bluuebunny/crossref_metadata_2025 huggingface dataset:
Effect of inhibitor of sphingolipid synthesis on Leishmania (Viannia) braziliensis cytokinesis
Authors: Straus Anita H , Castro Erica V. , Takahashi Helio K.
Published: June 2021

Leishmania (Viannia) braziliensis is a parasite widely distributed in Brazil and the causative agent of cutaneous leishmaniasis and occasional mucosal or mucocutaneous disease. The role of L. braziliensis sphingolipids was analyzed using myriocin, an inhibitor of serine palmitoyltransferase. Myriocin 5 μM significantly reduced synthesis of inositol phosphorylceramide (IPC), as quantified by high performance thin layer chromatography and electrospray ionization mass spectrometry. Myriocin treated promastigotes displayed a variety of aberrant cell phenotypes. The percentage of cells with one nucleus and one kinetoplast (1N1K), decreased from 89% (control) to 27% or 3% following treatment with 1 μM or 5 μM myriocin, respectively. High percentage of myriocin treated parasites exhibited large atypical cells presenting three or more nuclei (32% and 89% for 1 μM or 5 μM myriocin, respectively). It is noteworthy that addition of 2 μM 3‐ketodihidrosphingosine, a long‐chain base precursor of ceramide, restored IPC synthesis and parasite morphology. When L. braziliensis treated with 1 μM myriocin were analyzed by transmission electron microscopy it was observed 4N parasites, possibly as a result from an incomplete cytokinesis. Our findings indicate that sphingolipids are essential for completion of cytokinesis and may play a major role in L. braziliensis cell proliferation. Supported by FAPESP, CNPq, CAPES.
Characterization of the in vitro activity of the Arabidopsis thaliana lysoglycerophospholipid acyltransferase, At1g78690
Authors: Freeman Daniel A , Garrett Teresa A
Published: June 2021

The Arabidopsis thaliana enzyme At1g78690 is an acyltransferase that acylates 1‐acyllysophosphatidylglycerol or 1‐acyllysophosphatidylethanolamine (lyso‐PE) to phosphatidylglycerol or phosphatidylethanolamine (PE), respectively, in vitro. However, the over expression of the protein in Escherichia coli leads to the accumulation of the headgroup‐acylated lipid acylphosphatidylglycerol (APG) in vivo. In order to understand how the acylation of lyso lipids impacts the metabolism of APG, we are investigating in depth the in vitro reaction of At1g78690. A N‐terminal, 6‐histidine tagged At1g78690 was overexpressed in E. coli and then cell lysates fractionated to yield cytosol and membranes. The At1g78690 activity, assessed by monitoring the transfer of 14C‐labeled palmitate from palmitoyl‐CoA to lyso‐PE, fractionates nearly equally between the cytosol and membrane fractions indicating that At1g78690 is a peripheral membrane protein. Varying the in vitro reaction conditions has shown that EDTA and EGTA stimulate activity. Furthermore, MgCl2 at concentrations <10 mM inhibit At1g78690 activity. Preliminary trials with inclusion of NaCl showed improved activity. Assessments of the pH optimum and detergent dependence, as well as purification of At1g78690 are underway. This work was supported by a National Science Foundation‐Research at Undergraduate Institution Award #1152463.
Casein kinase II phosphorylation of yeast Pah1p phosphatidate phosphatase
Authors: Hsieh Lu‐Sheng , Carman George M
Published: June 2021

The phosphatidate phosphatase (PAP) enzyme catalyzes the dephosphorylation of phosphatidate to yield diacylglycerol and Pi. The diacylglycerol product is used for the synthesis of triacylglycerol as well as the major phospholipids phosphatidylcholine and phosphatidylethanolamine. In yeast, Pah1p PAP is subject to phosphorylation and dephosphorylation, and this regulation governs the cellular location of the enzyme. The phosphorylated enzyme is primarily associated with the cytosol, whereas the dephosphorylated enzyme is primarily associated with the membrane where the substrate phosphatidate resides. We examined the phosphorylation of Pah1p PAP by casein kinase II, a serine/threonine protein kinase highly conserved in eukaryotic organisms and essential for cell viability in yeast. Pah1p was a bona fide substrate for casein kinase II (e.g., kinase activity was dependent on time and enzyme concentration and dependent on the concentrations of Pah1p and ATP). Casein kinase II phosphorylated Pah1p on multiple serine and threonine residues, which was accompanied by the inhibition of PAP activity. A combination of mass spectrometry and site‐directed mutagenesis followed by phosphopeptide mapping analysis was used to identify Ser‐313 and Ser‐705 as major phosphorylation sites. Additional sites are being identified by the same approaches. Supported by NIH grant GM‐50679.
Yeast Dgk1p diacylglycerol kinase is regulated by the transcription factor Reb1p
Authors: Qiu Yixuan , Fakas Stylianos , Carman George M
Published: June 2021

In the yeast Saccharomyces cerevisiae, Dgk1p diacylglycerol (DAG) kinase catalyzes the CTP‐dependent phosphorylation of DAG to form phosphatidate. The enzyme activity is required for phospholipid synthesis during growth resumption from stationary phase when de novo fatty acid synthesis is inhibited. Growth resumption is accompanied by the induction of DAG kinase activity, which implicates a potential transcriptional regulation of DGK1. The promoter of DGK1 contains a consensus sequence TTACCCG (−167 to −173) for the binding of the transcription factor Reb1p. Using purified recombinant His6‐Reb1p, we performed electrophoretic mobility shift assays to examine the interaction of Reb1p with the potential Reb1p binding site. The interaction of His6‐Reb1p with a radiolabeled oligonucleotide probe was dependent on the concentration of His6‐Reb1p, and non‐labeled probe competed with this interaction. Mutations in the Reb1p binding site reduced this interaction. The DGK1 gene with a mutation in the Reb1p binding site (DGK1reb1) was expressed in dgk1Δ mutant cells and caused a low constitutive level of DGK1 mRNA, Dgk1p, and DAG kinase activity. In addition, the dgk1Δ cells expressing DGK1reb1 showed a defect in growth resumption from stationary phase and the mobilization of triacylglycerol for membrane phospholipid synthesis was compromised. Supported by NIH grant GM‐28140.
The C‐mannosyltransferase
Authors: Bakker Hans , Buettner Falk F.R. , Ashikov Angel
Published: June 2021

