bluuebunny/crossref_metadata_2025 huggingface dataset:
Recruitment in a pediatric clinical research trial targeting underserved populations: efforts and challenges (828.9)
Authors: Ireland Kathy , Manders Aaron , Corkey Barbara , Lenders Carine
Published: June 2021

Purpose: To describe recruitment difficulties in a pediatric clinical research trial targeting underserved populations.

Methods: We planned a 6‐month randomized clinical trial of glutamine and placebo to reduce HOMA_IR and weight gain in obese 12‐19 yo adolescents. An estimated 264 patients had to be screened to recruit 66 with insulin levels 蠅15 microU/L. Participants were asked to come to 5 visits at the General Clinical Research Center (GCRC) for 2‐3 hrs at 8am and received cash incentives at each visit. Increased recruitment efforts were put in place with the help of trained student volunteers for 14 weeks (6/10‐9/10). Subjects were recruited from BMC’s pediatric outpatient clinics including the pediatric obesity program which was staffed by members of the research team.

Results: 2002 adolescents had scheduled clinic visits at BMC. 546 met initial BMI and age criteria. After further exclusions, 179 were eligible; of which 4 made an appointment for a screening visit (SV) and 175 were either not approached, unavailable, refused to participate, or did not follow‐up. Additionally, 120 letters were sent to patients from the BMC obesity clinic, 4 attended a SV. Of the 8 adolescents that attended the SV, 7 were from BMC’s obesity clinic. 2 enrolled while 6 were ineligible due to insulin levels (n=4), medications, and payment issues. Both enrolled subjects were lost to follow‐up.

Discussion: Recruitment from the clinic where researchers had an existing relationship with patients was most successful. Challenges included strict inclusion criteria and early appointments in a new setting. Future trials should use looser criteria and methods to make participation less burdensome.

Grant Funding Source: Supported by NIH NDKxxxxx
The protective effect of Moringa oleifera leaf extract on fructose‐mediated protein glycation in vitro (829.10)
Authors: Ngamukote Sathaporn , Nunthanawanich Pornpimon , Adisakwattana Sirichai , Mäkynen Kittana , Sapwarobol suwimol , Dahlan Winai
Published: June 2021

Background: Chronic hyperglycemia in diabetes causes a non‐enzymatic glycation between reducing sugars and amino groups of proteins, resulting in production of advanced glycation end products (AGEs). It has been known that AGEs play a pivotal role in the pathogenesis of both microvascular and macrovascular complications of diabetic patients. Moringa Oleifera is one of the well‐known Thai medicinal plants that have been shown to have the favorable effects in the treatment and prevention of diabetes through various mechanisms. However, the antiglycation activity of Moringa oleifera leaf extract has not been investigated. Objective: The purpose of this study was to determine the inhibitory effect of Moringa oleifera leaf extract (MOE) on fructose‐induced protein glycation. Methods: The various concentrations of MOE were incubated with bovine serum albumin (BSA) and 0.5M fructose at 37ºC for 7, 14, 21 and 28 days. Fluorescence AGEs formation, fructosamine and β‐amyloid structure level were measured. Results: MOE (0.5‐2.00 mg/ml) showed a dose‐dependent decrease in fluorescence AGEs formation. Moreover, MOE significantly reduced fructosamine and β‐amyloid structure level when compared to the negative control. Discussion and Conclusion: MOE may reduce diabetic complications through inhibiting the AGEs formation, decreasing fructosamine level and also preventing protein aggregation (β‐amyloid structure). Additionally, this research may also provide beneficial effects of MOE on reducing the risk of advanced glycation endproduct associated diseases.

Grant Funding Source: Supported by Thailand Research Fund
Low intake of dietary antioxidants is associated with oxidative stress in Omani adults newly diagnosed with type 2 diabetes (829.16)
Authors: Waly Mostafa
Published: June 2021

Background: Recent epidemiological studies strongly indicate that low physical activity and a diet low in vegetables and fruits increased risk of type 2 diabetes (T2DM) among adults. Environmental insults induce oxidative stress that has been suggested to occur in the context T2DM pathology. The prevalence of T2DM in the Omani adults’ population is continuously increasing and has been linked to obesity and a sedentary life style. Although the protective role of dietary antioxidants in the pathogenesis of T2DM and oxidative stress has been investigated in Western populations, no study has been carried out in Omani adults with T2DM. Objective: The purpose of this study was to examine the status of dietary antioxidants and oxidative stress Omani adults newly diagnosed with T2DM. Methods: A case‐control study involved 200 Omani adults (100 patients and 100 age‐ and gender‐matched healthy controls). Several parameters were investigated, including dietary intake, physical activity level and biochemical assessments of dietary antioxidants and oxidative stress markers. Results: Among the studied subjects; there was no significant difference in the physical activity level, meanwhile low serum levels of dietary antioxidants, and oxidative stress indices were prevalent in patients with T2DM compared with controls. The oxidative stress insults levels were higher in subjects living in urban areas versus rural areas. Conclusion: Low intake of dietary antioxidants is associated with oxidative stress and collectively increasing the risk of T2DM among high risk groups of the Omani adults’ population.
Chrysin inhibits diabetes‐associated renal fibrosis through blocking epithelial to mesenchymal transition in tubular epithelial cells under high glucose episode (829.17)
Authors: Kang Min‐Kyung , Kang Young‐Hee
Published: June 2021

