bluuebunny/crossref_metadata_2025_split huggingface dataset:
Cardiac Gene Expression and Histology in a Rat Model of Fat Embolism
Authors: Wacker Michael J. , Patel Shaan , Vallejo Julian , Colson Jordan , Edegbe Joy , Lin Jane , Salzman Gary , Poisner Alan , Monaghan A. Paula , Molteni Agostino 
Published: April 2020

IntroductionFat Embolism consequent to Triolein (T) intravenous injection in rats induces severe histopathologic pulmonary damage at 48 hours mostly related to vasculitis, peribronchial and septal inflammation with an increase in mast cells, and eventually, organ fibrosis. We previously reported that the hearts of the same animals at 48 hours did not show vasculitis of coronary arteries branches, inflammation or an increase in mast cells. The objective of this study was to determine if T treatment by 10 weeks induced changes in cardiac gene expression or histopathology related to fat embolism.Materials and Methods24 Sprague Dawley Rats (250–330g) were divided into two groups, 48 hour treatment and 10 week treatment (N=12 each). Six rats of each group were intravenously injected with 0.2 ml of T, and the other 6, 0.2 ml of normal saline. The rats were euthanized under general anesthesia 48 hours or 10 weeks after injection. A cut was made in hearts along the transverse plane; the top half of hearts were snap frozen in liquid nitrogen for gene expression studies and the bottom half was fixed in formaldehyde for histopathological evaluation similar to lung preparations. After RNA isolation, real time RT‐PCR was performed on genes associated with pathological cardiac hypertrophy or fibrosis (beta myosin heavy chain, alpha myosin heavy chain, skeletal muscle alpha actin, atrial natriuretic peptide, brain natriuretic peptide, vimentin, and collagen 1a1).ResultsDespite significant damage to the lungs, there was no significant changes in expression of any of these genes in the hearts of the rats treated with T, versus those of the saline treated group, whether sacrificed at 48 hours or 10 weeks post injection (P>0.05 ANOVA). Histopathological evaluation of the hearts at 10 weeks also did not show any damage in the myocardium in sections stained by H and E and no vasculitis, mast cell increase, or ventricular enlargements. Trichrome staining did not show any fibrotic increase in hearts of T injected animals, thus confirming the previous observations and gene expression analysis.ConclusionThere was a marked contrast in organ damage between hearts and lungs in this study in rats with T treatment. Hearts demonstrated no significant change in gene expression or histology demonstrating a lack of fibroblast infiltration, hypertrophy, or remodeling due to T treatment. Kidney, liver, brains, and aortas also showed varying degrees of histopathological damage compared to the severity of the damage of the lungs. The renin angiotensin system plays a key role in the pathogenesis of the lung damage with fat embolism and this system needs to be further explored for organ differences to fat embolism induced by T treatment.Support or Funding InformationSupported by the Catherine T. Geldmacher Foundation, St. Louis, MO
Quantitative 3D Reconstructions of the Human Superior Olive Complex are Consistent with Auditory Dysfunction in Autism Spectrum Disorder
Authors: Kulesza Randy , Mansour Yusra 
Published: April 2020

Beyond the characteristic social, communication and behavioral aberrations associated with autism spectrum disorder (ASD), the vast majority of subjects with this condition have some degree of auditory dysfunction. The auditory deficits range from complete deafness to hypersensitivity but typically include difficulty listening to speech in the presence of background noise. Clinical testing of brainstem evoked potentials and stapedial reflexes in children and adults with ASD reveal lower thresholds but slower conduction times and right‐left ear asymmetries. Previous studies on post‐mortem brain specimens from our lab revealed drastic and consistent hypoplasia and dysmorphology in the auditory brainstem. The most drastic alterations were found in the medial superior olive (MSO), the largest and most prominent component of the superior olivary complex (SOC). Our morphological studies have shown that in subjects with ASD, there are significantly fewer neurons in the MSO and surviving neurons are smaller, more round and have an abnormal orientation. Based on these results, we hypothesized that the SOC in subjects with ASD not only includes fewer neurons but that these nuclei occupy significantly less brain volume and demonstrate abnormal nuclear contours. We investigated this hypothesis by obtaining donated brain tissue from an age‐matched series of subjects with ASD or no known neurological diagnosis (neurotypical; NT) through Autism BrainNet. Free‐floating tissue sections were mounted onto glass slides, stained for Nissl substance with Giemsa and sealed under coverslips. 3D volume renderings of the SOC nuclei were created in Amira. Subjects in this study include two NT subjects (ages 4 and 8, both female) and four age‐matched subjects with ASD (ages 2–11 years of age, all male). At autopsy, there was no difference in total brain weight between NT and ASD subjects (p=.42). Our 3D reconstructions reveal a drastic reduction in the total volume of the SOC in subjects with ASD (control = 6.8 mm3, ASD = 2.23 mm3; p<.005). In fact, all SOC nuclei were significantly smaller in ASD subjects expect for the ventral nucleus of the trapezoid body (p=.922). Together, these results support the brainstem as a core site of pathology in ASD and are consistent with previous clinical evaluations of hearing in subjects with ASD.Support or Funding InformationAutism BrainNet
Lipocalin 2 Induces Neuroinflammation through Liver‐Brain Axis in Nonalcoholic Fatty Liver Disease
Authors: MONDAL AYAN , SETH RATANESH , SARKAR SUTAPA , BOSE DIPRO , SAHA PUNNAG , KIMONO DIANA , ALBADRANI MUAYAD , NAGARKATTI MITZI , NAGARKATTI PRAKASH , CHATTERJEE SAURABH 
Published: April 2020

