bluuebunny/crossref_metadata_2025 huggingface dataset:
Striking and Preferential Expression of ADRM1 (adhesion regulating molecule1) in Colon, and Lung Cancers
Authors: Cha Mee‐Kyung , Kim Il‐Han
Published: June 2021

The proteasome is a multiprotein complex that regulates the stability of hundreds of cellular proteins and thus, it is implicated in virtually all cellular functions such as the regulation of cell proliferation or cell cycle control, transcriptional regulation, immune and stress response, cell differentiation, and apoptosis. ADRM1 (adhesion regulating molecule1) is a known component of the proteasome. As for some other proteasome subunits, its exact role is not clearly known. ADRM1 is abundant and widely distributed both phylogenetically and in tissues. We investigated levels of expression of ADRM1 gene in various cancer tissues by real‐time polymerase chain reaction. Levels of ADRM1messenger RNA (mRNA) in colon and lung cancer tissues were much higher than those in other solid cancers. ADRM1 gene was significantly induced in colon, lung, breast, and ovary cancers, whereas did not significantly in thyroid, liver, prostate, and kidney cancers. Western blot analysis of the various cancer tissues revealed consistent and preferential expression of ADRM1 in colon and lung cancers. Taken together, the striking and preferential inductions of ADRM1 in colon and lung cancers may enable their use as corresponding cancer markers. *This work was financially supported by Regional Research and Development Cluster Project (B0009735) funded by the Ministry of Knowledge Economy (MKE) of Korea.
Determination of the intracellular aqueous/lipid interface of integrin alpha 2b in vivo
Authors: Zhu Quansheng , Kao Liyo , Liu Weixin , Newman Debra , Kurtz Ira
Published: June 2021

Integrins are a family of cell adhesion receptors that mediate cell migration, extracellular matrix assembly and platelet aggregation. They are heterodimeric proteins composed of alpha and beta subunits. Integrins exist in a default low affinity state in the cell membrane. The highly conserved intracellular memebrane proximal region of the alpha subunit is considered to play a critical role in constraining the receptor in the low affinity state, however, the location of this region in the cellular environment remains controversial. In this study, we explored the location of the membrane proximal region of integrin alpha 2b by substituted cysteine accessibility method (SCAM). Residues in the membrane proximal region of integrin alpha 2b were individually substituted with cysteines and subjected to in situ chemical probing with a cysteine specific reagent, biotin maleimide. Results showed that Lys‐994 marks the first aqueous exposed residue in the region, consistent with the in vitro findings. Further functional analysis with a flow cytometry assay revealed that all the mutant receptors were active. On the basis of the results, we conclude that the in vitro determined aqueous/lipid interface of integrin alpha 2b represents the active state of the receptor.

Source of Research Support: UCLA Nephrology Research Fund
Technology Development for Studying the Dynamics of Protein‐protein Associations on Chromatin
Authors: Tackett Alan
Published: June 2021

We are optimizing technologies for quantitative analysis of proteins and histone post‐translational modifications associated with distinct chromatin loci. Key to employing these approaches is the sufficient use of chemical cross‐linking to minimize protein exchange during chromatin isolation; conversely, over cross‐linking renders chromatin insoluble. We developed a quantitative method termed transient I‐DIRT, useful for determining optimal levels of chromatin cross‐linking. We have applied transient I‐DIRT to define the local interactome of a low‐abundance yeast histone acetyltransferase (NuA3), and have also applied this method to determine cross‐linking conditions for efficient chromatin purification with minimal histone exchange. Our results provide insight into the dynamics of protein‐protein association on chromatin and establish the parameters for purification of native chromatin sections.
Extending XMLPipeDB with GO Slim to Update the GenMAPP‐compatible Gene Database for Budding Yeast, Saccharomyces cerevisiae, for the Analysis of DNA Microarray Data
Authors: Pak Bernadette , Murphy Don , Dionisio John David N. , Dahlquist Kam D
Published: June 2021

The GenMAPP suite is a powerful tool for analyzing DNA microarray data on biological pathways and performing Gene Ontology term over‐representation analysis. However, the use of GenMAPP and MAPPFinder is limited to those species for which a GenMAPP‐compatible Gene Database exists. Furthermore, the process of creating a Gene Database is sensitive to changes in flat file formats. To simplify database updating and maintenance, and to be robust to changes in file formats, XMLPipeDB, an open source tool chain for building relational databases from XML sources, was created. XMLPipeDB provides functionality for database configuration, importing data, performing queries, and exporting GenMAPP‐compatible Gene Databases based on data from UniProt and Gene Ontology (GO). In order to take advantage of new gene annotations and update the 2006 Gene Database for Saccharomyces cerevisiae, we extended XMLPipeDB by integrating GO term associations with the XML‐derived S. cerevisiae data from UniProt and GO and implementing GO Slim, a scaled‐down version of the Gene Ontology that overcomes memory limitations in the MAPPFinder software. Once we updated the database, we used it to analyze DNA microarray data from Arava et al. (2003) to determine which genes in yeast are regulated by translational control. We found that mRNAs that encode proteins involved in the GO process tRNA aminoacylation were over‐represented among the pool of mRNA being actively translated, a result not previously reported.
Detection of differentially expressed Beta Amyloid fragments from Alzheimer's Patients by Mass Spectrometry
Authors: Roth Steven Howard , Bulman Amanda , Thulasiraman Vanitha , Berardini Mark , Rusa Mariana
Published: June 2021

Study Objective

A MALDI surface with covalently immobilized antibody specific to n‐terminus Beta Amyloid fragments was used to enrich specific fragments from biological samples for detection on a Bruker MALDI TOF/TOF instrument. Comparison of healthy and Alzheimer's disease diagnosed samples was evaluated by this method to determine if Beta Amyloid fragments are differentially expressed between the groups.

