bluuebunny/crossref_metadata_2025 huggingface dataset:
Integrated cAMP Signaling by the Mixed Agonist WIN55212–2 via Dual Activation of CB1 and CB2 Receptors in Rat Cortical Astrocytes
Authors: Shivachar Amruthesh C , Leonard Golda Anne , Torres‐Hernandez Jose A
Published: June 2021

While cannabinoid CB1 receptor signaling is well established in astroglia, it is unclear whether cannabinoid CB2 receptors are also involved. Here we demonstrate the co‐expression and dual activation of both cannabinoid receptors by WIN 55212–2 in rat cortical astrocytes. Highly enriched astrocytes, expressing astroglial‐specific marker, glial fibrillary acidic protein (GFAP) were used for pharmacological, immunocytochemical and biochemical studies. The CB1/CB2 agonist, WIN 55212–2 (1μM), inhibited about 61% of forskolin‐induced cAMP (80.87±5.08 nmol/well). WIN 55212–2 inhibition of cAMP was attenuated either by the CB1 receptor‐selective antagonist /inverse agonist, AM 251(25 nM), or by the CB2 receptor‐selective antagonist/inverse agonist, AM 630 (100nM). However there was no additive effect when both of these antagonists were added, suggesting an integrated signaling mechanism. Immunocytochemical double‐labeling studies showed that GFAP‐positive cells also stained positive for the cannabinoid CB1 and CB2 receptors. Quantitative western blot analyses showed immunoreactive bands at 62 KDa and 40 KDa, the molecular mass of CB1 and CB2, respectively. Our findings suggest that CB1 and CB2 receptors are constitutively expressed in rat cortical astrocytes and produce an integrated response when simultaneously stimulated with a CB1/CB2 agonist (*support: Institutional seed grant).
Ganoderma lucidum Polysaccharide Accelerates Refractory Wound Healing by Inhibition of Mitochondrial Oxidative Stress in Diabetes
Authors: Tie Lu , Lin Zhi‐Bin , Li Xue‐Jun
Published: June 2021

Background

Impaired wound healing in diabetic patients is a serious complication with limited treatment regimens. Gene therapy of mitochondrial antioxidant enzyme manganese superoxide dismutase (MnSOD) was able to restore diabetic wound healing with suppression of O2.−. The present study was designed to determine protective effects of Ganoderma lucidum polysaccharide (Gl‐PS), as a potent inducer of MnSOD, on diabetic wound healing.

Method and Results

Streptozotocin‐induced diabetic mice with full‐thickness excisional wounds were administered with Gl‐PS (10, 50 or 250 mg/kg/day, i.g.). Gl‐PS dose‐dependently rescued the delay of wound closure and increased the mean perfusion rate around the wound in diabetic mice. Diabetic conditions markly increased mitochondrial O2.− production and nitrotyrosine formation in wound tissues, which were normalized with Gl‐PS treatment. In diabetic wound tissues, the protein level of MnSOD was unchanged whereas MnSOD activity was inhibited and its nitration was potentiated; Gl‐PS administration suppressed MnSOD nitration and increased MnSOD and glutathione peroxidase (GPx) activities. Moreover, Gl‐PS attenuated the redox enzyme p66Shc expression and phosphorylation in diabetic mice skin.

Conclusion

Gl‐PS rescued the delayed wound healing and improved wound angiogenesis in diabetic mice, at least in part, by suppression of mitochondrial oxidative stress. This work was supported by the National Natural Science Foundation of China.
Effect of sodium nitrite supplementation on endothelial dysfunction in diabetic mice
Authors: Reese Lauren R. , Sindler Amy L. , Cox‐York Kimberly A. , Seals Doug R. , Gentile Christopher L.
Published: June 2021

Nitrite is an oxidative breakdown product of nitric oxide that has been shown to restore cardiovascular dysfunction in various settings, although its effects on diabetes‐related endothelial dysfunction are unclear. We therefore tested the hypothesis that short‐term nitrite therapy restores endothelial dysfunction in diabetic mice. Genetically obese, diabetic mice (db/db) were administered drinking water with or without sodium nitrite (50 mg/‐L) for 4 weeks. Isolated carotid arteries of diabetic mice had lower acetylcholine (ACh)‐induced endothelium‐dependent dilation (EDD) compared to heterozygous control mice (db/+) (71.2 ±14.3% vs. 93±7.0%; p<0.05). Sodium nitrite supplementation restored EDD to control levels (92.9±2.3% vs. 93±7.0%; p<0.05). Sodium Nitroprusside (SNP)‐induced endothelium‐independent dilation did not differ between diabetic and control mice (99.8±4.2% vs. 97.2±4.2%; p<0.05) or following nitrite supplementation (99.8±1.2%; p<0.05). These data suggest that sodium nitrite may be a novel therapy for treating diabetes‐related arterial dysfunction.

5 K01 DK087777‐03 PI: Gentile, Christopher, NIH AG013038 PI: Douglas R. Seals
Aryl Hydrocarbon Receptor Inhibits Lysophosphatidic Acid‐Induced Vascular Endothelial Growth Factors Expression in Prostate Cancer Cells
Authors: Lin Yueh‐Chien , Lan Shun‐Yan , Wu Pei‐Yi , Huang Yuan‐Li , Lee Hsinyu
Published: June 2021