Nonalcoholic fatty liver disease (NAFLD) is chronic liver disorder characterized by accumulation of excess fat in liver due to consumption of high fat/lipid diets. A multiple hit paradigm that include lipotoxicity, environmental and genetic causes for prolonged period often leads to nonalcoholic steatohepatitis (NASH) with scarring and inflammation in liver that consequently causes irreversible hepatic damage. Both clinical and basic research implicated a strong correlation between NAFLD/NASH phenotypes with several ectopic manifestations like type 2 diabetes, cardiovascular disease, chronic kidney disease and more recently neuroinflammation and neurodegeneration. Notably, mechanisms of neuroinflammation in underlying NAFLD are poorly understood. In this report we identified adipokine Lipocalin 2 (Lcn2) induced neuroinflammation by acting through liver‐brain axis in diet induced murine NAFLD/NASH model. We found that Lcn2 is overexpressed in liver and in portal circulation that activated 24p3R (Lipocalin2 receptor) in brain and induced the release of high mobility group box 1 (HMGB1) from brain cells. Released HMGB1 acted as a preferential ligand to Toll like receptor 4 (TLR 4) and induced oxidative stress by activation of NOX‐2 signaling. NOX2 downstream signaling activated NF‐kB by phosphorylation of p65 protein. Activation of p65 and NOX‐2 led to NLRP‐3 inflammasome activation which subsequently released proinflammatory cytokines IL‐6 and IL‐1β from brain cells. Furthermore, we show that elevated Lcn2 level from neurons in underlying NAFLD caused a non disruptive blood brain barrier dysfunction by over production of TNFα, IL‐1β and MMP 9 without alteration of tight junction proteins Claudin 5 and Occludin. Based on the above, the above findings identify Lcn2 as a potent molecule inducing neuroinflammation in underlying NAFLD.Support or Funding InformationNIH#P01ES028942‐01 to Dr. Saurabh Chatterjee
Nocodazole‐induced growth inhibition of human leukemia cells is linked to downregulation of CAK molecules
Authors: Almeida Adrian , Baquier Jocelyn , Hu Xiaotang 
Published: April 2020

A cytoskeleton is present in the cytoplasm of all living cells. The cytoskeleton is composed of three different types of fibers: microtubules, microfilaments, and intermediate. The microtubules play an important role in a variety of cellular processes including maintaining cell structure and pulling chromosomes apart during anaphase. Nocodazole is an inhibitor of microtubule polymerization. Inhibition of microtubule polymerization by Nocodazole often lead to arrest of cells in G2 or M phase. However, it is unclear if any cell cycle regulatory molecules are involved in Nocodazole‐mediated cell growth. In this study we focus on CDK activating kinase (CAK). The CAK is composed of three subunits, CDK7, cyclin H, and MAT1. The cyclin H is regulatory subunit of CDK7 whereas MAT1 determines the specificity of CDK7. Addition of Nocodazole significantly inhibited the growth of MV4‐11 and TF‐1a cells in culture. The inhibitions were dose‐dependent with maximal inhibition being observed at a concentration of 10uM of Nocodazole. Less than 1 μM had no significant inhibitory effect on the growth of the cells, measured by XTT proliferation assay. Paralleled to the changes in the cell proliferation, expression of CDK7 and cyclin H were marked decreased when Nocodazole were added to TF‐1a and MV4‐11 cells at concentrations of 5 and 10μM. Similarly, Nocodazole at concentration of 10μM downregulated the phosphorylation levels of CDK7 and the expression of RNA polymerase II in TF‐1a cells. However, Nocodazole had no effect on the expression of MAT1, suggesting that Nocodazole did not interfere with specificity of CDK7. We also found that Nocodazole had no effect on the expression of β‐actin, a non‐muscle cytoskeleton actin.Support or Funding InformationBarry University
Infarct macrophage secretome coordinates neutrophil degranulation
Authors: Daseke Michael , Kalusche Will , Flynn Elizabeth , Konfrst Shelby , Lindsey Merry 
Published: April 2020

