bluuebunny/crossref_metadata_2025_split huggingface dataset:
Effect of ZNT7 on the CD40 signaling pathway in B lymphocytes: a possible mechanism for a regulatory role of zinc in immune function
Authors: Huang Liping , Tepaamorndech Surapun , Oort Pieter , Kirschke Catherine P. , Cai Yimeng 
Published: June 2021

Zinc deficiency impairs the immune system leading to frequent infections. Although it is known that zinc plays critical roles in maintaining healthy immune function, the underlying molecular targets are largely unknown. In this study, we demonstrate that zinc is important for the CD154‐CD40‐mediated activation of downstream signaling pathways in human B lymphocytes. CD40 is a receptor localized on the cell surface of many immune cells, including B lymphocytes. When the T helper lymphocyte (Th) is activated by antigens, it expresses a membrane protein, CD154 that allows Th to bind to B cells via CD154‐CD40 interaction leading to B cell activation. We show that cellular zinc deficiency impaired the CD154‐CD40‐mediated p38 MAPK phosphorylation. Most importantly, we demonstrate that ZNT7 interacted with CD40. ZNT7 knockdown in B lymphocytes had a negative effect on the cell surface expression of CD40. Consequently, the CD40‐mediated p38 MAPK signaling transduction was down‐regulated in ZNT7KD B lymphocytes. Conversely, this p38 MAPK signaling activity was up‐regulated by over‐expression of ZNT7 in B lymphocytes. We conclude that prevention of zinc deficiency is a key to maintain an optimal function of immune system and CD40 is the molecule target for ZNT7 in regulation of immune function of B lymphocytes.Support or Funding InformationThis work was supported by the United States Department of Agriculture, ARS CRIS project 2032‐51000‐004‐00D.
Skin‐specific stearoyl‐CoA desaturase 1 deficiency protects against adiposity by enhancing IL‐6 expression
Authors: Dumas Sabrina N , Guo Chang‐an , Ntambi James M 
Published: June 2021

Stearoyl‐CoA desaturase 1 (SCD1), a lipogenic enzyme that adds a double bond at the delta 9 position of stearate (C18:0) and palmitate (C16:0) has been proven to be important in the development of obesity. Skin‐specific deficiency of SCD1 (SKO) provides protection against adiposity from a high‐fat diet, with improvements in glucose clearance, insulin sensitivity and hepatic steatosis. Curiously, these mice also display elevated levels of plasma interleukin‐6 (IL‐6) and increased white adipose tissue lipolysis. IL‐6 is a known mediator of white adipose tissue lipolysis. In the SKO mouse model, IL‐6 expression is evident in the skin sebaceous glands and around the hair follicles. Recently, IL‐6 has been connected to increased gluconeogenesis. While gluconeogenesis is paradoxical to the increased glucose clearance seen in SKO mice, a pyruvate tolerance test in the SKO mice confirmed a transient increase in glucose production. Therefore, we propose that skin‐derived IL‐6 leads to white adipose tissue lipolysis, which decreases the size of fat pads leading to decreased adiposity. In addition the release of triglycerides in adipose tissue provides the liver with acetyl‐CoA, NADH and glycerol necessary for hepatic glucose production.Support or Funding InformationSupported by USDA Hatch W2005
Dolicholphosphate Mannosyltransferase: A Glycosyltransferase with Unity in Molecular Diversities
Authors: Baksi Krishna , Zhang Zhenbo , Banerjee Dipak K 
Published: June 2021

