bluuebunny/crossref_metadata_2025_split huggingface dataset:
Unique DNA Polymerase kappa Interactome Suggests Novel Cellular Functions
Authors: Paul Shilpi , Cifani Paolo , Pappin Darryl , Spratt Thomas 
Published: May 2022

Translesion DNA synthesis (TLS) polymerases evolved to tolerate DNA damage that bypasses DNA lesions, thus preventing genomic instability. Multiple TLS polymerases exist with different damage tolerance capabilities, since they have low fidelity their access to the replication fork must be regulated to minimize mutations. The current paradigm is that a combination of kinetic partitioning and protein‐protein interactions are used to regulate TLS polymerase activity. A major knowledge‐gap in elucidating the roles of these polymerases is that it is difficult to identify which polymerase is active in a specific situation. We designed and synthesized a novel nucleotide analog N2‐benzyl‐ 2′‐deoxyguanosine (EBndG) that is highly selective toward DNA polymerase kappa (Pol κ), a Y family TLS polymerase. Pol k can bypass bulky lesions in the minor groove generated by a metabolite benzo[a]pyrene diolepoxide (BPDE), an environmental carcinogen. Although Pol κ has been identified to have multiple cellular roles, the mechanisms regulating its different cellular activities are unknown.To interrogate the identity of proteins surrounding the Pol κ active sites, we performed an extensive study using modified iPOND (isolation of proteins on nascent DNA), called iPoKD (isolation of proteins on Pol kappa synthesized DNA). Human cell lines were treated with BPDE, subsequently 5‐ethynyl‐2′‐deoxyuridine (EdU) or EBndG was added and proteins bound to the DNA containing EdU and EBndG were analyzed by mass spectrometry. In addition, we performed quantitative analysis of the Pol κ active sites interactome using iPoKD followed by iTRAQ (isobaric tags for relative and absolute quantitation). Our data identified DNA replication and repair proteins previously identified with EdU pull‐downs; and interestingly enrichment of RNA binding, ribosome biogenesis, nucleolar proteins and transcriptional repressive complex(es) associated with EBndG pull‐downs. Chromatin modifiers, histone chaperones and histone variants are identified suggesting changes in chromatin structure that facilitates Pol κ‐mediated DNA lesion bypass, repair and restorative process. identification of unique proteins associated with EBndG‐containing DNA, suggests novel roles for Pol κ’s activity in the cell.Using super‐resolution confocal microscopy, Pol κ activity is identified in the nucleolus after BPDE damage. EBndG is observed in the nucleolar DNA and Pol κ 's activity regulated by the canonical polycomb‐complex recruited by the PARylation of PARP1. BPDE lead to transcriptional stress and repression that is gradually recovered. We are investigating whether Pol κ maintains ribosomal DNA integrity after BPDE damage and is required for TLS DNA synthesis or repair. Pol κ active site associated candidate proteins are being validated using CRISPR‐Cas9 knockout or siRNA knockdown strategies. These data will provide first insight into Pol κ’s core interactome, it's regulation, chromatin surrounding Pol κ active sites and its novel cellular roles.
Hepatoprotective effect of a Novel Functional Drink based on Ecuadorian Plant Species
Authors: Paladines‐Santacruz Geovanna L. , Orellana‐Manzano Andrea , Manzano Patricia , Zaruma‐Torres Fausto , Ortíz‐Ulloa Johana , Cárdenas Washington , Torres‐Arias Marbel , Seqqat Rachid , Vanden Berghe Wim 
Published: May 2022

