bluuebunny/crossref_metadata_2025 huggingface dataset:
G Protein‐Coupled Receptor (GPCR) Signaling Pathways in Beta‐Cells
Authors: Wess Jürgen , Ruiz de Azua Inigo , Gautam Dinesh , Scarselli Marco , Rosemond Erica , Cui Yinghong , Guettier Jean‐Marc
Published: June 2021

G protein‐coupled receptors regulate many aspects of beta‐cell function. To assess the physiological relevance of the different beta‐cell G protein signaling pathways in vivo, we generated a series of mutant M3 muscarinic acetylcholine (ACh) receptors (M3 mAChRs) which are unable to bind the endogenous ligand, ACh, but can activate different classes of G proteins when treated with the pharmacologically inert compound, clozapine‐N‐oxide. Studies with transgenic mice selectively expressing these modified M3 mAChRs in pancreatic beta‐cells have shed new light on the in vivo roles of distinct G protein signaling pathways in beta‐cell function.

The M3 mAChR is a prototypic Gq‐coupled receptor. Recent data suggest that strategies aimed at enhancing signaling through beta‐cell M3 mAChRs may be beneficial for the treatment of type 2 diabetes by promoting glucose‐stimulated insulin secretion. To identify regulatory proteins that reduce M3 mAChR signaling in pancreatic beta‐cells, we used the MIN6 murine insulinoma cell line as an in vitro model system. By using siRNA technology, we identified several proteins, including different receptor kinases and RGS proteins, that acted as negative regulators of M3 mAChR function in MIN6 cells. Given the key role of M3 mAChR signaling in beta‐cell function, it is likely that this approach will lead to promising new targets for the treatment of type 2 diabetes.
n‐3 Fatty Acids Are Neuroprotective After Cerebral Hypoxia‐Ischemia In Rodent Models
Authors: WILLIAMS JILL J , Bazan Nicolas , Ten Vadim S , Vannucci Susan J , Mastropietro Christopher , Deckelbaum Richard J
Published: June 2021

We questioned if acute administration of n‐3 FA decreases brain injury after hypoxia‐ischemia (H/I), and if so, what mechanisms are involved. Juvenile Wistar rats and adult and neonatal C57/BL6 mice underwent right carotid artery ligation followed by exposure to 8% O2, and received n‐3‐rich FA emulsion or saline (controls). 72h (rats) and up to 48h (mice) post H/I, animals were euthanized. Brain sections were stained with TTC, H&E, and Fluoro Jade to analyze injury. FA content was determined by GLC. Mouse brains were analyzed by GC mass spectrophotometry for neuroprotectin D1 (NPD1). Cerebral blood flow (CBF) was measured using Laser‐Doppler flowmetry.

Injury scores were 0.42±0.62 (n‐3) vs. 2.00±0.83 (control rats) (p < 0.005) with similar results in adult and neonatal mice. In controls, but not in n‐3, there was a ~25% decrease in each FA/g rat brain in right hemispheres, indicating brain edema. Injured areas display cell loss, pyknotic nuclei and degenerating neurons. In whole mouse brain, NPD1 increased at 8h post H/I in n‐3 compared to controls (2.0±0.4 vs. 0.9±0.4 pmole/mg protein, p<0.05). There were no n‐3 related differences in CBF. Thus, acute administration of n‐3 FA is protective against H/I induced brain infarction in 3 rodent models. Neuroprotection relates to an early response to cerebral H/I by NPD1 and to prevention of brain edema.

Grant Funding Source

NIH‐NHLBI
Camelina sativa oil and lipid changes in postmenopausal women
Authors: Campbell Christina Gayer , Syndergaard Sarah , Filipowicz Rebecca , Thorland William G.
Published: June 2021

Camelina sativa is a novel plant source of alpha‐linolenic acid (ALA) with a similar fatty acid profile to flaxseed. A double‐blind parallel‐arm pilot study investigated the lipid lowering effects of daily consumption of camelina sativa oil for 6 weeks. Twenty overweight women (mean ± SD; 28.0 ± 2.6 kg/m2) were randomized to one of two groups: 2T/d camelina oil (CO)(8g ALA/d) (n=10) or 2T/d sunflower oil (SO)(0g ALA/d)(n=10). Subjects consumed an ad libitum diet; test foods provided as baked goods. Weight significantly increased over time for both groups (CO: 70.8 ± 4.9 kg vs. 75.3 ± 7.8 kg; SO: 77.2 ± 10.2 kg vs. 79.4 ± 10.7 kg)(P<0.05). A significant decrease for total cholesterol (TC) (CO: 4.5%; SO: 7.8%) and high‐density lipoprotein (HDL) (CO: 22%; SO: 18%) was observed for both groups (P<0.05). A decrease (SO: 3.3%; P>0.05) in plasma low‐density lipoprotein (LDL) was found whereas an increase of 7% (p>0.05) occurred for the CO group. Results suggest that changes in the lipid profile are attainable with a plant‐based polyunsaturated fatty acid despite significant weight gain. The safflower oil was more effective at producing positive lipid changes than the camelina sativa oil. Future research is warranted with more controlled feeding interventions to minimize weight gain. Funding was provided by Biobased Institute of Montana (USDA‐CREES 2005‐34510‐15726).
Choline intake, plasma riboflavin and the PEMT G5465A genotype predict plasma homocysteine in folate deficient Mexican American men with the MTHFR 677TT genotype
Authors: Caudill Marie A , Dellschaft Neele , Hinkis Sabrina , Ivanov Alexandre A , Nash‐Barboza Susan , Randall Katharine E , Jackson Brandi , Solomita Gina N , Vermeylen Francoise
Published: June 2021

