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Hwang, Seung Rim, Sook Hee Ku, Min Kyung Joo, Sun Hwa Kim, and Ick Chan Kwon. "Theranostic nanomaterials for image-guided gene therapy." MRS Bulletin 39, no. 1 (January 2014): 44–50. http://dx.doi.org/10.1557/mrs.2013.312.
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Bawazer, Lukmaan A. "Bio Focus: Building synthetic organelles from the gene up." MRS Bulletin 40, no. 1 (January 2015): 9. http://dx.doi.org/10.1557/mrs.2014.315.
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Nakai, T., R. Ishima, H. Sakahara, K. Endo, J. Konishi, and K. Akasaka. "An in vitro1H-MRS Model of Oncogene Transfection." Acta Radiologica 38, no. 6 (November 1997): 1083–86. http://dx.doi.org/10.1080/02841859709172136.
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Purpose: Malignancy is an abnormality of cell division and differentiation based on abnormal expression of oncogenes. This note describes the in vitro1H-NMR spectral features of oncogene-transfected NIH3T3 fibroblast cells compared to non-transfected cells Material and Methods: 1H-NMR spectra of cultured NIH3T3 cells and c-erbB-2 or c-Ha-ras gene-transfected cells were obtained by 400 MHz high resolution NMR. the peaks were assigned by 2D HOHAHA spectra of the cell suspension and the spectral changes were evaluated in 1D and 1D differential spectra Results: the 1H spectra obtained from both transfected cell lines were broadened over all peaks, suggesting reduced mobility in plasma membrane lipid molecules. No other differential spectra for characterizing metabolic change was detected Conclusion: Broadened 1H spectra observed after c-erbB-2 or c-Ha-ras transfection suggest changes of plasma membrane viscosity, which may be related to the oncogene expression
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Mifsud, Karen R., and Johannes M. H. M. Reul. "Acute stress enhances heterodimerization and binding of corticosteroid receptors at glucocorticoid target genes in the hippocampus." Proceedings of the National Academy of Sciences 113, no. 40 (September 21, 2016): 11336–41. http://dx.doi.org/10.1073/pnas.1605246113.
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A stressful event results in secretion of glucocorticoid hormones, which bind to mineralocorticoid receptors (MRs) and glucocorticoid receptors (GRs) in the hippocampus to regulate cognitive and affective responses to the challenge. MRs are already highly occupied by low glucocorticoid levels under baseline conditions, whereas GRs only become substantially occupied by stress- or circadian-driven glucocorticoid levels. Currently, however, the binding of MRs and GRs to glucocorticoid-responsive elements (GREs) within hippocampal glucocorticoid target genes under such physiological conditions in vivo is unknown. We found that forced swim (FS) stress evoked increased hippocampal RNA expression levels of the glucocorticoid-responsive genes FK506-binding protein 5 (Fkbp5), Period 1 (Per1), and serum- and glucocorticoid-inducible kinase 1 (Sgk1). Chromatin immunoprecipitation (ChIP) analysis showed that this stressor caused substantial gene-dependent increases in GR binding and surprisingly, also MR binding to GREs within these genes. Different acute challenges, including novelty, restraint, and FS stress, produced distinct glucocorticoid responses but resulted in largely similar MR and GR binding to GREs. Sequential and tandem ChIP analyses showed that, after FS stress, MRs and GRs bind concomitantly to the same GRE sites within Fkbp5 and Per1 but not Sgk1. Thus, after stress, MRs and GRs seem to bind to GREs as homo- and/or heterodimers in a gene-dependent manner. MR binding to GREs at baseline seems to be restricted, whereas after stress, GR binding may facilitate cobinding of MR. This study reveals that the interaction of MRs and GRs with GREs within the genome constitutes an additional level of complexity in hippocampal glucocorticoid action beyond expectancies based on ligand–receptor interactions.
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Shen, Yao, Gabriel Chan, Michael Xie, Wangyong Zeng, and Liang Liu. "Identification of master regulator genes of UV response and their implications for skin carcinogenesis." Carcinogenesis 40, no. 5 (November 19, 2018): 687–94. http://dx.doi.org/10.1093/carcin/bgy168.
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AbstractSolar UV radiation is a major environmental risk factor for skin cancer. Despite decades of robust and meritorious investigation, our understanding of the mechanisms underlying UV-induced skin carcinogenesis remain incomplete. We previously performed comprehensive transcriptomic profiling in human keratinocytes following exposure to different UV radiation conditions to generate UV-specific gene expression signatures. In this study, we utilized Virtual Inference of Protein Activity by Enriched Regulon (VIPER), a robust systems biology tool, on UV-specific skin cell gene signatures to identify master regulators (MRs) of UV-induced transcriptomic changes. We identified multiple prominent candidate UV MRs, including forkhead box M1 (FOXM1), thyroid hormone receptor interactor 13 and DNA isomerase II alpha, which play important roles in cell cycle regulation and genome stability. MR protein activity was either activated or suppressed by UV in normal keratinocytes. Intriguingly, many of the UV-suppressed MRs were activated in human skin squamous cell carcinomas (SCCs), highlighting their importance in skin cancer development. We further demonstrated that selective inhibition of FOXM1, whose activity was elevated in SCC cells, was detrimental to SCC cell survival. Taken together, our study uncovered novel UV MRs that can be explored as new therapeutic targets for future skin cancer treatment.