Liver diseases are considered a severe public health problem involving disorders that alter liver function. Since the liver plays an essential role in the metabolism and detoxification of substances that enter the organism, it is susceptible to damage caused by xenobiotic agents such as viruses, alcohol, and drugs. Acetaminophen is an over‐the‐counter drug widely used as an antipyretic and analgesic. It is safe at therapeutic doses, but it is easily overused, causing severe liver diseases. In a previous study, our investigation group developed a functional drink from the aqueous extracts of Ilex guayusa, Vernonanthura patens, and cocoa husks. This combination presented various bioactive compounds with antioxidant properties that could help reduce oxidative stress in pathologic conditions. For the above mentioned, the study's objective was to evaluate the hepatoprotective effect of the beverage through an in vivo test. Male mice were treated with two formulations (pre‐formulated and microencapsulated) of the functional drink in three concentrations for seven days as a preventive oral treatment. After that, mice were induced to liver damage at a single intraperitoneal dose of acetaminophen (250 mg/kg of body weight). The evaluation was based on the liver damage biomarkers, ALT, and AST; anatomopathological studies were also evaluated. The results showed that the best formulation as an oral pre‐treatment was the pre‐formulated. The pre‐formulated beverage attenuated the injury caused by the acetaminophen by inhibiting serum ALT and AST activities and reducing liver macro and microscopic liver alterations. The successful doses were at 250 and 1000 mg/kg b.w. The microencapsulated did not present adequate liver protection at any doses evaluated because it has no statistical difference with the toxic model group.In conclusion, the present study demonstrated for the first time that the functional drink effectively prevented acetaminophen‐induced liver injury in vivo. Our data indicated that the pre‐formulated was the most effective pre‐treatment to attenuate the hepatic injury. These results suggested that this beverage possessed antioxidative activity that likely contributed to its protective effect against acetaminophen liver damage.*Corresponding Author: Orellana‐Manzano Andrea, email: akorella@espol.edu.ec
The effect of novel second generation prebiotics (levan of β‐[2,6]‐glycosidic linkages) on the cardiovascular system in obesity
Authors: Sahyoun Amal , Kwon Young‐In , Munkhsaikhan Undral , Karboune Salwa , Kassan Modar 
Published: May 2022

ObjectivesBacteria dysbiosis is very well documented in obesity and is known to regulate the cardiovascular system. Recovering the bacteria composition is a key element in improving the cardiovascular function during obesity.HypothesisIn this study we investigate the effect of levan, a novel second generation of prebiotic (β‐[2,6]‐linkages, on the cardiovascular system in obesity.MethodsEight‐week‐old C57/b6 mice were fed with high fat diet (HFD) for 12 weeks in the presence and absence of levan (β‐[2,6]‐linkages). Treatment was administered by gavage (250 mg/kg) for the last 30 days of HFD feeding. Body weight, blood glucose, body composition and lipid profile were determined. Vascular function and endothelial function markers were studied in aorta and mesenteric resistance arteries (MRA).ResultsLevan (β‐[2,6]‐linkages) treatment reduced the body weight in obese mice. Blood glucose, % of fat mass, total cholesterol, and LDL levels were significantly reduced and the % of lean mass was significantly increased after Levan (β‐[2,6]‐linkages) treatment. Aortic and mesenteric arteries vascular response to sodium nitroprusside (SNP) was similar among groups. However, the vascular response to acetylcholine (Ach) was improved in the treated group.ConclusionsTreatment with Levan (β‐[2,6]‐linkages) attenuated the increase in body weight in obese mice and restored the lipid profile and the vascular function.
Proteomic analysis of urinary exosomes from patients with lithium‐induced nephrogenic diabetes insipidus
Authors: Grønkjær Camilla , Apolito Luciano D. , Rej Soham , Fenton Robert , Trepiccione Francesco , Christensen Birgitte M. 
Published: May 2022

Lithium (Li)‐induced Nephrogenic Diabetes Insipidus (NDI) is a large healthcare problem, which affect around 40% of patients with bipolar disorder undergoing Li treatment. NDI is characterized by the inability in the kidney to concentrate urine in response to vasopressin in the kidney collecting duct (CD). In animals, Li causes a downregulation of aquaporin‐2 and a cellular remodeling of the CD in rats resulting in a reduced fraction of principal cells relative to intercalated cells. Whether Li has similar effects in the human kidney is, however, not known. Urinary exosomes are extracellular nanovesicles derived from intracellular multivesicular bodies fused with the plasma membrane of epithelial cells lining the kidney tubular system and the urinary tract. The purpose of the first part of this study is to analyze whether the urinary rat proteome reflects the Li‐induced changes in the kidney. This is followed by proteomic analyses of urinary exosomes isolated from Li‐NDI patients.Rats were divided in three groups; controls (n=8), 2‐week Li treatment (40 mmol/kg/food, n=8) and 4‐week Li treatment (40 mmol/kg/food, n=8). Treatment with Li showed a significant increase in urine output after 2‐weeks (P<0.01) and 4‐weeks (P<0.05). Additionally, the rats developed severe polydipsia, with water intake significantly increased after 2‐weeks Li treatment (P<0.001) and 4‐weeks Li treatment (P<0.0001).Early morning urine samples from the placebo group in a previous published 12‐week placebo‐controlled pilot trial (Fotso Soh et al, 2021. A double‐blind, randomized, placebo‐controlled pilot trial of atorvastatin for nephrogenic diabetes insipidus in lithium users. Bipolar Disorders 23, 66–75) involving Li‐NDI patients (n=9) were collected. The Li‐NDI participants have been on a stable dose of Li (˃2 months) and determined to have a urinary osmolality <600 mOsm/kg at baseline. In addition, urine from random healthy controls (n=6) were collected. Urine samples from the rodent study and human study were subjected to ultracentrifugation for exosome isolation and will be further processed for mass spectrometry analysis (MS/MS).
Erythropoietin M2 macrophage Signaling Promotes Schwann Cells Clearance and Functional Recovery Following Peripheral Nerve Injury
Authors: Govindappa Prem Kumar , Elfar John C. 
Published: May 2022