We previously showed that provision of the folate RDA and either 300, 550, 1100, or 2200mg choline/day resulted in diminished folate status and a tripling of plasma total homocysteine (tHcy) in men with the MTHFR 677TT genotype However, the substantial variation in tHcy within the 677TT genotype implied that several factors were interacting with this genotype to affect homocysteine. This study sought to identify the main predictors of wk‐12 plasma tHcy, alone and together with the MTHFR C677T genotype (29 TT, 31 CC), using linear regression analysis. A basic model explaining 82.5% of the variation (i.e., adjusted R2 =0.825) was constructed. Within the 677TT genotype, serum folate (P=0.005) and plasma riboflavin (P=0.002) were strong negative predictors (inversely related) explaining 12% and 15% of the variation in tHcy; whereas choline intake (P=0.003) and serum creatinine (P<0.001) were strong positive predictors explaining 19% and 25% of the variation. None of these variables, except creatinine (P=0.021), correlated with tHcy within the 677CC genotype. When creatinine was not in the model, the PEMT 5465G>A variant predicted lower tHcy (P<0.001); an effect confined to the MTHFR 677TT genotype. Thus, in folate deplete men, several factors with roles in one‐carbon metabolism interact with the MTHFR C677T genotype to affect plasma tHcy.
Associations of food insecurity, socioeconomic status, and type 2 diabetes among Hispanic and non‐Hispanic adults in the NHANES 1999‐2004
Authors: Fitzgerald Nurgul , Jang Seo Yeon
Published: June 2021

Hispanics and people with low socioeconomic status (SES) have higher rates of type 2 diabetes (T2D). We examined whether food insecurity and low SES are independently related to T2D among Mexican Americans and non‐Hispanic whites (NHW). About 2955 Mexican Americans and 6363 NHWs, 20‐84yr, from the nationally representative NHANES 1999‐2004 were included in the analyses. Compared to highly food secure people, those with marginal or very low food security were more likely to have T2D after adjusting for education, employment, poverty, gender, age, acculturation, and ethnicity (p<0.05). Following further adjustment for other risk factors (obesity, physical activity, dietary and alcohol intakes, smoking, family history diabetes, and co‐morbidities), only those with very low food security remained to be more likely to have T2D (OR 1.84, CI 1.02‐3.31). When the two ethnic groups examined separately, very low food security became a stronger correlate of T2D among NHWs (OR 3.53, CI 1.58‐7.87); this association was attenuated among Mexican Americans. Low education and unemployment were related to higher likelihood of having T2D among Mexican Americans (p=0.050) but not among NHWs. These results suggest that associations of food insecurity and SES with T2D vary between Mexican Americans and NHWs, and this may require different prevention efforts tailored to the needs of each racial/ethnic group.
Potato consumption reduced oxidative stress and inflammatory damage in humans
Authors: Kaspar Kerrie L , Park Jean Soon , Mathison Bridget , Brown Charles R , Navarre Roy , Chew Boon P
Published: June 2021

Potatoes contain phenolic acids, anthocyanins and carotenoids; however, their potential benefits in immune modulation and against oxidative stress and inflammation are unknown. Free‐living healthy male participants (18‐40 yr; n=12/group) ate 150 g of cooked White (WP), Yellow (YP) or Purple (PP) flesh potatoes once a day for 6 wk in a single‐blinded study. A 3‐day dietary recall was obtained. Blood was collected at baseline and wk 6 to analyze total antioxidant capacity, 8‐hydroxydeoxyguanosine (8‐OHdG), protein oxidation, lipid peroxidation, C‐reactive protein, IL‐6, TNF‐α, IFN‐γ, lymphoproliferation, NK cytotoxicity and phenotypes. Compared to participants fed WP, those fed PP (P<0.1) or YP (P<0.05) had lower plasma 8‐OHdG while subjects fed PP had lower (P<0.1) C‐reactive protein. Plasma IL‐6 also was lower (P<0.07) in PP. The population of total Tc cells was lower (P<0.05) in PP, while subjects fed YP tended to have lower (P>0.1) B and NK cell subpopulations and higher total T cell populations than those fed WP. Similarly compared, subjects fed PP tended (P>0.1) to have higher B and NK cell, and lower total T cell populations. Therefore, consumption of pigmented potatoes reduced inflammation and DNA damage, and modulated immune cell phenotype in male subjects. Support: Washington State Potato Comm.

Grant Funding Source

Washington State Potato Commission
The Euplotes crassus selenoproteome
Authors: Lobanov Alexey V , Turanov Anton A , Klobutcher Lawrence A , Hatfield Dolph L , Gladyshev Vadim N
Published: June 2021

The hypotrichous ciliate Euplotes crassus genome has been analyzed for the presence of selenocysteine‐containing genes. Metabolic labeling of E. crassus revealed selenoprotein pattern that was similar to that observed in human HEK 293 and algal Ostreococcus tauri cells. Several selenoproteins were identified, including a new selenoprotein belonging to the Pfam09409 family (designated ep22 for Euplotes 22 kDa protein) with homologs in other ciliates, Tetrahymena and Paramecium, and in a primitive animal, Nematostella. Sec was encoded by UGA codon in all selenoprotein genes, which also had eukaryotic SECIS elements in their 3′‐UTRs. Four selenoprotein genes had UGA codons encoding both Sec and Cys, whereas in other selenoprotein genes, UGA only encoded Sec. Consistent with the use of UGA codon for both Sec and Cys insertion, we identified the corresponding Sec and Cys tRNAs containing UCA anticodon. This finding shows that the previous prediction of a single Cys tRNAGCA serving UGA, UGC and UGU codons in Euplotes is incorrect. Analysis of the Euplotes selenoproteome expands our understanding of selenoprotein evolution and genetic code.
One codon – two amino acids.
Authors: Turanov Anton A , Lobanov Alexey V , Fomenko Dmitri E , Morrison Hilary G , Sogin Mitchell L , Klobutcher Lawrence A , Hatfield Dolph L , Gladyshev Vadim N
Published: June 2021

