bluuebunny/crossref_metadata_2025 huggingface dataset:
The Impact of AB5 Toxin Binding on Ganglioside Mimicry in Campylobacter jejuni Lipooligosaccharides
Authors: Patry Robert T , Stahl Martin , Sacher Jessica , Vallance Bruce A , Szymanski Christine M
Published: November 2023

Campylobacter jejuni is an important pathogen in the developed world, causing more cases of acute gastroenteritis than any other bacteria. C. jejuni is also the leading cause of paralysis in the world, causing Guillain‐Barré syndrome in humans by mimicry of nerve gangliosides with its surface exposed lipooligosaccharides (LOS). In some cases this mimicry elicits a breakdown in immune tolerance and results in self‐reactive antibodies which damage the nerve cells. Interestingly, the commonly mimicked GM1 ganglioside is also the main receptor for cholera toxin (CT) from Vibrio cholerae as well as heat‐labile enterotoxin (LT) from Escherichia coli. We have shown that these two toxins bind C. jejuni strains that mimic GM1 ganglioside and clear bacterial growth. The binding and clearing phenotypes were observed closely using transmission and scanning electron microscopy. C. jejuni 11168 shows natural variation in its ability to mimic GM1 gangliosides. Due to phase‐variation in cgtB encoding the galactosyltransferase, this strain exists as a mixture cells with and without GM1 mimics. It was unclear why the strain retained this variation, given that mimicry of host gangliosides lends a significant fitness advantage by camouflaging the bacterium from host immune defenses. Therefore, LOS was isolated from 11168 exposed to CT and LT toxin and compared to the unexposed by silver staining and far Western blotting. These experiments revealed the loss of a band from toxin exposed LOS and a marked decrease in the ability to bind toxin. Furthermore, when genomic DNA was isolated from these two groups and the cgtB gene was sequenced, the variation that existed in the unexposed group was no longer observed in the presence of the toxins. This led to the hypothesis that C. jejuni 11168 may retain variation in the cgtB gene in order to avoid negative effects upon contact with these toxins in the host or the environment. In addition, when we used a strain of E. coli engineered to display the GM1 mimic on its surface, no clearance was observed suggesting that this interaction is pathogen specific. Our study identifies a new interaction between C. jejuni and the AB5 toxins that has not been previously described and provides a possible new relationship within the intestinal microbiome.

Support or Funding Information

Funding provided by the Natural Sciences and Engineering Research Council of Canada, Alberta Innovates Technology Futures and Alberta Innovates Health Solutions.
The Antibiotic Activity of Tea Tree, Eucalyptus, and Cinnamon Bark Oils
Authors: Rohraff Dallas , Morgan Roderick
Published: November 2023

The emergence of antibiotic resistant bacteria is of pressing concern, especially given that the pharmaceutical industry has decreased funding for infectious disease drug discovery programs. It is estimated that over 2 million people are infected with antibiotic resistant strains of bacteria each year, resulting in over 23,000 deaths. Researchers are currently looking for novel antibiotics in alternative sources. Essential oils, derived from plants, are an ancient homeopathic remedy that are traditionally known to have antimicrobial properties. Therefore, several essential oils were examined in search of novel antibiotics. Tea tree (Melaleuca alternifela), eucalyptus (Eucalyptus globulus), and cinnamon bark (Cinnamomum cassia Blume) oils showed antibiotic activity in Kirby‐Bauer disk diffusion assays. The Minimum Inhibitory Concentration (MIC) values for tea tree, eucalyptus, and cinnamon bark oils were determined to be 1.25%, 0.313%, and 0.0391%, respectively. All three oils proved effective against both Gram‐positive Staphylococcus aureus and Gram‐negative Escherichia coli. Cinnamon bark oil was the most effective and was further characterized. Cinnamon bark oil retained its MIC in the presence of 10% human serum protein, indicating that it does not interact with proteins in the bloodstream. Cinnamon bark oil showed bactericidal activity in a time kill assay. To determine the active component within cinnamon bark oil, an analysis of its constituents was conducted. Three of the main components of cinnamon bark oil, trans‐cinnamaldehyde, cinnamyl cinnamate, and benzyl cinnamate, were subjected to preliminary testing via Kirby‐Bauer disk diffusion tests. Trans‐cinnamaldehyde appeared to be the only effective component against both S. aureus and E. coli. Results suggest that cinnamon bark oil may contain a promising novel antibiotic.

Support or Funding Information

This research was supported by Grand Valley State University's Student Summer Scholars program.
Decreased Insulin Signaling Causes Loss of Cardiac PFK‐2 in Diabetes
Authors: Bockus Lee , Humphries Kenneth
Published: November 2023