BackgroundTraumatic peripheral nerve injury (TPNI) obliterates axons and Schwann cells (SCs) that must be cleared through phagocytosis to allow surviving SCs to de‐differentiate and divide and guide the regeneration process. In TPNI, macrophage (MØ) infiltration and early phenotypic transitions (M1 to M2) are critical for the phagocytosis of SCs debris. Delayed phagocytosis critically aggravates inflammation and impairs MØ function, which leads to failure to advance SC orchestration and neuronal regeneration. Erythropoietin (EPO), a pluripotent hormone, helps to guide MØs and SC transition post‐TPNI. The significance of the effects of EPO on debris clearance, apoptosis, myelination, and functional improvement is unknown, despite much effort motivating clinical translation of EPO for TPNI. We hypothesized that a role in supporting M2 phenotype macrophages after TPNI might explain EPO’s neuroprotective function through SCs debris clearance.Methods and ResultsWe evaluated EPO’s effect on sciatic nerve crush injury (SNCI) and found EPO treatment (5000 IU/kg, intraperitoneally, post‐surgery days 1, 2, and 3,) increased myelination and sciatic functional index (SFI) and augmented anti‐apoptosis and phagocytosis of myelin debris via CD206+ macrophages when compared to saline treatment in the mouse. EPO increased efferocytosis of apoptotic sciatic nerve derived Schwann cells (SNSCs) both in bone marrow derived macrophages (BMMØs) and peritoneal derived macrophages (PMØs) in‐vitro, as well as in PMØs in‐vivo by immunofluorescence (IF) and flow cytometry. EPO treatment significantly attenuated BMMØ pro‐inflammatory genes (IL1b, iNOS, and CD68) and augmented anti‐inflammatory genes (IL10, and CD163) in addition to the cell surface marker CD206 expression after lipopolysaccharide (LPS) induction. EPO also showed anti‐apoptotic (Annexin V/ 7AAD) effects in BMMØs. Our data demonstrate that EPO promotes the M2 phenotype macrophages to ameliorate SN apoptosis and efferocytosis of dying SCs and myelin debris and improves SN functional recovery following SNCI.ConclusionsTo the best of our knowledge, this is the first study to demonstrate EPO’s ameliorative effect on the progression of post‐TPNI via M2 phenotype macrophage phagocytosis of SCs debris with early anti‐apoptotic and anti‐inflammatory effects. These data may support the findings of other researchers and might shed light on the role of EPO in improving functional recovery in the traumatized limb.
Low output syndrome after cardiac surgery is mediated through extra corporal circulation induced RAGE activation
Authors: Velten Markus , Hilbert Tobias , Dewald Oliver , Meyer Rainer , Duerr Daniel 
Published: May 2022

