bluuebunny/crossref_metadata_2025 huggingface dataset:
Alpha‐keto acid metabolites of organoselenium compounds inhibit HDAC activity
Authors: Nian Hui , Dashwood Roderick H
Published: June 2021

HDAC inhibitors are gaining interest as cancer therapeutic agents. We hypothesized that natural organoselenium compound Se‐methylselenocysteine (MSC) might be metabolized to an HDAC inhibitor. In a cell‐free assay, glutamine transaminase K (GTK) converted MSC to methylselenopyruvate (MSP). Molecular modeling supported the interaction of MSP with zinc in the HDAC pocket. In an in vitro HDAC activity assay, MSP was a competitive inhibitor. In various cancer cell lines, MSP increased acetylated histone levels, and induced cell cycle arrest and/or apoptosis. P21WAF1 was increased at both the mRNA and protein level by MSP, and there was enhanced P21WAF1 promoter activity. This work provides a new paradigm by which organoselenium compounds might protect against prostate, colon, skin and other cancers. Thus, in addition to targeting redox‐sensitive signaling molecules, alpha‐keto acid metabolites of organoselenium compounds might alter HDAC activity and histone acetylation status.
Imidazole inhibits B16 melanoma cell migration via degradation of beta‐catenin
Authors: Jeong Yun Mi , Li Hai Lan , Kim Su Yeon , Yun Hye‐Young , Baek Kwang Jin , Kwon Nyoun Soo , Kim Dong‐Seok
Published: June 2021

We investigated whether imidazole inhibits the migration of B16 murine melanoma cells. First of all, we found that imidazole did not show any cytotoxic effect at less than 100 μM in B16 cells and HaCaT cells (normal human keratinocytes). In addition, imidazole had no influence on B16 cell proliferation at the same concentration range. To explore the effects of imidazole on B16 cell migration, we treated imidazole to B16 cells at various concentrations and measured the migration using Scratch Wound Migration Assay. In 6 h, imidazole significantly inhibited B16 cell migration in a concentration‐dependent manner. It is known that migration involves phosphorylation of two signaling pathways; Akt‐ and GSK3β‐activated protein kinase pathway. In 1 h, Western blotting results showed the inhibition of both Akt‐ and GSK3β pathway in imidazole‐treated B16 cells. Moreover, imidazole also led to the degradation of β‐cateninin via GSK3β pathway. Further results showed that MG132, a proteasomal inhibitor, abrogated cell migration by imidazole in B16 cells. Thus, we demonstrate that imidazole inhibits cell migration in B16 cells by β‐catenin degradation. Moreover, we suggest that imidazole may serve as a chemo‐preventive candidate agent for melanoma therapy.
Enzyme involved in degradation of isonitrile with a nitrogen‐carbon triple bond
Authors: KOBAYASHI MICHIHIKO , SATO HIROYOSHI , HASHIMOTO YOSHITERU
Published: June 2021

We have studied microbial metabolism of nitrile [R‐CN] and its isomer, isonitrile [R‐NC], both of which contain a C‐N triple bond. Although isonitriles are widely distributed in nature, information on their biological metabolism is quite limited. We obtained isonitrile‐degradable microorganisms, clarified the degradation pathway and discovered two new enzymes. Isonitrile hydratase (InhA) from Pseudomonas putida catalyzes hydration of isonitrile to N‐substituted formamide [R‐NH‐CH(=O)]. N‐substituted formamide deformylase from Arthrobacter pascens converts N‐substituted formamide to amine and formate. They have been approved as new enzymes by NC‐IUBMB. The upstream metabolism of isonitriles remains unknown in A. pascens. Therefore, we here investigated it and discovered another new enzyme involved in isonitrile degradation. At first, we found that the highest activity of isonitrile hydratase was obtained by the addition of N‐benzylformamide to the Arthrobacter cells. The isonitrile hydratase designated InhB was purified from the cell‐free extracts, and found to act on isonitriles to yield the corresponding N‐substituted formamide. InhB exhibited maximum activity at pH 7.3–7.7. Western blot analysis of the purified InhB using the anti‐InhA polyclonal antibody revealed that the primary structure of InhB did not show any similarity to that of InhA.
Control of intramuscular fat accumulation in porcine muscle by dietary lysine levels
Authors: Katsumata Masaya , Matsumoto Mitsuhito , Kawakami Shin‐ichi , Ishida Aiko , Kyoya Takahito , Nakashima Kazuki , Kaji Yuji , Takada Ryozo
Published: June 2021

Pigs of six weeks of age were given a control or low lysine (LL) diet for three weeks. GLUT4 mRNA expression was up‐regulated by the LL diet in l. dorsi and rhomboideus muscles. Activities of citrate synthase were higher in those two muscles of the LL group suggesting that oxidative function of the muscles was enhanced by dietary low lysine. Thus, we determined proportion of oxidative myofiber. Indeed, proportion of oxidative fiber was higher in the LL group than in the control group. As higher triglyceride and/or intramuscular fat (IMF) content are associated with higher proportion of oxidative myofiber, we hypothesized that accumulation of IMF might be promoted by dietary low lysine. To test this hypothesis, a low lysine diet was given to finishing pigs for two months. IMF contents in l. dorsi of the pigs given a low lysine diet were twice higher than those of the control group. We also elucidated effects of dietary lysine levels on expression of adipogenesis related genes in l. dorsi and rhomboideus muscles of growing pigs; abundances of mRNA of PPARγ tended to be higher in the LL group whereas dietary lysine levels did not affect that of C/EBPα. We infer that differentiation of preadipocytes to adipocytes is not involved in promoted accumulation of IMF due to dietary low lysine in porcine muscle.
Characterization of charge‐variant bioconjugates derived from chemical labeling of proteins
Authors: Burgos Glamarie , Rae Tracey , Bonn Ryan , Fishpaugh Jeff , Munoz Joseph , Pope Mark , Bax Brian
Published: June 2021