C‐mannosylation of tryptophan residues occurs on proteins containing the consensus sequence WXXW, particularly thrombospondin repeat containing proteins and type I cytokine receptors. Mutagenesis of the WXXW motif in different proteins had already established important roles of this motif, suggesting that protein C‐mannosylation is as vital as N‐or O‐glycosylation. However, the genetic base of the enzyme catalyzing C‐mannosylation was so far not known.

We identified C. elegans DPY‐19 as a protein with low sequence identity to the enzyme responsible for N‐glycosylation (oligosaccharyltransferase, OST). Homology is mainly based on identical overall membrane topology. Few amino acids are conserved, but three of these have been suggested to be involved in lipid linked donor‐substrate recognition of OST.

We demonstrated that DPY‐19 is a C‐mannosyltransferase. Interestingly, the gene encoding DPY‐19 has long been known from a mutant presenting a defect in neuroblast migration. We showed that the dpy‐19 mutant is devoid of C‐mannosyltransferase activity. Moreover, by recombinant expression of DPY‐19 and mass spectrometric analysis, we could show C‐mannosylation of target proteins such as UNC‐5 and MIG‐21. Remarkably, dpy‐19 and mig‐21 mutants present identical phenotypes in neuroblast migration. Thus, our work showing C‐mannosylation of MIG‐21 by DPY‐19 provides a functional link between both mutants.
O‐GlcNAcylation of nuclear proteins in the protozoan parasite Toxoplasma gondii
Authors: de Monerri Natalie Clare Silmon , Che Fa‐Yun , Calloway Myrasol , Nieves Edward , Nardelli Sheila Christina , Angeletti Ruth Hogue , Weiss Louis M. , Wu Peng , Kim Kami
Published: June 2021

The protozoan pathogen Toxoplasma gondii is able to switch between two developmental stages: tachyzoites, which replicate rapidly and cause severe disease, and dormant, cyst‐forming bradyzoites, which persist in organs and show a tropism for central nervous tissue such as the brain. Switching is central to disease pathogenesis and is thought to result from the ability of parasites to sense environmental changes, including nutrient availability. The transcriptional and protein profiles of tachyzoites and bradyzoites differ significantly, but little is known about how these changes are regulated at the genetic and epigenetic levels. Generally, posttranslational modifications play an important role in modulating gene expression by modifying nuclear proteins. O‐GlcNAcylation, the addition of beta‐D‐N‐acetylglucosamine (O‐GlcNAc) to Serine or Threonine residues, is a dynamic modification that occurs on many nuclear and cytoplasmic proteins. It was recently shown to occur on histones and is involved in stress responses in several organisms. Here, we provide evidence by chemoenzymatic, immunological and mass spectrometric methods that this modification occurs on nuclear proteins in T. gondii and furthermore, that its occurrence changes upon induction of cellular stress. We propose that O‐GlcNAcylation plays a role in regulating gene expression during tachyzoite‐bradyzoite differentiation.
Regulation of Gene Expression by Short Poly‐N‐Acetyl Glucosamine (sNAG) Nanofibers in Endothelial Cells and Keratinocytes
Authors: Vournakis John , Amemiya Yutaka , Bacopulos Stephanie , Zhang Aiguo , Muise‐Helmericks Robin , Demcheva Marina , Seth Arun
Published: June 2021

O‐GlcNAcylation is a catalyzed transfer of N‐acetylglucosamine to protein substrates. This modification aids cells in regulation of many crucial cellular processes. Short poly poly‐N‐acetyl glucosamine nanofibers (sNAG) enhance wound healing, by cell migration, and hemostasis, by promoting fibrin matrix and clot formation.

This study investigated the effects of sNAG on global gene expression using the SOLiD 5500xl Next generation sequencing system in both HUVEC (Human Umbilical Vein Endothelial Cells) and HaCaT (Human keratinocytes) focussing on those having roles in angiogenesis and wound healing.

In HUVEC cells, 1,705 genes had at least 2‐fold up‐or down‐regulation upon sNAG treatment. Genes which remained up‐regulated at both 1 and 6 hours of treatment were NFKBIZ, defensins and wound heaIing responses. Many of the collagen family of proteins, E‐Selectin, ICAM1, and cytokines, CXCL1 and CCL2, were up‐regulated transiently.

In HaCaT cells, 2,329 genes showed 2‐fold up regulation in response to sNAG (1 or 6 hours) and 2,487 genes showed 2‐fold down regulation. Genes that were up‐regulated over the 6 hour treatment include desmogleins and keratins, suggesting regulation of keratinocyte differentiation and re‐epithelialization.

Responses to sNAG observed in this study add support to its role in angiogenesis and wound healing pathways at the level of regulation of gene expression.
Osteoglycin (OGN) modulates inflammation during viral myocarditis via an interaction with Toll Like Receptor 4.
Authors: Rienks Marieke , Papageorgiou Anna Pia , Verhesen Wouter , van Leeuwen Rick , Carai Paolo , Westermann Dirk , Heymans Stephane
Published: June 2021

Background

Small leucine rich proteoglycans, like OGN, present in the extracellular matrix, have been described to interact with collagen and soluble growth factors, thereby modifying the extracellular milieu. Not much is known however about their influence on inflammation, therefore we investigated the role of OGN in viral myocarditis.