Renal fibrosis is a crucial process in the pathogenesis of diabetic nephropathy (DN). This process, known as epithelial to mesenchymal transition (EMT), contributes to the renal accumulation of matrix proteins, where renal tubular epithelial cells play an important role in progressive renal fibrosis. The current study investigated that chrysin (5,7‐dihydroxyflavone), a flavonoid present in bee propolis and herbs, inhibited renal EMT and fibrosis followed by chronic hyperglycemia. Human renal proximal epithelial cells were exposed to 5.5 mM or 33 mM glucose in the absence and presence 1‐20 μM chrysin for 72 h. Mannitol was employed as an osmotic control. Chrysin significantly inhibited renal EMT process through blocking high glucose (HG)‐induced expression of the mesenchymal markers vimentin and α‐smooth muscle actin in renal tubular cells. HG enhanced E‐cadherin expression in tubular cells and retarded N‐cadherin induction, which was reversed by chrysin. In addition, chrysin effectively deterred the excessive deposition of extracellular matrix components of collagen IV and connective tissue growth factor by HG, indicating the blockade of renal fibrosis resulting from myofibroblasts after HG episode. These results demonstrate that chrysin inhibits EMT‐ mediated renal tubular fibrosis due to chronic hyperglycemia. Therefore, chrysin may be a potent renoprotective agent for the treatment of DN.
Efficacy of consumption of whole and fractionated pulses on plasma lipids in diet‐induced dyslipidemic Syrian hamsters (829.19)
Authors: Ramprasath Vanu , Omar Jaclyn , Sihag Jyoti , Yang Shuo , Ames Nancy , Jones Peter
Published: June 2021

The objective was to examine effects of whole and fractions of pinto beans and peas on diet induced dyslipidemia. Hamsters were randomly assigned to consume high fat diet (n=12 each) including control (CON); bean starch (BS); bean fibre (BF); whole bean (WB); cooked whole bean (CWB); inner pea fibre (IPF); pea hull fibre (PHF); pea protein isolate (PPI); pea protein concentrate (PPC); or whole pea (WP) for 4 wk. Reduced feed intake (FI) and body weight gain (BWG) were observed with WB (FI: p=0.030; BWG: p<0.001), BS (FI: p=0.041; BWG: p<0.001) and BF (FI: p=0.037; BWG: p<0.001) compared to CON. However, both FI and BWG did not differ between CWB and CON. Plasma total, HDL and non‐HDL cholesterol, triglyceride and glucose concentrations were lower (p<0.05) in CWB, WB, BS and BF compared to CON. Furthermore, no changes in plasma lipids were observed with BF and BS compared to WB. No changes in FI, BWG and plasma glucose and lipid profile were observed with any of the peas supplemented groups compared to CON. A positive hemagglutination was observed with raw WB, BS and BF, but not CWB, against rabbit erythrocytes. Data suggest that, while whole and fractionated peas failed to alter plasma lipids and glucose levels, fractions and whole beans possess lipid lowering and hypoglycemic effects in hamsters, but must be cooked prior to intake. Results indicate that bean consumption can improve blood lipids and reduce cardiovascular disease risk

Grant Funding Source: Supported by Pulse Canada; Agriculture and Agri‐Food Canada
(‐)‐Epicatechin: cardiometabolic clinical effects supported by preclinical model (829.2)
Authors: Gutierrez‐Salmean Gabriela , Ortiz Pilar , Ramirez Israel , Meaney Eduardo , Villarreal Francisco , Ceballos Guillermo
Published: June 2021

Background. Obesity associates with cardiometabolic disruptions. Lifestyle changes and pharmacologic therapies show moderately effective results. (‐)‐Epicatechin (EPI), the most abundant flavanol in cacao, has been associated with better cardiometabolic health.

Objective. To assess the effect of EPI in humans and explain findings through a preclinical study.

Methods. In humans, we performed a single‐dose of EPI oral metabolic tolerance test and a 1‐week trial, measuring cardiometabolic endpoints. In rats, we induced weight gain and cardiometabolic disruptions by a high‐fat diet; afterwards, EPI was daily administered for 15 days. Weight gain, glycemia, arterial pressure, triglyceridemia, and HDL‐cholesterol were measured; also, immonublot in skeletal muscle and adipose tissue were performed.

Results. EPI enhanced lipid oxidation and attenuated hyperglycemia and hypertriglyceridemia during postprandial metabolism. EPI induced weight‐loss, fat loss, and improved cardiometabolic endpoints in humans. In rats, EPI reduced weight gain, blood triglycerides, and hyperglycemia. EPI increased the expression of sirtuins, PCG‐1α, mitofilin, UCP.

Conclusions. EPI improved several cardiometabolic risk factors in both humans and animals by modulating energy metabolism. These findings make EPI an attractive candidate for human use in regards of treating obesity and its associated comorbidities.
Grapeseed polyphenol + ∂‐tocotrienol favorably alters lipid profile in C57BL/6J mice with diet‐ induced non‐alcoholic steatohepatitis (829.28)
Authors: Mendez Melissa , Vijayagopal Parakat , Juma Shanil , Prasad Chandan , Imrhan Vicky
Published: June 2021

Diets high in fat are associated with the development of obesity and liver pathologies such as nonalcoholic fatty liver disease (NAFLD) and nonalcoholic steatohepatitis (NASH). Currently there are no safe drug therapies for NAFLD or NASH, therefore there is a need for alternative therapies. Studies have shown that grapeseed polyphenol (GSP) and ∂‐tocotrienol (∂‐T) may have benefits in reducing NASH‐like symptoms. In this study we have examined the effects of dietary GSP, ∂‐T, and their combination (GSP+ ∂‐T) in C57BL/6J mice fed a high calorie high fat diet for 20 weeks to induce NASH‐like symptoms. The control group was fed high fat diet throughout the study. Hepatic and serum total cholesterol TC), triglycerides (TG), and vitamin E concentrations along with serum free fatty acids (FFA) were assayed. Hepatic and serum TC were significantly lower (p<0.05) in GSP and GSP+ ∂‐T compared controls and ∂‐T groups whereas hepatic TG was lower (p<0.05) only in GSP+ ∂‐T group. Both hepatic and serum vitamin E levels were lower (p<0.05) in GSP+ ∂‐T compared to control and ∂‐T groups. Serum TG and FFA were similar among the groups. Dietary GSP supplementation alone and in combination with ∂‐T attenuates NASH‐like features in C57Bl/6J mice by reducing serum and hepatic lipids.

