bluuebunny/crossref_metadata_2025_split huggingface dataset:
Genotype‐Phenotype Correlations for Protein O‐Linked Mannose NAcetylglucosaminyltransferase 1 in Congenital Muscular Dystrophy
Authors: Ho Jessica Ziling , Boyd Sally Riewe , Singh Danish , Halmo Stephanie Marie , Wells Lance 
Published: June 2021

Congenital muscular dystrophy (CMD) is a heterogeneous family of inherited muscle disorders. A subtype of CMD known as dystroglycanopathy is classified by hypoglycosylation of alpha‐dystroglycan (α‐DG). α‐DG, an integral component of the dystrophin‐glycoprotein complex, is responsible for connecting the actin cytoskeleton to the extracellular matrix in muscle tissues. Hypoglycosylation of α‐DG arises from defects in the protein O‐mannosylation biosynthetic pathway. One enzyme involved in the O‐mannosylation pathway is protein O‐linked‐mannose β‐1,2‐N‐acetylglucosaminyltransferase (POMGNT1). On serine and threonine residues in α‐DG, POMGNT1 catalyzes addition of N‐acetyl‐D‐glucosamine (GlcNAc) to an O‐mannose structure in a β1,2‐linkage to create a core m1 glycan structure in the cis‐Golgi. Mutations in the gene encoding POMGNT1 have been observed in patients with various forms of dystroglycanopathy.Our work examines the role of R311Q, P493R, T176P, P303L, C269Y, E223K, C490Y, G502A, R605P, I287S, E156K, D395N, and D556N mutations in POMGNT1 to define a genotype‐phenotype correlation. The POMGNT1 mutations present a range of clinical presentations, from muscle‐eye‐brain disease, an extremely severe condition characterized by muscle weakness, vision and brain abnormalities, and developmental disabilities, to limb‐girdle muscular dystrophy, a much milder phenotype that results in progressive muscle weakness and wasting but lacks neurological comorbidities. The effects of the selected mutations on enzyme characteristics are not well established in the literature. Therefore, we sought to identify mutation‐derived changes in enzyme kinetics and stability.To do so, HEK293F cells were transfected with mutant plasmids generated by site‐directed mutagenesis. Preliminary data indicated that R311Q, D395N, R605P, and D556N POMGNT1 mutants maintained enzyme expression. Radiolabel transfer assays established that the R311Q, D395N, and R605P mutants were kinetically dead while the D556N mutant still exhibited transfer. Promega's UDP‐Glo™ assay was performed on the D556N mutant, which demonstrated reduced kinetic activity compared to wild‐type POMGNT1. A SYPRO Orange thermal shift assay revealed the aforementioned POMGNT1 mutants to be thermodynamically stable. We are currently experimenting with the rest of the mutants and are also investigating the ability of all mutants to rescue POMGNT1 knockout cell lines for IIH6 reactivity, laminin binding, and Lassa pseudovirus entry. Understanding genotype‐phenotype correlations in those glycosyltransferases will facilitate the design of more targeted treatments for individuals based on the mutation(s) they carry.Support or Funding InformationThis work was supported in part by NIGMS/NIH (R01GM111939, LW).This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
MMP‐12 knockout mouse: Is it a relevant animal model to study the role of MMP‐12 in post‐stroke brain damage?
Authors: Nalamolu Koteswara Rao , Chelluboina Bharath , Magruder Ian B. , Mohandas Adithya , Venkatesh Ishwarya , Klopfenstein Jeffrey D. , Pinson David M. , Wang David Z. , Veeravalli Krishna Kumar 
Published: June 2021

