bluuebunny/crossref_metadata_2025_split huggingface dataset:
Role of Sororin in sister chromatid cohesion and separation in vertebrates
Authors: Zhang Nenggang , Panigrahi Anil K , Jiang Yunyun , Rao Pulivarthi H , Pati Debananda 
Published: June 2021

Sororin, a cohesin associated protein, is conserved in vertebrates but has no apparent homolog in lower eukaryotes. It is expressed in S phase and degraded in early G1 phase. Co‐expression analysis indicates that Sororin gene, CdcA5, is co‐regulated with known cell cycle genes, such as Cdk1, Cyclin B, Bub1, Cdc20 etc. High‐throughput protein phosphorylation analysis indicates that Sororin is phosphorylated during CDK‐driven phase of cell cycle. Sororin interacts with cohesin complex and is required for sister chromatid cohesion. Here we report that Sororin mediates the maintenance and removal of sister chromatid cohesion. Co‐immunoprecipitation and in vitro binding assay show that Sororin interacts with cohesin complex via cohesin subunit SA1/2. Sororin also interacts with Plk1 via a PBD binding motif. Mutation on PBD binding motif of Sororin or PBD of Plk1 inhibits the interaction of Sororin and Plk1. Since phosphorylation of SA2 by Plk1 is required for the removal of sister chromatid cohesion on chromosomal arms, it is possible that Sororin acts as a docking protein to recruit Plk1 to phosphorylate SA2 and facilitates the dissolution of cohesin on chromosomal arms.(Supported by National Cancer Institute grant 1 R01 CA109330)
Molecular mechanism of RBP4 on the basis of insulin signaling pathway in primary hepatocytes
Authors: Namkung Jun , Kim Moon Young , Woo Mi Kyoung , Sohn Joon Hyung , An Sung Wan , Yeh Byung‐Il 
Published: June 2021

Metabolic syndrome is a cluster of symptoms closely associated with diabetes, obesity, and cardiovascular disease such as hypertension. Thought the prevalence is being increased the mechanism is not known so far. In this study, we tried to confirm the molecular mechanism of RBP4 which is regarded one of the critical factors in the pathogenesis of metabolic syndrome.Recombinant mouse RBP4 was treated to the primary hepatocytes prepared from the ICR mouse and cDNA microarray was performed. We focused insulin signaling pathway and mRNA expressions were measured real‐time PCR. The results are: 1) RBP4 has an undesirable effect for metabolic syndrome by increase of gluconeogenesis and decrease of glycolysis. 2) The increase of gluconeogenesis is due to decreased expression of Akt. 3) The effects of several factors which influences on PIP3 expression, which plays a role as controller at the upper level than PKCα and Akt in the insulin signaling pathway, are not constant by RBP4.In summary, RBP4 increase gluconeogenesis mediated by Akt and decrease glycolysis in insulin signaling pathway. To get the clue to solve the metabolic syndrome the researches to confirm the phosphoryation of the factors and protein expression analysis are needed.
Effects of 2‐Chloroethyl ethyl sulfide(CEES) on Guinea Pig Brain Morphology
Authors: Gadsden‐Gray Jessica , Mukherjee Shyamali , Ogunkua Olugbemiga , Das Salil K. 
Published: June 2021

