bluuebunny/crossref_metadata_2025 huggingface dataset:
Effect of administration of iron and calcium tablets on the nutritional status, physical development, and bone density of preschool children
Authors: Shin Kyung Ok , Park Hyun Suh , Chung Keun Hee
Published: June 2021

Several nutritional hazardous factors have been identified in children who prefer an unbalanced diet‐these include deficiencies of Fe, Ca, Zn, niacin, and other nutrients, all of which appear to be associated with underweight. Food intake in children and adolescents is of particular concern, because dietary status may exert a considerable effect on development, bone density, and life‐long health status. Therefore, in the present study, we administered these nutrition‐hazardous factors (Fe and Ca) to experimental groups for a period of four months. For the subjects (135 children) who continued to participate until the end of this study, we assessed increases in nutrient intake. According to our physical examination and bone density tests : The RDA average value of Energy (p<0.01), protein (p<0.003), vitamin A (p<0.04), Calcium (p<0.0001), and Zn (p<0.008) was reduced after dietary administration, and vitamin B2 was lower (123.4%) than prior to administration (135.1%). The childrens' height was not shown to differ between prior to and after dietary administration, and the childrens' weight was higher in the group treated with Fe (15mg) and Ca (232mg) than in the group treated with Fe (0.63mg) and Ca (27.5mg). The WLI value after dietary administration was higher in the Fe (15mg) and Ca (232mg)‐administered group as compared with other groups (p<0.024). However, all children were at an age of growth, and the growth rate was increased in all groups. Among the Fe and Ca administrated‐groups A(Fe 0.63mg, Ca 27.5mg), B(Fe 15mg, Ca 232mg), C(Fe 30mg, Ca 232mg), and D(Fe 15mg, Ca 462mg), the bone density was higher in the B, C, and D groups than in group A.
High sodium intake interacts with ADD1 Gly460Trp polymorphism in increasing hypertension odds among Filipino women
Authors: Lee Nanette R , Borja Judith , Duazo Paulita , Mohlke Karen , Adair Linda S
Published: June 2021

Hypertension (HTN) is a multifactorial disease influenced by environmental and genetic factors. We examined the association between high sodium (Na) intake and HTN among 1747 unrelated Filipino women (ages 35‐69 yr) and assessed whether the association is modified by a gene polymorphism (Gly460Trp of alpha adducin (ADD1)) shown to increase Na re‐absorption in the kidney. We used data gathered by the Cebu Longitudinal Health and Nutrition Survey in 2005 and defined high Na intake as: at least 4600 mg/day (200% of recommended upper limit). Mean Na consumption was 2900 mg/day, 15% were categorized as high intake, and 29% were hypertensive. Adjusting for age, BMI, overweight duration and other confounders, logistic regression analyses showed that high Na intake increased HTN odds, especially among women homozygous for the 460Trp allele (19%). Women with lower Na intake carrying one or more copies of the 460Gly allele (reference) had the lowest odds of HTN: odds ratio (OR, 95% CI) for women with high Na intake carrying the 460Gly allele was 1.2 (0.8, 1.6) while that for women with high Na intake and homozygous for 460Trp was 2.9 (1.4, 6.0). The association among Na intake, ADD1 Gly460Trp, and hypertension was further modified by overweight duration. With relatively high frequency of the 460Trp allele compared to Caucasians, efforts to reduce Na intake should be strengthened.

1R03TW008133‐01, 1 R01 HL085144‐01, R01 DK078150
Determinants of anemia at 1 mo postpartum in Mexican women supplemented during pregnancy
Authors: Dominguez‐Islas Clara Penelope , Garcia‐Guerra Armando , Mejia‐Rodríguez Fabiola , Neufeld Lynnette Marie
Published: June 2021

The objective of this analysis was to explore determinants of anemia at 1 mo postpartum (PP) in women supplemented with multiple micronutrients during pregnancy. Data came from a cluster randomized trial where pregnant women received a fortified beverage (FB), tablets (TB) or Sprinkles (SK). Supplements had identical micronutrient content with 15 mg of iron and were delivered daily in the home. Appropriate statistical analyses for cluster randomized trials were used. Mean duration of pregnancy at recruitment was 20.7±5.8 wks, hemoglobin concentration (Hb) at recruitment was 12.1±1.4 g/dL (21.0% anemic, Hb<11 g/dL), and the mean of doses consumed from recruitment to 1 mo post‐partum was 113.8±38.5. At 1 mo PP, 25.9% of women were anemic (Hb<12 g/dL). Factors that were found to be associated with the probability of being anemic include the time since birth of previous child (OR 0.98, p<0.01), Hb at recruitment (OR 0.52, p<0.01), increment of Hb from recruitment to 37 wks pregnancy (OR 0.62, p<0.01), caesarean delivery (OR 1.6, p=0.08) and type of supplement consumed (OR 1.83, p=0.02 for FB and OR 1.66, p=0.06 for SY, respect to TB). A model with the same independent variables explained 22% of variance for Hb concentration at 1 mo PP. In conclusion, despite reasonable compliance with supplementation, the prevalence of anemia at 1 mo PP was high and associated with a number of factors, including the type of supplement consumed.
Maternal HIV status and motor milestone acquisition among Ghanaian infants
Authors: Lartey Anna , Marquis Grace S , Perez‐Escamilla Rafael , Mazur Robert , Sellen Daniel
Published: June 2021

As part of the RIING project (Research to Improve Infant Nutrition and Growth) we examined infants' motor milestone (MM) acquisition in relation to Ghanaian mothers' HIV status. Mothers' prenatally diagnosed as HIV positive (n=104), HIV negative (n=132) and HIV status unknown (n=129) were followed up twice weekly at home for MM assessment from 4 to 12 months. At each home visit the following were assessed: i) sitting without support, ii) hands and knees crawling, iii) standing with assistance, iv) walking with assistance, v) standing alone, and vi) walking alone. The mean age (days) for MM achievement was not significantly different among children of the three groups of mothers (all groups combined: sitting without support 170.7± 24.5 d, hands and knees crawling 243.0± 34.9 d, standing with assistance 261.0± 38.9 d, walking with assistance 295.3± 46.8 d, standing alone 329.0± 36.7 d, and walking alone 350.3± 35.1 d) However, by 12 months 15.4 % of the HIV positive, 31.8% of the HIV negative, and 10.1% of the HIV unknown mothers' children were walking alone (p<0.05). The results suggest possible influence of maternal HIV status on child MM acquisition.

