bluuebunny/crossref_metadata_2025 huggingface dataset:
Zinc Deficiency‐Induced Renal NFkB Activation drives Sodium Chloride Co‐transporter Upregulation.
Authors: Wenegieme Tara-Yesomi , Waite Aston , Naraine Meagan , Murta Cindellynn , Schindele Dylan , Sonner Martha , Williams Clintoria
Published: April 2020

Background

Zn2+ deficiency (ZnD) is a worldwide problem. In the United States, 14% of Americans are Zn2+ deficient, which represents 1 out of 7 people. In ZnD populations, the prevalence of hypertension is higher. In our recent studies, we demonstrated that ZnD induces hypertension by promoting renal Na+ reabsorption by the sodium chloride co‐transporter (NCC). However, the exact molecular mechanisms involved in NCC upregulation were undefined. Nuclear factor‐κB (NFκB) is a transcription factor found to play a role in ZnD‐mediated detrimental effects throughout the body.

Hypothesis

As such, we hypothesized that ZnD drives renal NFκB activation.

Experimental Design

To examine the effects of ZnD on renal NFκB activation, adult, male C57Bl/6 mice were fed a ZnA‐ or ZnD‐diet for 6 weeks. NFκB expression and nuclear translocation were examined by immunohistochemistry. To confirm the role of Zn2+ in NFκB regulation, mouse distal convoluted tubular (mDCT) cells were treated with the Zn2+ chelator ‐ N,N,N′,N′‐tetrakis(2‐pyridylmethyl)ethane‐1,2‐diamine (TPEN) or vehicle ± Zn2+ supplementation.

Results

In ZnD mice, NFκB protein expression and nuclear localization were increased compared to ZnA mice. Consistently, in mDCT cells, TPEN‐induced ZnD stimulated NFκB expression and nuclear translocation. However, Zn2+ supplementation reversed TPEN‐induced NFκB upregulation.

Conclusion

These results indicate that 1) NFκB is a Zn2+‐regulated nuclear transcription factor, and 2) ZnD drives renal NFkB activation.

Significance

NFκB represents a potential mediator that drives ZnD‐induced NCC upregulation and consequently renal Na+ reabsorption and hypertension.

Support or Funding Information

American Heart Association ‐ Scientist Development Grant #16SDG27080009 National Institutes of Health ‐ R21 Exploratory/Development Grant #R21DK119879
Effect of Vancomycin on Cytoplasmic Peptidoglycan Intermediate Levels in Vancomycin Resistant Enterococci.
Authors: Gargvanshi Shivani , Gutheil William
Published: April 2020

Vancomycin is a natural glycopeptide antibiotic and is used for the treatment of multi‐drug resistant Gram positive bacteria. However, resistance to vancomycin has emerged. Vancomycin resistance in VanA‐type Enterococcus faecium is due to a vancomycin resistance cassette comprised of vanH, vanX, vanY, vanZ, vanR and vanS genes. These genes encode for an alternative peptidoglycan biosynthesis pathway in which D‐Ala‐D‐Ala is replaced with D‐Ala‐D‐Lac. Vancomycin binds with a lower affinity to D‐Ala‐D‐Lac as it does to the nomal dipeptide. In this study, several of the unique UDP‐linked intermediates from VanA‐type vancomycin resistant Enterococcus faecium (VRE) were preparatively purified from vancomycin treated cultures and used to optimize their LC‐MS/MS quantification. Cytoplasmic pool levels were then measured in VRE before and after vancomycin exposure. VRE in the absence of vancomycin had low but readily detectible basal levels of VanA pathway intermediates. After a 15 min vancomycin exposure, normal pathway intermediates were only modestly affected, whereas alternative pathway intermediates increased substantially. Alternative pathway intermediates then decreased back toward unexposed levels after longer vancomycin exposure. A vancomycin concentration profile demonstrated a midpoint of vancomycin effect between 1 and 4 μg/mL. Time course experiments revealed a complex response to vancomycin exposure, with most normal pathway intermediates responding rapidly (minutes) to vancomycin exposure, and alternative pathway intermediates increasing for 15–20 min and then decreasing partially back toward normal levels. qRT‐PCR study also demonstrated that mRNA levels of VanA pathway genes also took about 15 min to reach their maximal level of induction.

Support or Funding Information

NIH R15
Radioresistant Deinococcus Actinosclerus BM2T Suppresses Lipopolysaccharide‐mediated Inflammation in the RAW264.7 Cells
Authors: Park Yuna , Jang Seon-A , Lee Jin Woo , Kim Sung Hyeok , Ha Chang Woo , Lee Sung Ryul , Kim Myung Kyum , Sohn Eun-Hwa
Published: April 2020

Deinococcus actinosclerus BM2T (designated to KT448814) is newly isolated radioresistant bacterium from soil of a rocky hillside. As an extremophile, D. actinosclerus BM2T may express health‐beneficial compounds for combating inflammation. In this study, we evaluated anti‐inflammatory potentials of BM2W, a water extract of D. actinosclerus BM2T, on lipopolysaccharide (LPS)‐mediated inflammatory responses in the RAW264.7 macrophages. LPS‐induced mRNA expression levels of proinflammatory cytokines, such as interleukin (IL)‐1β, IL‐6 and tumor necrosis factor‐α, were significantly inhibited in the presence of BM2W. Inductions of inflammatory mediator producing inducible nitric oxide synthase and cyclooxygenase‐2 proteins were also suppressed by BM2W treatment. BM2W led inhibition of activation of nuclear factor‐κB and mitogen‐activated protein kinases (JNK, ERK and p38). Collectively, BM2W seems to be potent in attenuating inflammatory responses via MAPKs/NF‐κB pathway.
Filament Formation Enables Highly Cooperative Regulation of Human CTP Synthase 2
Authors: Lynch Eric , Kollman Justin
Published: April 2020

