bluuebunny/crossref_metadata_2025 huggingface dataset:
Relation of Copper Toxicosis in Dogs and Wilson Disease to the Appearance of a Small Copper Carrier (SCC) in Blood Plasma and Urine
Authors: Flynn Stephen , Dalphin Matthew , Tellez Miguel , Fieten Hille , Weldy Scott , Muchenditsi Abigael , Lutsenko Svetlana , Linder Maria
Published: June 2021

The main route for copper excretion in mammals is via the bile, little exiting through urine. In Wilson disease, biliary excretion is prevented by lack of an active form of ATP7B, which permits copper entry into bile. This leads to copper accumulation and severe liver toxicosis. Dogs tend to have the same problem, with much higher liver copper levels than other mammals. Many die of liver copper toxicosis. As in Wilson disease, Atp7b‐/‐ mice excrete high levels of copper in urine. Excretion is in the form of a 1.5 kDa SCC. We now have evidence that the same SCC exists in the blood plasma of humans, mice and dogs. Normally, it is bound to larger proteins, but large amounts of free SCC occur in the plasma of Atp7b‐/‐ mice, and in some dogs. SCC can be released from larger proteins by EDTA and accounts for ~10% of the total copper in human plasma. SCC is also released by EDTA in plasma from dogs and mice, including those with “free” SCC. FPLC‐SEC preliminary data suggest SCC is riding on proteins >200kDa. Dog ceruloplasmin is also unusual, eluting as a much larger protein, although of normal size in SDS‐PAGE. We conclude that SCC may offer an alternative route for copper excretion that can be mobilized in some mammals in response to liver copper accumulation, and that the study of dog plasma may reveal additional information about copper metabolism not detected as easily in other species.
The Zinc Transporting Network is a Central Component of Dysregulation in Breast Cancer
Authors: Chandler Paige , Kochupurakkal Bose , Lopez Veronica , Alam Samina , Soybel David , Kelleher Shannon
Published: June 2021

Breast cancer is classified into 3 major molecular subtypes (Luminal, HER2 and Basal), which differ in prognosis and therapeutic options. Numerous reports find that zinc (Zn) hyperaccumulates in breast tumors; however, the mechanisms responsible for Zn accumulation and the functional consequences of zinc accumulation are not understood. We tested the hypothesis that mechanisms responsible for Zn accumulation are subtype‐specific and that distinct patterns of sub‐cellular Zn distribution reflect the malignant phenotype of the cancer.

Microarray gene expression revealed a subtype‐dependent pattern of Zn transporter expression in human breast tumors and cancer cell lines. Breast cancer cells accumulated significantly more Zn (p<0.05) than nonmalignant cells. Protein profiling of all 24 Zn transporters revealed that T47D luminal cells (LC) had greater expression of vesicular Zn importers and more vesicularized Zn pools, while MDA‐MB‐231 basal cells (BC) overexpressed the cytoplasmic Zn binding protein metallothionein. A primary difference was that LCs overexpressed the key Zn transporter ZnT2 and hyperaccumulated 2.5‐fold more Zn than BCs which did not express ZnT2. ZnT2 overexpression in MDA‐MB‐231 cells accumulated Zn in vesicles, altered morphology, arrested cells in G1 of the cell cycle and reduced invasion by ~80%.

Zn accumulation in breast tumors is due to subtype‐specific alterations in the Zn transporting network. This suggests that these alterations may underlie differences in the malignant phenotype observed in certain breast cancer subtypes.
Cranberries Modulate Postprandial Metabolism in Obese Participants with Type 2 Diabetes
Authors: MORRIS STACY , BETTS NANCY , LYONS TIMOTHY , BASU Arpita
Published: June 2021

Background

Cranberries have been shown to lower risks of type 2 diabetes, and improve features of the metabolic syndrome, and lower postprandial glucose in a few clinical studies. Purpose: We investigated the postprandial effects of dried cranberries vs. control (bananas) in obese participants with type 2 diabetes (T2DM), in a randomized crossover trial. Methods: Blood draws and vascular measurements were conducted at fasting, 1, 2, 3 and 4 hours (h), following consumption of a fast‐food style high‐fat breakfast (50g fat, 766 kcal). Results: Analyses of our study data (n=25; BMI (kg/m2) (mean±SD) =39.5±6.5; Age (years) =56±6) revealed that postprandial changes in HDL‐cholesterol tended to be lower in the cranberry vs. control up to 4h (p<0.1), while postprandial changes in glucose were significantly lower in the cranberry vs. control at 1 & 2h (p<0.05). Among vascular parameters, no significant differences were noted in mean arterial blood pressure and systolic and diastolic blood pressure, as well as in small and large artery elasticity indices.

Conclusions

Overall, cranberries were shown to exert modest but notable effects in high‐fat breakfast induced postprandial glucose and lipid levels in participants with T2DM. These findings define the role of whole cranberries in postprandial blood glucose management and need further investigation.
Meta‐Regression Analysis of the Effect of Trans Fatty Acids (TFAs) on LDL‐Cholesterol
Authors: Vincent Melissa , Allen Bruce , Liska DeAnn , Dourson Michael , Haber Lynne
Published: June 2021

The Food and Drug Administration (FDA) published a Federal Register notice tentatively determining that partially hydrogenated oils (PHOs, the primary dietary source of industrial TFAs – iTFAs) are no longer generally recognized as safe (GRAS) when used in food. This determination was based on the conclusion that there is no threshold intake for iTFA that would not increase an individual's risk of coronary heart disease, based primarily on research showing iTFAs increase LDL‐cholesterol (LDL‐C) at high levels of intake. A meta‐regression analysis was conducted on the relationship between changes in TFA intake and changes in plasma LDL‐C, building on an evidence map designed to capture all relevant data. The purpose of this analysis was to describe the dose‐response in the range of the human data, particularly the low‐intake region. An advantage of the weighted regression (i.e., meta‐regression) is greater accuracy, since studies with smaller variance are given more weight. A variety of different mathematical expressions were fit to the data to determine which one best fit the data. Variability was very large, particularly at low TFA intake, indicating TFA intake is not the primary determinant of LDL‐C levels at low TFA intake. No significant differences were found in the mathematical fit of models across a wide range of shapes, from very flat initial slope to very steep initial slope. Thus, the mathematical modeling cannot inform our understanding of the shape of the dose‐response curve even in the range of human data, and determine whether a threshold exists. Instead, as is generally the case, one needs to rely on a mode of action evaluation.

