bluuebunny/crossref_metadata_2025_split huggingface dataset:
Nitric oxide synthase inhibition prevents activity‐induced NFAT nuclear accumulation and skeletal muscle adaptation
Authors: Martins Karen JB , St‐Louis Mathieu , Michel Robin N , Murdoch Gordon K , Dixon Walter T , MacLean Ian M , McDonald Pamela , Putman Charles T 
Published: June 2021

Involvement of calcineurin/NFAT signaling in the regulation of skeletal muscle myosin heavy chain (MHC)‐based fast‐to‐slower isoform adaptations (F‐S) is recognized. Less is known about the role nitric oxide (NO) plays in this process.OBJECTIVETo investigate the necessity of NO in activity‐induced skeletal muscle F‐S in vivo.METHODSEndogenous NO production was blocked by administering L‐NAME (0.75 mg ml−1; ~100 mg kg−1 day−1) during 0, 1, 2, 5 or 10 days of chronic low‐frequency stimulation (CLFS; 10 Hz, 12 h d−1) applied to the tibialis anterior and extensor digitorum longus muscles of rats (L‐Stim; n=6 each group). Control rats received only CLFS (Stim; n=6 each group).RESULTSCLFS induced increases in NFATc1 nuclear localization in Stim, which did not occur in L‐Stim at any time point. These results were confirmed by western blot analyses. Moreover, MHC mRNA, protein and fibre type analyses revealed CLFS‐induced F‐S occurred in Stim, but were completely abolished in L‐Stim.CONCLUSIONSNO may be regulating activity‐induced MHC‐based F‐S at the transcriptional level via NFAT nuclear accumulation in vivo. *Authors contributed equally. Funded by NSERC, AHFMR, CIHR and CRC.
Effects of ganoderma lucidum spores on expression of PSA‐NCAM and hippocampus mossy fiber sprouting in pentylenetetrazol‐induced epileptic rats
Authors: Li Xiao‐Jie , Wang Shu‐Qiu , Zhang Shi‐Ling , Kang Yu‐Ming 
Published: June 2021

The present study was undertaken to investigate the effects of ganoderma lucidum spores (GLS) on expression of polysialic acid neural cell adhesion molecule (PSA‐NCAM) and hippocampus mossy fiber sprouting (MFS) in pentylenetetrazol (PTZ)‐induced epileptic rat model. Thirty male Sprague‐Dawley rats (180–220g) were divided into three groups: control, GLS and epilepsy model groups. The results showed that, compared with epilepsy model groups, the rats of GLS group have longer latent periods, and lower seizure level. Their seizure durations also were shorter than epilepsy model group. In the granular cell layer of dentate gyrus and stratum pyramidal and stratum oriens of the CA3 region, epilepsy model group have more PSA‐NCAM than GLS group and control group; compared with epilepsy model group, MFS in the granular cell layer of dentate gyrus were obviously less in GLS group, and no statistically significant difference was detected when compared with control group; in the stratum pyramidal and stratum oriens of the CA3 region, MFS in GLS group was less than epilepsy model group, but obviously more than control group.ConclusionFor PTZ induced epileptic rats, there were different severities of MFS in different regions of hippocampus. GLS can decrease expression of PSA‐NCAM and MFS effectively, and that might be one of its mechanism of lighten rats′ seizure induced by PTZ.
Tricellulin in Crohn's disease and ulcerative colitis
Authors: Krug Susanne M , Bojarski Christian , Fromm Anja , Schulzke Jörg Dieter , Fromm Michael 
Published: June 2021