Lysophosphatidic acid (LPA) is a lipid growth factor with multiple biological functions. Our previous studies demonstrated that LPA induces Vascular Endothelial Growth Factors, VEGF‐A and VEGF‐C, expression in PC‐3 prostate cancer cells. Hypoxia‐inducible factor‐1 alpha (HIF‐1α) is an important regulator for angiogenesis processes through enhancing VEGF‐A expression by dimerizing with aryl hydrocarbon receptor nuclear translocator (ARNT/ HIF‐1β) to form HIF‐1 complex. Aryl hydrocarbon receptor (AhR), a receptor for dioxin‐like compounds, also dimerizes with ARNT. Since ARNT is a common dimerization partner for AhR and HIF‐1α, we hypothesized that overexpression of AhR might suppress LPA‐induced VEGFs expression in PC‐3 cells by competing with HIF‐1α for ARNT. In this study, we demonstrated the effects of AhR on LPA‐induced VEGFs expression. Over‐expression and activation of AhR inhibited LPA‐induced VEGFs expression in PC‐3 cells. In addition, LPA‐induced VEGF‐A and VEGF‐C expressions were regulated by HIF‐1α through activation of the phosphatidylinositol 3‐kinase (PI3K). These results indicate that AhR inhibits LPA‐induced VEGFs expression in PC‐3 cells by attenuating HIF‐1α signaling. This study suggests AhR might be a therapeutic target for prevention of prostate cancer metastasis.
Fluid shifts during running with body weight unloading by LBPP
Authors: Mack Gary W , Rasmussen Nicole N , Kunz Justin D , Gillespie Michael A
Published: June 2021

At any given running speed, weight support with a lower body positive pressure (LBPP) device (i.e. Alter‐G treadmill) reduces VO2. We tested the hypothesis that mild LBPP would alter plasma volume (PV) shifts during exercise. Maximal aerobic capacity (VO2max) was determined in 8 male subjects at 100% and 80% (13.5 ± 0.1 mmHg LBPP) body weight loading at 0% grade. VO2, heart rate (HR), mean arterial blood pressure (MAP) and changes in Hct, [Hb], and total protein ([TP]) were determined on separate days in a randomized crossover design. Venous blood samples (5 cc) were taken at rest and at 5, 6, 7, 7.5, 8.0, and 8.5, 9.0 mph (0% grade) during each 3 min stage. Changes in plasma volume were calculated from changes in Hct and [Hb]. VO2max was similar for both BW loading trials and averaged about 52.5 ± 0.9 ml O2•min−1•kg−1 BW. The energy cost of running at 9 mph (42.3 ± 1.7 ml O2•min−1•kg−1 BW ) was reduced by 12% at 80% body weight (p<0.05). During running at 80% BW HR was reduced compared to 100% BW running (p<0.05) however the MAP response was similar. During exercise the reduction in PV, at any given VO2 was larger at 80% BW compared to 100% BW running (p<0.001). At any give energy expenditure exercise with mild LBPP produced a greater shift of fluid out of the vascular space. It is interesting that 80% BW running resulted in a lower HR despite the larger reduction in PV. The impact of mild LBPP on venous return must counteract the role of reduced PV on stroke volume, thereby supporting a lower exercise HR.
Thermal perception in trained and untrained young men
Authors: TAKEDA RYOSUKE , Okazaki Kazunobu , Imai Daiki , Suzuki Akina , Naghavi Nooshin , Yokoyama Hisayo , Miyagawa Toshiaki
Published: June 2021

Increase in skin and esophageal temperature (Tsk and Tes) enhances autonomic and behavioral thermoregulatory responses with the perception of both temperatures. Endurance training increases autonomic thermoregulatory responses therefore we assessed if the perception of both temperatures was modified with endurance training.

Methods

Seven trained (< 3 days of training/week) and six untrained healthy young men underwent measurements of noticeable increase/decrease (± 0.1°C/sec) of skin temperature (warm/cold threshold) at chest by using a thermode (6.25 cm2), and of subjective whole body thermal sensation (visual analogue scale) in normothermia (NT, Tes; 36.6 ± 0.2°C) and hyperthermia (HT, Tes; 37.3 ± 0.1°C, lower legs immersion in 42°C water). Tes, Tsk, and cutaneous vascular conductance (CVC) and sweat rate (SR) at forearm and chest were measured continuously.

Results

In both groups, cold threshold was decreased and subjective whole body thermal sensation, CVC and SR at both sites were increased in HT compared with NT (P < 0.05), while warm threshold remained unchanged. In NT and HT, cold threshold and subjective whole body thermal sensation were similar in both groups, while warm threshold was lower (more sensitive) in trained than untrained men (P < 0.05).

Conclusions

Skin warm perception but not skin cold perception or whole body thermal sensation is more sensitive in trained than untrained men.
Effects of heat stress on mechanical ventilation‐induced atrophy in rat diaphragm
Authors: Ichinoseki‐Sekine Noriko , Yoshihara Toshinori , Kakigi Ryo , Sugiura Takao , Powers Scott K , Kawai Sachio , Naito Hisashi
Published: June 2021

This study investigated the effects of heat stress on mechanical ventilation‐induced atrophy in rat diaphragm. Adult male Wistar rats (n=38) were assigned randomly to control, heat stress, mechanical ventilation (MV), or heat stress + MV groups. Heat stress consisted of 1 h in a heat chamber set at 41°C, 24 h before MV. MV was performed for 12 h under anesthesia. Immediately after MV, diaphragm was removed, frozen and stored for analysis. All procedures were approved by the animal ethics committee of Juntendo University. MV significantly increased cleaved caspase‐3 (p<0.05) and reduced the mean muscle fiber cross‐sectional area (CSA, p<0.05). TUNEL index was not affected by MV. Heat stress resulted in a significant induction of heat shock protein72 (p<0.01), and attenuated reduction of CSA (p<0.05). Cleaved caspase‐3 and TUNEL index tended to be decreased by heat stress. These results indicate that 1 h whole‐body heat stress 24 h before MV may attenuate the acute diaphragmatic atrophy via an apoptotic pathway.