Cardiac 6‐phosphofructo‐2‐kinase/fructose‐2,6‐bisphosphatase (PFK‐2; PFKFB2), a central metabolic enzyme that regulates glycolysis in the heart, is regulated by insulin and adrenergic signaling. Although phosphorylation is known to increase its kinase activity, it is unknown whether insulin signaling and diabetes also affects its content and thus its active capacity. This is critical to know because PFK‐2 content can have a significant impact on cardiac metabolic flexibility. We therefore employed multiple mouse and primary cardiomyocyte models with deficient insulin signaling. In streptozotocin‐induced type I diabetic mice we found a 70% decrease in PFK‐2 protein content. Further, the remaining PFK‐2 is not phosphorylated by activation of PKA in vivo. A reduction in cardiac PFK‐2 content also occurs with overnight fasting and a high fat diet mouse model of type II diabetes. By metabolomic analysis, fructose‐6‐phosphate is elevated 3‐fold with 24h fasting and 3‐phosphoglycerate is reduced 50%, suggesting glycolysis is blocked at phosphofructokinase‐1. No difference was found in AMP levels or citrate, indicating that the reduced content and activity of PFK‐2 is a primary reason glycolysis is impeded in the fasting state. The loss of PFK‐2 can be recapitulated in adult mouse cardiomyocytes by the removal of insulin from media or supplementation with lipids, both of which reduce Akt‐mediated phosphorylation. Inhibition of insulin signaling in cardiomyocytes by wortmannin, a PI3K inhibitor, causes a marked loss of PFK‐2 with a half‐life of 2 hours. In summary, reduced insulin signaling in the heart in both types of diabetes and fasting results in PFK‐2 degradation and impaired glycolysis.

Support or Funding Information

Research reported in this study was supported in part by an Institutional Development Award (IDeA) from the National Institute of General Medical Sciences of the National Institutes of Health (P20GM104934), by the Oklahoma Center for the Advancement of Science and Technology (HR13‐183), and by the American Heart Association (14GRNT20510031).
A role of NMDA receptors in regulating KATP and Kv2.1 channel trafficking in pancreatic β‐cells
Authors: Chen Pei‐Chun , Wu Yi , Ruan Jing‐Syuna , Shyng Show‐Ling
Published: November 2023

Leptin secreted from adipocytes is known to act on pancreatic β‐cells directly to suppress insulin secretion. We recently showed that leptin reduces β‐cell excitability by promoting surface expression of both KATP and Kv2.1 channels in β‐cells via a signaling mechanism involving AMPK and its upstream kinase CaMKKβ. However, how leptin activates the CaMKKβ/AMPK signaling axis remains unclear. Here we tested the hypothesis that NMDA receptors, which are permeant to Ca2+, may be involved. Using the insulinoma INS‐1 cells, we show that the NMDA receptor antagonists MK‐801 or AP‐5 blocked the effect of leptin on both KATP and Kv2.1 channels; conversely, the receptor agonist NMDA alone is sufficient to recruit KATP and Kv2.1 channels to the β‐cell membrane. The effects of the NMDA receptor agonist and antagonists on KATP and Kv2.1 channels corresponded with their effects on pAMPK levels, indicating that the involvement of NMDA receptors is downstream of leptin but upstream of AMPK. Western blots confirmed that INS‐1 cells indeed express the obligatory NMDA receptor subunit GluN1. Electrophysiological recording further demonstrated the presence of NMDA evoked currents in INS‐1 and human β‐cells. Our findings suggest that NMDA receptors are important for mediating the effect of leptin to regulate trafficking of Kv2.1 and KATP channels in β‐cells.

Support or Funding Information

(Supported by NIH R01DK057699 to SLS and MOST 1032320B006005MY2 to PCC.)
Inhibitors of Ribosome Rescue Arrest Growth of Francisella tularensis at All Stages of Intracellular Replication
Authors: Goralski Tyler D.P. , Dewan Kalyan K. , Alumasa John N. , Avanzato Victoria , Place David E. , Markley Rachel L. , Katkere Bhuvana , Rabadi Seham M. , Bakshi Chandra Shekhar , Keiler Kenneth C. , Kirimanjeswara Girish S.
Published: November 2023

Francisella tularensis ssp. tularensis is a gram‐negative, facultative intracellular bacterium, which is classified as a Tier 1 Select Agent by the CDC due to its high infectivity and ease of propagation. Attempts to develop an effective vaccine have been unsuccessful, due in part to the organism's ability to suppress or bypass the host immune response early after infection. F. tularensis strains resistant to multiple antibiotics are a biowarfare threat. In the absence of an effective vaccine, new antibiotic targets and compounds are needed to ensure biodefense. Bacteria require at least one pathway to rescue ribosomes stalled at the end of mRNAs. The primary pathway for ribosome rescue is trans‐translation, which is conserved in >99% of sequenced bacterial genomes. Some bacterial species also have backup systems, such as ArfA or ArfB, which can rescue ribosomes in the absence of sufficient trans‐translation activity. Small molecule inhibitors of ribosome rescue have broad‐spectrum antimicrobial activity against bacteria grown in liquid culture. These compounds were tested against F. tularensis to determine if they can limit bacterial proliferation during infection of eukaryotic cells. The inhibitors KKL‐10 and KKL‐40 exhibited exceptional antimicrobial activity against both attenuated and fully virulent strains of F. tularensis in vitro and during ex vivo infection. Addition of KKL‐10 or KKL‐40 to macrophage or liver cells at any time after infection by F. tularensis prevented further bacterial proliferation. When macrophages were stimulated with the pro‐inflammatory cytokine interferon‐γ before being infected by F. tularensis, addition of KKL‐10 or KKL‐40 reduced intracellular bacteria by >99%, indicating that the combination of cytokine induced stress and a nonfunctional ribosome rescue pathway is fatal to F. tularensis. Neither KKL‐10 nor KKL‐40 were cytotoxic to eukaryotic cells in culture. These results demonstrate that ribosome rescue is required for F. tularensis growth at all stages of its infection cycle, and suggest that KKL‐10 and KKL‐40 will be good lead compounds for antibiotic development.