N‐glycan provides structural and functional stability to glycoproteins and adds to their flexibility. Glycan biosynthesis is elaborative, multi‐compartmental and uses glycosyltransferases. Failure to assemble N‐glycan leads to phenotypic changes developing infection, cancer, congenital disorders of glycosylation (CDGs) among others. The biosynthetic process begins at the endoplasmic reticulum (ER) with the assembly of dolichol‐linked tetradecasaccharide Glc3Man9GlcNAc2‐PP‐Dol where DPMS plays a critical role. DPMS is also essential for GPI anchor biosynthesis, protein O‐mannosylation in yeast as well as O‐ and C‐mannosylation in eukaryotes. DPMS has been purified from several species and its gene has been cloned from 40 species (protozoan parasite to human). The sequence alignment indicates the presence of a cAMP‐dependent protein phosphorylation motif (PKA motif) in DPMS from 39 species. Hydrophobicity analysis of the DPMS sequence from bovine, human, S. crevisiae and A. thaliana all show the PKA motif each preceded and succeeded by a sequence of hydrophobic amino acid residues, i.e., the presence of a transmembrane domain. Irrespective of these similarities, the amino acid sequence at the C‐terminus and the kinetic behavior have classified DPMS as Type I and Type II enzyme. DPMS belonging to Type II sub‐class has been proposed to be a multimeric protein requiring DPM2 and DPM3 to anchor down DPM1 to the ER membrane. Based on the hydrophobicity as well as the successful expression of bovine DPM1 gene in E. coli we conclude that the logic proposing the above model needs to be revisited. Because of inherent diversities, DPMS has acquired sensing ability of cellular microenvironment and can serve as an excellent biosensor. In addition, DPMS activity is down regulated during aging but when upregulated in capillary endothelial cells due to activation of β‐adrenoreceptor angiogenesis, a hallmark of tumor growth is enhanced. Furthermore, DPMS from rat liver, rat parotid acinar cells, hen oviduct, bovine brain as well as from S. cerevisiae all showed increased activity upon phosphorylation in vitro.Support or Funding InformationThe work was partly supported by funds from NIH/NIMHD G12MD007583 (KB), NIH U54‐CA096297, Susan G. Komen for the Cure BCTR0600582, and NSF EPS‐1002410 (DKB).
Mannose moieties exhibit self‐adhesive interactions
Authors: Perera Komitige Hashanthi , Chandran Preethi L. 
Published: June 2021

Carbohydrate‐Carbohydrate interactions (CCIs) are self‐adhesive interactions, generally observed among proteoglycans and glycolipids expressed on the cell surfaces. Many biological processes such as embryogenesis, fertilization and metastasis are driven by CCIs. Mannoses are biologically‐significant carbohydrate moieties, that are extensively present on the surfaces of fungi and viruses, and known to mediate host‐pathogen interactions. We studied the adhesive interactions between mannobiose (disaccharides of mannose sugars)‐mannobiose molecules using atomic force spectroscopy (AFS).Bifunctional linkers were synthesized with thiol functional groups on one end and aryl‐azide functional group on the other. The linkers were self‐assembled on gold substrates (gold coated AFM sample disks and gold coated AFM probes) via the thiol end, and mannobiose was photochemically linked to the free aryl‐azide end of the surface‐anchored linkers. The mannobiose attachment on the substrates was verified via its selective binding to the lectin, concanavalin‐A.Mannobiose‐mannobiose molecules exhibited strong adhesive forces when probed with AFS in an aqueous medium. The force‐distance curves exhibited peak adhesive forces that subsequently ruptured in ‘step’ like patterns, indicating that the adhesive forces were due to well‐defined molecular interactions. The peak adhesive forces between mannobiose‐mannobiose molecules ranged from 0.3 to 5.7 nN. The final ‘step’ or rupture forces ranged from 0.1 to 1nN, with forces clustering at regular intervals which were multiples of 25 +/− 3pN. The latter suggests the likely magnitude of the single mannose‐mannose unbinding force to be 25 +/− 3pN. Lactose, a diastereoisomer of mannobiose, did not show self‐adhesive interactions. Free mannobiose reduced the mannobiose‐mannobiose adhesive forces in a concentration‐dependent manner; whereas free lactose was less effective in reducing the forces. Also, the mannobiosylated surface displayed ‘water structuring’ effects that extended tens of nanometers beyond the surface, and which were disrupted by increasing the salt concentration.This is the first study to report self‐adhesive function between mannobiosylated surfaces. The majority of fungi and virulent viruses such as HIV, Ebola, and Marburg have mannose patches on their surfaces, and this fundamental study might shed light on the biophysical role of these pathogen surfaces. The water‐structuring ability of clustered mannose molecules may manifest as a dense water shell around pathogens, to keep them hydrated and protected from host immune molecules.Support or Funding InformationThis work is supported by National Science Foundation under grant no. 1407891 awarded to Dr. Preethi Chandran and by a mini‐grant awarded to Dr. Preethi Chandran under NSF grant no. 1208880 (PI: Dr. Sonya Smith)
GRP78 is Neither Expressed on ER−/PR−/Her2− Human Breast Cancer Cell Surface nor Secreted
Authors: Serrano Jesus E , Romero‐Nutz Eva C , Sanchez Neysharie , Banerjee Aditi , Baksi Krishna , Banerjee Dipak K 
Published: June 2021