Mustard gas is a chemical warfare agent used in World Wars I and II, the Gulf War, and on civilian occupants of the Iranian village of Halabja. Inhalation of sulfur mustard is known to cause hemorrhagic inflammation of the tracheobranchial tree in addition to severe pulmonary complications such as adult respiratory distress syndrome (ARDS). Studies of the mustard gas analog 2‐chloroethyl ethyl sulfide (CEES) using the guinea pig model system have indicated that its inhalation not only has the ability to cause significant lung damage but also to cross the blood‐brain barrier as evidenced by down‐regulation of the pro‐opiomelanocortin (POMC) gene. Down‐regulation of the POMC gene also suggests abnormal brain function. CEES is known to decrease antioxidant defense systems and increase lipid peroxidation in the brain, which may occur by microglial activation. Increasing microglial number and migration in the brain can cause inflammatory and NADPH‐derived stress. Activated microglia are also known to play an important role in the development of neurodegenerative diseases such as Parkinson's disease and amylotropic lateral sclerosis(ALS). The purpose of this study was to determine if intratracheal CEES infusion causes abnormality in brain morphology in guinea pigs. Our results show that intratracheal CEES exposure causes changes in brain morphology that result in dose dependent increases in blood cell aggregation and microglial cell number, microglial activation, and brain inflammation. These changes may be mediated by induction of oxidative stress.This project was supported by grants from the Department of the Army (W81XWH‐06‐2‐0044) and NIH (5T32HL007735‐12).
Robotic high throughput fatty acid analysis of umbilical cord serum
Authors: Lin Yuhong , Hibbeln Joseph R , Zhou Weiyin , Loewke James , Brown James A , Lands William EM , Wells Ellen M , Goldman Lynn R , Salem Norman 
Published: June 2021

Several epidemiology studies show that inadequate dietary intakes of highly unsaturated omega‐3 fatty acids are adversely related to child IQ and aggressive behaviors. As a long term goal, our laboratory is developing several techniques to facilitate large scale quantification of plasma or serum fatty acids. These include the automation of chemical procedures for transmethylation and extraction using a robotic system, analytical analysis using fast gas‐liquid chromatography and automated peak identification and calculation of fatty acid content. The system has been validated as follows: (1) Fatty acid recovery was 97% on average at 3 different concentrations. (2) The CV for the 24 fatty acids over 47 replicates was 3.26±1.2% for concentrations and 2.44±1.3% for fatty acid percentages. A Microsoft Visual Basic program was written to automate data calculation and peak identification. The complete procedure for robotic transmethylation of up to 72 samples currently requires about 6 hours. A total of 300 umbilical cord serum samples were assayed for fatty acids profiles using this system as an initial application of the high throughput methodology.This project was partially funded by the DICBR, NIAAA, NIH.
Repressive effect of polyunsaturated fatty acids on serine dehydratase expression in rat hepatocytes
Authors: Contreras Alejandra , Ortiz Victor , Torres Nimbe , Tovar Armando R 
Published: June 2021

Peroxisome proliferator‐activated receptor alpha (PPARα) is activated by ligands such as polyunsaturated fatty acids (PUFAs) and regulates expression of enzymes involved in fatty acid oxidation. Recently, it was found that PPARα also regulates the metabolism of amino acids in liver, however, the knowledge of molecular mechanisms of amino acid regulation in liver is limited. To assess the effect of natural (linoleic acid, LA) and synthetic PPARα ligands (Wy14643) on the expression of the amino acid degrading enzyme serine dehydratase (SHD) and the enzyme involved in fatty acid oxidation carnitine palmitoyl transferase‐1 (CPT‐1), rat hepatocytes were incubated with the agonist Wy14643, palmitic acid (saturated fatty acid) or linoleic acid (PUFA), and gene expression was measured. Addition of forskolin to the medium increased SDH gene expression as expected but did not altered CPT‐1 expression. However, addition of linoleic acid and palmitic acid to the medium increased CPT‐1 by 100% and 40%, but repressed SDH gene expression by 53% and 23% respectively. These data indicated that the insaturated linoleic acid reduced amino acid catabolism and stimulated fatty acid oxidation through PPARα in a higher extent than the saturated palmitic acid.Supported by CONACYT and INCMNSZ.
Tropomyosin Phosphorylation in Healthy and Diseased Hearts
Authors: Sheikh Hajer N , Wieczorek David F 
Published: June 2021

