bluuebunny/crossref_metadata_2025 huggingface dataset:
Standardized Curcuminoids Extract (Curcuma longa L.) Protects against inflammation in Human Umbilical Vein Endothelial Cells (HUVEC)
Authors: Angel‐Morales Gabriela , Noratto Giuliana , Mertens‐Talcott Susanne
Published: June 2021

Vascular inflammation is suggested to play a key‐role in the initiation and progression of atherosclerosis. Turmeric, used as medicinal agent in India, contains polyphenolics known as curcuminoids, including curcumin, demethoxycurcumin, and bisdemethoxycurcumin. This study investigated the potential protective effects of standardized curcuminoid extract (SCE) against vascular inflammation induced by high glucose in human umbilical vein endothelial cells (HUVEC). SCE was characterized by HPLC‐PDA. In this study, high glucose (Glc) (25 mM) increased vascular cell adhesion‐1 (VCAM) interleukin (IL‐8), tumor necrosis factor (TNF) and nuclear factor kappa B (NF‐kB) expression up to 2.1, 1.9, 1.7 and 3.0 fold, respectively, compared to the control and treatment with SCE for 24 h inhibited their mRNA expression by up to 50, 70, 40 and 60%, respectively in a dose dependent manner (0‐25 mg/L), these effects were not due to decreased cell viability as assessed by the MTT test. SCE reversed Glc‐and lypopolysacharide (LPS) induced increase of microRNA 146a, the NF‐kB‐dependent gene up to levels found in control cells (6.25‐25 mg/L).In addition, treatment with SCE for 1h incubation at 25 and 50 mg/L blocked formation of Glc‐induced oxygen species (ROS) by 50 and 70% respectively. The present data suggest that SCE could suppress high glucose induced vascular inflammation generation of ROS in HUVEC.

Grant Funding Source

Texas A&M University
Survey of functional food molecules in prickly pear cactus, bitter gourd, and guava for the dietary management of the Type II diabetes
Authors: Syu Kai‐lo , Tam William , Tam Gregory , Tam Chick
Published: June 2021

Objective

Identify functional food molecules found in prickly pear cactus (Opuntia dillenii), bitter gourd (Momordica charantia), and guava (Psidium guajava) with hypoglycemic properties in the dietary management of Type II diabetes.

Method

Use internet research, peer‐reviewed published papers and books that deal with biochemical components of these fruits and vegetables.

Result

Prickly pear cactus has mucilage, a polysaccharides that showed hypoglycemic properties. Bitter gourd helps regulate blood sugar levels because it contains gurmarin, a polypeptide. It also contains alkaloid momordicine, the bitter component in the gourd. The aqueous extract of guava leaves has a good effect to lower blood glucose. Flavonoid glycosides such as pedunculagin, strictinin, and isostrictinin are the effective constituents that have been used in clinical treatment of diabetes to improve the sensitivity of insulin.

Conclusion

The available data suggest that prickly pear cactus, bitter gourd, and guava have the potential to become a component of the diet or a dietary supplement for diabetic and prediabetic patients. These biochemical components from prickly pear cactus, bitter gourd, and guava open new possibilities of improve and/or prevent the Type II diabetes and/or its clinical complications.
Inhibitory effect of Corni fructus water extract on adipocyte differentiation in 3T3‐L1 cells
Authors: Seo Dong Joo , Chung Mi Ja , Kim Dae Jung , You Jin Kyoun , Choe Myeon
Published: June 2021

Obesity is characterized at the cellular level by an increase in the number and size of adipocytes differentiated from fibroblastic preadipocytes in adipose tissue. Inhibition of adipocyte differentiation is one approach among the anti‐obesity strategies. Corni fructus (SSY) has been used as a folk medicine in Korea. In the present study, we investigated the molecular mechanisms of the anti‐obesity effect of SSY water extract on adipocyte differentiation of 3T3‐L1 cells. The differentiation of 3T3‐L1 preadipocytes cells was induced by hormone cocktail, including dexamethasone, isobutylmethylxanthine (IBMX), and insulin. Effects of SSY treatment on adipocyte differentiation were measured by Oil Red O staining, triglyceride and cell viability assays. In addition, the protein expression of peroxisome proliferation activated receptor‐γ (PPAR‐γ) and AMPK‐activated protein kinase (AMPK) was detected by western blotting. 1000 ppm SSY water extract significantly reduced cell number but decrease in cell number was not due to a cytotoxic effect, because cytotoxicity appeared at greater than 2000 ppm. Triglyceride accumulation and PPAR‐γ expression were decreased by SSY water extract in a dose‐dependent manner but failed to show increase of AMPK in SSY treated cells. These data suggest that the anti‐obesity effect of SSY is mediated by the PPAR‐γ signaling pathways in 3T3‐L1 preadipocytes during differentiation.
Hypoglycemic effect of leucine in diet‐induced obese SD rat
Authors: Kwon Hye Jin , Park Hoon Jung , Kim Mi Kyung
Published: June 2021

The aim of this study was to examine the hypoglycemic effect of leucine (Leu) in 6 month‐old Sprague‐Dawley (n=80) rats. One group of 10 rats was fed control diet (control group) and another group of 70 rats was fed a high‐fat diet for 14 wks (high‐fat group) to induce obesity. Ten rats of each group were killed at this stage and the remaining obese rats were then randomly blocked into 6 groups, according to body weight and raised with experimental diets containing different Leu levels (0g, 0.6g, 1.2g /kg BW) and dietary fat level (4%, 20%, w/w) for 5 wks. The levels of fasting blood glucose and fasting plasma insulin and glucagon were measured. HOMA‐indexes were calculated using fasting blood glucose and plasma insulin concentrations. The level of fasting blood glucose and HOMA‐indexes in 1.2g/kg Leu groups were significantly lower than 0g/kg Leu groups, and the levels of plasma insulin and glucagon in 1.2g/kg Leu groups were tended to be lower than 0g/kg Leu groups. In conclusion, the feeding of Leu might have hypoglycemic effect in diet‐induced obese SD rats.

