bluuebunny/crossref_metadata_2025 huggingface dataset:
Mechanism of AMPK suppression of LXR‐dependent Srebp‐1c transcription
Authors: Yang Jian
Published: June 2021

Activation of AMP‐activated protein kinase (AMPK) inhibits hepatic fatty acid synthesis by suppressing sterol regulatory element‐binding protein (SREBP)‐1c via inhibition of endogenous liver X receptor (LXR) ligand production and SREBP‐1c processing. However, whether AMPK directly inhibits ligand‐induced LXR activity remained undetermined. In this study we used a series of mutant Srebp‐1c promoter linked to a luciferase reporter to determine the inhibitory mechanism in rat hepatoma McA‐RH7777 cells. AMPK activation by either AICAR or metformin decreases Srebp‐1c promoter activity by about 75%. When endogenous LXR ligand production was blocked by the potent HMG CoA reductase inhibitor compactin, T0901317‐induced Srebp‐1c promoter activity was decreased by AICAR or metformin treatment. In the mutant Srebp‐1c promoter in which two LXR elements are intact but the sterol regulatory element (SRE) is disrupted, the fold inductions of the promoter activity by T0901317 without AMPK activators are significantly higher than those with AMPK activators. Furthermore, AMPK activation attenuates induction of endogenous SREBP‐1c mRNA by T0901317. These results indicate that AMPK directly inhibits ligand‐induced LXR activity in addition to blocking production of endogenous LXR ligands.
Enzymatic and structural studies of UDP‐2,3‐diacylglucosamine hydrolysis in lipid A biosynthesis
Authors: Young Hayley E. , Metzger Louis E. , Lee John K. , Stroud Robert M. , Zhou Pei , Raetz Christian R. H.
Published: June 2021

The outer‐leaflet of the outer membrane of Gram‐negative bacteria is composed of lipopolysaccharide (LPS), which is attached to the membrane via a hexa‐acylated saccharolipid called lipid A. The fourth step of the lipid A biosynthetic pathway, involving the conversion of UDP‐2,3‐diacylglucosamine (UDP‐DAGn) to 2,3‐diacylglucosamine 1‐phosphate (lipid X) and UMP, can be carried out by either LpxH or LpxI. These proteins do not share sequence homology, are never found in the same organism, and use different chemical mechanisms to catalyze the formation of lipid X. We have cloned the H. influenzae ortholog of LpxH (HiLpxH) in E. coli, purified it to 95% homogeneity, and determined its kinetic parameters and required cofactors in vitro. Mass spectrometry reveals that lipid X co‐purifies with HiLpxH. Interestingly, this property was also observed in the purification of C. crescentus LpxI, suggesting that the enzymes may share a product‐binding site. To further investigate this possibility, we have pursued structural studies of LpxH using a thermophilic ortholog from M. capsulatus (McLpxH). McLpxH has been overexpressed in E. coli and purified it to 95% homogeneity. Overall, this work sets the scene for further structural and mechanistic comparisons of LpxH and LpxI. This work is supported by NIH grant GM051310 to C.R.H.R. and Specialized Center grant of the Protein Structure Initiative U54 GM074929‐01 to R.M.S.
Cytosolic antioxidant enzyme regulate TNF‐alpha‐induced apoptosis
Authors: Lee Jooyoung , Kang Sangwon
Published: June 2021

Tumor necrosis factor α (TNFα) is a pleiotrophic cytokine which induces the activation of NF‐κB, MAP kinases, and cell death. TNF‐α‐induces the programmed cell death under the condition, where NF‐κB activation is defective, concomitant with robust increase of reactive oxygen species (ROS). Therefore, intracellular signaling processes including the inhibition of NF‐κB activation and robust increase of reactive oxygen species (ROS) are prerequisite for the induction of cell death programming. In order to elucidate the molecular mechanisms underlying the involvement of ROS in TNF α‐induced cell death, we targeted cytosolic antioxidant enzymes including catalase, glutathione peroxidase, and peroxiredoxin which are H2O2‐reducing peroxidases. Systematic depletion of cellular H2O2‐reducing peroxidases using RNA interference revealed that ablation of cytosolic antioxidant enzymes only augments both the biphasic H2O2 level and caspase‐dependent apoptosis induced by TNF‐α in HeLa cervical cancer cell line, but they did not modulate necrotic cell death. Although the depletion of any of the endogenous antioxidant enzymes results in global increase of the intracellular ROS level generated in response to TNF‐α‐induced apoptosis, PrxII regulatable H2O2 only influences caspase 8‐dependent apoptosis. Moreover, GPx I have a dominant effect on stress kinase activation and consequential cell death including p38 and JNK. Collectively, these results suggest that a novel mechanism underlying the selective action of intracellular ROS, precisely H2O2, in TNF α‐induced apoptosis. This study was supported by a grant of Discovery and validation of new drug project (2011‐0030195).
Strain differences in response to stress: Territorial urine markings, body weight, and temperature
Authors: Santos William Jose , Melige Monique , Chakraborty Nabarun , Meyerhoff James , Hammamieh Rasha , Jett Marti
Published: June 2021

We have developed a social‐stress (SS) mouse model using three different strains to better understand strain differences in response to stress. Retarded territorial urine marking (UM), obesity, and abnormal increments of body temperature are typical manifestations of stress in the rodent family. A comprehensive characterization of such stress responses across different mouse strains is still obscure and is addressed in the particular research scope presented here. In the present SS model, subject mice were individually housed in a small mesh box placed inside an aggressor's (AGG) cage for 6 hr/d x 10 d, food and liquid deprived, and exposed directly to the AGG 3x/d at random times. Controls (C) were simultaneously placed in a similar mesh boxes in the absence of AGG. Filter papers placed under the mesh boxes were collected at the end of the day and UMs were contoured via UV light exposure. SS mice of C57 showed significantly higher obesity and reduced UM vs. C that also gained weight; though a sign of habituation was apparent among C at the later part of 10 d. SS mice of BALB/c gained equivalent weight with C, but showed persistently reduced UM. In contrast, both C and SS of DBA lost weight over 10 d SS and showed increased UMs, suggesting a sign of hyper‐exploratory activity despite the stress. The current findings indicate that distinct strains of mice react differently to SS.
Proteasome Inhibitor Epoxomicin Prevents Cardiomyocyte Apoptosis through Regulation of Gene Expression
Authors: Li Dan , Babu Gopal
Published: June 2021

