bluuebunny/crossref_metadata_2025 huggingface dataset:
MZF‐1 regulates the CTGF gene in the hematopoietic compartment
Authors: Piszczatowski Richard
Published: June 2021

Connective Tissue Growth Factor (CTGF), an extracellular protein with key roles in wound healing and tissue homeostasis, has recently been shown to be in abundance in thrombocytes. Paradoxically, megakaryocytes, the platelet precursors in the bone marrow, contain no detectable levels of CTGF, and the source of CTGF in platelets is yet to be determined. It has been proposed that cells in the vicinity of the megakaryocyte secrete CTGF upon cellular signaling, at which point it is then internalized by fragmenting platelets entering the vasculature.

In this study, we report the discovery of a possible functional binding site for Myeloid Zinc Finger‐1 (MZF‐1), a transcription factor localized in the bone marrow, in the promoter region of the human CTGF gene, using bioinformatics. We further show that, in HeLa, HEK‐293, and select hematopoietic cells, binding of MZF‐1 to the CTGF promoter can be detected with Chromatin Immunoprecipitation (ChIP). Furthermore, we investigated the induction of CTGF by calcitriol (1,25‐ dihydroxyvitamine D3) via MZF‐1, through the PI‐3 kinase pathway in HS‐5 stromal fibroblasts, a possible candidate for the supply of CTGF to platelets entering the bloodstream. Finally, expression of CTGF is attenuated following ablation of MZF1 by miRNA. Taken together, our results argue that MZF‐1 is a key regulator of CTGF in the hematopoietic compartment, and may be a factor in the process of CTGF acquisition by thrombocytes.
Safer, brighter, two color fluorescent HRP‐ELISA systems
Authors: Harlan Fiona Karen , Paluck Tom C , Naleway John J
Published: June 2021

Quantification of HRP is a valuable tool for many applications including IgG detection for rapid screening of hybridoma culture media or ascites. ELISAs are commonly used for this purpose with Horse Radish Peroxidase (HRP) conjugated antibodies. This system while effective, has limitations stemming from the commonly used HRP substrate 3,3′,5,5′‐Tetramethylbenzidine (TMB), for colormetric analysis that is limited by low sensitivity of absorbance readings and reduced dynamic range, especially at the upper limit of detection. Additionally TMB may be cytotoxic. We have developed a set of novel green and red emitting fluorescent HRP substrates to address these limitations. The fluorescence detection shows a larger dynamic range detecting IgG concentrations from 0.1 to 1000ng/ml in comparison with TMB which saturates at 100ng/ml. In addition, the signal to noise ratio is improved with up to 11.48x higher signal over background compared to just 2.67x for TMB. The substrates also show low well to well variation with standard deviation of <3%. Since benzidine compounds are known to exhibit health concerns, cytotoxicity assays were performed in HeLa cells. After 3 days incubation with the substrates using the MTT assay, these new substrates demonstrated significantly less toxicity then TMB. In conclusion we present a set of new substrates for HRP‐ELISA that have desirable characteristics for use in IgG quantification.
STRUCTURAL ANALOGS OF PULMONARY SURFACTANT PHOSPHATIDYLGLYCEROL ANTAGONIZE ACTIVATION OF TOLL‐LIKE RECEPTORS 2 AND 4
Authors: Kandasamy Pitchaimani , Berry Karin A Zemski , Leslie Christina C , Murphy Robert C , Voelker Dennis R
Published: June 2021

The pulmonary surfactant phospholipid, palmitoyloleoylphosphatidylglycerol (POPG), inhibits TLR2 and TLR4 signaling from the cell surface of macrophages. In this study we sought to develop structural analogs of POPG that vary in polar head group length, hydroxylation and alkyl branching, as novel compounds for suppressing the TLR inflammatory responses. These analogs were synthesized using a transphosphatidylation reaction. Analogs of POPG with C3 and C4 alkyl head group length (POP‐PROPANOL and POP‐BUTANOL) are less effective than POPG as TLR2 and TLR4 antagonists. However, adding a hydroxyl group at the alkyl chain 2, 3 or 4‐position (POP‐PROPANEDIOLS or POP‐BUTANEDIOLS) restored their inhibitory effects against TLR2 and TLR4 agonists. Interestingly, analogs with C1 and C2 alkyl head group length (POP‐METHANOL and POP‐ETHANOL) were effective as LPS antagonists. POP‐DIMETHYLPROPANEDIOL contains 2 methyl groups at the head group alkyl 2 position; and this analog is effective for inhibition of an LPS stimulus. However, it is a weak inhibitor of MALP‐2 induced AA release. Addition of amino group at the alkyl 2 position (POP‐SERINOL) completely abolished the potency of analogs. Collectively, these findings identify new compounds for antagonizing TLR2 and TLR4 activation and define structural properties for discriminating between the two receptor systems. NIH‐HL094629, COLORADO C2D2.
Defects in galactose metabolism and glycoconjugate biosynthesis in UDP‐glucose pyrophosphorylase‐deficient fibroblasts are reversed by supplementing the cell growth medium with galactose
Authors: Edward Stuart , Moore Hanson , Durrant Christelle , Willemetz Alexandra , Sala Giusy , Chrétien Dominique , Ghidoni Riccardo , Rötig Agnès , Thelestam Monica , Katz Abram , Ermonval Myriam
Published: June 2021