* This work was supported by Sungshin Cement Co., Ltd. of Seoul, Korea and the second stage of the Brain Korea 21 project in 2008.
Absorption of folic acid and ascorbic acid from beverages
Authors: Monsivais Pablo , Carter Brett , Christiansen Mathew , Drewnowski Adam
Published: June 2021

The purpose of this study was to test for possible differences in the bioavailability of nutrients from three different beverages using a double‐blind, within‐subjects experimental design. The study tested whether the bioavailability of vitamin C (i.e. ascorbic acid or AA) and folic acid (FA) in adult men differed after ingestion of a fortified beverage (Essential VitaminWater, Glacêau®), orange juice (OJ) and 1% milk. Participants were aged 20‐35y, with body mass indexes ranging from 20‐30 kg/m2. Serum concentrations of AA and FA were assayed by serial blood draws, which were made at 30 min intervals for 4.5 hours. Results showed that the concentration profile of AA was significantly greater after ingestion of the fortified beverage compared to OJ. However, the bioavailability of AA in the fortified beverage and OJ was not significantly different, indicating that the higher serum concentration of AA in response to the fortified beverage was due to that beverage's higher AA concentration. In terms of FA, analyses indicated that serum concentration and bioavailability did not significantly differ between the fortified beverage and OJ. Research support provided by Glaceau LLC.
Evaluation of novel soluble fiber dextrins for sugar composition, in vitro digestion characteristics, and glycemic and insulinemic responses using a canine model
Authors: Knapp Brenda Kay , Bauer Laura , Parsons Carl , Swanson Kelly , Fahey George
Published: June 2021

Soluble fiber dextrins (SFD) were evaluated for compositional and in vitro digestion characteristics, and for glycemic and insulinemic responses in dogs. Samples were derived from tapioca starch‐ tapioca‐based SFD1 (T1), tapioca‐based SFD2 (T2), and tapioca‐based SFD hydrogenated (TH), and from corn starch‐ corn‐based SFD1 (C1), corn‐based SFD2 (C2), and corn‐based SFD3 (C3). Tapioca SFD1, T2, and C2 had the lowest free sugar concentration (~2.0%). Intermediate free sugar concentrations were found for C1 and C3 (~23%), and TH resulted in the highest (92%) free sugar concentration. All SFDs had low to intermediate amounts of monosaccharides released after simulated hydrolytic digestion. Corn‐based SFDs resulted in a lower (P<0.05) digestibility than tapioca‐based SFDs (23% and 51%, respectively). Glycemic and insulinemic responses varied widely among SFDs, with an average glycemic and insulinemic response 55% lower than for maltodextrin. Tapioca‐based SFDs had lower relative glycemic responses (RGR) than corn‐based SFDs except for C1 which had the lowest RGR among all SFDs. The insulinemic responses followed the same pattern as the glycemic responses. All SFDs had low free sugar contents with varying degrees of resistance to digestion and attenuated glycemic responses. These ingredients may be suitable candidates for reduced glycemic foodstuffs.
Effect of high saturated‐fat diets on bone mass and endochondral ossification in healthy growing rats
Authors: Macri Vanesa , Juiz Natalia , Gonzales Chaves Macarena , Ponce Graciela , Lanata Eduardo , Rodríguez Patricia , Mandalunis Patricia , Zeni Susana , Friedman Silvia
Published: June 2021

High‐fat diets can produce deleterious effects on bone metabolism in growing animals. Objective: To investigate in healthy growing rats the effect of high saturated‐fat diets on bone mass and endochondral ossification. Weaning male Wistar rats (n=20) were fed ad libitum one of the 2 diets, for 4 wks: control (C; fat=7% w/w) (AIN‐93G) or a high‐fat diet containing 20% w/w of beef‐tallow (BT). Then rats were sacrificed. Total skeleton bone mineral density (BMD) and bone mineral content (BMC) were evaluated by DXA (Lunar DPX‐L).The following histomorphometric parameters were evaluated: Bone volume (BV/TV)(%) and cartilage growth plate thickness (CGP.Th).Statistics: Student's t test & Pearson's correlations coefficients were calculated. Results (as mean±SD): BMD(mg/cm2), C=245±13 vs BT=244±13 (p>0.05); BMC(mg), C=1085±319 vs BT=689± 224; (BV/TV)(%), C=45±5 vs BT=38±3 (p<0.03); total GPC.Th(mm), C=0.44±0.04 vs BT=0.43±0.03 (p>0.05); proliferative zone GPC.Th(mm), C=0.21±0.01 vs BT=0.18± 0.01 (p>0.001).The correlation between BMC vs proliferative zone CGP.Th was ‐0.881 (p<0.05).

Conclusion

High saturated‐fat diets lead to a decreased in BMC associated with a significant increased in CGP.Th as well as a reduction in BV/TV. Based on these findings, we suggest that high saturated‐fat diets acts as a dependent risk factor for bone modeling interfering the acquired of bone mass. Awarded by UBACyT O 008 ‐ O015.
PLASMA ADENOSINE DEAMINASE ACTIVITY IN A GROUP OF CHILDREN SUFFERING CYSTIC FIBROSIS.PRELIMINARY STUDY.
Authors: Perris Paula , Feliu Maria Susana , Slobodianik Nora
Published: June 2021

Mucus of children suffering cystic fibrosis (CF) can avoid the normal absorption of essential nutrients and fat, leading to poor digestion, and increased vulnerability to infections. Preliminary results suggest that nutritional status of children suffering CF was altered. Therefore, some authors have observed increase in the activity of sera Adenosine Deaminase (ADA) in patients with pathologies related with lymphocytes T. In this preliminary paper we analyze if plasma ADA activity on this population in nutritional risk, is altered.

