cody , Little Jonathan 
Published: November 2023

High‐intensity interval training (HIIT) can improve several aspects of cardiometabolic health that are important for older adults with type 2 diabetes (T2D), including cardiorespiratory fitness, body composition and endothelial function. Previous studies have suggested that adaptations to exercise training can be augmented with post‐exercise milk or protein consumption but whether this nutritional strategy can impact cardiovascular adaptations to HIIT in T2D is unknown.PurposeTo examine if cardiovascular adaptations to HIIT can be enhanced by post‐exercise milk supplementation in patients with T2D.MethodsA proof‐of‐concept double‐blind controlled trial randomized 50 adults with physician‐diagnosed T2D (58 ± 9 y, A1c: 7.1 ± 0.8%, BMI: 35 ± 7 kg/m2) to one of three nutritional beverages (500 mL skim milk, milk‐protein isolate or flavored water placebo, n=16–17 per group) consumed after exercise training (2× cardio‐ and 1× resistance‐based HIIT per week for 12 weeks). HIIT progressed from four (week 1) to 10 (weeks 6–12) 1‐min intervals of vigorous effort (eliciting ~85–90% maximal heart rate [HRmax]) interspersed with 1‐min rest/recovery. Resting HR, blood pressure (MAP), brachial artery flow‐mediated dilation (%FMD) and carotid‐femoral pulse‐wave velocity (PWV) were measured before and 72 h after the intervention. Cardiorespiratory fitness (V̇O2 peak) and HRmax were measured via incremental cycling test to exhaustion with expired gas analyses before and after training. Data were analyzed using general linear mixed models (SPSS) with magnitude‐based inferences for clinically meaningful changes assessed using the spreadsheet available at http://www.newstats.org/.ResultsThere were main effects of time (p<0.05) but no group × time interactions (all p>0.31). All results are therefore presented as main effects of time with nutritional beverage groups collapsed. HIIT lowered resting HR by −2.6 bpm (95% CI: −4.5 to −0.8 bpm), MAP by −5.7 mmHg (CI: −7.8 to −3.6 mmHg) and PWV by −1.3 m/s (0 to 2.6 m/s); the reduction in PWV was possibly and reduction in MAP was most likely clinically beneficial. HIIT increased %FMD by 1.5% (CI: 0.9 to 2.0%) and V̇O2 peak by 2.5 mL/kg/min (CI: 1.8 to 3.2 mL/kg/min); the increase in %FMD was very likely and the increase in V̇O2 peak most likely clinically beneficial. Body mass (CI: −1.8 to 0.2 kg) and HRmax (CI: −2.0 to 3.3 bpm) were not significantly changed.ConclusionTwelve weeks of HIIT significantly improves important indices of cardiovascular health in patients with T2D. Clinically meaningful improvements in blood pressure, endothelial function and cardiorespiratory fitness were evident after just 12 weeks of progressive interval training (totaling 48 – 78 minutes of exercise per week). The addition of post‐exercise milk or milk protein supplementation did not influence any of the cardiometabolic health markers assessed.Support or Funding InformationDairy Farmers of Canada.
Mesenchymal Stem Cells Derived Exosomes Promotes Bone Regeneration through Cystathionine‐β‐synthase Enhanced Angiogenesis and Osteogenesis during Bone Development
Authors: Behera Jyotirmaya , Zhai Yuankun , Bala Jyoti , Tyagi Suresh C , Tyagi Neetu 
Published: November 2023

It is critical that bone formation and angiogenesis are tightly coordinated during bone development and fracture healing, but the molecular regulator of such intercellular communication in bone microenvironment are not well studied. Cystathionine‐β‐synthase (CBS), an enzyme in homocysteine (Hcy) metabolic pathway play crucial role in osteoblast differentiation. Therefore, we hypothesize that CBS as a novel molecular regulator in osteoblasts, which regulates vascularization during bone development in osteoblast derived exosome. To test this hypothesis, we have used 7–10 weeks old male mice in our study; wild type mice (C57BL/6, WT) and CBS+/− mice. Blocking the CBS function was done by either inhibitor hydroxylamine (HA) or SiRNA prevented osteoblast differentiation and mineralization. This was supported by studies using osteoblasts cultured from bone marrow of CBS deficient (CBS+/−) mice. Exosomes of mesenchymal stem cell derived osteoblasts cultured stimulated endothelial migration and angiogenesis, which was prevented by blocking CBS in osteoblasts using HA or SiRNA. Mass spectrometry analysis and ELISA assay identified among others vascular endothelial growth factor (VEGF)‐A and VEGF homolog placental growth factor (PIGF) to be present in control osteoblasts culture medium and CBS(+/+) mice osteoblasts but not in osteoblasts treated with HA and CBS+/− osteoblasts. Immunostaining and metatarsal angiogenesis assay showed retarded vascularization in bone tissue of CBS+/− mice. However, osteopenia phenotype was confirmed in CBS+/− mice by microCT analysis. CBS+/− mice showed significantly reduced bone mineral density (BMD) and bone volume/tissue volume (BV/TV) and showed increased plasma Hcy levels compared to CBC+/+ mice (WT). However, CBS+/− mice exhibited a more significant increase in the number of TRAP‐positive osteoclasts and higher levels of RANKL than CBS+/+ mice. This suggests that Hcy levels may be associated with osteoclast activity. Furthermore, real‐time PCR and western blot analyses revealed significant decreases Alkaline phosphatase (ALP) and Runt‐related transcription factor 2 (RUNX2) expression in CBS‐deficient (CBS+/−) mouse bone marrow cells. In conclusion, we demonstrate that CBS in osteoblasts is at the crossroad of osteoblast differentiation/mineralization and angiogenesis. These findings uncover the previously undefined molecular understanding of CBS that promote angiogenesis and osteogenesis in bone development and fracture repair.Support or Funding InformationThis work is financial supported from National Institute of health grant AR‐067667 and HL‐107640‐NT are greatly acknowledged.
Serum Adipokines and Metabolic Syndrome Risk Factors in Hispanic Children
Authors: Peterson Jonathan M , Clark William Andrew , Marrs Jo‐Ann , Alamian Arsham 
Published: November 2023