UDP‐glucose (UDP‐Glc) is required for galactose metabolism and glycoconjugate biosynthesis and is synthesised from glucose‐1‐ phosphate by UDP‐Glc pyrophosphorylase (UGPP). UGPP‐deficient fibroblasts displaying reduced UDP‐Glc, impaired mitochondrial function and increased glucose regulated protein expression have been described. We have now characterised glycoconjugate biosynthesis and galactose metabolism in these cells and show that they display: 1) reduced [6‐3H]galactose incorporation into UDP‐[6‐3H]‐galactose; 2) an inability to glucosylate the dolichol‐linked oligosaccharide required for protein N‐glycosylation; 3) hypo‐N‐glycosylated PrPC; 4) truncated O‐glycans, and 5) reduced glucosylceramide and NeuAcα2,3Galβ1,4Glcβ‐ceramide levels. Transfection of the cells with wild type UGPP restores all the above defects except mitochondrial function. Surprisingly, it was discovered that the effects of transfecting the mutant cells with wild type UGPP were reproduced by prolonged cultivation of the cells with galactose. Accordingly, we describe a novel reversable model with which to study defects in galactose metabolism and glycoconjugate biosynthesis.
Alterations in the metabolomics of S‐containing substances in rats after betaine intake
Authors: Ahn Chul Won , Kwon Do Young , Kim Sun Ju , Jun Doo Sung , Lee Yoo Min , Kim Young Chul
Published: June 2021

Previous works have shown that betaine administration results in significant changes in hepatic metabolism of S‐amino acids. In this study we examined the changes in the metabolomics of S‐containing substances in kidney of the rats fed betaine for 2 wk. Renal methionine level was increased by 42 % in the betaine‐fed rats. However, betaine‐homocysteine methyltransferase (BHMT) protein or its activity in kidney was not altered. Methionine synthase that mediates the other remethylation reaction of homocysteine, was also unaffected. Betaine intake resulted in an elevation of S‐adenosylmethionine and induction of methionine adenosyltransferase II. But the level of S‐adenosylhomocysteine, total homocysteine, cysteine, or cystathionine β‐synthase protein was not changed in kidney. Betaine intake did not affect the renal glutathione level or expression of glutamylcysteine synthetase that mediates its synthesis. On the other hand, cysteine dioxygenase was reduced markedly in the rats fed betaine. Cysteine sulfinate decarboxylase expression and hypotaurine were increased by betaine intake. These results indicate that dietary betaine affects the renal S‐amino acid metabolism significantly despite the deficiency of BHMT in kidney. The elevation of methionine concentration in kidney may be accounted for by an increase in this amino acid in liver.
Effect of lycopene supplementation on glucose and lipid metabolism in high fat diet‐induced obese Mongolian gerbils
Authors: Choi Soo-Kyong , Seo Jung-Sook
Published: June 2021

Lycopene, a carotenoid present predominantly in tomatoes, is one of the powerful antioxidant compounds. The purpose of the present study was to investigate the effect of lycopene supplementation on glucose and lipid metabolism in obese gerbils. Gerbils were fed on each experimental diet for 6 weeks, a normal diet (NC), a normal diet with lycopene (NL), a high fat diet (HF), a high fat diet with lycopene (HFL). Plasma glucose and insulin levels decreased in the HFL groups compared with the HF group. Lycopene feeding led to decrease the hepatic 3‐hydroxy‐3‐methylglutaryl coenzyme A reductase and acyl‐coenzyme A : cholesterol acyl‐transferase activities in gerbils. The hepatic fatty acid synthase activity was higher in the HF group than the NC group, but it was reduced by lycopene supplementation compared with that of control group. Significant increases in carnitine palmitoyltransferase and ¥â‐oxidation activities were observed in the groups supplemented with lycopene compared with those in the control group. Lycopene supplementation significantly elevated hepatic malic enzyme and glucokinase activities. These results suggest that lycopene supplementation would be efficient for the prevention of obesity‐related metabolic diseases. This research was supported by Basic Science Research Program through the National Research Foundation of Korea funded by the Ministry of Education, Science and Technology.
Tetranectin Promotes Adipogenesis and Lipogenesis and is Negatively Regulated by Polyunsaturated Fatty Acids in Human Adipocytes
Authors: Wang Ya-Chin , Ding Shih-Torng
Published: June 2021

Adipose tissues secrets factors, or adipokines, regulate body energy homeostasis. To search for novel adipokines, we employed 2‐D protein gel electropherosis and LC‐MS/MS to analyze human primary adipocyte culture medium, in which tetranectin (TN). We found that the mRNA expression of TN is positively correlated with the differentiation and lipid accumulation of human preadipocytes, as well as the body mass index. Overexpression and knockdown of TN in human primary adipocytes by lentivirus prove that TN increases the expression of adipogenic and lipogenic genes, such as peroxisome proliferator‐activated receptor gamma, lipoprotein lipase and fatty acid synthesis, but decreases those related to lipolysis, including hormone sensitive lipase, interleukin 6 and serum amyloid A along with altered profiles of proteins responsible primarily for lipid metabolism, as analyzed by quantitative proteomic analysis and quantitative real‐time PCR. In addition, treatments of polyunsaturated fatty acids (PUFA) suppressed TN expression. Taken together, our data suggest that not only TN potentially serves as a novel adipokine contributing to the regulation of energy homeostasis and lipid metabolism, but also components of the TN‐associated signaling pathways the therapeutic targets for treating obesity and associated metabolic diseases.
Dietary Intake of Added Sugars Is Associated with Diastolic but not Systolic Blood Pressure in Children
Authors: Kell Kenneth P , Fernandez Jose R
Published: June 2021