GRP78 (Mr78 kDa) is a glucose regulated protein. It is a member of the heat‐shock protein (HSP) family but located in the lumen of the endoplasmic reticulum (ER). Its function as ER chaperone translocating protein across the ER membrane that needs to be glycosylated at the asparagine residue present in the sequeon Asn‐X‐Ser/Thr (N‐linked glycosylation) is well recognized. This important biochemical event is essential for glycoprotein folding and function. For example, when the protein N‐glycosylation is impaired with tunicamycin a protein N‐glycosylation inhibitor, angiogenesis, a hallmark for tumor progression and metastasis is inhibited due to an induction of ER stress. Under such condition, GRP78 is overexpressed in microvasculature and in tumor tissue, concluding that GRP78 is a master regulator in ER stress induced unfolded protein response (upr)‐mediated apoptosis in tumor microvasculature (J. Biol. Chem. 286, 29127–29138, 2011; Pure Appl. Chem. 84, 1907–1918, 2012). This contradicts the current dogma that supports GRP78 expression on the tumor cell surface interfering with the therapeutic(s) and making them as tumor promoters instead. To evaluate the GRP78 localization, we have used ER−/PR−/Her2− (i.e., triple negative) human breast cancer cell line MDA‐MB‐231 (Caucasian) cultured normally or without serum as well as before or after inducing the ER stress. After establishing the expression of GRP78 mRNA by qPCR and protein by western blotting, we focused on its cell surface expression. Unfixed cells were stained with Concanavalin A (Con A; Specificity = α‐D‐Mannose, α‐D‐Glucose, branched mannose), wheat germ agglutinin (WGA; Specificity = (GlcNAc‐β‐(1,4)‐GlcNAc)1‐4>β‐GlcNAc‐NeuAc) as well as with anti‐GRP78 antibody followed by their detection by immunofluorescence microscopy using either Rhodamin or Alexa–conjugated secondary antibody. The fluorescence images were captured in a Zeiss microscope with Axiocam camera. Con A and WGA staining provided images of N‐glycans on the cell surface and supported the intactness of the cell membrane. On the other hand, GRP78 fluorescence was absent from the surface of these cancer cells. Similar results were also obtained irrespective of cells cultured in the absence of serum and/or in the presence of 1μg/mL of tunicamycin. GRP78 fluorescence however was detected in cells after either fixing them with ice‐cold methanol or after permeabilization with digitonin. In addition, western blotting also failed to detect GRP78 in the culture media of MDA‐MB‐231 cells. We, therefore, conclude that GRP78 is neither expressed on the outer‐leaflet of the cell surface of ER−/PR−/Her2− human breast cancer cells MDA‐MB‐231 nor secreted in the culture media.Support or Funding InformationSupported in part by NSF EPS‐1002410 (DKB) and NIH/NIMHD G12MD007583 (KB).
Acculturation and Hispanic‐Heritage Mothers' Perception of Child Weight as Related to Child Feeding Practices
Authors: Luna Viridiana , Villegas Elizabeth , Hannon Bridget , Hammons Amber , Cruz‐Santiago Michelle , Wiley Angela , Teran‐Garcia Margarita 
Published: June 2021