ObjectiveCoronary artery bypass grafting (CABG) is a surgical procedure routinely performed using extracorporeal circulation (ECC). This on‐pump CABG is known to be associated with severe postoperative organ failure including cardiac dysfunction and low cardiac output syndrome. Mechanisms inducing cardiac dysfunction are complex but may include on‐pump CABG induced inflammation due to exposure of the blood to the bypass circuit, potentially increasing serum concentrations of inflammatory mediators. High‐mobility group box 1 (HMGB1) functions as an inflammatory mediator and has been shown to induce cardiovascular dysfunction in sepsis and impair cardiomyocyte contractility in experimental models. However, the impact of HMGB1 on on‐pump CABG induced myocardial dysfunction is not known. Therefore, we tested the hypothesis that on‐pump CABG associated increases in serum concentrations of inflammatory mediators induce cardiomyocyte dysfunction in vitro and investigated potential mechanism including the role of the HMGB1‐RAGE (receptor for advanced glycation end products) axis.ApproachAfter institutional revenue board approval, informed consent, and IACUC approval, serum samples (n = 18) were collected from CABG patients pre, 1, 6, and 24 h after aortic de‐clamping. TNFa, IL‐1b, IL‐6, HMGB1, and sRAGE serum concentrations were analyzed by ELISA. Primary murine cardiomyocytes were incubated with patient serum and single cell contractility was assessed at frequencies between 0.5 and 10Hz using an optical contractility system (IonOptics LLC, Westwood, MA).ResultsIL‐6, HMGB1, and sRAGE serum levels were increased after CABG (Figure 1). In preliminary studies we incubated murine cardiomyocytes with human serum collected from healthy volunteers and determined the highest serum concentrations that did not affect cardiomyocyte contractility. Cardiomyocytes were incubate with human serum and serum collected pre‐surgery had no effects on murine cardiomyocyte contractility. However, primary murine cardiomyocytes incubated with serum collected 1h and 6h after aortic de‐clamping showed a significant decrease in % peak shortening. Pre‐incubation with the RAGE antibody FPS‐ZM1 (Cayman, Hamburg, Germany) prevented the serum induced cardiomyocyte dysfunction (Figure 2).Conclusionon‐pump CABG increased protein serum concentrations of inflammatory cytokines including HMGB1. HMGB1 has been shown to cleave and activate RAGE. sRAGE serum levels have been increased after on‐pump CABG. Deactivating RAGE with the high‐affinity inhibitor FPS‐ZM1 prevented serum induced cardiomyocyte dysfunction, suggesting that the HMGB1‐RAGE axis is mediating on‐pump CABG associated low‐cardiac‐output syndrome.
Mnemonics Improve Pharmacy Students' Knowledge Retention, Clinical Application, and Exam Performance in a PharmD Pharmacology Course
Authors: Munusamy Shankar , Koenigsfeld Carrie , Torry Ronald 
Published: May 2022

BackgroundStudents in healthcare professions taking pharmacology courses often feel challenged with learning and remembering crucial facts about medications, i.e., knowledge acquisition and retention. In addition, applying pharmacological knowledge into clinical settings consumes significant amounts of the student’s working memory (i.e., cognitive load). Mnemonics (memory aids) are used to reduce cognitive load and facilitate learning and knowledge retention among students. Our study assessed the impact of the mnemonics' use in a Pharmacology course offered to second‐year pharmacy students on students' perceptions of knowledge acquisition, retention, and clinical application in correlation to their exam performance.MethodsEighteen mnemonics were developed and used in a course covering endocrine and autonomic pharmacology topics. Students' perceptions of the impact of mnemonics' use on their knowledge acquisition, retention, clinical application, critical thinking, and confidence while answering questions on exams were collected using an anonymous survey in the fall semester of 2020. Further, students' perceptions were matched to their exam performance.ResultsSurvey responses from 56 students (85% response rate) indicated that 80.4% agreed that mnemonics' use facilitated knowledge acquisition by reducing anxiety while learning, and 98.2% of students indicated that the use of mnemonics improved knowledge retention and application. Mnemonics' use was also reported to improve critical thinking while answering exam questions (89.3% agreed) and increased confidence during test‐taking (96.4% agreed). Interestingly, 64.7% of students using mnemonics answered the associated exam questions correctly compared to 35.3% of students (p<0.05) who did not use the mnemonic aids.ConclusionsOur study suggests that mnemonics' use in pharmacology courses not only improved students' perception of knowledge acquisition, retention, application, and critical thinking but also improved their exam performance on specific questions that tested concepts taught with mnemonics. Future studies will assess students' knowledge retention and perceptions of mnemonics' use with exam performance in subsequent pharmacotherapy and pharmacy skills courses.
Profiling Growth Patterns of Human Gut Microbes in Response to Preterm Human Milk and Formula
Authors: Engevik Mindy A. , Stripe Leah A. , Baatz John E. , Wagner Carol L. , Chetta Katherine E. 
Published: May 2022