In ciliated protozoa Euplotes crassus, cysteine (Cys) is encoded by three codons, UGA, UGU and UGC. UGA is a stop signal in the universal genetic code, and this codon can also code for the 21st amino acid, selenocysteine (Sec), in all three domains of life. Metabolic labeling of E. crassus with 75Se revealed specific incorporation of radioactive Se into several proteins. We found that in this organism the UGA codon specifies insertion of both Sec and Cys, and that the dual use of this codon can occur within a single gene. We studied a Euplotes protein, thioredoxin reductase 1 (eTR1), which has seven in‐frame UGA codons. Our experiments demonstrated that Sec was only inserted into the classical Sec site in eTR1, whereas other UGA positions were not served by SECIS for Sec insertion and instead supported termination of translation in mammalian cells. LC‐MS/MS sequencing revealed eTR1 peptides containing Cys in positions 63, 68, 208, and 270, which are encoded by UGA codons, whereas peptides containing Sec at these positions were not detected. Thus, our data show that E. crassus utilizes UGA for insertion of both Cys and Sec, establishing it as the first known organism that utilizes one codon to code unambiguously for two different amino acids.
Cooking of spinach influences the isomeric profile of beta‐carotene but does not significantly affect its micellarization
Authors: O'Sullivan Laurie , Galvin Karen , Jiwan Marvin , Aherne‐Bruce Aisling , O'Brien Nora
Published: June 2021

The influence of processing on the micellarization and potential bioavailability of carotenoids from spinach has been previously investigated; however, few studies have looked at the effects of processing on β‐carotene (BC) isomer micellarization. Spinach is often cooked by boiling prior to consumption. The aims of the present study were, first, to investigate the effects of domestic cooking (by boiling) on the isomeric profile of BC in spinach and, second, to assess the effects of cooking on the micellarization of BC isomers. Raw and cooked spinach were homogenized and subjected to an in vitro digestion procedure. The micelle fraction was isolated by ultracentrifugation. Carotenoids were extracted, saponified and analyzed by HPLC. All‐trans BC was the predominant BC isomer in raw and cooked spinach. Following cooking, the percentage contribution of 9‐cis to total BC was significantly (P<0.001) increased and the % contribution of all‐trans BC to total BC was significantly reduced (P<0.001). Micellarization of BC isomers ranged from ca. 20.3 to 60.9 %. Although cis BC isomers in both raw and cooked spinach were more efficiently micellarized when compared with the all‐trans isomer, this trend was not significant. Cooking increased BC isomer micellarization, however this increase was not significant. Our findings indicate that cooking spinach alters the isomeric profile of BC, however, efficiency of micellarization is not significantly affected. This work is funded by Science Foundation Ireland.
Over‐expression of Peroxisome‐Proliferator‐Activated Receptor‐Gamma Co‐Activator 1 Beta decreases cellular triglyceride content and secretion in HepG2 Cells
Authors: Johnson Diana Roshek , Brinker Anita M , Dixon Joseph L
Published: June 2021

The Peroxisome‐Proliferator‐Activated Receptor‐Gamma Co‐Activator 1 Beta (PGC1β) is a co‐activator of several transcription factors involved in lipid metabolism and very low density lipoprotein (VLDL) secretion. Therefore, PGC1β may be a master regulator of lipid metabolism in liver. To study this hypothesis, human liver HepG2 cells were infected with adenoviruses containing PGC1β or β‐galactosidase (control), and cells and medium were extracted to determine changes in cellular lipids. A wide array of lipids was measured using liquid chromatography/mass spectrometry (LC/MS/MS) using an Applied Biosystems 4000Q Trap. PGC1β over‐expression caused significant decreases in liver cellular neutral lipids. There was a 61.5±3.9% decrease in triacylglycerides (TAG), 40.9±6.7% decrease in diacylglycerides (DAG), and 52.9±3.8% decrease in monoacylglycerides (MAG). During this period, over‐expression of cellular PGC1β decreased the secretion of neutral lipids: 39.7±4.9% decrease in TAG, 60.2±4.1% decrease in DAG; MAG were not detectable in HepG2 medium. There were no significant changes in any of the cellular phospholipid concentrations or species between control cells and cells treated with PGC1β. Our studies show that over‐expression of PGC1β greatly influences neutral lipid metabolism in human liver HepG2 cells.

Grant Funding Source

NIH, Rutgers University
Hepatic gene profiling prior to puberty reveals greater female predominant expression of lipogenic genes
Authors: Shankar Kartik , Ronis Martin J , Badger Thomas M
Published: June 2021

We evaluated changes in transcriptomic profiles in livers of pre‐peubertal rats using Affymetrix microarrays. Female Sprague‐Dawley rats were fed a phytoestrogen free, AIN‐93G‐based diet containing casein as the sole protein source from gestation day 4 and offspring were weaned onto the same diet. Male and female offspring were sacrificed on PND25 and hepatic gene expression assessed using the Rat 230 2.0 GeneChip microarrays. Data were analyzed using GeneSpring Gx7.3 and Ingenuity Pathway Analyses and transcripts that changed ±1.8‐fold (p <0.05) were enriched. Expression of 60 transcripts was significantly altered between genders. Among these, genes involved in lipid biosynthesis and elongation, adiponutrin (11‐fold), fatty acid synthase (4‐fold), ATP citrate lyase (2.9‐fold), ELOVL6 (5‐fold), acetyl‐coA carboxylase (2‐fold) and thyroid hormone responsive Spot 14 (2‐fold) were increased in females compared to males. Importantly, mRNA expression of the master lipogenic factor SREBP‐1 was increased 2.2‐fold in female liver. In addition, genes regulating fatty acid oxidation, Cpt1a, CYP4A14 and Slc22a5 and cholesterol transporters Abcg5, Abcg8 were lower in female livers (p<0.05). These results indicate significant gender differences in SREBP‐1 regulated lipogenic gene expression prior to divergence in major sex hormones at puberty. Support ARS‐CRIS 6251‐51000‐005‐00D.
Docosahexaenoic acid supplementation from mid‐pregnancy through parturition influenced breast milk fatty acid composition at 1 month post‐partum in a double‐blind randomized controlled trial in Mexico
Authors: Imhoff‐Kunsch Beth C , Stein Aryeh D , Villalpando Salvador , Martorell Reynaldo , Ramakrishnan Usha
Published: June 2021