This work was sponsored by the ILSI North America PHO Task Force.
Role of Heat Shock Proteins in Modulating ECM Deposition
Authors: Leavitt Jeffrey , Whalen Rachel , Veraldi Kristen , Binder Robert , Yates Cecelia
Published: June 2021

Objective

The hallmark feature of systemic sclerosis (SSc) is excessive accumulation of extracellular matrix (ECM) in skin and other affected organs that leads to fibrosis. The accumulation of ECM is caused by pathologically activated fibroblast that releases collagen, fibronectin, and other ECM components. Strong evidence suggests transforming growth factor beta (TGF‐β) plays a pivotal role in fibroblast activation and induces excessive ECM production. In addition, heat shock protein (HSPs) have been implicated in SSc such that pharmacological inhibition of hsp90 blocks the pro‐fibrotic effect of TGF‐β. In this study we evaluate the role of hsp90 and hsp70 on TGF‐β signaling and ECM production in human dermal fibroblasts.

Method

Expression of hsp90 and hsp70 were quantified by immunoblot and immunofluorescence microscopy following TGF‐β induction of dermal fibroblasts. The effect of hsp90 and hsp70 inhibition on TGF‐β‐induced ECM expression was analyzed in dermal fibroblasts.

Results

Expression of hsp90 and hsp70 increased in a TGF‐β‐ dependent manner 48 hours after treatments following a marginal, proteasomal‐dependent decrease at 24 hours. Inhibition of hsp90 by 17‐allylamino‐17‐demethoxy‐geldanamycin (17‐AAG) blocked the effects of TGF‐β on fibronectin and alpha‐smooth muscle actin and myofibroblast differentiation. In turn, there was an increase in antifibrotic matrix decorin.

Conclusion

Our data suggest a critical role for hsp90 in TGF‐β signaling. We propose the use of hsp90 inhibitors as potential antifibrotic treatments. Our data suggest a critical role for hsp90 and hsp70 in TGF‐β signaling. Blockade of TGF‐β‐mediated myofibroblast differentiation by 17‐AAG suggests that hsp90 inhibitors merit additional study as novel antifibrotic agents.
Interaction of the Protein Kinase C‐related Kinases with the TPα and TPβ isoforms of the T Prostanoid Receptor: Role in Thromboxane‐induced Neoplastic Responses
Authors: Mulvaney Eamon , O'Sullivan Aine , Hyland Paula , Kinsella B
Published: June 2021

In humans, thromboxane (TX) A2 signals through the TPα and TPβ isoforms of the T prostanoid receptor (TP). There is increasing evidence of a role for TXA2 and TPα/TPβ in neoplasms including in prostate, colon and bladder cancer. Mechanistically, we previously identified protein kinase C‐related kinase (PRK) 1 as an interactant of TPα/TPβ, a functional interaction essential for TPα/TPβ‐induced prostate cancer (PCa) cell migration. Moreover, agonist‐activation of TPα/TPβ induced phosphorylation of histone H3 at Thr11 (H3Thr11), a recognized epigenetic marker of androgen‐induced chromatin remodelling. As PRK1/PKN1 is a member of a wider subfamily including PRK2/PKNγ and PRK3/PKNβ which, like PRK1, are also widely implicated in cancer progression, herein the interaction of TPα/TPβ with PRK2 and PRK3 was investigated. Both PRK2 and PRK3 were confirmed to interact with TPα and TPβ, interactions dynamically regulated in response to TP‐agonist activation. Agonist‐activation of the TPs also led to activation of all three PRKs. Furthermore, PRK1 and PRK2, but not PRK3, were found to interact with TPα and TPβ in PC‐3 cells, and targeted disruption of PRK1 and PRK2, but not PRK3, impaired TP‐mediated H3Thr11 phosphorylation, cell proliferation, migration and invasion in the PCa cell line. Considering the critical roles of TPα/TPβ and of the PRKs in cancer progression, the discovery of these distinct, yet regulated, interactions is likely to be functionally significant, not least in cancer, but also in other pathophysiological settings where aberrant PRK and/or TP expression or function are implicated. Funded by the Irish Cancer Society, Health Research Board Ireland and the Movember Foundation (PCI12KIN, MRCG/2012/3).
Eicosapentaenoic Acid Prevents Interstitial Cardiac Fibrosis in a Mouse Model of Hypertensive Heart Disease
Authors: Eclov Julie , Shearer Gregory , O'Connell Timothy
Published: June 2021