Support or Funding Information

This work was supported by NIH (AI077917 to Girish S. Kirimanjeswara and GM068720 to Kenneth C. Keiler) and CAS/PSU start up funds to Girish S. Kirimanjeswara. The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Microparticle analysis of clinical plasma samples using acoustic trapping
Authors: Bryl‐Gorecka Paulina , Sathanoori Ramasri , Gidlöf Olof , Svensson Siv , Evander Mikael , Laurell Thomas , Olde Bjorn , Erlinge David
Published: November 2023

Objective

Endothelial dysfunction, inflammation and platelet activation are conditions that predispose for development of cardiovascular disease. These are also conditions previously shown to trigger release of microparticles (MPs). As isolation of MPs from plasma is time consuming and requires large sample volumes, we decided to develop an alternative method based on acoustic trapping. Acoustic trapping is a non‐contact method where sound waves are used to retain particles in a microfluidic channel (Figure 1). Exciting a capillary with a piezoelectric transducer that creates a local acoustic standing wave field will enable trapping and clustering of particles in this zone. Submicron particles can subsequently be trapped on the larger particles by inter‐particle acoustic forces, so called “seed trapping”. As a test of the method we analyzed the change in endothelial MPs (EMPs) release that was stimulated by physical excersise in healthy controls and subjects with coronary artery disease.

Methods

Patient and control samples: 8 patients with history of coronary artery disease and 5 healthy subjects underwent physical exercise (cycling). Blood samples were collected into EDTA tubes before and 1h after exercise. The samples were centrifuged at 1,600 × g for 15 minutes and then stored at −80°C.

Isolation of MPs: Plasma samples were subjected to centrifugation (2 times 1600 × g, 15 min, 4°C) in order to obtain platelet‐free plasma. 25 ul of sample was aspirated and the microparticles were trapped and subsequently eluted in 100 ul PBS.

Staining of CD62E (E‐selectin)+ EMPs: The microparticles were stained with 3 ul of PE‐conjugated anti‐CD62E antibody for 45 min at room temperature, in the dark with constant shaking, followed by FACS analysis using a BD Accuri C6 cytometer.

Results

CD62E+ MPs ranged from 25 to 77 and 20 to 62 events/ul for patients and controls respectively. A decrease in numbers of CD62E+ MPs was observed 1 h after exercise for both patient and control samples. This decrease was present in both groups, but it was more pronounced in the patients.

Conclusions

Acoustic trapping technique may be used for isolation of MPs, including EMPs, from human plasma. The presented results indicate an anti‐inflammatory effect of physical exercise on the endothelium.

Support or Funding Information

Swedish foundation for strategic research
Cellular and molecular components regulating social behaviour of male Drosophila
Authors: Kim Woo Jae , Jan Lily Yeh , Jan Yuh Nung
Published: November 2023

Environmental conditions, such as the socio‐sexual context, can change rapidly. In these circumstances, the ability of animal of a genotype to express a different phenotype according to the environment can improve survival and reproductive success. A primary function of males for many species involves mating with females for reproduction. For males, current mating investment can often be subject to trade‐offs with future mating opportunities to maximize fitness in a rapidly changing socio‐sexual environment. Males have a finite resource for reproduction; hence a ‘time investment strategy’ has evolved to maximize reproductive success. Drosophila males respond to the presence of rivals by prolonging mating duration to increase the chance of passing on genes. In previous studies, we examined the genetic network and neural circuits that regulate rival‐induced longer mating duration (LMD). LMD can be induced solely via visual stimuli. LMD depends on the circadian clock genes timeless and period, but not Clock or cycle. LMD involves the memory circuit of the ellipsoid body (EB). Further, we identified a small subset of clock neurons in the male brain that regulate LMD via neuropeptide signaling.

More recently, we investigated a novel behavioral plasticity whereby male fruit flies exhibit a shortened mating duration when sexually satiated, called ‘Shorter‐Mating‐Duration (SMD)’. Both sexual experiences and contact‐based chemoreception is necessary to induce SMD. SMD requires the sexually dimorphic Gr5a‐positive neurons to detect female body pheromones, and can be induced by gustatory stimuli. The memory circuitry within the ellipsoid body (EB) and mushroom body (MB) brain regions is crucial to process this satiety state. SMD depends on the circadian clock genes Clock and cycle, but not timeless or period. SMD also relies on signaling via the neuropeptide sNPF, but not PDF or NPF. Sexual experience modifies the neuronal activity of a subset of sNPF‐positive neurons involved in neuropeptide signaling, which modulates SMD. Thus, our study delineates the molecular and cellular basis for SMD – a plastic social behavior that can be modified by sexual experience.

Our studies delineate part of the molecular and cellular basis of plastic behaviors elicited by different socio‐sexual environments. LMD together with SMD can be a model to study how the socio‐sexual environment affects on male behavioral plasticity via neural circuitry.

Support or Funding Information

HHMI

NIH R01

University of Ottawa Startup grant
Postprandial Hypertriglyceridemia‐induced Brain Vascular Inflammation by Next‐Generation Sequencing
Authors: Nyunt Tun , Wilson Dennis W. , Rutledge John C. , Aung Hnin H.
Published: November 2023

Vascular dementia (VaD), the second most common form of dementia, is associated with vascular risk factors, such as hyperlipidemia. The pathophysiological mechanisms and potential therapies for VaD are poorly understood; however, previous studies suggest a relationship of blood lipids and/or lipoproteins to vascular inflammation leading to dementia.