The prevalence of pediatric elevated blood pressure (BP) in the US has been steadily increasing. The etiology for this phenomenon is not fully understood; however, concerns that elevated BP during childhood may translate into adult hypertension merit the evaluation of lifestyle factors that could help preserve pediatric cardiovascular health. This study explored the relationship between added sugars (sugars not naturally occurring in foods) in the diet and pediatric BP using a sample of 310 children aged 7–12 of diverse backgrounds. Diastolic and systolic BP measures were obtained by averaging eight measurements taken on two consecutive days during an in‐clinic visit. Two 24‐hour dietary recalls were conducted to assess dietary intakes. A multivariate regression was performed to evaluate the relationships between added sugars and BP, sodium and BP, and both sodium and added sugars with BP. All models were controlled for age, sex, total calories, ethnicity, physical activity, and body mass index. The relationship between added sugars and diastolic BP was significant and positive, with and without sodium in the model (p < 0.05). Sodium was marginally associated with diastolic BP (P = 0.0720). No significant association was seen between added sugars and/or sodium and systolic BP. These results suggest a physiological connection between added sugars and pediatric elevated diastolic BP that may be prevented via diet.
NK cell populations and cytotoxic activity are greater in pigs fed mother's milk than formula
Authors: Liu Kilia , Comstock Sarah S. , Burdette Jill M. , Monaco Marcia H. , Donovan Sharon M.
Published: June 2021

Natural killer cells (NKC) provide an immediate response to fight against infections. Herein, how diet influences NKC numbers and cytotoxic activity was investigated. Using flow cytometry, NKC (CD3‐CD4‐CD8+) populations were identified in the peripheral blood mononuclear cell (PBMC), mesenteric lymph nodes (MLN) and spleen of 21 day‐old sow‐reared (SR, n=9) and formula‐fed (FF, n=8) pigs. For cytotoxicity assays, NKC were negatively selected from PBMC and incubated (2h) with 20ng/ml IL‐2 followed by an incubation (48h) with DiOC18 labeled K562 target cells at a 10:1 effector‐to‐target ratio. Propidium iodide was added to identify killed cells. Cytotoxicity is reported as killed target (PI+DiOC18+) cells as a percent of total target (DiOC18+) cells using spontaneous target cell death as a normalizer. NKC population were greater in PBMC, MLN, and spleen of SR than FF (p<0.05). IL‐2 increased the cytotoxicity of NKC from both SR and FF, but NKC from blood of SR exhibited higher cytotoxicity than those from FF (p<0.001). In summary, NKC activity was greater in SR than FF piglets, providing a potential mechanism whereby mother's milk improves neonatal innate immune response and defense against microbial infections.
Can a high fiber diet improve bowel function and health‐related quality of life in patients with Crohn's disease?
Authors: Brotherton Carol S. , Taylor Ann Gill , Anderson Joel G.
Published: June 2021

The objective of this pilot study was to test feasibility and efficacy of a dietary instruction featuring wheat bran cereal consumption and refined carbohydrate restriction in a sample of individuals with active symptoms of Crohn's disease. A 28‐day randomized, controlled, single‐blind design was used to measure health‐related quality of life and gastrointestinal function over time in the intervention group (n = 4) and the attention control group (n = 3). Results demonstrated that consuming one‐half cup of wheat bran once a day was feasible and caused no adverse effects. Participants consuming wheat bran reported improved health‐related quality of life (p = 0.028) and gastrointestinal function (p = 0.008) compared to the attention control group that received general dietary instructions. A 44‐point increase on the Inflammatory Bowel Disease Questionnaire demonstrated a clinically significant change. This study supports the idea that, in the absence of contraindications, a dietary pattern featuring daily consumption of concentrated wheat bran cereal and refined carbohydrate restriction can be easily adopted by individuals with moderately active symptoms of Crohn's disease and may improve health‐related quality of life and decrease or eliminate gastrointestinal symptoms. This study was supported by grant number F31‐NRO11121 from the National Institute of Nursing Research, National Institutes of Health.
Grapefruit consumption is associated with benefits to the intake of certain nutrients, body composition, and select physiologic parameters in U.S. women
Authors: Murphy Mary M , Barraj Leila M , Bi Xiaoyu , Rampersaud Gail C
Published: June 2021

The purpose of this study was to estimate usual intake (UI) of total grapefruit (GF) (fruit and juice) by women age 19+ y and to compare certain nutrient intakes and select health‐related parameters between consumers (GFC) and non‐consumers (GFNC) using data collected in NHANES 2003–8. GFC (n=180) were defined as women reporting intake of GF on either of 2 days of dietary recall. UI of vitamin C by GFC was significantly higher (p<0.0001) than intake by GFNC (n=6219) (145±3.9 vs 76±1.2 mg/d); UI of beta‐carotene, vitamin B6, magnesium, fiber and potassium by GFC also were higher compared to GFNC while intakes of added sugars, total fat, saturated fat and cholesterol were lower (p<0.05); energy intakes were comparable. Mean body weight (70.2±1.6 vs 75.4±0.5 kg), waist circumference (91±1.5 vs 95±0.4 cm) and BMI (26.9±0.6 vs 28.6±0.2 kg/m2) of GFC were significantly lower (p<0.01) as compared to GFNC, though the risk of overweight/obesity did not differ. HDL‐cholesterol levels were significantly higher (p<0.01) in GFC (63±1.8 vs 58±0.3 mg/dL) and triglyceride (TG) levels were lower (106±5.7 vs 130±2.5 mg/dL). There were no differences between groups in serum total or LDL‐cholesterol, blood pressure, plasma glucose or insulin. GF consumption may have a positive impact on nutrient intakes and is associated with lower body measures and higher HDL‐cholesterol among women. Supported by the Florida Department of Citrus.
The study of dessert preferences of Korean local food restaurant customers
Authors: Khil Jinmo , Kim Song-Suk
Published: June 2021

This study was conducted to investigate the type and the preference of dessert served after Korean local Foods by local foods restaurant customer in GyeongBuk area. The subjects were 373 customers consisted of 108(29.0%) men and 265(71.0%) women. The subjects answered that it is important to develop Korean traditional dessert instead of instant coffee which was usually served. The results showed that Korean traditional beverages such as ‘sikhye’ and ‘sujeonggua’ were mostly preferred by 45.4% of man, 47.2% of woman who answered the questionnaire. Next preferred dessert menu was traditional Korean hot tea(Han cha) by 33.3% of man and 39.6% of woman replied. The preference about coffee was 12% by male and 6.4% by female. Some customers selected Korean rice cakes and cookies rather than beverages. The rice cake was preferred by 4.6% of men and 3.8% of women. The order of dessert preferences were beverages‐hot tea‐coffee‐rice cake and cookies. This study suggested that the local foods restaurant owner and chef should consider developing new dessert item and serving traditional Korean dessert after local foods to satisfy customers.
Body mass index was positively correlated with blood triglyceride and total cholesterol levels
Authors: Kim Young-Nam , Kim Sungah , Cho Youn-Ok
Published: June 2021