This work was supported by a Grant‐in‐Aid for Young Scientists (B) No. 23700781 and JSPS Fellows No. 11J11122 by JSPS.
Interaction between angiotensin II (AngII) and brain‐derived neurotrophic factor (BDNF) in modulating K+ currents
Authors: Becker Bryan Keith , Wang Hanjun , Zucker Irving H.
Published: June 2021

Sympathoexcitation in the setting of chronic heart failure and hypertension is a common and poor prognostic indicator during the progression of the disease. AngII has been widely implicated in the development of this pathology, however, the precise mechanisms by which AngII drives sympathoexcitation is incompletely understood. Recent evidence has emerged implicating the potential for an interaction between AngII and BDNF in the development of sympathoexcitation by increasing neuronal sensitivity. One likely mechanism leading to heightened neuronal sensitivity is the inhibition of outward K+ currents. Here, we investigated the role AngII and BDNF have on modulating K+ currents in CATH.a neuronal cell culture as measured by whole‐cell patch clamp. CATH.a neurons treated with exogenous BDNF (50ng/mL) for 2 hours exhibited decreased peak K+ currents (21.5 ± 4.8 pA/pF, p<0.01 n=6) compared to control (63.3 ± 9.7 pA/pF, n=10). Acute superfusion of 50ng/mL BDNF for 10 minutes had no significant effect on K+ currents. CATH.a treated for 6 hours with 100nM AngII exhibited decreased peak K+ currents (70.3 ± 7.4 pA/pF, p<0.001 n=10) compared to control (136.7 ± 10.1 pA/pF, n=11). Cotreatment of AngII with anti‐BDNF antibody significantly attenuated this response (106.9 ± 11.9 pA/pF, p<0.05 n=7). These results indicate that BDNF may be a potential contributor to the decreased K+ currents and increased neuronal sensitivity due to AngII and provide novel information about a potential role for BDNF in promoting sympathoexcitation.
Cross‐talk between central ACE/ACE2 and RhoA/ROCKII Pathways in Chronic Heart Failure.
Authors: Haack Karla , Xiao Liang , Zucker Irving
Published: June 2021

A hallmark of chronic heart failure (CHF) is an overactivation of the renin‐angiotensin system (RAS) and an imbalance of angiotensin converting enzyme (ACE) and angiotensin converting enzyme 2 (ACE2). This leads to increased Angiotensin II (AngII) and contributes to sympathetic hyperactivity and oxidative stress. The RhoA/ROCKII pathway is activated by AngII and is hyperactive in CHF contributing to a pro‐oxidant state. We hypothesized that there is an interaction between central RhoA/ROCKII and RAS pathways in CHF. Preliminary studies from our lab indicated in a rabbit model of CHF, central Fasudil (a ROCKII inhibitor) normalized ACE and AngII type 1 Receptor (AT1R) expression to sham levels and increased ACE2 expression in the RVLM. To further investigate the connection between these two pathways, we induced CHF in both wild type C57Bl/6 (WT) mice and in transgenic human ACE2 overexpression mouse model selectively in the brain (synhACE2). In the brainstem of CHF WT mice, there was a significant (~10 fold, p=0.03) increase in pRhoA and a 10% increase in ROCKII protein compared to sham. However in CHF synhACE2 mice, pRhoA and ROCKII were blunted compared to CHF WT (pRhoA: 3.6 ±0.5 vs 10.6 ± 1.8, p=0.06; ROCKII: 0.3±0.1 vs 1.3±0.1, p<0.05). Silencing the mas Receptor by siRNA in the CHF synhACE2 mice returned pRhoA and ROCKII to CHF WT levels. Taken together, these data suggest a cross‐talk between central RAS and RhoA/ROCKII pathways in CHF. Increasing ACE2 and Angiotensin (1–7) may provide a strategy to limit the negative central effects of AngII.
Fetal exposure to high levels of corticosterone in a low birth weight rat model
Authors: Hennington Bettye Sue , Williamson Danielle T. , Porter Nicholas K. , Ojeda Norma B. , Intapad Suttira , McMillan Tiarra , Arguello Irene , Brown Andrew , Wilkening Stephen , Alexander Barbara T.
Published: June 2021

Fetal exposure to maternal steroids is suggested as a potential mechanism contributing to the developmental programming of adult CVD risk. 14‐day timed pregnant SD Dams were used for the RUPP while matched control SD females were subjected to sham surgeries. Corticosterone levels in Dam plasma were significantly higher in RUPP compared to E19 Control Dams. At P14, the corticosterone levels remained significantly higher in the RUPP compared to the Control Dams. Pups were sacrificed at E19, E20, and P14. E19 and E20 fetuses showed significant decrease in weight compared to the control fetuses. P14 IUGR showed significantly decreased weight compared to control pups. E19 and E20 placental weights significantly decreased in the RUPP compared to the control group. E19 and E20 placental efficiency significantly increased in the RUPP compared to control fetuses. No significant difference in plasma corticosterone between the E19, E20, and P14 IUGR compared to the Control groups. qPCR showed increased 11betaHSD2 message at E20 compared to Control placentas. Western blot analysis with kidney homogenates indicated significantly increased 11betaHSD2 in the P14 RUPP compared to Control Dams. The preliminary data suggests that while there is significant increase in plasma corticosterone in the RUPP compared to the Control Dams, the corticosterone is not crossing the placenta. The indication is that 11betaHSD2 may be part of the fetal protective mechanism; however, not the major mechanism.
Methylation of Thiols and Thioethers by Human Indolethylamine‐N Methyl Transferase
Authors: Chu Uyen , Mavlyutov Timur , Schulman Amanda , Baker Erin , Raj Rebecca , Epstein Miles , Guo Lian , Ruoho Arnold
Published: June 2021

Indolethylamine‐N MethylTransferase (INMT) is a small molecule N‐methyltransferase that transfer methyl groups from S‐Adenosyl‐L‐Methionine (SAM) to indoleamines. Methylation of tryptamine produces the hallucinogen, N,N‐dimethyltryptamine (DMT), and methylated serotonin generates bufotenin. In animals, the endogenous production of DMT has been proposed to be a regulator of the Sigma‐1 Receptor and for mood and cognition through activation of serotonin and other receptors. In contrast to the human and rabbit INMT, mouse INMT (referred to as ThioEtherMethylTransferase or TEMT) has been independently characterized and methylates thioethers, dimethyl selenide and dimethyl telluride to produce their respective “onium “ products. TEMT was suggested to be involved in the detoxification of sulfides, selenides, and tellurides. The S‐methylating activities of human and rabbit INMT have not been characterized. We evaluated the efficiency of human INMT to produce methylated sulfur containing compounds. Pure human INMT methylated both thiols and thioethers including cysteamine and 2‐(thiomethyl)ethylamine (MTEA). The Km for MTEA methylation at pH 7.2 was approximately 1.2 mM. We also compared the N‐ and S‐methylating activities of human INMT over a pH range of 6.3‐8.0 and found that acidic conditions favored S‐methylation of MTEA and alkaline conditions favored N‐methylation of tryptamine. The methylations of tryptamine and MTEA were approximately equal at pH 6.8. These data suggest a mechanism of cellular regulation of the bifunctional N‐ and S‐methylating activities of human INMT that may depend on the prevailing acidic and basic conditions of certain mammalian tissues.
You Are as Old as Your Arteries ‐ And Microvessels
Authors: Ungvari Zoltan
Published: June 2021