Childhood obesity is a relatively new but growing epidemic in the Unites States, with 1 in 5 children currently classified as obese. Obese children are at a significantly higher risk for developing many types of cancers, cardiovascular disease, diabetes and Metabolic Syndrome (MetS). This project is focused on understanding the associations between adipose tissue health and adipose tissue‐derived secreted factors/hormones/cytokines (hereafter referred to as adipokines) and obesity and MetS in young children. Adiponectin, Leptin, and C1q TNF Related Protein 3 (CTRP3) are all adipokines which have been linked to human health, but have not been explored in children with MetS.METHODSAnthropomorphic measurements and blood collections were performed at a well child visit for Hispanic children aged 2–10 years. Lipid panel was completed at a clinical laboratory facility. Multiplex analysis (Bio‐Plex Multiplex System) was used to measure adiponectin, insulin, and leptin level. Circulating CTRP3 levels were determined by ELISA (enzyme‐linked immunosorbent assay). Risk factors for MetS were established as follows: abdominal obesity, elevated blood pressure, elevated triglycerides, low HDL and elevated plasma glucose concentrations. Pearson correlation coefficients were calculated for continuous variables and Spearman rank‐order correlation was performed for categorical variables to evaluate associations between adipokines and risk factors for MetS.RESULTSBriefly, 44 of 114 Hispanic children had 2 or more risk factors for the development of MetS. The association between Adiponectin levels, glucose, insulin, total triglyceride and Body Mass Index (BMI) trended towards significance (P>0.08), whereas leptin was significantly positively associated with BMI. CTRP3 levels were not associated with any individual measure variable, but were negatively associated with the presence of 2 or more risks factors for MetS.CONCLUSIONAdipose tissue secretes many bioactive molecules that circulate in blood, and dysregulation of adipose tissue endocrine function may promote the occurrence of MetS. The data presented in this project demonstrate the association between adipokines and MetS.Support or Funding InformationThis research was supported in part National Institute On Alcohol Abuse And Alcoholism of the National Institutes of Health under Award Number R03AA023612, Tennessee Board of Regents Diversity Research Grant (2014–2015), and by the East Tennessee State University Research Development Committee (E82262).
TNFα exposure induces neuroinflammation and insulin resistance in a rat‐derived hypothalamic cell model, rHypoE‐7
Authors: Clemenzi Matthew N , Aljghami Makram E , Wellhauser Leigh , Belsham Denise D 
Published: November 2023