Omega‐3 polyunsaturated fatty acids (PUFAs), including docosahexaenoic acid (DHA), are essential for neural development and accumulate extensively in the fetal and infant brain. Studies have shown that DHA concentration in human milk (HM) increases with maternal supplementation with fish oil during pregnancy and/or lactation, but the benefits of prenatal supplementation with DHA alone are unknown. We investigated the effect of prenatal DHA supplementation on PUFA concentrations in HM at 1 mo post‐partum (PP). In a double‐blind RCT in Mexico, 1040 pregnant women received 400 mg DHA or placebo daily from 18‐22 weeks' gestation through parturition. Fatty acid (FA) concentrations (% of total FA) were determined using gas liquid chromatography in HM obtained from 174 women at 1 mo PP. Women receiving DHA had higher mean HM DHA and alpha‐linolenic acid (ALA) concentrations, expressed as % of total FAs, compared to placebo at 1 month PP. Mean HM DHA concentrations in DHA and placebo groups were 0.20 (SD=0.06) and 0.17 (SD=0.07), respectively (Δ=0.02, 95% CI=‐0.04, ‐0.004), and mean ALA concentrations were 1.38 (SD=0.47) and 1.24 (SD=0.46), respectively. Concentrations of other FAs such as eicosapentaenoic and arachidonic acid did not differ between groups (P>0.05). Prenatal DHA supplementation increased concentrations of DHA and ALA in HM 1 month PP, providing a mechanism through which breastfed infants might benefit.

Grant Funding Source

NIH (HD043099)
Risk factors for delayed onset of lactogenesis among northern California primiparous women
Authors: Nommsen‐Rivers Laurie Ann , Cohen Roberta J , Chantry Caroline J , Peerson Janet M , Dewey Kathryn G
Published: June 2021

Our objective was to determine factors associated with delayed onset of lactogenesis (delayed OL)‐defined as onset of stage II lactogenesis after 72 h postpartum (pp)‐among a multi‐ethnic sample of primiparae who initiated breastfeeding (N=415). Prevalence of delayed OL was 42%. Variables associated with delayed OL (P < .10, unadjusted) were greater maternal age, education level, and body mass index (BMI), Cesarean delivery, birth weight > 3600 g, lower Apgar scores, edema pp, formula intake > 60 ml over first 48 h, and lack of nipple pain at 24 or 72 h pp. A hierarchical series of multiple variable logistic regression models wasere used to examine independent effects of the above variables on the risk of delayed OL. The final model included the following significant (P < .05, adjusted) variables: maternal age, BMI, birth weight and formula use. There was a strong association between higher BMI and increased risk of edema (P < .0001); edema was significant in an alternate final model excluding BMI. DFurthermore, delayed OL increased the risk of excess neonatal weight loss (loss > 10% of birth weight): prevalence was 34.2% versus 7.8% among infants of mothers with vs.ersus without delayed OL (P< .0001). We conclude that delayed OL is common among this sample of California primiparae and is associated with both biological and behavioral factors. Supported by a grant from the Department of Health and Human Services, Maternal Child Health Bureau.
L‐Cysteine supplementation lowers blood glucose, CRP, MCP‐1 and inhibits NFkB and Akt activation in liver of Zucker diabetic rats
Authors: Jain Sushil K. , Velusamy Thirunavukkarasu , Rains Justin , Croad J L , Bull R
Published: June 2021

Many proteins, including insulin contain free sulfhyryl groups that are considered to be pivotal for their biological functions. L‐Cysteine is a semi‐essential amino acid with a free sulfhyryl group. This study examined the hypothesis that L‐cysteine supplementation can lower insulin resistance, glycemia and markers of vascular inflammation in type 2 diabetes using Zucker diabetic fatty (ZDF) rats.

Starting at the age of 6 wks, ZDF rats were supplemented orally (daily gavages for 8 more wks) with saline‐placebo (D) or L‐cysteine (LC, 1mg/KgBW), and fed with a high calorie diet. ZDF rats of 6 wks age without any supplementations were considered as baseline (BL). Blood was collected by heart puncture using anesthesia from BL, D and LC supplemented rats.

D rats showed elevated levels of fasting blood glucose, HbA1, CRP, MCP‐1, when compared to BL rats where there is no onset of diabetes. LC supplementation significantly lowered blood glucose (18%, p=0.05), HbA1 (8%, p=0.02), CRP (23%, p=0.02), MCP‐1 (32%, p=0.01) and HOMA index of insulin resistance (25%) when compared to D. There were no changes in blood GSH and lipid peroxidation levels between D and LC. While LC did not change blood hematocrit or levels of transaminases, however it did lower alkaline phosphatase (29%, p=0.01) levels in comparison to D. Western blotting analyses of liver showed increased activation of NFkB (50% pNFkB) and Akt (20%, pAkt) in D when compared with BL. LC supplementation reversed these effects (17% pAKT, 18% pNFkB). Thus, L‐cysteine supplementation can lower glycemia and markers of vascular inflammation, apparently by preventing NFkB activation, in ZDF rats.
Composite Phenotypes of Interrelated Lipoprotein Subclasses Independently Predict Diet‐Induced Changes in Plasma Lipids and Lipoproteins
Authors: Chiu Sally , Krauss Ronald M
Published: June 2021