Funded by NIH Grant HD436020
Maternal postpartum depression modifies the association between maternal HIV infection and infant diarrhea in Ghana's Eastern region
Authors: Marquis Grace S , Okronipa Harriet , Lartey Anna , Brakohiapa Lucy , Perez‐Escamilla Rafael , Mazur Robert E
Published: June 2021

Diarrhea remains a major public health concern in low‐income countries. We examined the incidence and determinants of diarrhea among infants of HIV positive (HIV‐P), HIV negative (HIV‐N), and unknown HIV status (HIV‐U) women. Pregnant women (n=492) were recruited from three antenatal clinics after voluntary counselling and testing and followed from 0 to 12 mo after delivery. The incidence of diarrhea in the first 12 mo of life was 0.6 episodes per 100 d of observation; there was no group difference by maternal HIV‐status. Infants of HIV‐U compared to HIV‐P and HIV‐N women tended to have more days ill with diarrhea between 0‐3 mo (P=0.08), 3‐6 mo (P=0.06), and 0‐12 mo (P=0.05). HIV‐P compared to HIV‐N and HIV‐U women tended to report symptoms of post‐partum depression (PPD) at 0 mo (P=0.10). Infants whose mothers showed symptoms of PPD at 0 mo had over twice as many days ill with diarrhea (P<0.001) and more diarrheal episodes (P=0.002) between 0‐3 mo. Multivariate analyses showed that between 0‐3 mo of age, there was a significant interaction (P=0.02) between maternal HIV status and PPD. Symptoms of PPD at 0 mo increased the risk of 0‐3 mo diarrhea for HIV‐P (OR=6.2; 95% CI: 2.7, 14.3) and HIV‐U (OR=3.5; 95% CI: 1.9, 6.7) women, but not for HIV‐N women. Support systems in health care facilities should include program components that address PPD, especially in HIV‐affected communities. Funded by NICHD HD43260
Metabolic Syndrome in HIV+ Asymptomatic Adults in Botswana, Africa
Authors: Sales Sabrina , Campa Adriana , Makhema Joe , Dusara Priti , Tsalaile Lesedi , Burns Patricia J , Baum Marianna K , Marlink Richard
Published: June 2021

Objective

To determine the prevalence of metabolic syndrome (MS) in a cohort of asymptomatic and antiretrovirally naïve HIV+ adults in Botswana, and its association with other risk factors for cardiovascular disease (CVD).

Methods

After consenting, socio‐demographic and medical questionnaires were completed, blood pressure and anthropometries taken. Fasting blood was drawn for total blood cholesterol, HDL cholesterol, triglycerides, and glucose. MS was defined according to NCEP ATP III guidelines.

Results

Of the 402 participants, 69.4% were female and the mean age was 33.7±8.1 years. The prevalence of MS in this cohort was 7.7% (n=31), 32.8% were overweight (BMI >25 kg/m2), 11.6% were obese (BMI >30 kg/m2), 21.9% had hypertenstion, 31.9% reported alcohol use and 19.5% tobacco use. Only 5.1% had high cholesterol and 14.3% high LDL levels. Risk factors significantly associated with MS were age (OR=1.10, 95% CI: 1.04, 1.16, p<0.01) and BMI (OR=1.26, 95% CI: 1.16, 1.36, p<0.01) after adjusting for age and gender.

Conclusions

The prevalence of MS in this cohort is similar to other sub‐Saharan African HIV+ cohorts, but low compared to that in developed countries. Preventive interventions should be implemented to reduce the future impact of MS on HIV‐related CVD risk, especially when these patients start their antiretroviral treatment.

Funded by NIDA
Contribution of tea to iron status in a rural female population
Authors: Osowski Jane Marie , Specker Bonny
Published: June 2021

Objective

To investigate the association between tea consumption and iron status in a rural South Dakota female population.

Methods

The study design was cross‐sectional. This study examined whether there is a correlation between tea consumption and iron status. A convenience sample of 136 adolescent girls and women living on Hutterite colonies in rural South Dakota participated in the study. The main outcome measures included hemoglobin (Hgb), expressed as mg/dL, the amount of tea consumed, expressed in cups/meal and the amount of dietary iron consumed, expressed as mg/day. The sample was divided into two groups depending on if they consumed tea at mealtime or did not consume tea at mealtime. Differences between group means were tested using Students t test and chi‐square analysis. Multiple regression was used to assess the relationship between Hgb levels and tea consumption controlling for variables with relevance to the main outcome measure.

Results

The difference in mean Hgb concentrations between groups did not differ. The mean intake of tea at mealtime was 2 ± 1 cups. The amount of black tea consumed at mealtime was significantly more than herbal or green tea (p = 0.05). The difference in the amount of iron consumed per day between groups was not significant (p = .11). No relationship was found between hemoglobin and amount of tea consumed while controlling for variables that influence hemoglobin.