Purified antibodies and other target‐specific recognition proteins are commonly used reagents in a wide range of biochemical analyses from basic research to the clinical laboratory. Characterization of charge states for these purified protein reagents is often critical to indication of solution‐phase stability and the potential for non‐specific interactions with solid‐phase materials and other proteins. Conjugation, or chemical labeling, with various small molecules adds further complexity to the inherent charge‐variation from isoforms of a single purified protein. While these added labels typically cause minimal change in the overall size of the protein molecule and infrequently inhibit the native protein function, they can cause a significant change in effective protein charge. The experiments described here examine the effect of an amine‐reactive protein labeling process on the charge state distribution of resulting bioconjugates. Various analytical methods were applied to resolve and characterize the changes in protein charge state distribution as a function of the labeling load (label‐to‐protein ratio, or incorporation ratio (IR)) and as a function of the type of label used. A distinction between surface charge effects and net protein charge effects is addressed.
Systems biology approaches to characterize exposures to infectious agents
Authors: Hammamieh Rasha , Jett Marti
Published: June 2021

There is an urgent need to assess exposure and infection of individuals by biological threat agents (BTA) well in advance of onset of illness or at various stages post‐exposure.

The rapid development of genomics, proteomics and high‐throughput computing approaches to study diseases is now allowing us to apply an integrative systems biology work frame for biomarker development.

Our research focuses on defining the molecular events/network systems involved in the host‐pathogen interactions to identify biomarkers and therapeutic targets for pathogenic agents.

We apply systems approaches to provide a more complete understanding of the host‐pathogen interactions and pathways and networks regulated in both the host and pathogen during the course of infection and illness from various infectious diseases. Various types of ‐omics data are obtained to determine the kinetics of infection for exposures of human immune cells. These data are integrated with immunological, clinical and physiopathological data obtained from the animal models. Models are developed using the high‐throughput data to gain an overview of the host‐pathogen factors leading to the progression of the disease.
Replacement of aromatic amino acids based on their consensus temporal order suggests that ancient proteins tend to be intrinsically disordered
Authors: Meng Jingwei , Johnson Derrick , Oldfield Chris , Uversky Vladimir , Dunker A. Keith
Published: June 2021

The consensus temporal order of amino acids revealed that aromatic amino acids (F, Y and W) appear late in evolution. Interestingly, modern proteins that contains less aromatic amino acids show higher potential of intrinsically disordered properties. In this paper, we examined the hypothesis that ancient proteins with no aromatic amino acids are intrinsically disordered. Four Dephospho‐CoA Kinases (DPCK wild type, containing 5 to 19 aromatic amino acids) from 4 lower organisms were selected. All aromatics amino acids of these DPCKs were replaced with Leucine (DPCK L mutant, mimicking their ancestors) by whole gene synthesis and proteins were further purified by cobalt‐resin. The wild types were all soluble, while two of L mutants were only soluble with 4 M urea, suggesting these mutants are structurally unstable. The other two L mutants that are soluble also exhibited higher sensitivity to trypsin digestion and altered UV spectrum features of α‐helix and β‐sheet. Our results indicated that ancient proteins with less aromatic amino acids tend to be more intrinsically disordered. There are several advantages of this experimental observation. For example, intrinsically disordered proteins have less structure constraints and are more flexible to their binding partners. The impacts of these replacements to their kinase activity and specificity are under investigation.
HOW TO USE THE PSI STRUCTURAL GENOMICS KNOWLEDGEBASE TO ENABLE RESEARCH
Authors: Berman Helen M , Westbrook John , Gabanyi Margaret , Tao Yi‐Ping , Shah Raship , Schwede Torsten , Arnold Konstantin , Kiefer Florian , Bordoli Lorenza , Adams Paul , Carter Lester , Minor Wladek , LaBaer Joshua
Published: June 2021

The Protein Structure Initiative Structural Genomics Knowledgebase (PSI SGKB, URL: http://kb.psi‐structuralgenomics.org) is a web resource designed to turn the products of the structural genomics and structural biology efforts into knowledge that can be used by the biological community to understand living systems and disease. This workshop will present examples of how to use the PSI SGKB to enable biological research. For example, a protein sequence or PDB ID search will provide a list of protein structures from the Protein Data Bank, associated biological descriptions (annotations), homology models, structural genomics protein target information, experimental protocols, and the ability to order available DNA clones. A text search will find technology reports and publications that were created by the PSI's high‐throughput research efforts. Web tools that aid in bench top research, such as protein construct design, are also available. Created in collaboration with the Nature Publishing Group, the Structural Genomics Knowledgebase Gateway provides a research library, editorials about new research advances, news, and an events calendar also present a broader view of structural genomics and structural biology. The PSI SGKB is funded by the NIGMS.
Molecular recognition of FTase and GGTase‐I in vivo
Authors: Lamphear Corissa Lynn , Fierke Carol A.
Published: June 2021