Heart failure with preserved ejection fraction (HFpEF) is ~50% of all heart failure (HF) diagnoses, however, standard HF therapies show no efficacy in HFpEF. Omega‐3 polyunsaturated fatty acids, eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), improve outcomes in coronary heart disease and HF with reduced ejection fraction, but whether this benefit extends to HFpEF is unclear. Here, we examined the effect of EPA and DHA on remodeling following transverse aortic constriction (TAC) in mice. TAC induced concentric hypertrophy, interstitial fibrosis, and diastolic and systolic dysfunction similar, in several respects, to remodeling in HFpEF. Diastolic dysfunction correlated with the extent of interstitial fibrosis and hypertrophy after TAC (R2=0.39, P=0.02; R2 = 0.34, P=0.04, respectively), but systolic dysfunction did not (R2=0.19, P = 0.14; R2=0.07, P=0.40). Interestingly, blood levels of EPA (n=58, P=0.04), but not DHA (n=58, P=0.63), inversely correlated with interstitial fibrosis after TAC. However, EPA, unlike DHA, was not incorporated into cardiac fibroblast membranes, a traditional mechanism of action for long‐chain fatty acids. Alternatively, expression of free fatty acid receptor 4 (FFAR4), a G‐protein coupled receptor for long‐chain fatty acids, was detected in cardiac fibroblasts, suggesting an alternate mechanism of action. Our data suggest that EPA prevents interstitial fibrosis following TAC, potentially through FFAR4 in cardiac fibroblasts, which could have implications for the management of HFpEF.
Antiproliferative Effect of Kolaviron, a Biflavonoid Complex from the Seed of Garcinia Kola on Vascular Smooth Muscle Cells (VSMs) and A549 Cancer Cell Line
Authors: Oyagbemi Ademola , Omobowale Temidayo , Adedapo Adeolu , Oyekan Adebayo , Yakubu Momoh
Published: June 2021

Kolaviron (KV), a biflavonoid from Garcinia kola seeds has been reported to possess anti‐inflammatory, antioxidant, hepatoprotective, cardioprotective, nephroprotective and other arrays of chemopreventive capabilities. In this study, the antiproliferative potential of KV was investigated on Vascular Smooth Muscle Cells (VSMCs) in and A549 cancer cell vitro. Lipopolysaccharide (LPS) at 100µg/ml, 1 nm of Angiotensin II (10‐8) and 1 µM of Angiotensin II (10‐6) were used as mitogens to induce cell proliferation and 3‐(4, 5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide (MTT) assay was used to measure cell viability. Treatment of VSMCs with Angiotensin II (10‐8), 1 µM of Angiotensin II (10‐6) and LPS significantly enhanced proliferation of VSMCs. Also, treatment of VSMCs with LPS caused significant (p<0.05) increase in nitric oxide (NO) production. Furthermore, excessive NO production was significantly reduced following treatment with KV (25, 50 & 100 µg/ml) respectively. The mechanism of action of KV might be through its anti‐inflammatory and antioxidant capacities. Taken together, KV mitigates Angiotensin II and LPS induced cell proliferation, NO production, inhibited cyclic adenosine monophosphate (cAMP) and induced cytotoxicity in A549 cancer cell proliferation.
Garlic (Allium sativum) Modulates the Expression of the Receptor for Advanced Glycation End Product (RAGE) Isoforms in Renal Tissues of Streptozotocin‐Induced Diabetic Rats
Authors: Al‐Qattan Khaled , Mansour Mohamed , Thomson Martha , Ali Muslim
Published: June 2021

Background

The up‐regulation of the receptor for advanced glycation end products (RAGE) has been implicated as a major mediator in the development and progression of diabetic nephropathy.

Objective

The present study was designed to investigate the potential of garlic to modulate the level of expression of RAGE in renal tissues of diabetic rats.

Method

Three groups of rats were studied after 8 weeks following diabetes induction: normal, streptozotocin‐induced diabetic (control diabetic), and garlic‐treated diabetic rats.

Results

A polyclonal antibody of proven specificity to RAGE indicated in immunohistochemical assays that RAGE labeling was significantly increased in renal tissues of control diabetic rats compared to the normal group. The increased RAGE labeling involved mesangial cells in the majority of glomeruli, which also exhibited signs of mesangial expansion, mesangial nodule formation and glomerulosclerosis. In 2‐dimensional Western blots, RAGE expression in renal tissues was dominated by a heterogeneous group of charge variants, represented by 46‐50 kDa isoforms with an obvious shift towards more basic pIs compared to their counterparts in normal rats. Compared to control diabetic rats, RAGE labeling in the garlic‐treated diabetic group was significantly reduced throughout renal regions and was marked by the expression of 43‐50 kDa acidic charge variants comparable to those observed in normal rats.

Conclusion

The capacity of garlic to modulate diabetes‐induced up‐regulation of selective RAGE polymorphic variants may be implicated in attenuating the detrimental consequences of excessive RAGE signaling manifested by diabetes‐associated disorders.
Repeated Induction of Autophagy or Mitophagy Protects C2C12 Cells from Subsequent Apoptotic Insults
Authors: Bloemberg Darin , Quadrilatero Joe
Published: June 2021

Autophagy is a sub‐cellular degradation mechanism with critical roles in remodelling, defense, and protein/organelle quality control. Although recognized with pro‐death functions in certain instances, cells typically induce autophagy to mitigate stress as a means of cyto‐preservation. In fact, autophagy is suggested to contribute to the longevity benefits associated with various models of caloric restriction. However, limited knowledge exists regarding the importance and specific consequences of autophagy induction, particularly its potential ability to improve the cellular environment. In this study, we examined whether repeated bouts of autophagy or mitophagy could confer apoptotic‐resistance. Proliferating C2C12 cells were treated with HBSS or CCCP for 3 consecutive days to induce autophagy and mitophagy, respectively. Cells were allowed to recover for 24 hours, after which apoptotic stimuli were administered. Repeated HBSS and CCCP treatments each reduced (p<0.05) staurosporine‐induced activation of caspase‐3 and caspase‐9. Furthermore, in response to the Ca2+ ionophore A23187, HBSS‐ and CCCP‐treated cells maintained higher (p<0.05) calcein AM fluorescence and JC‐1 red:green fluorescence ratios compared to untreated cells. Therefore, mitochondria from conditioned cells preserved their membrane potential and were protected from permeability transition pore development after encountering Ca2+ stress. These results demonstrate that repeated induction of autophagy/mitophagy may have cyto‐protective effects, perhaps due to the development of more robust mitochondria.
Effect of Vitamin B12 and Nutrients on Adipogenesis‐adipogenic Markers in 3T3 Cells
Authors: Bellner Lars , Nichols Alexandra , Pandey Varunkumar , Vanella Luca , Gilliam Chelsea , Gupte Rahkee , Abraham Nader
Published: June 2021