The study included 17 children of both genders with CF (2months‐10 years old); the activity of ADA (U/L) was determined by the method of Giusti and Galanti; data were compared with the age‐matched well nourished control (C ) applying the test of Student. The results expressed as X+ SD were: CF: 34.5±15.6* ;C: 23.0±5.6 (p< 0.001). It was observed an statistical increase in the activity of ADA. These preliminary results suggest depression on the mechanisms of defense that are concomitant with an altered nutritional status. ADA determination might be included in the periodic evaluation of patients with CF. In these patients, the assessment of nutritional status must be included between the main parameters that could influence the evolution of the disease, the prognosis and the survival

Partially supported by University of Buenos Aires (B074)
Regulation of c‐Jun N terminal kinase (JNK) signaling by dual specificity phosphatases (DUSPs) in adipocytes during inflammatory stress
Authors: Ferguson Bradley Scott , Nam Heesun , Hopkins Robin G , Morrison Ron F
Published: June 2021

Chronic inflammation is an important element of pathogenic mechanisms linking obesity to diabetes. Mounting evidence strongly suggests that the JNK signaling pathway plays a central role in mediating the inflammatory process as mice deficient in JNK activity are protected against diet‐induced or genetic models of insulin resistance. While numerous studies have examined the upstream signaling on JNK activation, few have addressed the role of JNK deactivation and its effect on biological outcome. Evidence now indicates a critical role for DUSPs in the deactivation of JNK and other signaling pathways. Using 3T3‐L1 adipocytes stimulated with tumor necrosis factor α (TNFα) as a model of inflammation, we examined the role of DUSPs on JNK deactivation and inflammatory function. We show that the transient activation (<1h) of JNK signaling in response to TNFα is prolonged (>2h) in the presence of transcriptional inhibition with actinomyosin D suggesting a role for inducible phosphatases. Based on these data, we identified DUSPs expressed in preadipocytes and show that dusp1, dusp8, and dusp16 are induced in response to TNFα. Of these, dusp8 and dusp16 are regulated by JNK in a negative feedback manner. Focus on kinase‐phosphatase relationships will increase our knowledge governing the inflammatory process that couples obesity to insulin resistance. Supported by NIH (5R21DK072067‐02).

Grant Funding Source

NIH 5R21DK072067‐02
Benzyl isothiocyanate (BITC) inhibits lipopolysaccharide (LPS)‐induced expression of iNOS and COX‐2 in murine macrophages
Authors: Lee yeo myeong , Seon Mi Ra , Cho Han Jin , Kim Jong‐Sang , Park Jung Han Yoon
Published: June 2021

BITC is found in high amounts in garden cress, papaya, and common Brassica vegetables, and some studies have shown that BITC may have potential as a chemopreventive agent in humans. The present study determined whether BITC inhibits lipopolysaccharide (LPS)‐induced inflammatory responses in Raw 264.7 macrophages. The treatment of cells with various concentrations of BITC led to a dose‐dependent reduction in NO and PGE2 production. Consistent with these findings, BITC inhibited LPS‐induced expressions of inducible nitric oxide synthase (iNOS) and cyclooxygase (COX)‐2 proteins and their corresponding mRNAs. BITC inhibited LPS‐induced degradation of inhibitory factor‐κB and translocation of p65 into the nucleus. Moreover, the LPS‐stimulated phosphorylation of extracellular signal regulated kinase (ERK)1/2 and was concentration‐dependently suppressed by BITC. Taken together, these results suggest that the anti‐inflammatory properties of BITC may result from inhibition of iNOS and COX‐2 protein expression through the downregulation of NF‐κB signaling.
Arginine supplementation improves insulin resistance in obese adolescents
Authors: Verbruggen Sascha , Coss‐Bu Jorge , Hsu Jean W‐Ch , Joosten Konrad , van Goudoever Johanness , Castillo Leticia
Published: June 2021

Arginine through a NO mediated mechanism improves insulin resistance in type 2 diabetes. To asses the effect of a short‐term (1 week) dietary arginine supplementation on insulin resistance in glucose intolerant obese adolescents, we conducted a randomized, cross‐over study in 12 subjects (16±1 yr.;BMI 36±5 kg/m2) receiving a one week diet period of a weight maintaining protein and energy intake, or an isocaloric arginine supplemented (21 gram/day) isonitrogenous diet. At the end of each diet period they received a primed, constant, 7 hr. tracer infusion of [1‐13C]Glucose, L‐[2H3]Leucine and [2H5]Glycerol. During the last 3 hours a hyperinsulinemic euglycemic clamp (HEC; 40 mU·m−2·min−1 insulin) was conducted. Arginine supplementation improved (p<0.05) insulin sensitivity from 0.3±0.2 to. 0.4±0.2 μmol ·kg−1 ·min−1/μU/mL. There was a trend towards improvement of non‐oxidative glucose disposal (3.5±3 vs. 4.5±2 mg.kg−1.min−1; p=0.06). However, there was no effect on endogenous glucose production (‐1.0±1 vs. ‐1.0±2 mg.kg−1.min−1; p=0.99). Arginine had no effect on protein breakdown or lipolysis. A one week oral arginine supplementation (21 gr/day) in obese adolescents improves insulin resistance.