Epoxomicin (E) is shown to prevent apoptosis during myocardial stress. In this study, we aimed to identify the molecular mechanism(s) associate with the anti‐apoptotic effect of E, using neonatal rat ventricular myocytes as a model system. The cultured cardiomyocytes were pretreated for 24 hrs with E. Apoptosis was induced by chelerythrine (Che) (5μM) for 35 min. Interestingly, Che induced apoptosis is associated with significant Akt phosphorylation at ser473. We found that E dose‐dependently (0.1μM) protects cardiomyocytes against apoptosis. Pretreatment of cardiomyocytes with E prevents Akt Ser473 phosphorylation and, thereby, apoptosis. The decrease in Akt Ser473 phosphorylation by E correlates with its protective effect in a time dependent manner. The phosphorylation of Akt at Thr308 was not altered by Che and/or by E treatment. To determine if the Akt Ser473 phosphorylation is proapoptotic, we ablated the expression of Rictor, a key component of mTORC2, which phosphorylates Akt at ser473. Rictor knockdown by RNA interference dramatically inhibits the Akt ser473 phosphorylation in Che treated cells, and reproduces the protective effects of E. Gene expression analysis shows that the expression of Akt substrate1 (Akt1S1), a negative regulator of mTORc kinase was significantly up‐regulated in E treated cells. These results together suggest that E inhibits the phosphorylation of Akt ser473 through negative regulation of Rictor activity. We further investigated the expression and phosphorylation patterns of upstream and downstream regulators of the Akt pathway. Our results show that Che activates the MEK/ERK signaling pathway and this activation was inhibited by E or by Rictor knockdown. In conclusions, our studies demonstrate that (1) the Akt activation, which is known to promote cell growth and survival, can also trigger cell death under certain conditions, and (2) in addition to the proteasome inhibition, E protects cardiomyocytes against apoptosis by changing gene expression.
Tuberculosis and Epigenetics: Functional Noncoding RNA in the Host‐Pathogen Interaction
Authors: Wissler Josef H
Published: June 2021

OBJECTIVE

Noncoding ncRNAs, the main RNA transcript fraction, were investigated from tuberculin‐activated mononuclear cells [MMC] for functions in host‐pathogen interactions and phenotype changes upon transfer of the tuberculin reaction as a feature of transmissible adaptive cell‐mediated immunity.

METHODS

Ann. N. Y. Acad. Sci. 1022 : 163–184, 2004; 1137 : 316–342, 2008; OWWZ 2010: http://www.owwz.de/868.html?&L=1

RESULTS

The MMC activation by purified protein derivative tuberculin [PPD] and assays in use represent in vitro correlatives of delayed‐type [−IV] hypersensitivity reactions including Koch‐Metchnikoff‐ and Jones‐Mote types. These concern induration, swelling and endothelial infiltration of memory MMC into skin lesion sites within 24 to 72 hours. PPD‐activated cells form ncRNA. Some modified, edited, redox‐ and metalloregulated functional small hairpin fncRNA [<200n] were isolated and sequenced. Modified nucleosides are e.g. isoguanosine / crotonoside, adenosine‐N1‐oxide of 151Da base families. fncRNAs regulate phenotypic adaptation by complementary interaction with defined conserved homologous helix‐nucleating Mendelian consensus domains shared in epigenetic regulator proteins entangled in tolerated growth, vascularization, metabolic syndromes, cancer, epi‐ and genetic information indexing of the epigenome.

CONCLUSIONS

fncRNA from PPD‐activated cells are considered useful for better evaluations of test systems of infectious diseases, nuclear antibodies, aberrant histone codes in pathogenesis of autoimmune disease [e.g. SLE] and tolerance of biomaterials. Their features suggest that immune cells beside known protein recognition systems, may be armed by additional arsenals of recognition entities in terms of found fncRNAs
An assessment of the actual nutrition practice in elderly
Authors: Abduldaeva Aigul , Dalenov Erbolat
Published: June 2021

The study progression of the elderly population in the world and an increasing mortality rate of old age diseases ‐ cardiovascular system, diabetes, neurodegenerative processes, reducing resistance to infections, malignancies contributes to the relevance of healthy nutrition issues for human body. The purpose of the study is an assessment the actual nutrition practice among elderly living in Astana. The study involved 54 people aged 60 – 70 years old living in Astana. The method of 24‐hour diet recall and statistics methods were used. The analysis of dietary intake is revealed that the average caloric content of older people diet in Astana was about 2000 kcal, primarily due to animal protein. Whereas, the content of dietary fiber in the diet of majority older people is only 22% of the recommended standards, and only a minority of elderly dietary fiber content of 55% of the recommended norms. The content of vitamins A, group B, C, PP, E ranges from 37% to 53% of the recommended norms, which causes high morbidity rate in this population. This implies that there should be promotion of healthy nutrition and proper eating behavior model of the population since early age and throughout life. Preservation of high caloric intake in old age affects human health. Thus, studies have shown a direct relationship between the health of older people and their diet.
Notch1 ablation induces molecular changes in plasticity genes that contribute to memory formation
Authors: Alberi Lavinia , Gaiano Nicholas
Published: June 2021