Nearly four centuries ago the famous English physician Thomas Sydenham (1624‐1689) commented that “a man is as old as his arteries.” Cardiovascular diseases are the indeed most common cause of death among the elderly patients in the Western world. It is becoming evident that aging results in well‐defined phenotypic changes, which render the cardiovascular system prone to disease even in the absence of traditional risk factors. The consequences of vascular aging range from myocardial infarction through erectile dysfunction to stroke. In addition to its deleterious effects on large arteries aging also results in functional and phenotypic alterations at the level of the microcirculation. There is increasing evidence that age‐related microvascular impairment plays a critical role in the pathogenesis of vascular cognitive impairment (VCI). The potential roles of microvascular mechanisms involved in VCI, including microvascular rarefaction and disruption of the blood‐brain barrier will be discussed. Also, moment‐to‐moment adjustment of cerebral blood flow via neurovascular coupling is essential for the maintenance of normal neuronal function. Increased oxidative stress that occurs with aging was shown to impair neurovascular coupling, which likely contributes to a significant age‐related decline in higher cortical function, increasing the risk for VCI. There are suggestions in the literature and new data will be presented that in aged laboratory rodents neurovascular coupling and endothelium‐dependent cerebromicrovascular dilation can be rescued, which represents a potential therapeutic target for the promotion of healthy brain aging.
Urolithin C, a Gut Microbiota Metabolite Derived from Ellagic Acid, Attenuates Triglyceride Accumulation in Human Adipocytes and Hepatoma Huh7 Cells
Authors: Kang Inhae , Tomás‐Barberán Francisco , Carlos Espín Juan , Chung Soonkyu
Published: June 2021

Urolithins (Uros) are gut microbiota‐derived metabolites from ellagitannins and ellagic acid (EA), naturally occurring polyphenols abundantly found in pomegranate, tea, some berries, and nuts. EA exerts various health benefits including TG‐lowering effects. Emerging evidence suggests that different Uros are produced in response to host's metabolic health and that each Uros may potentiate or nullify the health benefits of EA. Hitherto, little is known about the impact of individual Uros on lipid metabolism. The aim of this study is to assess potency of individual Uros in regulating lipid metabolism using human adipocyte and hepatoma cell lines. To determine the impact of Uros in adipocyte differentiation, Uro‐A, ‐B, ‐C, ‐D, and isoUro‐A (30mM) were added to differentiating human adipogenic stem cells (hASCs). Despite structural similarities, only Uro‐C was effective in reducing adipogenesis evidenced by TG accumulation, adipogenic gene and protein expressions. Consistently, Uro‐C potently inhibited de novo FA synthesis ([3H]‐acetate into [3H]‐FA) and TG esterification ([14C]‐OA into [14C]‐TG) in fully differentiated human adipocytes. Furthermore, Uro‐C, but not the other Uros, significantly attenuated TG accumulation and lipogenic gene expression in human hepatoma huh7 cells, which is comparable to EA treatment. Taken together, our results demonstrated that Uro‐C is the most biologically active gut metabolite of EA exerting potent lipid‐lowering effects both in adipocyte and hepatocytes.
Sale of micronutrient powders (MNPs) by frontline workers (FLWs) enables high reach, but low uptake limits impact on anemia and iron status: a cluster randomized study in Bangladesh
Authors: Rawat R , Saha K , Kennedy A , Ruel M , Menon P
Published: June 2021

Little is known about how different delivery platforms can be used to reach nutritionally at risk populations with MNPs. A cluster randomized design was used to examine impact, on anemia and iron deficiency (ID), of the sale of MNPs by BRAC FLWs ‐ MNP intervention, and/or enhanced nutrition interpersonal counseling (IPC) ‐ EIPC intervention. Cross‐sectional surveys at baseline (2010) and endline (2014) were conducted in children 6‐11 mo at baseline and endline, and cluster adjusted difference‐in‐difference analysis was used to estimate impact. Compared to the control group, there were no significant differences in changes in Hb (9.9 → 9.7 g/dL vs. 9.9 → 9.8 g.dL, p = n.s.) or adjusted serum ferritin (SF) conc. over time (26.9 → 21.8 ug/L vs. 24.6 → 20.2 ug/L, p = n.s.) in the MNP + EIPC group, and in the prev. of anemia (67.5 → 71.6% vs. 67.2 → 75.3%, p = n.s.) or ID (11.0 → 22.9% vs.14.7 → 28.8%, p = n.s.). In the MNP + EIPC group FLW reach was high (~90%), as was awareness of MNPs (71.9%). In this group, 50% of HHs reported ever purchasing MNPs; mean purchase in the last 6 mo was 12.1 sachets; mean consumption in the last 7 d was 1.0 sachets. Among high target children 6‐7 mo of age, only 8.3% of children were classified as regular consumers (>2 sachets in the previous 7 d). Lack of awareness of the benefits of MNPs and cost were identified as barriers to purchase and use. The lack of impact on anemia and ID appears driven by low purchase, despite high reach of the FLW network, pointing to the potential of this platform if efforts aimed at increasing demand creation and affordability are strengthened. Funding:Bill & Melinda Gates Foundation, Global Alliance for Improved Nutrition
Giardia duodenalis Depletes Goblet Cell Mucins and Degrades MUC2, Facilitating Bacterial Translocation
Authors: Amat Christina , Motta Jean‐Paul , Bhargava Amol , Chadee Kris , Buret Andre
Published: June 2021

Giardia duodenalis, a cosmopolitan diarrheagenic parasite of the small intestine, leads to post‐infectious irritable bowel syndrome (PI‐IBS) and extraintestinal complications via unclear mechanisms. The effects of Giardia on the protective mucus layers of the intestine are unknown.

