bluuebunny/crossref_metadata_2025_split huggingface dataset:
Linagliptin, a dipeptidylpeptidase 4 inhibitor, ameliorates indomethacin‐induced enteropathy via GLP‐2 pathway
Authors: Akiba Yasutada , Kaunitz Jonathan 
Published: June 2021

Glucagon‐like peptide‐2 (GLP‐2) is an intestinotrophic hormone secreted from enteroendocrine L cells. We have reported that dipeptidylpeptidase 4 (DPP4) inhibition prevents formation and promotes healing of indomethacin (IND)‐induced small intestinal ulcers in rats via GLP‐2 release and GLP‐2 receptor activation. We thus further examined the effect of the clinical DPP4 inhibitor linagliptin (LNG) on IND‐induced enteropathy.Small intestinal ulcers were induced by IND treatment (10 mg/kg, sc) in rats. LNG (0.1 – 5 mg/kg) was given once before IND for prevention, and for healing, once daily for 2 days starting 1 day after IND treatment.LNG treatment dose‐dependently reduced ulcer scores, accompanied by an increase of portal venous GLP‐2 concentrations. LNG promoted ulcer healing of IND‐induced intestinal ulcers. LNG‐induced healing was reversed by a GLP‐2 receptor antagonist. Ulcer healing was further accelerated by co‐treatment with L‐alanine plus inosinate, an umami taste receptor agonist. Compared to LNG, the DPP4 inhibitor NVP‐728, metabolized in the kidney, had a little effect on the enteropathy, suggesting that a long‐acting, entero‐hepatically circulated DPP4 inhibitor may be more efficacious in the prevention of intestinal mucosal damage.These results confirm our previous study that DPP4 inhibition prevents ulceration and promotes ulcer healing of NSAID‐induced enteropathy, enhanced by the luminal umami taste receptor activation, suggesting that GLP‐2 release enhanced by DPP4 inhibition and nutrient receptor activation is useful for the treatment of NSAID‐induced enteropathy.Supported by Boehringer Ingelheim, VA Merit Review, NIH R01 DK54221
The Role of Suppressor of Cytokine Signalling 3 (SOCS3) in the Regulation of Intestinal Epithelial Cell Turnover in a Helminth Model
Authors: Shaw Elisabeth , Smith Emily , Else Kathryn , Rigby Rachael 
Published: June 2021

Gastrointestinal epithelial cells (IEC) act as a first line of defence against invading pathogens and are replenished every 4‐5 days. An increase in IEC turnover aids the expulsion of intestinal pathogens including helminth infection. SOCS3 limits IEC proliferation with its loss associated with hyperproliferation and increased tumour growth in intestinal injury and cancer models. This study investigates SOCS3 effects on IEC turnover, outcome of infection and biochemical changes in response to infection in an in vivo Trichuris muris model.Mice with IEC SOCS3 conditional knockdown and controls were infected with T. muris. Crypt depth, cell proliferation rates, worm burden and immune response were assessed. Protein expression assessed by western blotting or immunohistochemistry.Increased SOCS3 expression was observed in cecum of T. muris susceptible vs. resistant mice strains. This supported our hypothesis that increased SOCS3 may limit IEC turnover and parasite expulsion. IEC deletion of SOCS3 did not lead to resistance to infection although a trend towards increased expulsion, compared to controls, was observed. At 35 days post infection an increase in proliferation of cells in cecal crypts was observed with no change in crypt depth in knockdown animals, supporting an increase in cell turnover. No difference in crypt dynamics was apparent in early stages of T. muris infection. Loss of SOCS3 was associated with altered indoleamine 2,3‐dioxygenase (IDO) expression in goblet cells.During parasitic challenge SOCS3 modulates IEC proliferation, potentially via a mechanism mediated though altered IDO.Medical Research Council G1100211/1
Development of a Highly Specific Met Antibody for Screening Met Overexpression in NSCLC
Authors: Chen Caiwei , Wei Haitao , Yuan Kehu , Ma Donghui , He Wei‐wu 
Published: June 2021