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Freitas Ribeiro, Laryssa, Rafael Akira Sato, Andressa de Souza Pollo, Gabriel Augusto Marques Rossi, and Luiz Augusto do Amaral. "Occurrence of Methicillin-Resistant Staphylococcus spp. on Brazilian Dairy Farms that Produce Unpasteurized Cheese." Toxins 12, no. 12 (December 8, 2020): 779. http://dx.doi.org/10.3390/toxins12120779.
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Methicillin-resistant Staphylococcus spp. (MRS) have been identified in several foods, including dairy products. Studies are needed about their occurrence and genetic diversity in the dairy production chain in order to gain a better understanding of their epidemiology and control. This study therefore focuses on isolating and characterizing MRS strains detected in milk used in the production of Brazilian artisanal unpasteurized cheeses. To this end, samples were collected from bovine feces, the hands of milkmen, milking buckets, sieves, unpasteurized milk, whey, water, artisanal unpasteurized cheeses, cheese processing surfaces, cheese handlers, cheese trays, cheese molds, and skimmers at five dairy farms located in the state of São Paulo, Brazil. Colonies suggestive of Staphylococcus spp. were subjected to multiplex PCR to confirm the presence of Staphylococcus aureus and to detect the mecA gene. Sixteen isolates containing mecA gene were detected in samples from unpasteurized cheese and from cheese handlers. None of these isolates were positive to enterotoxin genes. These 16 isolates were subjected to antimicrobial susceptibility tests, which revealed they were resistant to oxacillin, penicillin, and cefepime. Using gene sequencing, the MRS isolates were identified as S. haemolyticus, S. hominis, and S. epidermidis. Furthermore, isolates from cheese handlers’ hands and artisanal unpasteurized cheese presented high genetic similarity by random amplified polymorphic DNA (RAPD-PCR) analysis, which indicates cross contamination during cheese production. Thus, we found that people directly involved in milking and cheese processing activities at small dairy farms are a potential source of contamination of MRS strains in unpasteurized milk and cheese, representing a risk to public health.
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Shovlin, Claire L. "Glaxo/MRS Young Investigator Medal: Molecular Studies on Adenosine Deaminase Deficiency and Hereditary Haemorrhagic Telangiectasia." Clinical Science 94, no. 3 (March 1, 1998): 207–18. http://dx.doi.org/10.1042/cs0940207.
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1. This manuscript describes two different strategies to progress from the clinical assessment of patients to the identification of disease-causing mutations. In the first disease, recognition of a metabolic abnormality allowed direct molecular analysis of the causal gene. In contrast, localization of the second disease gene by linkage analysis was critical to implicate a gene with a previously unsuspected disease role. 2. Two sisters with chronic respiratory disease and recurrent infections were identified as the first cases of adult onset immunodeficiency due to adenosine deaminase deficiency. Autosomal recessive inheritance of two mutations in the adenosine deaminase gene was demonstrated. Enzyme replacement therapy improved the patients' immunological and clinical status. 3. Individuals with pulmonary arteriovenous malformations were used to identify families with hereditary haemorrhagic telangiectasia (HHT, Rendu-Osler-Weber Syndrome). Linkage studies mapped the HHT disease gene in some families to chromosome 9, and demonstrated genetic heterogeneity. The chromosome 9 disease interval was refined, and several candidate genes were assessed. Following the first description of disease-segregating mutations, a complete analysis of the endoglin gene (which encodes an endothelial cell transforming growth factor-β receptor) identified seven novel mutations. Two mutations did not produce mutant mRNA, and disease severity was comparable between families, indicating that HHT results from stoichiometric insufficiency of endoglin. 4. Each study has implications extending beyond the relatively rare disease analysed. The adenosine-deaminase-deficient patients highlight a treatable cause of HIV-negative CD4+ lymphopenia in adults, perhaps accounting for further cases of ‘non-HIV AIDS’. The HHT studies have illuminated a novel area of vascular pathophysiology, with potential relevance to further disease states.