Beside the essential role in development Notch signaling has been involved in neuronal morphology, synaptic plasticity, learning and memory. In our previous work we reported that Notch is induced in hippocampal networks following spatial exploration. In addition targeted loss of function of Notch in the mature hippocampus has confirmed that this pathway is required for synaptic plasticity and novel memory acquisition. The mechanism beyond Notch function remains to be fully elucidated. Here we present several lines of changes in plasticity‐associated genes following ablation of Notch1. Notch1cKO hippocampi show a strong decrease in NMDARs as well as GluRs. As a consequence of reduced NMDAR's levels, we also observe a decrease in CamKII expression, which is know to modulate NMDAR dependent plasticity. At the level of secondary messengers, that mediate the molecular changes associated with memory formation, we show that phosphorylation of ERK is reduced, and that the total levels of CREB are also decreased. In Notch1cKO aging mice we observe a profound loss of dendritic spines without any major effect on the dendrites’ length, on the other hand the dendrites are characterized by abundant varicosities, indicative of neurodegeneration. These finding suggest that, in the Notch1cKO mice, the molecular players of memory are profoundly altered, and that in older mice signs of degeneration are present.
Factors Associated with Osteoporosis in Korean Postmenopausal Women: Data from the Korean National Health and Nutrition Examination Survey (KNHANES) 2009
Authors: Kim Mi-Hyun , Johnson Mary Ann , Lee Jung Sun
Published: June 2021

As the population ages, osteoporosis is a growing global public health problem. It is of great importance to identify modifiable risk factors of osteoporosis to promote bone health of the older population. This study examined the risk factors associated with osteoporosis in a nationally representative sample of Korean postmenopausal women from 2009 KNHANES (n=1,480, means age±SE=61.2±0.3). Bone mineral density of total femur, femoral neck, and spine was measured by X‐ray absorptiometry. Osteoporosis was determined as T‐score of −2.5 or below in at least 1 of 3 sites. Menopausal status was confirmed by self‐reports. Around 43% of the study sample had osteoporosis, and poor socioeconomic status, lower BMI, and shorter estrogen exposure duration were significantly associated with osteoporosis. Poor dietary intake was also related to osteoporosis. In the age‐adjusted logistic regression models, participants consuming protein, vitamin B2, or vitamin C less than EAR showed higher odds of having osteoporosis than their counterparts. Participants consuming no fish/shellfishes or no milk/milk products had 32% and 46% increased odds of having osteoporosis, respectively. The findings from this study suggest several modifiable risk factors that can be addressed in the development and implementation of tailored nutritional intervention programs to improve bone health of Korean postmenopausal women.
Measured vs. Estimated Resting Energy Expenditure in a Clinic Population of Obese Youth
Authors: Henes Sarah , Hickner Robert , Cummings Doyle , Houmard Joseph , Kolasa Kathryn , Lazorick Suzanne , Collier David
Published: June 2021

The objective of this study was to compare measured resting energy expenditure (MREE) using portable indirect calorimetry in obese youth with that estimated from six published predictive equations used by clinicians to determine caloric targets for weight loss.

MREE was measured in fasting obese youth who were referred to a multidisciplinary Healthy Weight Clinic (n=80, mean age 12.18 yrs ± 0.31 SE, mean BMI 37.39 ± 1.07 SE, mean BMI z score 2.47± 0.04 SE). The mean caloric difference between MREE and each predictive equation was calculated. The Harris Benedict equation demonstrated the lowest mean caloric difference (197.94 kcal/d ± 23.26 SE) compared to MREE. While there was a strong correlation (p < 0.05) between MREE and estimations from each of the predictive equations, there was substantial variability between estimations and discrepancies by gender. Mean measured and estimated caloric differences were greater for males (634.12 kcal/d± 69.45 SE) than for females (254.51 kcal/d± 25.92 SE)(p<0.05). No significant racial differences were evident. These data demonstrate the inconsistent findings among predictive equations and suggest a stronger role for portable indirect calorimetry with obese youth in the clinical setting. When indirect calorimetry is not available, the Harris Benedict equation may be the best predictive equation to use in obese youth aged 7–18 who are >99th percentile for age and gender.
Perceptions of Key Parental Benefits from Practices that Promote Intake of Calcium‐Rich Foods and Beverages (CRF/B) in Preadolescent Children
Authors: Wong Siew Sun , Gunther Carolyn , Reicks Marla , Richards Rickelle , Bruhn Christine M , Cluskey Mary , Misner Scottie , Watters Corilee , Bellajos Miriam
Published: June 2021

Objective

Explore perceived benefits underlying parent practices that promote CRF/B intake by preadolescents (10–13y), including availability, setting rules and expectations, and role modeling.

Methods

Focus groups using Nominal Group Technique with multi‐ethnic parents of preadolescents and qualitative data analysis methods.

Results

In seven states (AZ, CA, HI, MN, OH, OR, UT), 156 parents (83% mothers; 22% African American, 20% Asian, 26% Hispanic, 27% non‐Hispanic White) completed the study. The top 3 perceived benefits were similar across parenting practices and included:

Making CRF/B Available
Rules & Expectations
Role Modeling

Child's bone/teeth health & growth
Child's general health
Child's health

Family's healthy eating habits/choices
Parent's positive emotions
Family's general health

Child's mental health & school performance
Child's current bone health
Family's dietary intake/choices

Conclusion

Having a healthy child was the primary perceived parental benefit of practices that promoted calcium intake by children. Secondary benefits were a healthy family diet and parent's positive emotions.

Source of Research Support: USDA NIFA, AES & Extension at participating universities.
Developing and pre‐testing nutrition messages for Asian Indian mothers
Authors: Momin Shabnam R , Olson Beth
Published: June 2021

Asian Americans born in the US are twice as likely to be overweight compared to those born elsewhere and newly migrating. The prevalence of childhood overweight for Asian Indians will increase as more Asian Indian children are born in the US. Childhood obesity prevention efforts often focus on parent behaviors that influence children's dietary intake and weight status. Research shows that Asian Indian mothers may need nutrition education materials developed specifically for this group. The aim of this study was to develop and pre‐test nutrition messages for Asian Indian mothers addressing eight parental food‐related behaviors likely to influence children's diet and weight status. Nutrition education messages developed by USDA were used with some adaptations and new messages were developed as needed based on the qualitative work previously completed with Asian Indian mothers. These messages were then tested to assess their effectiveness. Mothers were receptive to most messages. Minor revisions were made to some messages to improve clarity and increase acceptability. Messages that received strong negative responses were deemed inappropriate for this population. Nutrition professionals working with Asian Indians could use the final messages to inform mothers about developing healthy eating habits in their children.