We recently reported for the first time the detrimental role of matrix metalloproteinase (MMP)‐12 on post‐stroke brain damage. We hypothesize that the post‐stroke MMPs dysregulation is similar across species and the genetic deletion of MMP‐12 does not affect the post‐stroke expression of other MMPs. We tested our hypothesis by determining the pre‐ and post‐ischemic expression profile of MMPs in MMP‐12 knockout and respective wild type mice. Focal cerebral ischemia was induced in MMP‐12 knockout and wild type mice by middle cerebral artery occlusion procedure by insertion of a monofilament suture. One hour after ischemia, reperfusion was initiated by removing the monofilament. One‐day after reperfusion, ischemic brain tissues from various groups of mice were collected for real‐time PCR analysis and PCR analysis followed by agarose gel electrophoresis. Although the post‐stroke regulation of several MMPs was altered in wild type mice, huge upregulation of MMP‐12 (~763 fold over sham) was noticed in mice similar to rats as reported earlier by our group. Unlike in rats, MMP‐8 was prominently upregulated (~280 fold over sham) in ischemic mice brains. As expected, MMP‐12 expression is absent in the ischemic brains of MMP‐12 knockout mice. Except the expression of MMP‐9, the expression of several other MMPs such as MMP‐1a, −2, −8, −10, −15, −16, −17, −21 and −28 was altered in the ischemic brain of MMP‐12 knockout mice as compared to wild type mice under similar experimental conditions. In addition, the expression of various MMPs in MMP‐12 knockout mice under non‐ischemic conditions was altered as compared to the wild type mice that did not subject to ischemia. Based on our results, we conclude that the mice with genetic deletion of MMP‐12 are constitutive in nature that warrants the attention of their use in elucidating the role of MMP‐12 in post‐stroke brain damage.Support or Funding InformationThis work was supported in part by the OSF HealthCare Foundation and OSF Illinois Neurological Institute, Peoria, IL.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Role of microRNA‐9 in the pathogenesis of Parkinson's Disease
Authors: Goh Suh Yee , Yeo Xin Yi , Srinivasan Dinesh Kumar , Dheen S Thameem , Tay Samuel Sam Wah 
Published: June 2021

Parkinson's disease (PD) is the second most prevalent neurodegenerative disease in the aging population. Previously, our lab has shown that miR‐9 is upregulated in the substantia nigra pars compacta (SNc) in pre‐clinical models of PD [1‐methyl‐4‐phenyl‐1,2,3,6‐tetrahydropyridine (MPTP)‐treated mice]. Briggs et al (2015) has also reported this trend in the SNc of post‐mortem PD patients. miRNAs are post‐transcriptional regulators of gene expression that are involved in the maintenance of normal cellular functions. While miR‐9 is well‐studied as a regulator of neurogenesis during brain development, its upregulation observed in PD suggests that miR‐9 may have other functions that contributes to PD pathogenesis. In this study, we are interested to investigate the miR‐9‐mediated mechanisms involved in the development of PD. Using miRNA qPCR, we verified the increase in miR‐9 levels in in vitro PD models [MN9D and SH‐SY5Y neuronal cells treated with 1‐methyl‐4‐phenylpyridinium (MPP) iodide]. MPP treatment decreased neuronal cell viability by about 30% and the inhibition of miR‐9 significantly reversed this trend, suggesting that miR‐9 plays a role in neurodegeneration. In addition, using NanoSight technology, we discovered a significant increase in the concentration and size of exosomes released by neuronal cells treated with MPP iodide. Interestingly, these exosomes also contained higher miR‐9 levels as compared to exosomes released by neurons under control conditions. This suggests that miR‐9 may be transported to other cell types (such as microglia) to regulate other events in PD, for instance, neuroinflammation. These research findings implicate miR‐9 as a potential diagnostic biomarker as well as a potential therapeutic target for PD treatment.Support or Funding InformationThis work was supported by a research grant from the Singapore Ministry of Education Academic Research Fund Tier 1.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
A Comparison of Multiple Histotechnical and Morphological Methods for Measuring Hepatic Lipids in NASH Models
Authors: Opsahl Alan C. , Crowell Danielle M. , Zhan Yutian , Geoly Frank J. , O'Neil Shawn P. 
Published: June 2021