There is considerable interindividual variation in lipid and lipoprotein responses to diets aimed at reducing risk for cardiovascular disease. We used unsupervised principal component (PC) analysis to test whether composite measures of plasma levels of individual VLDL, LDL, and HDL subfraction mass as measured by analytical ultracentrifugation identified phenotypes that predicted changes in plasma lipids and lipoproteins in response to low‐fat, high‐carbohydrate diets in 548 healthy non‐obese men. Three PCs accounted for 72% of the overall variance in lipoprotein fractions at baseline (high fat). The largest (PC1, 42%), represented a combination of increased small LDL and reduced large LDL. PC1 was significantly (p<0.0001) associated with % reductions in triglyceride, LDL cholesterol (C), and apoB, and increases in HDLC. PC2 (19%), which represented increased mass of small HDL, IDL, and medium and large LDL, was associated (p<0.001) with % reductions in both LDLC and HDLC. PC3 (11%) consisted of increased small and large HDL, together with reduced medium LDL, and was associated (p<0.0001) with % HDLC reduction. The lipoprotein subfraction phenotypes identified by PC analysis may represent the effects of distinct metabolic pathways that contribute independently to variation in response to dietary interventions for improving plasma lipid profiles. (support: National Dairy Council).
Stress‐suppressing model explains mediation of food insecurity in the effect of poverty reduction program on wellbeing and distress
Authors: Jalal Chowdhury Sufi Bhakta , Frongillo Edward A
Published: June 2021

Poverty alleviation programs for the extreme poor improve participants' economic status and may also affect other important outcomes that are seldom evaluated. A program targeted to the extreme poor by BRAC, a development organization in Bangladesh, has been successful in significantly alleviating extreme poverty and improving wellbeing, and reducing distress and food insecurity. We hypothesized that reduced food insecurity played a major role in mediating the effect of the program on wellbeing and distress. A non‐equivalent pre‐ and post‐test quasi‐experimental design was used to test the hypotheses. Linear mixed random‐intercept models and path analysis was used to test the hypothesis. Data were collected from a random sample of 209 women (110 program and 99 control) across 27 villages of 3 northern districts in 2002 and 2005‐2006. Reduced food insecurity was found to be a substantial mediator of the program's effect on other outcomes. Approximately 53% of the effect of the program on distress and 66% of the effect of the program on wellbeing was mediated through food insecurity. The results of this study are highly important as this is a large‐scale program already extended to half of the country. Findings will contribute in designing, judging the cost‐benefit and cost‐effectiveness, and in garnering support for the expansion of such programs.
Prospective analysis of national nutrition policy advancement in Viet Nam
Authors: Frongillo Edward A , Lapping Karin , Studdert Lisa J
Published: June 2021

Improved understanding of how to advance national nutrition policy is greatly needed. We studied prospectively why and how national nutrition policy advanced in Viet Nam between 2006 and 2008. Using a participant‐observer, change‐agent model, we examined strategies used, factors that shaped policy advancement, and how factors and strategies interacted to advance policy. Data were collected using questionnaires, informant interviews, program visits, document reviews, and documentation of key events. For analysis, we created a chronology of events, examined strategies and actions used and their results by event, coded interviews, and tabled factors applying Shiffman's 2007 framework. We used three analytical methods to discern why and how policy advanced. Developing a cohesive policy community with well‐defined internal and external frames that were articulated through a series of high‐profile policy windows was critical. Key strategies capitalized on cultural motivations and values. High‐profile events facilitated by the international community were important. Results show that nutrition policy advancement is possible if key factors are favorable, thoughtful strategies are used, and the international community supports country leaders. The participant‐observer, change‐agent model was successful in both contributing to policy advancement and documenting it. Funded by the World Bank.
Development and evaluation of an intervention to promote in‐home ready‐to‐use food (RUF) consumption in a supplemental feeding program for undernourished and stunted children in western Uganda
Authors: Ickes Scott Bradley , Chamiec‐Case Linda , Jilcott Stephanie B , Ammerman Alice S
Published: June 2021

Overview

Supplemental feeding programs (SFP) have reported low frequency and overall intake of ready‐to‐use food (RUF) rations, which may affect growth outcomes. Efforts are needed to improve feeding frequency and intended RUF dosage.

Methods

Semi‐structured interviews were conducted among caregivers (n=32), health center staff (n=8), and RUF producers (n=8) to identify the barriers to in‐home RUF consumption. Following analysis of the interviews, feeding messages to promote RUF consumption were developed and used in the 10 week program curricula. To evaluate the effect of these messages on RUF intake, dietary data were obtained using two 24‐hour recalls conducted within 10 days at caregiver homes or at health clinics. Group means (SE) of RUF feeds per day and frequency of RUF inclusion in total meals were compared between Pre‐Intervention (n=18) and Feeding Education (n=18) cohorts.

Results

The Feeding Education group reported 1.9 (0.2) RUF feeds per day, compared with 1.3(0.2) in the control group (t=‐2.3, p<0.05). The frequency of RUF inclusion in meals was 0.57 (0.06) in the EC group and 0.39 (0.05) in the control (t=‐2.1, p<0.05).

Conclusion

Messages to promote RUF consumption can have positive effects on RUF dosage and feeding frequency, which may contribute to improved energy intake and growth outcomes among malnourished children in SFPs.

Grant Funding Source

GlaxoSmithKline UNC‐Duke Student Research Grant; Carolina Center for Public Service Entrepreneurship in Public Service Fellowship
Mitochondria Dynamics Abnormalities in Alzheimer Disease
Authors: Wang Xinglong , Su Bo , Perry George , Smith Mark A , Zhu Xiongwei
Published: June 2021

Mitochondrial dysfunction has long been implicated in the pathogenesis of Alzheimer disease (AD). Mitochondrial function is highly dependent on mitochondria morphology dynamics, which are regulated by mitochondria fission/fusion proteins, i.e., DLP1/Fis1 for fission, and Mfn1/Mfn2/OPA1 for fusion. In this study, we found that levels of DLP1, OPA1, Mfn1 and Mfn2 were reduced while the level of Fis1 was increased in AD hippocampus. More importantly, all these proteins accumulated in the soma and depleted in neuronal processes in AD neurons. To explore the consequence of changes in these mitochondrial fission/fusion proteins, we manipulated the expression of them in M17 cells and primary hippocampal neurons in a way that mimicked their expression changes in AD and found that despite their differential effects on mitochondrial morphology, they all caused reduced mitochondrial density in cell periphery or neuronal processes. In an attempt to address the potential cause of abnormal mitochondrial dynamics, we found that ADDLs treatment not only led to mitochondrial fragmentation but also led to mitochondrial depletion from neuronal processes. Interestingly, the overexpression of DLP1 rescued ADDLs‐induced abnormal mitochondrial distribution while overexpression of OPA1 restored ADDLs‐induced mitochondrial fragmentation. Most importantly, we found ADDLs‐induced mitochondrial depletion correlated with reduced spine density and synaptophysin density and puncta number in neuronal processes, which could be effectively reversed by overexpression of DLP1 through repopulation of neuronal processes with mitochondria. Taken together, we suggest an impaired regulation of mitochondrial dynamics caused mitochondrial dysfunction and neuronal dysfunction in AD neurons.
Expression of Aquaporin 4 mRNA during Acute Stage of Traumatic Brain Injury in Rats
Authors: Hao chunyan , Duan hubin , Li shuzhen
Published: June 2021