Following myocardial infarction (MI), the left ventricle undergoes wound healing that begins with an intense inflammatory response to recruit leukocytes to the infarcted region. Infiltrated neutrophils degranulate, releasing a series of proteases including matrix metalloproteinase (MMP)‐9 into the extracellular space. Macrophages also infiltrate into the infarct region, and we hypothesized that the macrophage may secrete factors that regulate neutrophil degranulation. Stimulating bone marrow derived neutrophils from C57BL/6J mice (3–6 month old male) with phorbol myristate acetate (PMA) induced degranulation, as evidenced by release of MMP‐9. Co‐stimulation with MI day 1 macrophage secretome (10% by volume) reduced PMA‐induced degranulation of MMP‐9 by 8‐fold (p<0.0014). Transcriptomics analysis of day 1 MI macrophages revealed galectin‐3, vimentin, fibronectin, and murinoglobulin‐1 were highly overexpressed. Examination of the day 1 MI macrophage secretome also showed high expression of vimentin and fibronectin. To further unmask signaling connections, neutrophils were co‐stimulated with PMA and day 1 MI macrophage secretome, along with blocking antibodies for the 4 proteins individually. Galectin‐3 or fibronectin inhibition promoted while murinoglobulin‐1 inhibition reversed the macrophage secretome effect. Neutrophil stimulation with recombinant galectin‐3 or fibronectin showed an additive effect with PMA on MMP‐9 release. Vimentin inhibition or stimulation had no effect on neutrophil degranulation. Overall, our results uncovered a role for galectin‐3 and fibronectin in inducing neutrophil degranulation while murinoglobulin‐1 may be a potent inhibitor of degranulation. The day 1 MI macrophage, therefore, secretes both factors that promote as well as factors that temper neutrophil degranulation. The balance of these factors determines the net effect of the macrophage secretome on neutrophil degranulation.Support or Funding InformationNIH HL075360, HL129823, HL13731, VA 5I01BX000505.
Bacterial cell wall modification by glycolipid probes
Authors: Marando Victoria M. , Calabretta Phillip J. , Hodges Heather L. , Kraft Matthew B. , Kim Daria E. , Kiessling Laura L. 
Published: April 2020

Despite the ubiquity and importance of glycans in biology, methods to probe their structures in cells are limited. The structures of mammalian glycans can be perturbed through metabolic incorporation, in which non‐natural sugars are taken up by cells, converted to nucleotide‐sugar intermediates, and incorporated via biosynthetic pathways. Applications of metabolic incorporation in bacteria are complicated by the diversity of pathways bacteria use to acquire, metabolize, and catabolize monosaccharide intermediates. We developed a strategy to alter glycan structure that avoids intracellular processing using lipid‐linked glycans, intermediates directly used in glycan biosynthesis. We generated synthetic arabinofuranosyl phospholipids to test this strategy in Corynebacterium glutamicum and Mycobacterium smegmatis, organisms that serve as models of Mycobacterium tuberculosis. Using a C. glutamicum mutant that lacks arabinan, we identified glycolipid donors whose addition restores cell wall arabinan, demonstrating that non‐natural glycolipids can serve as biosynthetic intermediates and function in chemical complementation. Our lipid‐based probe could rescue wild type cells treated with an inhibitor of cell wall biosynthesis. We then focused on incorporating derivatives with chemical handles for labeling such that bacterial glycans can be captured and visualized. To this end, we have generated the first probe to label arabinose‐containing structures in mycobacteria and corynebacteria.Support or Funding InformationNIH: RO1‐AI063596, NSERC PGS‐D
Anti‐inflammatory effects of vagus nerve simulation in a newborn rat model of acute inflammation
Authors: Cacho Bradley , Abdala Jonathan , Wilson Christoper G. 
Published: April 2020