Grant Funding Source: Supported by the Research Enhancement Program, Texas Woman’s University.
Dietary garcinol retarded pancreatic cancer progression in kras and p53 transgenic mouse model (829.29)
Authors: Saadat Nadia , Goja Arvind , Gupta Smiti
Published: June 2021

Pancreatic adenocarcinoma has less than 6% five year survival rate, the lowest in all cancer types. Garcinol is a polyisoprenylated benzophenone derivative from Garcinia Indica fruit extract. Invitro data from our lab showed that Garcinol displayed a dose dependent favorable response in pancreatic cancer lines. In this study, we investigated the invivo effects of dietary Garcinol in a transgenic mouse model of Pancreatic Cancer. Reduced tumor progression observed in MRI scans in live animal was investigated through changes in micro RNA and mRNA expressions (microarray& RTPCR). We probed the differences in Urinary Metabolomic profiles (MP) of Garcinol treated mice (KGr) compared to non‐treated (KC) mice for metabolic response to the treatment. Nuclear Magnetic resonance spectroscopy was performed on Varian‐500S. Simca P+ and CHENOMX NMR suite were used for Metabolomic analysis of urinary spectra. Urinary MP from week 6 KGr group were closer to non‐ cancer (CC) and week 2 of KC group. Target analysis with CHENOMX identified Taurine, Tartrate and Phenylacetate, found to be in higher concentrations in KGr group compared to KC group and Allantoin was found to be lower in KGr group compared to KC group. Garcinol has shown pleotropic effects, targeting multiple tumor related pathways. Further investigation of molecular pathways identified can lead to its incorporation as a chemotherapeutic agent in a clinical trial.
Flavonoid luteolin supplementation inhibits diethylnitrosamine‐initiated alcohol‐promoted hepatic inflammation and precancerous lesions in mice (829.7)
Authors: Rafacho Bruna Paola , Stice Camilla , Liu Chun , Ausman Lynne , Greenberg Andrew , Wang Xiang‐Dong
Published: June 2021

Chronic and excessive alcohol consumption is an established risk for hepatic inflammation and carcinogenesis. Luteolin is one of the most common flavonoids present in plants and has potential beneficial effects against chronic diseases. In this study, we examined the effect and potential mechanisms of luteolin supplementation in a carcinogen (diethylnitrosamine [DEN])‐initiated alcohol‐promoted pre‐neoplastic liver lesion mouse model. C57BL/6 mice were injected with DEN (i.p. 25 mg/kg of body weigth [BW]) at 14 days of age. At 8 weeks of age mice were group pair‐fed with Lieber‐DeCarli liquid control diet or alcoholic diet (EtOH diet, 27% total energy from ethanol) and supplemented with a dose of 30 mg luteolin/kg BW/day for a treatment period of 21 days. Results show a significant induction in the incidence and multiplicity of basophilic and eosinophilic foci, markers of pre‐neoplastic liver lesions, in DEN‐injected mice fed EtOH diet. The presence of pre‐neoplastic lesions was significantly correlated with the induction of steatosis and inflammatory foci in these mice. Supplementation with luteolin inhibited the presence of pre‐neoplastic liver lesions in DEN‐injected mice fed EtOH diet. Dietary luteolin significantly reduced the severity of hepatic inflammatory foci with an associated non‐significant reduction in expression of hepatic inflammatory cytokines TNF‐α, IL‐6 and IL‐1β. There was no difference on hepatic protein levels of sirtuin 1 (SIRT1), however, luteolin supplementation significantly reversed alcohol‐reduced SIRT1 activity assessed by the ratio of acetylated and total FoXO1. The study demonstrated that luteolin prevents EtOH diet‐promoted hepatic inflammation and pre‐neoplastic lesion, potentially mediated by SIRT1‐FoxO1 signaling pathway.

Grant Funding Source: USDA/ARS grant 1950‐51000‐074S
Processing and storage of ratite oils affects radical scavenging ability and primary oxidation (830.15)
Authors: Mashtoub Suzanne , Tran Cuong , Howarth Gordon
Published: June 2021

BACKGROUND. Treatments for cardiovascular and gastrointestinal disease seek to minimise oxidative damage using antioxidants. Ratite (flightless birds) oils predominantly comprise fatty acids (FA) varying in composition between species. AIM. We investigated the influence of rendering and storage methods on radical scavenging activity (RSA) and primary oxidation status of ratite oils. METHODS. Emu Oil (EO1‐7), Ostrich Oil (OsO), Rhea Oil (RO) and Olive Oil (OlO) were tested for free RSA against 2,2‐diphenyl‐1‐picryl hydracyl (1/IC50 g/ml) and primary oxidation (peroxide mEqO2/Kg). p<0.001 was considered significant. RESULTS. RSA (g/ml) of EO1 (558±22) was greater than EO2 (8±0.6), EO5 (413±26), EO6 (16±0.3), EO7 (2±0.2), OsO (313±12), RO (32±12) and OlO (196±4), but less than EO3 (717±32; p<0.001). RSA of OsO was more pronounced than RO (p<0.001). Primary oxidation (mEqO2/Kg) of EO1 (97±0.6) was greater than EO2 (28±0.2), EO5 (11±0.6), OsO (50±0.9) and OlO (61±0.9). CONCLUSIONS. Emu Oil conferred the greatest radical scavenging activity compared to Ostrich and Rhea Oil, possibly attributed to its high UFA:SFA ratio and non‐triglyceride fraction minor constituents. Rendering and storage affected radical scavenging activity and primary oxidation of Emu Oil. Regulatory control of these parameters could minimise differences in therapeutic efficacy of ratite oils.
Effects of edible mushrooms (Pleurotus ostreatus (Jacq.) P. Kumm., Pleurotus eryngil, Flammulina velutipes) extracts on immune cell activation in mice (830.17)
Authors: Ryu Hye‐sook , Kim Kyoung‐ok , Liu Yanan , Yoon Leena , Kim Hyun‐sook
Published: June 2021