Backgroud and purpose

The aquaporin 4 (AQP4) water channel is abundant in rats brain and this protein plays a major role in the control of water flow within the central nervous system. This study is to investigate the expression of AQP4 mRNA and its correlation with brain edema during the acute stage of traumatic brain injury (TBI) in rats.

Methods

56 adult male Wistar rats were randomized into the TBI group and control group.TBI group were again stratified into 7 sub‐groups according to the time after injury (0.5,2,6,12,24,48 and 72 h).Severe injury models (2.5‐3.0atm ) was produced with the fluid percussion brain injury (FPI) technique in anesthetized adult rats equipped with a closed cranial window.At each time spots, the water contents in the brain tissues were measured by wet‐to‐dry weight ratio and the expression of AQP4 mRNA was assessed by semi‐quantitative RT‐PCR.

Results

The expression of AQP4 mRNA was up‐regulated 0.5 hours after injury,progressively enhanced from 2h to12 h, peaked at 24 h(P<0.05) and remained a high level at 72 h.A similar change was noted for water contents in brain tissues. The expression of AQP4 mRNA was positively correlated with brain water contents(r=0.095,P<0.05).

Conclusion

The changes in AQP4 mRNA expression may be closely related to formation and progression of brain edema during the acute stage of severe traumatic brain injury.AQP4 may be playing an important role in brain edema after severe TBI.
Protein kinase CK1αLS mediates hydrogen peroxide stimulated vascular cell proliferation and intimal hyperplasia
Authors: Stone James R. , Panchenko Mikhail P. , Silva Nilsa , Dombkowski David M. , Preffer Frederic I. , MacGillivray Thomas E.
Published: June 2021

Protein kinase CK1αLS and its substrate hnRNP‐C constitute a novel nuclear signaling pathway in vascular cells that is activated by mitogenic levels of hydrogen peroxide and is upregulated in intimal hyperplasia and atherosclerosis. The role of CK1αLS in vascular cell proliferation was assessed using retrovirally‐delivered shRNA to lower the expression of the kinase. Knock down of CK1αLS inhibited proliferation of human umbilical vein endothelial cells, human coronary artery endothelial cells, and human coronary artery smooth muscle cells (CASMC). Knock down of CK1αLS also abolished hydrogen peroxide stimulated proliferation of CASMC with accumulation of these cells in cell cycle phase G0/G1 by flow cytometry. Human internal thoracic arteries in culture developed smooth muscle cell proliferation, increased proteoglycan deposition, and intimal hyperplasia. All of these pro‐atherosclerotic changes were inhibited by knocking down CK1αLS. These findings indicate that protein kinase CK1αLS positively regulates vascular cell proliferation and intimal hyperplasia and specifically mediates vascular smooth muscle cell proliferation in response to low physiologic levels of hydrogen peroxide.
VE‐cadherin‐mediated signaling regulates microtubule dynamics
Authors: Komarova Yulia , Garcia Alexander N , Malik Asrar B.
Published: June 2021

Disruption of adherens junctions (AJs) in endothelial cells by pro‐inflammatory stimuli such as thrombin induces microtubule (MT) destabilization (Gorovoy et al., J Biol Chem. 2005 280: 26533‐42). Here we determined the signaling mechanism regulating MT dynamics by VE‐cadherin homophilic adhesion. We assessed MT dynamics in human microvascular endothelial monolayers by expressing YFP‐tagged End Binding (EB)‐1 protein, which binds preferentially to the growing ends of MTs. Formation of AJs inhibited MT growth by increasing MT catastrophe frequency whereas disruption of AJs by calcium switch or using VE‐cadherin blocking antibody induced MT persistent growth. We used calcium switch model to address the signaling mechanism downstream of VE‐cadherin adhesion in regulating MT dynamics. We observed that disruption of AJs induced the release of calcium from endoplasmic reticulum and activation of protein phosphatase 2B (PP2B). Using PP2B autoinhibitory peptide we demonstrated that PP2B induced de‐phosphorylation of EB3 (but not EB1 or EB2), the accumulation of EB3 at the MT growing ends, and MT growth. We conclude that signaling activated by VE‐cadherin adhesion regulates MT dynamics through the PP2B‐EB3 axis, and may thereby regulate endothelial cells shape and motility.
Low Serum Insulin‐Like Growth Factor 1 Potentiates Atherosclerotic Plaque Development in APOE‐/‐ Mice: Potential Mechanism of Accelerated Atherosclerosis in Aging
Authors: Shai Shaw‐Yung , Kelly James , Higashi Yusuke , Sukhanov Sergiy , Delafontaine Patrice
Published: June 2021