CTP synthase is one of a variety of metabolic enzymes recently found to polymerize into large‐scale filaments. CTP synthase catalyzes the ATP‐dependent conversion of UTP to CTP, and is also allosterically regulated by GTP, making it sensitive to levels of all four major ribonucleotides, and therefore an important regulatory node in nucleotide metabolism. For many filament‐forming enzymes, polymerization is driven by binding to substrates, products, or allosteric regulators, and modulates activity by trapping the enzyme in either a high or low activity conformation. Here, we describe a unique, ultrasensitive mechanism of regulation in human CTP synthase 2 (CTPS2) filaments. High‐resolution cryo‐EM structures reveal that rather than selectively stabilizing a single conformation, CTPS2 filaments switch between active and inhibited conformations in response to changes in substrate and product levels. By coupling the conformational state of many CTPS2 subunits within a filament, polymerization greatly enhances the cooperativity of CTPS2 regulation, allowing for more sensitive regulation compared with the CTPS2 tetramer alone. Our cryo‐EM structures of active and inhibited CTPS2 filaments also provide new insight into the mechanism controlling substrate transfer between the enzyme’s two active sites, and further reveal a novel mode of product binding. The ultrasensitive mode of regulation observed in CTPS2 filaments may aid in understanding the function of other enzyme filaments; the coordinated propagation of conformational changes along filaments could provide a mechanism for fine‐tuning the regulation of other polymerizing enzymes.
Western diet promotes renal injury, inflammation, and fibrosis in a murine model of Alström syndrome
Authors: Kim Young Chul , Ganguly Souradipta , Nespoux Josselin , Freeman Brent , Zhang Haiyan , Brenner David , Dhar Debanjan , Vallon Volker
Published: April 2020

Alström syndrome is a rare recessive genetic disease caused by mutations in ALMS1, which encodes a protein that is related to cilia function and intracellular endosome trafficking. The syndrome has been linked to impaired glucose metabolism and chronic kidney disease (CKD). Polymorphisms in ALMS1 have likewise been linked to CKD, but little is known about the modification of the phenotype by environmental factors. To gain further insights, the fat aussie (foz) mouse strain, a genetic murine model of Alström syndrome, was exposed to a normal chow (NC) or to a Western diet (WD, 20% fat, 34% sucrose by weight and 0.2% cholesterol). Body weight was 50% higher in foz vs WT mice on NC, and WD enhanced the difference to 64% (Pinteraction=0.0002, two‐way ANOVA). Kidney weight to body weight ratio was not different between groups. Plasma creatinine was lower in foz vs WT mice independent of diet, potentially indicating glomerular hyperfiltration in foz mice. Renal mRNA expression of injury marker Kim1 was not affected by WD in WT and was not different between foz and WT mice on NC (P=0.9995), but it was 26‐fold higher in foz vs WT mice on WD (P<0.0001). Likewise, other renal injury markers (Ngal mRNA; histology assessment) and renal markers of inflammation (Tnf, Il6, Il10 and Ccl2 mRNA; CD68‐positive cells) and fibrosis (Col1A1, Timp1 and Tgf‐beta mRNA; Masson’s trichrome staining) were similar in both genotypes on NC, but significantly greater in foz vs WT mice fed a WD. Albuminuria was higher in foz vs WT mice on both diets but WD significantly increased the difference (Pinteraction=0.0019, two‐way ANOVA). A glucose tolerance test performed in foz and WT mice on a WD revealed similar basal blood glucose levels and subsequent blood glucose profiles. These results indicate that a Western diet sensitizes a murine model of Alström syndrome to kidney injury, inflammation and fibrosis, an effect that may not be solely due to effects on glucose metabolism. Polymorphisms in ALMS1 may induce CKD in part by modulating the deleterious effects of high dietary fat and sucrose on the kidney.

Support or Funding Information

The authors were supported by NIH grants R01DK112042, R01DK106102 (VV), the UAB/UCSD O’Brien Center of Acute Kidney Injury NIH‐P30DK079337 (VV), the Department of Veterans Affairs (VV), and by an investigator‐initiated research project by Janssen (DB, VV). DD was supported by NIH/CTRI 1KL2TR001444 and the American Liver Foundation (ALF) “Liver Scholar” grants.
Lysine Acetylation in Granulosa Cells of PCOS Women Regulated by Carbohydrate Metabolism
Authors: Pei Wendi , Min Zheying , Yu Yang
Published: April 2020

Context

Lower‐quality embryos are often found in Polycystic Ovary Syndrome (PCOS) women during assisted reproductive technology treatment; however, there is still a lack of a reliable marker to predict the outcomes of fertilization and embryo development in the clinic.

Objective

The aims of this study were to comparatively profile protein lysine acetylation in ovarian granulosa cells, to understand its potential effect on ovarian function at the posttranslational modification level and to establish a reliable marker to predict embryo quality in women with PCOS.

Design

This study recruited 47 PCOS patients and 55 non‐PCOS controls. Mural granulosa cells were collected during oocyte retrieval surgery. A quantitative analysis of acetylated proteomics was carried out by mass spectrometry. The clinical presentations and outcomes of the patients with different acetylation levels in their granulosa cells were analyzed.

Results

In the granulosa cells, there was widespread lysine acetylation of proteins, of which 265 proteins had increased levels of acetylation and 68 proteins had decreased levels of acetylation in the PCOS group. Most notably, differentially acetylated proteins were significantly enriched in the metabolic pathways of glycolysis/gluconeogenesis, fatty acid degradation and amino acid biosynthesis. In addition, women with PCOS with enhanced acetylation in their granulosa cells had markedly reduced 2 pronuclear (2PN) rates, numbers of available embryos and rates of available embryos.

