bluuebunny/crossref_metadata_2025 huggingface dataset:
The Effect of Magnolol on a Polycystic Ovary Syndrome Rat Model with Insulin Resistance
Authors: Kanaan Amjad , Khachab Maha , Nassar Aline , Zaatiti Hayat
Published: April 2020

Polycystic ovary syndrome (PCOS) is a prevalent reproductive and metabolic disorder. Insulin resistance (IR) is highly associated with and aggravates the symptoms of PCOS. The Insulin sensitizing agents, Thiazolidinediones, are PPARγ agonists that improve many of the symptoms of PCOS. Magnolol is a natural PPARγ agonist that improves insulin sensitivity in experimental models. Using a dehydroepiandrosterone (DHEA)‐induced rat model of PCOS and IR, we aim to explore both the potential beneficial effect and the molecular mechanisms of action of Magnolol. Post‐pubertal female Sprague Dawley rats, injected subcutaneously with DHEA (6mg/100g body weight) for 28 days, exhibited a significant increase in body weight (242.5 ± 5.4 grams) as compared to age‐ and weight‐matched controls (226.8 ± 4.7 grams). The DHEA‐injected rats developed an increased number of cystically dilated follicles with thicker granulosa as compared to control. PCOS rats showed statistically significant elevated serum levels of Luteinizing hormone (LH) (10.0 ± 2.7 ng/ml) and fasting insulin (2.18 ± 0.9 ng/ml); with an increased Homeostatic Model Assessment Index of insulin resistance (HOMA‐IR) (9.98 ± 4.9) as compared to controls (6.3 ± 0.2 ng/ml; 0.6 ± 0.3 ng/ml; 2.7 ± 3.9 respectively). Compared to control, PCOS rats had a significant lower ovarian protein expression of peroxisome proliferator activated receptor gamma (PPARγ), insulin receptor substrate 1 (IRS1), and protein kinase B (Akt) by Western Blot. Conversely, the PCOS group showed an increased mammalian target of rapamycin (mTOR) pathway activity as evident by an increase protein expression of phosphorylated mTOR as compared to control. When treated for 28 days with oral Magnolol (500mg/kg), the DHEA‐induced PCOS rats showed a statistically significant decrease in body weight (231.6 ± 13 grams) and serum LH levels (3.9 ± 1.8 ng/ml) as compared to the non‐treated PCOS rats (250.3 ± 9.7 grams; 9.5 ± 5.7 ng/ml respectively). The number of cystically dilated follicles in the Magnolol‐treated PCOS rats was significantly reduced compared to the non‐treated PCOS rats. In the Magnolol‐treated PCOS rats the ovarian protein expression of PPARγ, IRS1 and Akt was significantly increased, while the mTOR expression was decreased compared to the non‐treated PCOS group. In conclusion, Magnolol ameliorates the DHEA‐induced PCOS phenotype histologically, hormonally and metabolically. Fundamentally, this work explores the elusive pathophysiologic association between IR and PCOS, by targeting pathways common to both disorders.

Support or Funding Information

This work is supported by the University of Balamand, Faculty of Medicine and Medical Sciences
Ansa cervicalis and hypoglossal nerve topographical relation: anatomical study on cadavers.
Authors: Boog Gustavo Henrique Pereira , Medeiros Vitor Macedo Brito , Chinelatto Lucas Albuquerque , Hojaij Flávio Carneiro , Akamatsu Flávia Emi , Jácomo Alfredo Luiz
Published: April 2020

Ansa cervicalis and hypoglossal nerve are important neural structures of the neck. Their topographical disposition have important clinical and surgical implications. As described in a recent consensus, the ansa cervicalis originates in the spinal cord and then joins the hypoglossal awhile, without exchanging fibers, diverging from it to innervate the infrahyoid muscles. However, some dissection studies show individuals in which ansa cervicalis and hypoglossal nerve follow independent trajectories, diverging of classical characterization. This anatomical variation supports the need for better description of the link between these nerves and, therefore, improve surgical planning and technique. This project objective is to better characterize the relationship between ansa cervicalis and hypoglossal nerve, describing the percentage of link between these structures. In this regard, it will be realized a cross‐sectional study with 15 cadavers bilateral neck dissections. The study plans to analyze the data through nonparametric statistics. Our analysis will assess correlations between the topographic variety and sex, age, ethical and anthropometric characters of individuals. So far, we dissected 5 cadavers; of these 4 (80%) have ansa cervicalis not connected to hypoglossal nerve, meanwhile just 1 (20%) have this link. The project schedule predicts 10 more corpses dissections until January 2020. Until now, the small sample limit statistical analysis, but certainly it suggests there is a significant anatomical variant in the brazilian population. It is expected that our final results will contribute to better comprehension of these important neck structures and improve medical practice.
Cloning and expression of a novel α‐amylase from Salinispora arenicola CNP193 in E. coli
Authors: Ahmed Sibtain , Fang Yaowei
Published: April 2020

A novel gene encoding α‐amylase was cloned from marine bacterium Salinispora arenicola CNP193 and the protein was expressed in Escherichia coli. The α‐amylase gene from S. arenicola CNP193 had a length of 1839 bp and encoded a α‐amylase with an estimated molecular mass of 74 kDa. The optimum temperature and pH for the recombinant α‐amylase was 50 °C and 7 respectively. Na+, K+ and Ca2+ increased the activity of the recombinant α‐amylase whereas the enzyme was inhibited by Cu2+, Zn2+, Hg2+, Pb2+, Fe3+ and Mn2+. Thin layer chromatography results confirmed that monosaccharide, disaccharide and maltotriose are the hydrolysis products. The results of our study suggest that this enzyme has considerable potential in industrial applications.
Whitening Effect of Radiation‐resistant Paenisporosarcina quisquiliarum 17mud 1‐1541 Extracts
Authors: Park Yuna , Kim Myung Kyum
Published: April 2020

