bluuebunny/crossref_metadata_2025_split huggingface dataset:
Cytochrome c Release from Dynamic Mitochondria in Endothelial Cells under Flow
Authors: Hu Ying‐Li , Hur Sung Sik , Chien Shu 
Published: November 2023

Endothelial mitochondria are essential to the functional integrity of endothelial cells (ECs) as they integrate a wide range of cellular processes. Cytochrome c (Cyt c) is located in the mitochondrial inner membrane space (IMS), where it functions in the electron transport chain. The morphology of mitochondria changes dynamically due to their frequent fusion and fission in response to cellular conditions. Mitochondrial fission is an early event during apoptosis and it occurs very close in time to Cyt c release, and fragmented mitochondria can readily be seen retaining Cyt c. ECs were subjected to oscillatory and laminar flows, with shear stresses of 0.4±4 and 12 dynes/cm2, respectively. Cyt c staining of the mitochondria was detected in ECs under both types of flow. In comparison to static control, Cyt c was significantly reduced in the mitochondria of ECs exposed to both flow patterns for 4 h, 8 h, and 24 h. The reduction of Cyt c in endothelial mitochondria was greater under oscillatory flow for 8 h and 24 h.

4 h
8 h
24 h

Oscillatory Flow/Static
0.16±0.05
0.25±0.06*
0.59±0.05*

Laminar Flow/Static
0.19±0.03
0.44±0.03
0.90±0.03

Values are mean±SEM.
P<0.05 for Oscillatory vs. Laminar Flow.

After laminar flow for 24 h, Cyt c rose to almost the level of the static ECs. We used time‐lapse imaging to monitor the spatial‐temporal dynamics of mitochondria in live ECs transfected with RFP‐mito. During live ECs shearing process mitochondria networks are dramatically reorganized from long filamentous, interconnected tubules into small punctate spheres in ECs under both flow patterns. Under oscillatory flow, more small fragmentations of endothelial mitochondria were observed than those under laminar flow. The results suggest that mechanical force affects the mitochondria dynamics in ECs under flow to cause Cyt c release, which was greater under oscillatory than laminar flow. Understanding of the endothelial mitochondria responses to different flow patterns will help to elucidate the mechanism of the region‐specific localization of atherosclerosis in the arterial system.Support or Funding InformationThis work was supported by NHLBI Research Grants HL‐104402 and HL‐106579 (S.C.)
Gene Polymorphism of Matrix Metalloproteinases 3, and 9 in Breast Cancer
Authors: Soliman Naglaa Raafat Abdraboh , Bayoumi Fatehia Ali 
Published: November 2023

BackgroundGenetic factors have a considerable influence on development and progress of malignancy. Matrix metallo‐proteinases (MMPs) are a group of proteolytic enzymes which are proved to be involved in pathogenesis of breast cancer. Some functional single nucleotide polymorphisms (SNPs) affect the transcriptional activity of genes of MMPs. We aimed in the present study to investigate the possible association of 2 SNPs, C/T polymorphism of MMP9 gene, and 5A/6A polymorphism of MMP3 gene, with the occurrence and clinico‐pathological criteria of breast cancer.Subject and methodThe present case control study included 59 breast cancer female patients, and 77 tumor free age matched control females. The SNPs were genotyped using polymerase chain reaction ‐ restriction fragment length polymorphism assay.ResultsData analysis have shown higher frequency of T allele of C/T MMP9 and A5 of A5/A6 MMP3 in breast cancer patients as compared to control subjects with odd ratio (OR), 95%confidence interval (95% CI) 1.8 (1.02 – 3.26) and 1.2 (0.71 – 1.89) respectively. When the frequency of both alleles was analyzed in comparison to prognostic clinico‐pathological criteria of the disease, there was a significant association of T allele of C/T MMP9 with advanced tumor size and histological grade with OR (95%CI) 2.25 (1.12 – 5.64), 1.8 (0.78 – 4.17) respectively.Conclusionthe results of the current study suggested that T allele of C/T MMP9 gene polymorphism is strongly linked to the incidence, growth and differentiation of breast cancer.Support or Funding InformationFund supplied by Dubai Medical College
Estrogen Receptor α induces microRNA‐365 expression to attenuate classical macrophage activation
Authors: Chang Richard Cheng‐An , Tseng Alexander Mike , Wang Haiqing , Lin Yu‐Lieh , Zhou Beiyan 
Published: November 2023

