bluuebunny/crossref_metadata_2025_split huggingface dataset:
Zoanthus sociatus extract blocks the nifedipine sensitive‐Ca+2 influx in beta cells and impairs glucose stimulated insulin secretion
Authors: Diaz-Garcia Carlos Manlio , Sanchez-Soto Carmen , Fuentes-Silva Deyanira , Leon-Pinzon Carolina , Dominguez-Perez Dany , Varela Carlos , Rodriguez-Romero Adela , Castañeda Olga , Hiriart Marcia 
Published: June 2021

Crude extracts from coelenterates have been a source of neurotoxins that still possesses a great potential in the discovery of novel drugs. Studies about the effect of cnidarians venoms upon insulin release are not common, despite the coincidences in ionic mechanisms of glucose stimulated insulin secretion and neurotransmitter release. In the present work we tested the effect of a crude extract of Zoanthus sociatus on insulin secretion in isolated rat β‐cells and its toxic effects in vivo.We observed that 50 ug/ml of the extract inhibited insulin release in a 40 % when incubated at 15.6 mM glucose and that a low molecular weight fraction was responsible of this effect. Moreover, the active fraction impaired intracellular Ca+2 rises in response to high glucose and high K+‐induced depolarization. The results suggest the presence of voltage‐gated Ca+2 channels inhibitors in the active fraction, which cause glucose intolerance when rats are intraperitoneally inoculated with the extract.This work was supported by Gobierno del Distrito Federal PICDS08‐72, CONACYT CB2009‐131647, and DGAPA‐PAPIIT IN215611, Universidad Nacional Autónoma de México. Carlos Manlio Díaz‐García was supported by a doctoral fellowship from Consejo Nacional de Ciencia y Tecnología (CONACyT).
Protein Structure Initiative:Biology‐Materials Repository: Developing a Public Resource for Structural Biology Plasmids
Authors: Cormier Catherine , Park Jin , Fiacco Michael , Steel Jason , Kramer Jason , LaBaer Joshua 
Published: June 2021

The Protein Structure Initiative: Biology‐Materials Repository (MR; http://psimr.asu.edu) is a centralized repository that sequence‐verifies, store and distributes plasmids created as part of the high throughput structural genomics efforts of the PSI. Each plasmid is fully sequence‐verified and stored, along with plasmid annotation such as the full length sequence, vector information, and associated publications, in a freely available, searchable database called DNASU (http://dnasu.asu.edu). Each plasmid links to external resources, including the PSI Structural Biology Knowledgebase (http://sbkb.org) facilitating cross‐referencing of plasmids to additional protein annotations and experimental data. Recent developments to DNASU include automation of our Expedited Material Transfer Agreement (MTA) process, which eliminates delays caused by MTA processing. As of November 2011, over 50,000 protein expression plasmids and 85 empty vectors from the PSI are available upon request from DNASU. By providing expression‐ready plasmids to the research community, we are facilitating the study of the biological function of proteins whose structures have been solved by the PSI.This work was supported by the National Institute of General Medical Sciences.
Isolation and mass spectrometric analysis of native protein complexes in rat liver mitochondrial contact sites
Authors: Lee Kwangwon , Kerner Janos , Hoppel Charles 
Published: June 2021

Contact sites (CS) are structures where the mitochondrial inner and outer membrane are fused and can be isolated by discontinuous sucrose gradient centrifugation of crude outer membrane derived from swollen/shrunk rat liver mitochondria. We used a combination of Blue‐Native PAGE (BNE) and LC‐MS/MS to isolate and characterize the protein complexes present in CS.CS proteins were extracted with Triton X‐100 and separated into eleven different size bands by BNE. All eight bands, analyzed by LC‐MS/MS, contained voltage‐dependent anion channel (three isoforms), long‐chain acyl‐CoA synthetase 1, phosphate carrier. Four bands between 600 kDa and 960 kDa contained carnitine palmitoyltransferase 1a, very long‐chain acyl‐CoA dehydrogenase, and the trifunctional protein (both Î± and Î2). In the same four bands, adenine nucleotide translocase (two isoforms) and apoptosis‐inducing factor 1 were identified. Of interest, carnitine‐acylcarnitine translocase was found only in the bands between 140 kDa and 400 kDa. Based on the proteins identified, liver mitochondrial CS from fed rats contain protein complexes of various high molecular weight.We propose these protein complexes of various sizes represent carnitine transport system as well as very long chain fatty acid oxidation enzymes, nucleotide transport system and apoptosis system.Supported by the NIH grant DK‐066107
A novel RNA detection technology for gene expression analysis in living cells
Authors: Weldon Don S , Ko Alex , Johnston Grace , Hsu Matthew 
Published: June 2021