Approximately 5%‐22% of non‐small cell lung cancer (NSCLC) patients with secondary resistance to EGFR TKIs had evidence of amplification of the Met oncogene. This dysregulation of Met signaling was implicated in the proliferation and eventual metastasis of NSCLC, and therefore represents an attractive target for anticancer drug development.Currently, there are a number of clinical trials for Met targeted small molecule and monoclonal antibody therapies. Immunohistochemistry (IHC) analysis of the Met protein will be very important for these Met targeted therapies and requires a highly specific antibody to prevent false positives. To develop a highly specific monoclonal antibody against Met for IHC analysis in NSCLC, we generated 97 monoclonal antibodies. IHC evaluation showed that 8 of the antibodies had suitable performance in Met positive NSCLC patient tissues. The 8 Met monoclonal antibodies were further tested on a high density protein microarray chip containing 17,000 human proteins. The microarray analysis showed that clone 3G5 was highly specific as evidenced with it only reacting with Met on the microarray. This new 3G5 Met monoclonal antibody will be a promising candidate as a diagnostic reagent for detecting Met protein in NSCLC with minimal false positives.
Isolation of Human Glioblastoma Multiforme Primary Cells and analysis of CD133+
Authors: Yukselten Yunus , Akcora Yildiz Dilara , Demircan Kadir , Sunguroglu Asuman , Ugur Hasan 
Published: June 2021

ObjectiveGlioblastoma multiforme (GBM) is an incurable and the most common type of lethal brain tumors. A transmembrane glycoprotein CD133 has been used as a marker in cancer stem cell analysis.In this study, we aimed isolation of GBM Primary Cells from human GBM tumor tissue which is surgically resected and to determine amount of CD133+ in the primary cultures with using AC133 monoclonal antibody.Material Method : Primer Cell cultureTumor tissues taken from three patients who had GBM diagnosis were separated into 1 mm3 fractions with bistoury within fresh medium. 1ml of DMEM mixture were seeded into T‐25 cm2 culture flasks and incubated at 37°C with 5% CO2.CD133+ stainingCD133+ staining was used as a marker of GBM stem cell marker. Cells were method according to cell surface staining.ResultsWe demonstrated the GBM primary culture isolation of tumor tissues taken from four patients who had GBM diagnosis. We studied three GBM primary cultures such as P1, P2, and P3. We performed amount of CD133+ cells in GBM primary cell cultures by using the method of FCM analysis. According to the FCM analysis results in %1,5 ‐% 1,6 ‐%1,3 are founded respectively.ConclusionFinally, Isolated GBM primary cells provide an invaluable mediate to study how tumors form, progress, and relapse and also to elucidate the underlying deriving cellular mechanisms that eventually could provide understandings to therapeutic options. GBM Stem Cells (GSC) are responsible for tumor generation and treatment resistance in GBM. Identifying GSC cells is an important step in GBM therapy.
Production of Fluorescent Oligosaccharide Substrates for the Neisseria meningitidis Serogroup W Capsule Polymerase
Authors: McCarthy Pumtiwitt , Williams Corshai , Ukaegbu Ophelia , Gordon‐Travers Tresha 
Published: June 2021