Oyster mushroom (Pleurotus ostreatus (Jacq.) P. Kumm.), king oyster mushroom (Pleurotus eryngil), and winter mushroom (Flammulina velutipes) are typical edible mushrooms in many Asian countries. The splenocyte proliferations were measured at 48 hours after treatment of water extract those mushrooms in seven concentrations (0, 5, 10, 50, 100, 250, 500, and 1000 ug/ml). Also cytokine productions (TNF, IFN‐γ, IL‐2, IL‐4, IL‐5) in activated macrophage were measured. Splenocyte proliferation was enhanced when the 100, 250, 500, 1000 ug/ml oyster mushroom extracts were administered compared to control (0 ug/ml). Splenocyte proliferation were significantly higher in 50, 100, 250, 1000 ug/ml king oyster mushroom extract treated cells. Splenocyte proliferations of cells treated with winter mushroom extract were significantly increased in all groups except the group treated 5ug/ml extract. Th1 cytokine productions (TNF‐α and IFN‐γ) were increased by administration of mushroom extracts whereas levels of Th2 cytokines such as IL‐4 and IL‐5 did not change. These primary results suggest that oyster, king oyster, and winter mushrooms may be enhance the immune function in fairly high concentrations.
Skin whitening effects of flower extracts with high antioxidant activities (830.20)
Authors: Lee Bog‐Hieu , Choi Joo‐Yeon , Yu Miri , Jeong Gi Hee
Published: June 2021

This study was aimed to investigate whitening effect of seven flower extracts with high electron donating ability (EDA). CCK‐8 assay revealed that the cell viability rate of all the flower extracts was significantly higher than that of control. Whitening effects of the flower extracts were measured by mushroom tyrosinase inhibition rate, cellular tyrosinase activity and cellular melanin contents in B16F1 melanoma cells. Rhododendron molle (Blume) G. Don extract reduced mushroom tyrosinase activity by 35.1% than the control and ascorbic acids. At 500 ppm, Aruncus dioicus var. kamtschaticus extract suppressed tyrosinase activity by 45.8%. In case of cellular melanin content, Euscaphis japonica extract was significantly lower than the control and the ascorbic acid. As for gene expression level of TRP‐1, TRP‐2, and tyrosinase, relative quantification value of the Euscaphis japonica extract showed inhibition of expression of three melanogenic enzymes. In conclusion, flower extracts had shown strong whitening effect. These findings suggest that flower extracts may can be further developed and potential skin‐whitening or anti‐melanogenesis preparation for cosmetics and therapeutic use.

Grant Funding Source: supported by Basic Science Research Program through the Natiional Research Foundation of Korea(NRF)
Determination of plasma and leukocyte ascorbate concentrations in a randomized, double‐blind trial (830.3)
Authors: Mitmesser Susan , Combs Maile , Evans Mal
Published: June 2021

Plasma ascorbate reflects recent intake, whereas leukocyte ascorbate more closely reflects cellular stores and total body pool.

Objective was to evaluate the bioavailability of ascorbate in plasma and leukocytes from a novel vitamin C formulation.

In this double‐blind, cross‐over, males and females between 18‐60 years were randomized to receive one of the following: Placebo (P, 0mg vitamin C), Ascorbic Acid (AA, 1000mg vitamin C) or Ester‐C® (EC, 1000mg vitamin C + threonate and furanone) as a single dosage. Plasma and leukocyte ascorbate were measured at 2, 4, 8, and 24 hours following treatment.

An intention‐to‐treat analysis was conducted on thirty six subjects (18 males, 18 females). Plasma ascorbate percent change from Baseline was significantly higher for EC and AA at 2 hours compared to P (EC=135.55±176, AA=126.60±135, P=2.35±33; p<0.001). At 4, 8, and 24 hours the percent change from Baseline was only significantly higher for EC compared to P (all p蠄0.035). The percent change in leukocyte ascorbate was significantly higher from Baseline for EC at 2, 4, 8, and 24 hours (all p蠄0.041) and P at 4 and 24 hours (p蠄0.035) but there was no change from Baseline for the AA group.

Vitamin C from Ester‐C and Ascorbic Acid has greater plasma bioavailability than Placebo. Compared to Baseline, Ester‐C® provides a higher percent change in leukocyte ascorbate, which may be due to the presence of threonate and furanone in the formula.
Sasa quelpaertensis leaf extract suppresses inflammation in DSS‐induced colitis in vivo (830.7)
Authors: Kim Kyung Mi , Kim Yoo‐Sun , Lim Ji Ye , Min Soo Jin , Ko Hee‐Chul , Kim Se‐Jae , Kim Yuri
Published: June 2021

Sasa quelpaertensis Nakai is a species of bamboo grasses grown in Mt.Halla on Jeju Island in South Korea. Sasa leaves have been suggested to have anti‐diabetic, anti‐cancer, and anti‐inflammatory effects. Inflammatory bowel disease (IBD) is a disease characterized by chronic inflammatory disorders of the gastrointestinal tract. The aim of this study is to investigate the possible protective role of Sasa quelpaertensis leaf extract (SQE) on dextran sodium sulphate (DSS)‐induced colitis in mice model. We found that SQE attenuated the severity of IBD assessed by disease activity index (DAI) score, shrinkage of colon length, and histopathological changes. SQE also significantly suppressed the DSS‐induced serum inflammatory cytokines including TNF‐α, IL‐1β and IL‐6. Furthermore, SQE down‐regulates the expression of TNF‐ α, COX‐2, iNOS, NF‐kB, and PPARγ. Furthermore, SQE down‐regulated ERK1/2 phosphorylation, but not p38 or JNK. These results suggest that SQE may a protective role in gut inflammation including IBD via regulation of inflammatory mediators and inflammation‐mediated signal pathway.