Detecting gene expression has traditionally been limited to technologies that examine expression in lysed or fixed cell populations. The ability to detect gene expression in live cells would allow for more physiologically relevant information based on a cell's response to given stimuli. Determining which genes were up or down regulated in those cells provides insight into complex gene regulatory networks and cell function. Here we present a novel RNA expression detection technology capable of detecting specific mRNA and miRNA in live, intact cells. This technology allows for carrier‐free cellular uptake of the reagent, followed by detection of target RNA, with the ability to perform downstream analysis in the same sample. This reagent has no toxic effects on cells, and we have demonstrated the ability to isolate and further propagate live cell populations based on gene expression level. This would allow researchers to detect RNA and subsequently protein levels within the same cell.Since current methods to interrogate the cellular RNA examine non‐native, amplified RNA targets, the relevance of such data often does not correlate to in vivo results. Furthermore, these methods do not provide RNA information from individual cells, and cells probed by these methods cannot be further studied following detection. Our technology presented here leaves the cells unchanged following detection, and therefore this same cell population can be used for downstream experiments. This research was funded by EMD Millipore.
Rhodopsin Interacts With Retinal Guanylate Cyclase; Involvement In Phototransduction
Authors: Bondarenko Vladimir 
Published: June 2021

We expressed bovine Rho and intradiscal domain of retGC in HEK293 and used membrane and soluble fractions pull down assay to demonstrate interaction between these proteins. We also co‐expressed fluorescently‐tagged proteins in cultured cells to analyze their co‐localization in cell.It has been previously shown that retGC activity in light‐exposed rod outer segment membranes was much higher than activity of the same membranes isolated under dark conditions suggesting that communication of retGC with activated rhodopsin is required for its full activation by GCAP. To study interaction between Rho and retGC we used pull down assay and colocalization experiments. Fluorescent protein‐tagged bovine Rho and intradiscal domain of retGC were expressed in HEK293 cells and membrane fraction containing Rho and soluble fraction containing intracellular domain of retGC were prepared. Membrane and soluble fractions were mixed together and upon incubation, membranes were separated from soluble fraction by centrifugation and the presence of both recombinant proteins in washed membranes has been analyzed by western blot. Intradiscal domain of retGC was present only in cell membranes containing Rho suggesting direct interaction between these proteins, which was also supported by colocalization experiments.
KCS1 deletion in Saccharomyces cerevisiae leads to a defect in translocation of autophagic proteins and reduces autophagosome formation
Authors: Chen Michael Po-Hao , Taylor Robert , Chou Chia-Ching , Jin Shengkan 
Published: June 2021

Inositol phosphates are implicated in the regulation of autophagy; however, the exact role of each inositol phosphate species is unclear. In this study, we systematically analyzed the highly conserved inositol polyphosphate synthesis pathway in S. cerevisiae for its role in regulating autophagy. Using yeast mutants that harbored a deletion in each of the genes within the inositol polyphosphate synthesis pathway, we found that deletion of KCS1, and to a lesser degree IPK2, led to defect in autophagy. KCS1 encodes an inositol hexakisphosphate/heptakisposphate kinase that synthesizes 5‐IP7 and IP8; and IPK2 encodes an inositol polyphosphate multikinase required for synthesis of IP4 and IP5. We characterized the kcs1Δ mutant strain in detail. The kcs1Δ yeast exhibited reduced autophagic flux, which might be caused by both the reduction in autophagosome number and autophagosome size as observed under nitrogen starvation. The autophagy defect in kcs1Δ strain was associated with mislocalization of pre‐autophagosomal structure (PAS), a defect in Atg18p release from the vacuole membrane and mislocalization of PtdIns(3)P under nitrogen deprivation conditions. Interestingly, formation of autophagosome‐like vesicles was commonly observed to originate from the plasma membrane in the kcs1Δ deletion strain. Our results indicate that lack of KCS1 interferes with proper localization of some autophagic proteins, leads to reduction of autophagosome formation, and causes the formation of autophagosome‐like structure in abnormal subcellular locations.Michael Po‐Hao Chen and Robert Taylor Jr. both contributed equally to the work.
RTK Phosphorylation
Authors: Schmidt Jane , Schmidt Jane 
Published: June 2021