Neisseria meningitidis is a Gram‐negative bacteria that causes most cases of bacterial meningitis. The six disease‐causing serogroups of N. meningitidis differ in capsular polysaccharide composition and linkage. Our long‐term goal is to develop a better understanding of the molecular mechanism of the serogroup W capsule polymerase. This enzyme synthesizes a heteropolysaccharide of galactose and sialic acid. We aim to develop a fluorescence‐based assay to aid in our studies. In this work, we describe our initial efforts to produce fluorescent substrates for the serogroup W capsule polymerase. Preliminary radioactivity assays indicate the enzyme has the ability to elongate serogroup W oligosaccharide fragments. The enzyme was more active with oligosaccharides than full length polysaccharide. These fragments, obtained by acid‐hydrolysis and heating, are the starting point for production of fluorescent substrates. Oligosaccharides were subjected to reaction with the fluorescent dye 1,2‐diamino‐4,5‐methylenedioxybenzene (DMB). Initial results indicate successful incorporation of the dye as assessed by HPLC with fluorescence detection. Alternately, copper‐catalyzed “click” chemistry is also being used to produce fluorescent oligosaccharides. Chemical reactions are being performed to add an azido group to oligosaccharides. These will be conjugated to an alkyne‐containing dye by 1,3‐cycloaddition “click” chemistry. Future work seeks to further characterize and optimize fluorescent substrates from both methods for testing with the serogroup W capsule polymerase.Funded in part by ACS Project SEED (C.W).
Enzymogenesis of Zinc‐Dependent Alcohol Dehydrogenases and Aldehyde Dehydrogenases in Animals Show That Their Origin Predates Natural Availability of Ethanol from the Diet
Authors: Julián‐Sánchez Adriana , Hernández‐Tobías Aída , Vara‐Cisneros Paola , Gaona‐López Carlos , Alvarado‐Leaño Juan , Piña Enrique , Riveros‐Rosas Hector 
Published: June 2021

In vertebrates, ethanol metabolism is performed through the concerted action of two enzymes: a Zn‐dependent alcohol dehydrogenase (Zn‐ADH) that oxidizes ethanol to acetaldehyde, and an aldehyde dehydrogenase (ALDH) that oxidizes acetaldehyde to acetate. The presence of these Zn‐ADHs and several ALDHs in animals has been assumed to be a consequence of chronic ethanol exposure from the diet. The main natural source of ethanol is fermentation of fruit sugars by yeast. The origin of angiosperms with fleshy fruits dates from the Lower Cretaceous (130‐140 million years ago). Because association between yeast and angiosperms dates also from the Cretaceous, dietary exposure of diverse frugivorous taxa to ethanol is similarly ancient. Since substrate availability must precede enzyme selection, then it can be expected that the enzymogenesis of the ADHs/ALDHs began after the appearance of fleshy fruits. Phylogenetic analyses reveal that all different Zn‐ADHs (seven classes) in animals appeared before land plants, independently of ethanol availability. With respect to aldehyde dehydrogenases (ALDH), phylogenetic analyses showed 13 different ALDH families in animals, and all they appeared before land plants. Because all these enzymes are not induced by ethanol and possess a high activity with non‐ethanol endogenous substrates, it can be concluded that Zn‐ADH and ALDH participation in ethanol metabolism can be considered as incidental, and not adaptive.Supported by DGAPA‐UNAM grant IN216513.
Reconstructing Membrane‐Anchored Mitochondrial AAA+ Proteases For In Vitro Analysis
Authors: Glynn Steven , Rampello Anthony , Shi Hui 
Published: June 2021

Mitochondrial AAA proteases harness energy from ATP hydrolysis to remove and degrade damaged protein components of the mitochondrial inner membrane. Transmembrane domains anchor the proteases in the inner membrane and drive assembly of the active hexameric form of the enzymes. To date, the requirement for these transmembrane domains has hampered detailed analysis of the substrate binding and processing activities of these important enzymes. The objective of this study is to develop a system for reconstituting membrane‐anchored ATP‐dependent proteases in a soluble, assembled, and active form that can be used to elucidate the molecular mechanisms of these important enzymes.We report a novel method for reconstructing soluble, assembled and active proteases by replacing the insoluble transmembrane domain with a soluble analogue. In combination with a series of solution biochemical assays, we demonstrate that the human mitochondrial AAA protease, YME1L, is a capable of mechanically unfolding stable protein substrates, requires substrates to contain a recognition sequence of defined sequence, and consumes substrates by processive unfolding and translocation. Together these results clarify a number of open questions in the field and provide a framework for the study of the wider family of membrane‐anchored AAA+ molecular machines.This research was supported by Stony Brook University.
Dynein Activity Induced by a Kunitz‐type Molecule Acting on the Proteasome
Authors: Pacheco Mario , Sciani Juliana , Morais Katia , Andrade Sonia , Chudzinski‐Tavassi Ana Marisa 
Published: June 2021