Conclusions

In a seemingly healthy rural female population consuming tea in the amounts that were observed with a whole meal does not have an effect on hemoglobin levels.
A comparison of zinc metabolism and status among young and older Korean women
Authors: Kim Jihye , Paik Hee Young , Joung Hyojee , Woodhouse Leslie R , King Janet C
Published: June 2021

This study was done to determine the effect of age on zinc metabolism and status among healthy Korean women. Zinc measures were assessed in 8 young and 7 elderly women. Oral and intravenous tracers enriched in 67Zn and 70Zn, were administered. Multiple plasma and 24‐h urines were collected for 5 d with complete fecal samples for 9 d after isotope administration. In the young women, additional plasma samples were collected to determine zinc kinetics. The size of the Exchangeable Zinc Pool (EZP) was estimated by Miller's method. Plasma zinc was higher in older women than younger women (p<0.05). EZP, EZP/kg, and urinary zinc tended to be higher in older women than younger women. Fractional and total zinc absorption and endogenous fecal zinc losses did not differ between young and older women. The zinc kinetics of the Korean and American women showed no differences in plasma or EZP parameters. However, total absorbed zinc and zinc flux to slowly turning over tissues (Q7) was significantly lower in Korean women than that of Americans (p < 0.01). The zinc status of older Korean women, as measured by plasma zinc and EZP was higher than that of the younger women although there were no changes in zinc absorption or endogenous fecal excretion. The young Korean women absorbed less zinc and transferred less zinc to Q7 than American women suggesting the total body zinc content of the Korean women is lower than that of their American counterparts.
Molecular Modeling Studies: Potential Binding of Soy Protein 7S Globulin Subunits to Human LDL Receptor
Authors: Tafti Ashkan , Hecht David , Beuhler Lukas , Bakhiet Raga
Published: June 2021

Soybean 7S Globulin subunits have been associated with a decrease in cholesterol both in in vitro and in vivo studies1. One possible mechanism could be binding of the protein or its digested fragments to the LDL receptor. In order to explore the feasibility of this hypothesis, molecular modeling experiments were performed using homology modeling software (MOE, Chemical Computing Group, Inc.) and protein‐protein docking software (FTDock, RPScore & Multidock from Biomolecular Modeling Laboratory, Imperial Cancer Research Fund). In these studies, a homology model was generated for the 7S globulin based on the x‐ray crystal structure of a hydrolase inhibitor from wheat (1T6E.pdb). The LDL receptor binding site was also obtained from the Protein Data Bank (IN7D.pdb). From these docking studies it was observed that the homology model bound to several sites of the extra‐cellular region of the LDL receptor. Possible interactions of 7S Globulin peptide fragments with cyclophilin were modeled and were identified as definite interactions or associations.
A panel of biomarkers for esophageal squamous cell carcinoma
Authors: Kashyap Manoj K , Marimuthu Arivusudar , Kishore C. J. H. , Peri Suraj , Ranganathan Prathibha , Sanjeeviah Ravinder C. , Mahmood Riaz , Kumar K. V. Veerendra , Vijayakumar M. , Montgomery Elizabeth , Kumar Rekha V. , Pandey Akhilesh
Published: June 2021

We have carried out mRNA profiling of 20 esophageal squamous cell carcinoma (ESCC) using whole genome DNA microarrays for identification of novel biomarkers. We chose several novel candidates including periostin (POSTN), lumican (LUM), four and a half LIM domains 1 (FHL1), collagen triple helix repeat containing 1 (CTHRC1) and PTRF interacting protein alpha 1 (PPFIA1) to carry out immunohistochemistry‐based validation on a larger sample set using tissue microarrays. Overexpression of periostin was observed in 139/140 (99%) of cases while. lumican was overexpressed in 141/141(100%) of cases and expression of FHL1 was found to be decreased in 132/145 (91%) of cases. These markers could be further tested in sera of patients as candidates for early detection of this cancer.

This work was supported by an Independent Senior Research Fellowship to MKK from the Indian Council of Medical Research, New Delhi, India.
A potential role for ajulemic acid in treatment of the Ewing family of tumors
Authors: Daly Sean W. , Sims John R. , Evans Emily A. , Prescott Amber E. , Tull Cameron A. , Suva Larry J. , Hensley Lori L.
Published: June 2021

Ewing's Sarcoma is a bone cancer that primarily affects children and teenagers. With a five‐year survival rate of approximately 30%, improved treatment options are needed. Our research has focused on the ability of the synthetic cannabinoid, ajulemic acid, to induce death in Ewing's sarcoma cells in vitro. We find ajulemic acid induces apoptosis in Ewing's sarcoma cells and several other tumor types in the Ewing family of tumors. In order to test the efficacy of ajulemic acid in vivo, we developed Ewing's sarcoma cells that express a firefly luciferase expression system as a handle by which to monitor tumor growth and drug response in living mice. SK‐ES Ewing's sarcoma cells were transiently transfected with firefly luciferase and then injected into the tibiae of nude mice. Tumors were allowed to develop and the growth of the bioluminescent tumors tracked using in vivo imaging techniques. The effect of different doses of ajulemic acid (and other potential therapies) can be used in these mice and tumor responses in individual mice can be monitored and measured. We hypothesize that these experiments provide the rationale for the development of improved therapies for this devastating cancer. The project described was supported by NIH Grant Number P20 RR‐16460 from the IDeA Networks of Biomedical Research Excellence (INBRE) Program of the National Center for Research Resources.
Tamoxifen inhibits the proliferation and survival of malignant peripheral nerve sheath tumor cells via an estrogen receptor‐independent mechanism
Authors: Eckert Jenell Marie , Byer Stephanie , Prasain Jeevan , Carroll Steven
Published: June 2021