Results

In both humans and mice with viral myocarditis, we demonstrated increased expression of OGN. We showed distinct OGN protein footprints in different murine cardiovascular disease models, with, a of 72kDa variant most abundant in viral myocarditis. After treatment with chondroitinase ABC there was a shift in detected size indicating the attachment of chondroitin sulfate to OGN. The 72kDa OGN co‐immunoprecipitated with TLR4. Lastly, stimulation of bone marrow derived macrophages with LPS demonstrated a decrease in cytokine production, in the presence of OGN.

Conclusion

We demonstrated for the first time that variants of OGN are present in different cardiomyopathies. We showed that the OGN variant abundant in viral myocarditis had the glycosaminoglycan chondroitin sulfate attached to it, and that this variant bound to TLR4, hence modulate the response of macrophages to LPS. Understanding this mechanism and identifying the biochemical substrates responsible for this interaction, could lead to much needed new therapies for viral myocarditis.
Characterization of the Substrate Specificity of Mammalian Heparanase Using a Glycomic Approach
Authors: Mao Yang , Huang Yu , Wu Zhengliang , Zaia Joseph
Published: June 2021

Heparanase is an endo‐β‐D‐glucuronidase that catalyzes the partial depolymerization of Heparan sulfate (HS) glycosaminoglycans and plays a central role in the remodeling of the HS containing extracellular matrix. The activity of heparanase directly correlates with the metastatic potential of tumor‐derived cells. Here we present a glycomic approach to study the substrate specificity of heparanase by LC‐MS profiling the structure features of the heparanase‐cleaved products, which carry information about the heparanase cleavage sites. Our results indicates that heparanase cleaves near the boundary of the highly sulfated domains of HS. The LC‐MS profiles are consistent across the different HS substrate from different sources, confirming that they represent the substrate specificities of heparanase. We further studied the detailed structures of the most abundant oligosaccharides by tandem MS. The structural features of those oligosaccharides are in good agreement with previous studies on the substrate specificities of heparanase using other approaches.
HYALURONAN SYNTHESIS IS REGULATED BY INTRACELLULAR O‐GLCNACYLATION OF HAS 2
Authors: Passi alberto , Deleonibus Sara , Vigetti Davide , Viola Manuela , Karousou Evgenia , De luca Giancarlo
Published: June 2021

The amount of hyaluronan (HA) often reflects the progression of the diseases involving extracellular matrix (ECM) as it promotes neo‐angiogenesis, cell migration and inflammation and its synthesis is regulated by several factors as the phosphorylation of synthetic enzyme HAS2. In this work, based on human smooth muscle cell model, we studied the intracellular regulation of HA synthesis at molecular level. The hexosamine biosynthetic pathway (HBP) may increase the concentration of HA precursor UDP‐Nacetylglucosamine leading to an increase of HA synthesis.

We found that the inhibition of O‐GlcNacylation strongly reduced HA production whereas treatments that induced protein O‐GlcNacylation increased HA secretion. Gene expression studies done by quantitative RT‐PCR revealed that HAS2 mRNA was the most sensible to O‐GlcNacylation and accumulated after its induction. We found that the transcription regulator YY1 activates HAS2 expression after O‐GlcNacylation. Using IP and recombinant 6myc‐HAS2 we demonstrate the HAS2 residue O‐GlcNacylated and we quantified AoSMCs motility and monocytes binding and found that O‐GlcNacylation increased cell invasion and inflammatory cell recruitment. Moreover, using ChIP/sec approach we demonstrated that the O‐GlcNacylation is an epigenetic modification of protein interacting with HAS2 promoter.
Thyroid hormone induction of adult stem cell formation during postembryonic development: An essential role of sonic hedgehog signaling mediated cell‐cell interaction
Authors: Shi Yun‐Bo
Published: June 2021

Adult organ‐specific stem cells are essential for organ homeostasis and tissue repair and regeneration, but the underlying mechanism for their development is unclear. Intestinal remodeling during frog metamorphosis offers a unique opportunity to study the formation of such stem cells during vertebrate development. During the transition from an herbivorous tadpole to a carnivorous frog, the intestine is completely remodeled with the larval epithelial cells undergo apoptosis and are replaced by adult epithelial cells developed de novo. The entire metamorphic process is under the control of thyroid hormone. We have shown that adult epithelial stem cells are induced by thyroid hormone through dedifferentiation of some larval epithelial cells. Interestingly, thyroid hormone action in larval epithelial cells alone can induce the formation of putative precursors of the adult stem cells, the formation of the stem cells and their eventual differentiation into the adult epithelium require thyroid hormone action in the nonepithelial tissues as well. This latter action is likely through the connective tissue, leading to the formation of proper stem cell niche. We will further provide evidence that sonic hedgehog signaling, which is induced by thyroid hormone, is one important pathway that mediates the cell‐cell interaction for the formation of the adult intestinal stem cells during this process.
ERK MAP Kinases Are Critical for the Expression of MKP‐1 and MKP‐2 in Macrophages Following LPS Stimulation
Authors: Liu Yusen , Xue Jianjing , Shakibi Yasaman , Xu Pingping , Wancket Lyn M. , Nelin Leif D. , Frazier W. Joshua
Published: June 2021