Prenylation is an important post translational modification in which an isoprenyl tail is attached to the C‐terminus of a substrate protein, with protein farnesyltransferase (FTase) and protein geranylgeranyltransferase‐I (GGTase‐I) catalyzing the addition of a 15‐carbon farnesyl and 20‐carbon geranylgeranyl moiety, respectively. FTase and GGTase‐I have been proposed to recognize a canonical “CaaX” motif at the C‐terminus of protein substrates where “C” is cysteine, “a” is any aliphatic amino acid, and “X” is typically methionine, alanine, glutamate or serine for FTase and leucine or phenylalanine for GGTase‐I. However, recent studies indicate that the substrate selectivity of these enzymes may be more permissive with higher cross‐reactivity than originally proposed, raising the possibility that the cellular pool of prenylated proteins includes a wide range of C‐terminal sequences. In the human genome, ~700 proteins contain a cysteine four amino acids from the C‐terminus, but only ~70 proteins have been demonstrated to be farnesylated in vivo. To examine prenylation motifs in vivo, we created a library of GFP‐CaaX fusion proteins and are measuring the prenylation status using mass spectrometry and protein localization using fluorescence microscopy. These studies will provide insight into the in vivo specificity of prenyltransferases and the involvement of prenylation in various cellular processes.
Generation of O‐GlcNAc specific monoclonal antibodies using a novel synthetic immunogen
Authors: Teo Chin‐Fen , Ingale Sampat , Wolfert Margreet A. , Elsayed Galal A. , Not Laszlo G. , Chatham John C. , Wells Lance , Boons Geert‐Jan
Published: June 2021

O‐GlcNAc modification is a ubiquitous and dynamic post‐translational modification that has been implicated in many cellular processes, frequently via interplay with phosphorylation that can occur on the same serine and threonine residues. Unlike phosphorylation for which a wide range of pan‐ and site‐specific phospho‐antibodies are available, only two pan‐O‐GlcNAc specific antibodies have been described: an IgM pan‐O‐GlcNAc antibody and an IgG antibody raised against O‐GlcNAc modified components of the nuclear pore complex. In fact, it is well known that O‐GlcNAc modified glycoconjugates do not elicit relevant IgG isotype antibodies. Here, we report a novel method combining a fully synthetic three‐component immunogen with classical hybridoma technology to obtain a series of O‐GlcNAc‐specific IgG monoclonal antibodies. Large‐scale shotgun proteomics approach utilizing these antibodies led to the identification of more than 200 O‐GlcNAc modified proteins, of which approximately two‐thirds were novel. Application of this tool allowed us to delineate differentially modified proteins of the liver in response to hemorrhagic trauma and resuscitation in a rat model. This work was supported by grants from the NIDDK/NIH RO1 DK075069 (LW), NIH P41‐RR‐5351 (GJB), NIH RO1 CA88986 (GJB), NIH HL067464 (JCC) and NIH HL079364 (JCC). CFT is an AHA pre‐doctoral fellow (0715377B).
Quantitative phosphoproteomics of depolarization‐dependent protein phosphorylation in nerve terminals
Authors: Larsen Martin R. , Palmisano Giuseppe , Engholm‐Keller Kasper , Kulej Katarzyna , Xue Jung , Graham Mark , Robinson Phillip J.
Published: June 2021

Depolarization‐dependent calcium influx activates a number of kinases and phosphatases which result in altered phosphorylation of key proteins in, exocytosis, endocytosis etc. Only a small number of phosphosites in these proteins have been identified to date.

Here we have used iTRAQ labeling directly on TiO2 resin combined with the SIMAC strategy (Thingholm TE et al., MCP 2007), Hydrophilic interaction chromatography and high accuracy mass spectrometry to quantitatively assess phosphorylation sites in synaptosomes treated with KCL and low and high levels of 4‐aminopyridine (4AP).

Using the new quantitative phosphoproteomics strategy we have successfully identified more than 1500 phosphorylated proteins from highly pure synaptosomes. In these 1500 phosphoproteins we have identified and quantified more than 2500 phosphosites. More than 30 % of all the phosphosites change upon depolarization using either KCL or 4AP. In addition, we have identified 72 kinases and 19 phosphatases in synaptosomes whereof many of those are specifically regulated upon stimulation. Overall we show changes in the phosphorylation status of proteins participating in e.g., transportation of SVs, exocytosis and endocytosis.
Quantitative Chemical Proteomics of Dynamic Palmitoylation
Authors: Martin Brent , Cravatt Benjamin
Published: June 2021

Protein palmitoylation is an essential post‐translational modification necessary for trafficking and localization of numerous regulatory proteins that play key roles in cell growth and signaling. The commercially available fatty acid analog 17‐octadecynoic acid is readily metabolically incorporated in living cells at endogenous sites of protein palmitoylation. Labeled proteins can be chemically coupled to azide‐linked reporters for robust non‐radioactive analysis by multiple platforms, including fluorescent gel‐based detection and mass spectrometry‐based identification. This approach shows unprecedented sensitivity for profiling palmitoylated proteins in complex biological systems, leading to the identification of hundreds of palmitoylated proteins in human cell lines and mouse primary cells. In order to understand the processes regulating dynamic palmitoylation, we have developed a quantitative platform for global comparative proteomic analysis of palmitoylated proteins, including identification of exact sites of palmitoylation. This platform has been applied to interrogate the population of palmitoylated proteins regulated by both palmitoyl transferases and thioesterases by using novel small molecule inhibitors, shRNA knockdowns, and physiological stimulation. Funded by F32NS060559, CA087660 and the Skaggs Institute for Chemical Biology.
Identifying protease cleavage sites by label‐free mass spectrometry
Authors: Sun Qing , Gupta Nitin , Cieplak Piotr , Zhu Wenhong , Ratnikov Boris , Pevzner Pavel A , Smith Jeffrey W
Published: June 2021

Methods

a method for identifying the cleavage sites for proteases in complex proteomes is described. The method makes use of a computer program called MS‐Cleave. The utility of the approach is demonstrated by determining of the cleavage sites in a set of recombinant Escherichia coli proteins generated by two matrix metalloproteinases (MMPs), MMP‐9 and MMP25. MS‐Cleave analyzes LC‐MS/MS data of standard samples with no additional chemical labels, processes them through a stringent peptide identification procedure, filters out tryptic cleavages and terminal trimming products. Each cleavage site is given a score based on the cleavage efficiency.