Paenisporosarcina quisquiliarum 17mud 1‐1541 (designated to KCTC13860BP), an extremophilic radiation‐resistant bacterium, was newly isolated from the mud in coastal areas of Korea. The aim of this study was to evaluate the whitening effects of 17mud 1‐1541 extracts. The viability of B16F10 cells was evaluated by MTT assay to determine the non‐toxic doses for further experiments. The contents of melanin and the cellular pigmentation images were evaluated. To confirm 17mud 1‐1541 mechanism in melanogenesis, cellular tyrosinase, L‐DOPA, and α‐glucosidase activities were determined. 17mud 1‐1541 has strong inhibitory effects on melanin production and cellular tyrosinase activity in α‐MSH‐induced B16F10 cells. Melanogenesis‐related factors such as L‐DOPA oxidation, tyrosinase and α‐glucosidase activities were all reduced in the presence of 17mud 1‐1541. Overall, 17mud 1‐1541 has a whitening effect due to inhibition of melanogenesis, which is affected by α‐glucosidase and tyrosinase inhibition activities. Our findings suggest that 17mud 1‐1541 has a promising potential to be used as skin lightening agent.
Investigating L1 promoter activity during mouse development using LacZ transgenes
Authors: Saha Partha S. , LaCanne Trenton , An Wenfeng
Published: April 2020

Long interspersed elements type 1 (L1s) are highly abundant retrotransposons in the human genome. To fully understand the role of L1s during development and disease, it is important to track L1 expression and epigenetic regulation at individual L1 loci throughout development at the cellular level. However, it is extremely technically challenging to monitor transcriptional activities of individual endogenous L1s due to high sequence homology. Toward this goal, we created independent single‐copy LacZ transgenic mouse lines, in which the LacZ reporter is regulated by a copy of the endogenous mouse L1 promoter. To control for the effect of gene‐body DNA methylation on L1 promoter activity, we also generated independent mouse lines carrying the CpG‐less version of the LacZ, termed LacG. After mapping of the transgene locus in each line, mouse tissues were stained with X‐gal to visualize L1 promoter activity at single cell level. QuPath was used to quantify X‐gal positive cells in whole‐slide scanned brain sections. We found that the LacG reporter supported robust expression as compared to the LacZ reporter. However, there were substantial variations among the LacG reporter lines, in which the transgene is integrated in random genomic loci. Intraline variations of expression in the same organ might be dictated by generation and age, which is yet to be established statistically. Orientation dependent regulation of L1 promoter was also studied by targeting the LacG transgene in a specific intronic locus in either sense or antisense orientation. Interestingly, antisense orientation showed augmented expression than its counterpart. Ongoing effort aims to reveal the pattern of these signals in different anatomical regions of brain and also in other major organs, which might support L1 retrotransposition in different developmental period in these mouse models. The goal is to provide a comprehensive mapping of L1 promoter activity in a cell dependent and locus dependent manner. Such information will be instrumental in our understanding of L1’s involvement in physiological and pathological conditions, including embryogenesis, gametogenesis, and under environmental stress.

Support or Funding Information

Supported by NIH R21HD080143 and R21OD017965.
Maternal FUT2 genotype in relation to risk of acute infections and rotavirus vaccine shedding in infancy
Authors: Adkins Grace E. , Thorman Alexander , Conrey Shannon C. , Cline Allison R. , Staat Mary A. , Payne Daniel C. , Piasecki Alexandra M. , Burke Rachel M. , Bowen Michael D. , Morrow Ardythe L.
Published: April 2020

The human FUT2 gene codes fucosyltransferase2, an enzyme that catalyzes the production and secretion of H‐antigen in bodily fluids and tissues. FUT2 status has been associated with risk of infection for some pathogens. In addition, mothers who are homozygous positive or heterozygous for the FUT2 gene produce a class of oligosaccharides in their milk that can act as decoy receptors for some selected pathogens. The link between maternal FUT2 status and illnesses in breast‐fed infants is not clear.

The Pediatric Respiratory & Enteric Virus Acquisition and Immunogenesis Longitudinal (PREVAIL) Cohort study of mother‐infant pairs in Cincinnati, OH assessed whether an epidemiologic association may exist between maternal FUT2 genotype and incidence of acute gastroenteritis (AGE), acute respiratory infection (ARI), and rotavirus vaccine shedding in their infants. We expected that the exclusively breastfed children of FUT2+ mothers would have lower incidences of infection due to the unique breastmilk oligosaccharide composition.

We included all 75 mother‐infant pairs who exclusively breastfed their infants, which provided a total of 1,243 child‐weeks of follow‐up from birth to 6 months. All mothers were genotyped for FUT2 by PCR. Of these, 40 (53%) infants were vaccinated against rotavirus and had ≥1 weekly stool sample collected within 8 weeks after vaccination and tested by real time RT‐PCR assays to determine vaccine shedding. Infant infection data came from weekly maternal reports of AGE and ARI that were collected by using an automated text messaging system.