The sex steroid hormone estrogen, besides its role in reproduction, has been reported as a potential regulator for immune responses. Albeit the increasing co‐morbidity associated with obesity, the pandemic threat marked with chronic inflammation and insulin resistance, only a handful of studies suggest estrogen may exert immune suppressive effect in this context. In this study, we report the first set of evidence to support that estrogen receptor alpha (ERα) signaling can selectively suppress classical activation of macrophages, which is the primary contributor to obesity‐induced chronic inflammation.First, our microRNA array analysis from macrophages subjected to polarized activation conditions revealed a set of microRNAs were actively regulated in macrophage classical activation. Bone marrow‐derived macrophages (BMDMs) stimulated with lipopolysaccharide/interferon gamma (LPS/INFγ) inflammatory responses (M1) and alternative activation (M2) by IL‐4/IL‐13 treatment. There were 33 miRNAs detected in naïve macrophage (M0), 156 miRNAs detected in M1, and 159 miRNAs detected in M2. Furthermore, there were 33 miRNAs displayed significant differential expression patterns in M1 or M2. Genomic analysis by surveying the promoters of these miRNAs identified a cluster of 10 miRNAs bearing ERα binding sites within 10 upstream kilobases. Interestingly, BMDMs subjected to 17β‐estradiol treatment exhibited suppressed M1 responses accompanied by significantly higher levels of ERa and miR‐365; whereas no detectable changes in ERa level in M2 activation. BMDMs treated with LPS/INFγ in the presence of 17β‐estradiol exhibited significantly lower levels of IL‐6, TNFα, and IL‐1β. Of note, miR‐365 level was significantly lower in M1 compared to M0, which was reversed by 17β‐estradiol treatment. Validation of the regulatory axis of ERa‐miR365 in macrophage classic activation will provide crucial evidence to support a novel paradigm shift in understanding the function of estrogen in obesity‐induced tissue inflammation and subsequent systemic disorders.Support or Funding InformationAmerican Diabetes Association (1‐13‐JF‐59 to B. Zhou) and National Institute of Health/National Institute of Diabetes and Digestive and Kidney Diseases (NIH/NIDDK 1R01DK098662 to B. Zhou)
Physiological And Transcriptomic Analyses In Escherischia coli Reveal Effects On Stress Associated Metabolism In The Presence of Oxidized Graphene
Authors: RAMPAL SAKSHI , Barabote Ravi Damodar 
Published: November 2023

Graphene is a carbon nanomaterial that has a great potential to be used in wide variety of fields especially in biomedical applications. However, the environmental and health effects of graphene‐derived nanomaterials are not fully understood. Oxidized graphene is widely used in the synthesis of graphene‐derived nanomaterials. We assessed the biological effects of oxidized graphene using Escherichia coli as a model bacterium. Growth of E. coli was monitored in the presence of pristine graphene and oxidized graphene using dilution plating and spot test analysis. Growth analysis revealed that addition of oxidized graphene at concentrations greater than 80 μg/ml inhibited the growth of E. coli. To understand the molecular response of E. coli to graphene, RNA‐seq technology (Illumina platform) was used. Transcriptomic analysis of E. coli exposed to pristine graphene and oxidized graphene revealed several genes that were substantially differentially expressed. The genes that showed higher expression in the presence of oxidized graphene encode for stress response proteins, membrane proteins, transporters, acid resistance proteins, and proteins involved in sulfur metabolism. These data suggests that oxidized graphene likely elicits oxidative stress in E. coli. Using gene deletion knockouts, we studied the physiological role of a few of the genes involved in above‐mentioned functions. Deletion mutants of an inner membrane protein and a sulfite reductase were grown in the absence or presence of oxidized graphene. Growth data showed that both genes are critical for the bacteria to survive under stress conditions induced by oxidized graphene. In addition, we are studying the expression responses of select genes using GFP reporter constructs to identify biomarker genes specific to oxidized graphene.Support or Funding InformationDepartment of Biological Sciences, University of Arkansas; Cell and Molecular Biology Program, University of Arkansas; Arkansas Research Alliance
Up‐regulating Heme Oxygenase System Potentiates Pancreatic Repair by Enhancing Proteins of Regeneration
Authors: Mishra Manish , Ndisang Joseph Fomusi 
Published: November 2023