BackgroundThe infant gut microbiota undergoes significant fluxes in microbial composition during the first few years of life. The intestine is colonized shortly after birth with facultative anaerobes, like as Enterococcus, Enterobacter, Streptococcus, and Staphylococcus, which reduce the intestinal oxygen content and create niches for strict anaerobes, like Bifidobacteria, Bacteroides, and Clostridia. The dominant factors which influence the infant gut composition are gestational age and diet. The gut microbiota of preterm infants is characterized by limited microbial diversity and increased levels of facultative anaerobes such as Enterococci, Staphylococci, and Enterbacteriacea (Enterobacter, Escherichia, and Klebsiellaspp.). The gut microbiota is also highly influenced by diet, specifically formula and human milk use. The intake of human milk is associated with increased Bifidobacteria, while the intake of formula correlates with increased Enterobacteriaceae. This study aimed to analyze next‐generation products including preterm formula, human milk‐oligosaccharide term formula, and preterm breastmilk using a culture‐based model.Methods & ResultsWe cultured 8 different Bifidobacteria, 8 Lactobacilli, 8 Streptococcus, 3 Enterococci and 4 Klebsiella strains in a fully defined bacteria media and monitored growth and pH after 20 hrs of anaerobic incubation. Significant differences were identified in strain growth and culture pH between human milk and formula. Human milk significantly elevated the growth of all Bifidobacteria and Lactobacilli. In contrast, human milk supported the growth of three Streptococci and only nominally supported pathobiont Enterococci and Klebsiella strains. Preterm formula elevated the majority of Bifidobacteria and Lactobacilli, as well as several Streptococci. Interestingly, preterm and full‐term formula significantly enhanced the growth of pathobiont strains. Full‐term formula only supported the growth certain commensal microbes, which was in marked contrast to human milk and preterm formula. Using computational modeling, a correlation was observed with human milk oligosaccharide (HMO) degrading glycosyl hydrolase profiles in the genomes of commensal microbes with human milk, but not with formula; confirming the HMO preference of our representative infant commensal microbes.ConclusionsThese findings indicate a unique profile of growth in response to human milk and formula. Since alterations in the infant gut microbiota have linked to childhood health and development, understanding how bacteria differentially utilize whole milk or formula may provide insights into which microbes drive diet‐related microbiota shifts.
The Magnitude of Hidden Hunger and Cognitive Deficits of Children Living in Some Selected Orphanages in Kumasi, Ghana During the COVID Pandemic
Authors: Asamoah Mary , Tandoh Marina 
Published: May 2022

Many studies have assessed the magnitude of mixed micronutrient deficiencies or individual micronutrient deficiencies among children under 5 years, women of reproductive age (15‐ 49 years old) and pregnant women. This has led to various interventions for these population groups including supplementations, fortifications etc. However, the same attention has not been given to vulnerable children living in various orphanages, especially in Children’s Homes in Ghana where much is not known about their nutritional status. Socio‐ economic downturns like that induced by the current coronavirus pandemic affects food security and nutrition, thus the nutritional status of this vulnerable population could potentially be worsened.ObjectiveThis study assessed the magnitude of hidden hunger and cognitive deficits of 130 children (6‐ 13 years old) living in three selected orphanages in Kumasi, Ghana.MethodologySociodemographic data, anthropometric assessment (BMI‐for‐age and height‐for‐age), dietary assessment (3‐day repeated 24‐ hour dietary recall), urinary iodine levels and cognitive performance (Raven’s Coloured Progressive Matrices) were assessed.FindingsBoys formed 50.8% of the study population while girls formed 49.2%. The median age of participants was 10.50 years. About 12.3%, 7% and 10.0% of participants were stunted, thin and overweight/ obese respectively. Prevalence of mild iodine deficiency (i.e. 50‐ 99 μg/L) was 16.2%. Iodine deficiency was significantly higher (16.2%) in participants who had lived for at least 7 years in the orphanage compared to those who had lived less than 7 years (23.6%) (p = 0.047). About 17% of participants performed poorly (< 50%) in the cognition test. Mean cognitive test scores were significantly different among the orphanages (p = 0.027). Majority of participants, 89.2%, 54.6%, 76.9% and 77.7% had adequate intakes for iron, zinc, vitamin C and folate respectively whereas intake of vitamins A and B12 were inadequate for majority of participants (90.8% and 50.8% respectively). There was no significant correlation between micronutrient intake and cognitive performance. However, mean cognition test scores were significantly different between participants who had adequate and inadequate intake of iron and vitamin A (p = 0.007 and p < 0.001 respectively).The significance of the research is to provide data for relevant policy makers and stakeholders involved in the care of orphans and vulnerable children in Ghana like the Social Welfare Department. It is also expected that this research will stimulate further studies in the occurrence of multiple micronutrient deficiencies in vulnerable children since nutrition is not only necessary for physical development, but also for cognitive development and life‐long health (WHO, 2016).ConclusionFindings of this study warrant a closer look at nutritional intakes in orphanages to improve on hidden hunger.