Hispanic children in the United States are disproportionately affected by obesity. Previous analysis revealed that mothers participating in the Abriendo Caminos intervention, aimed at preventing childhood obesity in Hispanic‐heritage families, incorrectly classified their overweight/obese child as normal weight. Effective obesity prevention efforts must consider mothers' roles in child feeding practices and their impact on child weight status over the life course. Acculturation and mother perception of weight may explain some child feeding practices. The aim of this study was to determine whether mothers' acculturation and perception of children's weight was associated with mothers' rationale for offering children snacks. Mother‐child dyads (n=53) of Mexican or Puerto Rican descent were recruited as part of the Abriendo Caminos randomized control/workshop multi‐state intervention. Here we report on the cross‐sectional baseline data acquired to date from participating mothers in the Illinois and California cohorts. The Brief Acculturation Rating Scale for Mexican Americans‐II (ARSMA‐II) was used for assessment of acculturation and a set of questions previously developed by Blaine and colleagues (Nutrients, 2015) was used to assess the rationale behind offering snacks. Anthropometric measures were collected from all participants. One‐way ANOVA and Fisher's LSD were used to compare differences in mean times per week that snacks were offered. At baseline, 64% of children were overweight or obese, while only 24% of mothers categorized their child as such. Over half of the mothers (66%) were categorized with low levels of acculturation (very Hispanic oriented). There were no significant associations between child BMI, mother perception of child weight and reasons mothers offered snacks. Compared to mothers who were the least acculturated, those who were the most acculturated reported offering their child more snacks per week because their child was hungry (4.00, p < 0.05). A further understanding of how weight perceptions and acculturation influence health and feeding practices can equip professionals to provide culturally relevant tools for mothers to enact positive changes in their families.Support or Funding InformationThis research was supported by the Agriculture and Food Research Initiative Competitive Grant no. 2015‐68001‐23248 from the USDA National Institute of Food and Agriculture.
Dietary Long‐chain PUFAs Enrich Porcine Alveolar Macrophages and Alter Prostaglandin E2 Production Following LPS Challenge
Authors: Walter Kathleen R , Xi Lin , Jacobi Sheila K , Esposito Debora , Odle Jack 
Published: June 2021

The US swine industry experiences severe production and profit losses from respiratory infections despite the wide spread use of vaccines and antibiotics. Long‐chain PUFAs can modulate immune cell function through the production of eicosanoids, such as prostaglandin E2 (PGE2) which is capable of stimulating both inflammation and inflammatory resolution upon infection. The aim of this study was to determine the effects of dietary arachidonic acid (ARA) and eicosapentaenoic acid (EPA) on cultured porcine alveolar macrophages stimulated with LPS. Day‐old pigs (n=60) were fed milk replacer for 21d. Diets contained 0.5% ARA (control), 2.5% ARA or 3.5% EPA of total fatty acids. Alveolar macrophages were isolated via bronchoalveolar lavage, cultured and stimulated with LPS for 24h. Total fatty acids in alveolar macrophages and lung tissue were analyzed via gas chromatography‐mass spectrometry (GC‐MS). Alveolar macrophages were cultured and stimulated with 10ng/mL LPS or unstimulated (control). Concentrations of PGE2 in tissue culture media were measured via competitive enzyme‐linked immunosorbent assay (ELISA) kits. Average body weight and clinical hematology were unaffected by diet treatments (P >0.5). Concentration of ARA (% of fatty acids, w/w) in alveolar macrophages from pigs fed 2.5% ARA had an 82% increase in ARA compared to controls, and the concentration was decreased 21% in pigs fed 3.5% EPA compared to control (P < 0.01). Lung parenchymal tissue showed a similar enrichment pattern; tissue was enriched by 39% in pigs fed 2.5% ARA, while pigs fed the 3.5% EPA diet had a 45% decrease in ARA compared to control (P < 0.002). Media concentration of PGE2 was increased following LPS stimulation of alveolar macrophages. The increase in PGE2 concentration was greater in macrophages enriched in ARA, exceeding that from control cells by 187% and that from EPA‐enriched cells by 191% (P < 0.05). These data demonstrate that dietary supplementation of LC‐PUFAs can effectively alter lung tissue and alveolar macrophage fatty acid composition. Furthermore, these data validate that increased dietary ARA is an effective means to increase eicosanoid production by LPS‐stimulated immune cells.Support or Funding InformationUSDA Animal Health Funds
In vitro Hypoallergenic Potential of Peanut Protein‐Polyphenol Aggregate Particles
Authors: Plundrich Nathalie J. , Bansode Rishipal R. , Williams Leonard L. , Lila Mary Ann 
Published: June 2021