Studies have shown that there is a deficiency of folate and vitamin B12 in obese individuals compared with normal BMI controls. Folic acid reduces oxidative stress while vitamin B12 protects against superoxide injuries in endothelial cells. We examined the hypothesis that folic acid and vitamin B12 on 3T3‐mediated adipogenesis by up‐regulating Wnt10b and heme oxygenase‐1 (HO‐1), thereby promoting the growth of small healthy adipocytes and reducing the large inflamed adipocytes. 3T3 cells were treated with either vehicle or one of four concentrations (0.1 to 100 μM) of either folic acid or vitamin B12 and effect on adipogenesis and adipogenic markers were studied. There was a decrease in adipocytes in a concentration‐dependent manner with an optimal level of 10 μM (p<0.05) in both folic acid and vitamin B12 treatment. Treatment of folic acid (10 μM) and vitamin B12 (10 μM) resulted in a decrease in the number of large, unhealthy adipocytes (p<0.05). The beneficial effects of folic acid and vitamin B12 were reversed when cells were treated with siRNA Wnt10b (p<0.05).These novel results demonstrate that increased levels of both folic acid and vitamin B12 attenuate adipogenesis via significant increases in Wnt10b and related proteins. Folic acid and vitamin B12 supplementation is deemed viable in the prevention and treatment of obesity and obesity‐associated diseases. Support NIH, HL‐55601, HL34300 (N.G.A.)
Double Inferior Venacava with Anomalous Hemiazygos Vein – A Case Report
Authors: Sakthivelavan Sumathilatha , Castellanos Bedia , Rich Sushama , Goldberg Robert , Quinteros Carlos , Baez Ramona
Published: June 2021

Objective

Duplication of the inferior venacava (IVC) is a rare variation that is caused by an alteration in the embryogenesis of the cardinal venous system. It is found in 0.3 to 3% of the population. There are various types of double IVC and this case is reported since it is unique among them.

Materials and methods: During dissection of an eighty seven year old female cadaver, presence of an IVC was noted on either side of the abdominal aorta. The course and the termination of both the IVCs were dissected. The azygos and the hemiazygos veins were traced into the thoracic cavity till their termination.

Results: In the posterior abdominal wall, the right IVC was found to be the continuation of the right common iliac vein and its course and termination was normal. On the left, the common iliac vein continued as the left IVC. Both the IVCs were connected by a venous bridge which coursed deep to the abdominal aorta. The left IVC passed through the medial arcuate ligament and entered the thoracic cavity as the hemiazygos vein. It was the only hemiazygos vein and drained into the azygos vein which was smaller than the hemiazygos vein.

Conclusion

The most common type of the double IVC is the termination of the left IVC in the left renal vein which drains into the right IVC. This case had a normal termination of the right IVC and continuation of the left IVC as the hemiazygos vein, which is a rarely reported variant. Double IVC is usually asymptomatic but if unnoticed, may lead to severe hemorrhage in the retroperitoneal surgeries. In the imaging studies, a thrombosed left IVC might mimic retroperitoneal lymphadenopathy. Adequate knowledge of this variant aids in the proper interpretation of radiological images and avoids intraoperative complications.
Characterization of the Activity and Biological Function of Human Protein Arginine Methyltransferase 9 (PRMT9)
Authors: Hadjikyriacou Andrea , Yang Yanzhong , Bedford Mark , Clarke Steven
Published: June 2021

Protein arginine methyltransferases (PRMTs) make up a family of enzymes that transfer methyl groups onto arginine residues and play important roles in DNA repair, splicing, transcriptional control and signaling. In mammals, a family of nine PRMTs are known. PRMT1,2,3,4,6, and 8 catalyze the formation of asymmetric dimethylarginine (ADMA) and w‐monomethylarginine (MMA), PRMT5 catalyzes symmetric dimethylarginine (SDMA) and MMA formation, and PRMT7 catalyzes only MMA formation. The last member of this family, PRMT9 (also designated PRMT10), has not previously been characterized. In humans, it is encoded on a gene on chromosome 4 in position 4q31. We have now expressed human PRMT9 both as a GST‐fusion protein in bacterial cells and as a GFP‐fusion protein in human HEK293 cells. For both expression systems, we have also prepared catalytic mutants to serve as controls for possible contamination of other types of PRMTs. Using an in vitro methylation assay, we show that both forms of PRMT9 catalyze the methylation of myelin basic protein and a GST‐fusion protein of the N‐terminal domain of human fibrillarin. Studies are in progress to characterize the type of activity and physiological substrates for PRMT9.
Sex‐Specific Metabolic Alterations in Offspring of Mice Exposed Prenatally to Particulate Matter (PM) from Petrodiesel (PD) Combustion
Authors: Fukagawa Naomi , Jetton Thomas , Peshavaria Mina , Bonney Elizabeth , Poynter Matthew , Holmen Britt
Published: June 2021