Nonalcoholic steatohepatitis (NASH) is a progressive syndrome in which fat (usually in the form of triglycerides) accumulates in the liver (steatosis), accompanied by inflammation and fibrosis in non‐alcoholic patients. Lipid accumulation occurs in intracellular vesicles in hepatocytes, typically producing a macrovesicular histologic pattern, and can lead to ballooning degeneration. The injury to hepatocytes can induce inflammation with subsequent fibrosis, and may ultimately lead to cirrhosis and/or hepatocellular carcinoma. A number of rodent models of NASH have been developed and quantification of hepatic lipids in liver sections is a common study endpoint due to the significant role that lipid accumulation plays in NASH pathogenesis. Several histotechnical and morphological methods for measuring hepatic lipid content have been developed. In this study, we used digital image analysis to compare three histological methods for measuring relative lipid burden in liver sections: 1) Oil Red O (ORO) histochemical stain, 2) liver vacuole area by morphometry (digital pathology), and 3) adipophilin expression by immunohistochemistry (IHC). Thirty C57 BL6/J mice were fed a high fat diet for 16 weeks to induce steatosis, followed by 7 weeks of high fat diet feeding in conjunction with daily oral dosing with vehicle or low or high doses of a proprietary compound. After the 23 week study period, livers were collected at necropsy and fixed in formalin before routine processing to paraffin (for IHC or morphology), or cryopreservation and snap‐freezing in OCT compound for cryosectioning (for ORO staining). Serial sections from the left lateral lobe of the liver were prepared from paraffin blocks for hematoxylin and eosin staining (vacuole morphometry) or IHC (for adipophilin), while the ORO stain was performed on cryosections from OCT blocks. Image analysis was performed to measure the area percent of the liver section that was occupied by ORO stain, DAB chromogen (adipophilin IHC) or lipid vacuoles, using Visiopharm image analysis software, and a non‐parametric statistical analysis was performed on the results. There were no significant differences among vehicle, low dose or high dose groups measured using any of the three endpoints for lipid quantification (Kruskal‐Wallis ANOVA); however, the relative patterns of median area percent for the three endpoints were identical among assays (median area percent for low dose compound was highest, and median area percent for high dose compound was lowest), suggesting that the three assays performed similarly, and that any of these assays is adequate for histotechnical measurement of hepatic lipids.Support or Funding InformationPfizerThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Expression of Miscellaneous Genes Upregulated in Hepatocellular Carcinoma in Patients with Alcoholic (ASH) and Non‐alcoholic Steatohepatitis (NASH)
Authors: Lu Jiajie , Nguyen Luan , Samadzadeh Sara , Masouminia Maryam , Mendoza Alejandro , Tillman Brittany , Morgan Timothy , French Barbara , French Samuel 
Published: June 2021

Both non‐alcoholic steatohepatitis (NASH) and alcoholic steatohepatitis (ASH) can lead to cirrhosis and hepatocellular carcinoma. However, the rate of progression to cirrhosis and tumorigenesis in ASH is greater than that in NASH. We asked whether there are differences between the two conditions in the expression levels of proteins involved in the pathogenesis of hepatocellular carcinoma. The proteins tested were presented at the 2017 AASLD Liver Meeting as overexpressed in hepatocellular carcinoma: KLF4, SCL19A1, FANCG, HRH‐1, DNMT1, DNMT3B, TNFR2, DUSP4, EGFR, Integrin, HDACII, and PDE3A. The expression of these proteins was measured in liver biopsy sections from NASH, ASH and normal patients using immunohistochemical staining with fluorescent antibodies and then quantifying the fluorescence intensity morphometrically. In ASH patients, levels of all tested proteins except HRH‐1 were elevated compared to normal patients. In NASH patients, KLF4, SCL19A1, FANCG, DNMT1, DNMT3B, and HDACII levels were increased compared to normal controls while HRH‐1 and PDE3A levels were decreased. The relative expression of all proteins studied except DNMT‐1 was higher in ASH compared to NASH. In conclusion, proteins involved in hepatocellular cancer development are more highly expressed in ASH compared to NASH and normal liver, which corresponds with the higher rate of tumorigenesis in ASH patients compared to NASH patients.Support or Funding InformationThis study was funded by NIH/AAA grant # UO‐21898‐05This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Carbon dioxide challenge enhances the sensitivity of whole body plethysmography (WBP) measurements of lung growth and ozone‐induced changes in juvenile Long‐Evans rats
Authors: Dye Janice , Stewart Erica , Ledbetter Allen , Miller Colette 
Published: June 2021