Results

Twenty‐two E. coli proteins were subjected to proteolysis by either MMP‐9 or MMP‐25. The cleaved samples were combined and analyzed by LC‐MS/MS. The cleavage sites are validated by comparison with the known substrate specificity of the two proteases which were defined with a substrate phage display library.

Conclusion

MS‐Cleave analysis is a reliable and convenient approach for the identification of protease cleavage sites without the use of additional chemical tags. The method is applicable for any standard proteomics facility.

This work was supported by NIH 5U54RR020843‐05 grant to JWS, NIH 5R01RR016522‐05 and 1‐P41‐RR024851‐01 grants and Howard Hughes Medical Institute Professor Award to PP.
Selection of Aptamers against live E.coli Cells using Cell SELEX
Authors: Shipley Sheena LaShae , White Ethan , Kim Sung‐Kun
Published: June 2021

Aptamers are oligonucleotides that bind with great specificity and affinity to a target molecule. These oligonucleotides are produced through the course of Systemic Evolution of Ligands by Exponential Enrichment (SELEX). SELEX is a combinatorial chemistry technique used to generate a random DNA or RNA library, which is then incubated with a target molecule. The binding aptamers are divided from the nonbinding random pool DNA/aptamers, and then amplified via polymerase chain reaction (PCR). Double stranded DNA aptamer molecules have been used to select against purified target molecules; in this study we have developed a selection technique using live E. coli cells as a target and using Kluyveromyces lactis, Bacillus subtilis, and Enterobacter aerogenes as negative controls. Aptamer pools obtained from approximately 7–9 rounds of Cell SELEX demonstrated an affinity for E.coli cells when tested via flow cytometry and fluorescence detection.
Screen for Chemical Modulators of the Laminin‐induced Clustering of Dystroglycan Reveals Novel Inhibitors of Aquaporin‐4 Polarized Distribution in Astrocytes
Authors: Noel Geoffroy Pierre , Stevenson Sarah , Moukhles Hakima
Published: June 2021

Aquaporin‐4 (AQP4) is the principal water channel in the brain and is mainly concentrated at perivascular astrocyte endfeet. This distribution of AQP4 is of major importance because it renders the channel capable of regulating water fluxes between the brain and the bloodstream and therefore plays a role in the water homeostasis. Evidence points to a role of the dystrophin‐associated protein (DAP) complex in the localization of AQP4 at astrocyte endfeet, via dystroglycan (DG) interaction with perivascular laminin. Interestingly, lack of AQP4 concentration at these endfeet in the dystrophin null mouse has been associated with a delayed onset of brain edema. It is therefore of particular importance to identify molecules that modulate the perivascular localization of AQP4. Here, we carried out a high‐throughput screen, using a chemical library of 3,594 small molecules, to identify modulators of the laminin‐induced co‐clustering of the DAP complex and AQP4 in primary astrocyte cultures. Of the 6 inhibitory molecules identified, one induced the reduction of the clustering by promoting the metalloproteinase‐mediated shedding of DG. These findings demonstrate the robustness of our in vitro model that provides a powerful tool for high‐throughput screening and allows us to identify and validate new compounds capable of disrupting the laminin‐induced co‐clustering of the DAP complex with AQP4.

Grant Funding Source: Canadian Institutes of Health Research (CIHR), Muscular Dystrophy Canada (MDC)
Role of Heme in the Regulation of Signaling Kinases
Authors: Balamurugan Parimaladevi , Arvey Aaron , Yao Xiao , Huyn Donovan , Leslie Christina , Zhang Li
Published: June 2021

Heme, iron protoporphyrin IX, is arguably one of life's most central molecules. Many living organisms ranging from bacteria to humans can synthesize heme de novo. Heme not only serves as a prosthetic group in many enzymes and globins, but it also directly controls many regulatory and signaling processes in living organisms. In mammals, heme has shown to bind to and to regulate the activity of diverse signal transducers and regulators, including MAP kinases, the heme‐regulated inhibitor kinase HRI, the transcriptional regulator Bach1, and the circadian clock coordinator Rev‐erbα. Most, if not all, of these heme‐regulated proteins contain one or more heme‐responsive motifs containing two invariable CP residues. To explore the potential roles of heme in regulating diverse cellular processes, we employed computational methods to identify human proteins that contain one or more CP motifs. We found that many proteins with the heme‐binding CP motifs belong to regulatory and signaling networks such as tyrosine kinase networks. To ascertain the biological significance of these networks formed by potential heme binding proteins, we created HeLa cells with heme biosynthetic enzymes knocked down. We then examine the effect of heme on the activation of certain tyrosine kinases. Our data suggest that heme has the potential to control the activity of divers signaling pathways.
BIOLUMINESCENT AMPLIFICATION ASSAY FOR microRNA DETECTION
Authors: Sun Ye , Golovlev Val
Published: June 2021

We have previously reported a new sensitive and cost effective Reverse Transcription Quantitative Bioluminescent assay (RT‐qBLA) for detection and quantification of microRNA in biological samples1,2. The RT‐qBLA detects release of inorganic pyrophosphate when target microRNA is extended on specially designed DNA template. The proposed approach is fast and less expensive than microarrays and real‐time PCR for microRNA detection. It allows microRNA quantification in a broad dynamic range; requires less reagents per reaction; can be performed at a constant temperature and is amendable to various replication strategies including “constant temperature” amplification techniques such as Rolling Circle Amplification (RCA) and Strand Displacement Amplification (SDA). In this report we present development of a real‐time RCA and SDA for microRNA detection with enhanced detection sensitivity and specificity.
Acculturation factors and the proportion of older Mexican Americans with inadequate folate intakes: National Health and Nutrition Examination Survey (NHANES), 2003–2006
Authors: Hamner Heather Carter , Cogswell Mary E , Johnson Mary Ann
Published: June 2021