Methods

Human CSF pools from diagnosed Alzheimer's disease patients and healthy donors were treated with 9 M urea, 2% CHAPS, diluted in PBS, and applied in specific volume to a 2 mm antibody coated spot demarcated by a hydrophobic barrier. A similar treatment was used to prepare human brain cortex lysates from post mortem samples of Alzheimer's disease diagnosed and non‐Alzheimer's disease individuals. In a third set, brain samples from APP transgenic and wild type mice were prepared in the same fashion. After allowing samples to interact with the antibody surface, washes were done using PBS, PBS with Triton 100X, and 20 mM HEPES. Alpha‐cyano‐4‐hydroxycinnamic acid was applied to each spot after drying. All samples were read in linear mode in a MALDI TOF/TOF instrument.

Results

In each set of biological sample types evaluated, the Alzheimer's diagnosed group was found to express Beta Amyloid fragments, while none were detected in any of the control groups.
Characterization of the SMTNL1‐tropomyosin binding interface using hydrogen‐deuterium exchange mass spectrometry
Authors: Butler Eric Ian , Ulke‐Lemée Annegret , Ishida Hiroaki , Vogel Hans , Chik John , MacDonald Justin
Published: June 2021

The smoothelin‐like 1 protein (SMTNL1) modulates muscle contractile activity, particularly in exercise adaptation. SMTNL1 bears sequence similarity to smoothelin, a smooth muscle differentiation marker, particularly in its calponin homology (CH) domain. The interaction of SMTNL1 with tropomyosin (TM) is dependent on the CH domain and some portion of a central, unfolded region (aa 197–342) of SMTNL1. These data are based on pull‐down studies with SMTNL1 truncations, and reinforced by isothermal titration calorimetry experiments. We hypothesize that independent sections of the SMTNL1 protein cooperate to form a functional TM binding surface. We will elucidate the specific surfaces of SMTNL1 involved in TM binding using hydrogen‐deuterium exchange mass spectroscopy (HXMS). To this end, we have mapped pepsin‐derived mass fragments covering 94% of the SMTNL1‐CH domain, and 83% of a functional TM‐binding truncate (SMTNL1‐∆N, aa 196–459). These maps will be compared with deuterated SMTNL1 samples, and with SMTNL1 samples deuterated while bound to TM. The deuterium incorporated is calculated for each SMTNL1 peptide to reveal the interaction interface. The objectives will address the molecular and structural elements of the SMTNL1‐TM interaction and provide a framework for the physiological role of SMTNL1 in vascular smooth muscle biology. The research was supported by HSFC and CIHR.
Phospholiase C‐eta1 is activated by intracellular Ca2+ mobilization and enhances GPCRs‐mediated signaling
Authors: Kim Jung Kuk , Choi Jung Woong , Lim Seyoung , Ryu Sung Ho , Suh Pann‐Ghill
Published: June 2021

Phospholipase C‐η1 (PLC‐η1) is the most recently identified PLC isotype and mainly expressed in nerve tissue. In the present study, we first report that PLC‐η1 acts as a signal amplifier in G protein‐coupled receptors (GPCRs)‐mediated PLC/Ca2+ signaling in neuronal cell. RNA interference‐mediated knockdown of endogenous PLC‐η1 reduced LPA, BK, and PACAP‐induced PLC activity in mouse neuroblastoma Neuro2A (N2A) cells, indicating that activation of PLC‐η1 is regulated by GPCRs‐mediated signaling. Interestingly, ionomycin‐induced PLC activity was significantly decreased in PLC‐η1 knockdown, but not in PLC‐η2 knockdown N2A cells. In addition, the IP3 receptor blocker, 2‐APB was found to inhibit LPA‐induced PLC activity in control, but it had no more inhibitory effect in PLC‐η1 knockdown N2A cells, suggesting the pivotal role of intracellular Ca2+mobilization in PLC‐η1 activation. Moreover, LPA‐induced ERK1/2 phosphorylation and downstream target gene, krox‐24, expression was significantly decreased by PLC‐η1 knockdown. Finally, we found that 2‐APB inhibited both ERK1/2 phosphorylation and krox‐24 expression only in control, but it had no more inhibitory effect in PLC‐η1 knockdown N2A cells. Taken together, our results strongly suggest that PLC‐η1 is secondarily activated via Ca2+ mobilized from ER and therefore amplifies GPCRs‐mediated signaling in N2A cells. This study was supported by the National Research Foundation of Korea Grant funded by the Korean Government (KRF‐2007‐341‐C00027)
Molecular Dissection of Receptor Tyrosine Kinase Regulation by Cbl Proteins
Authors: Ahmad Gulzar , Reddi Alagarsamy L , Naramura Mayumi , Band Hamid
Published: June 2021