Infants of FUT2+ mothers were found to have longer duration of rotavirus shedding post‐vaccination than babies of FUT2− mothers (p=0.007). Additionally, infants of FUT2+ mothers had more AGE symptomatic study weeks (5.6%) than children of FUT2− mothers, who had no symptomatic AGE weeks (p<0.001). Similarly, infants of FUT2+ mothers had ARI with fever for approximately 2.7% of study weeks, whereas babies of FUT2− mothers had no cases of ARI with fever (p=0.006). In this ongoing study, we plan to continue our analysis to include the infant’s own FUT2 status and medically‐attended illness.

Support or Funding Information

CDC grant number U191P001059, IP‐16‐004
A science mapping and scientometric study on the application of botanical agents to the research on testosterone
Authors: Sohn Eunsoo , Sohn Eun-Hwa
Published: April 2020

Many physiological and pharmacological studies on testosterone(TS), the major male sex hormone, have been carried out over the past decades. This study aimed to quantify the research trend based on TS scientific publications in a 23‐yr period, and provide the quantitative estimates of the TS research associated with botanical agents(BA) using scientometric methods. Total 2,279 bibliographic records retrieved from the SCOPUS database from 1996 to 2018 were analyzed with Knowledge Matrix Plus, VOS viewer and Gephi software. We obtained science map derived from co‐word network analysis and identified TS research area related to BA. The results revealed that TS studies related to BA have concentrated on the modulating effect of BA on TS and TS metabolite levels in environmental, medical, and toxicological research field. The findings report that TS is highly related to risk assessment factors such as endocrine disruption exposed to pollution and wastewater in industrial, agricultural and livestock environments. In medical studies, spermatogenesis, testis, erectile dysfunction, and fertility studies have been observed in TS research involving BA. Research on the development of 5‐alpha‐reductase inhibitor for alopecia, and the regulation of bcl‐2 for carcinogenesis were also revealed. In toxicological studies, side effects of TS therapy for prostate cancer, hyperplasia, hepatotoxicity (cytochrome p450), and lipid peroxidation were investigated in a relatively higher frequency.

Support or Funding Information

This research was supported by Korea Institute of Science and Technology Information (KISTI) under the research program “Development of Future Technology Analysis System Based on Open Data”.
Urine metabolomics and proteomics for bladder cancer prediction by LC/MS based strategy
Authors: Tang Xiaoyue , Guo Zhengguang , Sun Haidan , Liu Xiaoyan , Liu Xiang , Li Jing , Lian Xiaoxiao , Zheng Shuxin
Published: April 2020

Early diagnosis and life‐long surveillance are clinically important to improve the long‐term survival of bladder cancer (BCa) patients. Cystoscopy is considered the gold standard for the clinical diagnosis of human BCa. However, it is expensive and invasive. The objective of this study was to discover novel biomarkers to distinguish BCa from healthy controls, urothelium carcinoma and benign bladder lesions by LC‐MS based metabolomics and proteomics. We also tried to differentiate high‐grade and low‐grade BCa. A total of 234 subjects were enrolled, including 80 healthy adults, 22 benign patients, 26 urothelium carcinoma patients and 106 BCa patients (63 high‐grade BCa, 43 low‐grade BCa). The results showed that both urine metabolites and urine proteins could distinguish the BCa group from the healthy group, urothelium carcinoma group and benign bladder lesions group. And the combination of metabolites and proteins could achieve higher accuracy. Similarly, BCa clinical stages (high‐ vs. low‐grade BCa) could be better separated with a panel consisting of metabolites and proteins than either the metabolite panel or the protein panel alone. In conclusion, this study showed that urine proteomic and metabolic profiling were able to assist in the detection of BCa and grading distinguish.

Support or Funding Information

National Basic Research Program of China (No. 2013CB530805, 2014CBA02005)
An Initial Preparation for Cadaveric Dissection Ameliorates the Associated Mental distress of Human Cadaveric Dissection
Authors: Shaik Mustafa vali
Published: April 2020

Introduction

It is universally recognized that cadaveric dissection is an essential part of training in anatomy. However, it has also been reported to lead to mental distress in some students and can even impair their intrinsic motivation to study. One of the postulated reasons for this behavior is the lack of adequate information and preparation for cadaveric dissection. Hence it is hypothesized that providing relevant information prior to cadaveric dissection will ameliorate the mental distress of students, and improve in their academic performance.

Methods

A cohort of Masters of Occupational Therapy students enrolled in an Anatomy course were provided with a curated list of journal articles and videos related to cadaveric dissection, prior to the start of the course. All students were also required to attend an oral presentation related to cadaveric dissection, immediately before commencing of dissection. The control group consisted of students who had not been provided any information or prepared for cadaveric dissection.

Results

When compared to the control group, students who had been prepared for cadaveric dissection reported a lesser degree of mental distress. In addition, they also demonstrated greater intrinsic motivation, and better quality of cadaveric dissection. Moreover, students reported the oral presentation to be most relevant and journal articles to be least useful in this regard.

Conclusion

Hence preparation for cadaveric dissection is a valuable approach in the amelioration of mental distress and improvement of academic performance in anatomy students. Therefore, it is suggested that this approach could be a requisite in human anatomy.