BackgroundAutoimmune‐mediated enhanced beta cell destruction and resultant insulin insufficiency poses a great health risk in type‐1 diabetes mellitus affected individuals. Our previous observations have shown that up‐regulation of the heme oxygenase (HO) by HO‐inducer hemin improved pancreatic lesion in streptozotocin‐induced diabetes, yet the hemin mediated pancreatic tissue repair mechanisms are still unclear. Accordingly, the aim of this study was to examine the effects of up‐regulating the HO system on proteins of regeneration during pancreatic repair in streptozotocin‐induced type‐1 diabetic animal model.Methods/ResultsStreptozotocin (STZ) was used to induce diabetes, while HO system was up‐regulated with hemin or inhibited with chromium mesoporphyrin. Histology/morphology, spectrophotometry, enzyme‐immunoassay (EIA), enzyme‐linked immunosorbent assay (ELISA) and Western‐immunoblotting were used to demonstrate pancreatic tissue injury and repair.Our data demonstrated that treatment with hemin to STZ‐induced diabetic animals effectively suppressed the pancreatic histopathological lesions (vacuolization, interstitial edema, mononuclear cell infiltration and fibrosis), while enhanced several proteins implicated in pancreatic repair/regeneration and beta cell growth and maturation including Oct3/4, Pax‐2, beta‐catenin, ISL‐1, glucose transporter‐2 (GLUT2), homeodomain protein NKK6.1. Interestingly, hemin treatment also increased the expression of stem cell markers such as cKit and Sca‐1, suggesting the involvement of stem cells and proteins of regeneration during pancreatic repair/regeneration in STZ diabetic animals. Moreover, the restoration of pancreatic morphology was accompanied by increased production of insulin and the potentiation of important proteins of insulin signaling pathway such as insulin‐receptor substrate (IRS‐1), IRS‐2, phosphatidylinositol‐3‐kinase (PI3K) and Glucose transporter‐4 (GLUT4) in skeletal muscles and the liver from STZ diabetic animals. Furthermore, hemin significantly reduced the inflammation and oxidative stress in the pancreas, skeletal muscles and liver, In contrast, these effects were nullified by the HO‐inhibitor, chromium mesoporphyrin.ConclusionTaken together, our findings demonstrate that upregulating the HO‐system with hemin is a useful strategy to potentiate proteins of regeneration such as Oct3/4, Pax‐2, beta‐catenin, ISL‐1, GLUT2 and NKK6.1 during the restoration of pancreatic morphology in streptozotocin‐induced diabetic animals. Importantly, the restoration of pancreatic morphology was accompanied by improved insulin production and the potentiation of proteins implicated in insulin signaling and glucose metabolism. Thus, HO‐inducers may open a new horizon in the search for remedies of type‐1 diabetes.Support or Funding InformationHeart & Stroke Foundation of Saskatchewan, Canada
Measurement of Nitric Oxide with EPR in Whole Blood during Peak Hand Grip Exercise in Women with Metabolic Syndrome
Authors: Barlow Matthew A , Cheeti Upender R , Navarro Juan P , Stephens Kyle , Weaver John M 
Published: November 2023