Epidemiological studies report that exposure of expectant mothers to outdoor air pollution correlates with an increased risk of obesity, insulin resistance (IR) and earlier onset of type 2 diabetes mellitus in her offspring. To test the hypothesis that prenatal exposure to PM from PD combustion will lead to alterations in weight (WT) gain and insulin sensitivity, pregnant C57BL/6J mice were exposed daily to <10% ethanol (control, CON) or to ~80 ug of PM suspended in <10% ethanol, mimicking daily ambient exposure, via oropharyngeal aspiration from gestational day 9‐17 and allowed to deliver. Mice were fed standard chow after weaning. Weekly body WT, food intake (FI), bi‐weekly fasting glucose levels were measured for 44 wks. While FI was similar between CON and treated groups, female offspring (F) of mothers exposed to PD gained less WT than CON (p<0.001) whereas male offspring (M) began gaining more WT than CON after ~24 wks of age. M (n=10) were also hyperinsulinemic and insulin resistant (homeostatic model assessment of IR, HOMA‐IR) compared to CON (n=3) at 24 wks (1.8±0.2 vs 0.9±0.09 ng/ml; HOMA‐IR 15±2 vs. 7±1, respectively; p<0.05, mean±SE) despite a similar degree of fasting glycemia (135±5 vs 137±5 mg/dl). In contrast, F remained normoglycemic with no difference in HOMA‐IR between CON (n=4) and PD‐exposed (n=9). These data suggest that prenatal exposure to air pollution particulates from PD combustion has different metabolic effects in M compared to F that portend differential risk for metabolic dysfunction and may influence future reproductive capacity. (Support: UVM Transportation Research Center Project #026904).
Effects of Green Tea and Coffee Polyphenols on Cardiometabolic Function in Women with Polycystic Ovary Syndrome
Authors: Tomatis Virginia , Wassell Sara , Venables Michelle , Walker Celia , Ray Sumantra , Siervo Mario , Griffin Julian , Bluck Les
Published: June 2021

Green tea (catechins, GTC) and coffee polyphenols (chlorogenic acids, CGA) have been associated with reduced diabetes and cardiovascular risk. However, detailed investigations in at risk populations are needed to corroborate their efficacy and understand the underlying mechanisms. This study aimed to evaluate the effects of a combined supplementation of GTC and CGA on insulin sensitivity (SI), nitric oxide (NO) production and cardiometabolic risk factors in women with polycystic ovary syndrome (PCOS), a condition characterized by insulin resistance, endothelial dysfunction and increased cardiometabolic risk. In a 16‐week, single‐blind, unilateral crossover trial, 12 PCOS women (mean ± SD: age, 28 ± 10 y; BMI, 35 ± 7 kg/m2; fasting insulin, 97 ± 52 pmol/L) underwent 8 weeks of placebo treatment followed by 8 weeks of active treatment (tablets providing 2093 mg GTC and 220 mg CGA/day). At baseline, post‐placebo and post‐active treatments participants undertook glucose tolerance and oral nitrate tests for isotopic assessment of SI and NO production and additional cardiometabolic risk factors were measured. The polyphenol intervention did not improve SI and NO production, but it was associated with reduced waist circumference (MD: 2.4 cm; P = 0.02), altered eicosanoid profile and non‐significant trends for reductions in diastolic blood pressure, total and LDL cholesterol. Combined intake of GTC and CGA may modulate eicosanoid pathways, thereby reducing inflammation and abdominal adiposity. These changes may prove cardioprotective in at risk populations, in longer term intervention studies.
[MRC UK]
The Effects of Folic Acid‐Fortified Food and Nutrition Education on Folate Nutritional Status and Behavioral Capability in Korean Women of Child‐Bearing Age
Authors: Hyun Taisun , Kim Min‐Ji , Kim Ji‐Hyun , Jang Young‐Ai , Kim Kyungmin , Lee Yoonna
Published: June 2021

The purpose of the study was to investigate the effects of folic acid‐fortified food and nutrition education on folate status and behavioral capability of Korean women of child‐bearing age. Instant rice fortified with 400 μg of folic acid per serving (one bowl) was developed. Seventy‐eight women aged between 19 and 35 who were at risk of folate deficiency were randomly assigned to 'non‐fortified food group', 'fortified food group', 'non‐fortified food and nutrition education group' and 'fortified food and nutrition education group'. All the subjects were asked to eat one bowl of fortified or non‐fortified food blindly once a day for 4 weeks. Nutrition education based on social cognitive theory was given to nutrition education group once a week. Questionnaire survey and hematological analysis were performed both at the baseline and at the end of the intervention. Serum and red blood cell folate concentrations increased significantly and serum homocysteine concentrations decreased significantly in the fortified food groups after the intervention. The proportions of folate deficiency decreased significantly in the fortified food groups. The variables of social cognitive theory including outcome expectations and behavioral capability score increased significantly in the nutrition education groups after the intervention. Our results suggest that folic acid‐fortified food is effective in improving folate nutritional status even after 4‐week supplementation, and nutrition education has a positive impact on the various factors related to behavior change.
Effects of Vitamin D Supplementation on Serum Adiponectin: a Metaregression Analysis of Clinical Trials
Authors: Minto Clara , Vecchio Maria Gabriella , Gregori Dario
Published: June 2021

Background

Vitamin D deficiency has been related to cardiometabolic risk factors. Recent studies suggest a potential association between low Vit D levels and adiponectin, a protein associated with obesity, T2DM and atherosclerosis. Present study evaluates, through a meta‐analysis, the association between serum vitamin D and adiponectin after vitamin supplementation.

Methods

Articles have been collected through PubMed using the search string “(vitamin D) AND adiponectin” limiting the results to “clinical trials” and “humans” and including papers published until December 2014. 6 articles have been chosen for the analysis. A meta‐regression model for plasmatic adiponectin has been estimated using Generalized Least Squares, accounting for within‐study correlation (assuming a Continuous Auto‐Regressive of first order structure) and weights proportional to study variance and study size. Effect of dose has been estimated as Cumulative Effect (Dose per day over the weeks of study duration).