No effective therapies are available to patients with malignant peripheral nerve sheath tumors (MPNSTs), the most common malignancy associated with neurofibromatosis type 1 (NF1). We have found that 4‐hydroxy‐tamoxifen rapidly induces MPNST cell death at concentrations similar to those promoting breast carcinoma apoptosis while lower concentrations inhibit MPNST mitogenesis. Although MPNSTs and MPNST cell lines express estrogen receptor β(ERβ), G‐protein coupled estrogen receptor‐1 (GPER) and estrogen biosynthetic enzymes, mass spectrometric studies shows no evidence that MPNST cells secrete 17β‐estradiol. Further, exogenous 17β‐estradiol, alone or in combination with forskolin, did not stimulate MPNST mitogenesis and ablation of ERβ and GPER expression by RNA interference likewise had no effect. Tamoxifen also acts via estrogen receptor‐independent means, including inhibition of calmodulin and protein kinase C (PKC) signaling. While the broad spectrum PKC inhibitor GF109203X had no effect, calmodulin inhibitors (trifluoperazine, W‐7) recapitulated 4‐hydroxy‐tamoxifen effects on MPNST cells. We conclude that 4‐hydroxy‐tamoxifen inhibits MPNST cell proliferation and survival via an estrogen receptor‐independent mechanism that may involve calmodulin. These observations suggest that tamoxifen may be effective for the treatment of MPNSTs. Supported by NIH grants NS048353, CA122804.
MAb C11C1, a monoclonal antibody to cleaved high molecular weight kininogen (HK) inhibits murine tumor metastasis in syngeneic hosts
Authors: Khan Sabina T , Pixley Robin A , Liu Yuchuan , Gordon Bridgette , Huang Yajue , Agelan Alexis , Achary Mohan , Colman Robert W
Published: June 2021

Metastasis of malignant tumors is a major cause of morbidity and mortality. We previously demonstrated that C11C1 has the ability to reduce tumor growth in murine multiple myeloma in mice and human colon cancer in nude mice. Normal mice treated with C11C1 did not show any gross or histological evidence of organ damage or hemorrhage. We now evaluate the antimetastatic effect of C11C1 on C57BL/6 mouse lung metastatic model using B16F10 melanoma cells. In two separate studies, tail veins of mice were injected with melanoma cells. Each study included two groups of mice. One group received C11C1 and the other received saline (control) intraperitoneally. In one study all the mice were euthanized at 28 days and in the second 52 days, to allow a longer time period to develop metastases. In the 28 day control group, 6 of 10 mice had detectable metastatic pulmonary nodules. In the 52 day control group 5 of 9 mice had metastases. There was no relation of the incidence of metastases to the duration of the study. The metastatic nodules stained positive with an antibody against S‐100 protein, a tumor antigen which is present in B16 F10 cells. In the C11C1 groups, (Fisher Exact test) 0 of 10 mice (p=0.017) and 0 of 13 mice (p=0.005) showed no metastatic foci in their lungs. These studies demonstrate that a monoclonal antibody to HK has a therapeutic potential with minimal side effects in the treatment of metastatic cancer. NIH 2T32HL007777 & 5RO1CA83121.
APE1 protein levels are increased in PIN1 depleted murine mammary tumors
Authors: Stevens Rachel L. , Schneider Ryan A. , Liu Tongzheng , Keshvara Lakhu , Hoyt Dale G.
Published: June 2021

APE1 and several other Base Excision Repair (BER) proteins are known to be phosphorylated, which may regulate their activity and expression. PIN1 is a proline isomerase that acts on proteins phosphorylated at serine/threonine‐proline motifs. Its impact on conformation can affect protein function and turnover. Here we investigated the effect of PIN1 depletion on APE1 expression in mammary tumors. EMT6 murine mammary adenocarcinoma cells lacking PIN1 were generated by stable knockdown with small hairpin RNA. Parallel control cells were established using a mutant sequence. Tumors were established by injecting cells in the 4th mammary fat pad of female BALB/c mice. Tumors lacking PIN1 grew faster and had significantly higher APE1 levels than the corresponding slower growing control tumors. To determine if APE1 expression was attributable to tumor size or PIN1 deficiency, knockdown and control tumors were harvested at different times and tumors of similar sizes were compared. PIN1 knockdown tumors again had significantly higher levels of APE1 than control tumors of similar size harvested slightly later. These results suggest that suppression of PIN1 increases APE1 synthesis or reduces its degradation in developing tumors in vivo. PIN1 may regulate the sensitivity of tumors to DNA damaging agents by its effect on this BER enzyme.
Murine mammary tumor growth is blunted in dystrophin‐deficient mdx mice
Authors: Meaney Mary Pat , Grange Robert W. , Ju Young H.
Published: June 2021

Breast cancer (BC) is the second leading cause of cancer death in women in the US. Changes to the dystrophin glycoprotein complex (DGC), a multi‐protein structure that likely plays mechanical and signaling roles, have been reported in BC cells. For example, expression of α‐ and β‐dystroglycan appears to be inversely related to tumor stage. However, systemic absence of the DGC, as seen in the mdx mouse model of Duchenne muscular dystrophy, does not appear to induce spontaneous mammary tumor formation, indicating that presence of the DGC may be necessary for breast cancer initiation. We therefore hypothesized that breast tumor growth would be altered in mdx mice compared to wild type (C57BL/6) mice. We injected E0771 murine mammary adenocarcinoma cells into mdx and C57BL/6 mice. After 3 weeks of tumor growth, blood, skeletal muscles and tumors were collected and analyzed. Growth of E0771 tumors and serum content of migration and invasion chemokine markers, RANTES and MCP‐1, was dramatically blunted in mdx mice. The dystrophin protein was not detectable in E0771 cells or tumors, suggesting that its expression, like that of other DGC components, may be suppressed or altered in this cancer cell line. The exact mechanism(s) of tumor inhibition in mdx mice is not presently known; however, our results suggest a mechanism for suppressing BC growth and progression may depend on the absence of one or more proteins in the DGC.
Activation of cardiac fibroblast phenotypes following myocardial ischemia‐reperfusion injury in transgenic mouse models
Authors: Ambler S. Kelly , Brown R. Dale , Long Carlin S.
Published: June 2021