Funded by: MSU AgBioResearch and USDA Multistate Project W1005

Grant Funding Source: MSU AgBioResearc and USDA Multistate Project W1005
Stepping Up to the Challenge: the development, implementation, and assessment of a statewide, regional leadership program for child nutrition directors
Authors: Bergman Jacqueline Joanne , Beall Deborah , Zidenberg-Cherr Sheri
Published: June 2021

A statewide, regional leadership training program was developed, implemented and assessed throughout California for child nutrition directors with the goal of creating a healthy school environment (HSE) and regional networks for collaboration and HSE sustainability. Needs of the target audience were identified through literature review, expert interviews and responses to a needs assessment (NA) questionnaire. Qualitative analysis of the NA (n=256) identified (1) planning cost‐effective menus, (2) reducing calories, sodium, saturated fat, trans‐fat in menus and (3) using United States Department of Agriculture Foods cost‐effectively to enhance menus as the top three training topics. Top training topics were incorporated into a two‐day training by using social ecological and marketing methods in a train‐the‐trainer format. Evaluation of a follow‐up questionnaire identified top “insights, inspirations and strategies” as (1) marketing of school foods program, (2) expansion of salad bars and (3) collaboration with community partners for an improved school environment. Analysis also indicated a need for further training assistance on procuring produce from local farmers, menu planning, meeting Healthier US School Challenge criteria and preparing for proposed meal pattern rules. This information will be used to develop technical assistance sessions. Funded by USDA 2010 Team Nutrition Training Grant.
Fat distribution, insulin resistance and serum adiponectin of older women with and without metabolic syndrome
Authors: Chongwatpol Pitipa , Hermann Janice R , Bogale Alemtsehay , Stoecker Barbara J
Published: June 2021

This study assessed associations among dietary intakes, body fat distribution and serum biomarkers in 41 women aged > 50 yrs with and without metabolic syndrome (MetS). MetS by International Diabetes Federation definition (IDF) was waist circumference ≥88cm with two additional factors: elevated blood pressure, triglycerides, or glucose; or decreased HDL‐cholesterol. Three‐day diet records and 24‐hr duplicate plates were collected. Dual‐energy X‐ray absorptiometry (DEXA) scans was used to estimate body fat distribution. Total antioxidant capacity (TAC) in diet and serum were measured with the ferric reducing antioxidant power (FRAP) assay. The MetS group had significantly higher serum insulin and adiponectin as well as higher homoeostasis model assessment (HOMA)‐β, HOMA‐IR, and android:gynoid ratio. TAC in serum and 24‐hr duplicate plate diet samples were not significantly different between control and MetS groups; however, dietary fiber intakes were positively associated with dietary vitamin C (p<0.03). Total body fat, android fat, android:gynoid ratio, and serum adiponectin were significantly (p<0.05) associated with insulin resistance in these older women. (Supported by Oklahoma Agricultural Experiment Station, W2002).
Broccoli‐derived compounds modulate androgen up‐regulation of C‐C chemokine ligand 2 and enhancement of monocyte attraction to prostate cancer cells
Authors: Kim Eun-Kyung , Kim Young S. , Yu Liangli , Milner John A. , Wang Thomas T.Y.
Published: June 2021

Inflammation plays a key role in prostate tumorigenesis. Recruitment of inflammatory monocytes to the tumor site is mediated by C‐C chemokine ligand 2 (CCL2) through binding to its receptor CCR2. We hypothesized that androgen could modulate CCL2 expression in hormone‐responsive prostate cancer cells and thereby promote recruitment of monocytes. Given the inhibitory effect of broccoli‐derived compounds indole‐3‐carbinol (I3C) and 3,3′‐diindolylmethane (DIM) on androgen‐dependent pathway, we also reasoned that I3C and DIM could modulate the effect of androgen on CCL2‐mediated pathways. We found that androgen dihydrotestosterone (DHT) induced time (0–72 hrs) and concentration‐dependent (0–1 nM) increases in CCL2 mRNA levels in the androgen‐responsive human prostate cancer cell LNCaP. The increase in CCL2 mRNA corresponded to increased secretion of CCL2 protein by LNCaP cells. This effect of DHT was mediated through an androgen receptor (AR)‐dependent pathway as small inhibitor RNA against AR negated the induction of CCL2 mRNA by DHT. Moreover, conditioned media from androgen‐treated cells promoted human monocyte THP‐1 cell migration. Both I3C and DIM inhibited promotional effects of DHT on CCL2 and migration. These results suggested that androgen regulates CCL2 and promotes inflammatory micro‐environments in prostate tumors, and this process can be regulated by broccoli‐derived compounds.
Anti‐angiogenic Effects of Citrus Polymethoxyflavones and Their Major Metabolites
Authors: Nutakul Wasamon , Zheng Jinkai , Shao Rong , Xiao Hang
Published: June 2021

Previous studies have demonstrated inhibitory effects of citrus polymethoxyflavones, nobiletin (NBT) and 5‐hydroxy nobiletin (5HN) on several types of cancer. We have identified major metabolites of NBT (i.e. M1: 3′‐hydroxy NBT; M2: 4‐hydroxy NBT; and M3: 3′,4′‐dihydroxy NBT) and 5HN (i.e. M4: 3′,5‐dihydroxy NBT; M5: 4′,5‐dihydroxy NBT; and M6: 3′,4′,5‐ trihydroxy NBT) in rodents fed with NBT and 5HN. In this study, we investigated the anti‐angiogenic activities of NBT, 5HN and their major metabolites using wound healing and tube formation assay with Human microvascular endothelial cells. Among NBT metabolites, M2 showed strongest inhibitory effects, while among 5HN metabolites, M4 had the most potent effects. In wound healing assay, M2, M1, NBT and M3 at 50 μM inhibits 50%, 40%, 30% and 10% of cell migration, respectively. M6, M5 and 5HN at 10 μM inhibited 50%, 40% and 30% of cell migration, respectively. Moreover, M4 at only 3μM inhibited more than 90% of cell migration. In tube formation assay, M3 and NBT at 50 μM inhibited 75% and 30% of capillary tubes formed, respectively, while M1 did not inhibit tube formation. M6, M5 and 5HN at 10 μM inhibited 90%, 77%, 54% of tube formation, respectively. Most interestingly, M2 at 50 μM and M4 at 3μM abolished all tube formation. Our results suggested that the number and position of hydroxyl groups in NBT, 5HN and their metabolites dictate their inhibitory effects on angiogenesis.