Although the MAP kinase phosphatase (MKP) family plays an important role during host defense, the regulation of MKPs during the immune response is not well understood. We studied the expression of MKP‐1 and MKP‐2 in RAW264.7 macrophages during LPS stimulation. We found that pretreatment of cells with U0126, a pharmacological inhibitor of MEK1/2, substantially decreased MKP‐1 and MKP‐2 mRNA levels in LPS‐stimulated macrophages. In contrast, a p38 MAP kinase inhibitor, SB203580, had little effect on the levels of MKP‐1 and MKP‐2 mRNA. The combination of U0126 and SB203580 resulted in greater decreases in MKP‐1 and MKP‐2 mRNA levels than did U0126 alone, suggesting that p38 contributes to MKP‐1 and MKP‐2 induction in response to LPS. Interestingly, in the absence of LPS, SB203580 enhanced ERK and MEK1/2 activities as well as MKP‐2 mRNA and protein expression, suggesting that p38 negatively regulates the ERK pathway by inhibiting a signaling event upstream of MEK1/2. LPS stimulated a potent CREB phosphorylation through a process dependent on p38, but not ERK, and enhanced the localization of phospho‐CREB to the chromatin of both MKP‐1 and MKP‐2. At the protein level, both MKP‐1 and MKP‐2 underwent robust phosphorylation mediated by ERKs. Taken together, our studies suggest that the ERK pathway plays a predominant role in the regulation of MKP‐1 and MKP‐2 through both transcriptional and post‐translational mechanisms.
Potential SET protein involvement in autophagy in cancer
Authors: Ouchida Amanda Tomie , Uyemura Valéria Tudella , Leopoldino Andréia Machado , Curti Carlos
Published: June 2021

Autophagy is a process by which lysosomes degrade and recycle long‐lived proteins and cellular organelles. It degrades cellular components that are worn out/damaged or needed to generate substrates to maintain cellular energy homeostasis under conditions of limited nutrients or stress. Cells of squamous cell cancer of the head and neck (HNSCC), which is the fifth most common malignancy in the world, have a high expression of the SET protein and under oxidative stress they perform the autophagic process. Therefore, the aim of our study is to verify a potential involvement of the SET protein on the autophagic process in these cells exposed to oxidative stress. Initially, knockdown of the SET protein (siSET) was performed in the Cal 27 cell line, in which oxidative stress was induced with tert‐butylhydroperoxide (t‐BHP). Proteins involved in autophagy as LC3B, Beclin‐1 and p62 were analyzed by immunoblotting. The siSET in Cal 27 cells caused an increase in the level of these proteins, which was slightly and significantly reduced after 4 and 8 hours of exposure to t‐BHP, respectively. Acids vacuoles identified by immunofluorescence, which are indicative of autophagy, were also found in Cal 27 cells for siSET and treatment with t‐BHP. Therefore, we believed that SET protein is important for the modulation of autophagy in HNSCC. PCR array assays are now in progress in order to assess potential genes involved in this regulation.

FAPESP and CAPES
VDAC phosphorylation impairment causes mitochondrial membrane permeability during lipid accumulation
Authors: Martel Cécile , Fanelli Elena , Boursier Céline , Henry Céline , Calamita Giuseppe , Lemoine Antoinette , Brenner Catherine
Published: June 2021

Non‐alcoholic steatosis is a chronic disease characterized by lipid accumulation in the cytoplasm of hepatocytes involving early mitochondrial impairment and late hepatocyte cell death. The voltage‐dependent anion channel (VDAC) is a mitochondrial membrane channel mediating the flux of metabolites, calcium and water across the mitochondrial outer membrane. Upon lethal stress, VDAC can mediate mitochondrial membrane permeabilization (MMP) and cell death. Combination of four experimental models of steatosis (human biopsies, isolated mitochondria from ob/ob obese mice, high fat diet‐fed mice or immortalized cell lines) displayed, in steatotic livers, favored calcium‐induced MMP induction and permeability of VDAC. Thus, lipid accumulation increased the influx of water and calcium into mitochondria and sensitized the organelle to matrix swelling, depolarization and cytochrome c release. Moreover, NADH oxidase activity and channel properties of VDAC are sensitized to calcium regulation in steatosis models. These findings are associated with the hypophosphorylation of VDAC on a threonine residue and the loss of its interaction with the anti‐apoptotic Bcl‐XL and GSK3 kinase. In conclusion, VDAC acts as an early sensor of lipid toxicity and its GSK3‐mediated phosphorylation status controls outer mitochondrial membrane permeabilization in steatosis.
Changes in expression of Multiple Inositol Polyphosphate Phosphatase1 (Minpp1) under Apoptotic and Cellular Stress Conditions
Authors: Kilaparty Surya Pramila , Ali Nawab
Published: June 2021

Inositol pholyphosphates (InsPs) are ubiquitous in nature. They play crucial roles in diverse cellular processes. Multiple Inositol Polyphosphate Phosphatase (Minpp1) is an ER luminal protein implicated in dephosphorylation of InsPs in vitro. However, its physiological function in vivo is unclear. Recent studies have shown that it also dephosphorylates compounds other than InsPs and its expression is altered during chondrocyte differentiation. ER resident proteins involve in protein folding, maturation and transport to various cellular destinations and are affected by external ER stressors. ER stress is likely to influence cellular processes regulated by ER proteins including Minpp1. Our previous studies indicated an increased expression of Minpp1 during apoptosis. Therefore, in order to understand physiological relevance of Minpp1 in ER related functions, in this study, we have compared changes in its expression due to apoptosis with other stress conditions. Using Western blot analysis in a mouse osteoblastic MC3T3 cell line, we found an increased expression of Minpp1 upon induction of heat shock, osmotic and oxidative stresses. Our results suggest that Minpp1 may play a role in the regulation of ER stress and in apoptosis. Physiological relevance of the altered expression of Minpp1 is being further explored following gene silencing with Minpp1 siRNA.
Development of a Novel Reporter System to Monitor ER stress
Authors: Moon Ja Young , Back Sung Hoon , Han Jaeseok , Kaufman Randal
Published: June 2021