Following myocardial injury, cardiac fibroblasts are believed to play important roles in inflammatory and wound healing processes. We hypothesized that activation of cardiac fibroblasts and myofibroblasts in response to ischemia‐reperfusion (I/R) injury could be detected with transgenic mouse strains expressing fibroblast phenotype‐sensitive promoters linked to expression of fluorescent reporter proteins (Collagen I promoter:Enhanced Green Fluorescent Protein, Coll I:EGFP or Smooth Muscle α‐Actin promoter:Red Fluorescent Protein, SMA:RFP). Activation of cardiac fibroblasts was monitored by reporter fluorescence in frozen tissue sections and by FACS analysis of enzymatically dispersed fibroblast populations. Frozen sections from hearts of reporter mice sacrificed 4 days after I/R injury induced by transient coronary artery (LAD) occlusion showed robust enhancement in cells expressing CollI:EGFP and modest increases in SMA:RFP fluorescence. Fibroblast activation was localized to the infarct zone, and was not observed in control non‐injured mice. FACS analysis of cardiac fibroblasts isolated after I/R injury showed elevations of fluorescent reporter expression (EGFP>RFP). The respective fibroblast phenotypes were maintained under culture conditions. In conclusion, fibroblast phenotype‐sensitive reporters show activation of fibroblasts in the injured myocardium in vivo.
Diabetes‐derived muscle atrophy is associated with cellular autophagy, and potential beneficial effects of interventional physical exercise on it
Authors: Lee Youngjeon , Won Jinyoung , Kang Jae seon , Hong Yonggeun
Published: June 2021

Diabetes accompanied with cellular stress, such as functional decrease of glucose uptake, hyperglycemic oxidative stress, and damaged mitochondria. Muscle atrophy occurs when the degradation rate is higher than the synthesis rate during diabetes. Here, we hypothesized that diabetes induces skeletal muscle autophagy, however, if this increased autophagy becomes chronic, autophagy‐mediated cell death occurs that could then contribute to skeletal muscle cell loss. Sprague‐Dawley rats were injected with STZ(60 mg/kg of body weight) and after 30 days, the soleus(SOL), plantaris(PL), red gastrocnemius(RG), and white gastrocnemius(WG) muscles were assessed by immunohistochemical and western blot analysis. LC3 expression, as an autophagosomal marker was detected in all muscles examined, but was reduced in RG muscles. The content of both beclin‐1 and caspase were elevated in WG and PL muscles. All muscles from diabetic animals demonstrated reduced mass when compared to controls, but only the gastrocnemius demonstrated atrophy as reflected by a reduced muscle‐to‐body mass ratio. In addition, a group of diabetic rats submitted to daily swimming for 4 wks were compared with sedentary controls, resulting in reducing autophagy in muscles. In conclusion, these results indicate that diabetes may alter autophagic activity in a muscle‐specific manner and therapeutic exercise may modify or reverse skeletal muscle abnormalities with reducing autophagy. Keywords: Autophagy, diabetes, skeletal muscle, atrophy, LC3. Supported by KRF‐E00344, BioGreen 21 Program (#20070401034006) from Rural Development Administration, South Korea.
Molecular evidence for the involvement of calcium sensitization in serotonin‐induced cerebrovascular constriction
Authors: El‐Yazbi Ahmed , Johnson Rosalyn P. , Takeya Kosuke , Walsh Emma J. , Walsh Michael P. , Cole William C.
Published: June 2021

The mechanisms contributing to serotonin (5‐HT)‐induced vasoconstriction of cerebral arteries are poorly characterized. This is important as abnormalities in control of cerebral blood flow by 5‐HT are implicated in subarachnoid hemorrhage and may be involved in migraine. Force generation by vascular smooth muscle cells depends on the balance of myosin light chain kinase (MLCK) and phosphatase (MLCP) activities that determine the level of myosin light chain (LC20) phosphorylation. Agonists that activate Rho kinase (ROK) and protein kinase C (PKC) can inhibit MLCP by phosphorylation of myosin light chain targeting subunit 1 (MYPT1) and CPI‐17, respectively. This inhibition of MLCP leads to increased phospho‐LC20 and force generation at constant [Ca2+]i, i.e. Ca2+ sensitization. Here, we have quantified LC20, MYPT1 and CPI‐17 phosphorylation in protein samples derived from pressurized segments of rat middle cerebral arteries using a novel, highly sensitive western blotting method. 5‐HT induced constriction and increased phospho‐LC20; both were reduced by ROK (H1152, 0.3 µM) or PKC (GF109203, 3 µM) inhibition. Phospho‐MYPT1 level was increased by 5‐HT, but not phospho‐CPI‐17. Our data indicate for the first time that Ca2+ sensitization via ROK‐mediated phosphorylation of MYPT1 resulting in enhanced LC20phosphorylation and force generation may contribute to 5‐HT‐induced cerebral vasoconstriction.
Inhibition of RhoA Kinase but not Protein Kinase C can potentiate Blebbistatin‐induced relaxation of smooth muscle contraction.
Authors: Eddinger Thomas J , Ratz Paul H
Published: June 2021

Smooth muscle (SM) contraction involves numerous receptors and second messengers in a complex spatiotemporal pattern of changes that regulate force production and maintenance. RhoA kinase (ROCK) and protein kinase C (PKC) are two primary participants in this complex regulatory scheme. Blebbistatin (a cell‐permeable selective and specific small molecule inhibitor of myosin II) has been reported to inhibit Pi release and thereby prevent strong binding of cross‐bridges. We found that during a K+‐induced contraction of rabbit circular fundic strips, blebbistatin inhibited the late, slow maintained force to a greater extent than the early, fast force. Pharmacological inhibition of ROCK (Y27632, H1152) had no effect on the early, fast contractile response but significantly inhibited the late, slow contractile response. In contrast, inhibition of PKC (GF 109203x) inhibited the early, fast and late, slow contractile responses. Combined blockade of ROCK and myosin II gave a greater inhibition of the late, slow contraction than blockade of ROCK or myosin II alone. This was not true for the combined blockade of PKC and myosin II. More selective inhibitors of specific PKC isoforms did not show an additive effect of specific PKC isotypes with blebbistatin in inhibiting the late slow contraction. Partial resistance to the inhibitory effect of blebbistatin suggests that the early, fast phase of SM contraction involves unique spatiotemporal regulation of SM myosin. The potentiation by ROCK inhibitors of the inhibitory effect of blebbistatin on the late, slow contractile response suggests that ROCK may regulate force‐maintenance, in part, by a mechanism supplemental to cross‐bridge activation.
Bronchial oxidative stress in equine recurrent airway obstruction
Authors: Venugopal Changaram , Mariappan Nithya , Holmes Earnestine , Francis Joseph , Eades Susan
Published: June 2021