Tropomyosin (Tm) is an alpha‐helical coiled coil dimer involved in regulating striated muscle contraction. Although sarcomeric Tm is known to be phosphorylated at the penultimate amino acid Ser‐283, little is known regarding its phosphorylation levels in healthy and cardiac diseased states. The objective of this study was to study Tm phosphorylation levels with respect to age in wild type mice and in mice with familial hypertrophic cardiomyopathy (FHC). Phosphorylation levels were determined by western blotting of 2‐D gel electrophoresis of myofibrillar protein preparations. Results show that cardiac Tm phosphorylation is high during the perinatal period and decreases throughout adulthood in wild type mice. We also found that walls of the different cardiac chambers exhibit differential Tm phosphorylation.Two transgenic mouse models harboring FHC mutations in alpha‐Tm (Glu180Gly, and Asp175Asn) were studied to determine the effect of this disease on cardiac Tm phosphorylation. Results show that Tm phosphorylation is altered in both FHC models. Unlike in wild type mice, Tm phosphorylation levels do not decrease after birth, but rather remain constant. Depending on the role of Tm phosphorylation, modulation of phosphorylation levels in FHC patients may be potentially of therapeutic value.This project was funded by NHLBI HL81680 grant to DFW. HNS was supported by a fellowship from Dubai Medical College.
Characterization of methyl group metabolism in the non‐obese diabetic (NOD) mouse
Authors: Neuman Joshua Charles , Nieman Kristin M , Schalinske Kevin L 
Published: June 2021

Perturbed methyl group metabolism leads to elevated circulating levels of homocysteine, an independent risk factor for cardiovascular disease, as well as DNA hypomethylation. A streptozotocin‐induced type 1 diabetic rat model showed disrupted methyl group metabolism, characterized by increased levels of glycine N‐methyltransferase (GNMT), DNA hypomethylation, and hypohomocysteinemia. The present study examined similar measures of methyl group metabolism in the genetic non‐obese diabetic (NOD) mouse model. At 24 wk of age, NOD and ICR (control) mice were sacrificed and blood, liver, and kidney tissue were collected. NOD mice displayed 59% (P < 0.05) greater GNMT activity compared to the ICR mice. A correlation between both hepatic GNMT activity (r = 0.81, P < 0.002) and DNA hypomethylation (r = 0.59, P ≤ 0.05) with plasma glucose concentrations was also observed. Plasma homocysteine levels in the blood decreased by 53% in the NOD group (P < 0.005) compared to the controls. However, phosphatidylethanolamine N‐methyltransferase (PEMT) was down‐regulated in NOD mice, compared to its increase with streptozotocin treatment. This indicates that a genetic and chemically induced model for type 1 diabetes exhibit similar, but not entirely consistent, findings with respect to disrupted methyl group metabolism.Grant Funding Source: NIH HD057315
tRNA (Ser)Sec coexpression affects SECp43 levels in HEK 293 cells
Authors: Gilman Christy Leinaala , Seale Lucia A , Seyedali Ali , Berry Marla J 
Published: June 2021

Selenium is present in the active site(s) of selenoproteins in the form of selenocysteine (sec), the 21st amino acid. Sec is incorporated cotranslationally into proteins through recoding of the UGA codon. Key factors in this process include a unique tRNA for Sec (Sec‐tRNA), SECIS elements, SBP2 and EFsec. Additional factors include SLA/LP, SPS1 and SECp43. Previous studies in our laboratory have found that cotransfection of these three factors resulted in coprecipitation in nuclear and cytosolic fractions, indicating the formation of complexes. In GST pull downs, SECp43 and SPS1 coimmunoprecipitate in the cytosol in the absence but not in the presence of Sec‐tRNA. SECp43 is present in the nuclear fraction in the absence of other factors, in either the absence or presence of Sec‐tRNA. SECp43 pull‐down of SPS1 or vice versa is not seen when SLA/LP is added, suggesting mutually exclusive interactions. SECp43 in the presence of SLA/LP pellets with V5 beads independent of V5 tagged protein, suggesting it may be part of a large complex. This is not seen when SPS1 is also present, further supporting mutually exclusive interactions. We are currently following up on these findings to elucidate the mechanism by which Sec‐tRNA affects SECp43 levels and to understand the interactions of these factors.Supported by NIH grant DK‐47320 to MJB.
The mismatch repair protein hMLH1 regulates selenium‐induced DNA damage response in colorectal cancer cells
Authors: Qi Yongmei , Schoene Norberta W , Cheng Wen‐Hsing 
Published: June 2021