The objective of this study is to characterize brain endothelial cell responses to postprandial triglyceride‐rich lipoproteins (TGRL) and determine the pathways that have potential to increase risk for vascular dementia.

To address this question, we treated human brain microvascular endothelial cells (HBMVEC) with TGRL lipolysis products and then we used next‐generation sequencing (NGS)‐based RNA sequencing (RNA‐Seq) methods for transcriptional profiling experiments. Genome‐wide analyses of transcription of single nucleotides were generated on the Illumina HiSeq3000 sequencer. We then identified regulatory genes and their targets in RNA‐Seq datasets that have been differentially expressed using TopHat mapper and EdgeR statistical software packages. The Grinn software tool was used to retrieve pathway annotations of significant genes. The R package Piano was used for pathway enrichment analysis.

System biology analysis showed that TGRL lipolysis products activated 108 signaling pathways of which 48 signaling pathways were significantly activated. Among them, the most significant (p value=0) activated signaling pathways were MAPK (20 up‐regulated and 4 down‐regulated genes), TNF (16 up‐regulated and 4 down‐regulated genes) and Influenza A (10 up‐regulated and 4 down‐regulated genes). Stress response related genes (activating transcription factor 3 (ATF3), ATF4, Jun dimerization protein 2, sestrin 2, growth arrest and DNA damage inducible alph, DNA damage inducible transcript 3 (DDIT3) and DDIT4) and inflammatory response related genes (CCAAT/enhancer binding protein beta (CEBPB), VEGFA, gamma‐glutamylcyclotransferase 1, and TNF superfamily member 18, IL‐1 alpha) were up‐regulated and confirmed by qRT‐PCR analysis.

System biology analysis has led to the validation of putative lipid‐related targets and the discovery of 510 genes and corresponding isoforms that may be implicated in hypertriglyceridemia‐induced vascular inflammation in brain. Furthermore, the results of RNA‐Seq determined specific isoforms of key transcriptional regulator ATF3 and stress response signaling pathway in lipolysis product‐mediated brain vascular inflammation which may lead to vascular dementia.

Support or Funding Information

NIH‐NIA AG045541, NIH‐NIA AG039094, Richard A. and Nora Eccles Harrison Endowed Chair in Diabetes Research.
A systemic approach for evaluating biological function of acetyllysine in Danio rerio
Authors: Kim Sun Joo , Kwon Oh Kwang , Sung Eun Ji , Lee Sangkyu
Published: November 2023

Zebrafish (Danio rerio) is a widely used vertebrate model in genetics and cell biology. Although studies in several mammalian species have been performed, the mechanism of lysine acetylation in D. rerio embryos is incompletely understood. Lysine acetylation is an important post‐translational modification. Since the development of mass spectrometry (MS)‐based proteomics technology, important roles of lysine acetylation beyond histones have focused on chromatin remodeling during the cell cycle and regulation of nuclear transport, metabolism, and translation. Here, we investigated the global acetylome in D. rerio embryos by using an MS‐based system. We identified 351 acetylated peptides and 377 non‐redundant acetylation sites on 189 lysine‐acetylated proteins in 5‐day post‐fertilization (hpf) embryos of D. rerio. Among lysine‐acetylated peptides, 40.2% indicated 3 motifs: (ac)KxxxK, (ac)KxxxxK, and Lx(ac)K. Of 190 acetylated proteins, 81 (42.6%) were mainly distributed in the cytoplasm. Gene ontology enrichment and Kyoto Encyclopedia of Genes and Genomes pathway analyses showed that lysine acetylation in D. rerio was enriched in metabolic pathways. Additionally, 17 of 30 acetylated ribosomal proteins were evolutionarily conserved between zebrafish and humans. Our results indicate that acetyllysine might have regulatory effects on ribosomal proteins involved in protein biosynthesis.

Support or Funding Information

This work was supported by a National Research Foundation (NRF) grant funded by the Korean government (MSIP) (NRF‐2012R1A4A1028835 and ‐2014R1A1A2A16053716).
Optimized expression and purification of recombinant L‐asparaginase II: Tetramer stabilization by site‐specific covalent crosslinking of its subunits
Authors: Ramirez‐Paz Josell , Saxena Manoj , Griebenow Kai
Published: November 2023

L‐Asparaginase II is a tetrameric periplasmic enzyme from E. coli, which plays a major role in the chemotherapy of acute lymphoblastic leukemia due to its great affinity (26 μM) toward asparagine catalyzing its hydrolysis. Proteolytic degradation is one of the main causes of its inactivation.1,2 Our purpose is to study how the covalent cross‐linking of asparaginase's subunits can affect its activity, stability and proteolytic susceptibility. To accomplish this, specific amino acids on the monomer's surface will be mutated to cysteine. The subunits can then be covalently linked through thioether chemistry using different lengths of maleimide‐functionalized polymers. So far, we have been able to produced recombinant L‐asparaginase II in high purity (>99% by SDS‐PAGE) and yield (15–20 mg/L & 50% recovery). We optimized the expression and purification of our system, to come out with a straightforward method consisting of fast‐concentration → desalting → anion exchange, which allows rapid production of the recombinant asparaginase (only 5 hours). Mass spectroscopy was employed to confirm the identity of the recombinant enzyme in mature form (without N‐terminal peptide). In addition, circular dichroism and fluorescence spectroscopy suggest an identical secondary and tertiary structure when compared to the natural asparaginase. The presence of the enzyme tetramer and monomer was demonstrated by native‐PAGE electrophoresis. Finally, the specific activity of the recombinant asparaginase was found to be equal to the natural enzyme, compared by two different methods.3,4 At this point, we tested the expression of the mutant (A38C‐T263C) that will allow for the specific linkage, and confirmed that it is secreted as active enzyme. Now we will proceed with the cross‐linking of the subunits and evaluate activity, stability and proteolytic susceptibility of the neo‐conjugate vs. the non‐conjugate asparaginase.