Objectives

C57BL/6 (WT) and Muc2‐/‐ mice, human goblet cell line LS174T, and purified human mucin were exposed to Giardia trophozoites or Giardia's secreted product to assess the effects of Giardia on MUC2 and goblet cell mucins.

Results

Infected Muc2‐/‐ mice had higher parasitic loads and inhibition of weight gain versus WT mice. Goblet cell mucin (stained with PAS/AB and WGA) was depleted in the small intestine and colon of infected WT mice. Bacterial translocation to the liver and spleen was also increased in infected mice. Immunofluorescence staining of MUC2 was reduced in human goblet cells in vitro after exposure to Giardia trophozoites. In addition, a co‐incubation of Giardia's secreted products with purified human mucin degraded MUC2.

Conclusions

MUC2 mucin is protective in giardiasis, but Giardia is able to overcome this barrier. While Giardia colonizes the upper small intestine, it induces mucin depletion in goblet cells throughout the small intestine and colon. Mucin depletion and degradation break down the protective mucus barrier, facilitating bacterial translocation. These mechanisms may contribute to PI‐IBS. Supported by HPI NSERC CREATE, NSERC Discovery and Queen Elizabeth II Master's Scholarship
Caveolin‐1, a Mechano‐Mediator, is Expressed in the Epithelial and Stromal Cells of Mice Cervix and Increases During Pregnancy
Authors: Brunson Alicia , Ohashi Takako , Jesmin Subrina , Mowa Chishimba
Published: June 2021

Although we are aware that during pregnancy the developing fetus exerts an increasing gravitational force as it grows in size, little is known about the effect and underlying mechanism of this force on the cervix. Recently our proteomic study provided the first evidence of a presence of a cluster of mechano‐transducing molecules in the cervix, including caveolin‐1 (CAV‐1), talin‐1, vinculin, pro‐filin and others. Here, we use Confocal Immunofluorescence, Western Blot and Real Time PCR analysis to verify our earlier proteomic data by examining the cellular expression, activity (phosphorylated) and levels of CAV‐1 protein and RNA in different stages of pregnancy, non‐pregnant and postpartum (days 0, days 11, 17, postpartum 8, 24 and 48 hours) in mice. CAV‐1 was localized in the epithelial and stromal (fibroblast) cells of the cervix of pregnant mice and its expression increased with advancement in pregnancy but decreased postpartum. The expression was found to be confined to the membrane in late pregnancy and postpartum. We conclude that the growing fetus likely induces mechano‐transduction in the cervix and may influence cervical remodeling and most likely the timing of birth. Funding: Office of Student Research, Appalachian State University.
Changes in Self‐efficacy and Fruit and Vegetable Intake in the FAITH Trial
Authors: Morel L'Horset Amy , Schoenthaler Antoinette , Chaplin William , Ogedegbe Gbenga , Lancaster Kristie
Published: June 2021

Background

Despite the blood pressure (BP) lowering benefit of a diet rich in fruits and vegetables (FV), the majority of Black Americans – a high risk group for hypertension – consume fewer FV servings than whites. Self‐efficacy (SE) is associated with FV intake in diverse groups, but limited research has examined FV_SE change in interventions led by lay health advisors (LHAs). The Faith‐based Approaches in the Treatment of Hypertension (FAITH) trial evaluated the effectiveness of a lifestyle intervention delivered by LHAs on BP reduction among hypertensive Blacks in churches. This study examines the changes in FV_SE and FV intake at 6 months.

Methods: Participants were randomized into either an 11‐week therapeutic lifestyle change program and 3 subsequent monthly motivational interviewing sessions led by LHAs in the churches (MINT‐TLC), or an 11‐week health education control. Measures included the NCI FV screener and a 10‐item FV_SE scale. Changes in FV_SE score and FV intake were measured from baseline to 6 months using mixed effects regression models.

Results: In this predominantly Black (96.1%) and female (76.4%) sample with a mean age of 63.4y, systolic BP was reduced but there was no change in FV intake (p=0.37). However, there was a significant treatment x months interaction with SE; FV_SE significantly increased in the MINT‐TLC group compared to the control (p<0.001). There was also a positive association between FV_SE score and FV intake (p<0.001).

Conclusion

Although FV_SE increased and was associated with FV intake, the intervention did not increase FV intake.

Funded by NHLBI R01HL092860‐01
Akkermansia Muciniphila and Gut Microbiota Richness are Associated with Improved Metabolic Status after Calorie Restriction
Authors: Dao Maria , Everard Amandine , Aron‐Wisnewsky Judith , Sokolovska Nataliya , Verger Eric , Rizkalla Salwa , Doré Joel , Cani Patrice , Clément Karine
Published: June 2021

Populations with obesity and Type 2 Diabetes differ from healthy populations in the abundance of certain gut microbial species and microbial gene richness (MGR). Abundance of Akkermansia muciniphila (Akk) has been inversely associated to fat mass and glucose intolerance in mice. More evidence is needed in humans as well as on the impact of diet and weight loss on Akk abundance. Our aim was to evaluate the association between fecal Akk abundance and MGR, host characteristics, and their changes as a result of calorie restriction (CR) for 6 weeks followed by weight stabilization (WS) for 6 weeks in 49 overweight/obese adults. Fecal Akk, MGR, diet and bioclinical parameters were measured at baseline (BS) and after CR and WS. At BS Akk was inversely related to fasting glucose, waist‐to‐hip ratio, and adipocyte diameter in subcutaneous white adipose tissue. Subjects with higher MGR and Akk had the healthiest metabolic status, particularly in fasting glucose, triglycerides and body composition. Higher BS Akk was associated with greater improvement in insulin sensitivity and other clinical parameters after CR. Akk is associated with a healthier metabolic status and outcomes after CR in this population. Further studies are required to establish underlying mechanisms in humans. Support: ANR MICRO‐Obes, KOT‐Ceprodi, Fondation Coeur et Arteres, Metacardis HEALTH‐F4‐2012‐305312, FRS‐FNRS, ERC Starting grant 336452‐ENIGMO).
Phenolic Extracts of Cranberry Positively Modulate Endogenous Antioxidant Defenses and Signalling Pathways Against Oxidative Challenge
Authors: Khoo Christina , Martin Maria , Ramos Sonia , Mateos Raquel , Bravo Laura , Marais Johannes , Goya Luis
Published: June 2021