The contribution of bioavailable Nitric Oxide to the change in vascular function during exercise hyperemia is primarily concluded through the lack of functional changes in the presence of eNOS inhibitors. The aim of this study is to utilize EPR techniques to measure the changes in NO production during peak hand grip exercise to identify mechanistic differences of vascular function in women with Metabolic Syndrome compared to healthy controls.MethodsIn this study, 13 participants (5 MetSyn and 8 Controls) performed the procedure of graded handgrip exercise on a dynamic handgrip device while brachial blood flow was continuously measured with Doppler ultrasound and B‐mode (GE Vivid E with 6MHz linear probe). Following a brief rest, individuals were asked to perform handgrip exercise whereby the workload progressively increased in a ramp fashion (0.5 kg·min−1) until the required contraction rate (30 cpm) could no longer be maintained. At rest and immediately after halting exercise at task failure for individual peak resistance, whole blood was drawn into a prepared vaccutainer in a 1:1 venous blood to the metal chelator, defferoxamine (DF), in diethyldithiocarbamate (DETC) Krebs buffer. The samples were immediately flash frozen in liquid nitrogen and stored at −80 degrees Celsius. All pre and peak NO samples were measured and compared with known NO samples using an X‐band electron paramagnetic resonance spectrometer (Bruker Biospin, Karlsruhe, Germany). Whole blood with known concentrations of NO2 (1–10mM) and Na2S2O4 (20 mM) served as a reference.ResultsWhole blood NO increased significantly from rest to peak exercise in both groups (p=0.047) in addition to brachial conductance (p=0.0001). There was no significant difference in the rise in NO during peak exercise between the MetSyn and Control groups (p=0.67) however there is a significant difference in brachial conductance starting, peak and delta between the groups (p=0.021, 0.028, and 0.019).ConclusionThe whole blood Iron Chelated EPR technique is capable of detecting the degree of change in NO potentially contributing exercise hyperemia in different groups and potentially at variable grades of exercise. Although there is a significant difference in the vascular conductance between the groups, with the number of participants included in this study, there was no difference indicated between the groups for NO at rest and peak exercise.Support or Funding InformationResearch reported in this publication was supported by an Institutional Development Award (IDeA) from the National Institute of General Medical Sciences of the National Institutes of Health under grant number P20GM103451.
Precooling of fanning and hands and feet water immersion attenuates dehydration and thirst during exercise in the heat
Authors: Tokizawa Ken , Oka Tatsuo , Son Su‐young 
Published: November 2023

Precooling (i.e., removal of heat from the body immediately prior to exercise) is a popular strategy for alleviating thermal strain. Although whole body water immersion is most commonly used to precool in experimental studies, this method is not always possible in all field settings, or practical. In the present study, we examined the effectiveness of a novel combination of fanning and hands and feet water immersion as practical precooling method on heat strain while wearing protective clothing.Nine males engaged in 60 min of walking at a moderate speed (2.5 km/h) in a hot environment (37°C, 50%RH) while wearing protective clothing. Before walking, they were exposed to fanning (4 m/s) and spraying over the body and to hands and feet water immersion (18°C) for 30 min (FAN+WI).Rectal temperature and heart rate at the end of walking were lower in WI+FAN trial than in the control trial (37.7 ± 0.1 vs. 38.2 ± 0.1°C; 134 ± 9 vs. 118 ± 9°C, p < 0.05, respectively). The attenuations in FAN+WI trial did not differ to that in WI only trial (we have previously reported). Sweat rate in the chest and percentage reduction in body weight were inhibited by WI+FAN (1.3 ± 0.1 vs. 1.8 ± 0.1 mg/cm2/min; −1.6 ± 0.2 vs. −0.9 ± 0.1 %, p < 0.05, respectively), and the inhibitions were greater than in WI only trial (p < 0.05). Mean skin temperature was lower in FAN+WI trial than in the control at the end of precooling (29.0 ± 0.4 vs. 34.0 ± 0.2°C, p < 0.05) and walking (37.0 ± 0.1 vs. 37.3 ± 0.1°C, p < 0.05). Perceptions by ratings of visual analogue scale (thermal sensation and pleasantness, physical and psychological fatigues, thirst, and damp sensation) were lower in WI+FAN trial than in the control trial (p < 0.05). The attenuation in the rating score of thirst has not been seen in WI only trial.These results suggest that combined precooling of fanning and hands and feet water immersion can inhibit dehydration and thirst more compared to each cooling conducted alone. The lower mean body skin temperature in the combined cooling procedure may contribute to attenuation of sweating.Support or Funding InformationKAKENHI‐25293155
Effect of Acute Local Heating on Lower Extremity Macro‐ and Microvascular Dilator Function in Humans
Authors: Romero Steven A. , Gagnon Daniel , Cramer Matthew N. , Simmons Grant H. , Pellinger Thomas K. , Poh Paula Y.S. , Kouda Ken , Crandall Craig G. 
Published: November 2023