A new reporter system to monitor ER stress by measuring ATF6a activation was developed. For this, four recombinant reporter vectors were constructed by using two promoters, pCMV or pTK, followed by coding sequences of Gal4 DNA‐binding domain (BD) andVP16 activation domain (AD) and C‐terminal part of human ATF6a (aa362–670 or aa374–670). As expected, the GAL4 DB‐VP16 AD‐hATF6a fusion protein expressed from the recombinant expression vectors was localized in the ER. Western blot analysis confirmed that both GAL4 DB‐VP16 AD‐hATF6a (aa362–670 or aa374–670) fusion proteins were cleaved to GAL4‐VP16 fragment and hATF6a (aa362–670 or aa374–670) proteins by ER stress. The PathDetect HeLa Luciferase Reporter (HLR) cell line stably established with pCMV‐GAL4‐VP16‐hATF6a(aa374–670) expression vector showed 7‐fold induction of dual luciferase activity by DTT treatment. In conclusion, the HLR‐GVATF6 cell line could be used as a novel reporter system to monitor ER stress by measuring luciferase activity induced by GAL4‐DB and VP16‐AD chimeric transcription factor during ER stress.
Biological impact of as‐produced and purified Single Wall Carbon Nanotube in Kidney epithelial cells.
Authors: Goornavar Virupaxi , Biradar Santoshkumar , Pradhan Aswini K. , Ramesh Govindarajan T.
Published: June 2021

Single Wall carbon nanotubes (SWCNT), due to their unique and outstanding properties, have generated extraordinary interest and expectations for industrial and biomedical applications. The increasing mass production of SWCNT requires their biocompatibility studies to prevent the possible health hazards among workers involved in the research, manufacturing of these materials and in the general public. The as‐produced and purified SWCNT were characterized by FESEM, TEM, FTIR and the purity was checked by TGA and EDS. Kidney epithelial cell lines were used to investigate the biological impact induced by as‐produced and purified SWCNT. Our results showed a significant amount of incorporation of dUTPs in the nucleus in the as‐produced single wall carbon nanotube confirming the induction of apoptosis, which was also validated by DNA fragmentation assay. The activity of caspases‐3 and caspase‐8 in as‐produced SWCNT increased in a dose and time dependent manner in comparison with the purified SWCNT parallel to the morphological apoptosis. Also the decrease in the antioxidants levels like SOD, GSH was in a higher ratio with the as‐produced SWCNT compared to purified SWCNT. Together, we present a complete evaluation of stress levels induced, antioxidants depletion, activation of caspase cascade by as‐produced and purified SWCNTs.
Elucidating the role of apoptosis during cyclical body regeneration in Botryllus schlosseri
Authors: Adamo Meredith Ann , Lauzon Robert
Published: June 2021

Apoptosis is the principal form of programmed cell death by which multicellular animals rid themselves of old cells in order to allow new cells to grow. The colonial ascidian Botryllus schlosseri is a model system for homeostatic cell death and regeneration. During its unique, weekly budding cycle known as blastogenesis, new buds are produced asexually and eventually replace the old adult zooid generation through resorption via circulating phagocytes. Using polyester wax‐embedded tissue sections, we have recently shown that TUNEL‐positive (terminal deoxynucleotidyl transferase nick end‐labeling), apoptotic cells were observed within circulating phagocytes during the colony's death phase (takeover). In contrast, the cells in regressing organs and tissues that had not yet been phagocytosed were TUNEL‐negative. The goal of the current study is to determine whether cells from regressing adults undergo internucleosomal DNA fragmentation characteristic of apoptosis or whether they utilize an alternative cell death mechanism not involving DNA fragmentation, such that they are undetected using TUNEL labeling. Using serial tissue sections from colonies isolated at various stages of takeover as well as budectomized colonies during which adults die but exhibit curtailed resorption, work is currently underway to address these questions to obtain a more holistic understanding of B. schlosseri blastogenesis.
Soluble ST2 controls allergic inflammation in a murine model of asthma
Authors: Hayakawa Hiroko , Hayakawa Morisada , Ohto‐Ozaki Hiromi , Tominaga Shin‐ichi
Published: June 2021

Soluble ST2 (sST2) belongs to the interleukin (IL)‐1 receptor family and has only extracellular domain of ST2L. IL‐33 induces the production of Th2 cytokines via ST2L and causes allergic diseases. On the other hand, sST2 binds IL‐33 as a decoy receptor and suppresses the IL‐33/ST2L signaling. Thus, sST2 may play important role in the allergic immune response and pathogenesis of IL‐33‐related diseases. To explore the role of sST2 in these processes in vivo, we created transgenic mice that constitutively express murine sST2 cDNA (sST2‐Tg mice). sST2‐Tg mice were without any apparent abnormalities in development and growth. In lymphoid tissues of the mice, total cell numbers and population of T and B cells were also normal. Analysis using ST2L‐expressing cells showed that serum derived from sST2‐Tg mice suppressed the activation of MAP kinases and NF‐kappaB in the IL‐33/ST2L signaling. In this context, sST2‐Tg mice were subjected to the ovalbumin‐induced asthma model, resulting in marked reduction of lung inflammation compared to that of littermates. Analysis of lung revealed the reduction of eosinophil infiltration and IL‐4, IL‐5, and IL‐13 production. These results suggest that sST2 suppresses the development of lung inflammation in allergic asthma. Therefore, sST2 is a potential therapeutic agent in allergic diseases. This study was supported by the grant of Takeda Science Foundation (to Morisada Hayakawa).
Suppression of Scopolamine‐induced Cognitive Impairment by Dehydroglyasperin C
Authors: Kim Jong‐Sang , Seo Ji Yeon , Park In Sil , Kim Seong Soon , Kang Hye Ryung , Kim Bo Ram , Zakharova Alexandra , Kim Hyo Jung
Published: June 2021

Our previous study demonstrated that dehydroglyasperin C (DGC) isolated from licorice had neuroprotective effects in mouse hippocampal cells. This study was conducted to assess the protective effect of DGC on the impairment of memory and cognitive function in scopolamine‐treated mice. ICR mice (5 week‐old, male) were injected i.p. with 1 mg/kg body weight of scopolamine to induce amnesia. The ability of learning memory of mice was evaluated by passive avoidance test. The spatial learning and memory abilities of mice were assessed by Y‐maze test and Morris water maze test. We found that DGC improved the cognitive function of mice injected with scopolamine in a dose‐dependent manner. In particular, the group treated with 10 mg DGC/kg body weight showed the longer latency time than the control group. Moreover, DGC suppressed the scopolamine‐induced spatial memory impairment as assessed by Y‐maze test. The inhibitory activity of DGC against acetylcholine esterase present in hippocampus and cortex extracts was also observed. In conclusion, these results suggest that DGC attenuates scopolamine‐induced memory impairment maybe through enhancement of the cholinergic nervous system via acetylcholinesterase inhibition.