A homogeneous multiplex immunoassay for RTK phosphorylation state that is adaptable to HTSJane Schmidt, Ph.D., Judy Haning, Shaya Anderson, Richard Fuerstenberg. R&D Systems, Inc. Minneapolis, MNReceptor Tyrosine Kinases (RTKs) are widely expressed transmembrane protein receptors for various extracellular signaling molecules. Binding of a specific ligand leads to phosphorylation at specific sites in the cytoplasmic tail which triggers further signal transduction processes. RTKs play critical roles in the regulation of developmental processes including cell survival, proliferation, and motility, which are involved in cancer formation when not properly regulated. For these reasons, RTKs have become popular therapeutic targets for drug development. The Human RTK VersaMAP Magnetic Multiplex System kits are bead‐based multiplex immunoassays for the analysis of RTK phosphorylation. Due to the homogeneous assay format and small sample volume, these assays can be readily adapted for high‐throughput screening with minimal manual intervention from cell treatment to data collection. To demonstrate this, several different cell lines were grown in 96‐well plates, treated with various stimulants or inhibitors, and lysed in the same plate. Diluted lysates were transferred to the assay plate for analysis of RTK phosphorylation by Human RTK VersaMAP assays in two multiplexes of 25 each. Using these assays, relative phosphorylation states were successfully determined for the various cells and treatments.
Student Learning Outcomes and Attitudes when Biotechnology Lab Partners are of Different Academic Levels
Authors: Witherow D. Scott , Miller Heather , Carson Susan 
Published: June 2021

Courses teaching fundamental concepts and methodologies in biochemistry and molecular biology can be beneficial for both undergraduate and graduate students alike. Should these courses be taught differently and separately to students of different levels? Anecdotal evidence has shown that students prefer pairing with someone of the same academic level. However, retention of main ideas in a peer learning environment has been shown to be greater when partners have dissimilar abilities. In our “Manipulation and Expression of Recombinant DNA” course, which can be taught as an advanced Biochemistry/Molecular Biology course, we tested the hypothesis that there will be enhanced student learning when lab partners are of different academic levels. Learning outcomes were met by both undergraduate and graduate students, regardless of pairing. Average undergraduate grades on every assessment method increased when paired with graduate students. Many of the average graduate student grades also increased modestly when paired with an undergraduate. Attitudes toward working with partners dramatically shifted. The work presented here suggests that offering dual‐level courses benefit students of all levels. This format is useful for institutions that wish to offer “boutique” courses where student enrollment may be low, but specialized equipment and faculty expertise is needed.This work was supported by the NCSU Biotechnology Program.
Designing and Implementing Team‐Based Learning Exercises in a Medical Biochemistry and Molecular Biology Course
Authors: McKee Edward E. 
Published: June 2021

ContextTeam‐Based Learning is a well‐characterized and highly interactive pedagogy that has been used for some time in a variety of disciplines. However, its use in medical education and in Medical Biochemistry is more recent. The popularity of Team‐ Based learning appears to be growing as medical education strives to become more interactive, particularly because TBL is an interactive teaching strategy that can be delivered by a single faculty member. I have made extensive use of TBL for the past 5 years in my course in Medical Biochemistry and Molecular Biology.ObjectivesThe poster will describe the basics of TBL and how TBL has been used in my course to integrate and apply content. The added value of TBL in a competency based curriculum is also described. TBL is a basis for students to practice, refine, and assess their peers on a variety of competencies required for team interaction, including: communication, life‐long learning, self awareness, ethical and moral reasoning, problem solving, and professionalism.ConclusionThe TBL sessions in this course were highly valued by the students and provided a highly interactive environment for learning content and perfecting team skills.
Contribution of Unilever Reformulation to Changes in Nutrient Intakes: Modeling of Sodium & SAFA
Authors: Roodenburg Annet Johanna Christina , Dotsch-Klerk Mariska 
Published: June 2021