Results

A total of 6 studies (175 patients) followed for an average time of 23 weeks were included in the meta‐analysis for serum adiponectin evaluation. A significant dose‐response relationship has been found between vitamin D3 supplementation and adiponectin levels.

Num. Studies
Effect
S.E.
p‐value

Dose on serum adiponectin (µg/mL)
6
0.0000039
0.00000047
<0.0001

Conclusions

Data suggest a potential increase of serum adiponectin after vitamin D intake. This result supports the assumption that vitamin D could play a role in the prevention of several diseases involving adiponectin variations. However, given the variability of sample characteristics and the variety of diagnostic tests, more research is needed to confirm present conclusion.
Examining the quality and student acceptability of school lunches in a Title 1, Alaska Native serving school district: findings from the Neqa Elicarvigmun study.
Authors: bersamin Andrea , Niles Kristine , Izumi Betty
Published: June 2021

The national school lunch program (NSLP) is particularly important in vulnerable communities with high rates of food insecurity. The nutritional quality and student acceptability of the lunch, however, affect and potentially limit the program's benefits to vulnerable students. Our objective was to examine the quality and student acceptability of the school lunch in a Title 1, Alaska Native serving school district participating in a Fish‐to‐School program. Lunch trays were photographed before and after students ate to estimate plate waste, which was used as a proxy for student acceptability. School lunch menus were entered into NDS‐R 2014 to estimate nutrient quality. On average, more than 50% of vegetables were thrown away. This average was significantly lower when the vegetables were included in a mixed dish (e.g. salmon soup). Less than 10% of the main entrées were thrown away. Entrees featuring salmon, a culturally important traditional food, were among the most popular. Menu modifications that are more closely aligned with traditional dietary patterns are needed to improve the acceptability of the school lunches, especially the vegetable component, in Alaska Native communities.
Focus Groups Support the Use of the Environment and Policy Assessment Observation (EPAO) for Obesity Prevention in Childcare
Authors: Shaw Ashley , Letiecq Bethany , Gallo Sina
Published: June 2021

Preschool age is a critical period for obesity prevention. Increasingly more children are being cared for outside the home; however, no national nutrition and physical activity guidelines exist for child care centers. The Environment and Policy Assessment Observation (EPAO) instrument was administered at two child development centers (CDC) located in Fairfax, VA. Sites were selected based on partial vs. non‐subsidized status. Focus groups were conducted with parents at each center to identify the factors which promote a healthy child care environment. CDC 1, serving a population of lower mean income level, scored significantly higher (69%) on the EPAO compared to CDC 2 (53%). Higher scores related to serving more meals family style, teachers eating with children, and encouraging new foods. The emergent themes of the focus groups reinforced these findings as CDC 1 parents, as compared to CDC 2 parents, felt better informed about nutritional policies and practices. Parents appreciated that the menu included wide variety, cultural exposure, and allergen/gluten free options. CDC 1 parents commented on the relationship that staff had with children stating that staff nurtured a calm and encouraging environment. CDC 1 parents felt family style meals promoted table manners, self‐sufficiency, and civility. The policies pertaining to birthdays and celebrations were also clearly articulated and favorable. This study suggests that parents support many of the policies and practices identified by the EPAO. National child care policies for nutrition and physical activity should integrate the use of assessment tools such as the EPAO in the prevention of childhood obesity.
Regulation of the NOGO‐Associated Enhancer
Authors: Pira Charmaine , Haro Endika , Oberg Kerby
Published: June 2021

NOGO‐A has been characterized as a neurite outgrowth inhibitor with the capacity to block axonal regeneration in the adult central nervous system (CNS); however, NOGO‐A is also expressed during embryonic development when neurite outgrowth is essential and regeneration is possible. We have identified a conserved noncoding region 88.5Kb upstream of the NOGO‐A/RTN4 gene. This sequence exhibits enhancer activity in neuroectoderm during neural tube development coincident with NOGO‐A expression. Specifically, enhancer activity localizes to the myelencephalon and the spinal cord in stage 10 chick embryos, with increased activity at the level of somites 1‐6. Several putative transcription factor binding sites are found in this NOGO‐associated enhancer (NAE) which may contribute to activity, including Pit‐1a, HNF3b and Sox3. Site‐directed mutagenesis of all three binding sites leads to a complete loss in enhancer activity. Isolated mutation of the Pit‐1a or HNF3b binding site results in no apparent change in enhancer activity, while isolated mutation of the Sox3 binding site results in decreased activity. These data suggest that regulation requires Sox3 in concert with either Pit‐1a or HNF3b binding sites for full activity. Additional studies are needed to confirm the functional role of this enhancer during CNS development.
Development and Evaluation of Highly Specific Monoclonal Antibodies against Human LGR5 for the Identification of Colon Cancer Stem Cells
Authors: Fu Wei , Yuan Kehu , Chen Caiwei , Wei Haitao , Lu Hsiangmin Emily , Ren Qi , Logis Evelin , Kelly Sean , He Wei‐wu , Ma Donghui
Published: June 2021

The cancer stem cells is a subpopulation of cancer cells which is responsible for cancer initiation, development and metastasis. Detection and identification of cancer stem cells have been considered as one of the most important objectives for clinical diagnostic and therapeutic purposes. Research evidences indicated that Leucine‐rich repeat containing G‐protein‐coupled receptor 5 (LGR5) is one of the biomarkers specifically expressed on colon cancer stem cells, which is activated by extracellular Wnt molecules to trigger canonical Wnt signaling and gene expression related to cancer development. Therefore, it is critical to develop an LGR5 antibody with great sensitivity and specificity to detect endogenous LGR5 expression in cancer stem cells.