Oxidative stress is an important characteristic of hyperactive airway diseases such as recurrent airway obstruction (RAO) in horses. The primary source of oxidative stress in RAO is from the elevated amounts of reactive oxygen species (ROS) liberated from the inflammatory cells. ROS alters signal transduction mechanisms, activate redox‐sensitive transcription factors and regulate pro‐inflammatory gene expressions. Thus, ROS levels have important consequences for pathogenesis, severity and therapeutic management of RAO. The involvement of Tumor necrosis factor‐alpha (TNF alpha) and NFkB are well established in inflammation. Therefore, the objective of the study was to determine oxidative stress in pulmonary bronchial rings of RAO‐affected and unaffected horses and to test the effectiveness of TNF alpha blockade and NFKB to reduce production of ROS. To block TNF‐alpha and NFKB we incubated the rings with Pentoxifylline and PDTC (Pyrrolidinedithiocarbamate), respectively. Bronchial rings were collected from right diaphragmatic lung lobe of RAO‐affected and unaffected horses and the ROS released by the tissues were measured in control tissues and those incubated with the antagonists, using Electron Paramagnetic Resonance (EPR). The results showed that total ROS was greater in RAO‐affected than the unaffected horses. In addition, incubation of the rings with Pentoxifylline and PDTC reduced the ROS level in both groups of horse rings. The study is suggestive of therapeutic value for these agents in RAO‐affected horses to reduce oxidative stress. Supported by LSU‐SVM Equine Health Studies Program.
Upregulation Of Cardiac NOS Due To Endotoxemia And Vagal Overactivity Contributes To The Hypotensive Effect Of Chronic Ethanol In Female Rats
Authors: El‐Mas Mahmoud M , Fan Ming , Abdel‐Rahman Abdel A
Published: June 2021

We have previously shown that chronic ethanol causes sustained hypotension in female rats. Here, we investigated: (i) roles of cardiac contractility and autonomic activity in ethanol‐induced hypotension, and (ii) whether endotoxemia‐induced upregulation of cardiac NOS underlies hemodynamic effects of ethanol. Telemetric monitoring of blood pressure (BP), heart rate (HR), and +dP/dtmax (index of myocardial contractility) was performed in female rats receiving liquid diet with or without ethanol (5% w/v, 13 weeks). Cardiovascular autonomic control was assessed by frequency analysis of interbeat intervals (IBI) and systolic BP (SBP). Compared with control rats, ethanol caused sustained reductions in dP/dtmax and BP. Spectral analysis of IBI revealed increases in high‐frequency band (IBIHF, 0.75‐3 Hz) and decreases in IBILF/HF ratio in ethanol‐fed rats, suggesting increased cardiac parasympathetic dominance. Vascular sympathetic tone was not affected by ethanol because SBP spectral bands and plasma norepinephrine remained unchanged. Myocardial iNOS and eNOS, but not nNOS, expressions and plasma endotoxin and nitrite/nitrate were increased by ethanol.

Conclusion

upregulation of cardiac iNOS and eNOS, due to endotoxemia and/or vagal overactivity, might account for reductions in dP/dtmax and hypotension caused by chronic ethanol in female rats.

Supported by Grant R01 AA014441 from NIAAA.
Inhibition of Nitric Oxide (NO) Alone Does Not Lead to the Development of Pulmonary Arterial Hypertension (PAH) in Rats.
Authors: Sun Xiaowei , Ku David D.
Published: June 2021

PAH is a deadly lung disorder that often leads to right ventricular hypertrophy (RVH) and heart failure, but the exact role of NO is not well understood. We found that an acute bolus injection of 10 mg/kg L‐NAME or chronic (1 to 2 weeks) treatment in rats resulted in a sustained systemic hypertension, while the pulmonary pressure (PP) was surprisingly unchanged. Intravenous administration of bolus phenylephrine (PE, an α receptor agonist) also resulted in a marked increase in the systemic pressure, but not PP. However, following L‐NAME treatment or induction of monocrotaline (MCT)‐induced PAH this same PE resulted in a marked increase in PulP response, suggesting an important NO modulatory role on basal pulmonary vasoreactivity. Indeed, we found that L‐NAME pretreatment markedly accelerated the development of MCT‐induced PAH and RVH in the rats. Taken together, these results demonstrate that pulmonary endothelial NO production plays an important modulatory role on pulmonary circulation and reactivity, but not the most dominating role as it is in the systemic circulation. Prior inhibition of pulmonary endothelial NO production could accelerate the development of PAH, RVH and heart failure. Further studies using NO and its donor to prevent and treat the deadly PAH disease and preservation of RV function are currently under investigation.
Melatonin inhibits NO‐induced cGMP accumulation in porcine coronary arteries
Authors: Shukla Praveen , O'Rourke Stephen
Published: June 2021

Recent studies in our laboratory indicate that melatonin modulates vascular tone by interfering with nitric oxide (NO) signaling in coronary arteries. Here we tested the hypothesis that melatonin impairs NO‐induced vasorelaxation by inhibiting the NO‐induced increase in cGMP levels in coronary arteries. In organ bath studies, sodium nitroprusside (SNP) (10−9‐10−5 M) caused relaxation of isolated porcine coronary artery rings. Incubation of the tissues with melatonin (10−7 M) caused a rightward shift in the SNP concentration‐response curve. In the presence of the phosphodiesterase inhibitor, zaprinast (10−5 M), the inhibitory effect of melatonin on SNP‐induced vasorelaxation was abolished. Melatonin (10−7 M) had no effect on basal cGMP levels but markedly inhibited the SNP‐induced increase in cGMP. The effect of melatonin on cGMP accumulation was significantly diminished by incubation of the tissues with either zaprinast (10−5 M) or the MT2‐receptor antagonist, 4P‐PDOT (10−7 M). The expression of MT2‐receptors in coronary arteries was confirmed by immunoblot analysis. Taken together, these results suggest that the inhibitory effect of melatonin on NO signaling in coronary arteries is most likely due to impaired cGMP accumulation in response to NO. The inhibitory effect of melatonin is mediated by MT2‐receptors, perhaps via a mechanism that is coupled to increased phosphodiesterase activity.
Effect of quercetin on reactivity of sub‐chronic cadmium‐exposed rat aorta
Authors: Trakranrungsie Nopamart , Yatmark Paranee , Akkhawattanangkul Yada
Published: June 2021