Based on the WHO Strategy for Diet Physical Activity and Health (2004), the food industry has been reformulating products into a healthy direction. In 2005, Unilever started as one of the first global food companies with an active programme aimed at food reformulation; now continued under the Unilever Sustainable Living Plan.To estimate the effect of reformulation by Unilever of specific foods in the period 2005–2007, the Monte Carlo Risk Assessment model was used. Inputs for this model were food intakes from surveys and nutrient composition of foods (NEVO) as well as data from Unilever (including market shares).ResultsSodium intake from soups was reduced 7%–19%. In absolute numbers this was 10–20 mg sodium: representing <1% of total daily sodium consumption (> 2400 mg/d) for the total population.SAFA intake from spreads and oils was reduced 12–33%. This was 0.6–1.4 g/d per day: 3–4% of total daily SAFA consumption (32g/d) for the total population.The analyses show changes in nutrient intakes due to reformulations within product groups. The public health relevance depends on the relative importance of these product groups in the total food consumption pattern of a population.
Bioaccesibility of Carotenoids from Maize Flour with Varying Levels of Resistant Starch Type 2 and 3
Authors: Surles Rebecca Lynn , Tanumihardjo Sherry , Palacios Natalia 
Published: June 2021

BackgroundIt has been proposed that resistant starch may decrease the bioavailability of provitamin A carotenoids from some foods possibly by interfering with micelle formation in the gut. Research is needed to determine if the type of resistant starch (RS) and processing methods of food like maize influence bioavailability of carotenoids. To do this it is first necessary to assess interactions between provitamin A carotenoids and resistant starch content.ObjectThree Stage In Vitro Digestion Method will be used to assess micellarization efficiency of carotenoids from maize flour with various concentrations of resistant starch to determine bioaccesibility of provitamin A.Study designType of RS in the maize flour depends on the processing method. Uncooked flour will have higher proportions of resistant starch 2 (RS2) whereas cooked and cooled maize flour will be higher in the retrograded starch or resistant starch 3 (RS3). Starch in cooked maize will be gelatinized increasing the availability of starch (AS). Total RS will be analyzed with a Megazyme kit and carotenoids will be quantified before and after in vitro digestion in maize flour with various carotenoid and RS concentrations. Total starch and oil will be quantified to assess bioaccesibility interactions. Results from this study will provide necessary research towards understanding provitamin A bioaccesibility in maize. Supported by Harvest Plus.
*All the following procedures will be conducted in the cooked and raw samplesimage*All the following procedures will be conducted in the cooked and raw samples
The MTHFR C677T gene polymorphism controversy. A protective effect for renal function in patients without hyperomocysteinemia under adequate alimentary regimen
Authors: Trovato Guglielmo M , Catalano Daniela , Ragusa Angela , Trovato Francesca M. , Pirri Clara , Di Nora Concetta , Buccheri Maria Antonietta , Tonzuso Antonia , Martines G. Fabio 
Published: June 2021

The association with atherosclerosis, renal failure and Cardio‐Vascular events of MTHFR gene C677T polymorphism is still controversial.Aimto investigate relationship of MTHFR 677C>T polymorphisms with Renal Function in non‐diabetic subjects without hyperhomocysteinemia. We studied 178 subject: 78 MTHFR C677CC (no polimorphism), 52 HFR C677CT and 48 MTHFR C677TT (with polymorphism) patients (age 50.93±14.85 yrs, BMI 27.95±5.98 kg/m2), all with adequate adherence to Mediterranean Diet Italian Guidelines and normal serum homocysteine. Glomerular Filtration Rate (eGFR), Insulin Resistance (HOMA), Parathyroid Hormone, UltraSound Doppler Renal Resistive Index of renal artery and Echocardiography Left Ventricular Mass/m2 were assessed.ResultsMTHFR 677C>T polymorphism has higher eGFR (73.8±27.99 vs. 58.64±29.95; p<0.001) and lower Renal Failure risk (OR 0.684; CI 95% 0.313–1.494) in comparison with controls. By multivariate analysis (MLR), MTHFR polymorphism favorable effect is independent from the negative effects of insulin resistance, hyper‐parathyroidism and obesity. Conclusiono. MTHFR 677C>T gene polymorphism is associated with better renal function in well‐nourished patients, independently from other variables and homocysteine levels.
Hass Avocado Improves Postprandial Vascular Reactivity When Consumed in Combination with Cooked Hamburger Meat
Authors: Li Zhaoping , Zhang Yanjun , Henning Susanne M. , Wong Angela , Jones Alexis , Zerlin Alona , Thames Gail , Bowerman Susan , Heber David 
Published: June 2021