Neonatal sepsis and inflammation pose a potential source of morbidity and mortality in newborns, specifically in the first weeks of life. We have previously shown that vagus nerve stimulation modulates pro‐inflammatory cytokine expression in the Central Nervous System (CNS). The optimal vagus nerve stimulation (VNS) parameters to reduce inflammation have been unknown.In this study, we tested the hypothesis that vagus nerve stimulation reduces the expression of pro‐inflammatory cytokines in a lipopolysaccharide (LPS) model of airway inflammation. To test our hypothesis, we used intratracheal injections of LPS (0.5 mg/kg in 10 μL of saline) in neonatal Sprague Dawley rats (postnatal days 10 to 12) to induce pro‐inflammatory cytokine production. The P10–12 pups were anesthetized with isoflurane (2 to 4%) and then LPS was injected into the trachea to mimic airway inflammation. The experimental group was subjected to VNS at pre‐selected frequencies (ranging logarithmically from 10 to 10000 Hz) continuously for 30 minutes immediately after the LPS injection. Stimulation current was titrated down as needed to account for irregular breathing or apneas (typically approximately 0.5 mA). After recovery, each rat’s brainstem was collected and processed for cryosectioning and immunohistochemistry. We stained for and measured the expression of IL‐1 beta, TNF alpha, and HMGB1 in subjects with or without VNS (the positive control group received LPS without VNS). We then used ImageJ software and the particle analysis plug‐in to quantify the number of cells that expressed cytokines in the nucleus tractus solitarius (NTS).Using one‐way ANOVA analysis, our data showed a significant decrease (p < 0.05) in expression of IL‐1 beta, TNF alpha, and HMGB1 in LPS‐injected rats that underwent VNS compared to LPS‐injected rats that did not receive VNS. Frequencies of 10, 100, 1000, and 10000 Hz all resulted in significant decreases in cytokine expression. Cytokine expression in the LPS + VNS group was similar when compared to sham rats (no LPS injection or VNS). Our study showed that VNS significantly decreased the expression of the early inflammatory cytokines in LPS‐induced inflammation in neonatal rats. Blocking the early cytokine cascade may be associated with decreased inflammation and a concomitant reduction in associated symptoms in patients with sepsis. VNS used either alone or in conjunction with antibiotics may lead to improved outcomes in neonatal patients suspected to be affected by infection.Support or Funding InformationThis study was supported via a grant funded by the Pediatric Research Initiative at Loma Linda University.
Rice bran supplementation improves glucose tolerance and innate immunity in high‐fat diet induced obese ovariectomized mice
Authors: Chen Li Hsin , Yeh Chiu Li 
Published: April 2020

ObjectivesMenopause is a state of low estrogen production which is associated with chronic low‐grade inflammation and higher incidence of overweight. Rice bran (RB) is a nutrient‐dense product of rice milling process that is considered a health improving supplement. The aim of this study was to investigate the influences of RB on the innate immunity of high‐fat diet induce ovariectomized (OVX) obese mice.Methods8 months‐old female mice were randomly divided into OVX sham (SH) group and OVX group. After surgery for one week, mice were randomly assigned to one of the three groups (n=12) and fed the respective diet for four months : SH group and OVX‐control (ONC) group were provided with AIN‐95 control diet containing 10% energy as fat, OVX‐high fat diet (OHF) group was treated with high fat diet (HFD) containing 40% fat, mice in the OVX‐high fat diet with RB (OHFR) group were fed with HFD containing RB (10% wt/wt).ResultsMice in the OHF group showed significantly higher body weight gained, plasma glucose and triglyceride levels than those in the SH and other OVX groups. Compare to the SH and ONC groups, the OHF group had lower splenic natural killer (NK) cells cytotoxic capacity and peripheral blood phagocytes phagocytic ability. The chemotactic ability of leukocytes were also lower in the OHF group. As regard to the OVX group with high‐fat diet, RB supplementation increased NK cells cytotoxic function and phagocytosis ability of phagocytes. Also, the chemotactic ability of neutrophils and macrophage as measured by migration speed and directional persistence were improved in the OHFR group.ConclusionThe findings of this study suggest that RB supplementation improved glucose tolerance and prevented the decline of innate immunity in high‐fat diet induced obese mice with OVX.Support or Funding InformationThis study was supported by research grant MOST 107‐2320‐B‐038‐014 from the Ministry of Science and Technology (Taipei, Taiwan).
Reduction of Liver Heparan Sulfate Proteoglycans Promotes Endocrine Benefits of Therapeutic FGF1
Authors: Vi Quach Anthony , Pessentheiner Ariane , Gordts Philip 
Published: April 2020