Conclusion

The dynamic levels of lysine acetylation in the granulosa cells of women with PCOS might alter ovarian metabolic homeostasis and can be a marker to reflect embryo quality in the clinic.

Support or Funding Information

This work was supported in part by the National Key R&D Program of China (2016YFC1000601) and the National Natural Science Funds (81571400, 81771580, and 81971381).
Directionally Opposite Effects Of Cyclosporine And Sirolimus On Endotoxic Nephrotoxicity In Rats
Authors: Elzokm Shrouk S. , Fouda Mohamed A. , Abdel Moneim Rahab A. , El-Mas Mahmoud M.
Published: April 2020

Immunosuppressant therapies have been shown to protect against inflammation, cardiovascular anomalies, and enhanced mortality during endotoxemia. The current study investigated whether end organ damage caused by endotoxemia is distinctly affected by calcineurin‐dependent (cyclosporine) and independent (sirolimus) immunosuppressants in rats. Biochemical, molecular, and histopathological studies were undertaken to determine the influence of simultaneous exposure to either immunosuppressant on renal and cardiac damage induced by endotoxemia. Serum biomarkers of kidney (urea, creatinine), but not heart (CPK), functions were significantly elevated in rats after 6 hr of treatment with lipopolysaccharide (LPS, 3 mg/kg i.p.). Moreover, LPS caused (i) vacuolation and degeneration of renal tubules, and (ii) significant increases in the width of proximal and distal tubules as appeared from morphometric analysis. Whereas these biochemical and histopathological influences of LPS were intensified upon concurrent exposure to cyclosporine (10 mg/kg i.p.), they were virtually eliminated after co‐treatment with sirolimus (1 mg/kg i.p.). Western expression studies demonstrated that increases in renal protein expressions of toll‐like receptor 4, monocyte chemoattractant protein‐1, and NADPH oxidase 2 were all potentiated and inhibited by cyclosporine and sirolimus, respectively. Unlike kidney data, the assessment of serum CPK or heart morphology revealed no evidence of cardiac abnormalities in rats treated with individual or combined LPS/immunosuppressant regimens. Together, the data underscore the capability of sirolimus, but not cyclosporine, to offset inflammatory and kidney damage in endotoxic states.
Role Of SDF:CXCR4 In Diet‐Induced Cardiac Injury
Authors: Dong Feng , Patel Karishma , Kiedrowski Matthew , Penn Marc
Published: April 2020

Introduction

Diabetic cardiomyopathy (DCM) is a major cardiovascular complication in patients with diabetes and is defined as ventricular dysfunction (in diabetes) independent of coronary artery disease. In this study, we define a novel role for the SDF‐1: CXCR4 axis in diabetes‐associated myocardial dysfunction.

Methods

Wild‐type mice were randomly assigned to a high fat high sugar diet (HFHS) or control diet (LF) for 14 months. Serial echocardiography was used to assess cardiac function. The hSDF‐1 plasmid was injected into the LV wall of HFHS mice 7 months and 14 months after HFHS diet. The mRNA levels in the hearts were quantified by qPCR.

Results

HFHS‐fed mice (vs. control diet) showed significantly increased deceleration time (diastolic dysfunction) 7 months after HFHS diet and decreased Ejection Fraction (EF) (systolic dysfunction) 14 months after the HFHS diet. We observed a significant increase in cardiac myocytes surface area and a decrease in vascular density in the hearts of HFHS mice. HFHS‐fed mice showed decreased P‐selectin, E‐selectin expression and increased CXCR4, Resistin, and DDR2 expression in the heart. There is no significant improvement in diastolic function after the SDF‐1 injection at 7 month. However, direct myocardial injection of hSDF‐1 plasmid led to a significant improvement in EF compared to the control group. To determine the role of CM CXCR4 in hyperglycemia associated cardiac injury, we quantified cardiac function and survival rate in HFHS‐fed CM‐CXCR4 null mice. We observed a significantly increased mortality rate before 14 months on HFHS fed CM‐CXCR4 null mice compare to HFHS‐fed control mice.

Conclusion

HFHS diet induces diastolic dysfunction in the short term then systolic dysfunction after a long term. SDF‐1 treatment ameliorates systolic dysfunction but not diastolic dysfunction. Our data suggest that cardiac myocyte expression of CXCR4 has an important role in diabetes‐associated cardiac injury.
Expression of Kv11.1 Channels in Lungs and Role in Pulmonary Hypertension
Authors: Shults Nataliia V. , Rybka Vladyslava , Suzuki Yuichiro , Brelidze Tinatin
Published: April 2020

Background

While Kv11.1 potassium selective channels are well known for their contribution to the heart repolarization little is known about their importance for the lung function. Here we show that Kv11.1 channels are abundantly expressed in the human and rat lung tissues.

Methods & Results

We found that, in the lungs of healthy humans and rats, Kv11.1 channel expression was limited to the large pulmonary arteries (PAs). In the lungs of human patients with chronic obstructive pulmonary disease (COPD) and of rats with pulmonary arterial hypertension (PAH), Kv11.1 channels were expressed not only in the large PAs, but also in the small PAs with the diameter <100 micrometers. The increase in the Kv11.1 channel expression followed the time course of the development of pulmonary vascular remodeling in PAH rats. Administration of dofetilide, a Kv11.1 blocker, to rats with PAH inhibited PAH‐associated pulmonary vascular remodeling.

Conclusions

Our study indicates that Kv11.11 channels are expressed in the lung and that these channels mediate the development of pulmonary vascular remodeling. Dofetilide is an FDA approved drug used for treatment of atrial fibrillation. Thus, our findings raise a possibility of repurposing this antiarrhythmic drug for the treatment of pulmonary hypertension.