Grant Funding Source : NIH: CA139174, AICR: 10A044
Fibrinolytic Resistance and Prothrombotic Phenotype is Altered in Mice Lacking α(1,3)‐Fucosyltransferase‐IV and – VII
Authors: Wang Huili , Mackey Lantz , Rose Jason , Homeister Jonathon W
Published: June 2021

Mice deficient in α(1,3)‐fucosyltransferase type‐IV and ‐VII (DKO) lack selectin‐type leukocyte adhesion molecule ligand activity, and show a prothrombotic phenotype in a model of carotid artery injury that is associated with enhanced platelet aggregation. We sought to confirm the prothrombotic phenotype in a second experimental model, and examine whether it was in part due to increased procoagulant activity. The phenotype was examined using an in vivo model of collagen‐epinephrine‐induced pulmonary thromboembolism. Plasma procoagulant activity was assessed by calibrated automated thrombography (CAT), fibrinogen ELISA, and a tissue‐type plasminogen activator (tPA)‐ induced thrombolysis assay. DKO mice had more pulmonary thromboemboli and a higher mortality rate than wild‐type (WT) controls or mice deficient in P‐selectin glycoprotein ligand‐1. Thrombin generation in DKO mice was not different from WT mice, but DKO mice had a 30% higher plasma fibrinogen concentration. tPA‐dependent fibrinolysis, measured by release of labeled fibrinogen degradation products, was reduced in DKO whole blood clot compared to wild‐type clot. WT clot dry weight after lysis was reduced 38%, but DKO clot weight was reduced only 9%. These data confirm the selectin‐independent prothrombotic phenotype of DKO mice and suggest that it is due in part to increased plasma fibrinogen and decreased fibrinolysis.
Cavin1 deficiency results in abnormal vascular function in mice
Authors: Rippe Catarina , Albinsson Sebastian , Swärd Karl
Published: June 2021

Caveolae are membrane microdomains which play a role in vascular function. Cavin‐1/PTRF is a newly discovered protein which, in concert with the caveolins, is involved in caveola biogenesis. Our aim was to examine vascular function in mice lacking cavin‐1. We used pressure myography to examine myogenic tone and charbachol‐induced dilation in the presence and absence of L‐NAME. Blood pressure was measured in the carotid artery of anaesthetized mice. Cavin‐1‐deficient mice had significantly lower myogenic tone compared to WT (5±3% vs. 22±6% P<0.01). In the presence of L‐NAME, myogenic tone was restored towards control levels. Vessel diameter at 45mmHg was significantly larger in the cavin‐1 KO mouse compared to control (329±7 vs. 259±9 μm, P<0.05). Blood pressure was slightly lower in cavin‐1‐deficient mice (87± 7 vs. 114±9 mmHg, P<0.05). We show for the first time that cavin‐1 deficient mice exhibit abnormal vascular function, due in part to excessive NO production, and with direct consequences for the systemic blood pressure level. This work was supported by the Swedish Research Council and the Crafoord Foundation.
Non‐antithrombin affinity Semuloparin and it's component oligosaccharides release endogenous TFPI in a molecular weight dependent fashion
Authors: Fareed Jawed , Khan Hussein , Litinas Evangelos , Cunanan Josephine , Hoppensteadt Debra , Viskov Christian
Published: June 2021

Semuloparin (S) is an ultra low molecular weight heparin (uLMWH) with a unique composition. The purpose of this study was to determine the effect of non‐AT affinity semuloparin (NATAS) and it's oligosaccharide components on TFPI release. Non‐human primates (n=4–6; Macaca mulatta) were administered with 1.0 mg/kg IV, S, it's non‐AT affinity fraction and isolated hexasaccharide (H), dodecasaccharide (D) and octasaccharide (O) components. Citrated plasma samples were drawn at baseline and 15 minutes post‐administration. TFPI antigen levels by ELISA and tissue factor mediated thrombin generation using a fluorogenic assay were measured. S produced a sizable release of TFPI (156 ± 15 ng/ml). NATAS produced a slightly lower but statistically significant release of TFPI (142 ± 12 ng/ml). The oligosaccharide components produced a molecular weight dependent release of TFPI, with H producing minimal TFPI release (86 ± 8 ng/ml) vs. saline (78 ± 9 ng/ml). D produced a statistically significant increase in the release of TFPI (116+15 ng/ml). While S exhibited a measurable anti‐Xa response and produced a strong inhibition of thrombin generation (84% inhibition) whereas NATAS and it's fractions produced < 20% inhibition of thrombin generation. These results suggest that the NATAS and it's oligosaccharide components enhance TFPI antigen release which may mediate some of it's pharmacodynamic actions.
The Effects of Site of Surgical Implantation on the Delivery Profiles of Anti‐Trypanosomal Agent (DFMO)
Authors: Benghuzzi Hamed A , Tucci Michelle , Cameron Joseph A
Published: June 2021