NIH DK‐62363, T32 HD‐007445, Ajinomoto 3‐ARP; USDA/ARS 25337387;

Sophia Foundation for Scientific Research (SSWO).
Rat faecal microbiota composition associated with diet and phenotype
Authors: Jain Shalini , Levin Barry E , Pan Yuan‐Xiang , Miller Michael J
Published: June 2021

Recently, it has been proposed that the development of obesity is influenced by the relative proportion of Bacteroidetes and Firmicutes, the two major phyla of bacteria found in the gut. We determined the fecal microbiota composition in diet induced obesity (DIO) and diet resistant (DR) selectively bred Sprague Dawley (SD) rats on different diets. When fed a 31% fat, high energy (HE) diet, DIO rats become obese whereas DR rats remain lean. However, when fed standard chow, both DIO and DR remain lean. Fecal samples were analyzed from four different groups of rats; 1) DIO rats fed HE diet (DIO‐HE; n=5); 2) DIO rats fed standard chow (DIO‐Chow; n=5); 3) Diet resistant rats fed HE diet (DR‐HE; n=5); and 4) Diet resistant rats fed standard chow (DR‐Chow; n=5). The composition of gut microbiota was determined using 16s rDNA sequencing. There was no significant difference in the Bacteroidetes (p = 0.06) and Firmicutes (p = 0.07) levels between the DIO (obese) and DR (lean) phenotype when fed the HE diet. However, there was a significant difference in the Bacteroidetes and Firmicutes levels between the DIO and DR phenotype when on the standard chow diet (p=0.05) despite both groups having similar body weights. These changes indicate that diet and phenotype affects the gut microbiota composition and suggests that intentional manipulation of community structure may be useful for regulating obese phenotype. (Hatch project # ILLU‐698‐339)
Development, implementation and evaluation of a methodology to identify populations at risk for nutritional deficiency
Authors: Pachón Helena , Vesga Varela Andrea Liliana , Hyman Glen , Monserrate Rojas Fredy Alexander , Zapata Caldas Emmanuel
Published: June 2021

Local‐level data on the population's nutritional status are rarely available. For targeting, methods are needed to identify localities with populations at risk for nutritional deficiency. A method was developed that used biochemical (BIO), anthropometric (ANTHRO), intake, or socioeconomic (SES) data (with internationally recommended indicators & cutoffs) to identify localities at risk for iron, zinc, vitamin A or protein deficiencies in 11 Latin American countries. This method had several flaws: contradictory results when 2 ANTHRO indicators were used concurrently as recommended; difficulty in harmonizing BIO, ANTHRO & intake data (mostly at departmental level) with SES data (always at local level); & lack of data on recommended SES indicators. An improved method was developed: only BIO data are used to identify departments at risk for a specific nutrient deficiency; lacking BIO data, height‐for‐age Z score <‐2 is used to identify departments where populations are at risk for a nutritional deficiency (not nutrient‐specific); & SES data (poverty‐intensity index or proportion living in extreme poverty) are used in tandem with BIO or ANTHRO data to identify at‐risk localities within at‐risk departments. This method coupled with mapping software facilitates the identification of localities with nutritionally at‐risk populations. Funding: AgroSalud Project (CIDA), CIAT.
Modeling and pathway analysis of [U‐13C]starch conversion by a colonic microbiota
Authors: de Graaf Albert Arjaan , Venema Koen , Maathuis Annet , de Waard Pieter , Deutz Nicolaas
Published: June 2021

We study colonic microbial diversity, metabolic function of colonic microbes, the way diet affects metabolic fluxes, and how produced metabolites affect human health. Short chain fatty acids (SCFA) and especially butyrate, produced by the colonic microbiota, are important mediators of health and disease. Linking the biosynthesis of SCFA to specific gut microbial pathways holds promise for the development of prebiotics that specifically modulate these pathways in vivo. Therefore, 13C stable isotope labeling, NMR, and mathematical modeling of isotopomer distributions were employed to investigate colonic microbial metabolic pathway activities during [U‐13C]starch conversion in an in vitro model of the colon inoculated with a standardised fresh human fecal microbiota. GC‐MS, enzymatic methods and NMR at 500 MHz were used to identify microbial metabolites and to analyze their 13C contents.

The SCFA acetate, propionate and butyrate were determined as the principal starch degradation products. Quantitative insight in bacterial metabolic routes was obtained from analysis of 13C‐13C coupling patterns in 2D HSQC spectra that resulted from incorporation of intact 13C glucose backbone fragments in SCFA. These indicated that e.g. propionate was almost exclusively formed via the succinate pathway.

In conclusion, this stable isotope study underlines the importance of colonic bacterial SCFA metabolism.
Iron and Zinc Status are Not Affected by Soy Food Consumption in Women of Childbearing Age
Authors: Zhou Ying , Alekel D. Lee , Genschel Ulrike , Reddy Manju B
Published: June 2021

Many health benefits are attributed to soy food intake, but research on mineral balance is limited. Our hypothesis was that consumption of soy foods for 10‐wk would not affect iron or zinc status in women of childbearing age. Women (18‐28 y) non‐smokers without chronic disease, anemia, pregnancy, and/or irregular menstrual cycles were randomly assigned to either a soy foods (n=31) or control (n=32) group. Blood and 3‐day dietary records were collected prior to and after intervention. At baseline, there were no significant differences between the soy food and animal product groups in dietary intake or iron and zinc status. No significant changes were observed in serum ferritin, transferrin receptor, and hemoglobin concentrations between groups. Although serum iron (‐4.8 µg/dl) and body iron (‐0.53 mg/kg) declined in the soy group after intervention, the change was not significant either between the two groups or from baseline. Intervention also resulted in no change in zinc status as reflected by serum (7 µg/dL decrease in both groups) and urinary zinc in the soy compared with control group (17 vs ‐9 µg/g creatinine). Our results showed that incorporating 2 servings/day of soy foods containing about 20 g protein for 10‐wk had no detrimental effect on iron status or zinc status in women at‐risk for these mineral deficiencies.
Quercetin as an Inhibitor of Protein Glycation
Authors: Doran Emily , Hubbard Emily , Cheuk Anne , Vong Jennifer , Kruse Chase , Wing Barbara
Published: June 2021