Support or Funding Information

Supported by NIH
LC‐MS‐based urine metabolomics analyses for early diagnosis of medulloblastoma
Authors: Li Jing , Hao Xiaolei , Guo zhengguang , Sun Haidan , Liu Xiaoyan , Zhang Yang , Zhang liwei , Tian Yongji Yongji , Sun Wei
Published: April 2020

(MB) is the most common type of brain cancer in pediatric patients. In present study, we applied liquid chromatography ‐ mass spectrometry (LC‐MS) based metabolomics to analyze a cohort including 425 urine samples (118 healthy controls, 112 medulloblastoma patients, 33 patients with brain malignant cancers and 53 patients with brain benign diseases, 29 MB patients with surgery after a week and 80 MB patients with surgery after a month). Differential metabolites were analyzed to investigate if urine metabolites could differentiate MB from non‐MB. Apparent separation was observed between MB urine samples and healthy controls, brain benign diseases, other brain malignant tumor. The AUC in discovery group respectively were 0.94, 0.938, 0.808, and the AUC of external validation dataset were 0.913, 0.889, 0.822. According to the comparisons, 14 metabolites were found to be the specific metabolites of MB. Among 14 metabolites, two metabolites were validated in postoperative samples and with high AUC values (0.858 compare to non‐MB). Our data showed that urine metabolomics could be used for MB diagnosis and would provide candidates for the potential biomarkers of MB.

Support or Funding Information

National Basic Research Program of China (No. 2013CB530805, 2014CBA02005)
Protein‐Tyrosine Kinase, Syk Accelerates Phagosome Maturation by Depolymerizing F‐Actin in Complement‐Mediated Phagocytosis
Authors: Tabata Hiroyuki , Tohyama Yumi
Published: April 2020

Syk is a non‐receptor protein‐tyrosine kinase and primarily expressed in hematopoietic cells. We previously reported that Syk plays an essential role in complement‐mediated phagocytosis, especially in engulfment process. In the current study, we aimed at clarifying the role of Syk in phagosome maturation process in complement‐mediated phagocytosis. To reveal the role of Syk in the phagocytosis, we established various Syk‐knockout (Syk‐KO) cell lines by using CRISPR/Cas9 system in HL60 cells. Using these parental and mutant HL60 cells, we performed complement‐mediated phagocytosis assay after macrophage‐like differentiation. As a result, we found that Syk‐KO macrophages permitted escape of C. albicans from the phagosomes because of the insufficient phagosome acidification. Additionally, we found that fusion of phagosomes with lysosomes did not occur in Syk‐KO cells and phagosomes of Syk‐KO cells still remained surrounded by thick F‐actin structure after engulfment process. These results demonstrate that Syk accelerates phagosome acidification by facilitating the collapse of F‐actin surrounding phagosomes and resultant fusion of lysosomes to the phagosomes in complement‐mediated phagocytosis. In conclusion, our results indicate that actin‐remodeling is essential not only for phagosome formation but also for phagosome maturation process in complement‐mediated phagocytosis.

Support or Funding Information

This work was supported by JSPS KAKENHI Grant Number JP 18K07432.
Unfolded Protein Response Regulates Pulmonary P53 Expression
Authors: Akhter Mohammad Shohel , Kubra Khadeja-Tul , Uddin Mohammad Afaz , Barabutis Nektarios
Published: April 2020

INTRODUCTION

Lung endothelial barrier dysfunction leads to severe pathologies, including the lethal Acute Respiratory Distress Syndrome. P53 has been associated with anti‐inflammatory activities. The current study employs a variety of Unfolded Protein Response (UPR) activators and inhibitors to investigate the regulation of P53 by UPR in lung cells. The outcomes of our investigation reveal a positive regulation between UPR and P53. Since it has been shown that a mild induction of UPR opposes inflammation, we suggest that P53 is involved in those protective activities in the lung.

METHODS

Bovine Pulmonary Arterial Endothelial Cells (BPAEC) were maintained at 37°C in a humidified atmosphere of 5% CO2 – 21% O2, and were cultured in DMEM medium supplemented with 10% FBS and 1X penicillin/streptomycin. Protein levels were measured by Western Blot analysis, and were quantified using Image J software. All antibodies were purchased from Cell Signaling (Danvers, MA), and the chemicals from VWR (Radnor, PA).

BPAECs were purchased from Genlantis (San Diego, CA).

RESULTS

BPAEC exposed to the UPR inductors Brefeldin (2 μg/ml), Dithiothreitol (1mM), or Thapsigargin (1μM) for 4, 8 and 24 hours exerted increased expression of the UPR marker Binding Immunoglobulin Protein (BiP); as well as P53. On the other hand, treatment of those cells with the UPR suppressors N‐acetyl Cysteine (1mM), Kifunensine (5μM), or the ATP‐competitive IRE1α kinase inhibiting RNase attenuator (1μM) for the same periods (4, 8, 24 hours) exerted the opposite effects. Thus, those suppressors reduced both BiP and P53 abundance in the intracellular niche.

CONCLUSIONS

Our study suggests that UPR is directly associated with P53 expression, and that UPR induction induces P53. Since this protein supports the endothelium function, future studies may delineate the exact components of the UPR machinery in charge of that regulation, and reveal whether those effects are associated with lung endothelium integrity.