The single most effective management of peanut (PN) allergy is the strict avoidance of the allergenic food. However, PN proteins are found in a variety of food products. A lot of effort has focused on mediating PN allergy either through attempted modulation of the human immune system (e.g. oral immunotherapy) or the allergenic food proteins (by processing techniques or genetic modification). In this study, we took advantage of the natural binding properties of plant‐derived compounds (i.e. polyphenols) to proteins. We created stable aggregate particles comprised of PN proteins and cranberry pomace or PN skin polyphenols which were investigated for their IgE binding potential. Their capacity to modulate mast cell degranulation and cell signaling involving phosphorylation of MAPK and p‐38 was investigated in vitro using RBL‐2H3 cells.PN protein‐cranberry pomace and PN protein‐PN skin polyphenol aggregate particles were created by complexing cranberry (Vaccinium macrocarpon Ait.) pomace or PN skin polyphenols with roasted PN flour. Particles containing 5 to 40% polyphenols were created. For the detection of PN‐specific IgE binding by PN proteins, immunoblotting was performed with pooled plasma from seven PN‐allergic individuals. RBL‐2H3 mast cells were exposed to aggregate particles or PN flour and evaluated for their inhibitory activity on ionomycin‐induced degranulation (β‐hexosaminidase and histamine).Results showed that PN protein‐bound polyphenols were able to decrease the IgE binding capacity to PN proteins in aggregate particles and the highest reduction was observed in the 40% polyphenol samples, a 22% and a 38% decrease in the PN protein‐PN skin and cranberry pomace polyphenol aggregate particles, respectively. This was significant when compared to IgE binding capacity of unmodified PN proteins (p<0.05; n=4 or 5). Anti‐DNP IgE‐sensitized RBL‐2H3 cells challenged with DNP‐BSA and ionomycin in the presence of aggregate particles were evaluated for their release of β‐hexosaminidase and histamine. PN protein‐PN skin and cranberry pomace polyphenol aggregate particles appeared to reduce ionomycin‐induced β‐hexosaminidase release (60 and 61%, respectively) and histamine release (64 and 66%, respectively). PN protein‐PN skin and cranberry pomace polyphenol aggregate particles when exposed to ionomycin‐induced RBL‐2H3 cells, inhibited phosphorylation of MAPK and induced the phosphorylation of p‐38 in a dose dependent manner.Results suggest the modification of PN proteins with cranberry pomace and PN skin polyphenols led to the formation of PN protein‐polyphenol aggregate particles with reduced allergenic potential. Future trials are warranted to investigate the immunomodulatory mechanisms of these aggregate particles.Support or Funding InformationThis study was funded through USDA NIFA
US and Canada Joint Effort for Dietary Reference Intake Updates
Authors: de Jesus Janet , Klurfeld David M , Lynch Christopher J. , Brooks Stephen P 
Published: June 2021

Representatives from the US and Canadian Government will present updates from the Joint US and Canada Dietary Reference Intake (DRI) Working Group on activities conducted to work toward updating the sodium and potassium DRIs. The Governments of the United States and Canada have jointly undertaken the development of the Dietary Reference Intakes since the mid‐1990s. Federal DRI committees from each country work collaboratively to identify DRI needs, prioritize nutrient reviews and advance work to resolve any methodological issues that could impede new reviews. The commission of a systematic review for nutrients under review is now an integral part of the DRI process. Recently the DRI Working Group sponsored an evidence review by the Agency for Health Care Research and Quality (AHRQ) on sodium and potassium to inform the update of the sodium and potassium DRIs by the National Academy of Medicine (NAM). The AHRQ review will be completed in 2017. Additionally, the DRI steering committees jointly decided that prior to undertaking a nutrient review, the scientific framework for setting DRIs needed to be adapted to include a framework for including data on chronic disease risk reduction. Thus, a scientific expert panel was convened to review and critically evaluate evidentiary, dose response, and process issues related to the use of chronic disease endpoints and develop options for their incorporation into future DRI reviews. The Joint US/Canadian‐sponsored Working Group's report ‘Options for Basing DRIs on Chronic Disease Endpoints’ will inform the NAM committee that will develop guiding principles for inclusion of chronic disease endpoints to be used by committees setting future DRIs. In summary, the US and Canadian Governments are working collaboratively to adapt the methods for updating the DRIs utilizing rigorous systematic evidence reviews and a chronic disease endpoint framework.Support or Funding InformationUnited States Department of Health and Human Services, United States Department of Agriculture, and Health Canada.