Epidemiological studies indicate that selenium supplementation in humans suppresses risk of colorectal and other cancers. The majority of colorectal cancer is characterized by a defective DNA mismatch repair (MMR) system. The hMLH1‐hPMS2 heterodimer plays a critical role at the early stage of MMR repair. Here, we employed HCH116 colorectal cancer cells with intrinsic hMLH1 deficiency and the cell complemented with hMLH1, and treated them with sodium selenite (2–10 μM), methane seleninic acid (2–10 μM) and methyl‐seleno‐L‐cysteine (50–300 μM). We showed that complementation of hMLH1 sensitizes HCT116 cells to cell death after treatment with the selenium compounds. Interestingly, cellular exposure to organic forms of selenium compounds resulted in increased hMLH1 protein expression and hMLH1‐hPMS2 association. Furthermore, hMLH1 is involved in proper S and G2/M phase checkpoint responses and DNA‐PKcs phosphorylation at Thr‐2647, an early event of DNA damage response, after cellular exposure to selenite. Our results suggest that DNA damage checkpoint response, a barrier of tumorigenesis, is deficient in HCT116 cells after selenium treatment. Consequently, a lack of functional hMLH1‐hPMS2 complex may predispose colorectal cancer cells to genomic instability.Grant Funding Source: University of Maryland
ATM is upstream of DNA‐PKcs in the activation of DNA damage response by sodium selenite
Authors: Rocourt Caroline , Cheng Wen‐Hsing 
Published: June 2021

The role of selenium in mitigating tumorigenesis is not fully understood. Selenium in supranutritional doses can induce ROS, which indirectly causes DNA breaks. Previous reports indicate an interplay between Ataxia‐telangiectasia mutated (ATM) and DNA‐dependent protein kinase catalytic subunit (DNA‐PKcs) in the early stages of DNA damage response. We recently found that ATM is required for a selenium‐induced senescence response in non‐cancerous cells at an early stage. Thus, we hypothesize that the selenium‐induced DNA damage response can modify this ATM‐DNA‐PKcs cross‐talk. Phospho‐specific antibodies against ATM and DNA‐PKcs were used to follow the phosphorylation events after selenium treatment in MRC‐5 normal fibroblasts and ATM shRNA and control shRNA U‐2 OS osteosarcoma cells. Results from immunofluorescence analysis showed that selenium treatment induces phosphorylation of DNA‐PKcs at T2647, an ATM‐dependent phosphorylation site, and phosphorylation of DNA‐PKcs at S2056, an autocatalytic site of DNA‐PKcs, in non‐cancerous MRC‐5 cells but not in U‐2 OS cancer cells. Further studies in MRC‐5 cells with an ATM kinase inhibitor, KU55933, showed attenuation of the selenium‐induced DNA‐PKcs phosphorylation at both foci, whereas pre‐treatment with a DNA‐PKcs kinase inhibitor, NU7026, does not prevent ATM phosphorylation at S1981, an event leading to ATM pathway activation. The selenium‐induced DNA‐PKcs phosphorylation at S2056 is decreased in S‐phase cells. Taken together, our results provide the first evidence of ATM and DNA‐PKcs activation as an early tumorigenesis barrier in response to selenium exposure in non‐cancerous cells, and places ATM upstream of DNA‐PKcs in the selenium‐induced DNA damage response process.Grant Funding Source: University of Maryland
Possible rise in Ca:Mg ratio from food intake in U.S.A. adults
Authors: Rosanoff Andrea 
Published: June 2021