Support or Funding Information

(1) Research facilities at the UPR ‐ Chemistry Department, Facundo Bueso

(2) Research facilities at the UPR‐Molecular Sciences Research Building

3) RISE Program at the University of Puerto Rico ‐ Rio Piedras Campus
Whole Cell Biosensor for Screening Lignin‐degrading Enzyme Library
Authors: Sana Barindra , Ramalingam Balamurugan , Raghavan Sarada , Seayad Jayasree , Ghadessy Farid John
Published: November 2023

Lignin is a potential renewable raw material for synthesis of various value‐added chemicals that can substitute fossil‐derived consumer products. Huge lignin biomass is produced as by‐product of paper industry while cellulosic components of plant biomass are utilized in paper pulp. Instead of vast potential, lignin remains the least exploited component of plant biomass due to its extremely complex cross‐linked three dimensional structures that cannot be efficiently degraded by currently available enzymes. Effective lignin degrading enzymes could be developed by enzyme engineering. Directed evolution is a strong tool for enhancing activity of currently available enzymes but application of this technique for improving efficiency of lignin degrading enzyme is limited due to unavailability of any high throughput screening method. With an objective of detecting the lignin degradation products (LDPs), we identified two promoters in E.coli genome that are up‐regulated by potential lignin degradation products. The promoters were identified by RNA‐Seq analysis of E. coli cells pre‐exposed to the sub‐lethal dose of a LDP for different exposure times. A ‘very green fluorescence protein’ (vGFP) gene was recombinantly placed under control of these promoters within a customized plasmid. The whole cell sensor was developed by transforming E. coli BL21 cells with these constructs. Although the constructs showed some leaky expression in the E. coli BL21 cells, the cells responded by increasing fluorescence while grown in the presence of the lignin degradation products (e.g. vanillin, acetovanillone and guaiacol), which was detected by fluorescence‐activated cell sorting (FACS) analysis. The sensors did not showed any increase of fluorescence by the presence of lignin or lignin model compounds. The fluorescence change by the presence of LDPs was dose‐dependent; one sensor can detect vanillin at the concentration as low as 0.5 mM. Both sensors showed best response to vanillin as an individual compound but better response was achieved with the mixture of the lignin degradation products.
Regulation of wago‐1: An Examination of the Regulatory Architecture that Governs Argonaute Protein Expression in Caenorhabditis elegans
Authors: Rosenberg Erin A , Youngman Elaine M
Published: November 2023

Across eukaryotes, thousands of small noncoding RNA molecules, in complex with Argonaute family proteins, silence or license gene expression through RNA interference (RNAi) and related processes. Since its discovery in 1998, RNAi has emerged as an invaluable tool for functional genomics, a critical player in regulation of endogenous gene expression, and a promising route to new therapeutics. Despite the important role RNAi plays in the regulation of gene expression, little is known about how the Argonaute effector proteins involved are themselves regulated. As a foray into understanding the regulation of the Argonaute protein network, we have chosen to examine the developing germ line of the nematode Caenorhabditis elegans, where at least five Argonautes are expressed in overlapping patterns. Presumably because the genome is being prepared for passage to the next generation and is relatively inaccessible to transcription factors, gene expression in the germ lineages of many animals is thought to rely heavily on posttranscriptional regulation. In C. elegans, a systematic study of 30 genes whose expression is known to be regulated in the germ line revealed that regulated gene expression in developing oocytes relied almost exclusively on sequences in the 3′ untranslated region (UTR), while sperm‐specific expression required promoter sequences but was largely independent of 3′ UTRs [2]. We therefore hypothesize that proper patterns of Argonaute expression in the germ line will depend both on promoter sequences and the 3′UTR. Using molecular cloning methods combined with CRISPR/Cas9‐mediated genome engineering based, we are using a GFP reporter transgene strategy to test this hypothesis. Thus far, I have generated a series of four reporter constructs that will allow me to individually assess contributions of the promoter and 3′UTR to regulation of the Argonaute protein WAGO‐1, which is expressed both in developing sperm and developing oocytes. I have designed and constructed CRISPR sgRNA sequences to guide integration of these transgenes into a defined locus on Chromosome II known to support germline expression, and am currently performing microinjection to establish transgenic worm strains. This research will provide the first insight into the mechanisms that regulate Argonaute protein expression during germ cell development, and will also advance methods for generating CRISPR‐integrated reporter transgenes in C. elegans.
Targeting the Young Scientist Community through their Interest in Biochemistry and Molecular Biology
Authors: Oquendo Mabel Giselle Pérez , Lachapel Ivana Serrano , Soto Luis Haddock , Lugo Juan Ramírez
Published: November 2023