Support or Funding Information

This work was supported by 1) R&D, Research Competitiveness Subprogram of the Louisiana Board of Regents through the Board of Regents Support Fund (LEQSF(2019‐22)‐RD‐A‐26) II) NIGMS/NIH (5P20GM103424‐15, 3P20GM103424‐15S1).
MED12 Mutations Play a Pivotal Role in Breast Cancer Oncogenesis
Authors: Akula Shivani , Gonzalez Cristian , Burleson Marieke Oldenbroek
Published: April 2020

There is a critical need for superior breast cancer treatment since this disease remains the second leading cause of cancer deaths among American women. Though radiation and chemotherapy have been the frontline choice of treatment for cancer over the past few decades, due to the detrimental side effects, personalized treatment is rapidly rising as a superior treatment method. This is largely due to recent advances in genome‐wide DNA sequencing that have allowed for the identification of cancer‐related mutational landscapes. Through these technologies, MED12, a subunit of Mediator protein, has been found to be commonly mutated in a significant number of breast cancer patients. Therefore, uncovering the mechanistic basis behind MED12 mutant breast cancer is critical in order to find a personalized treatment regimen for this subclass of breast cancer. Previous work has shown that MED12 restricts GLI3‐dependent Sonic Hedgehog (SHH) signaling, a pathway that is often hyper activated in advanced breast cancer. We therefore hypothesize that mutated MED12 promotes GLI3‐dependent SHH signaling in breast cancer cells thereby promoting tumorigenesis and that this signaling pathway could be an effective therapeutic target in MED12 mutant breast cancer. To study the effect of mutant MED12, a lentivirus carrying an shRNA against MED12 was generated and infected into MCF‐7 cells. Since the vast majority of breast cancer‐associated MED12 mutations lead to loss of protein function, the knockdown strategy through lentiviral shRNA is assumed to mimic the MED12 mutant setting. Proliferation assays were utilized to confirm that downregulated MED12 plays a role in the increased proliferation of breast cancer cells. Next, quantitative PCR was performed to determine the effect of downregulated MED12 on the expression of genes that are known to be regulated by GLI3. Finally, a screening strategy was employed by using a natural compound library to find potential novel treatment strategies for MED12 mutant breast cancer. Our findings confirmed that downregulation of MED12 increases proliferation of breast cancer cells in vitro thereby providing a strong argument that mutant MED12 would have the same effect. Through quantitative PCR it was confirmed that GLI3 target genes are upregulated in cancer cells when MED12 expression is low. Importantly, our natural compound screen identified several novel therapeutic compounds that specifically target MED12 downregulated breast cancer cells through a mechanism that involves SHH signaling. We therefore uncovered that MED12 mutations promote GLI3‐dependent SHH signaling in breast cancer and we identified potential novel therapeutic strategies for MED12 mutant breast cancer patients.
The Marine Pharmacology and Pharmaceuticals Pipeline in 2020
Authors: Le Henry M , Newman David J , Glaser Keith B , Mayer Alejandro M
Published: April 2020

The ocean’s organisms continue to deliver a chemical biodiversity that contributes to both the global preclinical and clinical pharmaceutical pipelines. Thus, in November 2019, the clinical marine pharmaceutical pipeline consisted of 9 approved marine‐derived drugs: for Herpes Simplex Virus, vidarabine (Vira‐A®, FDA‐approved 1976); for pain, ziconotide (Prialt®, FDA‐approved 2004); for hypertriglyceridemia, omega‐3‐acid ethyl esters (Lovaza®, FDA‐approved 2004); and for cancer, cytarabine (Cytosar‐U®, Depocyt®, FDA‐approved 1969);, eribulin mesylate (Halaven®, FDA‐approved 2010), brentuximab vedotin (Adcetris®, FDA‐approved 2011), trabectedin (Yondelis®, EU approved 2008, FDA‐approved 2015), plitidepsin (Aplidine®) approved in Australia in 2018 and polatuzumab vedotin (POLIVY®, FDA‐approved 2019). The website www.clinicaltrials.gov was researched for marine‐derived compounds in Phase I, Phase II and Phase III: Our research determined that there were 31 marine‐derived compounds in active clinical trials: 5 marine‐derived compounds were in Phase III, 13 compounds in Phase II, and at least 9 compounds in Phase I, several auristatin‐containing antibody drug conjugates or ADCs. Regular updates on the clinical marine pharmaceutical pipeline are posted on: https://www.midwestern.edu/departments/marinepharmacology.xml

Furthermore, it is important to note that the global preclinical marine pharmacology pipeline continues to generate considerable data on multiple pharmacological classes. We thus conclude that in November 2019, both the marine pharmacology preclinical and clinical pharmaceutical pipelines remained very active.

Support or Funding Information

College of Pharmacy, Vicki Sears, Pharmacology Department, College of Graduate Studies and Office of Research and Sponsored Programs, Midwestern University
Autonomic Reactivity to Acute Mental Stress Differs Between Women and Men with Post Traumatic Stress Disorder (PTSD)
Authors: Jones Toure N. , Fonkoue Ida T. , Vemulapalli Monica , Dacosta Dana , Park Jeanie
Published: April 2020