We examined the association of acculturation factors in Mexican Americans (MA) aged 60+ years with usual mean and inadequate folate intakes. We analyzed data in 1,695 non‐Hispanic white (NHW) and 466 MA adults aged 60+ years participating in NHANES 2003–2006. We estimated usual folate intakes accounting for within‐individual variation using data from two 24 hour diet recalls and average daily intake from supplements. Among MA, 45% spoke mostly Spanish and 42% were born in Mexico. Compared with NHW, MA who spoke mostly Spanish had lower mean usual folic acid intakes from fortified foods alone (103 μg vs.178 μg) and total folic acid (fortified foods + supplements) (189 μg vs. 413 μg), but not natural food folate (202 μg vs. 196 μg). Similar patterns were observed in MA who were born in Mexico. The proportion of NHW and MA adults aged 60+ years who consumed less than the Estimated Average Requirement (EAR) for folate was 10% and 17%, respectively; in MA who spoke mostly Spanish, the proportion was 26% and in MA who were born in Mexico, 22%. Post‐fortification, inadequate folate intake may be a concern among US MA aged 60+ years who speak mostly Spanish or were born in Mexico. Research funded by the Centers for Disease Control and Prevention.
Associations Between Folate, DNA Methylation, Methylenetetrahydrofolate Reductase Polymorphisms, and Colorectal Cancer Risk: A Review
Authors: Miszewski Susan Grace , Nazmi Aydin , Derelian Doris
Published: June 2021

A review of twelve studies showed consistent links between folate and colorectal cancer (CRC). Low dietary folate may lead to reduced levels of 5,10‐methylenetetrahydrofolate and accumulation of deoxsyuridylate in DNA which decreases DNA integrity and chromosomal breaks and increases CRC risk. Low dietary folate also leads to decreased DNA methylation; hypomethylation is suggested to inactive the p53 tumor suppressor gene. Folate deficiency produces strand breaks in human DNA, specifically in exons 5–8 of the p53 gene. These possible epigenetic factors depend on methylenetetrahydrofolate reductase genotype. Individuals with the 1298CC genotype showed lower risk of rectal cancer compared to those with the 1298AA or 1298AC genotypes. Individuals with the 677TT genotype showed reduced risk of rectal cancer when compared to those with the 677CT and 677CC genotypes. Levels of DNA methylation were consistently determined by folate status for those with the 677TT genotype, suggesting a possible role for plasma folate as an effect modifier among these individuals. Dietary folate status impacts risk for CRC via DNA methylation and chromosomal integrity, only in individuals with the 677TT genotype.

Source of Support: None
Paternal folate deficiency during reproduction influences folate concentrations in the brain of the offspring
Authors: Kim Hye Won , Lee Yo A , Kim Hwan‐Hee , Lee Suman , Chang Namsoo
Published: June 2021

This study was performed to investigate the effects of paternal folate status during reproduction on folate concentration and its possible influences on DNA methylation in the brain of the offspring. Experimental diet of a different folic acid content (0 vs. 8 mg/kg diet) was fed to animals of four groups; both parents supplemented (FSFS), paternal folate deficient (FDFS), maternal folate deficient (FSFD), and both parents deficient (FDFD). Experimental diets were fed for 4 weeks prior to mating and throughout the pregnancy. Brain folate concentrations were analyzed microbiologically and DNA methyltransferases by RT‐PCR, respectively. At day 20 of gestation, brain folate concentrations were significantly higher in FSFS (0.89±0.18) than in other 3 groups (FDFS; 0.49±0.06, FSFD; 0.42±0.08 and FDFD; 0.41±0.10 nmol/g wet tissue). Paternal folate deficiency resulted in a decrease of folate concentrations in a degree similar to maternal or both parental deficiencies. No significant difference was observed in the Dnmt 1, 3a and 3b expression level among the diet groups. Further studies are warranted to investigate the consequences of paternal folate deficiency on DNA methylation and gene expressions in relation to brain growth and development. This research was supported by Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science and Technology (2009‐0072246) and by BK21.
Selenium (Se) supplementation for 6 months increases plasma selenium levels in HIV+ adults in Botswana
Authors: Sales Sabrina , Tsalaile Lesedi , Burns Patricia J , Campa Adriana , Makhema Joseph , Marlink Richard , Baum Marianna K
Published: June 2021

Objective

To determine whether supplementing 200 mg/day of selenium (Se) for 6 months increases plasma Se in asymptomatic, ART naïve HIV+ adults in Botswana.

Methods

After consenting, 79 participants were randomized into the Se supplementation (N=33) or placebo (N=46). Fasting blood was drawn for plasma Se levels, CD4 cell count and HIV viral load. Se deficiency was defined as <0.085 mg/L.

Results

Mean age was 34.13±8.0 years and 72.2% were male. The prevalence of Se deficiency in this cohort was 94.9% (n=75). At baseline, there were no significant differences in mean Se levels between the two groups (0.065±0.010 mg/L vs. 0.070±0.024 mg/L, p=0.182). After 6 months of supplementation, there were significant differences in mean Se levels between the Se‐supplemented (0.147±0.153 mg/L) and not supplemented groups (0.069±0.121 mg/L, p<0.001). There were no significant differences in CD4 cell count and viral load between the groups at 6 months.