Aberrations of receptor tyrosine kinases (RTKs) have been implicated in the etiology of human cancers. Members of the Casitas B‐lineage Lymphoma (Cbl) family of ubiquitin ligases play a role as negative regulators of RTK function and mutations of Cbl proteins are involved in human cancers such as leukemia. The purpose of studies here is to determine the molecular mechanisms of oncogenic signaling and to explore possible defects in negative regulation of RTK signaling by Cbl proteins. We hypothesize that Cbl and Cbl‐b represent redundant yet essential components to facilitate lysosomal degradation of RTKs. In initial studies, we have derived wildtype, Cbl‐null, Cbl‐b‐null and cbl/cbl‐b‐null mouse embryonic fibroblast (MEF) cell lines to demonstrate an important role of both Cbl and Cbl‐b proteins in RTK ubiquitination and degradation. Absence of Cbl and Cbl‐b expression dramatically alters actin cytoskeleton reorganization upon attachment to extracellular matrix and markedly enhances growth factor‐dependent cell migration indicating that Cbl proteins provide functionally‐important negative regulatory feedback upon RTK activation. Further studies are underway to express specific RTKs (such as human EGFR) in the characterized MEFs together with reconstitution with wildtype or mutant Cbl proteins to dissect the fundamental steps in endocytic traffic where Cbl proteins play an essential role. These studies will focus on the role of Cbl proteins in initial endocytosis vs. later endosomal sorting steps as the consequences of these mechanisms are distinct. This work was supported by: the NIH grant CA99163 to HB.
Structural and Enzymatic Analysis of Orf6, a Novel Dehydratase from a Deep‐Sea Polyunsaturated Fatty Acid Synthase
Authors: Rodríguez‐Guilbe María M , Schreiter Eric , Baerga Abel
Published: June 2021

The polyketide synthase (PKS) genes responsible for eicosapentanoic acid (EPA) biosynthesis in the deep‐sea bacterium, Photobacterium profundum, have been cloned and sequenced, being a total of five polyunsaturated fatty acid (pfa) genes (pfaA, pfaB, pfaC, pfaD and Orf6) enough for EPA production. One of these enzymes is Orf6, a putative thioesterase (TE) domain believed to perform the last step in the fatty acid biosynthesis, catalyzing the hydrolysis and release of the final product from the carrier protein. The purpose of our research is to make a structure‐function analysis of Orf6 TE using its three dimensional structure determined by X‐ray crystallography to guide the re‐engineering of the enzyme. In parallel, we are also performing in vitro enzymatic assays with its likely substrates loaded onto coenzyme A (CoA). The Orf6 structure determined at 1.0 Å resolution reveals a novel hot‐dog fold containing a conserved amino acid, Asp17, in the active site. Results from activity assays indicate that Orf6 does not hydrolyze the CoA thioesters of long‐chain fatty acids. Additional efforts include in vitro assays using CoA thioesters of short‐chain carboxylic acids or acyl carrier protein (ACP) thioesters as substrates for Orf6. Structural and mechanistic knowledge of this seemingly novel enzyme will be important for the design of lipid‐based drugs or the development of industrial applications like biofuels.
Crystal structure of the Escherichia coli PhnD in complex with 2‐aminoethylphosphonate
Authors: Alicea Ismael , Schreiter Eric R
Published: June 2021

The phnD gene of E. coli putatively encodes the periplasmic binding protein of the phosphonate uptake and utilization pathway. We have crystallized and determined the structure of PhnD to 2.0Å resolution by multi‐wavelength anomalous dispersion methods in complex with the environmentally abundant phosphonate 2‐aminoethylphosphonate (2‐AEP). Similar to other bacterial periplasmic binding proteins, 2‐AEP is bound near the center of mass of PhnD in a cleft formed between two lobes. Extensive hydrogen bonding and electrostatic interactions stabilize the 2‐AEP, which binds to PhnD with an affinity of ~1 nM. Interestingly, the C‐terminus of PhnD comprises two long coiled‐coil alpha helices that mediate dimerization in the crystal structure. This structure provides insight into environmental phosphonate uptake by bacteria and could facilitate the design of phosphonate sensors based on PhnD.
Intramolecular base‐catalysis in the radical‐scavenging reaction by vitamin E derivatives bearing a pyridine moiety
Authors: Nakanishi Ikuo , Yakumaru Haruko , Nyui Minako , Ohkubo Kei , Matsumoto Ken‐ichiro , Fukuhara Kiyoshi , Okuda Haruhiro , Fukuzumi Shunichi , Ozawa Toshihiko , Anzai Kazunori , Ikota Nobuo
Published: June 2021

In order to develop effective radioprotectors, we recently synthesized three vitamin E derivatives bearing a pyridine moiety with a linkage of different spacers. All the vitamin E derivatives efficiently scavenged 2,2‐diphenyl‐1‐picrylhydrazyl radical (DPPH•) in deaerated methanol at 25 °C. The second‐order rate constant for DPPH•‐scavenging by one of the vitamin E derivatives determined by the stopped‐flow technique is about 3‐fold larger than that by 2,2,5,7,8‐pentamethylchroman‐6‐ol (PMC), a vitamin E model. This suggests that the pyridine moiety may accelerate the reaction by stabilizing the radical cation generated in the electron transfer from the vitamin E derivative to DPPH•.
Implementing “Problem‐Based Dissection” in a 100 Week Human Structure Course
Authors: Rennie William Price , Metzger Keith A , Gannon Patrick J
Published: June 2021

At our School of Medicine the disciplines of gross anatomy, embryology, histology, gross and histopathology and clinical imaging are integrated into one longitudinal 100 week course, “Structure”, which includes all components in a continuous fashion. Traditionally, laboratory dissection experience is an important component of structure courses, although concern has been raised regarding the efficiency, expense and curricular time allotted for these activities.