The current study examines the ability of the compound quetrctin to inhibit the formation of gylcated proteins. This process of non‐enzymatic addition of sugars to proteins has been implicated as one of the contributing factors to the development of the complications of diabetes. In the model used in our laboratory the glucose was incubated with the protein lysozyme as a positive control for glycation. Quercetin was added as a possible inhibitor of glycation. Solutions were tested by spectophotometry for the presence of glycoprotiens. Initial results indicate that100mM and200mM concentrations of quercetin do inhibit the interaction between glucose and lysozyme. Thus querrcetin requires more detailed studies as a possible inhibitor of the long term complications of diabetes. This Research is supported by a grant from the Smith‐Glynn‐Callaway Medical Foundation.
Thromboxane Synthase Inhibition Blunts the Development of Pulmonary Hypertension and Vascular Remodeling in Hypoxic Neonatal Piglets
Authors: Hirenallur‐S. Dinesh K. , Haworth Steven T. , Gordon John B. , Rusch Nancy J.
Published: June 2021

The development of chronic hypoxia (CH)‐induced neonatal pulmonary hypertension (PH) is associated with increased production of thromboxane (TxA2) and an upregulation of L‐type calcium (CaL) channels. Thus, we hypothesized that TxA2 synthase inhibition may blunt the development of PH by mitigating the upregulation of CaL channels. Newborn piglets were exposed to 21 days of normoxia (N), CH or CH plus the TxA2 synthase inhibitor, furegrelate (oral 3 mg/kg, 3x daily). In vivo pulmonary vascular resistance index (PVRI) was 3.15 ‐fold higher in CH (104±1 WU) compared to N (33±1 WU) piglets. Furegrelate partially blunted the elevated PVRI in CH piglets (64±0.5 WU). Furegrelate also reversed the elevated transpulmonary pressure in isolated lungs of CH piglets by 66%, and blunted the overexpression of CaL channels and anomalous Ca2+‐dependent tone. Pulmonary arterial distensibility was decreased in lungs of CH compared to N piglets, indicative of vascular remodeling. Furegrelate partially restored distensibility to normal levels. Our findings suggest that pharmacological inhibition of TxA2 synthase blunts the development of hypoxia‐induced neonatal PH by preserving vascular function and structural integrity. Funded by NIH R01 HL83013
PIN1 regulates calpain‐mediated degradation of cyclooxygenase‐2 in endothelial cells
Authors: LIU TONGZHENG , Schneider Ryan A. , Shah Vaibhav , Likhotvorik Rostislav I. , Keshvara Lakhu , Hoyt Dale G.
Published: June 2021

Although the peptidyl‐proline isomerase, PIN1, regulates pro‐inflammatory factors, its role in cyclooxygenase‐2 (COX‐2) expression is unknown. Here, PIN1 was depleted in murine aortic endothelial cells (MAEC) by stable expression short hairpin RNA targeting PIN1 or a control construct. E. coli endotoxin (LPS) plus interferon‐gamma (IFN) induced COX‐2 4.7 times more in PIN1 knockdown (KD) than in control MAEC. KD did not affect induction of COX‐2 mRNA. Instead, KD increased stability of COX‐2 in cycloheximide‐treated cells. Previously, it was found that PIN1 KD reduced calpain activity and increased expression of the endogenous inhibitor, calpastatin. Here, induction COX‐2 was enhanced by the calpain inhibitor, MDL 28170, and loss of COX‐2 after cycloheximide was similar in KD cells and in controls treated with MDL 28170. COX‐2 in extracts of control MAEC was more sensitive to calpain‐1 digestion in vitro than COX‐2 from knockdown cells. Immunodepletion of PIN1 from control cell extracts did not affect calpain sensitivity, and no direct interaction between COX‐2 and PIN1 was seen. These results suggest that PIN1 regulates COX‐2 degradation mainly by its indirect effect on calpain via calpastatin. Thus, PIN1 may normally restrain expression of COX‐2 and other calpain substrates in endothelial cells. Modulation of PIN1 may significantly affect calpain‐dependent vascular activity and inflammatory diseases.
Purmorphamine stimulates osteoblastic differentiation of mesenchymal stem cells
Authors: Rosa Adalberto Luiz , Oliveira Fabiola Singaretti , Bellesini Larissa Serzut , Crippa Grasiele Edilaine , de Oliveira Paulo Tambasco , Beloti Marcio Mateus
Published: June 2021

This study aimed at evaluating whether purmorphamine stimulates osteoblastic differentiation of mesenchymal stem cells (MSCs) derived from human bone marrow. Primary MSCs were cultured in non‐osteogenic medium and characterized by fluorescence‐activated cell sorting (FACS) of a panel of surface markers. First passage MSCs were cultured for 7 days either in non‐osteogenic medium in presence of 2 µM purmorphamine and evaluated by FACS or in osteogenic medium in presence of 2 µM purmorphamine and gene expression of the osteoblastic markers Runx2, ALP, OC, OPN, and Msx2 evaluated by real‐time PCR. FACS of primary cultures revealed two distinct cell populations, the predominant MSCs (CD90+, CD73+, CD105+, CD166+, CD29+, CD13+, HLA class I+, CD146+, CD44+, CD14−, CD106−, CD31−, CD45−) and the fewer hematopoietic cells (CD34+). After 7 days, FACS demonstrated that purmorphamine treatment reduced surface markers expression while untreated cultures kept the primary MSCs immunophenotype profile. MSCs cultured in osteogenic medium supplemented with purmorphamine displayed upregulation of Runx2, ALP, OC, and Msx2 while OPN expression was not affected. FACS analysis suggests that purmorphamine treatment reduces mesenchymal stem cell population probably by stimulating osteoblastic differentiation as revealed by increased gene expression of osteoblastic markers. Grants from FAPESP and CNPq, Brazil.
In‐Vivo Functional Properties of the Fully Edited 5‐HT2C Receptor
Authors: Olaghere da Silva Uade Brimmy , Sanders‐Bush Elaine
Published: June 2021