MethodsCell culture: Mia PaCa‐2 and SK‐Mel‐28 tumor cells. Western blotting: Proteins were analyzed via chemiluminescence. Confocal microscopy: Live or fixed cells were analyzed using labeled Amblyomin‐X or specific antibodies. Proteasome assay: Trypsin and chymotrypsin‐like activities were accessed using chromogenic substrates. Co‐IP: Cell lysates were submitted to Co‐IP and injected into a mass spectrometer.ResultsProteasome inhibition by Amblyomin‐X and its cellular uptake was abolished when the cells were pretreated with a dynein inhibitor. Amblyomin‐X increased Rab11A protein levels and induced its co‐localization with dynein. Several proteins were found bound to dynein, such as receptors, signaling transducers, early and recycling endosome components, proteasome, aggresome, ubiquitin, cell cycle, apoptosis and extracellular matrix components.ConclusionsDynein was required for Amblyomin‐X uptake and may transport the endosome towards the endocytic recycling compartment, because Rab11A was overexpressed, co‐localized with dynein; and Rab11A and Rab20 but not late endosome markers were bound to dynein. In addition, dynein was required for proteasome inhibition induced by Amblyomin‐X, thus suggesting its transportation to the molecular target. Some proteins, such as Bag3 and Raptor were found bound to dynein and this data corroborates with the aggresome formation and autophagy inhibition induced by Amblyomin‐X, as previously reported. Furthermore, dynein ligands indicated a possible involvement of the lipid rafts, intracellular pathways, transcription factors and membrane receptors. Those findings distinguish this molecule from proteasome inhibitors, such as bortezomib and carlfizomib. Supported by FAPESP.
Structural Basis of HIV‐1 Genome Recognition via Viral RNA‐Protein Interaction
Authors: Florwick Alyssa , Carter Greg , Keane Sarah , Summers Michael 
Published: June 2021

According to the World Health Organization, more than 35 million people worldwide are currently living with Human Immunodeficiency Virus (HIV). There are 31 antiretroviral drugs to date approved for treatment, yet there is no cure for infection. To inform new drug development strategies, we study the mechanism of genome packaging, a crucial step in viral proliferation. Packaging is mediated via the highly specific selection of the dimeric genome by the nucleocapsid (NC) domain of viral Gag polyprotein. Previous studies have identified the Core Encapsidation Signal (CES), the minimal area of the 5' untranslated region required to direct genome packaging. Using NMR spectroscopy, we have determined the 3‐D structure of CES. This structure contains tandem three‐way junctions that sequester the splice donor site. We propose that the tandem three‐way junctions, along with the psi hairpin, both of which containing a number of exposed or weakly base‐paired guanosines, have a large number of NC binding sites. To identify NC binding sites, we used site‐directed mutagenesis to mutate specific guanosines (G) to adenosines (A), which should inhibit NC binding. Using isothermal titration calorimetry, we performed binding studies with RNA constructs containing clusters of G to A mutations at the Psi hairpin and junctions. These data confirm the existence of NC binding sites at junction and hairpin Gs. Understanding the determinants of RNA‐protein interaction between CES and NC further elucidates the mechanism of genome selection. Better understanding of the structural mechanism of HIV‐1 genome recognition provides new possibilities for late‐phase viral inhibition.This research was funded by NIH/NIGMS grant 1P50GM103297.
A Cluster RCT Evaluating the Effect of Iodized Salt on Infant Development in Amhara Region of Ethiopia
Authors: Mohammed Husein , Marquis Grace , Aboud Frances , Bougma Karim , Samuel Aregash 
Published: June 2021