This investigation was designed to compare the interface zone of the tricalcium phosphate lysine delivery devices loaded with diflouromethyl ornithine (TCPL‐DFMO) that were implanted either I/P or S/C. A total of 120 adult male rats were randomly divided into four groups. Each rat in groups 1–2 were implanted with TCPL‐DFMO devices I/P and S/C respectively and group 3 animals were implanted with empty TCPL (sham). Group 4 animals served as intact controls. At the end of 1, 4 and 10 weeks postimplantation10 animals from each were sacrificed following standard lab protocols and capsules were retrieved and blood analysis were performed. The results indicated that regardless of the site of surgical implantation there was a fibrous capsule which encompasses the implanted delivery system, and the presence of inflammatory cells were evident after the first week of implantation. The capsules implanted I/P experienced a greater portion of tissue growth and a higher release profile of DFMO (5–8 ng/ml/day) than those implanted S/C (2.7–4.8 ng/ml/day). In addition, the weights of the capsule and the weight of the tissue retrieved from S/C implants were significantly less than those implanted I/P. This information is necessary for future designs of the delivery devices, so that the prolonged delivery of DFMO from the TCPL is maximized.
Mechanisms involved in the translocation of AGS3 from cell cortex to the Golgi apparatus following activation of a G‐protein coupled receptor
Authors: Oner Sukru Sadik , Lanier Stephen M
Published: June 2021

AGS3 and related GPR proteins provide unexpected regulatory mechanisms for G‐protein signaling systems, which are involved in diverse various cellular functions. The AGS3·Gαi signaling module is apparently regulated by cell surface, seven‐transmembrane receptors as well as non‐receptor guanine nucleotide exchange factors. Upon activation of a cell surface receptor, AGS3 dissociates from the cell cortex and apparently moves to the Golgi apparatus (GA). The mechanism of this translocation and the functional consequences are not defined and this report focuses on these questions. The receptor‐mediated movement of AGS3 to the GA required the TPR domain of AGS3. AGS3 constructs lacking S/T phosphorylation sites in the GPR domain or containing site‐directed mutations in the TPR domain known to alter AGS3 trafficking were effectively relocated to the GA following receptor activation. Co‐expression of the nonreceptor GEF Ric‐8A also disrupted the AGS3·Gαi signaling module, but did not increase the presence of AGS3 in the GA. Finally, receptor‐mediated movement of AGS3 to the GA altered the structural integrity of the GA as determined by immunofluorescence imaging of GA resident proteins. The regulated oscillation of AGS3 between the cell cortex and the Golgi apparatus offers unexpected mechanisms for modulating protein secretion and Golgi dynamics.
Evidence for a limbic cortical HPA‐inhibitory network and its role in chronic stress‐induced HPA axis hyperactivity
Authors: Radley Jason
Published: June 2021

A network of interconnected cell groups in the limbic forebrain regulates hypothalamic‐pituitary‐adrenal (HPA) activation during emotionally stressful experiences, and disruption of these systems is broadly implicated in stress‐related psychiatric and systemic illnesses. A significant challenge to progress has involved unraveling the circuitry and mechanisms providing for regulation of HPA output, as these limbic forebrain regions do provide any direct innervation of HPA effector cell groups in the paraventricular hypothalamus. While the current view involves a layer of cell groups providing an array multisynaptic parallel pathways between the limbic forebrain and PVH, recent advances in our research will highlight evidence for a discrete region within the bed nucleus of the stria terminalis that serves as a neural hub for receiving and integrating these influences. Insights gleaned from clarifying the mechanisms of HPA control during acute emotional stress have provided a framework for ongoing studies examining the relationship between chronic stress‐induced plasticity in key cell groups (notably, the medial prefrontal cortex) and HPA axis hyperactivity.
Etanercept, a tumor‐necrosis factor (TNF‐α) inhibitor, improves endothelial function of contralateral middle cerebral artery after cerebral ischemia in hypertensive rats
Authors: Girgla Saavia Singh , Pires Paulo Wagner , McClain Jonathon Lee , Dorrance Anne McLaren
Published: June 2021

Middle cerebral artery (MCA) occlusion (MCAO) causes endothelial dysfunction in the ischemic MCA (Marrelli et al., AJP 276:H33‐H41), and in small mesenteric arteries (Martinez‐Revelles et al., JPET 325: 363–9). The effect of MCAO on the contralateral MCA (CL MCA) however is unknown. In hypertensive rats MCA remodeling impairs artery function, and etanercept (ETN) attenuates this. We hypothesized that CL MCA dilation is impaired after MCAO and that ETN will improve this. 12 week old stroke prone spontaneously hypertensive rats, ETN‐treated from 6 weeks of age (ETN IR) or untreated (IR), underwent 1 hour of MCAO and 47 hours reperfusion. SHAM controls were generated. Cerebral blood flow (CBF) was measured by laser Doppler flowmetry. CL MCAs were mounted in a pressure myograph under physiological conditions. Endothelial function was assessed by intraluminal perfusion of ADP. Data are presented as mean ± SEM. Tone generation was not different between the groups. ADP mediated dilation was impaired in the IR group and this was ameliorated by ETN treatment (p<0.05). This caused a small, but insignificant (p=0.12), increase in post‐MCAO CBF in the ETN IR group. Our data suggest that ETN improves CL MCA endothelial function and may improve post‐MCAO CBF in hypertensive rats. (AHA0840122N)

SHAM (n=2)
ETN IR (n=3)
IR (n=4)

Tone (%)
32.6±5.4
33.2±7.1
36.9±4.7

Maximum increase in lumen diameter with ADP (μm)
62.5±12.5
60.6±7.5
28.2±7.8

Flow (% of pre‐MCAO)
94.5±10.9
86.6±7.0
62.6±10.1
Apocynin treatment attenuated middle cerebral artery remodeling in life‐long obesity in Sprague‐Dawley rats
Authors: Pires Paulo Wagner , McClain Jonathon Lee , Dorrance Anne McLaren
Published: June 2021