In‐Vitro studies of RNA editing of the serotonin type 2C receptor (5‐HT2CR) have shown a pronounced loss of function in the fully edited isoform (VGV). Our goal is to evaluate functional coupling of the edited 5‐HT2CR in‐vivo using transgenic mice that solely express the VGV isoform. In order to accomplish this goal, we established selective activation of the receptor both in‐vitro and in‐vivo. Since pharmacological studies are not available for mice, we compared the relative affinity of antagonists for 5‐HT2A versus 5‐HT2C receptors, confirming that M100907 and SB206553 were relatively selective. Nearly a decade ago, our laboratory showed that RNA editing of the 5‐HT2CR to generate the VGV isoform silences constitutive activity of the receptor, eliminates high affinity agonist binding, and decreases agonist potency in cell culture. We now demonstrate, in‐vivo, a decrease in GTP‐sensitive high affinity binding from 5% in wild‐type mice to less than 0.1% in transgenic mice solely expressing the VGV isoform. We have also observed a marked 20‐fold increase in5‐HT2CR receptor density, as well as, an increased agonist and antagonist sensitivity in locomotor activity. (Supported by NIH grants T32MH065782 and R01MH34007)
Purification and partial characterization of a novel, non‐AT1, non‐AT2 binding site for angiotensins in the rat brain
Authors: Karamyan Vardan T. , Arsenault Jason , Klarskov Klaus , Shariat‐Madar Zia , Escher Emanuel , Speth Robert C.
Published: June 2021

The structure of the novel, non‐AT1, non‐AT2 binding site for angiotensins (Ang) (Brain Res. 2007, 1143: 83) is still unknown. We used Sar1‐p‐benzoyl‐L‐Phe8‐Ang II to specifically photolabel this protein in rat brain membranes. Photolabeled membrane pellet extracts were resolved by 7.5% SDS‐PAGE. The radioactive peak in the "Total" binding samples was 75‐80 kDa with negligible amounts of radioactivity at this point in corresponding "Non‐specific" samples. Gel cuts from the radioactive band from the "Total" samples were combined and radioactivity was extracted. The molecular weight of the semipurified binding site was not altered by glycopeptidase‐F indicating that the protein is not N‐glycosylated. Neither GTPgammaS nor GppNHP (at 50 µM) alter Ang II binding affinity for the binding site suggesting that it is not a G protein‐coupled receptor. The semipurified, radio‐photolabeled binding site was applied for 2D gel electrophoresis. The isoelectric point of the binding site was ~ 7.0. Cyanogen bromide hydrolysis of the protein gave 2 bands: 25 & 12.5 kDa, suggesting that there are 3 or more Met residues in this protein. Mass‐spectroscopic analysis of these digests should provide a partial amino acid sequence, revealing whether this is a known protein with a novel functionality, or if it is a novel brain protein. Supported by the Peptide Radioiodination Service Center of the University of Mississippi.
[beta]‐arrestin 1 mediates angiotensin II induced ubiquitination and down‐regulation of TRPV4
Authors: shukla arun kumar , kim jihee , ahn seungkirl , xiao kunhong , Liedtke wolfgang , lefkowitz robert j
Published: June 2021

β‐arrestins were originally discovered to desensitize activated G protein‐coupled receptors (GPCRs), also known as seven transmembrane receptor (7TMRs). It was subsequently found that β‐arrestins also mediate 7TMR internalization by bringing the activated receptors to clathrin‐coated pits. More recently, it has been discovered that β‐arrestins can also interact with and regulate ion channels and transporters. Using a proteomic approach, we identified TRPV4 as an interacting partner of β‐arrestin 1 after stimulation of cells with angiotensin II. Here we demonstrate that β‐arrestin 1 is recruited to TRPV4 upon angiotensin II stimulation in HEK‐293 cells and vascular smooth muscle cells. Surprisingly, angiotensin II stimulation leads to ubiquitination of TRPV4 which is abolished by β‐arrestin 1 siRNA. β‐arrestin 1 shows constitutive interaction with AIP4, a known ubiquitin E3 ligase for TRPV4, which is further increased upon angiotensin stimulation. Moreover, the interaction of TRPV4 and AIP4 is also significantly reduced by β‐arrestin 1 siRNA. These findings suggest that β‐arrestin 1 serves as an adaptor for AIP4 and thereby facilitates TRPV4 ubiquitination and down‐regulation. Taken together with the previous reports of analogous function of β‐arrestins for 7TMRs, this study suggests a novel role of β‐arrestins as a general adaptor for ubiquitin E3 ligases.
Alpha‐1A adrenergic receptor regulation of learning and memory in mice
Authors: Goldenstein Brianna , Boese Sarah , Schlosser Danielle , Knudson Chris , Carr Patrick , Haselton James , Perez Dianne , Doze Van
Published: June 2021

The hippocampus has been implicated in learning and memory and is targeted by the adrenergic system. Recently, it was shown that the alpha‐1A adrenergic receptor (α1A‐ARs) may regulate neuronal differentiation and neurogenesis. The function of α1A‐ARs is not well understood and in order to compensate for the lack of antibodies and specific ligands for this receptor, transgenic mice which over‐expressed the α1A‐AR with enhanced green fluorescent protein (EGFP) or constitutively active mutant (CAM) α1A‐AR were generated. Knockout (KO) α1A‐AR mice were also made. An immunohistochemical analysis of slices from each animal showed that CAM α1A‐AR mice had increased BrdU incorporation compared to normal and KO α1A‐AR mice. Increased hippocampal neuron counts were also found in CAM α1A‐AR mice compared to normal mice. This increase in neurons may affect learning and memory. Normal, CAM α1A‐AR, and KO α1A‐AR mice were tested using a multi‐component T‐maze, the Morris water maze and the Barnes maze. Overall, KO α1A‐AR mice displayed lower and CAM α1A‐AR mice higher cognitive ability in each model. α1A‐AR stimulation could offer a new strategy for treating the cognitive decline and neurodegeneration associated with aging and other age‐related neurological disorders. This study was supported by ND EPSCoR EPS‐0447679, NSF CAREER 0347259, NSF REU Site 0639227, NSF MRI 0619688, NIH P20 RR016741 and NIH RO1HL61438.
Evaluation of propofol effect in altering pentylenetetrazol induced seizure threshold in TNBS colitis models in rats
Authors: MEDHI BIKASH , Prakash Ajay , Prasad Byrav D S
Published: June 2021