Iodized has been associated with the physical and mental development of children. We examined the effects of iodized salt on the growth and mental development of infant. Forty‐four communities in the Amhara region were randomly assigned and had access to iodized salt early (intervention) or later through market forces (control). A total of 1220 pregnant women were recruited and their health and micronutrient status were assessed at enrollment. Data were collected on infants (2 to 13mo), including SES, diet, anthropometry, Hb, home stimulation and mental development (Bayley scales, Cognition, Expressive language, Receptive language and Fine motor scales). The median UI excretion was higher in the intervention vs controls, for both mothers during pregnancy (161 vs 121 µg/L, p<0.0001) and infants (141 vs 110 µg/L, p<0.02). Additionally, (45 vs 60%, p<0.05) of mothers and (37 vs 44%, p<0.05) of infants were iodine deficient. The intervention had a tendency to improve cognition with an effect size of 0.09, p=0.06, but did not affect the scores for the other three bayley scales. The intervention significantly improved home score (delta=0.45, p=0.04). Iodized salt intake improved iodine status of both pregnant women and their infants, and could lead to improved infant development.Funded by Micronutrients Initiative, Canada
Plant‐Based Foods: Adolescents' Perceptions and the Consequent Implications for Policy and Practice
Authors: Ensaff H , Coan S , Sahota P , Braybrook D , Akter H , McLeod H 
Published: June 2021

A plant‐based diet, with an emphasis on plant foods and limited amounts of refined processed foods and animal products, conveys substantial health benefits. The objective of this study was to explore adolescents' attitudes and perceptions towards plant‐based foods, both in and out of the school environment.Semi‐structured focus group interviews were conducted with Year 10 students (age 14‐15 years) (n=29) attending a large secondary school in the city of Leeds, UK. The focus group interviews were audio recorded, transcribed verbatim, and categories and concepts were derived using a grounded theory methodology. Emergent theories were tested against the data collected and existing theories, until the final categories and concepts, as well as their interrelationships were well defined.Data analysis provided four main categories (Food choice parameters; Perceived drivers and benefits of plant‐based foods; Environmental food cues; Barriers to plant‐based food choice) each with their related concepts. The disconnect between plant‐based foods and adolescents' food choice parameters is highlighted, and barriers to adolescents adopting a plant‐based diet are differentiated and considered with respect to practice and policy. Opportunities to remodel and re‐present plant‐based foods in order to shift adolescents' diets are explored.The grounded theory that emerged focuses on the disconnect between adolescents' food choice parameters and plant‐based foods. The study suggests implications for practice and policy with opportunities to improve and manage adolescents' diet.Grant Funding Source: Alpro Foundation
Determine Entry‐Level Registered Dietitian Nutritionist's Practices in and Barriers to Participation in Advocacy and Nutrition‐Related Policies
Authors: Mondelli Virginia , Sowa Diane , Foley Sharon , Chen Yimin 
Published: June 2021