Support or Funding Information

Joydeep D Chaudhuri Joydeep D Chaudhuri, MD Associate Professor, School of Occupational Therapy 314 Commons Husson University, 1 College Circle Bangor, ME 04401 Tel: 207 8528747 Email:
chaudhurij@husson.edu
Improved Sonic Hedgehog Protein Autoprocessing Assay in Cells Using Luciferase Reporter System
Authors: Ciulla Daniel , Mancusi Rebecca , Psaras Alexandra , Ghotra Sahib , Callahan Brian
Published: April 2020

Dysregulated cell signaling by Sonic hedgehog (Hh) proteins is implicated in developmental malformations and sporadic tumor growth. Biosynthesis of Sonic Hh requires cholesterolysis, a specialized autoprocessing event involving peptide‐bond cleavage and subsequent cholesterol modification. Cholesterolysis provides a potential site for targeted therapy. We are pursuing an in‐cell assay to monitor cholesterolysis activation and inhibition in multi‐well plates to improve on conventional techniques such as the Western Blot. Our chimeric reporter construct that fuses luciferase to human Sonic hedgehog cholesterolysis‐active domain (nLuc‐SHhC) appears to provide sensitive and quantitative measurement of cholesterolysis in HEK293 cells. Construct function was initially assessed by transient transfection. We are now preparing stable cell lines using CRISPR‐Cas9 system as a means for targeted gene insertion. Transient transfections demonstrate 5–10 fold difference in luciferase luminescence between catalytically active Sonic Hh and cholesterolysis‐defective mutant constructs. Wildtype construct yields a stronger luminescence in the supernatant, while the mutant Hh constructs show signal predominately in cell lysate. Additionally, stable cell lines show an even greater fold difference based on preliminary results. We intend to apply the reporter system introduced here for structure/function studies of cholesterolysis and for assessment of potential inhibitors targeting cholesterolysis.

Support or Funding Information

Funding for this research has been graciously provided by National Cancer Institute (Grant R01 CA206592) and Department of Defense (Grant W81XWH‐14‐1‐0155).
High‐Throughput Matrix Synergy Screening Identifies Novel Therapeutic Combinations for Merkel Cell Carcinoma
Authors: Garman Khalid Ammar , Gelb Tara , Urban Daniel , Hall Matthew David , Brownell Isaac Franklin
Published: April 2020

Merkel cell carcinoma (MCC) is a rare and aggressive neuroendocrine skin cancer. The majority of MCC tumors are virus‐positive (VP‐MCC) with Merkel cell polyomavirus integrated into the host genome, whereas virus‐negative (VN‐MCC) tumors are associated with mutations induced by ultraviolet light exposure. Primary MCC is treated by surgical resection and radiotherapy. For metastatic MCC, conventional chemotherapy is ineffective, and although immune checkpoint inhibitors (ICI) can produce durable responses in some cases, many MCC patients cannot receive ICI and others have disease progression despite immunotherapy. To identify novel treatments for MCC, we performed high‐throughput small molecule screening of ~4,000 FDA‐approved and experimental drugs for their ability to reduce MCC viability in VP‐MCC and VN‐MCC cell lines relative to immortalized control cell lines. Drugs were grouped by target and ranked according to area under dose‐response curve (AUC). We identified navitoclax as selectively efficacious against VP‐MCC cell lines compared to VN‐MCC and controls. However, navitoclax use in the clinic is limited by a dose‐dependent thrombocytopenia. To identify drug combinations that will lower the effective dose of navitoclax and thereby decrease the risk of thrombocytopenia, we performed a synergy screen combining navitoclax with each of the 1912 clinically relevant compounds in the NCATS Mechanism Interrogation PlatE (MIPE) library. This combination study revealed synergistic effects between navitoclax and multiple drug categories. We are now testing high priority drug combinations for toxicity and efficacy in pre‐clinical xenograft models of MCC. Collectively, our study has identified novel therapeutic drug combinations for MCC with the potential for enhanced efficacy and lower side effect profiles. By focusing on repurposing approved drugs, these combinations will be immediately available to patients who fail ICI therapy.

Support or Funding Information

NIH Intramural Research Program
Understanding the Interactions Between γ‐B Crystallins
Authors: Melake Abigail , Fraser Olivia , Fadden Aaron , Faraone Julia , Williams Zach , Mills Jeffrey , Thurston George , Michel Lea Vacca
Published: April 2020

The eye lens functions to focus light on the retina and is mostly composed of α, β, and γ‐crystallins, which are water‐soluble proteins. Aggregation of these proteins due to insolubility causes light to scatter, which can result in a cataract, leading to visual impairment or blindness. Aggregation of the crystallins can be due to unfavorable protein‐protein interactions. Here, we describe our analysis of some of those protein‐protein interactions using nuclear magnetic resonance (NMR) spectroscopy. Preliminary data suggest that NMR is a useful tool in exploring the detailed protein‐protein interactions between crystallins. Specifically, we used T1/T2 and DOSY NMR experiments to show that rotational and translational diffusion of bovine γ‐B crystallin proteins (homologue to human γ‐D crystallins) are affected by temperature and protein concentration and possibly changes in protein aggregation.
Interactions of Estrogen and TRPM5 Channels in Fat Taste and Metabolism
Authors: Dahir Naima , Calder Ashley , Gilbertson Timothy A.
Published: April 2020