Type‐2 diabetes, a is a global epidemic that currently drives an enormous health and economic burden. The disease is chronic condition in which blood glucose levels are deregulated due to insulin resistant in metabolic tissues, such as liver, muscle and adipose tissue. Recently, fibroblast growth factor 1 (FGF1) has been implicated in promoting insulin‐depended glucose uptake in adipose tissue. Therefore, FGF1 is being considered for therapeutic use in type‐2 diabetes patients.Typically, both FGF and FGF Receptors (FGFR) bind heparan sulfate proteoglycans (HSPGs).This multimetric FGF‐FGFR interaction with HSPGs promotes stable complex formation allowing robust intracellular signaling. Our preliminary results show that the degree of HSPG sulfation in adipose tissue is critical for FGF1 mediated glucose lowering. The liver expresses significant amounts of FGFRs and highly sulfated cell surface HSPGs. Therefore, we hypothesize that the liver is a prime scavenger of therapeutic FGF1 reducing its partitioning to FGFR in adipose tissue and consequently its therapeutic window.To address this hypothesis, we made use of wildtype and genetically modified mice with reduced liver HSPG sulfation introduced by liver specific inactivation of one of the biosynthetic enzymes N‐deacetylase and N‐sulfotransferase 1 (NDST1; Ndst1f/fAlbCre+). Both mice were fed a 60% kcal high fat diet (HFD) to initiate diet‐induced obesity and the associated peripheral insulin resistance. Throughout the HFD treatment, we monitored the mice’s glucose and body weight on a biweekly basis and found no phenotypic changes between the wildtype Ndst1f/fAlbCre+ mice. After 10 weeks on the HFD, we tested each mice group’s insulin and glucose tolerance via injections of the corresponding substrates and saw no significant differences in glucose homeostasis. By week 15 of HFD feeding, we injected a single dose of recombinant FGF1 protein (0.5 mg/kg) and recorded the FGF1‐induced glucose lowering in periodic intervals following the injection. Ndst1f/fAlbCre+ mice presented with a greater reduction in FGF1‐induced glucose lowering as well as longer lasting reduction effect.Overall, the results from this research will provide an enhanced understanding of FGF1 and HSPGs that could contribute to future studies and treatments of type‐2 diabetes and overall help increase the therapeutic window of FGF1 treatment in type‐2 diabetes.Support or Funding InformationHave been awarded Undergraduate Research Scholarship grant for working as a full‐time researcher during a 10‐week period.
Heat shock proteins in embryonic and adult submandibular salivary glands in healthy and tumorigenic tissues
Authors: Basset Charbel , Cappello Francesco , Rappa Francesca , Florena Ada Maria , Lentini Vincenzo Luca , de Macario Everly Conway , Macario Alberto J. L. , Jurjus Abdo , Jurjus Rosalyn , Leone Angelo 
Published: April 2020

IntroductionHeat Shock Proteins (HSPs) role in development and function of human salivary glands have not yet been measured or mapped in detail, in particular, HSP60 role is unknown. They can be potential biomarkers for glandular diseases and malignancies. We present our initial findings on HSP10, HSP27, HSP60, HSP70 and HSP90 distribution in human submandibular salivary glands (SMG) at various stages of development and tumorigenesis.MethodsAdult human SMG (normal and tumoral) and embryonic head tissue samples were processed by standard methods for routine histological analysis and immunohistochemical staining using antibodies against the five HSP’s. All procedures were in accordance with the Helsinki Declaration. In addition, proteins and RNAs were extracted from normal SMG of FVB‐NJ adult mice. Protein and gene expression of HSPs were assessed by western blot and RT‐PCR respectively.ResultsHSP27, HSP60 and HSP90 were present in the acini and ducts of embryonic salivary glands but had a different distribution pattern in adult glands: HSP27, HSP60 and HSP10 occurred only in the ducts whereas HSP90 and HSP70 were totally absent in the adult gland. The results were supported by our western blot and RT‐PCR as HSP10, HSP27 and HSP60 protein and mRNA were expressed in control adult mice SMG. HSP70 and HSP90 protein and mRNA were not expressed in adult mice SMG, thus matching our immunohistochemical results. In contrast, all 5 HSP’s levels were high in Pleomorphic Adenoma, Warthin’s tumor and Adenoid Cystic Adenoma.ConclusionsThe HSP27, HSP60 and HSP90 participate actively in the developmental process. HSP’s are also involved in neoplasm formation and progression. These results support the diagnostic and prognostic potentials of HSP’s.SignificanceSalivary glands tumors incidence is increasing, which makes it necessary to develop diagnostic tools and biometrics to understand the molecular mechanisms involved. Since molecular chaperones show quantitative variations and contribute to the carcinogenic mechanism in a variety of cancers, we aimed to investigate if a similar situation occurs in salivary glands and characterize the functions of the chaperoning system in physiology and pathology in these glands. Our findings exhibited quantitative patterns of the HSP’s, which can distinguish between normal and tumor tissues of the SMG and have potential in diagnosis and patient monitoring.