Grant Funding Source: Support by Rural Development Administration (# PJ009777) & brain korea 21 plus (# 22A20130012143)
Effect of grape consumption on metabolic parameters in obese adults (831.11)
Authors: Giebler Kathryn , Sato Jennifer , Stevens Allison , Ford Sarah , Steinberg Francene
Published: June 2021

Consumption of polyphenol‐rich grapes has previously shown cardioprotective effects. This randomized, double‐blind, crossover study aimed to determine the impact of whole grapes on biomarkers of cardiovascular disease risk compared to placebo. We examined plasma lipids, glucose, insulin, and C‐reactive protein in obese adults (mean BMI 39.2 ± 11.7) in good health. Diets included 60 g of whole grape powder (WGP), equivalent to 2 cups whole grapes providing 300 mg total polyphenols, or placebo(C) matched in calories and appearance providing 0 mg polyphenols. Subjects completed two 4‐wk intervention periods, separated by a 2‐wk washout, during which they consumed WGP or C once daily in addition to low polyphenol diets. Fasting blood samples were drawn at beginning and end of each intervention period. Acute postprandial effects at 1, 3, and 5 h were examined following consumption of WGP or C with a standardized high fat, high carbohydrate test meal. Preliminary data reveals subjects receiving WGP had no significant differences in fasting metabolic parameters compared to C, with exception of HDL (p<0.05). Similarly, postprandial AUC for glucose, insulin, and TG did not differ. Further evaluation is needed to determine if grape consumption improves other metabolic biomarkers in the obese population.

Grant Funding Source: Supported by CTGC and USDA CRIS CA‐S*‐NTR‐6316‐H
Polyphenols from açaí: a strategy to modulate inflammation and hypertension (831.2)
Authors: Daleprane Julio , Aguiar Cristiane , da Silva Renata Cristina , Rainho Rafaela , Moura Roberto , Resende Angela de Castro Resende
Published: June 2021

The aim of this study was evaluated the anti‐inflammatory effects in vivo model of renovascular hypertension (RH) and in endothelial cells (ECs). Rats were treated with ASE 200 mg.kg‐1.day (or vehicle) for 40 days. Oxidative damage, antioxidant activity and the expression of endothelial nitric oxide synthase (eNOS), superoxide dismutase 1 and 2 (SOD 1 and 2), metalloproteinase (MMP‐2) and tissue inhibitor of metalloproteinases (TIMP)‐1 were determined. Structural changes in the aortic and mesenteric MMP‐2 levels were evaluated. In in vitro experiments, ECs were treated with 100 ug/mL of ASE and the expressions of inflammatory markers were evaluated. ASE reduced the increased mesentery levels of malondialdehyde and carbonyl protein in hypertensive rats. SOD, catalase and glutathione peroxidase activities and SOD1 and 2, eNOS and TIMP‐1 expressions were reduced. ASE prevented the vascular remodeling, as well as, the increased MMP‐2 expression and levels found in hypertensive rats. In ECs ASE was able to decreased TNFα, eNOS and IL‐8 mRNA expression. These results suggest that ASE prevents the endothelial dysfunction and vascular structural changes associated with hypertension

Grant Funding Source: FAPERJ, CNPq and CAPES
Effect of orally administered EPA and DHA on congestive heart failure in rat (831.7)
Authors: Yamanushi Tomoko , Kabuto Hideaki , Hirakawa Eiichiro , Janjua Najma , Takayama Fusako , Mankura Mitsumasa
Published: June 2021

This study assessed the effect of EPA and DHA on congestive heart failure (CHF) in rat. Four‐week‐old male rats were divided into 4 groups: C (normal, administered saline, n=4‐5); H (with CHF, administered saline, n=10‐11); HE (with CHF, administered EPA‐ethyl ester (EPA‐Et), n=9‐10); and HD (with CHF, administered DHA‐ethyl ester (DHA‐Et), n=10). CHF was produced by subcutaneous injection of monocrotaline (MCT) into 5‐week‐old rats. Starting from 3 days before MCT injection, rats were orally administered (1g/kg body weight) saline or one of the two fatty acids daily, anaesthetized on day 24, their electrocardiograms (ECGs) recorded, serum samples drawn, and then killed. The heart was removed, and kept at ‐80ºC or fixed with paraformaldehyde. Fatty acid levels in the heart and triglycerides in serum were determined. Pathological assessment was performed on heart tissue. The Q and T waves (QT) interval on ECG was significantly shorter (P 蠄 0.05) in C and HD groups than in H and HE groups. In the heart, EPA level was higher in HE group and DHA level higher in C and HD groups, compared to H group (P 蠄 0.05). Triglyceride level was significantly decreased (P 蠄 0.05) in H, HE and HD groups compared to C group. In HD group, mononuclear cells tended to infiltrate less into myocytes than in H group (P = 0.06). The right ventricle weighed significantly more (P 蠄 0.05) in H, HE and HD groups compared to C group. The mean survival rate in the three experiments was 100% for C group, 74% for H group, 65% for HE group and 70% for HD group. Administration of EPA‐Et or DHA‐Et may affect cardiac function through alteration of heart fatty acid composition. Furthermore, DHA‐Et administration may contribute to ameliorating CHF.