Grant Funding Source: National Research Foundation of Korea
Adequacy of the intake of vitamins and mineral in women participants in the Mexican National Nutrition Survey 2006 (ENSANUT‐2006)
Authors: Villalpando Salvador , Ramirez‐Silva Ivonne , Moreno‐Saracho Jessica , Shamah‐Levy Teresa , Beltran‐Velázquez Filiberto , Campos‐Vega Rocio
Published: June 2021

Objective

To evaluate the adequacy of intake of micronutrients and fiber in women from the ENSANUT‐2006.

Methods

Dietary intake and compliance with dietary recommendations were estimates for a sample of Mexican women aged >;19 years from ENSANUT‐2006. Food intake came from a food frequency questionnaire. Dietary adequacy was classified into adequate (adequacy >;100%), moderately inadequate (50–99%), and highly inadequate (<50%).

Analysis

Means of adequacy were estimated through linear and polynomial multiple regression.

Results

The adequacy for vitamin A varied by age from 69–79%, the following intakes decreased progressively with age: Vitamin C 69–50%, folate 26–12.4%, vitamin B12 56.4–34.4%. For minerals iron decreased from 21.2–12‐13% and increased to 45% in >;50 years of age. Calcium increased with age from 24.9–32.9% and declined to 15.0%. Magnesium intake decreased from 56.6–36.9% and phosphate increased from 57.8–88.2%. The adequacy for fiber increased with age from 12–36%.

Discussion

We documented evidence about the low dietary intake of vitamins and minerals prevailing in Mexican women, although the coincidence with the prevalence of micronutrients deficiencies only occurred with vitamins A and C, calcium, magnesium.
The influence of “Guiding Stars” on perception and choice of foods at a 4‐year university
Authors: Carey Gale Beliveau , Maher Michelle , Laramore Riley , Pascoe Caitlin , Skalinski Jessica , Morrell Jesse
Published: June 2021

“Guiding Stars” ™ (GS) is a point of purchase, consumer tool developed in 2006 that uses a nutrient density algorithm to categorize foods as good (one star), better (2 stars) or best (3 stars). GS was originally designed for grocery stores. The University of New Hampshire has adopted GS as a nutrition guidance tool for its students. The goal of this study was to determine if the presence of GS in the Union Court (UC) influenced 18 to 24 year old students’ perception and choice of foods. A 4‐question survey was devised to determine demographics, nutrition education background, ease of identifying healthy food choices, and factors influencing food selections, of UC patrons. The survey was conducted in November 2011 (pre‐GS, n=194) and after GS was introduced to the UC in April 2012 (post‐GS, n=199). Survey results revealed that students did not perceive that GS influenced their food selections, but did perceive that healthy food choices were significantly more available after GS (49%) than before GS (34%). After GS, students significantly decreased their perception that healthy foods were rarely/never available, from 22% to 8%, compared to before GS. Regardless of the presence of GS, a prior nutrition course was found to significantly increase, by 2‐fold, student awareness of the nutrient content/health of a food and posted nutrition/calorie information. Although students report that GS did not influence their food choices, GS did increase student perception that healthy foods were readily available at the UC. Whether this perception will translate into changes in food choices, remains to be determined. Supported by NHAES grant NH00541
Major food sources of heavy metals differ between the high and low dietary exposure groups
Authors: Oh Se‐Young , Kwon Sung‐Ok , Hwang Hyojung , Kim Jeongseon , Lee Sang‐Ah , Park Kyung Soo , Kwon Hojang
Published: June 2021

Intakes of vegetables, fruits and grains have been suggested as major contributors of heavy metals such as lead (Pb) and cadmium (Cd), yet these foods are known to be relevant to better health and nutrition. This study estimated major food sources of heavy metals in the high (90–99%) and low (1–10%) exposure groups, as part of the nationwide study with 2,105 adults in Korea. Top three contributing foods of dietary Pb exposure were kimchi, rice and scallion (16.3% for all three) for the low dietary Pb group and watermelon (22.5%), mussel and mudfish (35.9% for all three) for the high Pb group. White rice (35%), kimchi and noodles (45.9% for all three) were major food sources of dietary Cd in the low Cd group while squid (36.7%) and rice (11.8%) consisted of 48.6% of Cd exposure in the high dietary Cd group. Cabbages provided the highest proportion (30%) of dietary Hg in the low Hg group and white fishes and mackerel (40% for all two) were important sources in the high Hg group. These findings suggest that fish and shellfish intakes may play a significant role in dietary exposures of Pb and Cd in addition to Hg. (Supported by grant No. S‐12–04‐2‐ PMP‐994–0‐F from the Korea Food & Drug Administration)
State‐wide availability of pulse crops in USDA Foods Program
Authors: Waldrop Megan E. , Rueda Janice M.W.
Published: June 2021

Childhood obesity remains a critical public health concern, and school lunches provide a key entry point for more nutritious foods into the diets of school children. The USDA Foods Program is an important source of affordable American grown foods for schools participating in the National School Lunch Program (NSLP). Pulse crops, which include dry beans, dry peas, lentils and chickpeas, are among the foods available as part of the USDA Foods program. The aim of this study was to describe the statewide availability of the 16 different pulse foods included in this program. Requests were e‐mailed to the NSLP coordinators in all 50 states, and a 94% response rate was achieved. Results show that not all states offer the complete selection of pulse crops available as part of the USDA Foods Program. Frequency of availability of each item was analyzed, and regional differences in availability were determined.
Cooking Show and Tell: An educational tool for improving vegetable preference among first graders
Authors: Colas Marie V , Vaccaro Joan A , Zarini Gustavo G , Huffman Fatma G
Published: June 2021