In this research, several LGR5 mouse monoclonal antibodies were generated for multiple applications. These LGR5 antibodies are also highly sensitive for flow application on LGR5 stable expression cells and proved to be highly specific with our proprietary high density protein microarray chip assay. Furthermore, we also performed immunohistochemistry assays on colon cancer tissue blocks for diagnostic application. These newly generated LGR5 antibodies could provide new tools for earlier diagnosis and therapeutic guidance on colon cancers.
Alteration of Redox Homeostasis and Protein Expression of Constitutive Nitric Oxide Synthases Contributes to Erectile Dysfunction of Heart Failure Rats
Authors: Claudino Mario A , da Silva Fabio , Veiga Frederico , Gambero Alessandra , Priviero Fernanda , Antunes Edson
Published: June 2021

The association of REDOX homeostasis (RH) and NO/cGMP signaling to development of chronic heart failure (CHF) has been investigated, but, alterations in these mechanism in the erectile dysfunction (ED) present in CHF are not well known. This study examined whether ED in rats submitted to arteriovenous fistula (volume overload) is mediated by changes in RH and/or NO/cGMP signaling. After 4 wks, intracavernosal pressure (ICP) was used to evaluate the erectile function (in vivo). Nitrergic and neurogenic responses induced by electrical‐field stimulation were obtained of corpus cavernosum rats (CCR; in vitro). Protein expression of endothelial‐ and neuronal‐nitric oxide synthases (eNOS, nNOS) were evaluated in CCR, as well as, plasmatic activity of superoxide dismutase (SOD) and T‐BARS. CHF rats (CHFR) showed the reduction of ICP in all frequency (P<0.01) compared to sham rats (SR). Nitrergic relaxation in CCR of CHFR was decreased at higher frequencies (16‐32 Hz; P<0.05), but, neurogenic contraction was increased at 32 Hz (P<0.05), compared to SR. Protein expression of eNOS and nNOS in the CCR of CHFR where reduced in 37% and 44%, respectively (P<0.05). CHFR showed a decreased of plasmatic activity SOD and an increase of T‐BARS (P<0.05). In conclusion, CHFR exhibit a reduction of eNOS and nNOS expression resulting in an impaired nitrergic response and increased the neurogenic response contributing to reduction of ICP, as well as, promote alteration in the RH, thus, these elements may contribute to development of ED in the CHF. Financial Support: FAPESP/CNPq.
The Morphology of Male Gonads of the Neotenic Cave Salamander (Proteus anguinus)
Authors: Bizjak Mali Lilijana , Bulog Boris
Published: June 2021

Proteus anguinus is a cave‐dwelling urodele amphibian that is endemic to underground waters in Dinaric karst regions of Central Europe. It is an obligate neotene, with a very long life span and late sexual maturity. The reproductive period is a‐seasonal and lasts at least 30 years with each female laying eggs at intervals of 6 ‐ 12.5 years. The objective of this study was to describe the morphology of the testes of Proteus for better understanding of the reproductive strategy of this neotenic cave amphibian with the long‐term goal of establishing a facility for captive breeding. The testes used for this research were removed from our permanent collection of archived specimens. At least three morphological forms of testes are present independent of the size of the specimens, most likely representing phases of maturation of the gonads. The internal histological structure the testis is a simple cystic type of cyts enclosed in lobules with synchronous maturation of the germ cells within each cyst. The development of cysts inside a particular lobule is either synchronous or asynchronous. Spermatogenesis progresses in caudo‐cephalic direction of the testis as well as in proximal‐distal direction in relation to internal collecting ducts and appears to be independent of the seasons of the year. We can conclude that the reproductive peculiarities in Proteus probably reflect the fact that it lives in a relatively stable habitat with environmental conditions that are conducive to a‐seasonal reproductive cycles, similar to tropical salamanders. This research was supported by a grant from the Ministry of Higher Education, Science and Technology, Republic of Slovenia (P1–0184).
Exercise Training is More Effective Than The Ability of 17‐β‐Estradiol to Reverse Glucose Intolerance in Ovariectomized (OVX) Rats
Authors: MacDonald Tara , Ritchie Kerry , Davies Sarah , Hamilton Melissa , Cervone Daniel , Wright David , Dyck David
Published: June 2021

Introduction

Loss of estrogen in females increases the risk for insulin resistance. Hormone replacement therapy (HRT) is often prescribed to treat estrogen deficiency but has several detrimental side effects. Exercise is suggested as an HRT substitute since it improves insulin sensitivity, but the efficacy of exercise vs. HRT on glucose tolerance is unknown. The purpose of this study was to compare exercise training to 17‐β‐estradiol (E2) supplementation on restoring glucose tolerance (GT) in ovariectomized (OVX) rats.

Methods

30 OVX and 20 SHAM rats consumed a phytoestrogen free diet, ad libitum. Glucose tolerance tests (GTT) were performed at 10 weeks. Blood samples were drawn to assess serum insulin. Rats were randomly assigned to one of five 4 week treatment groups: SHAM sedentary (sed) or exercise (ex; 60 min, 5x/wk), OVX sed, ex or E2 (28 ug/kg/day). GTTs and blood sampling were repeated at 15 weeks. Glucose uptake in soleus and EDL muscle was assessed, and basal and maximal insulin‐stimulated EDL, soleus, adipose and liver samples were frozen for analysis.

Results

OVX rats were glucose intolerant at 10 weeks relative to SHAMs. After treatment, GT in OVX ex rats was entirely recovered, but only partially in OVX E2 rats. Insulin secretion at 15 min. during the 15 week GTT was significantly higher in OVX sed vs. OVX E2 or ex groups; however maximal insulin‐stimulated glucose uptake in soleus or EDL was unaffected.