Conclusions

Supplementation with Se for 6 months normalized plasma Se in those who were deficient, and increased Se levels in all supplemented participants. This study also demonstrates that Se may be used as an index of adherence in micronutrient supplementation studies that contain Se. Further research is needed to determine the length of supplementation needed at this dose to reach steady state and observe clinical benefits of supplementation.

Funded by NIDA
Selenocysteine lyase expression in selenoprotein P knockout mouse
Authors: Seale Lucia A , Gilman Christy L , Hashimoto Ann C , Seyedali Ali , Berry Marla J
Published: June 2021

Selenium is an antioxidant micronutrient that is present in proteins as the amino acid selenocysteine (Sec). Sec is cotranslationally inserted into nascent polypeptides to form selenoproteins and selenium can be recycled from selenoproteins via degradation of Sec residues. Conformational similarities with prokaryotic β‐lyase suggest that selenocysteine lyase (Sec lyase) could be the eukaryotic enzyme involved in recycling the L‐selenocysteine residue from degraded selenoproteins, delivering the selenide byproduct back to the incorporation pathway. Two selenophosphate synthetases, SPS1 and SPS2, have been implicated in selenium utilization. Previous studies suggested that SPS2 synthesizes Sec from selenide and ATP and SPS1 is involved in selenium recovery via Sec lyase. A recent report described interaction of Sec lyase with both SPS enzymes. Our objectives are to characterize in vivo Sec lyase expression using a selenium‐depleted model, the SelP knockout mouse, and to investigate the relative functions of the two SPS enzymes under conditions of selenium limitation or sufficiency. Our results show differential regulation of Sec lyase mRNA levels in the mouse tissues. Effects of selenium status on interactions with SPS1 and SPS2 are under investigation. The evidence signals to a hierarchy of regulation of selenium utilization when selenium is limiting.
Effects of dietary Se deficiency or excess on gene expression of 13 novel selenoproteins in growing pigs
Authors: Zhao Hua , Zhang Qiaoshan , Li Ke , Tang Jiayong , Xia Xin‐Jie , Wang Kang‐Ning , Lei Xin Gen
Published: June 2021

We previously reported effects of dietary Se concentrations on gene expression of 12 selenoproteins in porcine tissues. This study was to determine responses of tissue mRNA levels for the rest of 13 out of 25 porcine selenoprotein genes to dietary Se deficiency and excess. Weanling male pigs (n = 30) were fed a corn (from Se‐deficient area)‐soy basal diet (0.02 mg Se/kg) supplemented with 0, 0.3, or 3.0 mg Se/kg as Se‐enriched yeast (Angel Yeast, Hubei, China) for 16 wk. In the end, samples of 11 different tissues were collected for relative mRNA level analyses using Real‐Time RT‐PCR. Compared with the Se‐supplemented pigs, the Se‐deficient pigs showed lower (P < 0.05) mRNA level of gpx1, gpx3, selpp1, selS in blood and several tissues. In contrast, dietary Se deficiency enhanced (P < 0.05) mRNA levels of sps2, selI, selO, selT and selV in various tissues. Pigs fed 0.3 mg Se/kg had greater (P < 0.05) mRNA levels of sel15, sps2, selH, selI, selM and selV in thyroid, spleen, or pituitary than those of the other two groups. The relative mRNA levels of selS and gpx1 in thyroid and selH and selI in pituitary were lower (P < 0.05) in pigs fed the Se‐deficient diet than those fed 0.3 mg Se/kg. In conclusion, the effects of dietary Se deficiency or excess on the gene expression of the 13 novel porcine selenoproteins in various tissues were non‐unilateral. (NSFC Projects 30628019, 30700585, and 30871844, and the Chang Jiang Scholars Program).
Weight loss is associated with increased serum 25(OH)vitamin D in overweight or obese women
Authors: Rock Cheryl L. , Heath Dennis D. , Flatt Shirley W. , Karanja Njeri , Pakiz Bilge , Sherwood Nancy E. , Thomson Cynthia A.
Published: June 2021

Circulating 25(OH)D concentration has been linked with differential risk and progression of disease. Obese individuals have lower circulating 25(OH)D, possibly due to sequestration in body fat. The objective of this study was to examine the effect of weight loss on 25(OH)D concentration in overweight or obese women in a clinical trial of a diet and lifestyle modification program. Study participants (n=442) were 44(10) (mean[SD]) yrs, with BMI 33.8(3.4) kg/m2, weight 92.1(10.7) kg, and waist circumference 108.6(9.6) cm. One year data are available for 92% of participants (n=406), and weight loss averaged − 9.7(7.7) kg for women assigned to intervention and − 2.8(6.8) kg for control groups, a relative weight loss of − 11% and − 3%, respectively. Total serum 25(OH)D was measured via chemiluminescent DiaSorin assay; 25(OH)D2 and 25(OH)D3 were separated and quantified with reversed phase HPLC methodology. Baseline 25(OH)D was 21(10) ng/mL, significantly associated with degree of obesity and race/ethnicity. In women who lost ≥10% of body weight at 12 mos, 25(OH)D increased to 23(10) ng/mL. In multivariate analysis, change in 25(OH)D over one year was significantly associated with degree of weight loss (P<.0001), adjusted for baseline level, age, and race/ethnicity. These findings suggest that weight loss promotes increased serum 25(OH)D in overweight or obese women. Funded by Jenny Craig, Inc.
Intakes and sources of vitamin D in the US population
Authors: Ahuja Jaspreet KC , Rhodes Donna , Goldman Joseph D , Omolewa‐Tomobi Grace , LaComb Randy , Moshfegh Alanna
Published: June 2021