The effect of quercetin, a potent radical scavenging flavonoid known to associate with reduced risk of cardiovascular disease, on reactivity of rat aorta was investigated in a sub‐chronic, orally cadmium‐exposed model. Isometric vascular measurement was employed to access vascular responses to vasoactive substances: acetylcholine (ACh), bradykinin (BK), angiotensin II (AT II) and endothelin‐1 (ET‐1). Cadmium (Cd) exposure, at 10, 30 and 100 ppm, via drinking water for 8 weeks did not cause an alteration in reactivity to ACh, but reduced relaxation response to BK in rats receiving 30 ppm Cd. Co‐treatment with 50 mg/kg quercetin could significantly recover vascular BK responses. Exposure to Cd appeared to have dual effect on aortic response to AT II. At 10 ppm Cd, contraction induced by AT II was partially impaired, while at higher dose (30 ppm), the response was potentiated, particularly at 10−10 M AT II. Although aortic rings from rats receiving quercetin alone demonstrated the marked loss in AT II response, in Cd‐treated rats, quercetin could restore the overall vascular reactivity. In addition, reactivity of aorta to ET‐1 was notably reduced by Cd, of which the effect could be reversed by quercetin. Results also showed that exposure to Cd increased levels of serum GGT and Cd accumulation in kidneys and liver in a dose‐dependent manner. Quercetin treatment reduced Cd burden in kidney only at low dose of Cd administration.
Redefining Agonist Selectivity at Vascular α1‐ and α2‐Adrenoceptors
Authors: Crassous Pierre‐Antoine , Flavahan Sheila , Flavahan Nicholas
Published: June 2021

Phenylephrine is considered a selective α1‐adrenoceptor (α1‐AR) agonist and is widely used to characterize these receptors. We observed that the selective α2‐AR antagonist rauwolscine reversed phenylephrine‐induced constriction in isolated mice tail arteries. Experiments were therefore performed to assess the selectivity of phenylephrine and other α‐AR agonists in this blood vessel. Concentration‐effect curves to the α‐AR agonists phenylephrine, cirazoline or norepinephrine were not affected by rauwolscine (3x10‐8M). The selective α1‐AR antagonist prazosin (10‐7M) caused non‐parallel shifts in concentration‐effect curves to these agonists being more potent at high compared to low levels of constriction. After prazosin, rauwolscine (3x10‐8M) inhibited concentration‐effect curves more at low compared to high levels of constriction. Indeed, the combination of prazosin plus rauwolscine caused parallel shifts in the concentration‐effect curves to these agonists. The α2‐AR agonist UK14304 caused a low‐maximum constriction compared to the other α‐AR agonists. Responses to UK 14,304 were inhibited by rauwolscine (3x10‐8M) but not by prazosin (10‐7M). Therefore, phenylephrine, cirazoline or norepinephrine constrict mouse tail arteries by activating α1 and α2‐ARs. Based on antagonist potency, the order of selectivity for α1‐ARs is cirazoline > phenylephrine = norepinephrine.
The Effects of Acute Phase Proteins in the Blood‐brain Barrier and Tight Junction Proteins
Authors: Zhang Shuangling , Mark Karen S.
Published: June 2021

Tight junction (TJ) proteins, which form cell‐to‐cell contacts, play a central role in maintaining the integrity of the blood‐brain barrier (BBB) and brain homeostasis. Acute phase proteins (APP) are elevated in chronic inflammatory diseases but their effects in BBB function and structure have not been investigated.

The objective of this study was to examine the effects of APPs on an in vitro BBB model using rat brain microvessel endothelial cells (RBMEC). Following treatment with APPs, culture supernatant and protein were collected. The following methods were used in our study: enzyme‐linked immunoabsorbant assay (ELISA) to measure cytokines, Western blot analysis for TJ protein expression, and immunohistochemistry for TJ protein localization. Additionally, RBMEC permeability following APP treatment was measured.

Results from this study showed a significant increase of TNF‐α in the supernatant following treatment with α‐1‐acid glycoprotein (AGP). Increased permeability was seen with AGP treatment while serum amyloid A treatment induced decreased permeability. Our immunohistochemistry results showed a decrease in claudin‐5 expression following haptoglobin treatment.

In summary, our results show that APPs have direct effects on BBB function and TJ protein localization. This suggests that APPs may play a role in altering BBB integrity in chronic inflammatory diseases.