Over 7 million Americans are affected by post‐traumatic stress disorder (PTSD), a debilitating mental illness that is associated with significant cardiovascular disease risk. Women are twice as likely as men to develop PTSD after a traumatic event. A prior study from our laboratory showed that sympathetic nervous system reactivity during combat related and non‐combat related stress is heightened in PTSD; however, this study was conducted in a predominantly male cohort and did not consider the potential influence of sex on neural cardiovascular function in PTSD. The aim of the current study was to determine if women with PTSD have heightened blood pressure (BP), heart rate (HR), and sympathetic reactivity to acute mental stress than men with PTSD. We recruited 15 women and 18 men clinically diagnosed with PTSD and confirmed using the Clinician Administered PTSD Scale (CAPS) and PTSD Checklist‐Military (PCL‐M) surveys. We measured muscle sympathetic nerve activity (MSNA), continuous BP, HR and heart rate variability (HRV) at baseline and during 3 minutes of mental arithmetic. Women and men were matched for race, age (41±2 vs 38±2 years, p=0.191), body mass index (31±2 vs 29±2 kg/m2, p=0.368), CAPS (71 ±9 vs 74 ±4, p=0.732) and PCLM (57 ±5 vs 68 ±4, p=0.129) scores. Likewise, baseline systolic BP (131±5 vs 128±4 mmHg), diastolic BP (76±4 vs 74±2 mmHg) and HR (70±3 vs 64±2 beats/min) were comparable (P>0.05) between women and men. Although resting HRV measures were not different between groups (p>0.05), resting MSNA was higher (p=0.008) in women compared to men (27±3 vs 16±3 bursts/min). In response to mental arithmetic, systolic BP reactivity was higher in women compared to men (Δ+5.1±1 versus Δ+2.2±1 mm Hg, p<0.001). Diastolic BP (p=0.366) and HR (p=0.170) responses were not different. However, MSNA reactivity was higher (p=0.04) in women compared to men (Δ15.2±5 versus Δ6.4±3 bursts/min), suggesting an exaggerated sympathetic response to acute stress. Although RMSSD reactivity was similar between both groups (p=0.347), HF response to mental stress was blunted (p=0.05) in women compared to men, suggesting a greater withdrawal of parasympathetic control of the heart during acute stress. In summary, women with PTSD show an exaggerated blood pressure and sympathetic response to acute mental stress coupled with a blunted parasympathetic control of the heart. These results provide important insight into the physiological mechanisms linking the high risk of PTSD in women and the associated cardiovascular disease and hypertension risk.

Support or Funding Information

This work was supported by National Institutes of Health (NIH) Grant R01 HL135183; NIH R61 AT010457, VA Merit I01CX001065, the Department of Veterans Affairs, Veterans Health Administration, Office of Research and Development, Decatur, Georgia, and the APS STRIDE Undergraduate Summer Research Fellowship, grant program number NHLBI; 1 R25 HL115473‐01.
Elucidating the Open and Active Conformation of Human 5‐Lipoxygenase
Authors: Gallegos Eden McMillin , Gilbert Nathaniel C. , Newcomer Marcia E.
Published: April 2020

Human 5‐Lipoxygenase (5‐LOX) is an enzyme that responds to the call for inflammation by initiating the synthesis of leukotrienes involved in asthmatic and allergic reactions and acts to sustain inflammatory reactions. Acting at the nuclear membrane in leukocytes, 5‐LOX oxygenates its lipid substrate arachidonic acid to leukotriene A4. The structure of 5‐LOX was solved in a closed conformation with helix‐α2 shortened and two large hydrophobic residues from the helix corking the active site. Most other animal lipoxygenase structures were solved with an elongated helix‐α2 allowing for an open conformation of the active site. Our goal is to study the molecular determinants important for the transition from a closed to open structure. We used site‐directed mutagenesis of helix‐α2 residues to favor either a more opened or closed conformation. We also used inhibitors to probe the conformational flexibility of 5‐LOX by testing susceptibility to cleavage by limited proteolysis and changes in melting temperature by differential scanning fluorimetry. So far, we have observed that by mutating key residues to encourage a closed conformation, 5‐LOX was less susceptible to proteolysis and melted at a higher temperature. We also showed that by mutating the same residues to encourage an open conformation, 5‐LOX was more susceptible to proteolysis. When bound to a competitive inhibitor nordihydroguaiaretic acid (NDGA), 5‐LOX melts at significantly lower temperatures and appears more susceptible to proteolytic cleavage. However, mutations that stabilize the closed conformation reduce these NDGA‐induced effects. By enhancing the presence of a closed or open conformation by mutagenesis, our results continue to hint at the structural significance of the elongated helix‐α2 in 5‐LOX dynamics. Determination of a structure of 5‐LOX in an open conformation will enable understanding of substrate acquisition and inhibitor binding. An open conformation of 5‐LOX could aid in structure‐based drug design and would foster the development of new therapies for inflammatory disorders.
Interactive Whiteboard Technology Provides a Creative Teaching Modality for a Large Neuroanatomy Laboratory
Authors: Hassan Sherif S. , Nausheen Fauzia , Scali Frank , Ettarh Rajonur
Published: April 2020

Objective

Current best teaching practices support the usage of several complementary pedagogical resources. The evolution of modern technology allowed the development of new teaching approaches that complemented the use of cadavers in the future learning model of neuroanatomy laboratory. New teaching methods were introduced to the neuroanatomy lab that included case‐based learning incorporating different neurological lesions and students were asked to work in teams to localize the neurological lesions. The aim of the present study was to assess the impact of using the Interactive Whiteboard Technology as a teaching pedagogy in neuroanatomy laboratory to verify its effectiveness over existing traditional approaches. The overall goal was to augment team‐based learning and student engagement.

Methods

A survey study of the students’ and faculties’ evaluations regarding the use of the Interactive Whiteboard Technology were statistically analyzed. These surveys were presented to the students and faculty at the end of the neuroscience course in the medical school. Students and faculty rated their learning and teaching experiences, respectively, using questionnaires. Students’ and faculties’ positive perceptions indicated their preferences to use the Panel Display in the Neuroanatomy labs.

Results

The mean score of students’ evaluations for using the Interactive Whiteboard Technology revealed high satisfaction using this technology rather than traditional learning modalities in a laboratory setting. In a case‐based learning setting, this contemporary technology enhanced critical thinking and problem‐solving skills and emphasized previously taught concepts in a more interactive and integrated way. Neuroanatomy faculty preferred the Interactive Whiteboard Technology over conventional methods when comparing brain sections with imaging and prosected images in the lab. This technology also provided a convenient and easy method for neuroanatomy faculty to share teaching materials using the quick response (QR) code which directly connects this faculty with students’ personal devices. Summative results revealed overall teaching effectiveness.