To address these challenges, we focused on the contribution structure makes to the identification, explication and solution of clinical problems. “Problem‐based” projects are assigned to groups of 5 students who work as a team over several weeks to research, explore connections and devise dissection approaches that help to explain multiple aspects of the problem. Upon completion of projects, all are presented in a lab‐based “poster session” format, organized in a manner such that all students both present and learn from presenters during the session. Faculty mentors are responsible for guiding student investigations and for evaluation of both group and individual performance. Peer evaluation is also a part of each student's overall evaluation. The addition of this approach to dissection preserves an important tradition and focuses student effort on gaining tangible, useful skills in the application of structural science to clinical medicine.
Self immunity and resistance mechanisms against trojan horse antibiotic ‐ Microcin C7
Authors: Agarwal Vinayak , Tikhonov Anton , Novikova Maria , Severinov Konstantin , Nair Satish
Published: June 2021

Microcin C7 (McC7) antibiotic is a non‐hydrolyzable analog of aspartate‐adenylate and hence inhibits the aspartyl tRNA synthetase enzyme, causing translation arrest and cell death in susceptible bacteria. The antibiotic producer cell has two self‐immunity mechanisms to protect itself against the action of the antibiotic. The mature drug is acetylated by a specific GNAT‐superfamily acetyltransferase, encoded by gene mccE, which leads loss in bioactivity. The drug can also be hydrolyzed by a specific carboxypeptidase, encoded by gene mccF, to yield inactive products. Structural characterization of these enzymatic steps is required for the development of inhibitors, as these self‐immunity genes can be transferred to susceptible cells and employed for resistance against the drug, once McC7 is applied in a clinical setting. High resolution structures of MccE and MccF in their susbstrate bound forms are presented and implications for inhibitor design are discussed. MccF is the first enzyme characterized for the hydrolytic inactivation of phosphoramadite linkage containing antibiotics, and a high resolution structure would provide means for engineering towards diverse substrate tolerance and subsequent development of a novel biocatalyst.
Expression of SOX3 in the urogenital ridge is associated with XX male sex reversal in mice
Authors: Thomas Paul Quinton , Hughes James , Tan Jacqueline , Rogers Nicholas , Sekido Ryohei , Lovell‐Badge Robin , Sutton Edwina
Published: June 2021

Sox3 is expressed in the progenitor cells of the embryonic CNS and functions as a dosage‐dependent regulator of CNS development in mice and man. To further investigate the role of SOX3, we generated transgenic mice using a Sox3 genomic BAC fragment. Unexpectedly, progeny from one line (termed Sex reversed (Sr)) exhibited a markedly distorted sex ratio in favour of male offspring. Analysis of the reproductive tract in adult sex‐reversed animals indicated that all male‐specific structures were present, indicating complete XX male sex reversal. Transgene expression analysis in Sr embryos demonstrated that Sox3 is ectopically upregulated in the primordium of XX and XY gonads during sex determination due to a position effect. Importantly, morphological, gene expression and co‐transfection studies indicate that SOX3 induces testis differentiation in Sr embryonic gonads by activation of Sox9 in a mechanism analogous to that used by the Y‐linked sex determining gene Sry. Together, these data suggest that SOX3 and SRY proteins are functionally interchangeable in sex determination and provide support for the notion that Sry evolved from Sox3 via a regulatory mutation that led to its de novo expression in the early gonad.
Protein‐protein Interactions and Acyl Carrier Protein Recognition in the Biosynthesis of Omega‐3 Fatty Acids by a Polyketide Synthase
Authors: Trujillo Uldaeliz , Rodríguez‐Guilbe María , Oyola‐Robles Delise , Baerga‐Ortiz Abel , Santos Jennifer
Published: June 2021

Acyl carrier protein (ACP) is a highly conserved protein domain that participates in the biosynthesis of fatty acids and polyketides. Most enzyme complexes contain either one or many of these domains arranged in a modular fashion. However, the polyketide synthase (PKS) complex responsible for the production of omega‐3 fatty acids in deep‐sea bacteria contains a total of five ACP domains in tandem, with no obvious reason as to why this particular arrangement has been widely selected throughout nature. One possible explanation is that different ACP domains have different binding specificities towards the enzyme domains which “service” them. In order to investigate whether ACP domains have specificity toward other enzyme domains, we have expressed ACP domains individually in E. coli and purified them using a combination of nickel chromatography and anion exchange chromatography. Each individual ACP will be assayed for binding to the different purified enzyme domains, such as the thioesterase, Orf6, and the dehydratase domains and well as other PKS domains. ACP domains have been expressed with and without the phosphopantetheine modification by co‐expression with a phosphopantetheine transferase. The results from this work will facilitate the mechanistic understanding of marine biosynthetic machineries and will allow the harnessing of genes of marine origin for the production of new natural product with therapeutic potential.

This work was founded by the MBRS‐RISE Program of the University of Puerto Rico Medical Sciences Campus, Grant R25GM061838.
Expression profiling of ETS and MMP factors in VEGF‐activated endothelial cells
Authors: Heo Sun‐Hee , Choi Young‐Jin , Ryoo Hyun‐Mo , Cho Je‐Yoel
Published: June 2021