Local heating of an extremity increases blood flow and vascular shear stress throughout the arterial vascular tree. Local heating acutely improves macrovascular dilator function in the upper extremities through a shear stress dependent mechanism. However, the effect of local heating on macro‐and microvascular dilator function of the lower extremities is unclear. Therefore, the purpose of this study was to test the hypothesis that acute local heating improves lower extremity macro‐ and microvascular dilator function. Five healthy subjects (2 females; age 27 ± 4 years; height 166 ± 4 cm; weight 65 ± 7 kg; mean ± SD) immersed their lower extremities at a depth of ~33 cm into a circulated water bath (42.5 ± 0.4°C) for 45 min. Superficial femoral artery blood flow and diameter (Doppler ultrasound) were measured prior to and every 15 min during heating. Intestinal temperature, skin and skeletal muscle temperatures of the calf were measured. Prior to and 30 min after heating, macro‐ (flow‐mediated dilation) and microvascular (3 min reactive hyperemia area under the curve) dilator function were assessed in the superficial femoral artery following 5 min of arterial occlusion. Local heating increased superficial femoral artery blood flow (pre 63.7 ± 10.8 mL min−1 vs. 45 min local heating 606.3 ± 54.3 mL min−1) and shear rate (pre 51.5 ± 12.9 s−1 vs. 45 min local heating 410.6 ± 70.9 s−1; both P < 0.05). Intestinal temperature (Δ0.7 ± 0.1°C), calf skin temperature (Δ9.1 ± 0.1°C), and muscle temperature (Δ8.3 ± 0.6°C; all P < 0.05) increased at 45 min local heating. Macrovascular dilator function was unchanged following heating (pre 4.6 ± 1.1% vs. post 4.6 ± 0.7%; P = 0.8). Microvascular dilator function increased following local heating (pre 456 ± 89 mL vs. post 605 ± 101 mL; P = 0.02). However, after normalizing for differences in baseline blood flow, microvascular dilator function did not differ (pre 423 ± 71 mL vs. post 556 ± 91 mL; P = 0.2). Taken together, local heating of the lower extremities increases blood flow and shear rate similar to that previously observed in the upper extremities. However, these data suggest that these hemodynamic changes do not improve lower extremity macro‐ or microvascular dilator function following local heating in healthy humans.Support or Funding InformationFunded by National Institutes of Health (GM‐068865).
Age‐ and Sex‐Related Differences in the Cutaneous Vascular Response to Exogenous Angiotensin II
Authors: Krajek Alex C , Kolb Kelsey E , Lang James A 
Published: November 2023