Vitamin D critical due to its role in bone health has emerged as potentially playing an important role in other diseases. However, many knowledge gaps in the study of vitamin D exist, including knowledge of intake patterns from national surveys. The Food and Nutrient Database for Dietary Surveys 3.0 (FNDDS), based on USDA's National Nutrient Database for Standard Reference, both important components of nutrition monitoring in the U.S., were recently updated (2009) to include food composition values for vitamin D. FNDDS was applied to food intake data from the What We Eat in America, NHANES, 2005–2006 for 8,437 males and females, ages 1 and older, to provide nationally representative estimates of intakes of vitamin D from foods. The proportion of individuals with usual intakes above their Adequate Intake (AI) was determined using the NCI method. Food sources and major contributors of vitamin D were also determined using SAS version 9.2. The mean (SE) intake of vitamin D was 5.0 (0.12) μg/day. About one‐third of individuals 1 year and over met their AI. However, most individuals over 50 years, regardless of gender, did not meet the AI (15 μg/day). Major contributors include milk and milk products (52%), fish (9%), and ready‐to‐eat cereals (7%).
Anti‐oxidative effects of fermented rice bran extract on H2O2‐induced oxidative stress in 3T3‐L1 adipocytes
Authors: Kim Dongyeop , Jeong Bo Young , Fan Jiang Ping , Chung Hyun Chae , Kim Han sup , Ko You Hyuk , Kim Gyoung Woo
Published: June 2021

The purpose of this study is to investigate anti‐oxidative effects of fermented rice bran on oxidative stress model in 3T3‐L1 adipocytes. The Issatchenkia orientalis MFST1 is selected as the most optimal microorganism for fermentation of rice bran. To confirm whether FRB extract ameliorate insulin resistance related oxidative stress or not, that is evaluated by analyzing utilization of glucose, generation of reactive oxygen species (ROS), Oil red O stain, and expression of mRNA related to glucose homeostatsis and adipogenesis. FRB extract did not show a significant difference in glucose utilization assay, but FRB extract strongly inhibited generation of ROS and up‐regulated PPAR‐γ and adiponectin expressions. Ferulic acid (FA) as convincing potential anti‐oxidative compound of FRB extract was also tested in same way mentioned above. FA showed a significant inhibition of ROS generation and up‐regulation of adiponectin and GLUT4 related to glucose transportation and insulin sensitivity. This result suggests that FRB extract helps in enhancing the antioxidant capacity against oxidative stress by neutralizing the free radicals.
Effects of decaffeination process on the phenolic content and antioxidant capacity of brewed coffees
Authors: Freeman Michelle Ruth , Landin Sarah Elizabeth , Wilson Ted
Published: June 2021

Coffee is rich in phenolics which provide its color, taste and antioxidant capacity, although decaffeination may reduce coffee health benefits. Decaf methods, i.e. Swiss water (SW), supercritical CO2 (SCC), water:ethyl acetate(WEA), methylene chloride(MC), and water (H2O), may alter phenolic content, antioxidant capacity and health benefits. Decaf coffees were compared to regular caffeinated coffees (RC) of the same brand, bean, and roast. Phenolic content was measured in gallic acid equivalents (GAE in mg/L), RC: 143±2, SCC: 131±5, WEA:137±4, H2O: 146±5, MC 124±3, and SW 128±3(LSM ± SE). GAE of MC and SW were significantly lower than RC, whereas SCC, WEA, and H2O decaf methods did not affect GAE. Human LDL oxidation was induced with 10μM Cu++ in vitro and quantified (A234nm lag‐time) as % control in the presence/absence of diluted coffee. At 1:5,000 RC, SW, SCC, WEA, MC and H2O were protected LDL from oxidation vs control LDL oxidized without coffee. Antioxidant effects were less robust vs control at 10K and 50K dilutions. 5K dilutions of WEA, H2O, and MC had similar antioxidant capacities to RC, while SW and SCC were less than RC. TBARS confirmed RC and decaf coffee (5K) antioxidant capacities. This study suggests that some decaf methods may not alter phenolic content and antioxidant capacity. Decaf methods could be an independent factor for interpreting health benefit studies of decaf coffee.

Grant Funding Source: WSU Foundation
Anti‐inflammatory effects of docosahexaenoyl lysophosphatidylcholine in RAW 264.7 cells and in mice models
Authors: Kim MinJee , Sohn MiRan , Sok Dai‐Eun , Kim Mee Ree
Published: June 2021

Lysophosphatidylcholines(lysoPC) have been known to play a role as lipid mediators in various cellular responses. However, it is not clear whether polyunsaturated acyl group in the structure of lysoPC is required for the cellular function. Nonetheless, lysoPCs with polyunsaturated acyl group are known to be abundant in plant and animal sources. In this regard, we examined the anti‐inflammatory effects of lysoPCs. In this study, we examined the an ti‐inflammatory effects of various lysophosphatidylcholine (lysoPC) containing linoleoyl, arachidonoyl or docosahexaenoyl group in RAW 264.7 cells or in vivo. First, arachidonoyl lysoPC and docosahexaenoyl lysoPC were found to inhibit LPS‐induced nitro oxide production in RAW 264.7 cells, with IC50 values of 0.38 μM and 1.90 μM, respectively. However, pre‐incub ation with linoleoyl lysoPC up to 50 μM was not found to inh ibit the LPS‐induced nitro oxide production in RAW 264.7 ce lls. Idditionally, I.V. injection of docosahexaenoyl lysoPC suppres sed the Zymosan‐induced plasma leakage in ICR mice with IC50 value of 50 μg/kg and maximal inhibition around 80%. In contrast, arachidonoyl lysoPC was observed to inhibit Zymosan‐in duced plasma leakage in ICR mice less potently, and linoleoy l lysoPC (150 μg/kg) was failed to inhibit Zymosan‐induced pl asma leakage in ICR mice. In a separate experiment, where do cosahexaenoyl lysoPC peroxide and docosahexaenoic acid were tested for the effect on Zymosan‐induced plasma leakage, 17(S)‐hydroperoxy‐4,7,10,13,15,19‐docosahexaenoyl lysoPC and docos ahexaenoic acid were failed to inhibit t Zymosan‐induced plas ma leakage in ICR mice. From these results, it is proposed that d ocosahexaenoyl lysoPC displays potent anti‐inflammatory effects and this may participate in the regulation of LOX activity in biological systems.
Microarray analysis reveals effects on important biological pathways in the liver of mice fed a dietary soy isoflavone concentrate
Authors: Shay Neil F , Ricketts Marie‐Louise , Medrano Theresa I , Shiverick Kathleen T
Published: June 2021