Transient Receptor Potential channel type Melastatin 5 (TRPM5) is a nonselective cation channel that is activated by an increase in cytosolic calcium. TRPM5 channels are highly expressed in Type II taste receptor cells that transduce bitter, sweet, umami, and fatty acid compounds. While the function of TRPM5 channels is well‐defined in these cells, the effects of hormones such as estrogen on TRPM5‐mediated signaling that impact the taste system are not well known. We investigated TRPM5 channel activation in fatty acid‐sensing cells and the physiological impact in long‐term fat intake in males and females. This study followed from our earlier work where we showed a sex difference in TRPM5‐deficient (TRPM5−/−) animals and their wild type (WT) counterparts on a 60% high‐fat diet. We showed TRPM5−/− male mice were obese‐resistant compared to their WT counterparts. In comparison, TRPM5−/− females showed no drastic change in caloric intake but gained less body weight and body fat compared to the WT females. We found that removing the secretion of estrogen via ovariectomy in the TRPM5−/− females changed their weight status; they gained the same amount of weight and body fat as the WT females while not altering their caloric intake. At the taste cell level, we found that estrogen increased fatty acid‐induced Ca2+ responses in both TRPM5−/− and their WT counterparts. Interestingly, this difference was papillae‐specific in the TRPM5−/− females; circumvallate taste cells showed no increase in fatty acid responses when estrogen was present. Fungiform taste cells displayed a striking increase in cytosolic calcium in TRPM5−/− but not in WT females. Together, these results demonstrate a link between estrogen and TRPM5‐mediated signaling in feeding behavior and taste cells that is both sex and papillae‐specific. We are conducting additional experiments to better understand the mechanistic interactions between estrogen receptor activation and taste signaling via TRPM5 channels.

Support or Funding Information

Supported by NIH DC013194 and DC013318 (tag)
ASIC1a plays a key role in evoking the metabolic component of the exercise pressor reflex in rats
Authors: Ducrocq Guillaume P. , Kim Joyce S. , Estrada Juan A. , Kaufman Marc P.
Published: April 2020

The role of the acid‐sensing ion channel 1a (ASIC1a) in evoking the exercise pressor reflex is unknown despite the fact that ASIC1a is opened by decreases in pH in the physiological range. This fact prompted us to test the hypothesis that ASIC1a plays an important role in evoking the exercise pressor reflex in decerebrated rats with freely perfused hindlimb muscles. To test this hypothesis, we measured the effect of injecting two ASIC1a blockers into the arterial supply of the triceps surae muscles on the reflex pressor responses to four maneuvers, namely (1) static contraction of the triceps surae muscles (i.e., the exercise pressor reflex), (2) calcaneal tendon stretch, and (3) intra‐arterial injection of lactic acid and (4) diprotonated phosphate. We found that the two ASIC1a blockers, psalmotoxin‐1 (200 ng kg−1) and mambalgin‐1 (6.5 μg −1), decreased the pressor responses to static contraction as well as the peak pressor responses to injection of lactic acid and diprotonated phosphate. In contrast, neither ASIC1a blocker had any effect on the pressor responses to tendon stretch. Importantly, we found that ASIC1a blockade significantly decreased the pressor response to static contraction after a latency of at least 8 s. Our results support the hypothesis that ASIC1a plays a key role in evoking the metabolic component of the exercise pressor reflex

Support or Funding Information Funding for this study was provided by the National Institute of Arthritis and Muskuloskeletal and Skin Diseases (Grant R01‐AR‐059397) and National Heart, Lung, and Blood Institute (Grant P01‐HL‐134609).
Rab GTPases Accelerates GLUT4 Translocation in Colorectal Cancer Progression by Insulin/IGF System
Authors: Chen Chi-Long , Chang Yu-Chan , Hsiao Michael
Published: April 2020

Colorectal cancer is one of the high risks of clinical patients, especially its strong metastatic ability and drug resistance. The components of metabolites or metabolism‐related proteins are individually linked with the prognosis and survival rate of colorectal cancer, respectively. High expression of Insulin‐like growth factor‐1 (IGF‐1) and its receptor (IGF‐1R) in colorectal tumor tissues was studied. Meanwhile, glucose transporter 4 (GLUT4) is an insulin‐response transporter of glucose homeostasis in humans. Several studies reported that GLUT4 translocated from cytoplasm to cell membrane for further serial glycolysis process. Through colorectal tumor tissue array screening, we observed that all class I glucose transporters (GLUT1‐4) were overexpressed in tumors compared to adjacent normal tissues. Moreover, we confirmed that GLUT4 expression levels were highly correlated with IGF1/IGF‐1R by immunohistochemistry and were further associated with survival curve and other clinical parameters of colorectal cancer. We further determined the dynamic switching of GLUT4 is between GLUT4‐storage vesicles (GSVs) in cytoplasm with cell membranes through immunofluorescence assay. Our results indicated that several RABs (RAB8/10/14) bound to GSVs and accelerated GLUT4 translocation, so GLUT4 facilitated the efficiency of glucose transporters in colon cancer cell lines. Furthermore, hyperglycemic conditions prevented cell apoptosis of chemotherapy drug (5‐FU, oxaliplatin and irinotecan) resistance. Therefore, we utilized glucose transporter inhibitors as antagonists in combination with chemotherapy drugs. These results may have clinical implications for chemotherapy strategies to improve colorectal cancer.
HYPOXEMIA PREVENTS RECOVERY FOLLOWING FENTANYL OVERDOSE IN AWAKE AND SEDATED RATS
Authors: Guck Daniel , McCann Marissa , Tubbs Nicole , Haouzi Philippe
Published: April 2020

Background

As severe acute hypoxemia produces a rapid inhibition of the respiratory neuronal activity through a non‐opioid mechanism, we have investigated in adult rats the effects of hypoxemia following fentanyl overdose‐induced apnea on 1‐ auto‐resuscitation and 2‐ the antidotal effects of naloxone.