Metabolic syndrome is a disorder that includes abdominal obesity and hypertension, and these increase stroke risk. Diet‐induced obesity and hypertension lead to remodeling of the middle cerebral artery (MCA) through a mechanism dependent on superoxide anion (O2•−). In the vasculature, most O2•− is generated by the enzyme NADPH‐oxidase (Nox). We hypothesized that Nox inhibition with apocynin (Apo) would attenuate obesity‐induced MCA remodeling in rats. Three‐week old male Sprague‐Dawley rats fed a high‐fat diet (HF) for 17 weeks were treated with Apo (HF+Apo, n=7) or vehicle (HF, n=12). Rats fed normal chow were controls (NC, n=11). MCAs were mounted on a pressure myograph to assess passive structure. Data are mean ± SEM, NC vs HF vs HF+Apo. Rats fed HF had higher visceral fat:body weight (1.42±0.13, 3.92±0.19, 4.27±0.45, p<0.05). Apo prevented the HF‐induced increase in mean arterial pressure (126±3, 146±6, 125±3mmHg, p<0.01). At 80mmHg intralumenal pressure, Apo increased MCA lumen diameter (266±6.6, 245±8.3, 280±8.3μm, p<0.05), decreased wall thickness (20.2±1.5, 33.0±2.8, 18.0±1.2μm, p<0.05) and wall‐to‐lumen ratio (0.08±0.01, 0.14±0.01, 0.07±0.01, p<0.05). HF±Apo had no effect on distensibility (56.9±5.6, 48.8±4.6, 50.3±4.3). These data suggest that Apo prevented the increase in blood pressure in rats fed HF. This might be responsible for the beneficial effects of Apo in the MCA structure.
Regulation of K+‐induced [3H]D‐Aspartate Release by Synthetic Neuroprostanes in Isolated Bovine Retina
Authors: Flowers Arnecia Faye , Floyd Namonique , Kegey Edem , Jamil Jamal , Durand Thierry , Galano Jean-Marie , Guy Alexandre , Fatou Njie-Mbye Ya , Kulkarni Madhura , Ohia Sunny E , Opere Catherine A
Published: June 2021

Neuroprostanes (nPs) are a series of isoprostane‐like compounds that are spontaneously derived by free‐radical catalyzed peroxidation of the polyunsaturated fatty acid, docosahexanoic acid. Although nPs are elevated in neurodegenerative conditions (Musiek et al., Brain Pathol. 15:149,2005), their biological effects remain largely unknown. We investigated the role of synthetic nPs, CO5‐667, CO5‐668 and CO5‐738 on K+‐induced glutamate release (using [3H]D‐aspartate as a marker) in isolated bovine retina. Isolated neural retina were incubated in oxygenated Krebs solution containing 200nM of [3H] D‐aspartame for 60 mins and then prepared for studies of neurotransmitter release using the superfusion method. Release of [3H]D‐aspartate was evoked by iso‐osmotic concentration of K+ (50mM)‐stimuli applied at 80–88 mins (S1) and 116–124 mins (S2) after the onset of superfusion. In the concentration range, 1 nM to 10 μM, the nPs enhanced K+‐induced [3H]D‐aspartate release from retina without affecting basal tritium overflow. For instance, C05‐668 achieved a maximal excitatory response of about 80% (p<0.01, n=4) at the 10 nM concentration of the nP. At an equimolar concentration of 10 nM, the rank order of activity was as follows: CO5‐668> CO5‐738> CO5‐667. In conclusion, the novel synthetic nPs exert an excitatory effect on K+‐evoked [3H]D‐aspartate release in isolated bovine retina.
Effects of captopril on factors affecting gastric mucosal integrity in aspirin‐induced gastric lesions in Sprague‐Dawley rats
Authors: Ismai Nafeeza Mohd , Aziz Ibrahim Abdel , Jaarin Kamsiah
Published: June 2021

Captopril, an angiotensin converting enzyme inhibitor, used as an antihypertensive agent has been shown to have antioxidant properties which prompted the present study. This study investigated the effects of captopril on factors affecting gastric mucosal integrity in rats that had aspirin‐induced gastric lesions. Eighteen male Sprague‐Dawley (200–250g) rats given aspirin (40 mg/100g bw) were divided into a control group, captopril (1 mg/100g bw daily) and ranitidine (2.5 mg/100g bw bidaily) groups. Oral ranitidine and captopril were given for 28 days. Compared with the control group, captopril reduced gastric acidity (P<0.05), increased gastric GSH (P<0.05) and prostaglandin E2 (P<0.05). Ranitidine healed the lesions significantly (P<0.05) compared to the control group. There was, however no difference between ranitidine and captopril on the severity of lesions, gastric acidity, MDA and GSH. Captopril increased PGE2 compared to ranitidine (P<0.05). In conclusion, we found that captopril produced desirable effects on gastric acidity, PGE2 and GSH. Its capability to enhanced gastroprotective protective parameters and reduce the aggressive acid‐factor optimises the balance between protective and aggressive factors. In view of this linkage, we strongly suggest captopril as a potential novel antiulcer agent. Research was funded by the Faculty of Medicine, Universiti Kebangsaan Malaysia.
Sirtuin‐1 gain of function increases endothelium‐dependent vasorelaxation of murine femoral arteries
Authors: Calvert Robert , Noh Junghyun , Pereira Renata , Ruan Ting , Doo-Hyun Kwon , Zhang Quan-Jiang , Accili Domenico , Abel E Dale , Symons J David
Published: June 2021