The Hass Avocado contains monounsaturated fats (oleic acid) and antioxidants (carotenoids, tocopherol). Cooked hamburger meat contains lipid peroxidation products formed during cooking. After ingestion, the stomach functions as a bioreactor, generating lipid peroxides and this process can be inhibited when antioxidants are ingested together with the meat. The present study was conducted in eleven healthy subjects on two separate occasions consuming either a 250 gram hamburger patty alone (ca. 436 Cal and 25 g fat) or the hamburger together with 68 grams of avocado flesh (an additional 114 Cal and 11 g of fat for a total of 550 Cal and 36 g fat), a common culinary combination, to assess effects on vascular health. Following a standard peripheral arterial tonometry (PAT) method (Itamar) to calculate PAT index there was a significant decrease 2 hours following hamburger ingestion alone (2.19 ± 0.36 vs. 1.56 + 0.21, p=0.0007) which was prevented when avocado was ingested with the burger (2.17 ± 0.57 vs. 2.08 ± 0.51, NS (p= 0.68). Monocytes were isolated from postprandial blood samples and I kappa‐B alpha (IKBa) protein measured to assess effects on activation of inflammation. At 3 hours, there was a significant preservation of IkBa (131 % vs. 58 %, p=0.03) consistent with inhibition of inflammation via the NF‐kappa B pathway. Triglycerides, MDA, IL‐6, and TNF‐alpha increased postprandially but were not further increased when avocado was ingested with the burger compared to burger alone, despite the added fat and calories from the avocado. These observations are consistent with beneficial antioxidant and vascular health effects of ingesting added Hass avocado with a hamburger patty.Study Supported by The Hass Avocado Board
A high fat/high fructose (HF) diet does not exacerbate the metabolic or cardiac functional consequences induced by diabetes in C57Bl6J mice
Authors: Lachance Dominic , Vaillant Fanny , Shi Yanfen , Daneault Caroline , Frayne-Robillard Isabelle , Deschepper Christian , Carpentier Andre , Tardif Jean-Claude , Rosiers Christine Des 
Published: June 2021

Background & ObjectivesHigh fat or high sugar diets are associated with increased risk for coronary heart disease but have not been found to exacerbate cardiac dysfunction in rodent models of heart failure. We thus tested the effect of HF diet in a mouse model of diabetes and its associated cardiac dysfunction.MethodsFive‐week‐old C57Bl6J male mice received either standard (Std) or HF diet for 2 weeks, then received multiple injections of either low doses (40 mg/kg) of streptozotocin (STZ) or vehicle (Veh).ResultsTen weeks after STZ injections (and compared to Std‐Veh mice), both STZ groups displayed similar (i) fasting plasma glucose and insulin levels, which were respectively increased and decreased (p<0.001) and (ii) cardiac systolic dysfunction, as evidenced by a 8% lower ejection fraction (p<0.05) at echocardiography. However, the increase in plasma levels of ketone bodies, glycated haemoglobin and branched chain amino acids was smaller (p<0.05) in HF‐STZ mice than in Std‐STZ at both 6 and 12 weeks. Likewise, HF‐STZ mice maintained (in contrast to Std‐STZ mice) body weight and epidydymal fat mass as well as Std‐Veh.ConclusionNot only did HF not exacerbate diabetes‐induced cardiac dysfunction, but it even attenuated the systemic metabolic alterations induced by STZ, which may in the long term delay the development of heart failure. (Supported by CIHR & HSFC)
Combining radiology technicians and medical students to develop imaging and interpretation skills
Authors: Aabo Megan , Delamarter Taylor , Benninger Brion 
Published: June 2021

INTRODUCTIONFirst year medical students (MS1) are generally introduced to radiology during their anatomy course with a brief overview and limited roentograph teaching. Imaging interpretation skills could be taught during the anatomy dissection lab (ADL). In a vertical curriculum, it is essential to integrate basic and clinical sciences to enable MS1 to confidently apply critical thinking skills. The objective of this study was to combine MS1 with radiology technicians (RTs) during ADL to develop and improve MS1 imaging and interprtation skills.METHODSLiterature search was conducted on combining MS1 and RTs during ADL. Exam scores were analyzed and a questionnaire was conducted for assessment.RESULTSLiterature search revealed no studies combing MS1 and RTs during ADL course. Exams and a questionnaire revealed both an objective and perceptual success.DISCUSSIONAn interprofessional educational model was designed to expose MS1 to various radiologic mediums that focused on fundamentals of capturing desired images. RT's taught MS1 the how, what, where and why to capture and interpret imaging. Exam scores were high and a questionnaire suggested this novel model an educational success.CONCLUSIONThis pilot study introduced a novel interprofessional education model to develop and improve imaging knowledge during an anatomy course within a vertical curriculum.