The American Society for Biochemistry and Molecular Biology (ASBMB) Student Chapter at the University of Puerto Rico, Río Piedras (UPRRP), has as its primary focus to promote scientific interest in our students and the community. Given the growing need of researchers working on the many issues to be resolved in the field of Biochemistry and Molecular Biology, we want to genuinely inspire more people to get involved in this process to create and share knowledge. To inform about medical, technological and scientific advances, and encourage curiosity, and critical thinking, we performed journal clubs. However, to promote interaction between elementary, middle and high school students with university students who have scientific research experience, we accomplished community volunteer activities and outreach. As a result of our efforts in broadening scientific activities, our UPRRP chapter was awarded with ASBMB funds directed towards the organization of the “2nd Cellular and Molecular Biology Undergraduate Research meeting”. Through this symposium we achieved the participation of professors, graduate students, postdocs, and undergraduate students presenting local and summer research projects. Therefore, this meeting has been a platform for students interested in science, particularly in biochemistry and molecular biology area, are integrated into the scientific community. Conclusively, we aspire to build upon the foundations of the American Society for Biochemistry and Molecular Biology by serving as catalysts to inspire incoming students and everyone we may reach to contribute in our common pursuit of knowledge for the benefit of all.

Support or Funding Information

Regional Meeting Award ‐ ASBMB
The Vitamin D Content of a High‐Fat Diet Modulates Brain Reward Neurochemistry
Authors: Kubant Ruslan , Chatterjee Diptendu , Pannia Emanuela , Anderson G. Harvey
Published: November 2023

Background

Obesity is associated with increased consumption and preference for palatable foods as well as reduced blood levels of 25[OH]D3, which has been linked to altered dopaminergic and serotonergic signalling in the brain. The effect of vitamin D on high fat diet induced changes in brain reward systems is unknown.

Objective

To determine if vitamin D plays a role in modulating hedonic regulation of food intake in the brain in response to chronic consumption of a high fat diet.

Methods and Design

Male Wistar rats at 21 days of age were fed ad libitum for 14 weeks a high‐fat diet (60 kcal% fat) containing either: 1) the recommended amount (1,000 IU/kg diet) of vitamin D3 (HF‐RD) or 2) high vitamin D3 (HF‐HD, 7‐fold increase of the recommended amount). Rats fed a normal‐fat diet with the recommended amount of vitamin D3 (NF‐RD, 16 kcal% from fat) served as control. Body weight, food intake, serum 25[OH]D3, and concentrations of dopamine (DA), 3,4‐dihydroxyphenylacetic acid (DOPAC), serotonin (5‐HT), 5‐hydroxyindoleacetic acid (5‐HIAA) were measured in the nucleus accumbens (NAc) and β‐endorphin in the ventral tegmental area (VTA). DA and 5‐HT turnover rates were calculated by the ratio of their primary metabolites DOPAC to DA and 5‐HIAA to 5‐HT respectively, and used as an index of dopamine/serotonin transmission.

Results

The HF‐RD group consumed more calories and gained more weight than the NF‐RD group as well as exhibited increased DA (48%, P<0.05), DOPAC (330%, P<0.05), dopamine turnover (195%, P<0.05), 5‐HT (85%, P<0.05), 5‐HIAA (51%, P<0.05), and lower serotonin turnover (16%, P<0.05) in NAc. VTA levels of β‐endorphin were lower (46%, P<0.05) in the HF‐RD group compared to the NF‐RD. Supplementation of a HF diet with high vitamin D3 for 14 weeks resulted in a 3‐fold increase (P<0.05) in serum 25[OH]D3 and reduction (≈30%, P<0.05) in DOPAC levels and dopamine turnover compared to the HF‐RD group, but had no statistically significant effect on food intake, body weight, 5‐HT, 5‐HIAA, serotonin turnover rate or β‐endorphin levels in the brain.

Conclusion

The vitamin D content of a high‐fat diet modulates brain reward neurochemistry and may be a factor in neurotransmitter‐mediated regulation of the reward‐seeking behaviors.

Support or Funding Information

Canadian Institute of Health Research, Institute of Nutrition, Metabolism and Diabetes (CIHR‐INMD), Reference MOP‐130286.
Effect of caloric restriction on learning impairment in the hippocampus of ob/ob mice; through enhanced O‐GlcNAcylation and Ca2+ homeostasis
Authors: Lee Jong Youl , Jeon Byeong Tak , Heo Rok Won , Jeong Eun Ae , Yi Chin‐ok , Roh Gu Seob
Published: November 2023

Diabetes may adversely affect cognitive function, although the specific underlying mechanisms are unknown. On the other hand, caloric restriction (CR) increases longevity and improves memory. We examined the effects of CR on serum metabolic parameters and hippocampal protein expression in ob/ob mice, a model of obesity‐induced diabetes. We found that CR reduced hepatic steatosis and insulin resistance in ob/ob mice. In the hippocampus of these mice, CR increased the expression of O‐linked‐N‐acetylglucosamine (O‐GlcNAc) and GlcNAc transferase and decreased the expression of calcium/calmodulin‐dependent protein kinase P. It also decreased hippocampal lipocalin‐2 and phosphorylated tau. Furthermore, CR lessened the learning deficits that are typically seen in ob/ob mice. These findings indicate that CR may reverse obesity‐related brain glucose impairment and intracellular Ca2+ dysfunction and relieve learning impairment in diabetes.
Supplementation of Viscum album L. (mistletoe) might have a favorable effect on aging‐related muscle loss: A randomized, double‐blind, placebo‐controlled study
Authors: Lim Nam Ju , Kim Joohee , Kim Ji Yeon , Kwon Oran
Published: November 2023