Obesity is approaching epidemic levels. One of the main characteristics of obesity is the development of insulin resistance, leading to other comorbidities, such as type 2 diabetes mellitus (T2DM) and cardiovascular disease. Insulin resistance has been shown to be induced with obesity in peripheral tissues, such as liver, muscle, and adipocytes, by the pro‐inflammatory cytokine tumor necrosis factor alpha (TNFα). In the hypothalamus, the main brain region for energy regulation, neuroinflammation‐induced insulin resistance has been demonstrated to occur upon prolonged treatment with high levels of insulin and palmitate, and we are exploring whether TNFα, a downstream surrogate for palmitate, has similar effects. We hypothesized that TNFα‐treated rat hypothalamic neurons would exhibit cellular neuroinflammation and insulin resistance as evidenced by increased expression of pro‐inflammatory genes and attenuated phosphorylation of Akt (pAkt) in the insulin signaling pathway. Using an immortalized rat hypothalamic cell line (rHypoE‐7), a well‐characterized NPY‐expressing neuronal line derived from male embryonic hypothalamii, we studied changes in pAkt and inflammatory gene marker mRNA expression after TNFα treatment by Western blot and quantitative real‐time PCR, respectively. A qPCR array of 84 inflammatory markers (receptors, chemokines, and cytokines) demonstrated an increase in the expression of a number of genes encoding inflammatory markers, including Tollip and Caspase‐1, upon exposure to 100 ng/mL TNFα for 4 h. Neurons pre‐exposed to TNFα (50 ng/mL) for 6 or 16 h exhibited a significant reduction in pAkt compared to control after insulin treatment. TNFα also significantly increased mRNA expression of TNF, IK‐κB, Tollip, Tnfrsf1a, and IL6 at 4 h. In conclusion, pre‐treatment with the inflammatory cytokine TNFα causes cellular insulin resistance in the rHypoE‐7 neurons, consistent with effects seen with TNFα in peripheral tissues. It also mimics insulin‐ and palmitate‐induced insulin resistance in hypothalamic neurons. Further, these results demonstrate that components of the IKK‐β/NF‐κB pathway appear to be activated upon exposure to TNFα, providing a putative target to reverse neuroinflammation and insulin resistance with anti‐inflammatory agents.Support or Funding InformationThis research was supported by grants from the CIHR, CRC, CFI, NSERC, APS and BBDC.
PREFERENTIAL RENAL SYMPATHETIC HYPERACTIVITY IN EXPERIMENTAL DIABETES MELLITUS
Authors: Oliveira Tales L , Shimoura Caroline G , Lincevicius Gisele S , Garcia Michelle L , Boim Mirian A , Gomes Guiomar N , Bergamaschi Cassia T , Campos Ruy R 
Published: November 2023

Autonomic, cardiovascular and renal dysfunction has been observed in diabetic patients and animal models of diabetes mellitus (DM), including the streptozotocin‐induced type 1 diabetes model. The aim of this study was to evaluate the effects of the experimental DM on efferent post‐ganglionic sympathetic nerve activity, arterial baroreceptor reflex sensitivity to heart rate (HR), renal (rSNA) and lumbar (lSNA) sympathetic nerves in DM rats. The DM was induced by Streptozotocin (60 mg/kg, ip) 4 weeks before experimental analysis. Male Wistar rats (n=6–8 per group), 8 weeks of age, 230–250 g of body weight were used. The animals were separated into control (CTR) and diabetic (DM) groups. Protocol CEUA/UNIFESP 1787250714. The results were expressed as mean ± SEM and compared with t‐Student test (p<0.05). DM was confirmed by the reduction in body weight (CTR: 320.1±6.3; DM: 242.1±6.7 g, p<0.05) and significantly increased in blood glucose (CTR: 100.3±3.7; DM: 509.1±14.4 mg/dL, p<0.05) in the DM group. A significant increase in feed intake (CTR: 75.8±3; DM: 158.3±6.4 g/kg), water intake (CTR: 98±4.3; DM: 608±31,3 ml/kg), and urinary flow in the DM rats evaluated for 24h (CTR: 40±3.1; DM: 562±25,1 ml/kg) was found. Sodium urinary excretion (CTR: 5.1±0.5; DM: 17.4±1.2 mmol/kg), potassium (CTR: 16±0.9; DM: 36.4±1 mEq/kg), glucose (CTR: 12±1.4; DM: 2719±192 mg/kg), and albumin urinary excretion (CTR: 82.3±8; DM: 394.3±42.7 mg/kg) were significantly increased in DM group. In addition, sodium (CTR: 123±1.7; DM: 141.4±1.6 mmol/L) and potassium plasma concentration (CTR: 4.4±0.1; DM: 5.2±0.1 mEq/L) were significantly increased in DM rats compared to control group. The creatinine clearance was reduced in the DM rats (CTR: 1.4±0.1; DM: 1±0.1 mL/min). DM reduced heart rate (HR) (CTR: 352.6±013.5; DM: 292.2±11 bpm, p<0.05), but did not change basal values of blood pressure (BP) compared to control group. rSNA was significantly increased (CTR: 85.5±3.1; DM: 133.5±2.5 pps) but lSNA did not change in DM rats. The arterial baroceptor function was significantly decreased to rSNA, (CTR: 1.3±0.1; DM: 0.9±0.1 pps/mmHg), lSNA (CTR: 1.4±0.1, DM: 0.8±0.1 pps/mmHg) and HR control (tachycardic response = CTR: −1.4 ± 0.1; DM: 0.8 ± 0.05 and bradycardic response = CTR: 1.3 ± 0.05; DM: 0.7 ± 0.03 bpm/mmHg, respectively) in the DM group compared to control rats. Taken altogether, the results suggest that autonomic dysfunction, characterized by impaired baroreceptor function, preferential increasein renal sympathetic nerve activity associated with impaired renal function are involved in the development and/or maintenance of experimental DM. Whether the renal sympathoexcitation in DM is a cause or a consequence of renal dysfunction remains to be determined.Support or Funding InformationCAPES and FAPESP.