Consumption of soy isoflavones is proposed to lower serum cholesterol levels, improve symptoms of type‐2 diabetes, and lower the incidence of certain types of cancer. This study provides further insight into the effects of a commercially‐available, FDA‐approved, dietary soy isoflavone concentrate (Novasoy® 400) on gene expression profiles in mouse liver. Nude mice, without tumors, were fed a control diet (Advanced Protocol verified casein diet 5K96, Test Diet, Richmond, IN) for 5 days and were then divided into two groups and fed either 1) control diet or 2) control diet supplemented with 0.5% Novasoy® for a further 7 days (n=5 per group). The livers were collected, RNA isolated and oligonucleotide microarray analysis performed using Agilent technology. The soy diet differentially up‐regulated 531 and down‐regulated 469 genes (P < 0.009). Up‐regulated genes were associated with the cell cycle, cell cycle control, cholesterol metabolism, lipid metabolism, lipid and fatty acid transport and detoxification. A significant down‐regulation in genes associated with the stress response, cell proliferation and differentiation were observed. These results demonstrate that a commercially available dietary soy isoflavone concentrate alters gene expression profiles associated with important metabolic pathways to contribute to the beneficial health effects associated with soy consumption.
Selective effect of dyslipidemic human sera on in vitro gene expression
Authors: Zirpoli Hylde , Caputo Mariella , Carraturo Antonio , Torino Gaetano , Fazio Alessia , Attya Mohamed , Rastrelli Luca , Tecce Mario Felice
Published: June 2021

Since serum lipids are affected by nutritional intake and endogenous metabolism, we assayed the ability of human sera, differing in several metabolic and nutritional parameters, to affect specific gene expression in human hepatoma cells using sera from hypercholesterolemic subjects (LDL‐cholesterol median is 260mg/dl) vs normolipidemic subjects (LDL‐cholesterol median is 160mg/dl).

Hypercholesterolemic sera decreased sterol regulatory element binding protein 1c (SREBP‐1c) mRNA by 40% (p<0.05). UDP‐glucuronosyltransferase 1A1 (UGT1A1) mRNA expression was significantly increased by hypercholesterolemic sera by 84% (p<0.01).

Determination of fatty acids levels in sera showed that arachidonic acid (AA) was 88% more abundant in high cholesterol subjects (p<0.01) while docosahexaneoic and eicosopentanoic acids (DHA and EPA), as quota of total detected fatty acids, were significantly higher in low cholesterol subjects by 25% (p<0.05) and by 80% (p<0.01) respectively. Higher DHA EPA and AA concentrations link to a 2 fold higher expression of UGT1A1.

Although individual serum components accounting for the observed effects cannot be distinguished, our data indicate that the serum cholesterol containing lipoproteins directly affect gene expression pathways involved in metabolic regulation.

This work was supported from the FARB 2007 and 2008 grants of the University of Salerno.

Grant Funding Source: Farb 2007/2008 grants of the university of salerno
Poly (ADP‐ribose) Polimerase (PARP) Inhibition in Experimental Acute Necrotizing Pancreatitis
Authors: Uysal Bulent , Yasar Mehmet , Oztas Emin , Sadir Serdar , Coskun Omer , Cayci Tuncer , Oter Sukru , Guven Ahmet , Korkmaz Ahmet
Published: June 2021

Introduction

Inhibition of poly(ADP‐ribose) polimerase (PARP) is shown to reduce inflammation in several entities. Thus, we designed a study to evaluate the efficacy of benzamide (B), a PARP inhibitor, in an experimental model of ANP.

Methods

Thirty Sprague‐Dawley rats were divided into 3 groups: sham, ANP, ANP+B groups. ANP was induced by infusing of 1 mL/kg of 3% sodium taurocholate into the common biliopancreatic duct. ANP+B group received 100 mg/kg/day benzamide i.p. for a total of 3 days after induction of ANP. The surviving animals were killed at the 4th day and their pancreas was harvested for biochemical, microbiological and histopathological analysis. Blood samples from animals were also obtained.

Results

In the ANP group, a clear increase in serum amylase and neopterin levels and tissue oxidative stress indices were present; almost all of these changes were found to be reversed near to their normal values in ANP+B group. Histopathologic injury scores were significantly high in ANP group (p<0.05, ANP vs. sham) which were significantly lower in ANP+B group (p<0.05, ANP+B vs. ANP). Evaluation of bacterial translocation presented significantly less infected sites in ANP+B group than in ANP animals (p<0.01).

Conclusion

Our data showed that inhibition of PARP with benzamide reduced the severity, the mortality, bacterial translocation rates and neopterin levels in the experimental ANP model of rats.