Sirtuin‐1 (SirT1) is a NAD+‐dependent class III histone deacetylase that influences the functional properties of target proteins e.g., endothelial nitric oxide (NO) synthase (eNOS). Physiological (e.g., aging) and pathophysiological (e.g., diet‐induced obesity) processes can reduce arterial SirT1 expression, increase acetylated eNOS, decrease phosphorylated eNOS (p‐eNOS) to total eNOS, and impair endothelium‐dependent vasorelaxation. We tested the hypotheses that SirT1 overexpression increases arterial p‐eNOS to total eNOS and improves arterial function. Myocardial tissue from 8‐week old male SirT1 bacterial artificial chromosome overexpressor mice (TG) displayed 3‐fold increases (p<0.05) in SirT1 activity vs. wild‐type (WT) littermates (n=7 mice per group). Arterial reactivity was assessed in two femoral artery segments (~190 um, internal diameter) from each mouse using isometric tension techniques, and the entire aorta, iliac, and remaining femoral tissue was homogenized for immunoblotting. In arteries precontracted to 65% of Lmax tension, acetylcholine (ACh)‐evoked vasorelaxation was greater (p<0.05) in vessels from TG vs. WT mice, while responses to sodium nitroprusside were similar between groups. Improvements in ACh‐evoked vasorelaxation in TG vs. WT mice were negated (p<0.05) by NOS inhibition using L‐NMMA. p‐eNOS( S)1177 to total eNOS was greater (p<0.05) in vessels from TG vs. WT mice, while p‐Akt(S)473 to total Akt and p‐ERK1/2 to total ERK were similar between groups. These data indicate that mice with 3‐fold overexpression of SirT1 exhibit improved endothelial function and could be afforded vascular protection in response to aging and/or obesity.
Cytomegalovirus promotes arteriolar dysfunction through impairment of nitic oxide‐ and prostacyclin‐dependent vasodilation
Authors: Brunson Jerry , Stokes Karen Y.
Published: June 2021

Cytomegalovirus (CMV) has been implicated in cardiovascular disease, possibly by causing endothelial dysfunction. We have shown that persistent infection results in impaired endothelium‐dependent arteriolar dilation, in part due to a COX‐1‐dependent reduction of COX‐2‐mediated dilation. Our objective was to further clarify how CMV leads to arteriolar dysfunction, by assessing the roles of prostacyclin (PGI2), thromboxane (TxA2), nitric oxide (NO), and K+ channels. Intravital microscopy was used to measure vasodilation responses to acetylcholine in cremasteric arterioles of mice 11 wks after injection with mock inoculum or CMV. A PGI2 synthase inhibitor impaired dilation in mock, but not CMV, mice. Inhibition of TxA2 synthase had no effect in mocks, but partly restored dilation in CMV mice. A TxA2 receptor antagonist behaved similarly. This suggests that COX‐2‐PGI2 responses are impaired by CMV via COX‐1‐dependent TxA2 generation. NO synthase inhibition reduced dilation more significantly in mock vs. CMV mice. This implicates reduced NO bioavailability in CMV‐induced dysfunction at 11wk. Dilation was similarly impaired in mock and CMV groups by high [KCl], suggesting that CMV does not alter endothelium‐dependent hyperpolarization. Our data demonstrates that persistent CMV infection leads to arteriolar dysfunction by disrupting PGI2‐ and NO‐dependent vasodilation.
Role of CYP4A/20‐HETE Pathway in Vascular Oxidative Stress in the Dahl Salt‐Sensitive Rat
Authors: Lukaszewicz Kathleen , Falck John , Lombard Julian
Published: June 2021

We tested the hypothesis that cytochrome P450‐4A (CYP4A) and 20‐hydroxyeicosatetraenoic acid (20‐HETE) contribute to vascular oxidant stress and endothelial dysfunction in Dahl salt‐sensitive (SS) rats. Endothelial dysfunction in SS rats was ameliorated by a CYP4A inhibitor (DDMS) or a 20‐HETE antagonist (20‐HEDE), and in SS‐5BN consomic rats carrying CYP4A genes from the Brown Norway rat on the SS genetic background. Superoxide (SOX) levels, assessed via DHE fluorescence, were significantly higher in cerebral arteries of SS rats vs. SS‐5BN rats fed either normal salt (NS, 0.4% NaCl) or high salt (HS, 4.0% NaCl) diet. DDMS reduced SOX in arteries of HS‐fed SS rats to the same degree as a superoxide scavenger (PEG‐SOD), suggesting that the CYP4A/20‐HETE system contributes to elevated vascular SOX in HS‐fed SS rats. CYP4A protein expression was significantly higher in cerebral arteries of SS rats fed NS or HS diet vs. SS‐5BN rats. CuZn SOD, MnSOD, and EC‐SOD expression was similar in SS vs. SS‐5BN, indicating that the elevated vascular SOX in SS vs. SS‐5BN rats is not due to differences in antioxidant defenses. Treatment with the superoxide scavenger tempol had no effect on CYP4A expression in either strain. These findings indicate that increased expression/activity of the CYP4A/20‐HETE system contributes to vascular oxidant stress and endothelial dysfunction in SS rats fed either NS or HS diet. [NIH #HL65289, HL72920, HL92026, and AHA].
In hypertension CYP450A metabolite 20‐HETE exacerbates flow‐induced arteriolar constriction and promotes cerebrovascular inflammation
Authors: Toth Peter , Csiszar Anna , Sosnowska Danuta , Sonntag William E. , Ungvari Zoltan , Koller Akos
Published: June 2021

Hypertension impairs local regulation of cerebral blood flow by increasing vascular tone and inducing cerebrovascular inflammation thereby contributing to the pathogenesis of ischemic cerebrovascular diseases (i.e. stroke, vascular cognitive impairment). To test the hypothesis that 20‐hydroxyeicosatetraenic acid (20‐HETE) contributes to hypertension‐induced cerebrovascular alterations, we treated spontaneously hypertensive rats (SHR) with HET0016, an inhibitor of 20‐HETE synthesis. Although HET0016 decreased systolic blood pressure, the treated animals remained hypertensive. In cerebral arteries isolated from HET0016‐treated SHRs flow‐induced constriction was attenuated and endothelial dilation to acetylcholine was improved. HET0016 treatment inhibited cerebrovascular inflammation in SHRs as shown by the reduced expression of pro‐inflammatory cytokines TNF, IL‐1 and IL‐6. Taken together, in hypertension increased production of 20‐HETE contributes to cerebromicrovascular inflammation and dysfunction, which likely promote the pathogenesis of ischemic cerebrovascular diseases.

Supported: AHA, Founders Aff., 0855910D, NIH PO‐1 HL‐43023, Hungarian Natl. Sci. Res. Fund (OTKA‐K71591, K67984), MHT 2011.