Aging is associated with reduced serum insulin‐like growth factor‐1 (IGF‐1) and an increased risk of atherosclerosis, but whether lower IGF‐1 plays a causative role is unknown. To determine the relation between serum IGF‐1 levels and atherosclerotic burden independent of age, a congenic mouse strain with 20% reduction in IGF‐1 (C3H.B6‐6T [6T]) was bred into the ApoE ‐/‐ background to obtain C3H.6T +/+/ ApoE ‐/‐ (6T/E) mice to mimic the clinical state of low IGF‐1. Serum IGF‐1 levels in 6T/E mice were similar to those of 6T (6T/E: 208.81 + 58.33 ng/ml vs 6T: 221.41 + 58.33 ng/ml, p=0.6); but lower than those of ApoE ‐/‐ (354.49 + 76.33 ng/ml, p<0.01). 8‐wk old 6T/E, 6T and ApoE ‐/‐ mice were fed normal chow (NC) or western diet (WD) for 12 wks and aortic plaque burden quantified. Plaque area was higher in 6T/E mice than ApoE ‐/‐ whether on NC or WD (% total plaque/aorta, 6T/E, NC: 7.54 + 2.21, ApoE ‐/‐, NC: 4.91 + 3.18, p<0.018; 6T/E, WD: 14.87 + 8.81, ApoE ‐/‐, WD: 8.71 + 3.13, p=0.0005). Body weight gain and blood pressure in 6T/E and ApoE ‐/‐ mice were the same. Thus, in this novel mouse model that mimics age‐related low IGF‐1, a decrease in serum IGF‐1 is accompanied by a diet‐independent increase in aortic atherosclerosis. These results are consistent with epidemiological data indicating that lower IGF‐1 levels are associated with an increased risk for ischemic heart disease, and strongly suggest causality.

Supported by NIH HL 70241
Fibroblast Growth Factor Receptor 3 (FGFR3) modulates Paneth cell lineage allocation through a β‐catenin/Tcf‐4 dependent signaling mechanism
Authors: Brodrick Brooks Barrett , Vidrich Alda , Buzan Jenny , Bradley Leigh , Cohn Steven M.
Published: June 2021

Fibroblast growth factor receptors are receptor tyrosine kinases that mediate cellular growth and differentiation during normal gut development and injury‐repair. Robust FGFR3 expression is restricted to the lower portion of the crypt where stem cells reside during normal intestinal development. Activation of the β‐catenin/Tcf‐4 complex is crucial for maintaining the intestinal stem cell compartment and for Paneth cell lineage allocation/differentiation. Previously we showed that FGFR3‐/‐ mice have fewer intestinal crypts, have a reduction in the total number of clonogenic stem cells, and decreased β‐catenin/Tcf‐4 activation. FGFR3‐/‐ mice at post‐natal days 14 and 21 display both a significant reduction in the number of intestinal Paneth cells and a reduction in mRNA levels of Paneth cell products. Since Paneth cell lineage allocation/differentiation is dependent upon β‐catenin/Tcf‐4 activation, we examined whether FGFR3 signaling can directly modulate Paneth cell specification. Caco2 cells treated with FGF9, a FGFR3 ligand, or co‐transfected with a constituitively active mutant (K650E) of FGFR3 had sustained β‐catenin/Tcf‐4 transcriptional activity. FGF9 increased mRNA levels of two Paneth cell markers, lysozyme and human defensin 5. These findings suggest that FGFR3 signaling can directly modulate Paneth cell specification possibly through the β‐catenin/Tcf‐4 signaling complex.
Loss of beta‐catenin Accelerates N‐nitrosodiethylamine–Induced Hepatocarcinogenesis by Activating Phosphoinositide 3‐Kinase (PI3K)/Akt Signaling Pathway
Authors: Zhang Xufeng , Tan Xinping , Monga Satdarshan S
Published: June 2021

β‐catenin, the pivotal component of canonical Wnt pathway, plays an important role in liver regeneration, growth and cancers. To further address the role of β‐catenin in liver carcinogenesis, we used a hepatocyte specific β‐catenin knockout (Ko) mouse model. 5 μg/g body weight of N‐nitrosodiethylamine (DEN), a well‐known hepatocarcinogenic agent, was injected i.p., at the age of 14 days to induce tumors in both Ko and wild type (WT) mice. Intriguingly, rather than being protected from carcinogenesis, β‐catenin Ko mice were more susceptible to DEN. In the Ko, liver tumors appeared more rapidly and were also larger and more in numbers. The Ko livers after DEN also showed a considerable increase in the number of proliferating cells by PCNA staining. We identified upregulation of platelet‐derived growth factor receptor‐α (PDGFRα), glycogen synthase kinase 3β (GSK3β), GSK3β (Ser 9) and phospho‐Akt (Thr 308) in Ko livers. Also phospho‐Akt (Thr 308) was exclusively upregulated in tumors in Ko livers as seen by immunohistochemistry. In conclusion, β‐catenin Ko livers were more prone to carcinogenesis, secondary to PI3K/Akt activation, thus accounting for earlier onset of tumor and an overall robust disease.
miRNA Profiles of Canine Lymphoid Cell Lines
Authors: Uhl Elizabeth W. , Suter Steven , Antes Travis , Tompkins S. Mark
Published: June 2021

miRNAs are very highly conserved across species, however their role in the regulation of cell proliferation and differentiation in normal and neoplastic canine tissues is unknown. Lymphoma is one of the most common malignant tumors of the dog, accounting for about 25% of all cancers. The purpose of this study was to determine whether miRNA arrays developed for human samples can be used to characterize canine tumors. Human cancer microRNA qPCR arrays were used to establish the miRNA profiles of 4 canine lymphoid cell lines: 2 of B cell origin and 2 of T cell origin. Over ninety percent of the human miRNA probes detected canine miRNAs. miRNA analysis of canine lymphoid cell lines identified 12‐15 miRNAs that are either 10‐fold over or 10‐fold under expressed compared to normal canine peripheral blood mononuclear cells. In addition, the relative expression of six miRNAs known to be upregulated in human chronic lymphoid leukemia was determined in the four canine lymphoid cell lines, and expression of miR‐20a and miR‐92 were found to be markedly increased in the cell lines. The results of this study indicate that miRNAs are highly conserved between humans and dogs, and that miRNA profiles have potential predictive, diagnostic and prognostic potential in canine lymphoma.
Reproducibility of miRNA Expression by Proliferating C2C12 Cells Using RT‐PCR Array
Authors: Chen Yongxin , Gelfond Jonathan , McManus Linda M. , Shireman Paula K.
Published: June 2021