In the process of angiogenesis, working of many transcription factors at the proper time is important to activate angiogenesis‐related genes. In this study, we searched for Ets transcription factor members and matrix metalloproteinases (MMPs) that respond to VEGF in endothelial cells. We first analyzed the expression of 27 human Ets factors and 15 human MMPs in VEGF‐treated human umbilical vein endothelial cells (HUVEC) using quantitative RT‐PCR. We found that ETV‐1, Fli‐1, ERG, MMP‐1, ‐3, ‐7, ‐8, ‐9, ‐10, ‐13, and ‐19 expression is up‐regulated more than 1.5‐fold in HUVEC after 2 hr of VEGF treatment. In addition, the expression of MMP‐10 induced by VEGF remained 2‐fold higher for 24 hr compared to non‐treated control. The elevation of MMP10 mRNA and protein levels was confirmed to be both time‐ and dosage‐dependent. In addition, MMP‐10 transcription was mediated by Ets‐1. The inhibition of PI3K and MAPK inhibited VEGF‐induced MMP‐10 expression. Furthermore, transfection of MMP‐10 siRNA inhibited VEGF‐induced migration and tube formation, and it also inhibited vessel formation in matrigel plugs in vivo. In conclusion, our study demonstrated induction of MMP‐10 by VEGF in HUVEC and supports an angiogenic role for MMP‐10 in response to VEGF stimulation in vitro and in vivo.
Why not high‐throughput eukaryotic protein structures?
Authors: Fox Brian Grant , Volkman Brian F , Phillips George N , Markley John L
Published: June 2021

During the Phase I and Phase II of the Protein Structure Initiative, the Center for Eukaryotic Structural Genomics focused on developing approaches for high‐throughput structure determination by working with eukaryotics, including humans. Around 9,000 genes were examined, over 1,000 proteins purified to date, and 127 structures determined. Outside requests for studies of over 500 proteins were evaluated. This work suggests expanded efforts with eukaryotic proteins in PSI:Biology will be feasible and can yield many new insights that link structure and function. Based on our experience, effective attack on challenges represented by eukaryotic proteins requires many approaches at all steps in the pipeline from bioinformatics to structure determination. Examples of how different experimental approaches developed by this project are used to prepare and characterize eukaryotic proteins for structure determination will be presented, along with the challenges and opportunities encountered along this path. This work was supported by NIH Protein Structure Initiative Grants P50 GM064598 and NIH U54‐GM074901 (J.L. Markley, PI). The authors thank all members of the CESG team.
Non‐nutritional modulation of cobalamin levels: TCN1 mutations, polymorphisms, and alternative splicing, and FUT2 polymorphism
Authors: Carmel Ralph , Parker James , Kelman Zvi
Published: June 2021

A nutritional explanation is usually affixed to cobalamin (B12) level variations, but we have linked many low B12 levels to deficiency of transcobalamin (TC) I, which carries >80% of plasma B12, and detected 4 relevant mutations of the TCN1 gene. We now present a more extended network of genetic influences on B12 via TC I.

Methods

TCN1 mutations and mRNA splicing variants were sequenced in 55 subjects (8 had exon 2 nonsense mutations 270delG or 315C>T, and 10 had a 999G>T polymorphism in exon 6). We also tested a FUT2 polymorphism whose homozygous state is associated with lower 1,2‐fucosyltransferase activity and higher serum B12.

Results

Most TCN1 splice variants began with exon 2 deletion, often extending to exon 3 or beyond without a frame shift, or with exon 5 deletion, usually extending through exon 7. The 18 TCN1 mutations were all associated with low TC I and B12 levels, excepting only the sole person who was also homozygous for FUT2 polymorphism (his relatives had identical results).

Conclusions

1. TCN1 splice variations usually start in exon 2 or 5 and can extend downstream with varying consequences. 2. If exon 2 contains nonsense mutations, its splicing out may prevent nonsense‐mediated decay of TC I. 3. FUT2 may affect fucosylation of TC I, a heavily glycosylated protein. 4. FUT2 polymorphism modulates the low TC I and B12 levels caused by TCN1 mutations.
Optimizing growth of Lactobacillus reuteri in phytone peptone‐tryptone‐glucose medium by using response surface methodology
Authors: Atilola Olabiyi Akinbami , Ibrahim Salam , Song Danfeng
Published: June 2021

Lactobacillus reuteri as probiotic helps prevent and treat both viral and bacterial diarrhea enhancing the body's resistance to gastrointestinal disease. However, the growth of L. reuteri is very limited and certain nutrients are needed for optimum growth and high cell density. The objectives of this study were to determine the effect of different protein sources on growth and bacterial cell density of L. reuteri and to assess the optimum growth requirements using response surface methodology. Four strains (CF 2‐7F, MF 14 C, DSM 20016, and SD 2112) of L. reuteri were used in this study. Overnight cultures were inoculated into fresh basal growth broth with various proteins sources (peptone, tryptone, proteose peptone, phytone peptone, tryptic soy broth, yeast extract, and beef extract). Samples were then mixed well and incubated at 37 °C for 48 hrs. Bacterial growth was monitored by measuring turbidity at 610 nm in a spectrophotometer at different time intervals during the incubation period. At the end of incubation, samples were plated on MRS agar to determine the bacterial population. Our results showed that phytone peptone or tryptone promoted the growth of L. reuteri (P < 0. 01) compared to the control. The response surface model indicated that phytone peptone (5–10 g/L), tryptone (5 g/L), and glucose (15g/L) could be used as optimum concentrations to enhance the growth and cell density of L. reuteri in growth medium.
Is leucine induced p70S6 kinase phosphorylation following resistance exercise dependent on elevated phenylalanine levels in human skeletal muscle?
Authors: Apró William , Holmberg Hans‐Christer , Rooyackers Olav , Blomstrand Eva
Published: June 2021