The tight junction (TJ) protein tricellulina (TRIC) is predominantly located in tricellular TJs (tTJ), the meeting points of three cells. There it forms a barrier to macromolecules without affecting ion permeability. However, at low TRIC levels the tTJ forms a pathway for macromolecules. We studied the pathologic relevance of TRIC expression in Crohn's disease (CD) and ulcerative colitis (UC).MethodsDilution potentials and permeabilities for fluorescein and FITC‐dextran 4K were measured with the Ussing technique in sigmoid colon biopsies from controls, active CD, and UC patients. Epithelial and subepithelial resistances were determined by impedance spectroscopy. Expression of TRIC and other TJ proteins were analyzed by Western blot.ResultsExpression of TRIC was reduced in sigmoid colon of UC patients, but not in CD samples. This was in parallel to known changes in expression of e.g. claudin‐4. UC specimens exhibited decreased epithelial resistance, increased subepithelial resistance, and unchanged transepithelial resistance and Na+ and Cl− permeability. In contrast, permeabilities for fluorescein and FITC‐dextran 4K were increased.ConclusionTRIC is downregulated in UC, while it is not influenced in CD. As TRIC limits the macromolecular passage in tTJs, its downregulation may contribute to the increased macromolecular permeability in UC. (Supported by DFG FOR 721)
Expression of the Crohn's disease candidate gene, PTPN2, is upregulated in active Crohn's disease
Authors: Scharl Michael , Weber Achim , Hausmann Martin , Rogler Gerhard , McCole Declan F 
Published: June 2021

We have recently shown that IFNγ, a major cytokine involved in Crohn's disease (CD), increases expression and activity of the CD candidate gene, PTPN2, in intestinal epithelial cells (IEC). We have also shown that PTPN2 restricts IFNγ‐induced IEC permeability. Here, we investigated whether PTPN2 expression is altered in active CD. mRNA analysis was performed by real‐time PCR. Immunostaining was performed on formalin‐fixed mucosal biopsies. We investigated PTPN2 mRNA levels in biopsies from macroscopically non‐inflamed areas of the terminal ileum, colon or rectum of CD patients in clinical and macroscopic remission; macroscopically inflamed areas of the terminal ileum and colon of CD patients with clinically active disease; or healthy control subjects undergoing routine screening. We observed significantly elevated PTPN2 mRNA in active CD (p<0.05; n=9), but not in CD patients in remission. Immunostaining showed elevated epithelial and lamina propria PTPN2 in CD vs. controls. CD patients with active disease, but not those in remission, also showed an approximately three‐fold increase in tissue IFNγ mRNA levels compared to healthy controls. H&E‐stained biopsy specimens indicated acute inflammation in patients with active CD only. Overall, our findings demonstrate altered PTPN2 expression in active CD and correlate with observed effects of IFNγ on PTPN2 expression in vitro.Supported by the CCFA and DFG.
Quantification of serum fucosylated glycoproteins as lung adenocarcinoma biomarkers using AlphaLISA technology
Authors: Chow Lu‐Ping , Wen Chu‐Ling 
Published: June 2021

Lung cancer is the leading cause of cancer‐related death in Taiwan for recent years. At the time of diagnosis, most of the lung cancer patients are at advanced stages; therefore, new diagnostic strategies detecting the early stage cancer have been eagerly expected. There is increasing evidence that glycosylation plays a significant role in biological processes including oncogenic transformation and metastasis. Several reports suggested that the alterations in protein glycosylation might be associated with the development of certain types of human cancer.The specific aim of our study is to setup a platform to screen and identify serum glycoproteins as biomarkers of lung cancer. To enrich N‐linked glycoproteins from serum samples, Con A affinity chromatography were established. The captured glycoproteins were tryptic digested and then analyzed by LC‐MS/MS. For high through‐put screening, we established an AlphaLISA assay, which is a bead‐based proximity assay, to quantify the fucosylation level of these glycoproteins.Comparison of the serum glycoproteom of normal controls and adenocarcinoma patients revealed some cancer‐related proteins. In addition, the changes of composition and amount of the N‐linked oligosaccharide were evaluated in the sera of patients by lectin staining and AlphaLISA.
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
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 ObjectiveA 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.MethodsHuman 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.ResultsIn 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.
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.
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.
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.MethodsTCN1 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.ResultsMost 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).Conclusions1. 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.
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

AimThis 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.DesignIn 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.StatisticsDependent, quantitative variables were tested for significance using paired t‐tests.ResultsAfter 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
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.MethodThe 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.ResultsThe 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.ConclusionsThe 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