To investigate the associations among anthropometric and selected biochemical indices, fasting blood samples were obtained from 208 healthy adults (62 males, 146 females), 20–64 y old, living in South Korea. Weight and height were measured, and body mass index (BMI) was calculated. According to WHO BMI cut‐offs for Asians, 5.8% were underweight; 40.9%, normal weight; 28.9%, overweight; and 24.5%, obesity. Plasma triglyceride levels (TG) of 43.6% were >=150 mg/dl and 22.1% had plasma total cholesterol levels (TC) >=200 mg/dl. TG of overweight and obese subjects were significantly higher than those of underweight and normal subjects (p<0.05). TC of obese subjects was significantly higher than those of underweight and normal subjects. Total protein and albumin levels were positively correlated with height (r=0.32237, p<0.0001; r=0.16231, p=0.0192, respectively). TG and TC had positive correlations with BMI (r=0.24891, p=0.0003; r=0.20752, p=0.0026, respectively), but HDL‐cholesterol levels had a negative correlation with BMI (r=−0.24689, p=0.0003). The findings of this study indicate that BMI is positively correlated with blood lipid concentrations, TG and TC. To reduce a risk of cardiovascular diseases related to hyperlipidemia, weight control programs with nutritional education may be of benefit to Koreans. Supported by 2011 research fund of National Research Foundation of Korea (2011‐0021273).
In vitro comparison of antioxidant capacity of cumin (Cuminum cyminum L.) oils extracted and purified by supercritical fluid extraction and molecular distillation and their main components
Authors: Chen Qinqin , Gan Zhilin , Yang Yang , Li Shuyan , Li Jingming , Ni Yuanying
Published: June 2021

The antioxidant activities (AOAs) of β‐pinene, p‐cymene, γ‐terpinene, cuminaldehyde, cumin oleoresin (COR, supercritical fluid extraction) and its molecular distillation products (CEO, DR&CT) were investigated in present study. IC50 value and kinetics rate were used to evaluate the efficiency of tested samples in scavenging the alkylperoxyl radical (AR) generated in the β‐carotene‐linoleic acid emulsion system and the 2,2‐diphenyl‐1‐picrylhydrazyl radical (DPPH·). In the kinetic approach, antioxidant efficiency was estimated at t=20 s and expressed as the rate of β‐carotene remaining (R1(β‐carotene)) and the rate of DPPH· scavenging (R2(DPPH)). Results showed that the order of AOAs of cumin oils was DR=COR >CEO >CT. Among all samples, γ‐terpinene exhibited the highest R1(β‐carotene) value and the lowest IC50 value (0.73mg/ml), which indicated the strongest response in AR scavenging in β‐carotene bleaching test. The R2(DPPH) values for DR and COR, followed by γ‐terpinene, showed the highest response in scavenging DPPH·, with IC50 values were 6.72, 8.53, 22.73mg/ml, respectively. There were good correlations between IC50 DPPH and IC50 β‐carotene (r2=0.7990), but no association could be found between R1(β‐carotene) and R2(DPPH). Our results showed that one of the active antioxidant components in cumin oils was γ–terpinene.
MTHFR 1298 A>C gene polymorphism is associated with Non‐Alcoholic Fatty Liver Disease (NAFLD) and Insulin Resistance in well‐nourished patients
Authors: Trovato Guglielmo M , Ragusa Angela , Catalano Daniela , Martines G. Fabio , Tonzuso Antonia , Pirri Clara , Buccheri Maria Antonietta , Trovato Francesca M , Di Nora Concetta
Published: June 2021

MTHFR gene encodes MethyleneTetraHydroFolate Reductase, a regulatory enzyme whose polymorphisms are associated with higher homocysteine. Among polymorphisms, C677T, a termolabile form, but not A1298C, was associated with fatty liver and insulin resistance.

Aim

to investigate the influence of A1298C polymorphism on NAFLD in well‐nourished non‐diabetic patients (pts).

Patients

263 subjects (121 M, 142 F), age 53,19±16,33 years, BMI 28,08±5,92 Kg/m2, on adequate diet. A1298C and C677T polymorphisms were identified by RFLP‐PCR in peripheral blood; 100 C677T polymorphism and 69 double heterozygous pts were excluded by the analysis. Insulin resistance was assessed by HOMA; NAFLD by UltraSound (BLS). Finally 94 subject are included: 26 MTHFR 1298 AA, 35 MTHFR 1298 CC and 33 MTHFR 1298 AC.

Results

The three groups were not different for age, gender, dietary profile and BMI. Control group (MTHFR 1298 AA, no polymorphism) vs. MTHFR 1298 AC (eterozygotic) pts had more severe NAFLD (BLS: 0,54±0,76 vs. 1,12±1,14; p<0.029), insulin resistance (HOMA 2,12±1,12 vs. 3,20±2,35; p<0.036) and higher ALT/GGT. The MTHFR 1298 CC (omozygotic) pts did not show any difference from MTHFR 1298 AA.