Aging‐related muscle weak is associated with degenerative loss of skeletal muscle mass and function. The present study was investigated to test whether Viscum album L. var. coloratum Ohwi (Mistletoe) supplement can strengthen muscle mass and muscle strength in elderly subjects. A 12‐week, double‐blinded, randomized, parallel‐group, placebo‐controlled trial was performed in 63 subjects over 60 years of age (60–75y). Subjects were divided into three groups (n=21 for each group): placebo, low dose of mistletoe (1g/d) and high dose of mistletoe (2g/d). We evaluated muscle mass and function between the three groups over 12 weeks by measuring muscle mass [Body composition analysis (Inbody), and the cross‐sectional muscular area], muscle strength [Electromyogram(EMG), knee flexion‐extensor strength (Cybex) and grip strength], and short physical performance battery(SPPB) [usual gait speed, timed get‐up‐and‐go test and posturography]. The results showed that the percent of relative change in muscle mass was not significantly different between the groups after the intervention period. In muscle strength, the peak torque (Flexor, left, 120°) and the set total work (Flexor, left, 120°) of cybex increased significantly in high dose of mistletoe (P=0.017, P=0.002, respectively) compared to placebo group. In SPPB, the low dose of mistletoe was higher than other groups in changes of the 6‐minute gait speed (P=0.048). In conclusion, mistletoe was proven to beneficial role in muscle strength and function in older adults, regardless of its dose. In further study, the elucidation of molecular pathway which is associated with regulation of muscle biogenesis is needed to perform in biopsy muscle.

Support or Funding Information

This work was supported by the Ministry of Science, ICT & Future Planning (Bio‐synergy Research Project NRF2012M3A9C4048761) and BK21PLUS through the National Research Foundation.
Telifairiaoccidentalis (Pulp And Juice) Ameliorates High Sucrose Diet‐Induced Hepatic Steatosis And Oxidative Stress
Authors: Salau Bamidele Adewale , Odufuwa Kuburat Temitope , Adeniji P. O. , Atunnise Adeleke K.
Published: November 2023

Global increase in sucrose rich diet and beverages consumption has been linked with the recent escalation in fatty liver diseases and other oxidative stress. Also, there are numerous claims on the health benefits of vegetable rich diets, especially on cardiovascular diseases and other metabolic syndromes. Thus, this study investigated the ameliorative potential of Telifairia occidentalis juice and pulp portions on high sucrose diet‐induced hepatic steatosis, oxidative stress, electrolytes and hepatic biomarker enzymes status. Thirty two male albino rats of wistar strain weighing between 80 – 100g were used, grouped into six with respect to their dietary formulation. Sucrose supplied 30% energy portion of the high sucrose diet while Telifairia occidentalis supplied 10% energy of the diet supplemented with the leafy vegetable. The rats were fed with these modified diets for four weeks afterwards, the rats were sacrificed and liver was harvested for biochemical assay. In the results, high sucrose diet significantly (p<0.05) increased hepatic total cholesterol, triglycerides and LDL‐cholesterol, and reduced HDL‐cholesterol while, the vegetable fractions (pulp and juice) significantly (p<0.05) reduced the elevated lipid profile. High sucrose diet induced oxidative stress by reducing superoxide dismutase and catalase activities and resulted in oxidative damage as it significantly (P<0.05) increased malondialdehyde generation. Juice and pulp fractions of Telifairia occidentalis ameliorated the effect of high diet induced oxidative stress. Furthermore, high sucrose diet elevated alanine and aspartate transferases activities in the liver of rats. On the other hand, juice and pulp supplemented diets normalized the liver function biomarker enzymes activities. Although, pulp and juiced fractions of Telifairia occidentalis ameliorated the damaging effect of high sucrose diet on liver calcium, sodium and potassium status but not chlorides. Therefore, pulp and juiced forms of Telifairia occidentalis possess hypolipidemic, antioxidant, as well as hepato‐protective potentials also; pulp fraction of the Telifairia occidentalis tends to be more effective.
Vitamin C from Foods, but Not Supplements, Is Associated with Increased Cognitive Function in Older Adults
Authors: Davis Kristina , Chai Sheau C , Papas Mia A , Saltzgiver Julia , Falk Danielle , Caulfield Alexandra
Published: November 2023