BackgroundRisk factors associated with development of childhood disorders such as asthma and obesity include prematurity and intrauterine growth restriction. We recently demonstrated that exposure of Long Evans rats in early pregnancy to the air pollutant, ozone, resulted in reduced near‐term fetal growth. We propose to utilize this model of early life environmental stress to examine developmental origins of asthma later in life. Therefore, in this pilot study, we sought to improve the sensitivity of WBP to monitor lung volume growth in rats during development.MethodsSubset A male and female offspring from air‐ or 0.8 ppm ozone‐exposed dams (4h/day on gestational day 5 and 6) (n ≅ 6/group) were monitored weekly from postnatal day (PND) 13 to PND70 for changes in body weight and spontaneous ventilatory parameters [i.e., breathing frequency (f), tidal volume (TV), minute volume (MV), EF50 and Penh]. Data were first obtained using mouse and later rat WBP chambers with EMKA iox 2 software (SCIREQ, Montreal, Canada). To enhance tidal breathing, Subset B male offspring (n = 7; ≥ PND50) were used to optimize a CO2 challenge protocol (5–7.5% CO2) for up to 10 minutes under control conditions, and then later (PND81) to assess whether the CO2 challenge would also be tolerated acutely following exposure to ozone (0.4 or 0.8 ppm ×4h).ResultsSubset A offspring underwent rapid growth during the 8‐week study period with nearly 15‐fold increases in body weight in males and 10‐fold increases in females. Corresponding increases in TV (~0.2 to > 2.0 mL) were observed. In Subset B males, a 6% CO2 challenge for 6 minutes appeared optimal. Likewise, in Subset A rats at PND67, this CO2 challenge resulted in more uniform breathing, less movement‐related artifacts, and increased ventilatory effort (TV increased ~50%); although females were somewhat less responsive. Lastly, use of the 6% CO2 challenge immediately following ozone exposure of Subset B males more readily distinguished the resultant breathing pattern alterations and lung volume deficits occurring.ConclusionsAlthough the CO2‐enhanced lung volume measurements obtained herein are not equivalent to maximal, expiratory maneuvers (i.e., FEV1) as can be achieved in humans, this protocol was well tolerated and may provide a more sensitive, yet non‐invasive method of documenting differential lung growth trajectories in offspring— with or without prior in utero ozone exposure — as they undergo additional exposures during weanling and peri‐adolescent life stages. (Abstract does not reflect USEPA policy).This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Development of a Patient‐Centered Clinical Toxicology Elective in Collaboration with the Illinois Poison Center
Authors: El‐Alfy Abir 
Published: June 2021

Most didactic pharmacy school curricula hardly devote any part of the coursework to toxicology. However, appendix 1 of the 2016 Accreditation Standards lists toxicology topics to be covered in the didactic coursework. These topics include mechanisms, prevention, and treatment of toxicity of drugs and poisons, especially bioterrorism ones. A few hours in didactic curriculum usually satisfy this accreditation requirement. However, since pharmacists play an important role as first health responders, it was essential to develop a clinical toxicology elective course in our program. A 2‐hour course was established in collaboration with clinical pharmacists at the Illinois Poison Center, and offered for third year student pharmacists, during the Spring semester. The course was offered for four years, with average enrollment of 24 students/year. The course topics were tailored yearly according to the American Association of Poison Control Centers' Annual Report (AAPCC). Different active learning techniques were used over the four years. These included case presentations, small group case discussions, in class, and flipped classroom activities. The average student performance in the course for academic years 2011–2014 was 86.77 ± 0.89%. All cases discussed in the course were either provided by the Illinois Poison Center or retrieved from literature case reports. Students analyzed the cases, provided SOAP note reports, and presented them orally. Case presentation grade constituted 25% of the overall grade. The average student performance in the case analysis and presentation was 91.90 ± 0.90%. In conclusion, the toxicology elective developed in collaboration with the poison control center provided a clinically relevant, interactive experience to our students.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Histochemical & Behavioral Analysis of the 3xTg‐AD Mouse Model of Alzheimer's Disease
Authors: Mitrano Darlene A. , Schendzielos Rachel , Croushore Emma , Genovese Benjamin , Quintanilla Ricardo , Dymond Ethan , Pearce Patrick , Houle Samuel , Halleck‐Pinkleton Hanna , Grau Harold , Webb Lisa Smith 
Published: June 2021