Conclusion

MTHFR 1298 A>C gene polymorphism confers an increased risk for NAFLD and Insulin resistance, diet‐related conditions, with same level of obesity. Nutritional and omozygotic relationship deserve further investigation.
Lack of thrombospondin‐1 increases tumorigenesis and decreases survival of in a new mouse model of colorectal cancer
Authors: Emenaker Nancy J. , Soto Pantoja David R. , Roberts David D.
Published: June 2021

According to the National Cancer Institute more than 140,000 people will be diagnosed in the United States with colorectal cancer this year alone. The adenomatus polyposis coli (APC) gene is frequently mutated in colorectal cancer, the APC‐min mouse correspondingly is a widely used model for the study of this type of cancer. Yet, intestinal tumors develop predominately in the small intestine reflecting familial adenomatous polyposis rather than colorectal cancer. Studies indicate that the matricellular protein Thrombospondin 1 (TSP1), a key component in modifying inflammatory processes, is lost during the development of colon cancer. To study the role of TSP1 during inflammation driven colorectal carcinogenesis we developed an APC‐min and TSP−/− APC‐min mouse model. A lack of TSP1 increases tumor size and multiplicity predominately in the large intestine of these mice. Our data indicate that expression of TSP1 in the APC‐min mouse prevents the development of adenomas in the large intestine in mice fed a low fat diet. Moreover, TSP−/− APC‐min mice show 50% reduction in survival when compared to TSP+/+APC‐min mice alone (P<0.05). Our in vitro experiments indicate that this difference may be due to TSP1 regulation of pro‐inflammatory mechanisms. Together this data indicates TSP1 regulates progression of colon carcinogenesis and define cellular and molecular targets of TSP1 in the prevention of colorectal cancer.
Screening for antihypertensive activities of traditional herbs in Donguibogam
Authors: Ok Hyang Mok , Jung Saet Byeol , Choi Ye Jin , Kim Do ram , Kim Ji Yeon , Kwon Oran
Published: June 2021

The aim of this study is to examine the all the current science‐based evidence supporting the use of Korean medicinal plants. It was designed to search traditional Korean herbal remedies with specific functionality, antihypertension, in old literatures and systemically prove their effectiveness from a comprehensive, upto‐date literature review and in vitro tests. For a terminology matching related to hypertension symptom, we used primarily Korean Classification of Disease in Oriental Medicine (KCDOM) and Korea Traditional Knowledge Portal (KTKP) databases. Water or ethanol extracts of all the screened plants used in this study were purchased from the Korea Research Institute of Bioscience and Biotechnology (KRIBB). Herbal preparations were investigated in vitro on influence on nitric oxide production and ACE inhibition activity. We used the 15T FT‐ICR MS to identify the chemical fingerprint of major active compounds contained in the selected plants. We found out seventeen kinds of symptoms relevant to hypertension using KCDOM and KTKP and seventy one plants extracts in Donguibogam. The six most potent herbs were identified as Sophorae fructus, Angelicae koreanae, Melonis calyx, Evodiae fructus, Bambusae caulis, and Nepetae spica through in vitro analysis. This study suggests the potential use of traditional plants for antihypertension.
Effect of polyphenol rich dark chocolate ingestion in SOD2 and IL‐6 expression and plasma total antioxidant capacity
Authors: Verástegui-Tena Luz Maria , Hernández Brianda A , Tolentino Maricruz , Barrera Paloma K , Perez-Lizaur Ana Bertha , Tejero Maria E
Published: June 2021

Background

Recent studies have demonstrated that ingestion of polyphenol rich foods can increase plasma antioxidant status and cause a decrease of inflammatory markers. The aim of this study is to evaluate the effect of the ingestion of a polyphenol rich dark chocolate in plasma total antioxidant capacity and in the expression of SOD2 and IL‐6.

Methods

Eight healthy volunteers (age 25.75±6.036, BMI 24.9±4.238) were part of a crossover pilot study. Participants were non‐smokers and taking no antioxidant supplements. All volunteers ingested 40 g of a polyphenol rich dark chocolate (70% cacao) and 40 g of a control chocolate (32% cacao). Blood samples were taken at baseline and 2 hours after chocolate ingestion. Total plasma antioxidant capacity was evaluated using the TBARS assay. Expression of mRNA levels of IL‐6 and SOD2 was evaluated in whole blood using Taqman RT‐PCR.

Results

A significant increase in SOD2 expression was observed after dark chocolate ingestion (p=0.031), but not after control intake. No significant difference was found in IL‐6 expression and TBARS levels before and after consumption of each chocolate, or between treatments.

Conclusions

Dark chocolate ingestion may increase total antioxidant capacity by inducing SOD2 expression. Further studies are required to identify the mechanisms underlying chocolate polyphenols activity.

Source of research support Nestlé México.
High‐fat diet‐dependent increase in plasma immunoglobulin levels are repressed by aldehyde dehydrogenase1 a1
Authors: Meyers Joseph Michael , Fluegge Keith , Yasmeen Rumana , DiSilvestro David , Yang Fangping , Ziouzenkova Ouliana , Duester Gregg
Published: June 2021

Abnormal immunoglobulin (Ig) production underlines hematological cancers. Studies show association of these cancers with obesity. However, it is unclear how high‐fat (HF) diets influence Ig levels in the circulation. We used six 59 weeks‐old C57BL/6 mice (wild type WT), fed a diet of 5.67% by calorie of saturated fat, compared to six age and sex‐matched WT mice fed a HF diet of 45% by calorie saturated fat for 42 weeks. Using ELISA we quantified Ig classes (IgG1, IgG2a, IgG2b, IgG3, IgA, IgM, Igκ and Igλ) in the plasma of both groups. The intra and inter‐assay variations of the ELISA assay were 2–12% and 5–30% respectively. Mice fed regular and HF diets have similar proportions of Ig's with exception of a 9% increase in IgLκ in HF diets. There was a quantitative increase of 200% to over 500% in Ig's of every class present in plasma from mice on HF diets. However, the increase in Ig production on a HF diet was suppressed by retinoic acid‐generating enzyme aldehyde dehydrogenase 1a1 (Aldh1a1). We studied this relationship in Aldh1a1−/− mice (n=6) vs. WT (n=6) mice on HF diets. Aldh1a1 −/− mice exhibited an increase in plasma Ig's: IgG1 224%, IgG2a 978%, IgG2b 106%, IgG3 217%, IgA 156%, IgLλ 105%. We confirmed polyclonal gammopathy in Aldh1a1−/− compared to WT mice on HF diet using serum protein electrophoresis. We conclude that dietary fat content increases Ig concentration in plasma dependent on the expression of Aldh1a1 enzyme.
Collagenase‐3 derived peptide inhibitor is chondro‐protective in a rat model of osteoarthritis
Authors: Holmes Tiffany C , Lau Shukkwan K , Youngwirth Jonathan , Klunk James , Lombard Frederick J , Earland Noah , Pinkney Nathaniel , Palumbo Dante , Rutkowski Lorrine , Denzine Mark , Kubinski Andrew , Maugle Tyson , Lukashova Lyudamila , Teixeira Cristina , Selim Abdulhafez A , D'Angelo Marina
Published: June 2021