As adults age there is an increased risk for developing dementia or cognitive impairment. Evidence suggests that there may be a relationship between a healthful diet high in antioxidants and preservation of cognitive function. Common antioxidants obtained from the diet include vitamins C and E, beta‐carotene, and selenium. It is unclear as to whether this relationship exists with antioxidants from foods, supplements, or total intake from foods and supplements. Therefore, the purpose of the present study is to assess average intake of vitamins C and E, beta‐carotene and selenium, from foods, supplements and total intake from foods and supplements, and the relation to Montreal Cognitive Assessment score, total Digit Span score, and Digit Span scaled score. 128 men and women between the ages of 65–80 were involved in a cross‐sectional study. Dietary information was obtained through a Block food frequency questionnaire and cognitive tests were administered by a trained researcher. Bivariate Pearson correlations were run to assess associations among variables. There were no significant associations between intake of any form of beta‐carotene, vitamin E or selenium. Vitamin C intake from foods was positively associated with both total Digit Span score (r=.231; p=.009) and Digit Span scaled score (r=.217; p=.014). In conclusion, findings suggest vitamin C intake from foods, but not supplements or total intake, is associated with increased cognitive function. Further research is needed to investigate the possible protective effects of dietary vitamin C intake in cognitive function.
24‐week exposure to dietary oxidized tyrosine (O‐Tyr) induces renal injury in rats through activation of p38 signaling
Authors: Li Zhuqing , Shi Yonghui , Le Guowei , Ding Yinyi , Zhao Qi , Tang Xue
Published: November 2023

Oxidized tyrosine (O‐Tyr) products such as Dityrosine (Dityr) and 3‐nitrotyrosine (3‐NT) have been widely detected in many commercial protein products. Since dietary O‐Tyr has been demonstrated to induce health harmful effects in our previous study, the objective of the present study was to examine renal toxicity of dietary O‐Tyr, focusing on oxidative stress and fibrosis changes. Male Sprague‐Dawley (SD) rats were treated with 0, 2, 4 and 8 g O‐Tyr/kg diet each day for 24 weeks. A group of rats treated with 8 g Tyr/kg diet were set as Tyr control. The kidney index, urine volume (UV), serum creatinine (Scr), blood urea nitrogen (BUN) and oxidative stress indexes were tested to evaluate renal injury. Expression of extra cellular matrix (ECM) contents, including hyaluronic acid (HA), laminin (LN), procollagen type Icarboxy terminal peptide (ICTP) and type I procollagen peptide (PINP), together with Masson staining of kidney slices was tested to investigate renal fibrosis. mRNA and protein expression related to p38 pathway was detected using the method of quantitative real‐time reverse transcription PCR, western blot and immunohistochemical staining. Our results showed that 24‐week feeding of O‐Tyr elevated kidney index, UV, Scr and BUN levels. Renal oxidative stress was evidenced by increased levels of ROS, protein carbonyl (PC), Dityr, 3‐NT, advanced oxidative protein products (AOPPs), malondialdehyde (MDA) and decreased activities of glutathione peroxidase (GPX), superoxide dismutase (SOD) and catalase (CAT). The expression of transforming growth factor‐β (TGF‐β), tumor necrosis factor‐α (TNF‐α), interleukin‐1β (IL‐1β) and IL‐6 in the kidneys increased after O‐Tyr treatment, suggesting renal inflammation. O‐Tyr also distinctly increased the phosphorylation of p38, enhanced fibrosis‐related TGF‐β1 and Smad 2/3 expression. In addition, elevated ECM contents, including HA, LN, ICTP and PINP, accompanied with results of Masson staining also supported renal fibrosis changes. All these changes are dose‐dependent in O‐Tyr treated group. Overall, our results suggested that O‐Tyr induces renal oxidative stress and renal fibrosis, which may be mediated through the activation of p38 signaling.

Support or Funding Information

The present study was supported by the 12th Five‐Year Plan for for Science and Technology Development (No. 2012BAD33B05)
Food Allergy Awareness and Nutritional Management by Preschoolers’ Parents and Faculties of Childcare Centers
Authors: Kim Junghee , Kim Soobin
Published: November 2023

This study was done to investigate the awareness and nutritional management of preschoolers with food allergy by their parents and faculties of childcare centers. Total 158 parents of preschoolers and 171 faculties of 137 childcare centers were recruited between July and September, 2015. The questionnaire for parents included a modified International Study of Asthma and Allergies in Childhood(ISAAC) questionnaire and questions regarding food frequency and knowledge related to food allergy. The preschoolers were divided into 3 groups, food allergy(FA), other allergy(OA), and non allergy(NO) groups. The rate of preschoolers who had experienced food allergy was 38.6% and diagnosed as food allergy by doctor was 17.7%. Family history of allergy significantly (p<0.001) affected allergic group. Forty nine preschoolers(80.3%) with food allergy had food restriction and twenty three preschoolers(37.7%) had self‐restriction without diagnosis. The major allergenic foods were eggs, milk, wheat, processed foods in FA group. The consumption frequency of allergenic foods in FA group such as ramyeon, noodles, bread, eggs, yourt and ice cream were significantly lower than those of the other two groups. Parents’ overall food allergy‐related knowledge was insufficient. Only 16.5% of parents had received education about food allergies. Most of parents wanted substitute menu for children with food allergy. Analysis on food allergy management by childcare centers showed that 70% of childcare centers asked parents about their children's allergic disease and 151 children from 67 childcare centers had food allergies. But 21% of childcare centers didn't know food restriction for food allergy children. Only 14% of childcare centers supplied substitute foods with the same amounts and types of nutrients. Only 28% of faculties received training about food allergies. Most of faculties wanted food allergy‐related supports. Overall results showed a lack of food allergy training for preschooler's parents and faculties of childcare centers. Therefore it is necessary for both parents and childcare center faculties to get comprehensive training on food allergy for better management. Particularly, food allergy‐related supports such as substitute menus without allergenic ingredients, guidelines for food allergy and anaphylaxis emergency care, etc, should be provided for parents and faculties