A growing body of work suggests an importance for Ca:Mg ratio. We calculated the change of Ca:Mg intake from foods over time in the United States of America.MethodU.S. Department of Agriculture Food Surveys from 1977, 1985, 1994, 2001–2, 2003–4, and 2005–6 provided mean Ca, Mg and Kcal intake per day in adult age‐gender groups. (USDA Surveys before 1977 do not include Mg; Data for adults age 50+ were not reported until 1994; Defined age/gender groups changed slightly for age from survey to survey.) Survey means were used to calculate % changes in Ca, Mg, Kcal, and Ca:Mg intakes from food for 1977 to 2006 (1994 – 2006 for age 50+ yrs).ResultsMean Ca daily intake rose in each age/gender group: a low of 21% for elderly males and a high of 77% for adult females. Mean Mg daily intakes rose in each adult age‐gender group, but to a lesser extent than Ca, from 3% for elderly males and a high of 28% in adult females. Caloric intakes rose in each age/gender group comparable to the rise in Mg rather than to the rise in Ca: 7% for elderly males and 26% for adult females.ConclusionLarge rises in Ca intakes between 1977 and 2005–6 result in an apparent rising Ca:Mg from food, observed from 1977 to 2005–6 for each adult age‐gender group; 20 – 38% rise for adult women and 18 – 32% rise for adult men. Supported by Center for Magnesium Education & Research.
The vitamin A equivalence of β‐carotene in β‐carotene‐biofortified cassava ingested by women
Authors: Liu Wenhong , Zhou Yang , Sanchez Teresa , Ceballos Hernan , White Wendy S. 
Published: June 2021

β‐Carotene‐biofortified cassava is being developed through plant breeding as a sustainable agronomic approach to alleviate vitamin A deficiency. Our objective was to quantify the vitamin A equivalence of the β‐carotene in biofortified cassava based on ingestion of a single realistic serving of cassava porridge. Women (n = 8) each consumed in random order 3 cassava porridges (220 g each) containing 40 g cassava flour: 1) biofortified cassava porridge (1097.5 μg β‐carotene); 2) white cassava porridge with β‐carotene reference dose (537.6 μg added β‐carotene); 3) white cassava porridge with vitamin A reference dose (285.6 μg added retinol). Blood samples were collected over 9 h. Retinyl palmitate was analyzed in triacylglycerol‐rich lipoprotein fractions by HPLC with electrochemical detection. Areas under the curve (AUC) for retinyl palmitate (nmol·h) were 213.0 ± 59.7, 141.5 ± 46.3, and 159.0 ± 81. 6 after ingestion of biofortified cassava porridge, white cassava porridge with β‐carotene reference dose, and white cassava porridge with vitamin A reference dose, respectively. The vitamin A equivalence of the β‐carotene in β‐carotene‐biofortified cassava was 2.80 ± 1.77 to 1 (by wt); the vitamin A equivalence of the β‐carotene reference dose (2.11 ± 0.81 to 1) approximated that established by the U.S. Institute of Medicine. Biofortified cassava is a highly bioavailable source of vitamin A. Supported by HarvestPlus.Grant Funding Source: HarvestPlus
Urinary excretion of chlorogenic acids and metabolites in humans after green mate (I. paraguariensis ) consumption.
Authors: Marques Viviane Xavier , Farah Adriana 
Published: June 2021