A pre (raw vegetable) and post (Blanched/Sautéed) cooking show and tell demonstration was conducted. Children of ages 6 and 7 were recruited from two first grade classes; 52 students 18 boys and 34 girls had parental consent and participated. A parent questionnaire was sent home to evaluate parental influences with respect to vegetable liking. Paired samples t‐test revealed a significant difference in preference of raw vegetables (p < .001) and no gender differences for the preference of raw compared to cooked vegetables (p < .005). A McNemar‐Bowker test revealed a significant difference in vegetable preference for cauliflower, bell pepper, carrots, green beans, celery. Parental influences were associated with vegetable intake for certain vegetables. Children who are exposed to a cooking demonstration and nutrition education intervention tend to be receptive and become open to trying new foods. There were gender differences in preferences for raw vegetable. Further research is needed to evaluate the show and tell intervention method for different populations and over a longer period.
Long‐term effects of micronutrient supplementation on school age child behavior
Authors: Bahnfleth C–arlotte Lena , Murray‐Kolb Laura E , Schaefer Barbara , Cole Pamela , Katz Joanne , Khatry Subarna K , LeClerq Steven , Tielsch James M , Christian Parul
Published: June 2021

Data on long‐term effects of preschool micronutrient supplementation on behavior are sparse. We examined behavior in 7–9 y old Nepali children (n=694) who received micronutrient supplements from 12–35 mo of age in a double‐masked RCT. Children were randomized to daily iron‐folic acid (IFA); zinc (Zn); iron‐folic acid+zinc (IFAZn); placebo (PL). Behavior was assessed on six dimensions (positive mood; negative mood; lively/active; sociability; sustained attention; demandingness) during two visits. Differences in behavioral ratings were examined relative to the PL for each supplement group. Additionally, those who received any zinc (Zn or IFAZn) or iron (IFA or IFAZn) were compared to those who did not. Overall, those who received IFA scored higher in sustained attention (p=0.05) and demandingness (p=0.02) and those who received IFAZn scored higher in sociability (p=0.04) compared to PL. After adjusting for confounders, the only difference remaining was higher demandingness scores in those who received IFA compared to PL (p=0.002). In both unadjusted and adjusted analyses, children who had received zinc vs. not were rated significantly higher in sociability (p=0.0007 and p=0.009, respectively), but no differences were found between those who received iron vs. not. Zinc supplementation between 12–35 mo appears to have a positive effect on sociability at 7–9 y.

Grant Funding Source: NICHD and Gates Foundation
Iron deficiency but not anemia affects cognition in Indian adolescents
Authors: Scott Samuel , Murray‐Kolb Laura , Wenger Michael , Udipi Shobha , Ghugre Padmini , Haas Jere
Published: June 2021

A quarter of the world's population is thought to be deficient in iron. Though the functional effects of iron deficiency have been studied in infancy and adulthood, adolescence, a period of significant neural and behavioral maturation, has not received much attention. In this cross‐sectional analysis of Indian adolescents in rural Maharashtra (n = 147; 12–16 y; 61 females), relations between baseline blood variables and performance on a test of simple reaction time (SRT, in ms, one of a set of five measures of perceptual and cognitive functioning) were examined. Iron status was defined according to stage of iron deficiency (iron‐deficient anemic (IDA), iron‐deficient (ID), iron sufficient (IS)). Those in the IDA and ID groups had longer RT than those in the IS group (p=0.005 & 0.013, respectively). SRT for individuals who were anemic but not ID did not differ from those of the IS group but were faster than those in the IDA and ID groups (both p<0.01). Further analyses revealed that the serum ferritin quartile was strongly related to SRT (p<0.001) whereas hemoglobin quartile was not. Finally, those with positive body iron (calculated by Cook's equation) had shorter SRTs than those with negative body iron (p=0.044). We conclude that iron deficiency with or without anemia is associated with slower responding, whereas anemia in the absence of iron deficiency is unrelated to response speed in Indian adolescents.

Grant Funding Source: HarvestPlus
Prospective, Randomized Double Blind Study to Test Vitamin A Treatment in Patients with Sepsis
Authors: Tayek John A
Published: June 2021

Purpose

To test the benefits of Vitamin A Tx patients with sepsis on length of time in ICU, Days on Ventilator, Days on Pressors and 28‐day mortality. The trial was prospective, randomized and double‐blind. As part of a larger sepsis trial, 63 patients with sepsis were randomized to receive either 50,000 IU of Vitamin A intramuscular or placebo over 7‐days. Data analysis was by ANOVA with two tailed test and p<0.05 as significant.

Results

The mean age was 51+2 (mean+ SEM) with 54% female. All patients had an ACTH stimulation test using 250 mcg of ACTH IV and serum cortisol was measured at time 0, 30 and 60 minutes. Baseline cortisol of 24.6+1 mg/dl increased to 41+2 mg/dl at 30 minutes and 49+2 at 60 minutes. There was no significant difference between the groups. All cortisol responses were greater than 12.9 mg/dl. Serum Vitamin A level was below normal in 54% of the patients tested (n=35). Table 1 describes the two groups with regard to baseline APACHE III score, baseline WBC and percent of patients with bacteremia. Vitamin A was given to 32 patients and placebo was given to 31 patients.