The purpose of this study was to investigate the specific role of leucine in the stimulation of the mammalian target of rapamycin signalling pathway. Six male subjects performed four heavy resistance exercise sessions, each separated by approximately one week. Subjects were randomly supplemented with one of four drinks: placebo (flavored water), leucine or essential amino acids (EAA) with and without leucine. Immediately following each exercise session, four subjects were infused with a flooding dose of L‐[2H5] phenylalanine (Inf) while two subjects served as controls (Ctrl). Muscle biopsies were taken before and one hour after exercise. In the Ctrl group, resistance exercise resulted in a substantial increase (45‐fold) in p70S6 kinase phosphorylation when all EAA were ingested, whereas ingestion of leucine alone had no greater effect than that of placebo. In the Inf group, however, ingestion of leucine alone and EAA increased p70S6k phosphorylation to a similar extent (35‐fold). The divergent signalling response in the two groups suggests that leucine alone is insufficient to increase p70S6k phosphorylation. Indeed, in the Inf group, there was a strong correlation (r=0.91) between p70S6kphosphorylation and the product of muscle leucine and phenylalanine levels. These results suggest that the stimulatory effect of leucine on p70S6kphosphorylation is dependent on elevated muscle phenylalanine levels. Supported by the Swedish National Centre for Research in Sports
Total (TAA) and Free Amino Acid (FAA) Profile in Term and Preterm Human Milk (HM) through the Course of Lactation: a Systematic Review and Meta‐Analysis
Authors: Zhang Zhi‐Ying , Adelman Alicia Suzanne , Rai Deshanie
Published: June 2021

Amino acid profile and total nitrogen are key aspects of HM protein quality. Herein, we report a systematic review and meta‐analysis of published FAA and TAA profiles in HM from several countries. Analysis of Variance and Principle Components Analysis were used to analyze the effects of gestation period, lactation stage, and geographical region. Data on total nitrogen and TAA composition of HM revealed general inter‐study consistency whereas FAA concentrations varied substantially among studies. All 18 individual TAA concentrations in HM steadily declined in the first two months of lactation and remained relatively unchanged thereafter through the study period of 18 months. In contrast, the levels of the FAA glutamic acid and glutamine peaked at 3–6 months and decreased thereafter. Interestingly, glutamic acid was present at ~50‐fold higher level compared to other FAA in HM. Statistically significant differences were observed for some of the TAA and FAA between preterm and term milk as well as among different regions, i.e. North America vs. Asia Pacific vs. Europe; there was a paucity of data from Africa. This systematic review represents a useful data set for the evaluation of protein quality of breast milk substitutes for preterm and term infants. Its comprehensive nature may serve as a guide to support future efforts in further development of global and/or regional human milk amino acid standards.
Meal‐replacement shake and intermittent fasting combined with herbal beverage leads to healthy weight loss
Authors: Rockway Susie W. , Balliett Mary , Burke Jean , Rasmussen Oscar
Published: June 2021

Aim

This pilot study evaluates the impact of a novel whey‐based meal replacement shake combined with herbal beverage instead of pure fasting on weight in healthy adults.

Design

In a 28‐day study, a total of 30 subjects, ages 20 to 60, meeting inclusion criteria replaced 2 of 3 meals per day with a whey‐based beverage, consumed a 400–600 kcal meal per day with instruction to avoid foods with high‐glycemic load, and rather than a pure water‐fast 1 day per week, subjects consumed 16 oz of an herbal beverage along with water. Baseline and post‐treatment anthropometric measurements and blood were taken. Subjects completed 100mm visual analog scale (VAS) measuring perception of weight, gastrointestinal upset, and energy level at baseline and day 28; dietary intake and adverse effects were recorded daily in a food diary.

Statistics

Dependent, quantitative variables were tested for significance using paired t‐tests.

Results

After 28 days, subjects lost an average of 6.4 lbs (3.6% of initial body weight), which was statistically significant (p<0.05) from baseline weight (175.9 ± 36.8). Waist and hip circumference decreased by 1.9 and 1.1 inches, respectively and were statistically different from baseline. Total cholesterol and LDL cholesterol were significantly decreased (13.30mg/dL and 11.40 mg/dl), respectively.

Products and funding for this study was provided by Isagenix, International
Early childhood multiple micronutrient supplementation is associated with lower obesity prevalence in later childhood, compared with iron and vitamin A supplementation only
Authors: Harding Kimberly B , Ramakrishnan Usha , Mejia Fabiola , Guerra Armando Garcia , Martorell Reynaldo , Neufeld Lynnette M
Published: June 2021

We examined whether multiple micronutrient (MM) supplementation in early childhood has any lasting effect on child size in later childhood. In a randomized, double‐blind controlled trial in Mexico, infants (n=650) from a prenatal supplementation trial (MM or iron only) received either MM or iron and vitamin A (Fe‐A) supplements for 6d/wk from 3–24mo of age. Child height (ht) and weight (wt) were measured in a 2008 follow‐up visit (n=607; mean age 8.5 ± 0.9y). Using intent to treat analysis [4 group comparisons (by mother and child group) and by child group only] we found no significant differences (p>.1) in child wt, ht, ht‐for‐age or wt‐for‐age Z‐scores (HAZ or WAZ), or prevalence of stunting (HAZ<−2), underweight (WAZ<−2), thinness (BMI‐forage Z‐score (BAZ)< −2) or overweight (BAZ +1 to +1.9). However, the child MM group had significantly lower BMI, BAZ and obesity prevalence (BAZ>+2) than the child Fe‐A group (mean BMI 17.3 ± 2.7 vs. 17.8 ± 2.9 kg/m2, mean BAZ 0.5 ± 1.1 vs. 0.8 ± 1.2, obesity 10.2 vs. 17.3%; all p<.05). Analysis by 4 groups showed a similar, though not statistically significant trend (p=.07); children who received MM (in utero and early childhood) had the lowest obesity prevalence (9.2%) and those who received Fe only in utero and Fe‐A in childhood had the highest prevalence (18.3%). Results are consistent with our findings at 2y of age and suggest long term benefits of MM supplementation in early life. Further analysis is required to adjust for contextual factors that may have contributed to this difference after supplementation was completed.