The consumption of green mate (I. paraguariensis) has been associated with health benefits, attributed mainly to its high chlorogenic acids (CGA) contents. It is known that CGA from coffee matrix are bioavailable to humans, but their bioavailability when in mate matrix has not been investigated. The aim of this study was to measure the urinary excretion of CGA and metabolites after green mate consumption. On separate days, subjects (n=7) consumed capsules of green mate (6.2μmol of CGA/Kg weight) or placebo. Urine samples were collected at baseline and for 24h after each treatment. CGA and metabolites were measured in urine by HPLC/DAD. Six CGA (3 caffeoylquinic and 3 dicaffeoylquinic acids) and 14 metabolites (caffeic, vanillic, ferulic, isoferulic, p‐coumaric, gallic, p‐hydroxybenzoic, dihydrocaffeic, syringic, sinapic, 3,4‐dihidroxyphenilacetic, 3‐(4‐hydroxyphenil)propionic, trans‐3‐hydroxycinnamic and benzoic acids) were quantified in the urine samples. The total urinary excretion after green mate (516.6μmol) consumption was higher (p<0.05) than placebo (374.7μmol). On average, after green mate consumption, 55% of total hydroxycinnamic acid and quinic acid moieties’ derivatives was recovered in urine. However, the existence of phenolic acids in urine after the placebo treatment should be considered. This study indicates that CGA from green mate are absorbed and metabolized in humans.Financial support: CNPq and FAPERJ, Brazil.
A simple and rapid method for induction of experimental allergy in mice
Authors: Christopher Lawrance , Kuvibidila Solo 
Published: June 2021

Ovalbumin‐sensitized mice are widely used animal models for allergy and asthma. The typical protocol requires a month to induce allergy and hence time consuming. Moreover, this protocol requires inhalation apparatus for mice therefore limiting this procedure to highly equipped laboratories. In this study we developed a short term protocol (15 days) for inducing allergy and used nasal instillation without using the inhalation apparatus. Fourteen weeks old C57BL/6 female mice fed the AIN76 diet for 2.5 months were randomly divided into two groups of 16 mice each. Experimental mice received two intraperitoneal injections of ovalbumin (16 μg ovalbumin + 2 mg of aluminum oxide dissolved in 0.4 ml PBS) at day 0 and day 6 whereas control mice received 0.4 ml of PBS. On the day 12 and 13 mice received nasal instillation of either 0.1 ml of PBS for control mice or 0.1 ml of PBS+ 200 μg of ovalbumin for treatment mice. Mice were sacrificed on day 14 of the protocol, and plasma was used for IgE assay by ELISA. There was no significant difference in mean body weight at time 0. There was a 4‐fold increase in plasma total Ig E in ovalbumin‐treated (127.524 ng/ml) versus control (31.50 ng/ml) (p <0.05). Our improved method not only reduced the time period but also the requirement of inhalation apparatus for mice. Hence this method is rapid and simple to induce experimental allergy in mice.Grant Funding Source: Mushroom Council & Oklahoma State University
Nutritional support for piglets simulating nutrition support in the NICU
Authors: Smith Kristin , Borum Peggy R. 
Published: June 2021

BackgroundPiglets in the Piglet Neonatal Intensive Care Unit (PNICU) can serve as models for neonates in the Neonatal Intensive Care Unit (NICU) and can be used to address nutrition questions requiring techniques too invasive to conduct in neonates. Nutrition support must meet the metabolic needs of the piglets and simulate NICU nutrition support. Defined piglet formulas are needed to allow comparison of control to experimental groups.ObjectiveTo create defined formulas for PNICU nutrition support simulating NICU nutrition support that achieves optimal health and growth of term and preterm piglets.MethodsNutrient data from pig production studies, piglet model investigations, and human preterm and full term nutrition trials were used as reference data and formulas were defined using commercially available constituents.ResultsThe following enteral formula for a term piglet and parenteral formula for a preterm piglet were developed.

Nutrient
Term Enteral
Preterm Parenteral

Protein
7.25 g/kg*d
8 g/kg*d

Carbohydrate
21.96 g/kg*d
12 g/kg*d

Lipid
15.58 g/kg*d
4.5 g/kg*d

Calcium
0.49 g/kg*d
0.1 g/kg*d

Phosphorus
0.47 g/kg*d
0.059 g/kg*d

Ca:P
1.04:1
1.7:1

Total Calories
257.02 kcal/kg*d
141 kcal/kg*d

ConclusionsThe establishment of these piglet formulas is an important step in refining translational research using term and preterm piglet models for term and preterm human neonates.