The enzymatic machinery for the production and activation of angiotensin II (AngII) is locally expressed in human skin. Moreover, it has been demonstrated that AngII contributes to the reflex vasoconstriction (VC) response in aged skin; however, the cutaneous responsivity of AngII across a range of doses has not been examined. Thus, we hypothesized that the cutaneous VC response will incrementally increase with higher doses of Ang II. Additionally, we hypothesized that AngII‐mediated VC will be affected by the inhibition of adrenoreceptor or angiotensin II type II receptor (AT2R) pathways. Three microdialysis (MD) fibers were placed in the forearm skin of 8 young (Y: 23 ± 1 year) and 8 older (O: 68 ± 1 year) individuals for administration of 1) lactated Ringer's solution (control), 2) adrenoreceptor blockade with 5mM yohimbine + 1 mM propranolol (Y+P), and 3) AT2R inhibition with 1 μM PD‐123319 (PD). Then, AngII was added to the perfusates at eight graded dose concentrations, ranging from 0.1 nM to 1 mM, administered at 10 min intervals. Laser Doppler flux (LDF) was measured for each MD site and cutaneous vascular conductance (CVC) was calculated as CVC = LDF/mean arterial pressure and was expressed as a percent change from baseline (%ΔCVC). Compared to the control site (Y = 31 ± 6, O = 36 ± 5 %ΔCVC), the peak VC response at the highest dose was attenuated at the Y+P site (Y = 20 ± 5%, O = 32 ± 6 %ΔCVC) but was elevated at the PD site (Y = 42 ± 10, O = 41 ± 5 %ΔCVC). Furthermore, older women (44 ± 5 %ΔCVC) exhibited a greater peak VC at the control site compared to older men (23 ± 5 %ΔCVC; p < 0.05). These results indicate that older women have greater vasoconstrictor sensitivity to exogenous AngII than young individuals or older men. These data further suggest that AngII may be affecting multiple targets, including adrenergic and AT2R pathways.Support or Funding InformationDMU IOER Grant 05‐13‐05
Bicarbonate Infusion Attenuates Arterial Desaturation During Maximal Exercise
Authors: Volianitis Stefanos , Bredmose Per P , Nielsen Henning B , Strømstad Morten , Quistorff Bjørn , Secher Niels H 
Published: November 2023

Maximal exercise is associated with a reduced O2 saturation of arterial haemoglobin (SaO2). We evaluated the effect of pH in the reduction of SaO2 during maximal exercise by attenuating the metabolic acidosis with intravenous administration of bicarbonate. Eight competitive oarsmen (age 22.5 ± 3 yr, body weight 78 ± 2 kg, and height 1.83 ± 0.05 m; mean ± SD) performed 2000 m maximal ergometer row in random order either with sodium bicarbonate (Bicarb, 100 to 325 ml; 1 mM) or an equal volume of 0.9% saline (Sal). Arterial blood samples were obtained at rest and immediately after exercise. During the Sal trial arterial pH was reduced to 7.12 ± 0.01, while during Bicarb acidosis was attenuated in a dose related manner (7.29 ± 0.03; P<0.05). Lactate concentration during Bicarb was higher than Sal (26 ± 3 vs. 20 ± 1 mM; P<0.05). Also, bicarbonate concentration and SaO2 were higher (20.5 ± 1.8 mM vs 11.8 ± 0.8 mM and 92.5 ± 1% vs 89.4 ± 1%; P<0.05) during Bicarb compared to Sal, respectively. Arterial O2 partial pressure decreased similarly during the two trials (82.5 ± 2.6 vs. 87.3 ± 2.1 mmHg). The results demonstrate that during maximal exercise the decrease in SaO2 is influenced not only by the limited pulmonary diffusion capacity but also by the attenuated O2 affinity for hemoglobin induced by the decrease of plasma pH (Bohr effect).Support or Funding InformationDr. Per Bredmose received a grant from The Købmand i Odense Johann og Hanne Weimann f. Seedorfs foundation.