Funding: MI; Thrasher Research Fund; UNICEF; CONACYT; INSP Mexico; UC Berkeley; Emory University
Anatomy Virtual Tutor: an ipod touch application for the first year medical student
Authors: Rahimi Omid B , Rahimi Navid
Published: June 2021

For many freshmen medical students, the gross anatomy laboratory exam is their first experience in taking a practical type examination. The goal of this project is to develop an interactive medium to help students appreciate how anatomical structures are displayed during examination by focusing student's attention on important anatomical relationships. With the prevalence of digital media player use among students and more than 100K applications created by developers for the iphone and ipod touch, Apple's ipod pocket computer provides a unique and ubiquitous platform to distribute digital educational content. We have developed an ipod application which allows the student to study gross anatomical structures on still photographs of cadaveric dissections. The student may choose a structure from a hit list or enter “quiz” mode, which asks the student to identify a structure. The application is divided into five content categories that span Head & Neck, Thorax, Abdomen, Pelvis, and Extremities. The student may select a category to review images and structures for the particular anatomical region. This application allows students to assess their level of competence by taking a virtual practical via the quiz mode and enhances their anatomical knowledge by emphasizing relationships among structures visualized in a dissection field.
Appropriate waist circumference cutoff values for the diagnosis of metabolic syndrome in Mexican American adults
Authors: Sarafrazi Neda , Jackson Robert T , Momen Bahram
Published: June 2021

Metabolic syndrome (MetS) increases the risk of cardiovascular disease and diabetes. The International Diabetes Federation (IDF) recently proposed new criteria for the diagnosis of MetS, which requires the presence of central obesity as measured by ethnic specific waist circumference (WC) cutoff values. Currently, no specific WC thresholds for diagnosis of central obesity in Hispanics are available. The objectives were to determine the appropriate gender specific WC thresholds for diagnosis of central obesity in Mexican American (MA) adults and to estimate the prevalence of MetS using IDF definition with and without the modified WC in this population. Data from 3265 MA adults aged 20–80 years who participated in the National Health and Nutrition Examination Survey 1999–2006 were used. The prevalence of MetS was compared using IDF criteria with and without the modified waist circumference. Receiver operating characteristic curve analysis suggested that yielding at least 80% sensitivity, the WC value of 90 cm in both genders was more appropriate in predicting the presence of two or more MetS risk factors in this population. Based on our cutoff, there was 34% reduction in the prevalence of central obesity in women (82.5% to 54.2%). The age adjusted prevalence of MetS decreased from 58.4 to 48.2%. The MetS was more common among MA men than women (55.8% in men versus 37.8% in women, P =0.0003). Our findings provided a practical guidance in the assessment and screening of central obesity MetS in Mexican Americans.
Design and implementation of the new UK‐WHO growth charts
Authors: Wright Charlotte Margaret , Haines Linda , Reddy Sheela
Published: June 2021

Growth charts are essential for nutritional assessment in childhood. The UK were early adopters of the new WHO standards and used this opportunity to completely redesign the layout and instructions of charts for children up to age 4 years.

Method

The design team drew on existing literature, new data analyses and product testing with parents and health staff focus groups, using case scenarios to assess plotting competency and understanding. Instructions were then written and tested with subsequent focus groups.

Results

The charts present preterm (using UK 1990 reference) and infant (WHO) charts separately, up to age 4. The 50th centile is de‐emphasized to encourage parents to understand the wide range of normal growth. Other innovations include a BMI look up and adult height predictor, which avoid the need for calculation. The focus groups revealed high levels of plotting errors, inconsistent practice, and high demand for formal training. These findings have informed a set of standardized supporting training and educational materials, free to download from www.growthcharts.rcpch.ac.uk. The charts were published in May 2009 and are now being issued to new births in England and Scotland.

Conclusions

The UK‐WHO growth charts are the first national charts to have been developed using an evidenced based approach worthy of the unique WHO data set.

Funding: Department of Health
Maternal zinc intake during pregnancy is positively associated with birth weight
Authors: Lee Yo A , Kim Hyesook , Ha Eun‐hee , Park Hyesook , Ha Mina , Kim Yangho , Hong Yun‐Chul , Chang Namsoo
Published: June 2021

There have been studies about the relationship between zinc intake and pregnancy outcomes, but the results are inconsistent. We examined the association between maternal zinc intake and pregnancy outcomes in pregnant women enrolled in the Mothers and Children's Environmental Health cohort study. Dietary intake of midterm pregnancy was assessed by a 24‐hour recall method and pregnancy outcome data were collected from medical records. Total zinc intake was 8.4mg/day and that from animal‐sources was 3.4mg/day. A significantly difference in birth weight and height was associated with intakes of increasing amounts of zinc from animal‐source (P for trend <0.01 for birth weight and P for trend <0.001 for birth height, respectively) but not zinc from plant‐sources after adjustment for the covariates. Multiple regression analysis with covariates revealed that birth weight was associated with total zinc intake (P=0.001) and zinc intake from animal‐source (P=0.035), respectively. In conclusion, we found that group for animal‐source with good bioavailability is positively associated with birth weight and height.