Grant Funding Source: Supported by JSPS KAKENHI Grant Number 21500788
Effect of immune modulators on the activity of enzymes after experimental myocardial infarction (832.10)
Authors: Sarapultsev Alexey , Gette Irina , Abidov Musa , Sarapultsev Petr , Chupakhin Oleg , Danilova Irina
Published: June 2021

It was revealed (patent RUS 2437165 from 13.04.2010) that application of immune modulators fosters reducing the severity of inflammation reaction, faster cicatrization, provokes angiogenesis. As tissue regeneration depends on energy supply of the cell, the goal of the research was studying the effect of immune modulators on the activity of enzymes involved in energy exchange in the damaged myocardium. The experiment was conducted with 65 male rats of the Wistar line. The immune modulators used were derivatives of 1,3,4‐thiadiazine (L‐17) and 3‐aminophtalhydrazine (AMP), exerting antioxidant effect as well. Modeling myocardial infarction was verified by morphological research methods and increasing activity of organo‐specific myocardium enzymes in the blood plasma. With experimental myocardial infarction, the myocardium tissue showed CPK activation and decreasing activity of LDH1,2 isoenzymes. Introduction of AMP facilitated faster cardiac enzymes normalization in the blood plasma of the animals than introduction of L‐17. Nevertheless, both compounds prevented decreasing activity of LDH1,2 induced by myocardial infarction modeling in the myocardium at early stages (1 day), without decreasing CPK activity at all stages of the experiment.

Grant Funding Source: Supported by grant 12‐Ц‐41059
Targeted inhibition of ATP‐binding cassette transporter A1 protein degradation promotes reverse cholesterol transport and reduces atherosclerosis in mice (832.15)
Authors: Huang Linzhang , Zhu Haibo
Published: June 2021

Objective ATP‐Bing Cassette Transporter A1 (ABCA1) is a key protein in the rate‐limiting step of reverse cholesterol transport, which maintains plasma HDL levels. Transcriptional regulating ABCA1 promotes HDL generation and RCT is well established, however, little is known about the posttranspcriptional inhibition of ABCA1 degradation on macrophage RCT and atherosclerosis. We therefore investigate whether suppressing ABCA1 degradation would promote RCT and reduce atherosclerosis.

Methods and results Pulse‐chase analysis revealed that a small molecular (IMM‐H007) significantly inhibits ABCA1 degradation and facilitates its cell‐surface localization in THP‐1 macrophages. In vitro tests described that IMM‐H007 promote cholesterol efflux via ABCA1‐mediated pathway. Animals fed a western diet containing 1.25% cholesterol with IMM‐H007 treatment showed an increase in circulating HDL levels and enhanced RCT to the plasma, liver, and feces. Additional, elevated hepatic ABCA1 expression and functional HDL were also observed in mice after IMM‐H007 administration. Furthermore, suppressing ABCA1 degradation gave rise to the reduction of plaque size, lipid content and increase markers of plaque stability in apoE‐/‐ mice.

Conclusion Our findings reveal that raising HDL levels by inhibiting ABCA1 protein degradation enhances reverse cholesterol transport and decreases plaque formation, suggesting that it is a promising HDL therapeutic strategy to protect against atherosclerotic. IMM‐H007 could be a leading compound for development of drugs for raising HDL quantity and function.
Elderly individuals with high IL‐6 levels have proinflammatory HDL (832.16)
Authors: Farazmand Marya , Dadgostar Amir , Seddigh Farhad , Reuben David
Published: June 2021

Introduction: High density lipoprotein removes the oxidatively modified lipids and other toxic molecules from peripheral cells for elimination and thus it is anti inflammatory and protective. Under chronic acute phase pressure, unsaturated fatty acids and phospholipids in HDL undergo oxidative modification and that results in HDL losing its anti inflammatory and protective capacity. Objective: we sought to determine if HDL from individuals with inflammatory background can retain its protective ability. Methods: We recruited a group of elderly, visiting the UCLA clinics (n=40), who had normal (Group 1) or higher than normal plasma IL‐6 levels (Group 2). We followed the UCLA Medical Center IRB rules and instructions and analyzed their plasma sample for HDL anti inflammatory capacity. This was done using monocyte chemotactic activity toward human artery wall cells in co culture. Results: While the plasma HDL cholesterol levels of the two groups was similar (59.2+ 7.6 and 61+6.9 mg per dL respectively) with no significant difference, the HDL protective capacity of the group with high serum IL‐6 level was significantly lower than the group 1 who did not have high circulating IL‐6 levels (‐ 35% vs + 67% respectively, p <0.01). Conclusion: The data obtained indicates that in the elderly with high plasma IL‐6 levels the inflammatory pressure results in the loss of the protective capacity of HDL and this in turn moves the organism toward a vicious cycle. HDL protects lipids and other substances form oxidative modification. Los of HDL protective capacity increases the modification of lipids and other macromolecules in LDL, in cell membrane and basically throughout the organism with the pursuing undesirable structural and functional consequences.

Grant Funding Source: NIH
The role of endothelial nitric oxide synthase coupling status during acute hyperglycemia as determined by real‐time measurements of blood nitric oxide and hydrogen peroxide in rat (832.2)
Authors: Bertolet Matthew , Minni Michael , Galbreath Tyler , Patel Shan , Selko Robert , Barsotti Robert , Young Lindon , Chen Qian
Published: June 2021

Acute hyperglycemia impairs vascular dilatory function in normal subjects. Normally, vascular endothelial function depends on nitric oxide (NO) production from coupled eNOS in the presence of cofactor 5,6,7,8‐tetrahydrobiopterin (BH4). By contrast, 7,8‐dihydrobiopterin (BH2, oxidized form of BH4) and/or lack of L‐arginine (eNOS substrate) causes eNOS uncoupling to produce superoxide (SO), which can be quickly converted to hydrogen peroxide (H2O2). The role of eNOS coupling status in mediating acute hyperglycemia‐induced vascular dysfunction is not known. We simultaneously measured blood NO and H2O2 during the infusion of saline (control), or 20% glucose with or without BH4 (MW=314 g/mol, 6.5 mg/kg), or L‐arginine (MW=211 g/mol, 600 mg/kg), or BH2 (MW=239 g/mol, 4 mg/kg) by NO or H2O2 microsensors (100 µm, WPI Inc.) from the femoral veins of male Sprague‐Dawley rats. We found that acute hyperglycemia (200 mg/dL, n=6) significantly increased blood H2O2 and reduced blood NO levels compared to the saline group over a 3 hr. period (n=7, p<0.05). By contrast, BH4 (n=6), or L‐arginine (n=5‐6), not BH2 (n=6‐7), significantly reduced blood H2O2 and improved vascular NO levels (p<0.05). This study provides novel evidence indicating that eNOS uncoupling during acute hyperglycemia causes endothelial dysfunction and oxidative stress. Promotion of eNOS coupling may be beneficial to maintain normal vascular function.