Conclusion

Using the Interactive Whiteboard Technology in neuroanatomy laboratory sessions can help students achieve their learning outcomes in lab. Using such method may overcome limitations such as the limited time for dissection. It may also allow for improved design and content of neuroanatomy laboratory for more use of small‐group sessions to be run by a smaller number of faculty.

Support or Funding Information

Self‐funded
Neural Basis of Anatomical Knowledge: An fMRI Study of Experts and Novices
Authors: Smith Theodore
Published: April 2020

How do anatomy instructors neurally organize anatomical knowledge? How does it compare to their undergraduate students’ organization? Using fMRI, we examined the neural activation patterns of expert anatomists and undergraduate anatomy students while completing an anatomy semantic task. In this talk, the outcomes of this study will be discussed then be examined in the context of curriculum and anatomical education.

Support or Funding Information

None.
KINETIC STUDIES OF NEISSERIA MENINGITIDIS SEROGROUP W CAPSULE POLYMERASE USING BIOLUMINESCENCE‐BASED ASSAYS
Authors: Moghaddam Laleh Sheikhi , McCarthy Pumtiwitt
Published: April 2020

Neisseria meningitidis is a Gram‐negative bacterium that causes meningitis. Our overall goal is increased understanding of the N. meningitidis serogroup W capsule polymerase. This enzyme creates the capsular polysaccharide found in this serogroup. During the reaction, the enzyme transfers galactose and sialic acid from two nucleotide donor sugars (UDP‐Galactose and CMP‐Sialic Acid) to an acceptor. This capsule polymerase can be used as a new tool for glycoconjugate vaccine development. In this work, we describe our efforts to determine kinetic parameters of the enzyme which are currently unknown. We have optimized commercially available bioluminescence‐based assays to investigate activity of the enzyme (CMP‐Glo and UDP‐Glo by Promega). Reactions were performed to determine Km and Vmax values in a 10‐minute reaction. Km and Vmax values of 629.2 μg/mL and 0.8965 μM/min were determined for the acceptor (hydrolyzed serogroup W polysaccharide). To simplify our studies, we have recently moved to use of a homogeneous acceptor, a sialic acid trimer (DP3) and focused strictly on the UDP‐Glo assay. In our studies with this acceptor, we confirm an increase in product formation with increasing enzyme amounts (0–1250 ng enzyme). Km and Vmax values were determined for UDP‐Galactose (44.61 μM and 0.009457 μM/min) and DP3 acceptor (2984.2 μM and 0.01099 μM/min). In future work, we will perform kinetic studies with a modified acceptor (DP3‐Galactose) to determine Km and Vmax for this acceptor and CMP Sialic acid.

Support or Funding Information

NIH 5SC2GM125517‐02NIH UL1GM118973
Sex Differences in the Cerebral Hemodynamic Response to Hypercapnia in Young Adults
Authors: Miller Kathleen B , Howery Anna J , Rivera-Rivera Leonardo A , Wieben Oliver , Barnes Jill N
Published: April 2020

Despite sexual dimorphism present in stroke and risk for Alzheimer’s disease, sex differences in cerebral blood flow regulation remain controversial. Discrepancies in the literature may be due to menstrual cycle control as well as the inherent difficulty of quantifying cerebral blood flow in humans. The purpose of this study was to evaluate the cerebral hemodynamic response to hypercapnia in young men and premenopausal women using two approaches: 1) transcranial Doppler ultrasound (TCD); and 2) 4D flow MRI. Twenty habitually physically active participants including young men (YM; n = 10, age = 24±2 y) and young women (YW; n = 10, age = 26±4 y) were studied. Young women were studied during days 2–6 of the menstrual cycle. During separate, randomized study visits, left middle cerebral artery (LMCA) velocity was measured with TCD and LMCA flow was measured with 4D flow MRI during normocapnia and two levels of hypercapnia (4% and 6% CO2). 4D flow MRI quantifies cerebral hemodynamic and angiographic data simultaneously. End‐tidal carbon dioxide and mean arterial pressure were continuously monitored with an anesthesia monitor. At baseline, LMCA velocity (YM: 53±3 cm/s vs. YW: 60±4 cm/s, p=0.15) and LMCA flow (YM: 121±10 ml/min vs. YW: 131±6 ml/min, p=0.41) were not significantly different between groups. There was a trend for YM to demonstrate a larger LMCA cross‐sectional area compared with YW at baseline (YM: 0.071±0.002 cm2 vs. YW: 0.065±0.002 cm2, p=0.052). In response to hypercapnia, the LMCA velocity response was not different between groups (YM: 1.17±0.29 cm/s/mmHg vs. YW: 1.24±0.17 cm/s/mmHg, p=0.83). However, YM demonstrated a greater LMCA flow response compared with YW (YM: 2.98±0.40 ml/min/mmHg vs. YW: 1.38±0.26 ml/min/mmHg, p<0.01), even when accounting for changes in blood pressure. In addition, the LMCA significantly vasodilated in YM in response to hypercapnia (p<0.05); however, there was no significant change in LMCA cross‐sectional area in response to hypercapnia in YW (p=0.13). These results suggest that young men have augmented cerebral blood flow response to hypercapnia compared with young women. In addition, due to small changes in vessel cross‐sectional area during hypercapnia, sex differences may be underestimated if cerebral blood velocity is used as a surrogate of blood flow.