Conclusions

The number of days in the ICU was slightly, but not significantly (p=0.13), reduced in Vitamin A treated patients. The average number of days on pressors, the number of days on the ventilator and the 28‐day mortality rate were also not significantly different. Vitamin A Tx in septic adult patients failed to significantly reduce the number of days in the ICU, days on the ventilator, days on pressor agents or 28‐day mortality. Seven days of high dose Vitamin A Tx has no benefit in adults with sepsis (mean+sem).
Tree nut consumption is associated with better diet quality, nutrient intake of select nutrients, and better measures of some cardiovascular risk factors (CVRF): National Health and Nutrition Examination Survey (NHANES) 2005–2010
Authors: O'Neil Carol E. , Nicklas Theresa A , Fulgoni Victor L
Published: June 2021

NHANES data (2005–2010) were used to assess the association of tree nut consumption by adults (N=14,386) with diet quality, select nutrient intake, and CVRF factors. Covariate adjusted least square means ± SE were determined. Diet quality was determined by the Healthy Eating Index (HEI) score. HEI was higher in tree nut consumers than in non consumers (60±0.6 v 51±0.3; p<0.0001). Tree nut consumers had higher (p<0.0001) daily intakes of energy (2468±36 v 2127±13 kcal) and nutrients of concern: fiber (20.7±0.6 v 15.8±0.2 g); potassium (3028±51 v 2691±14 mg); magnesium (408±9 v 292±1 mg); MUFA (35.5±0.5 v 29.3±0.1 g), and PUFA (21.0±0.5 v 17.2±0.1 g), but lower intakes of added sugars (14.7±0.6 v 18.1±0.3 tsp), SFA (25.2±0.4 v 27.2±0.1 g), and sodium (3197±49 v 3570±13 mg) than non‐consumers. Tree nut consumers had lower weight (79.9±0.8 v 82.3±0.3 kg; p=0.0049), BMI (27.7±0.3 v 28.7±0.1; p<0.0001), and waist circumference (95.7±0.7 v 98.1±0.3 cm; p=0.0006) than nonconsumers. Tree nut consumers also had lower systolic blood pressure (120.4±0.7 v 122.1±0.2 mmHg; p=0.0120), higher HDL‐cholesterol (54.8±0.6 v 52.9±0.2 mg/dL; p=0.0020), and lower HOMA‐IR (3.0±0.1 v 3.3±0.1; p=0.0498). Consumption of tree nuts should be encouraged to improve diet quality, nutrient intake, weight status, and some CVRF. Support: USDA & International Tree Nut Council Nutrition Research & Education Foundation
What role does moderate alcoholic beverage intake play in cardiovascular health?
Authors: Pourafshar Shirin , Johnson Sarah A. , Jafarinasabian Pegah , Bonn Mark , Arjmandi Bahram H.
Published: June 2021

Cardiovascular disease is the number one cause of death in the United States. Red wine consumption has been promoted for its positive effects on high‐density lipoprotein cholesterol; however, the effects of moderate and heavy alcoholic beverage consumption on cardiovascular health remain unclear. The objective of this cross‐sectional study is to investigate the role of alcoholic beverage intake on cardiovascular health. A total of 60 men and women aged 25 to 40 years old with a body mass index (BMI) between 18.5 and 35 will be included. Participants will be divided into seven quartiles (never or less than 1 drink per week, 1 drink per week, 2–3 drinks per week, 4–6 drinks per week, 1 drink per day, 2+ drinks per day, and 3+ drinks per day) based on their alcoholic beverage intake as indicated in the Beverage Questionnaires. Overnight fasting blood measurements will include C‐reactive protein and lipid profiles. Atherogenic risk ratios, waist‐to‐hip ratio, and BMI will be calculated. Body composition will be assessed using dual‐energy X‐ray. Seated blood pressure (BP) will be assessed by measuring resting brachial BP. Pearson correlation coefficients will be used to determine the relationship between variables of interest. The findings of this study will further our understanding of the role of alcoholic beverage consumption in cardiovascular health.

Grant Funding Source: Dedman School of Hospitality
Yogurt consumption by US children is associated with higher protein, calcium and vitamin D intake, lower dietary fat, and lower body fat: Results from the National Health and Nutrition Examination Survey (NHANES 2005–2008)
Authors: Keast Debra R , Albertson Ann M , Gugger Carolyn K , Holschuh Norton M
Published: June 2021

This study examined the associations of dairy and yogurt consumption with intakes of macronutrients, calcium (CA), and vitamin D (VIT D), and obesity measures among American children. Using 1‐day dietary recall data from NHANES 2005–2008, children aged 8–18 yr (n=3,786) were classified by dairy consumption: <1 (31%), 1‐<2 (28%), or 2+ servings (41%). Yogurt consumers (Y‐C) (8.5%) and nonconsumers (Y‐NC) (91.5%) were defined using 2‐day intake. After adjusting for gender, ethnicity, age, and energy (Model 1), dairy consumption was associated with higher intakes of protein, CA, and VIT D (p<0.01); and Y‐C had higher intakes than Y‐NC (p<0.01). Dairy consumption was also associated with higher energy and saturated fat intakes, although carbohydrate and added sugars were lower. Y‐C had lower intakes than Y‐NC of total fat (31.6±0.7 vs. 33.5±0.3 % energy) and saturated fat (10.9±0.3 vs. 11.7±0.1 % energy; 26.4±0.8 vs. 28.2±0.3 g), with no differences in energy, carbohydrate, total sugars, and added sugars. With physical activity and other covariates added to Model 1, subscapular skinfold was lower for Y‐C than Y‐NC (11.3±0.6 vs. 12.9±0.3 mm), and for consumers of 2+ vs. <1 dairy servings (12.3±0.2 vs. 13.4±0.3 mm). Y‐C also had lower waist circumference (74.5±1.0 vs. 77.0±0.5 cm) than Y‐NC. Yogurt consumption was associated with higher protein, CA and VIT D intake, lower dietary fat and body fat, with no differences in sugars intake. Supported by the Bell Institute of Health and Nutrition, General Mills, Inc.