Methods

In unsedated rats, the breath‐by‐breath ventilatory and pulmonary gas exchange response to fentanyl overdose (300 mg.kg−1.min−1 iv in one minute) was determined in an open flow plethysmograph. The effects of inhaling air (9 rats) or a hypoxic mixture (FIO2 between 7.3 amd 11.3%, 8 rats) on the ability to recover a spontaneous breathing rhythm and on the effects of naloxone (2 mg/kg) were investigated. In addition, arterial blood gases, arterial blood pressure, ventilation and pulmonary gas exchange were determined in spontaneously breathing tracheostomized urethane‐anesthetized rats in response to 1‐ fentanyl induced hypoventilation (7 rats), 2‐ fentanyl induced apnea (10 rats) in air and hyperoxia, and 3‐ isolated anoxic exposure (6 rats).

Results

Data are expressed as median and range. In the unsedated rat model, bolus fentanyl injection in air breathing produced an apnea within 14 seconds (12 – 29 seconds). A spontaneous rhythmic activity resumed after 85.4 s (33–141 seconds) consisting in a persistent low tidal volume and slow frequency rhythmic activity that rescued all animals. Naloxone at 10 minutes immediately restored the baseline level of ventilation. At FIO2 below 10%, fentanyl induced apnea was irreversible despite a transient gasping pattern, the administration of naloxone had no effects. In sedated rats, no spontaneous recovery of breathing occurred following fentanyl‐induced apnea, when PaO2 was lower than 16 mmHg, akin to the effects of hypoxemia alone. Naloxone had no rescuing effect on fentanyl‐induced apnea despite circulation being maintained.

Conclusion

Hypoxia‐induced ventilatory depression during fentanyl induced apnea 1) opposes the spontaneous emergence of a respiratory rhythm, which would have rescued the animals otherwise, 2) prevents the effects of high dose naloxone.

Support or Funding Information

None
Interferon‐γ Induces PD‐L1 Expression in Ovarian Cancer Cells by JAK/STAT1 Signaling
Authors: Padmanabhan Sveta , Gaire Bijaya , de Leon Daniel , Vancura Ales , Vancurova Ivana
Published: April 2020

Interferon‐γ (IFNγ) is a pleiotropic cytokine critical for immune functions and tumor immunity. Although IFNγ has been used in the treatment of different types of cancer, including ovarian cancer (OC), most clinical trials using IFNγ to treat solid tumors have produced mixed or disappointing results. Recent studies have shown that IFNγ induces expression of the immune checkpoint PD‐L1 in OC cells, resulting in their increased proliferation and tumor growth. However, the mechanisms that regulate the PD‐L1 expression in OC remain poorly understood. Since one of the main pathways induced by IFNγ is the JAK/STAT pathway, we investigated whether the IFNγ‐induced PD‐L1 expression in OC cells is regulated by JAK/STAT signaling. Here, we show that IFNγ induces STAT1 expression and STAT1 phosphorylation on Tyr‐701 and Ser‐727 in OC cells. In addition, JAK1 suppression significantly decreases the IFNγ‐induced PD‐L1 mRNA and protein levels in OC cells, demonstrating that the IFNγ‐induced PD‐L1 expression in OC cells is dependent on JAK1 activity. Together, our results indicate that the IFNγ‐induced PD‐L1 expression in OC cells is regulated by JAK/STAT1 signaling. Understanding the mechanisms of how IFNγ induces the PD‐L1 expression in OC cells will contribute to the development of therapeutic strategies aimed to increase the anti‐tumor effects of IFNγ, and modulate the levels of PD‐L1 in ovarian cancer.

Support or Funding Information

NIH CA202775 grant
Choice of Hunting Method Influenced Human‐Carnivore Competition in Late Pleistocene Southeast Asia
Authors: Zachwieja Alexandra J , Bacon Anne-Marie , Nguyen Thi Mai Huong , Nguyen Anh Tuan , Sichanthongtip Phonephanh , Sayavongkhamdy Thongsa , Demeter Fabrice , Shackelford Laura L
Published: April 2020

Competition has been investigated as a source of dispersal pressure in animal species both past and present. Particularly in Southeast Asia, where the Pleistocene carnivore guild remains largely intact to this day, there is a unique opportunity to assess the effects of competition on both dietary and spatial (dispersal) niche partitioning behavior in humans. Here, we investigate the relationship of early modern humans to local carnivores during their likely period of dispersal through Laos and Vietnam (~80‐50ka) to – for the first time – assess humans as a member of a local carnivore guild. By using the Competition Index (CI), a method which has shown success in predicting competitive pressure in island Southeast Asian paleontological communities on a scale of 0 (no competition) to 1 (full prey overlap, maximum competition). We approach this research with a purposeful desire to remove the “bias of human exceptionalism” from our analysis, treating humans as another medium size animal. We find that humans definitively were subject to competitive pressure from concurrent carnivores, particularly medium‐size prey specialists like hyenas, dholes, and leopards (Max CI: 0.6, 0.5, 0.66 respectively). Additionally, we find that the estimated competitive pressure on humans changes accounting for hunting method (solo ambush vs. group hunting), with group hunting practices reducing competitive pressure for medium‐size prey, particularly from leopards. We suggest that hunting behavior may have been used along with dietary and spatial partitioning as a strategy to reduce competitive pressure during initial human dispersal in Southeast Asia and that biotic interactions should not be overlooked in studies of human dispersal in this region.