Osteoarthritis (OA) is a disease of the joints caused by an imbalance between extracellular matrix destruction and production. Some of the molecular components responsible for this imbalance are matrix metalloproteinases (MMPs), especially, collagenase‐3 (MMP13) which degrades collagen type II. We have tested the efficacy of an MMP13‐derived peptide inhibitor (PxTx001‐1) in altering the pathology of an established rat model of OA. Utilizing the peptide as an early intervention therapeutic, we have assessed the ability of our inhibitor to affect the progress of OA. Peptide or control saline was injected into the OA knee joint for four consecutive weeks. Endpoint assessment of: toxicity, measured by CBC and serum chemistry; joint space narrowing, measured by X‐ray; joint functionality, measured by stride test; and tissue pathology, measured by micro computed tomography and histology were completed. All data indicates that the inhibitor is non‐toxic and diminishes the pathology associated with OA. We are currently testing this inhibitor's disease modifying properties in a canine model of OA. This work was supported, in part, by intramural funding.
Interleukin‐17A promotes inflammation and hepatocellular injury during obstructive cholestasis
Authors: OˈBrien Kate M , Copple Bryan L
Published: June 2021

In patients with cholestatic liver disease, serum bile acid concentrations are increased resulting in hepatic inflammation, injury, and fibrosis. The molecular mechanism by which cholestasis stimulates liver inflammation is not known. Recent studies demonstrated that the inflammatory cytokine, interleukin‐17A (IL‐17A), is increased in these patients. Whether IL‐17A contributes to the pathogenesis of this disease, however, is not known. Therefore, the hypothesis was tested that IL‐17A is required for liver inflammation and injury during cholestasis. In mice subjected to bile duct ligation (BDL), a model of obstructive cholestasis, IL‐17A mRNA levels were increased in the liver. Inhibition of IL‐17A in BDL mice with a neutralizing antibody reduced serum alanine aminotransferase levels and areas of hepatic necrosis when compared to BDL mice treated with control IgG. Neutralization of IL‐17A also partially prevented upregulation of several proinflammatory chemokines, including macrophage inflammatory protein‐2, Ccl7, and Cxcl5. We previously demonstrated that bile acids at pathological concentrations stimulate hepatocytes to produce chemokines. Interestingly, pretreatment of hepatocytes with IL‐17A synergistically enhanced production of MIP‐2 by hepatocytes. These data suggest that IL‐17A and bile acids interact to stimulate inflammation and liver injury during obstructive cholestasis. This work was supported by the National Institutes of Health National Institute of Diabetes and Digestive and Kidney Diseases [Grant DK073566].
Effects of antenatal steroid and tracheal occlusion on pulmonary development in the fetal rabbit model of congenital diaphragmatic hernia
Authors: DEMIRBAG Suzi , Ilhami Surer , Yesildaglar Narter , Onguru Onder
Published: June 2021

Objective

Congenital diaphragmatic hernia is one of the life threatening malformations that occurs 1 in 2,400 births. The aim of the study was to investigate the effects of fetal tracheal occlusion (TO) and steroids on pulmonary development in rabbits with surgically‐created diaphragmatic hernia.

Methods

There were four groups in the study: Sham (n=10), tracheal occlusion (n=5), dexamethasone (n=5) and control (n=5) groups. On day 23 of gestation (pseudoglandular stage) CDH was created in 15 fetuses of five time‐mated pregnant New Zealand type white rabbits. TO and fetal intraperitoneal dexamethasone administration (0.25 mg/kg) were performed on day 28. Fetuses were harvested by cesarean section on day 30 of gestation. Pulmonary development was evaluated by lung‐to‐body weight ratio (LBWR).

Results

Congenital diaphragmatic hernia decreased the LBWR in control group. TO and dexamethasone increased the LBWR. There were significant differences between control and TO (0.0289±0.0037vs 0.0348±0.0048, p<0.05), and control and dexamethasone groups (0.0289±0.0037 vs. 0.0301±0.0033, p<0.05) for LBWR.