Grant Funding Source: Supported by Center for Chronic Disorders of Aging and Department of Bio‐Medical Sciences at PCOM
Alteration of retinal vascular development by postnatal systemic lipopolysaccharide administration: a novel model of retinopathy of prematurity (832.6)
Authors: Hong Hye kyoung , Oh Joo youn , Park Ji Hyun , Park Ji Yeon , Park Kyu Hyung , Choi Chang Wong , Woo Se Joon
Published: June 2021

Purpose: ROP is the leading cause of blindness in the children. Inflammation is being highlighted as the major factor for the morbidity of prematurity. We intended to investigate the effect of neonatal systemic inflammation on the retinal angiogenesis in newborn rats and to show the role of inflammation in the retinopathy of prematurity.

Method: Systemic inflammation was induced by the pre‐ and post‐natal intraperitoneal injections of lipopolysaccharide (LPS) in neonatal rats. Enucleation and retinal separation was performed at 1 and 2 post‐natal weeks. Analyses on morphology of retinal vessels, recruited immune cells, change of inflammatory cytokines were assessed with immunofluorescence, FACS and RT‐PCR.

Result: Administration of LPS postnatlly, not prenatally, induced inflammation in the retina with recruitment of CD11c positive cells and increased expression of a set of inflammatory cytokine genes. Postnatal LPS administration also suppressed retinal vessel development and deranged the retinal vascular structures combined with abnormal vascular tuft formation that resembles features of retinopathy of prematurity.

Conclusion: Neonatal systemic inflammation alters the normal development of retinal vessels by inducing retinal inflammation. Our model of systemic inflammation‐induced abnormal development of retinal vasculature can be a novel experimental model of retinopathy of prematurity.
The feline placenta as a physiologic model of complex vascular development (832.7)
Authors: Huckle William , Frydman Galit , Martin Cassandra , Weerakoon Shaneke , Gudenschwager Kristobal
Published: June 2021

The development of the placenta throughout gestation is vital for the normal progress of fetal growth to term. The placental vasculature develops early in pregnancy, becoming eventually a complex network that brings the fetal and maternal circulations into close proximity, allowing efficient intervascular exchange of material without frank comingling. In accord with our ongoing interest in cellular signaling mechanisms that govern vascular development, repair, and pathologies, we are evaluating the utility of the feline placenta as a setting in which to characterize developmental gene expression programs, growth factors and receptors involved in the genesis of normal placental vasculature. Placental tissues were obtained from elective ovariohysterectomies conducted at local spay‐neuter clinics, and gestation stages were estimated by fetal crown‐rump length. Preliminary qPCR‐based gene expression studies indicate that multiple isoforms of VEGF and its major receptor variants are expressed in the placenta throughout pregnancy, with relative declines at the mRNA level later in gestation when the vascular network is fully formed. Despite differences between human and feline placental anatomy and pathologies, the feline model represents a fruitful setting in which to study complex development of a vasculature of clear biological importance.
Comparison of cynomolgus macques (Macaca fasicularis) basic coagulation paprameters based on genetic origin versus attributed sources (833.4)
Authors: Waller Donald , McGeehan Elizabeth , Dubach Jean , Sinacore James
Published: June 2021

Cynomolgus macaques (Macaca fascicularis) are increasingly being used for a wide variety of studies, including anticoagulant therapy development and toxicology testing. Cynomoglus macaques are sourced from several different countries, some have indigenous populations and some have established breeding colonies from unknown indigenous populations. Genetic monitoring can verify the source of the macaques but is not usually included in study group monitoring. Small study group numbers and heterogeneous groups of animals can create difficulty in statistical interpretation of study results. The present study compares coagulation responses from animals attributed to sources in Vietnam, Cambodia, Philippines, Indonesia, China and Mauritius. Genetic characterization of 12 microsatellite loci used a Beckman/Coulter capillary electrophoresis system to determine genotypes. The baseline clotting profile was evaluated by determination of prothrombin time (PT), activated partial thromboplastin time (aPTT), common pathway (Heptest®), and thrombin time (TT). Blood was drawn and stored in EDTA for genetic analysis or immediately transferred to sodium citrate tubes, centrifuged to obtain plasma and stored at ‐80oC. Clotting tests were performed using the ACL 300+ and BBL Fibro System manual fibrometers. Statistical analysis for heterogeneity were performed with the groups of animals by their attributed source and reanalyzed after regrouping by genetic data. The heterogeneity was highly significant when the groups were evaluated based only on their attributed source, but was not significant when assigned to groups based on genetic characterization. The animals sourced from Mauritius were consistently more homogenous when compared to the other sources.

A clear understanding of the genetic background can insure group consistency and is essential when applying specific statistical tests to interpret data.
Differential restoration of prothrombin time / INR in plasma from warfarin‐treated patients by various prothrombin complex concentrates (833.6)
Authors: Abro Schuharazad , Hoppensteadt Debra , Eshraghi Reza , Only Arthur , Walenga Jeanine , Fareed Jawed
Published: June 2021

Introduction ‐ Conventional oral anticoagulant drugs produce their effects by suppressing the synthesis of functional forms of clotting factors II, VII, IX and X. Excessive inhibition of these factors results in bleeding complications. Prothrombin complex concetrates (PCCs) such as the Beriplex, Profilnine, and Feiba can be used to neutralize these effects. The purpose of this study was to compare the relative restoration of prothrombin time / INR in plasma samples collected from warfarin treated patients.