Support or Funding Information

Funding for this work was provided by: National Geographic Early Career Grant #EC‐378R‐18, Wenner‐Gren Foundation Dissertation Fieldwork Grant, UIUC Anthropology Department, UIUC Graduate College, The Explorer’s Club Exploration Fund, and the Beckman Institute CS/AI Award.
Knockdown and Inhibition of SYK Tyrosine Kinase Decreases mTORC1 Activity and Enhances Lysosomal Biogenesis and Autolysosomal Maturation in Pancreatic Cancer Cell Lines
Authors: Pan Michelle , Hua Kevin , Villait Akash , Zuger Angelina , Rafati Minoo , Chen Ryan , Liu Sherry , Singh Anurag
Published: April 2020

In cancer, oncogene dependency is a phenomenon where a dominant driver oncogene promotes tumor cell proliferation and survival, and loss of this oncogene results in tumor cell death and, eventually, tumor regression. KRAS, a GTPase that regulates cell growth and proliferation, is an oncogene constitutively activated in over 90% of pancreatic cancer. However, clinically effective inhibitors of KRAS have been unsuccessful so efforts have been focused on identifying other potential targets associated with the KRAS signaling network. Spleen tyrosine kinase (SYK), expressed at high levels in KRAS‐dependent pancreatic cancer cell lines, may be one of these targets. Our data indicate that SYK activates the mechanistic target of rapamycin kinase complex 1 (mTORC1), which promotes protein translation and cell growth. SYK activation also leads to decreased autolysosome count. In connecting SYK activation with increase mTORC1 activity and decrease autolysosome count, we hypothesis that the MiT/TFE transcription factors are involved. We propose that SYK inhibition in pancreatic cancer cells leads to reduced mTORC1 activity, which reduces phosphorylation of MiT/TFE transcription factors. The unphosphorylated MiT/TFE transcription factors are then allowed into the nucleus to activate genes for lysosomal biogenesis and autophagy. Autophagy is a process that recycles cellular macromolecules during nutrient deprivation by fusing autophagosomes and lysosomes, produced from lysosomal biogenesis, to generate autolysosomes. From our experiments, we were able to show that SYK inhibition blocks mTORC1‐dependent phosphorylation of MITF and TFEB transcription factors, members of the MiT/TFE family. MITF and TFEB activation leads to increased autophagy due to autolysosomal biogenesis and accumulation. In summary, our studies of the SYK‐mTORC1‐autophagy pathway provide support to investigate SYK as a candidate, therapeutic target for pancreatic cancer treatment.

Support or Funding Information

NIH/NCI K99/R00 ‐ Anurag Singh (PI); Boston University Dahod Breast Cancer Pilot Award ‐ Anurag Singh (PI); Boston University's Undergraduate Research Opportunities Program ‐ Michelle Pan
NLRP3 inflammasome activation in dopamine neurons contributes to neurodegeneration in Parkinson’s Disease
Authors: Panicker Nikhil , Kam Tae-In , Neifert Stewart , Hinkle Jared , Mao Xiaobo , Karuppagounder Senthilkumar , Wang Hu , Kumar Manoj , Pirooznia Sheila , Pletnikova Olga , Troncoso Juan , Dawson Valina L. , Dawson Ted M.
Published: April 2020

Parkinson’s disease (PD) is a progressive neurodegenerative disorder that affects over a million people within the U.S. Underlying the clinical symptoms of PD is the degeneration of dopamine (DA) neurons located in the pars compacta of the substantia nigra (SNpc). Microglial activation of the NLRP3 inflammasome has been well‐documented in various neurodegenerative diseases, including PD. We report herein that adult onset conditional knockdown of Parkin in the mouse SNpc results in priming and activation of the NLRP3 inflammasome within dopamine neurons. These results were replicated in Parkin knockout generated human embryonic stem cell (hES) differentiated DA neurons (hDA neurons). Next, we utilized Nlrp3A350VneoR mice to demonstrate that dopamine neuron specific activation of the NLRP3 inflammasome is sufficient to elicit DA neuron death. Double transgenic Parkinflx/flx/Casp1KO mice were protected from adult onset Parkin deletion‐mediated DA neuron death. Finally, we demonstrate that Parkin is inactivated in alpha‐synuclein pre‐formed fibril (PFF)‐treated cortical neurons and in‐vivo, licensing neuronal NLRP3 inflammasome activation in sporadic PD mouse models as well. Taken together, our results indicate that atypical NLRP3 inflammasome activation within DA neurons may constitute a novel cell‐death mechanism that may contribute to neurodegeneration in PD.

Support or Funding Information

The first (and presenting) author is supported by a post‐doctoral fellowship from the Maryland Stem Cell Research Fund (MSCRF) Grant ID: 2017‐MSCRF‐3838
PAPSS2 ablation prevents acetaminophen‐induced hepatotoxicity in mice
Authors: Xu Pengfei , Wang Pengcheng , Xi Yue , An Yunqi , Xu Meishu , Ren Songrong , Ma Xiaochao , Xie Wen
Published: April 2020

Sulfate plays an essential role in numerous biochemical and cellular processes in mammalian physiology. The PAPS synthase 2 (PAPSS2) is the primary enzyme to synthesize 3'‐phosphoadenosine 5'‐phosphosulfate (PAPS), which is the universal sulfonate donor for all sulfonation reactions in mammals. Acetaminophen (APAP) overdose is the leading cause of acute liver failure in the United States, and sulfation is widely believed to contribute to the detoxification of many drugs including APAP. The goal of this study is to determine whether PAPSS2 plays a role in APAP‐induced hepatotoxicity. Surprisingly, we found the PAPSS2 knockout mice were protected from APAP‐induced hepatotoxicity, and this protection was accompanied by increased antioxidant ability through the activation Nrf2‐Keap1 and p53 pathways. The hepatoprotective effect of PAPSS2 ablation was abolished when p53 was knocked down. In summary, we have uncovered a unique role of PAPSS2 in APAP‐induced hepatotoxicity.