RT‐PCR array is a new technique to study microRNA (miRNA) expression; however, minimal data regarding intra‐ and inter‐assay reproducibility are available. This study examined the variability of the TaqMan MicroRNA Array (Applied Biosystems) in the evaluation of RNA obtained from proliferating murine myoblast C2C12 cells. The same RNA was used for two separate reverse transcription (RT) reactions and the RT products were used in duplicate arrays. The log2 expression levels of most genes (66%) in the four replicates had a standard deviation (SD) below 0.5 with median SD=0.2. When cycle threshold (CT) exceeded 32 there was a significant SD increase from 0.23 to 1.07, thus, genes with an average CT>32 (129/381) were excluded. With CT<32, 82% of miRNA exhibited a SD<0.2. The endogenous control RNA Mammu6 correlated well in the four replicates with a coefficient of variation of 0.5% (SD<0.1) and average CT=19.6. Thus, the variability due to different RT reactions on replicate samples was low and approximately equal. We estimate that the platform can detect a fold change of 2 with 99% power and a type I error rate 0.0005, assuming 4 biological replicates under 2 conditions given the gene has an SD of 0.1. In conclusion, this real‐time PCR based array method provided highly reproducibly estimates of miRNA expression by proliferating myoblasts in culture. Supported by the Veterans Administration and USPHS HL07446 and HL074236.
Association of Hexokinase and Voltage Dependent Anion Channel in Cardiac Hypertrophy
Authors: Saran Varun , Saeedi Ramesh , Wambolt Richard , Allard Micheal F.
Published: June 2021

Glycolysis is accelerated in pathologic cardiac hypertrophy but the cellular mechanism(s) responsible are not yet fully understood. Binding of hexokinase (HK), a key enzyme in control of glycolysis in heart and other tissues, to mitochondrial voltage dependent anion channels (VDAC) is viewed as a major factor responsible for insulin‐induced glucose uptake in skeletal muscle and acceleration of glycolysis in malignant neoplasms. We hypothesized that binding of HK to VDAC is increased in pathologically hypertrophied heart muscle cells and hearts. Binding of HK to VDAC was assessed by co‐immunoprecipitation and immunoblot analysis of lysates from cultured H9c2 cells hypertrophied by exposure to arginine vasopressin (AVP) and from hypertrophied hearts in mice with an abdominal aortic constriction (AAC). Cells unexposed to AVP and sham‐operated mice served as Controls. AVP‐treated H9c2 cells and hearts from mice with an AAC were hypertrophied 20 to 30% with rates of glycolysis 30 to 50% greater than Controls. Co‐immunprecipitation experiments indicated that association of HK with VDAC was significantly increased in hypertrophied H9c2 cells and in hypertrophied mouse heart tissue. This supports the concept that binding of HK to VDAC is increased in the setting of pathologic cardiac hypertrophy and may play a role in the acceleration of glycolysis observed.

Supported by the Canadian Institutes of Health Research
Phenotype‐transformed interstitial cells in canine and human myxomatous mitral valves express tryptophan hydroxylase 1
Authors: Orton Christopher , Disatian Sirilak , Lacerda Carla
Published: June 2021

Tryptophan hydroxylase 1 (TPH1) is the limiting enzyme for peripheral serotonin synthesis. Expression of TPH1 by valve interstitial cells (VIC) could implicate an autocrine serotonin signaling mechanism in the pathogenesis of myxomatous valve disease (MVD). The study objectives were to: 1) determine expression of TPH1 in canine and human myxomatous mitral valves, and 2) identify VIC phenotype expressing TPH1.

METHODS

Expression of TPH1 was determined by immunoblot (IB) and immunofluorescence microscopy (IFM). TPH1 expression was co‐localized with markers of VIC phenotype transformation; α‐smooth muscle actin (α‐SMA) and embryonic non‐muscle myosin (SMemb).

RESULTS

Canine and human myxomatous mitral valves strongly expressed TPH1 on IB and IFM analyses. α‐SMA and SMemb exhibited distinctly different expression patterns that were similar in canine and human myxomatous valves. TPH1 expression co‐localized with SMemb+ phenotype VIC, but not α‐SMA+ phenotype VIC.

CONCLUSIONS

TPH1 expression by canine and human myxomatous mitral valves implicates autocrine serotonin signaling in the pathogenesis of MVD. TPH1 expression is associated with VIC phenotype transformation. Autocrine serotonin signaling could form the basis of a unifying hypothesis for the pathogenesis of spontaneous MVD and serotonergic drug‐induced valvulopathy. Research supported by the Heart to Heart foundation.
Bone Marrow derived stem cell subpopulations that show specific homing abilities
Authors: Curcio Francesco , Moretti Massimo , Toller Matteo , Spelat Renza , Ferro Federico , Bottecchia Moira , Bergamin Natasha , Beltrami Antonio Paolo , Cesselli Daniela , Beltrami Carlo Alberto , Ambesi‐Impiombato Francesco Saverio
Published: June 2021

Bone marrow derived stem cells (BMSCs) are able to generate many different cell types. Understanding biological significance and molecular mechanisms underlying adult stem cell trafficking seems crucial. Currently though, the real potential of BMSCs is not yet fully accepted and data reported after injection in uninjured animals are contradictory. This may be due to the use of total BMSCs, which may contain at variable rate and percentage different subpopulations with specific homing properties.

We isolated from the BM of C57B1/6‐Tg(UBC‐GFP)30Scha/J adult mice, 4 different populations according to molecular markers or culture methods: CD105+, MSC, floaters (D. Trisler's method) and buffy coat. After extensive immunophenotyping and microarray analysis, they have been injected in the tail vein of C57B1/6 wildtype mice. Animals have been sacrificed 2 days later and different tissues harvested. In each tissue, the presence of GFP+ cells has been analyzed by PCR and immunohistochemistry. Here we present data indicating not only specific homing of the subpopulations in different tissues, but even preferential localization within each tissue.