Conclusions

Exercise restores GT in OVX rats more effectively than E2. Whole‐body improvements are not reflected by alterations in muscle maximal insulin response. This suggests adipose or liver as primary effectors of E2 or exercise treatment in OVX rats.

Supported by NSERC
Functional Expression of WNK Kinases and Insulin Signaling Effectors in Diabetic Skeletal Muscle
Authors: Kim Hanul , Kim Ji‐Hee , Hwang Kyu‐Hee , Park Kyu‐Sang , Cha Seung‐Kuy , Kong In Deok
Published: June 2021

WNK (with‐no‐lysine [K]) kinases (WNK1‐4) are serine/threonine protein kinases with an atypical placement of the catalytic lysine and tissue‐specific expression. WNK1, ubiquitously expressed including skeletal muscle, is known as a downstream effector of insulin signaling and WNKs have been implicated in protein trafficking highly correlated with insulin resistance or diabetes. However, expression pattern and functional role of WNKs in skeletal muscle and linkage between WNKs and insulin signaling effectors such as PKB/Akt and glucose transports in skeletal muscle with type II diabetes are ill‐defined. In the present study, WNK1 and WNK2, not WNK4 are expressed in C2C12. The expression levels of WNK1 and WNK2 significantly decreased in db/db mice, a type II diabetic model, compared to that of wild type mice. Phosphorylated Akt in skeletal muscle decreased in db/db mice supporting insulin resistance in skeletal muscle. Insulin stimulates Akt followed by WNK1 phosphorylation. Insulin stimulation of WNK1 is blunted by phosphoinositide‐3‐kinase inhibitors. Together, an expression of GLUT4, a downstream effector of Akt and/or WNK1, markedly reduced in db/db mice, whereas GLUT1 expression level is not altered suggesting that GLUT1 and GLUT4 are differently regulated in type II diabetic skeletal muscle. Therefore, these results may provide new insights for increased susceptibility to insulin resistance and diabetes in skeletal muscle.[This research was supported by Basic Science Research Program through the National Research Foundation of Korea funded by the Ministry of Education, Science and Technology (NRF‐2013R1A1A2060764)]
Extracellular Nucleic Acids in the Blood of Patients with Secondary Home Pneumonia with Chronic Obstructive Pulmonary Disease
Authors: Bakirova Ryszhan , Muravlyova Larissa , Molotov‐Luchanskiy Vilen
Published: June 2021

Insufficiently studied the role of extracellular nucleic acids. It is now established that the content of the circulating nucleic acids varies with diabetes, myocardial infarction, systemic lupus erythematous, rheumatoid arthritis, glomerulonephritis and hepatitis.

The aim of our study was to investigate the content of vkNK and acid precursors in plasma and adsorbed on erythrocytes of patients with secondary home pneumonia on a background of COPD.

Definition vkNK and RHA performed no later than one hour after blood collection. Statistical research methods included: payment median and mode, the mean square deviation. To determine the reliability of the indicators used F ‐ Fisher's exact test.

The first group consisted of 29 patients with secondary, home pneumonia, shared or with the defeat of two or three segments and the NAM II. The second group (n=20) included patients with no prior home pneumonia bronchitis or COPD background. The groups were matched by age and sex: the average age of the surveyed first was 54,8±16,5 years, the second ‐ 59,8±19,3 years, the control group ‐ 57,6±14,9 years. In all groups, dominated by men: the first ‐ 22 (75,8±7,9%), the second ‐ 17 (85±7,9%), in the control group ‐ 19 (76±8,5%).

In 8 (27,6±8,3%) patients of the group identified COPD stage II, and 21 (72,4±8,3%) ‐ III stage. The mean value of the duration of COPD group was 10,4±8,7 years. The frequency of exacerbations of COPD equal 2,2±1,2 years.

In the red blood cells of patients with secondary home pneumonia against a sharp increase in COPD CRF adsorbed on erythrocytes. Thus, the totality of our data allows us to offer the extracellular nucleic acids are considered as a separate component of the molecular patterns associated with damage with pneumonia
Alteration of the PKA-CREB cascade in the mPFC accompanying prepulse inhibition deficits: evidence from adolescent social isolation and chronic SKF38393 injection during early adolescence
Authors: Du Wei , Li Man , Zhou Hao , Shao Feng , Wang Weiwen
Published: June 2021

Prepulse inhibition (PPI) refers to the inhibition of the startle reflex that occurs when the startling stimulus is preceded by a weak prestimulus. Altered adolescent mPFC circuitry induced by early-life adversity might be a key source of PPI deficits. The current study focused on variations in the cyclic AMP (cAMP)/protein kinase A (PKA)-cAMP-response element-binding protein (CREB) pathway in the medial prefrontal cortex (mPFC). We found a negative relationship between PPI and the PKA-CREB cascade during adolescence by employing both developmental and pharmacologic manipulations. Experiment 1, with the early adolescent social isolation model [postnatal days (PNDs), 21–34), displayed a disrupted PPI at PND 35 and significantly altered PKA, phosphorylated CREB (p-CREB) and the ratio of p-CREB to CREB. In particular, the level of p-CREB was negatively related to PPI performance. In Experiment 2, SKF38393, a well-characterized activator of adenylate cyclase and cAMP/PKA, was chronically injected during early adolescence (PNDs 28–34). We sought to mimic potential biochemical changes, particularly PKA activation, which is possibly altered by adolescent social isolation, and to determine if PPI was disrupted, similar to the disruption associated with adolescent social isolation. On PND 35, PPI deficits were detected, as well as increased PKA, marginally increased CREB and no change occurred in p-CREB or the ratio of p-CREB to CREB. In particular, PKA activity was negatively related to PPI performance. Although these results are limited in suggesting a causal link between PPI deficits and PKA-CREB signaling, they may help to elucidate the role played by PKA-CREB in the mPFC in regulating PPI.