This work was supported by the Ministry of Environment and the Brain Korea 21 project in 2009.
Food experience in infancy and unhealthy eating behavior in later childhood
Authors: Shim Jae Eun , Kim Juhee
Published: June 2021

This study was performed to evaluate infant feeding practices potentially related to unhealthy eating behavior (UEB) in later life. The STRONG Kids project conducted a parent survey for children aged 2–3 years old enrolled in child care center in Champaign County, IL. A total of 242 parents from the wave 1 study completed a self‐administered survey and 218 valid ones were analyzed. We tested breastfeeding, formula feeding, and introduction of complementary foods during infancy and the relation with parent's perception of child's current UEB. Approximately 51% of children didn't experience home‐made baby food (HBF); 31% didn't have table food (TF); 17% didn't have both. We considered that no experience with HBF and TF in infancy to be an indicator of failure of exposure to various texture and flavor in that period and defined it as an inappropriate food experience (IFE). About 34% of children with IFE were perceived as picky eaters and 25% in the non‐IFE; 37% of those with IFE were perceived to barely accept new food and 21% in the non‐IFE (p=0.0312); 18% of those with IFE were perceived to eat limited variety of foods and 20% in the non‐IFE; 8% of those with IFE were perceived to prefer specific food preparation and 7% in the non‐IFE. Our study indicated that various food experiences in infancy could promote child's acceptance of new food in later life.

Funded by Illinois C‐FAR grant
Mouse Muscle‐Derived Stem Cells in a Murine Model of Accelerated Aging
Authors: Lavasani Mitra , Robinson Andria R , Lu Aiping , Feduska Joseph M , Tilstra Jeremy S , Drowley Lauren , Robbins Paul D , Niedernhofer Laura J , Huard Johnny
Published: June 2021

Aging is characterized by the progressive erosion of tissue homeostasis and functional reserve in all organ systems. Although controversy remains as to the molecular mechanism(s) underlying the process of aging, accumulated cellular damage, including DNA damage, appears to be a major determinant of lifespan as well as age‐related pathologies. Moreover, there is evidence that accumulation of damage in stem cells renders them defective of self‐renewing and regenerating damaged tissues. Here we test this in a murine model of a progeroid syndrome, a disease of accelerated aging. Post‐natal muscle‐derived stem cells (MDSCs) from the progeroid mice were defective in proliferation and myogenic differentiation, and hypersensitive to oxidative stress. Intraperitoneal delivery of wild‐type (WT) MDSCs resulted in a significant lifespan and healthspan extension in progeroid mouse models. Our results also demonstrate significant improvement in vascularization of the muscle as well as muscle fiber size. Interestingly, the proliferation and myogenic differentiation of progeroid ERCC1‐deficient MDSCs were significantly improved when co‐cultured with WT‐MDSCs. These results provide evidence for aging‐related loss of functional stem cells and demonstrate the therapeutic potential of stem cells to delay onset of age‐related degenerative diseases. This work was supported by The NIEHS and NIH.
Hair Regeneration Effect of Korean Panax Ginseng Berry in C57BL/6 mice
Authors: Park Soojin , Ryu Jin‐A , Jeong Min‐Ah
Published: June 2021

Korean ginseng (Panax ginseng C. A. Meyer) root have been documented to possess hair growth effect in humans. However, studies on the Korean ginseng berry (KGB) are sparse. To evaluate the effect of KGB on hair regeneration in C57BL/6, animals were divided into 4 groups including vehicle control (30% ethanol), positive control (5% minoxidil) and experimental groups (KGB, 30 and 60 mg/ml). And they were topically applied respective treatments on the depilated dorsal skin with an amount of 100 ul everyday for 4 weeks. Hair regeneration was evaluated by photographical and histological observations. KGB groups promoted development of hair follicles approximately 20% more than vehicle group through the stimulation of anagen progression of the hair shaft. Interestingly, hair reconstitution was more effective in low dose KGB group than high dose KGB group. The major ginsenoside was Re (5.99%) in total ginsenoside (9.09%) in KGB, analyzed by HPLC, which were 28 times more than that of Korean ginseng root. In conclusion, our results implicate the enriching potential of KGB for hair regeneration. A further study will present whether the synergistic effect of dietary and topical administrations of KGB on hair restoration in 3‐year project. This work was supported by the National Research Foundation of Korea (NRF) grant funded by the Korea government (MEST) (No. 2009‐0067117).
Evaluation of 12‐week body weight reduction program in obese Korean adults
Authors: Kim Junglim , Jee Jaehwan , Choi Yoonho , Lee Seonheui
Published: June 2021

The purpose of this study was to evaluate the effect of a moderate weight reduction program provided by a medical center in Seoul in 79 obese Korean subjects. The program was 12 weeks long and composed of low calorie diet, exercise, and behavior modification. The participants visited the medical center once every 2 weeks for 12 weeks. Dietary habits, intakes and body composition were assessed before and after the program. On the completion of 12‐week program, dietary intakes of energy, protein and fat were significantly decreased (p<0.01), but the index of nutritional quality (INQ) was increased. After 12 weeks, the percentage of participants in the normal weight category had increased from 7.6% to 20.2%, while average weight decreased from 81.2±12.8 kg to 75.3±11.6 kg. The dietary habits such as regularity of mealtime and overeating were improved. Daily intakes of fatty meat, sweet snack and coffee decreased, but the consumption of tofu, vegetables and green tea increased (p<0.05). Follow‐up program participants(n=55) when they visited for health check‐up, maintained 99.9% of their reduced weight(mean 19.8 months). This study implies that a consistent weight reduction program is effective to reduce body weight and body fat. Further the results indicate that improvement of dietary behavior such as dietary habits, food consumption pattern could help maintain the reduced weight successfully. No funding relating to this abstract.