Alzheimer's disease (AD) is an incurable neurodegenerative disease that affects about 11% of Americans age 65 and over and has very few FDA‐approved treatments. As the primary areas affected within the brain are the hippocampus and the cortex, the main reported symptoms of this disease are memory loss and confusion. However, the disease can also affect olfactory function, a symptom that develops prior to changes in cognitive function. Due to the lack of effective treatments, developing an animal model that most closely resembles human AD is crucial. The goal of this study was to further validate a mouse model of AD, the 3xTg‐AD mouse which expresses both the amyloid plaques and neurofibrillary tangles in the brain, to determine if the mice express olfactory deficits seen in humans. To do this, we used a buried food test in which mice were food deprived for 24 hours and then placed in a rat cage with a piece of rodent chow buried beneath the bedding. We timed, up to 15 minutes, how long it took the mice to uncover the buried food as a way to measure olfactory detection. The 3xTg‐AD mice were compared to their background and parental strains (C57Bl/6, 129Sv/E, and a hybrid of the C57Bl/6 and 129Sv/E). Preliminary results indicate that over the span of 52 weeks, the 3xTg‐AD mice take significantly longer than the other strains to find the buried food. In order to correlate the behavioral observations with anatomical changes, the Congo Red staining technique was employed to identify whether or not amyloid plaques develop in the olfactory bulbs of the mice. Thus far we have observed plaque accumulation in the glomerular layer of the olfactory bulbs of the 3xTg‐AD mice. Overall, the results of this study increase the validity of the 3xTg‐AD mouse as a model of human AD and leads to new areas of study to determine where else in the brain olfactory function is affected by AD.Support or Funding InformationThis work was supported in part by Award No. 17‐2 from the Commonwealth of Virginia's Alzheimer's and Related Diseases Research Award Fund, administered by the Virginia Center on Aging, School of Allied Health Professions, Virginia Commonwealth University (to D.A. Mitrano & L.S. Webb)This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Angiotensinergic neurotransmission in the medial amygdaloid nucleus modulates the cardiovascular responses to emotional stress in rats.
Authors: Crestani CARLOS C. , Costa‐Ferreira Willian , Gomes‐de‐Souza Lucas 
Published: June 2021

The physiological and behavioral responses observed during aversive threats are coodinated by overlapping circuits in the central nervous system. The medial amygdaloid nucleus (MeA) is a limbic structures implicated in the control of cardiovascular responses to stress. However, the local neurochemical mechanisms involved in such control has not been fully elucidated. In this sense, angiotensin II and their receptors were identified within the MeA. Nevertheless, a possible involvement of this neurochemical mechanism in MeA control of stress‐evoked cardiovascular responses has never been evaluated. Thus, the aim of this study was to investigate the involvement of AT1 receptor within the MeA in control of cardiovascular responses evoked by an acute session of restraint stress in rats. For this, male Wistar rats (240g–260g) had cannula‐guide bilaterally implanted within the MeA. A catheter was also implanted in the femoral artery for mean arterial pressure (MAP) and heart rate (HR) recording. The restraint stress was performed by placing the animals in a plastic cylindrical tube for 60 minutes. Independent group of animals received bilateral microinjections into the MeA of the selective AT1 receptor antagonist losartan (1nmol/100nL), angiotensin II (0.05nmol/100nL) or vehicle (saline, 100nL) 10 min before the onset of the restraint stress session. We observed that bilateral microinjection of losartan into the MeA enhanced the tachycardia evoked by restraint stress (F(1,11)=7, P<0.02), but without affecting the MAP increase (F(1,11)=1, P>0.05). Similarly, bilateral treatment of the Mea with angiotensin II enhanced restraint‐evoked HR increase (F(1,12)=6.5, P<0.02) without affecting the MAP response (F(1,12)=0.5, P>0.05). Taken together, these results provide evidence of an inhibitory role of the AT1 receptor within the MeA in cardiac responses to emotional stress. Besides, results indicate that angiotensin II acting via mechanisms other than the AT1 receptor plays a facilitatory influence on HR response.Support or Funding InformationFAPESP, CNPq and PADC‐FCF/UNESP.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.