Conclusion

Both tracheal occlusion and fetal intraperitoneal dexamethasone administration can correct pulmonary development in rabbits with surgically created CDH.
Nuclear VCP acts as a tumor promoting factor for oral squamous cell carcinoma
Authors: Sheu Jim Jinn-Chyuan , Li Chien-Feng , Lai Chien-Chen , Chiu I-Wen , Chen Chih-Mei
Published: June 2021

The objective of this study is to characterize the oncogenic role of a recently identified cancer‐associated gene called VCP (also known as p97 or CDC48) in oral cancer and its clinical implications for developing new molecular diagnostics and therapy. Based on a genomewide study, this gene was found located at the 9p13 amplicon in oral squamous cell carcinomas (OSCC). In addition, VCP gene has been reported highly upregulated in a variety of tumor samples. Thus, it is believed that VCP gene plays a crucial role in cancer development and progression. Our preliminary data indicate that VCP amplification correlates with nuclear localization of its protein and poor clinical outcome, suggesting VCP partners with several nuclear proteins to form a functional complex that may provide growth advantages for oncogenic signaling and cancer aggressiveness. Blocking the assembly of nuclear VCP complex could be a potential therapeutic approach to treat oral cancer patients with VCP amplification.
A subgroup of lymphangioleiomyomatosis with no TSC mutations identified
Authors: BADRI KAMESWARA RAO , Qin Wei , Gao Ling , Zhe Xiaoning , Gong Can , Schuger Noa , Hyjek Elizabeth , Kwiatkowski David J , Schuger Lucia
Published: June 2021

Lymphangioleiomyomatosis (LAM) is a rare disease characterized by infiltration of the lungs by neoplastic cells with smooth muscle and melanocytic differentiation. Dissemination of LAM cells cause progressive cystic destruction of the lungs, leading to respiratory insufficiency and death. LAM occurs either sporadically (S‐LAM) or in association with tuberous sclerosis (TSC). TSC is caused by mutations in the genes coding for tuberin (TSC2) and hamartin (TSC1), which dimerize and inhibit the mTOR pathway of cell growth and proliferation. LAM was thus thought to follow a two‐hit model leading to complete inactivation of TSC2 (or TSC1) in LAM cells. In this study, we sequenced TSC2 and TSC1 genes in lung samples from 10 S‐LAM patients. Laser capture microdissection was used to isolate LAM cell clusters, and deep sequencing was performed to detect mutations with greater sensitivity than was previously possible. We identified TSC2 mutations in 8 out of the 10 cases of S‐LAM. Four of these cases bore 2‐hit TSC2 inactivating mutations. Each of the other 4 cases, however, displayed a single mutation which was restricted to a small fraction of LAM cells. More importantly, no TSC2 or TSC1 mutations were identified in 2 of our 10 S‐LAM cases. Our findings are consistent with the existence of a form of S‐LAM unassociated with mutations in TSC2 or TSC1 and that of LAM cells lacking TSC2 mutations.

Funded by NHLBI. KB is ALA Dalsemer Scholar
Synergistic effect of Biochanin A on antiproliferative potential of Atorvastatin
Authors: Tadinada Satya Murthy , Lai James C.K. , Bhushan Alok
Published: June 2021

Pancreatic cancer, with its dismally low survival rate, is one of the most aggressive forms of cancer and is the fourth leading cause of cancer‐related deaths in the United States. Gemcitabine, the first line drug approved by the FDA has not been a huge therapeutic success because it improves the survival by just six months. Epidemiological studies implicated a role for dietary isoflavones in lower incidence of different cancers in the Asian population. Isoflavones such as genistein have been tested for their anti‐cancer potentials in different cancers. Primarily used in management of hyperlipidemia and cardiovascular diseases, statins have been documented to exert anti‐proliferative effects. We have therefore hypothesized a combination treatment with an isoflavone (i.e., biochanin A) and a statin (i.e., atorvastatin) exerts synergistic anti‐cancer effects on pancreatic cancer cells. Our ongoing studies have demonstrated that biochanin A and atorvastatin synergistically lowered survival of AsPC1 and MIAPaCa‐2 pancreatic cancer cells. Concomitant with lowering survival of these cancer cells, biochanin A and atorvastatin combination treatment also induced morphological changes (e.g., shape & cell rounding) in these cells. Our ongoing studies to elucidate signaling mechanisms underlying the combination treatment effects suggested that the PI3K/Akt pathway was unlikely to be implicated in the combination treatment‐induced death of pancreatic cancer cells. Thus, our findings may have therapeutic potential in the design of new combination therapy for pancreatic cancer.

Grant support‐ INBRE Program, NIH Grant No. P20 RR016454, Mountain States Tumor and Medical Research Institute and University Research Committee, ISU grant.
Effects of GPER activation on intracellular Ca2+ stores in the vascular endothelium
Authors: Burgard Michelle , Giles Jennifer , STENCEL MICHAEL , TRAN QUANG-KIM
Published: June 2021

The novel G protein‐coupled estrogen receptor 1 (GPER) has been demonstrated to mediate numerous beneficial cardiovascular effects. We have begun to investigate the effects of GPER activation on intracellular Ca2+ stores in the vascular endothelium. We observed that GPER is present in primary endothelial cells in a glycosylated form. Immunostaining locates GPER at both the plasma membrane and in the perinuclear region. In nominally Ca2+‐free condition, treatment with thapsigargin triggers the release of the ER's Ca2+ content. Subsequent addition of the GPER agonist G1 in Ca2+‐free condition surprisingly triggers a slow and sustained Ca2+ rise up to ~ 400 nM, a value comparable to agonist‐induced Ca2+ entry signals. Inhibition of PI3 kinase with wortmannin decreases G1‐triggered Ca2+ signal in Ca2+ free condition, but does not abolish it, suggesting that GPER activation in endothelial cells triggers the release of both IP3‐dependent and IP3‐independent Ca2+ stores. Pretreatment of endothelial cells with brefeldin A, a disruptor of the trans‐Golgi network, abolishes the slow and prolonged phase of the Ca2+ response to G1 treatment. Our data suggest that activation of GPER in primary vascular endothelial cells triggers the release of two different types of intracellular Ca2+ stores, namely the endoplasmic reticulum and the Golgi. The data also suggest that the Golgi represents a substantial intracellular Ca2+ store in the vascular endothelium.
Biological response of commercial high fiber breakfast cereals consumed in northwestern of Mexico in growing rats
Authors: Falcon Maria
Published: June 2021