Support or Funding Information

This work was supported in part by National Institutes of Health Grants DK083952, ES023438
Characterizing of the Effects of Pal‐Peptidoglycan Interaction
Authors: Liu Xinbei , Lewis Sean , Ward Zack , Michel Lea Vacca
Published: April 2020

Peptidoglycan‐associated lipoprotein, Pal, is found in the outer membrane of Escherichia coli (E. coli) and other Gram‐negative bacteria. Previous research has shown that under certain conditions, including several animal models of sepsis, Pal is released from E. coli and is thought to be implicated in the clinical condition of sepsis. In Gram‐negative bacteria, Pal is non‐covalently, but tightly bound to peptidoglycan. We hypothesize that the strong Pal‐peptidoglycan interactions influence Pal’s ability to be released from E. coli. Site‐directed mutants of E. coli Pal have been prepared and studied to determine how Pal’s altered binding affinities to peptidoglycan affect Pal’s release from E. coli. Preliminary results suggest that other factors, in addition to the Pal‐peptidoglycan interaction, are responsible for Pal’s release from E. coli.
LPAR3 Is Associated with Migration of NIH 3T3 Cells and Progression to Malignant Cell Transformation during Carcinogenesis
Authors: Lee Michael , Hwang Sung-Hee , Yeom Hojin
Published: April 2020

We have previously identified that the LPAR3 gene is commonly hypermethylated in foci transformed by two non‐genotoxic carcinogens (NGTxC). In this study, we validated the down‐regulation of LPAR3 genes using qPCR analysis in both NGTxC‐induced transformed foci and another v‐Ha‐ras‐transformed NIH 3T3 cells (Ras‐NIH 3T3). We revealed that LPAR3 overexpression significantly increased migration of Ras‐NIH 3T3 whose migration capacity was much lower than that of their parental cells. However, LPAR3 overexpression in Ras‐NIH 3T3 cells resulted in the inhibition of cell survival concomitant with G2/M arrest. On the other hand, inhibition of LPAR3 signaling with Ki16425 had little effect on cell survival of Ras‐NIH 3T3 cells with low level of LPAR3 expression, whereas inhibiting cell survival of NIH 3T3 cells. These results imply that LPAR3 plays a positive role in proliferation of NIH 3T3 cells but not in that of its ras‐transformed cells. To further analyze the role of LPAR3 during carcinogenesis, LPAR3 knockout (KO) in NIH 3T3 cells was generated using the CRISPR/Cas9 genome editing. Four clones from nine initial clones were confirmed with sequencing for genomic editing at the target site. From this initial screen, one clone with biallelic homozygous fragments deletions was selected for further analysis. WST‐1 assay showed that LPAR3 KO greatly inhibited cell survival of NIH 3T3 cells. LPAR3 KO also reduced migration of NIH 3T3 cells. Taken together, our results indicate that LPAR3 might be functionally involved in malignant cell transformation. Further research will be required to understand the intrinsic relationships between cell survival and migration.

Support or Funding Information

This work was supported by the National Research Foundation of Korea (NRF) grant funded by the Ministry of Science and ICT (MIST) (NRF‐2019R1F1A1048733).
Variation in the origin of the first motor branch of the ulnar nerve to the flexor carpi ulnaris muscle
Authors: Dugan Sonja Hedblom , Lundgren Mary , Inouye Sandra , Molen Jonathan Vander , Freemyer-Brown Anna
Published: April 2020

Introduction/Purpose

Surgical decompression of the ulnar nerve in the treatment of cubital tunnel syndrome is performed in close proximity to the first motor branch of the ulnar nerve to flexor carpi ulnaris (FCU). Understanding the variability of this branch is important for improving surgical interventions. The purpose of this study was to determine the location and anatomical variation of the first motor branch to flexor carpi ulnaris.

Methods

Embalmed cadavers were dissected to expose the ulnar nerve as it courses past the medial epicondyle of the humerus. Digital calipers were used to measure the distance between the point of origin of the first motor branch of the ulnar nerve to FCU and the most prominent aspect of the medial epicondyle. The length of the ulna was also measured and correlated with data from ulnar nerve measurements. Statistical significance was defined as a p‐value < 0.05.

Outcomes

Data was obtained from 113 cadaveric elbows. The location of the branch point was most consistently found (88.5% of specimens) distal to the medial epicondyle (average 19.42 mm, standard deviation 4.39, 95% confidence interval 18.54 – 20.29). In 11.5% of elbows, however, the branch point occurred proximal to the medial epicondyle (average 16.02 mm, standard deviation 4.53, 95% confidence interval 13.28 – 18.76 mm). The average distance was 18.42 mm distal to the medial epicondyle in females and 20.45 mm in males, with a statistically significant difference between genders (p‐value = 0.02). The correlation between the branch point of the first motor branch of the ulnar nerve to FCU and ulna body size was also explored.

Conclusion

This study has demonstrated that the first motor branch of the ulnar nerve is most consistently located 18.54 to 20.29 mm distal to the medial epicondyle. Males had a significantly greater distance from the origin of the first motor branch of the ulnar nerve to FCU from the medial epicondyle compared to females. No statistically significant correlation was found related to laterality. Additionally, the prevalence of the anatomical variant in which branching of the first motor nerve from the ulnar nerve occurs proximal to the medial epicondyle was estimated. Prior to this study, the prevalence of this variant had not been reported.

Support or Funding Information

The authors would like to thank Midwestern University for the use of the specimens in this study.