Funded by: NHLBI, R15‐HL092561
Cyclooxygenase Regulation Contributes to Hyperhomocysteinemia induced Endothelial Dysfunction in Transgenic Cystathionine beta‐synthase Deficient Mice
Authors: Cheng Zhongjian , Jiang Xiaohua , Durante William , Kruger Warren D , Yang Xiaofeng , Wang Hong
Published: June 2021

We examined the mechanism of hyperhomocysteinemia (HHcy) induced endothelial dysfunction in a novel severe HHcy mouse model, in which the mouse cystathionine β‐synthase (CBS) is deficient and an inducible human disease relevant CBS mutation (Tg‐S466L) is introduced to circumvent the neonatal lethality of the CBS deficiency (TghCBS‐S466L CBS−/−). Endothelium‐dependent vascular relaxation to acetylcholine (ACh) was impaired, and endothelium‐independent vascular relaxation to sodium nitroprusside was not changed in thoracic aorta from TghCBS‐S466L CBS−/− mice (plasma Hcy 180μm). HHcy‐impaired relaxation to ACh was restored by preincubating the aortic rings with a non‐specific cyclooxygenase (COX) inhibitor indomethacin (10 µM), a COX‐2 selective inhibitor nimesulide (1 µM), and a thromboxane A2/prostaglandin H2 (TXA2/PGH2) receptor antagonist SQ 29548 (10 µM), but not by a COX‐1 selective inhibitor SC 560 (10 µM). COX‐2 protein levels were increased in the aorta by severe HHcy in TghCBS‐S466L Cbs−/− mice. Finally, L‐Hcy (500 µM, 48 h) induced the levels of COX‐2 but reduced COX‐1 proteins in cultured primary human aortic endothelial cells. Our data suggest that HHcy impairs endothelial functions by inducing COX‐2 result in increased contractile TXA2/PGH2 production, and potentially, by reducing COX‐1 leading to decreased vasodilator prostacyclin production in the macrovasculature.
Angiotensin II elevates endothelial arginase activity via RhoA/MAPK pathways
Authors: Shatanawi Alia , Romero Maritza Josefina , Iddings Jennifer A. , Caldwell Ruth B. , Caldwell R. William
Published: June 2021

Elevated arginase (ARG) activity contributes to vascular endothelial dysfunction (ED) by decreasing L‐arginine availability to NOS and NO production and increasing reactive oxygen species. We previously showed elevated ARG activity/expression is involved in diabetic coronary ED in rats and in impaired NO production in bovine aortic endothelial cells (BAEC) exposed to high glucose. Angiotensin II (AngII) also contributes to progression of vascular complications in diabetes and other cardiovascular diseases. We have shown that exposing BAECs to AngII (0.1 μM, 24 hrs) elevates ARG activity/expression and decreases NO production; pretreatment with BEC, an ARG inhibitor, prevents these effects. These AngII effects are also prevented by simvastatin or RhoA kinase inhibitor Y‐27632 pretreatment, indicating ARG activation occurs via the RhoA pathway. Other studies indicate RhoA as a central upstream regulator of MAP kinase activity. We have found that AngII elevates active RhoA levels and phosphorylates p38 MAPK in a time and dose dependent manner. To further examine ARG activation pathways downstream of RhoA, we pretreated cells with p38 inhibitor SB202190 (5 μM). This treatment prevents AngII‐induced increased ARG activity/expression and maintains NO production. Together, these results strongly suggest AngII increases ARG activity/expression via activation of RhoA/MAPK pathways. Support: NHBL70215
Involvement of scavenging receptor‐B1 in NO release by insulin‐like growth factor binding protein‐3 (IGFBP3) in human endothelial and CD34 cells
Authors: Jarajapu Yagna , Kielczeski Jennifer , LiCalzi Sergio , Kent David , Grant Maria
Published: June 2021

We recently reported the vascular protective potential of IGFBP3 involving recruitment of circulating endothelial progenitor cells (CD34) to the sites of vascular injury. NO plays an important role in the migration of CD34 cells and therefore, we hypothesized that NO is involved in the effects of IGFBP3. NO release was quantified by DAF‐FM fluorescence in human cord blood‐derived CD34 cells, umbilical vein endothelial cells (HUVECs), pulmonary microvascular endothelial cells (HMVECs) and human retinal ganglionic cells (HRGCs). IGFBP3 (100 ng/ml) increased NO release in all four cell types. Pretreatment with scavenging receptor‐B1 (SRB1) antibody inhibited NO release in endothelial cells but not in HRGCs. HDL induced NO release in endothelial and CD34 cells but not in HRGCs in which mRNA for SRB1 was not detected. Co‐treatment with IGFBP3 and HDL caused a greater release of NO than with either individual treatment. DMS, a sphingosine kinase inhibitor blocked NO release in CD34 cells. IGFBP3 caused eNOS and vasodilator‐stimulated peptide (VASP) phosphorylation and redistribution of VASP in endothelial cells that was inhibited by L‐NAME. IGFBP3 and HDL caused relaxation of phenylephrine‐preconstricted rat mesenteric arterial segments that was inhibited by L‐NAME and combination of L‐NAME and indomethacin, respectively. These results suggest involvement of NO in the vascular protective effect of IGFBP3.
Dependence of Cathepsin L –induced Coronary Endothelial Dysfunction upon Activation of NAD(P)H Oxidase
Authors: Zhang Fan , Zhang Andrew Y , Li Pin‐Lan
Published: June 2021

Cathepsin L can generate endogenous endostatin in vascular and epithelial basement. The present study was designed to determine whether and how this cathepsin L‐derived endogenous endostatin alters endothelium‐dependent vasodilator responses in coronary arteries. In isolated and perfused small bovine coronary arteries, cathepsin L (200 ng/ml) markedly attenuated endothelium‐dependent vasodilator responses to bradykinin or A23187 by 56.16±9.58% and 68.95±10.32%, respectively. This inhibitory effect could be significantly reversed by pre‐treatment with O2−. scavenger, tiron, or anti‐endostatin antibody. ELISA assay revealed that cathepsin L dose dependently increased endostatin production in coronary arteries. In situ fluorescent microscopic imaging showed that cathepsin L decreased bradykinin‐ and A23187‐induced NO production in the intact endothelium, but with no effect on Ca2+ response. This cathepsin L‐induced reduction of NO was restored by the pretreatment of an anti‐endostatin antibody and tiron. Electron spin resonance (ESR) analysis demonstrated that cathepsin L increased O2−. production, which could be markedly attenuated by apocynin or anti‐endostatin antibody. It is concluded that endostatin could be endogenously produced in coronary arteries, which may result in endothelial dysfunction by NADPH oxidase activation (Supported by NIH grants HL57244, HL075316, and DK54927).