To evaluate the role of propofol in altering pentylenetetrazol induced seizure threshold in colitis models in rats and its antagonist effect by thalidomide. Total 42 Wister rats of either sex were included in the study. Animals were divided into seven groups (n=6) control group (PTZ, 60mg/kg of b wt ), PTZ low dose group (40 mg/kg of b wt), propofol (5 mg/kg of body weight), propofol (5 mg/kg) + PTZ (40 mg/kg). TNBS induced colitis group (PTZ 40mg/kg b wt), TNBS colitis + propofol (5mg/kg) +PTZ (40mg/kg) and TNBS colitis + propofol (5mg/kg) +PTZ (40mg/kg) and thalidomide 150mg/kg orally for 14 days. The rats were given propofol (5 mg/kg), after 24 hour of I/P injection of propofol rats were subjected to PTZ (40 mg/kg) seizure susceptibility. Following parameters were assessed seizure assesment, morphological score, histological score and biochemical parameters (MPO, MDA, TNF‐α) The results showed onset of seizure was reduced in the propofol groups and TNBS colitis model which was significant different compared to control group (p<0.001). In severity score and duration of seizure was significant elevated in the propofol treated group and TNBS colitis model (p< 0.001). Thalidomide restored lipid peroxide malondialdehyde as well as reduction of myeloperoxidase and TNF‐α towards normal levels. Morphological and histological score were significantly reduced in thalidomide treated groups (p <0.001). Present study showed that propofol reduce the seizure threshold and increase duration of seizure in TNBS induce colitis model. (Supported by PGIMER, Chandigarh, India.)
Fast calcium responses along endothelium of arteriolar networks during blood flow
Authors: Bagher Pooneh , Davis Michael J. , Segal Steven S.
Published: June 2021

To study endothelium‐specific calcium signaling in vivo we utilized the superfused (pH 7.4; 34 °C) cremaster muscle preparation in anesthetized male transgenic mice expressing the calcium sensor GCaMP2 under endogenous transcriptional regulation of connexin40 (Cx40). Endothelial calcium waves have been characterized as a transient fluorescence increase initiated by ACh that travels bidirectionally along arterioles at 116 µm/s and decays within 1 mm. Here we report a novel "fast calcium response (FCR)" that travels at mm/s along arteriolar networks in the direction of convection during blood flow. To determine whether FCR was blood flow‐dependent, an arteriole (diameter, 30‐50 µm) was occluded upstream or downstream from the site of abluminal ACh microiontophoresis (1 µm pipette tip; 500 ms, 500 nA). The FCR was eliminated when blood flow ceased (n=4), whereas removing the occluder restored the FCR along with blood flow. Redirecting flow into a side branch also redirected the FCR. These findings demonstrate that focal release of a vasoactive substance adjacent to an arteriole can activate entire networks of arteriolar endothelial cells through gaining access to the flow stream. Support: NIH HL041026, AR048523.
Age‐related effects of varying ammonia concentrations on hematophysiological variables in broiler chickens,
Authors: OLANREWAJU HAMMED AKANDE , PURSWELL JOSEPH L. , COLLIER STEPHANIE D.
Published: June 2021

This study examined the response of different aged birds of the same genetic strain exposed to ammonia at set concentrations on blood physiological variables under environmentally controlled conditions. The experiment consisted of 4 levels (0, 25, 50 and 75 ppm) of ammonia concentrations and 4 different ages (1‐day, 7‐days, 14‐days and 21‐days) arranged in a complete randomized factorial design. Venous blood samples were collected at the end of each 7 days of ammonia exposure. Partial pressure of CO2 (pCO2), pH, hematocrit (Hct), and hemoglobin (Hb) increased significantly (P = 0.05), whereas partial pressure of O2 (pO2), bicarbonate (HCO3−), and K+ decreased with increasing ammonia concentration compared with 0 ppm. In addition, pO2, pCO2, HCO3−, Hct, Hb, Na+, and anion gap (Angap) increased significantly (P = 0.05), while pH, glucose, and corticosterone decreased as bird's age increased. Ammonia × age interactions were observed for pH, anion gap, and HCO3−. There was significant main effect of age but not that of ammonia on plasma corticosterone concentrations. The main effect of age was more pronounced than that of ammonia on examined variables. These conditions improved as the age of birds increased from 1‐d to 21‐d old. It was concluded that different aged birds of the same genetic strain respond differently to set ammonia concentrations and younger birds have a more intense reaction to the ammonia than older birds.
Mathematical modeling of metabolism‐perfusion matching in a microvascular network
Authors: Roy Tuhin K , Joyner Michael J , Secomb Timothy W
Published: June 2021

Microvascular blood flow regulation has important implications for optimization of metabolism‐perfusion matching in physiological and pathophysiological conditions such as exercise and sepsis. The purpose of this theoretical study is to assess the impact of various regulatory factors on metabolism‐perfusion matching. A microvascular network model including myogenic, shear, and metabolic flow regulation was developed and utilized to obtain estimates of oxygen extraction under control conditions as compared to a 20% global increase in blood flow accompanied by a 50% increase in oxygen consumption. The exercise state was simulated with preserved flow regulation and sepsis was simulated with impaired metabolic flow regulation. For the values of parameters used, overall oxygen extraction was found to be 28% higher in exercise compared to control values as opposed to only 23% higher in sepsis. This study demonstrates the impact of impaired flow regulation on oxygen delivery and highlights microvascular perfusion abnormalities as a putative mechanism for organ failure in critically ill patients.
THE C‐TERMINAL GLOBULAR DOMAIN OF FIBRINOGEN γ CHAIN INDUCES ENDOTHELIAL BARRIER DYSFUNCTION AND MICROVASCULAR LEAKAGE VIA A RHOA‐DEPENDENT MECHANISM
Authors: Guo Mingzhang , Daines Dayle , Tang Jing , Perrin Rachel M. , Shen Qiang , Takada Yoshikazu , Wu Mack H. , Yuan Sarah Y.
Published: June 2021