Oxidative stress and reactive oxygen species (ROS)‐mediated cell damage are implicated in various chronic pathologies. Emerging studies show polyphenols may act by increasing endogenous antioxidant defense potential. Cranberry has one of the highest polyphenols content among commonly consumed fruits. In this study, hepato‐protective activity of two cranberry extracts (CP1 and CP2) against oxidative challenge were screened using HepG2 cells, looking at ROS production, intracellular non‐enzymatic and enzymatic defenses by reduced glutathione concentration (GSH), glutathione peroxidase (GPx) and glutathione reductase (GR) activity and lipid peroxidation biomarker malondialdehyde (MDA). Both extracts in basal conditions did not affect cell viability but decreased ROS production and increased GPx activity, conditions that may place the cells in favorable conditions against oxidative stress. Extracts pre‐treatment of HepG2 cells for 20h significantly reduced (tert‐butylhydroperoxide (t‐BOOH)‐induced cell damage compared to untreated cells. Both extracts (5‐50ug/ml) reduced t‐BOOH‐induced increase of MDA by 20 % (CP1) and 25 % (CP2), and significantly reduced increase in GPx and GR. CP2 prevented a reduction in GSH and significantly reduced ROS production. CP1 reverted the t‐BOOH‐induced increase in p‐JNK. This study demonstrates that cranberry polyphenols may help protect liver cells against oxidative insult by modulating GSH concentration, ROS and MDA generation, antioxidant enzymes activity and cell signaling pathways. Funded by Ocean Spray Cranberries.
BPC 157 Counteracts Gastric Lesions after Bilateral Nephrectomy and Attenuates Deleterious Course in Rats
Authors: Drmic Domagoj , Sucic Mario , Zenko Sever Anita , Kolenc Danijela , Suran Jelena , Seiwerth Sven , Sikiric Predrag
Published: June 2021

Aim. Stable gastric pentadecapeptide BPC 157 is an antiulcer peptide tested in clinical trials, for inflammatory bowel disease and multiple sclerosis, effective against a variety of different ulcerogens, NSAIDs, absolute alcohol, restraint stress, insulin, alloxan, acute pancreatitis. However, BPC 157 was not tested against gastric lesions that might be induced with bilateral nephrectomy. Of note, chronic renal insuficiency might be associated with increased peptic ulcer presentation. Materials and methods. Rats were subjected to bilateral nephrectomy, sacrificed at 24h and at 48h, BPC 157 was given (10µg/kg, 10ng/kg i.p.) immediately after nephrectomy and 24h after nephrectomy. Assessment included gastric lesions, gross clinical presentation, urea, creatinine, AST, ALT, bilirubin, K, Na, Mg, Ca, inorganic phosphates in serum as described (Sikiric et al, Curr Pharm Des. 2014;20(7):1126‐35). Results. Gastric lesions were consistently attenuated (means±SD, sum of the longest lesions diameters, 1.8±0.5 mm µg (24h), 1.0±0.5 mm µg (48h) vs. 10.5±1.5 mm control (24h), 15.0±3.0 mm control (48h), *P<0.05, at least, the increased values of the urea, creatinine, AST, inorganic phosphates were lessened while K, Na, Mg, Ca, were not affected. All controls rats were edematous (scored 0‐3) (Min/Med/Max, scored 3/3/3) while BPC 157 were markedly less affected (scored 1/1/1 µg, at 24h and 48h; 1/1/1 ng, at 24h and 48h). Conclusions. Applied after nephrectomy, the deleterious course was attenuated after BPC 157 administration, and particularly, gastric lesions markedly attenuated. Supported by Grant 108‐1083570‐3635, Croatia.
Isolation and Purification of Entamoeba histolytica Alcohol Dehydrogenase 2 (EhADH2) Enzymatic Activities and Inhibition by Pyrazoline Derivatives
Authors: Leitao Joshua , Oduaran Erica , Rossi Lauren , Espinosa Avelina
Published: June 2021

The bifunctional alcohol/ aldehyde dehydrogenase enzyme in Entamoeba histolytica (EhADH2) belongs to the ADHE iron dependent family, and is essential for trophozoite growth and survival. EhADH2 catalyzes the conversion of acetyl Co‐A to acetaldehyde and the final reduction of acetaldehyde to ethanol by its separate ADH and ALDH domains respectively. The primary focus of research was to determine the effect of alternative treatments for amebiasis through inhibition by pyrazoline inhibitors on the EhADH2 enzyme. Series 1a (1,3‐diphenyl‐1‐carbamoyl‐2‐pyrazoline), series 1b (3‐diphenyl‐1‐propylcarbamoyl‐2‐pyrazoline), series 2 (1,3,4‐triphenyl‐1‐carbamoyl‐2‐pyrazoline), pyrazoline derivatives showed capabilities to inhibit EhADH2 activities. A kinetic assay was performed to calculate the Ki value for inhibitor [3‐(4‐chlorophenyl)‐1‐(bromophenylcarboxamide)‐2‐pyrazoline], inhibitor [3‐4(‐chlorophenyl)‐1‐N‐propylcarboxamide‐2‐pyrazoline], and inhibitor [3‐(4‐cholorphenyl)‐4‐phenyl‐1‐(4‐chlorophenyl carboxamide)‐2‐pyrazoline] at the rate of enzymatic affinity with the substrate acetaldehyde at NAD+ μM/min as the unit of measurement; then, the Ki value for the inhibitor was calculated using Michaelis‐Menten and Lineweaver‐Burke equations. Inhibitor [3‐(4‐chlorophenyl)‐4‐phenyl‐1‐(4‐chlorophenyl carboxamide)‐2‐pyrazoline], from series 2, demonstrated higher inhibition of EhADH2 enzymatic activities compared to series 1a and 1b. Future modifications to the basic structures of the three series shall improve the efficiency of these compounds.
Dietary high oleic canola oil supplemented with DHA oil attenuates plasma PCSK9 levels in humans
Authors: Pu Shuaihua , Rodríguez‐Pérez Celia , Ramprasath Vanu , Segura‐Carretero Antonio , Jones Peter
Published: June 2021