BackgroundThere is limited information regarding Registered Dietitian Nutritionist's (RDN) motivation and barriers to participation in advocacy and public policy‐related activities. The purpose of this study was to determine entry‐level (less than or equal to three years of experience) RDN's participation in advocacy for nutrition‐related policy.MethodsIn a cross‐sectional nation‐wide study, 3,164 (22% Response Rate) entry‐level RDNs responded to an online questionnaire via SurveyMonkey.com about their motivations and barriers to participating in advocacy. Descriptive statistics and chi square tests were utilized. The study had IRB approval.ResultsIn this largely female (95%) sample of young adult (80% aged 20 to 30 years old) RDNs, 51% were motivated to participate in public policy activities. RDNs indicated they were motivated to participate in advocacy in order to enhance the health of the nation (99%) and because they were passionate about nutrition policy (99%). Of those RDNs who were not motivated (49%), a lack of time (37%) and interest (27%) were the main reasons given for their lack of motivation. Respondents noted lack of time (83%), role models (71%) and knowledge on “how to” participate (71%) as major barriers to participation of advocacy.ConclusionsOver fifty percent of entry‐level RDNs were motivated to participate in advocacy. To overcome the barriers in getting RDNs involved in public policy more education about “how to” advocate and spotlighting advocate role models is needed to improve RDN's participation in advocacy.
Correlation macro nutrient intake with body fat among adolescent gymnastic, in East Java Province, Indonesia
Authors: Adiningsih Sri , Indrawati Desi 
Published: June 2021

Body fat is the biggest source of energy for athletes to perform exercise. The Coach strict intake macronutrient to get low body weight for gymnast. Gymnast suffer energy deficit which affect their body fat because of high energy expenditure during exercise and food strict.AimsThis research was to analyze correlation between macronutrient intake with body fat of adolescent gymnast.MethodsThis observasional research with using cross sectional analytic, which used entire population as sample which consisted of 9 male gymnast and 14 female gymnast below 19 age of Gymnastic athlete club east java province, Indonesia. By using 24 hour's recall for measured food intakes and BIA for measured body composition.ResultTable 1. Character Physically of Sample

Character
Male (n=9)
Female (n=14)
Total (n=23)

Age (year)
12.4 ± 1.9
14.0 ± 2.2
13.39 ± 2.21

Height (cm)
140.7 ± 12.8
145.9 ± 8.8
143.9 ± 10.6

Weight (kg)
35.4 ± 8.5
38.3 ± 7.5
37.2 ± 7.8

IMT (km/m2)
17.6 ± 1.8
17.8 ± 2.1
17.7 ± 1.9

IMT/age
normal
normal
‐

Body fat (%)
8.9 ± 3.9
16.2 ± 5.0
13.3

10‐12 year
9.3 ± 2.8
13.3 ± 2.4
11.3

13‐15 year
6.4 ± 1.3
15.6 ± 5.1
11.9

16‐18 year
17 ± 0
20.1 ± 5.3
19.4

Male gymnast had 8,9% average body fat and female gymnast had 16,2% average body fat.

Intakes of macronutrient
Male
Female
ρ‐value

average ± SD
RDA
average ± SD
RDA
male
female

Carbohidrate (gram)

10‐12 year
201 ± 19.1
289
84.2 ± 25.3
275
0.018
0.138

13‐15 year
181.8 ± 12.7
340
87.1 ± 40.7
292

16‐18 year
267.1 ± 0
368
99.1 ± 15.3
292

Protein (gram)

10‐12 year
58.9 ± 6.2
56
29.3 ± 8.9
60
0.025
0.035

13‐15 year
49.5 ± 7.4
72
27.6 ± 6.9
69

16‐18 year
71.9 ± 0
66
35.7 ± 5.9
59

Lemak (gram)

10‐12 year
57.9 ± 6.6
70
28.4 ± 9.1
67
0.002
0.059

13‐15 year
48.0 ± 2.7
83
29.4 ± 12.7
71

16‐18 year
63.2 ± 0
89
31.0 ± 4.7
71

Table 2. Crosstab Macronutrient Intake with Body FatShowed body fat percentage had correlation with carbohidrat intake (p=0.018, r=0.758), protein intake (p=0.025, r=0.732), and fat intake (p=0.002, r=0.881) of male gymnast. Only protein intake (p=0.035, r=0.565) had correlation with body fat percetage in female gymnast, whereas carbohidrate (p=0.138) and fat intake (p=0.059) had no correlation with body fat percentages.ConclusionGymnast need digital scales BIA to control body fat percentages. Decrease body fat should make gymnast improve physical fitness and performance of gymnast.