The importance of dietary fiber and the contribution of fiber‐rich commercial breakfast cereals to the Mexican diet have long been recognized. The objective of this study was to biologically evaluate three commonly consumed high fiber commercial breakfast cereals selected from the local market, two different brand of whole wheat and wheat bran based, which differed from each other in terms of size and quantity of bran, and one rolled whole oat cereal, using Sprague dawley rats. Biological indices determined were food efficiency ratio (FER), net protein ratio (NPR) and in vivo digestibility. In vivo digestibility of diet based on oat cereal was 83 – 85%, superior to diets containing wheat bran, presenting values from 62 – 64% of apparent nitrogen digestibility, and 66 – 67% of true nitrogen digestibility, which corresponds to up to71% of casein digestibility. NPR of diet based on oat cereal was 3.2 also superior to diets containing wheat bran, presenting values from 1.8 – 2, which corresponds to up to 46% of casein. Results showed that commercial breakfast cereals with different type and level of dietary fiber showed significant differences in protein quality. This study indicates that fiber in wheat bran products reduces nitrogen utilization in growing rats by causing increases in excretion of endogenous and dietary fecal nitrogen. It also shows that some fibers alter the biological value of the nitrogen that is absorbed.
Effect of early diabetes on the expression of alpha‐1 adrenoceptors in aorta of Wistar Kyoto and Spontaneously Hypertensive rats
Authors: Rodriguez Jessica Edith Rodriguez , Albor Aldo Arturo Reséndiz , Martínez Ivonne Maciel Arciniega , Chong Enrique Hong , Rodriguez Rafael Campos , Villafaña Santiago
Published: June 2021

Adrenergic receptors play an important role in the regulation of blood pressure. Diabetes and hypertension affect this mechanism producing changes in vascular responses and exacerbating cardiovascular diseases. Several studies have mentioned that vascular changes are different in the early versus advanced stages of diabetes. For instance, in a recent study by our work group, a decrease in the vasopressor response to noradrenaline was found at 4 weeks of the diabetes evolution in Wistar Kyoto (WKY) and Spontaneously Hypertensive rats (SHR). Among the changes that could take place in the early stages of diabetes to cause this decrease in vascular response is a decrease in the expression of alpha‐1 adrenergic receptors. Therefore, the aim of the present study was to assess the expression of the three subtypes of alpha‐1 adrenergic receptors (alpha‐1A, alpha‐1B, alpha‐1D) in the aorta of WKY and SHR with 4 weeks of the evolution of diabetes. The results showed that whereas the condition of diabetes in WKY rats did not modify significantly the expression of alpha‐1 adrenoceptors, in SHR led to a significative decrease in the expression of both alpha‐1A and alpha‐1B receptors, with no change observed in alpha‐1D expression. In conclusion, the early stages of diabetes produce modifications in the expression of alpha‐1 adrenergic receptors that could explain the decrease found in vascular responses.

This work was supported by a CONACYT grant (#82599) and SIP grant (#20110176) COFAA‐ IPN.
Isolated aortic rings from rats surviving sepsis present enhanced responsiveness to vasopressin and angiotensin II
Authors: de Souza Priscila , Marques Maria Consuelo Andrade , da Silva-Santos José Eduardo
Published: June 2021

Epidemiological studies reveal a high mortality rate among patients that are discharged after an episode of sepsis, when compared to age‐matched people. To investigate if changes in the functionality of cardiovascular system contribute to this high mortality rate, we assessed the responses of aortic rings from 3 groups: control, and sepsis induced by cecal ligation and puncture, 30 (S30), and 60 (S60) days after recovery. The effects of potassium, phenylephrine, angiotensin I, calcium chloride, caffeine, acetylcholine, and sodium nitroprusside were not changed in vessels from sepsis‐surviving rats. However, in S60 group angiotensin II (AII)‐ and vasopressin (AVP)‐induced vasoconstriction were increased by 70 and 78%, respectively, when compared to control. It was accompanied by reduced relaxation in response to Y‐27632, a Rho‐kinase inhibitor, with EC50 of 0.43 (0.33 – 0.55) and 1.03 (0.76 – 1.40) μM in control and S60 groups, respectively. AII and AVP induced augmented contractile responses in aortic rings from S60 rats kept in calcium‐free solution, an event reversed by thapsigargin (an inhibitor of SERCA). Our results suggest that aortic rings from sepsis‐surviving rats display increased reactivity to AII and AVP by mechanisms involving changes in calcium mobilization and sensitization.
Dioscorea batatas stimulates procollagen production in vitro and reduces UVB‐induced MMPs secretion in vivo
Authors: Lee Young Eun , Lee Kyoung-Ryul , Jeon Byoung-Kook , Woo Won-Hong , Mun Yeun-Ja
Published: June 2021

Ultraviolet (UV) B irradiation induces the production of matrix metalloproteinases (MMPs), which are responsible for the degradation or synthesis inhibition of collagenous extracellular matrix in connective tissues, causing skin photoaging. In this study, we examined the efficacy of a yam extract in protecting skin against UVB‐induced damage using cultured human dermal fibroblast neonatal (HDFn) cells and hairless mice skin. Dioscorea batatas (DB) was used the synthesis of procollagen and the release of MMP‐1 in HDFn cells were measured by EIA kit and MMP‐1 assay kit, respectively. UVB radiation was applied to the backs of the mice three times a week for 8 weeks. Mice were randomly divided into four groups, and were topical application with the DB (3% and 6%) or vehicle. DB (50 ug/ml) stimulated procollagen production in HDFn cells by 482.4 ng/ml compared to TNF‐á– treated cell (269.5 ng/ml). Also, pre‐treatment of HDFn cells with DB inhibited TNF‐á–induced release of MMP‐1. In vivo study, we found that DB attenuated UVB‐induced epidermal thickness. DB suppressed the expression of MMP‐3 and MMP‐13 induced by UVB irradiation. Our results show that the DB protects against UV‐induced skin damage in hairless mice. This work was supported by the Ministry of Knowledge Economy, Republic of Korea (70007085).