Fibrinogen degradation products have been associated with endothelial dysfunction in inflammation. There is evidence that the C‐terminal globular domain of fibrinogen γ chain (γC) binds to endothelial cells, however, its functional impact on vascular barrier properties remains poorly understood. We examined the effects of recombinant γC on microvascular endothelial permeability using in vivo and in vitro models. The results showed that γC increased hydraulic conductivity and albumin permeability in rat mesenteric microvessels. In addition, γC induced endothelial monolayer barrier dysfunction in a dose‐dependent manner, concomitantly with endothelial Rho GTPase activation, myosin light chain phosphorylation, and stress fiber formation. Blockage of RhoA signaling with Y27632 inhibited the γC‐induced endothelial barrier dysfunction and microvascular leakage. Likewise, depletion of RhoA with siRNA prevented the hyperpermeability response. In contrast, the full‐length fibrinogen protein and the C‐terminal globular domains of fibrinogen αE and β chains did not cause significant changes in endothelial barrier function or microvascular permeability. Taken together, the results suggest that γC exerts a direct effect on the microvascular endothelium causing hyperpermeability via a RhoA‐dependent signaling pathway. Supported by NIH RO1 HL‐073324, HL‐061507 and HL‐070752.
Stimulation of Epac/Rap1 pathway deactivates hyperpermeability after its onset in PAF‐induced acute inflammation in vivo
Authors: Iwahashi Toru , Kim David D , Lal Brajesh K , Durán Walter N
Published: June 2021

Epac (exchange protein activated by cAMP) regulates endothelial cell‐cell adhesion. We tested the hypothesis that stimulation of Epac can deactivate the inflammatory hyperpermeability caused by PAF (platelet‐activating factor). We used integrated optical intensity (IOI) as the index of microvascular permeability and examined tissues by intravital microscopy and image analysis. Two minutes after finishing the administration of 10−7M PAF, we applied the Epac‐specific cAMP analogue, 8‐(4‐chloro‐phenylthio)‐2′‐O‐Methyladenosine ‐3',5′‐cyclic monophosphate (8‐CPT) at 10−5M to stimulate Epac. PAF increased net IOI from baseline to a mean ± SEM value of 142.1 ± 5.3 in the mouse cremaster and from baseline to 70.7 ± 8.9 in hamster cheek pouch. 8‐CPT reduced these values to 68.5 ± 8.2 and 34.2 ± 9.7, respectively (P <0.05). Administration of 10−5M L‐NMMA or 10−6M ABT‐491 (PAF receptor blocker) after application of PAF did not inhibit PAF‐induced hyperpermeability. We conclude that stimulation of Epac deactivates PAF‐induced pro‐inflammatory hyperpermeability. (Supported by 5R01 HL070634 and 1RO1 HL088479).
Enzymatic endothelial glycocalyx degradation increases arterial‐venous oxygen exchange in the heart
Authors: Brands Judith , Post Mark , Vink Hans , Teeffelen Jurgen Van
Published: June 2021

A relatively thick endothelial glycocalyx forms the true interface between blood and the vascular wall, and thereby limits microvascular volume available for perfusion and exchange. In previous studies we have shown that hyaluronidase treatment increases coronary microvascular blood volume. We now hypothesize that acute functional or enzymatic reduction of glycocalyx volume increases microvascular exchange capacity. In the current study the effect of enzymatic glycocalyx degradation on myocardial oxygen exchange was assessed in goats (n=5). Blood samples were collected simultaneously from great cardiac vein and aorta, and blood flow was measured in the LAD. The measurement was done in control and after hyaluronidase treatment. Hyaluronidase treatment increased arterial‐venous oxygen difference significantly from 7.0 ± 0.2 to 8.3 ± 0.3 ml/100ml, which was associated with a reduced coronary flow from 66.5 ± 9.1 to 46.4 ± 2.9 ml/min/100gr tissue (P<0.05). Rate‐pressure product and oxygen consumption remained unaltered. In conclusion, acute enzymatic degradation of glycocalyx volume increases arterial‐venous oxygen exchange, consistent with our previous finding on the effect of hyaluronidase treatment on microvascular blood volume. Supported by NHF grants 2005T037 and 2003B181.
“In vivo” mechanical compliance of the initial lymphatic vessels in rat diaphragm
Authors: Negrini Daniela , Moriondo Andrea , Boschetti Federica
Published: June 2021

The lymphatic system originates from the interstitial tissue with initial lymphatic capillaries, whose wall is devoid of smooth muscle cells. In these vessels, lymph formation and progression is sustained by cyclic tissue stresses transmitted to the vessel lumen according to the mechanical properties of the vessel walls, which are at present still unknown. Therefore, we aimed at measuring in vivo the mechanical properties of initial lymphatics belonging to the diaphragmatic network in anesthetized rats. The diaphragm was exposed and a lymphatic vessel was localized via stereomicroscopy. Vessel segments were occluded at both ends and injections of 4.6 nL saline were performed, while measuring lymphatic pressure (Pl) by micropuncture. At each injection, Pl step increase was followed by stress relaxation (τ= 2.9 ± 0.2 (SE) sec). For incremental volume steps < 35 % of initial segment volume (Vo), two vessel populations were identified with mechanical compliance (C) of 40.8 ± 4 nl/cmH2O and 2.8 ± 0.4 nl/cmH2O. For volume steps from 35 to 100 % and greater than 100% of Vo, C was equal in both populations and averaged 4.5 ± 0.6 nl/cmH2O and 62.4 ± 7.4 nl/cmH2O, respectively. In conclusion, the existence of two populations of initial lymphatic vessels with distinct mechanical behaviour suggests a differential recruitment of lymphatic vessels to cope with the varying draining requirements of the tissue