Support or Funding Information

Supported by the Wisconsin Alumni Research Foundation and the National Institutes of Health T32 HL‐007936.
Anatomical basis of the temporal muscle trigger points
Authors: Akamatsu Flávia Emi , Teixeira Giulianna Simonetti , Itezerote Ana Maria , Hojaij Flávio , Andrade Mauro , Jacomo Alfredo Luiz
Published: April 2020

Background

Myofascial syndrome is a common cause of chronic musculoskeletal pain associated with activity restrictions and is characterized by myofascial trigger points (MTP). MTP are clinically identified by palpation of a muscle or fascial taut band. Pathophysiology of MTP is unclear, coincidence between them and muscle motor plate has been suggested. The temporal muscle is related to headache and temporomandibular disorders. Pain radiates from hyperalgesic points to distant areas just as it is reported in patients with neck pain and symptoms in the orofacial region. Temporal muscle locates trigger areas and trigger points in various parts of the muscle. Four trigger points are selected, but an anatomical correlation is missing

Methods

Temporal muscles from six cadavers were carefully dissected from their origins in order to observe the exact point where nerve fibers penetrated the muscle belly. As muscle size varies among individuals, we calculated the relative entry point of the nerve into the muscle by defining six different areas in the muscle belly: three superior (I, II and III) and three inferior (IV, V and VI), beginning at the posterior margin of the muscle. Statistical analyzis OF anatomical data was obtainded by Poisson distribution and logarithm link function followed by Bonferroni multiple comparisons. (p <0.05)

Results

Areas II and III had the largest number of temporal nerve entry sites, with a mean of 3 sites (25%), followed by quadrant V and VI with a mean of 2 sites (16%) and quadrant I and IV with a mean of 1 site (9%).

Conclusion

In accordance with the clinical literature, the branches of the temporal nerve in the temporal muscle corresponded to the described areas of the myofascial trigger points (MTPs). Anatomical correlations between MTPs may be a useful tool for a better understanding of the physiopathology of these disorders and may provide a rational basis for their treatment.
Temporal behavior of weight loss in a rat model of gastric restriction
Authors: Akamatsu Flávia Emi , Itezerote Ana Maria , Saleh Samir Omar , Hojaij Flávio , Andrade Mauro , Martinez Carlos , Jacomo Alfredo Luiz
Published: April 2020

Background

Obesity is a complex and multifactorial disorder and has become a major healthcare burden in some countries. The rising prevalence of overweight and obesity is considered a global pandemic. The current paradigm in obesity treatment has traditionally relied upon lifestyle modifications (diet and exercise) and pharmacological treatments which often lead to modest results. Surgical procedures have been increasingly performed. Bariatric surgery results in superior outcomes in terms of weight loss and weight‐associated comorbidities as compared to non‐surgical interventions. Experiments with animal models have significantly helped to elucidate some of the potential mechanisms underlying bariatric surgery. There is no information about gastric restriction with phytobezoar in animal model and the weight loss.

Purpose

Measure the weight loss until 20 days after surgical gastrostomy and phytobezoar insertion and compare to sham and normal group

Methods

Experimental model of gastric restriction was devised using rats. The animals were submitted to surgical gastrostomy and a cylindrical loofah was inserted into the stomach. We studied 30 adult male Wistar rats divided in three groups: the stomach reduction group (R20), the sham group (SH20), which underwent the same procedure except for the loofah insertion and the control group (C20). Animals were fed and kept in separate cages. They were weighed every 3 days until being sacrificed on day 20. The results were described by the mean value with respective standard deviation. Comparison of multiple groups was performed using analysis of variance (ANOVA) followed by Bonferroni post hoc tests. All tests were applied with 5% significance level.

Results

There was a mean difference in the behavior of weight changes over the evaluation days between the groups (p Interaction <0.001), and on all evaluated days the experiment group presented a lower average weight change than the other groups (p <0.001). These findings might be a result of post‐ surgical weight loss correlated to a large extent with reduced spontaneous eating. This indicates that in our animal models it seems to recapitulate remarkably well the findings in humans.
Mitigating Effect of Korea Red Ginseng Against Multi‐Tissue Iipotoxicity Caused by High‐Fat Diet Feeding
Authors: Kim Sung Hyeok , So Gyeongseop , Ha Chang Woo , Park Yuna , Lee Jin Woo , Jang Seon-A , Sohn Eun-Hwa
Published: April 2020

Backgrounds

Excessive consumption of fat and its aberrant deposit in the body has been linked to the low‐grade inflammation in various susceptible organs and eventually causes tissue toxicity. This study examines the multifaceted suppressive effects of Korean Red ginseng extract (KRG) against high‐fat diet (HFD)‐induced lipotoxicity and inflammatory responses occurring in the liver, aorta, and brain.

Methods

The standardized KRG powder was prepared by Korea Ginseng Corporation and was dissolved in phosphate buffered saline as a concentration of 30 mg/mL. Male C57BL/6 mice were fed HFD with or without KRG (10 or 50 mg/kg body weight; orally) for 12 weeks (n = 10 per group). The anti‐inflammatory potential of KRG was determined in the blood and the aorta, liver, and brain.

Results

KRG significantly inhibited HMG‐CoA reductase activity in vitro. In vivo, KRG supplementation suppressed HFD‐associated body weight gain, lipid profile changes, excessive fat deposition in the liver, and increases of leptin, insulin, and ALT levels in the blood. Inflammatory markers of the aorta (ICAM‐1 protein level and wall thickening of the aorta), liver (COX‐2 expression and liver function tests), and brain (APP, BACE1, and Tau protein levels) were also significantly reduced by KRG. In microvascular endothelial cells, 15% cyclic stretch‐mediated upregulation of ICAM‐1 and VCAM‐1 expression was significantly attenuated in the presence of KRG.

Conclusion

KRG supplementation attenuates HFD‐mediated body weight gain, lipid profile changes, and multi‐tissue inflammatory responses including the liver, aorta, and brain.