Proprotein convertase subtilisin/kexin type 9 (PCSK9) is a novel circulating protein, which plays an important role in regulation of cholesterol metabolism by promoting hepatic LDL receptor degradation. However, the action of dietary fat composition on PCSK9 levels remains unknown. A double‐blinded RCT, consisting of five 30‐d intervention periods, was conducted to investigate varying dietary oils on circulating PCSK9 levels in healthy humans. Treatments included 60 g/d of conventional canola oil (Canola), high oleic canola oil (CanoalOleic), high oleic canola/DHA oil blend (CanolaDHA), corn/safflower oil blend (CornSaff) and flax/safflower oil blend (FlaxSaff). PCSK9 levels using ELISA were assessed at the end of each phase. Lipid profiles (n=54) showed that CanolaDHA feeding resulted in the highest (p<0.05) serum total cholesterol (TC) and LDL‐C levels among all five treatments. CanolaDHA feeding also produced the lowest (p<0.05) plasma PCSK9 concentrations (204.5±9.6 ng/ml) compared to Canola (231.6±9.6 ng/ml) and CanolaOleic (230.2±9.6 ng/ml) diets. In CanolaDHA diet, PCSK9 levels correlated (r=0.35, p<0.01) with TC levels but not with LDL‐C levels. Results indicate that post‐diet response in PCSK9 may be altered with CanolaDHA diet. In conclusion, the elevated LDL‐C levels from a DHA oil treatment may not be deleterious as they are accompanied by a decline in PCSK9 levels. (Supported by the Canola Council of Canada and Growing Forward II program)
Variability in Plasma Free Choline and its Relation with Diet and Potential Plasma Biomarkers
Authors: Wiedeman Alejandra , Dyer Roger , Innis Sheila
Published: June 2021

Choline is an essential nutrient that is crucial for synthesis of phoshatidylcholine (PC), which is important for cell membranes and plasma lipoproteins. Choline is also involved in the synthesis of the neurotransmitter acetylcholine, and when oxidized to betaine, serves as a methyl donor. Choline is present in a variety of foods, in different forms including PC. Additionally, choline is endogenously synthesized through the sequential methylation of phosphatidylethanolamine. The current biomarker used to assess choline status is plasma free choline, usually is measured at one time only; moreover is not the only form of choline in plasma. The relation of plasma free choline with the different choline forms present in the diet and plasma is not known.

To assess intra‐individual variability of plasma free choline, and the relation with dietary intake and plasma PC and lyso‐phosphatidylcholine (LPC).

This study involved 40 adults and used a repeated measure design, and blood samples were collected in different days. Plasma was analysed for free choline using LC‐MS/MS, and for PC and LPC using HPLC. Dietary intake of choline was calculated with nutritional software using database for choline.

Within individuals, plasma free choline varied up to 83%, between different sample time points. However, comparing the mean from all participants showed a variance of only 30%. Different forms and total choline intake were not associated with plasma free choline values. The next step is to complete the analysis of plasma PC and LPC.

Studies focusing in assessing plasma free choline need to be aware of the large daily individuals' fluctuation in this biomarker.

Supported by CIHR grant and Becas‐Chile scholarship.
Effects of Probiotics on Glycemic Control and Inflammation in Type 2 Diabetes Mellitus: A Randomized, Double‐Blind, Placebo‐controlled Study
Authors: Martino Hercia , Tonucci Livia , Santos Karina , Oliveira Leandro , Ribeiro Sonia
Published: June 2021

Aims

The aim of this clinical trial was to investigate the effects of probiotics on glycemic control and inflammatory and anti‐inflammatory markers in T2D.

Methods

In a double‐blind, randomized controlled trial, 50 subjects with T2D were assigned to two groups: probiotic group, consumed 120 g/d of fermented milk containing Lactobacillus acidophilus LA5 and Bifidobacterium lactis BB12 or placebo group, consumed 120 g/d of conventional fermented milk for 6 weeks. Anthropometric measurements, body composition and fasting blood were took at baseline and after 6 weeks intervention.

Results

The study demonstrated a significant decrease in fructosamine levels (p = 0.05) and haemoglobin A1c (HbA1c) tended to be lower (p = 0.07) in probiotic group. TNF‐ α and resistin were significantly reduced (p < 0.05) in both groups after end of trial, while IL‐10 was significantly reduced (p < 0.001) and adiponectin levels tended to be lower (p = 0.07) in the placebo group. There was a significant difference in mean changes of HbA1c, between groups (p < 0.05). No significant changes from baseline were shown in IL‐6 levels.

Conclusion

Probiotic consumption improved the glycemic control and prevented a reduction in anti‐inflamatory cytokines in T2D subjects. The Fermented milk intake seems to be involved with reductions in inflammatory markers.