Materials and Methods ‐ Various PCCs were purchased and diluted at a working concentration of 10 U/ml. The plasma samples from patients treated with warfarin (n=15, PT 15‐40 sec) were supplemented with each of the different PCCs at a final concentration of 1U/ml. Saline supplemented samples from 10 individuals served as control. Prothrombin time / INR were measured using conventional methods. All results were expressed in terms of mean + 1SD and relative neutralization ratio was calculated.

Results ‐ The mean prothrombin time of the oral anticoagulated patients was 24.3 + 6.4 sec. Supplementation of Beriplex results in a total neutralization of this parameter with a mean of 11.5 + 0.7 sec. Profilnine partially neutralized with a value of 16.7 + 2.9 sec. Feiba produced a strong effect and shortened the prothrombin time to 8.9 + 0.4 sec.

Discussion ‐ These results suggest that Beriplex is capable of restoring the coagulant activity of the plasma samples from orally anticoagulated patients in a more efficient manner than Profilnine. Feiba produced a stronger response due to the presence of activated clotting factors.
Facilitation of urokinase‐mediated fibrinolysis by MST‐188 (833.7)
Authors: Fareed Jawed , Syed Daneyal , Emanuele Marty , Hoppensteadt Debra , Cho Jae
Published: June 2021

Introduction MST‐188 (Mast Therapeutics, San Diego, CA.) is a highly purified form of the , triblock copolymer poloxamer 188. It exhibits potent rheologic properties that result in improved blood flow especially under low shear conditions. Initial observations suggested that endogenous fibrinolysis is facilitated by this agent. In this study we investigate the effect of MST‐188 on urokinase induced fibrinolysis.

Materials and Methods A dynamic fibrinokinetic assay was used to assess both the assembly and dissolution of fibrin clots. MST‐188 (0 ‐ 10 mg/ml) was added to citrated human plasma with and without urokinase at a concentrations between 0 ‐ 2,500 U/ml (N=10‐15 donors). Clotting was initiated with the sequential addition of and calcium chloride (0.25 uM) and then thrombin (0.5u/ml). Absorbance was monitored to detect the assembly and dissolution of fibrin.

Results Control studies of MST‐188 alone had a dose dependent effect on fibrin assembly as reflected by both the rate of change and peak optical density. Urokinase alone showed no effect on clot formation but a dose dependent increase in clot lysis. Supplementation of MST‐188 to urokinase facilitated its lytic profile in a concentration dependent manner accelerating the time to onset of lysis and reducing the overall AUC for clot dissolution.

Discussion These results demonstrate that MST‐ 188 increases urokinase mediated fibrinolysis. The increase in the susceptibility of the formed clot to lysis in the presence of MST‐188 may be due to structural modification of the clot. MST‐188 may bind to fibrin strands, thereby modifying their accessibility to the lytic action of generated plasmin.
Analysis of smoking prevalence and smoking‐related diseases in the Estonian Genome Bank data (834.10)
Authors: Koks Gea , Koks Sulev , Raag Mait , Fischer Krista , Parna Kersti
Published: June 2021

Estonian Genome Bank is a population based biobank what combines DNA samples with general health questionnaire data. This repository contains data of more than 50,000 people. Database involves general data about the donors lifestyle (smoking, dinking, education, jobs etc) and about their health. We included to our analysis only these persons who responded about their smoking habits. Smoking status was divided into three categories: current smoker, former smoker and persons never smoked. Half of smokers have smoking duration over 20 years. Health status describes the disease people have. Our analysis focused on the smoking behaviour and its relations to the heart and lung disease and to the malignancies. Preliminary data analysis indicated that former smokers had higher prevalence of COPD and lung cancer than current smokers and never smokers. This finding may reflect that former smoker quit smoking due to emerging health problem.
Potential anti‐inflammatory effects of nebulized local anesthetics on inhaled chlorine lung injury (834.8)
Authors: Clodfelder‐Miller Buffie , Dewitte Carey , Xu Shin , Doran Steve , Matalon Sadis , Ness T
Published: June 2021

Chlorine (Cl2) is a chemical used in both industry and society alike, despite it being a highly irritant and reactive gas. Acute exposure can result in symptoms of airway obstruction such as wheezing, shortness of breath, mouth/nose/chest pain, and bronchospasm. Currently, there is no known antidote for chlorine’s poisoning effects. Potential treatments include topical exposure of the airways to nebulized drugs such as local anesthetics which have been shown to improve the mobility of monitored animals after exposure to Cl2 gas. In this research project, we assess the effects of the nebulized local anesthetics lidocaine (1% and 4%), 3% 2‐Chloroprocaine following exposure to 100 or 400 ppm of chlorine gas on biochemical measurements of injury to airway. Wet‐to‐dry lung weight ratios, bronchoalveolar lavage (BAL) cell counts and total protein, and lung homogenate cytokine levels were determined 24 hours after Cl2 gas exposure and anesthetic treatment. In mice exposed to 400 ppm Cl2 gas, 1% lidocaine had the greatest decrease of wet‐to‐dry lung weight ratio in addition to lower live cell infiltration and lower protein concentrations in BAL fluids indicating a potential anti‐inflammatory affect. Cytokine levels from lung homogenates after exposure and treatments had a decreasing trend compared to untreated animals. Levels of IL‐10, IFNγ, MCP‐1 and IL‐13 were significantly lower in the 1% lidocaine animal group. Similar but not identical results were found following treatment with 100ppm Cl2. Treatment with 4% lidocaine and 3% 2‐chloroprocaine had potential toxic effects. This study indicates that nebulized 1% lidocaine is potentially a good treatment option after chlorine induced lung injury based on behavioral and biochemical data.

Grant Funding Source: Supported by 1R21ES022876