Research Support: Brazilian Foundation CAPES and CNPq
Aminolevulinic Acid Treatment of Pulmonary Arteries Attenuates Endothelin‐1 and Angiotensin II Elicited Increases in Mitochondrial, but not Extra‐Mitochondrial Superoxide
Authors: Patel Dhara , Alhawaj Raed , Kelly Melissa , Froogh Ghezal , Sun Dong , Wolin Michael
Published: June 2021

We recently reported that treatment of mice with the heme biosynthesis precursor δ‐aminolevulinic acid (ALA) attenuates the development of hypoxia‐induced pulmonary hypertension. In addition, we previously reported that 24‐hour organoid culture of bovine pulmonary arteries (BPAs) with ALA caused an increase in soluble guanylate cyclase (sGC) activity by generating protoporphyrin IX, a stimulator of sGC and cGMP‐mediated activation of PKG. This study examines how heme biosynthesis modulation with ALA potentially functions to prevent the effects of 24‐hour organoid culture of BPAs with the hypoxia and pulmonary hypertension mediators endothelin‐1 (ET‐1) and angiotensin II (AngII), with a focus on detecting subcellular changes in superoxide production. Organoid culture of BPAs with 10 nM ET‐1 or 0.1 µM AngII for 24‐hours caused increases in mitochondrial (MitoSox/HPLC) and extra‐mitochondrial/cytosolic (DHE/HPLC) superoxide, but ALA only attenuated the increases in mitochondrial superoxide. Since ALA also attenuates an ET‐1 elicited remodeling regulatory mechanism controlled by src kinase phosphorylation of STAT3 and its associated lowering of miR204 in organoid cultured BPA and the chronic hypoxia‐elicited depletion of miR204 in mouse lungs, the prevention of increased mitochondrial, but not extra‐mitochondrial superoxide generation could be a key factor in how ALA attenuates processes in pulmonary arteries that contribute to hypertension development.
Angiotensin II Dependent Regulation of TRPC6 Calcium Channels in the Podocytes of the STZ‐induced Type 1 Diabetic Dahl SS Rats
Authors: Ilatovskaya Daria , Palygin Oleg , Lowing Andrea , Levchenko Vladislav , Staruschenko Alexander
Published: June 2021

Injury to podocytes (terminally differentiated epithelial cells of the glomeruli) is considered a major contributor to diabetic kidney disease, when podocyte loss causes proteinuria and progressive glomerulosclerosis. Podocyte depletion may result from improper calcium handling in these cells, f.i. due to abnormal activation of the TRPC channels stimulated by Angiotensin II (Ang II). The goal of this study was to assess the ramifications of type 1 diabetes on Ang II mediated calcium influx in the podocytes. Diabetes was induced in the 6 week old Dahl salt‐sensitive rats by an injection of STZ, which resulted in weight loss, excessive urination and renal damage typical for diabetic nephropathy. During next 12 weeks rats were monitored to ensure hyperglycemia; biochemical analyses showed progressive proteinuria and high nephrin excretion, whereas excretion of Na+ and K+ remained normal. Patch‐clamp electrophysiology performed on podocytes of the isolated glomeruli showed increased basal TRPC6 channel activity (FNPo) in the STZ‐treated animals compared to controls (41% and 18% of patches, respectively), and a 5‐fold elevation of the channels open probability in response to 10 µM Ang II (the conductance (18 pS) and reversal potential of the channel (0 mV) remained unchanged). Ratiometric confocal measurements in glomeruli loaded with Fluo4/FuraRed dyes showed increased calcium influx in response to Ang II in diabetic rats compared to control animals. We can conclude from these data that Ang II‐dependent TRPC6 channels regulation plays a critical role in the glomerular filtration barrier destruction during type 1 diabetes.
Intestinal vitamin D receptor protects mice from dysbiosis via modulating JAK/STAT pathway in tumorigenesis
Authors: Wu Shaoping , Zhang Yong‐guo , Lu Rong , Xia Yinglin , Zhou David , Sun Jun
Published: June 2021

Low vitamin D receptor (VDR) expression and diminished vitamin D/VDR signaling are observed in patients with colon cancer. We reported that intestinal epithelial VDR deletion (VDRΔIEC ) leads to dysbiosis and increases susceptibility to colitis. The JAK/STAT pathway plays a critical role in intestinal and microbial homeostasis. We hypothesized intestinal VDR protects mice from dysbiosis via modulating the JAK/STAT pathway in tumorigenesis. We used an azoxymethane/Dextran sulfate sodium ‐induced cancer model in VDRΔIEC mice. VDRΔIEC mice have higher tumor numbers with tumor location shifted from distal to proximal colon. Fecal microbiota analysis showed that lacking VDR leads to bacterial profile shift from normal to susceptible carcinogenesis. We found enhanced bacterial staining in mouse tumors. These data are consisted with our findings in human tumor sample. VDR deletion decreased Jak2 at protein and mRNA levels. By CHIP assay, we identified that VDR protein bound to the Jak2 promoter, suggesting that VDR transcriptionally regulated Jak2. JAK/STAT was the upstream of the proinflammatory NF‐kappa B pathway. Furthermore, IκBα expression was significantly decreased in VDRΔIEC tumors. Our observations suggest that absence of intestinal epithelial VDR leads to dysbiosis and susceptibility to colon cancer via reducing JAK/STAT signaling and dampening inflammatory responses. Our study provides new insight into the molecular mechanism of VDR regulating the JAK/STAT pathway in bacterial‐host interaction and tumorigenesis. Our studies also indicate a new target—microbiome and VDR signaling for prevention of colon cancer.
Anatomy Education for the YouTube Generation
Authors: Barry Denis , Tierney Paul , O'Keeffe Gerard
Published: June 2021

Anatomy remains a cornerstone of medical education despite challenges that have seen a significant reduction in contact hours over recent decades; however, the rise of the YouTube Generation, or GenC, offers new possibilities for anatomy education. GenC, which consists of 80% Millennials, actively engage with social media and integrate it into their education experience. Most are willing to merge their online presence with their degree programs by engaging with course materials and sharing their knowledge freely using these platforms. This integration of social media into undergraduate learning, and the attitudes and mindset of GenC has changed traditional learning approaches and the student / teacher relationship. To gauge this, second year undergraduate medical and radiation therapy students (n = 73) were surveyed regarding their use of online social media in relation to anatomy learning. The vast majority of students had employed web‐based platforms to source information with 78% using YouTube as their primary source of anatomy‐related video clips. These findings suggest that the academic anatomy community should consider integrating social media into blended learning approaches in anatomy programs. This will ensure continued connection with the YouTube generation of students while also allowing for academic and ethical oversight regarding the use of online video clips whose provenance may not otherwise be known.