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Trott, Jamie, Yunus Alpagu, Ee Kim Tan, Mohammad Shboul, Yousif Dawood, Michael Elsy, Heike Wollmann, et al. "Mitchell-Riley syndrome iPSCs exhibit reduced pancreatic endoderm differentiation due to a mutation in RFX6." Development 147, no. 21 (October 8, 2020): dev194878. http://dx.doi.org/10.1242/dev.194878.
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ABSTRACTMitchell-Riley syndrome (MRS) is caused by recessive mutations in the regulatory factor X6 gene (RFX6) and is characterised by pancreatic hypoplasia and neonatal diabetes. To determine why individuals with MRS specifically lack pancreatic endocrine cells, we micro-CT imaged a 12-week-old foetus homozygous for the nonsense mutation RFX6 c.1129C>T, which revealed loss of the pancreas body and tail. From this foetus, we derived iPSCs and show that differentiation of these cells in vitro proceeds normally until generation of pancreatic endoderm, which is significantly reduced. We additionally generated an RFX6HA reporter allele by gene targeting in wild-type H9 cells to precisely define RFX6 expression and in parallel performed in situ hybridisation for RFX6 in the dorsal pancreatic bud of a Carnegie stage 14 human embryo. Both in vitro and in vivo, we find that RFX6 specifically labels a subset of PDX1-expressing pancreatic endoderm. In summary, RFX6 is essential for efficient differentiation of pancreatic endoderm, and its absence in individuals with MRS specifically impairs formation of endocrine cells of the pancreas head and tail.
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Lee, Eun-Jae, Mi-Sun Oh, Jong S. Kim, Dae-Il Chang, Jong-Ho Park, Jae-Kwan Cha, Ji Hoe Heo, et al. "Serotonin transporter gene polymorphisms may be associated with poststroke neurological recovery after escitalopram use." Journal of Neurology, Neurosurgery & Psychiatry 89, no. 3 (October 13, 2017): 271–76. http://dx.doi.org/10.1136/jnnp-2017-316882.
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ObjectiveSelective serotonin reuptake inhibitors (SSRIs) putatively improve neurological recovery after stroke. We aimed to investigate whether serotonin transporter (SERT) gene polymorphisms are related to the responsiveness to SSRIs in the poststroke neurological recovery.MethodsThis was a post hoc analysis of the EMOTION study (ClinicalTrials.gov NCT01278498), a randomised, placebo-controlled, double-blind trial examining the efficacy of escitalopram on emotional and neurological disturbances after acute stroke. Patients with no/minimal disability initially (modified Rankin Scale (mRS) 0–1) were excluded. Of the participants, 301 underwent genetic studies of the STin2 (a variable number tandem repeat (VNTR) in intron 2) (STin2 12/10 and STin2 12/12 genotypes) and 5-HTTLPR (a variable-length repeat in the promoter region) polymorphisms of SERT. We explored whether neurological function (National Institutes of Health Stroke Scale (NIHSS) score and mRS) at 3 months would differ according to SERT polymorphisms within each treatment arm (escitalopram and placebo).ResultsAmong the escitalopram users (n=159), neurological function in subjects with STin2 12/10 (n=29) improved significantly more than that in STin2 12/12 carriers (n=130) at 3 months. After adjusting for age, initial NIHSS and depression, STin2 12/10 independently predicted a good clinical outcome (mRS 0–1) (OR 2.99, 95% CI 1.04 to 8.58) at 3 months. However, differences between STin2 polymorphisms were not shown in the placebo group (n=142). 5-HTTLPR polymorphisms were not associated with neurological recovery in any treatment group.ConclusionSTin2 VNTR polymorphisms may be associated with poststroke neurological recovery after SSRI therapy. Further studies are needed to identify the role of serotonin in neurological recovery after stroke.
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Ngassam Tchamba, Cyrille, Jean-Noël Duprez, Pierrick Lucas, Yannick Blanchard, Filip Boyen, Freddy Haesebrouck, Maria A. Argudín, Jacques Mainil, and Damien Thiry. "Comparison of the Staphylococcal Chromosome Cassette (SCC) mec in Methicillin-Resistant Staphylococcus aureus (MRSA) and Non-aureus Staphylococci (MRNAS) from Animals and Humans." Antibiotics 10, no. 3 (March 4, 2021): 256. http://dx.doi.org/10.3390/antibiotics10030256.
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Methicillin-resistant Staphylococcus aureus (MRSA) and non-aureus staphylococci (MRNAS) cause different infections in animals, including mastitis, in livestock and humans. This study aimed to identify and compare the staphylococcal chromosome cassette mec (SCCmec) types of MRSA or MRNAS isolated from several animal species and humans in different countries. Of 1462 S. aureus and non-aureus staphylococci, 68 grew on Chrom MRSA ID® agar, were phenotypically resistant to cefoxitin and tested positive with the PCR for the mecA gene. These 60 MRSA and 8 MRNAS were isolated in Belgium mainly from cows (livestock-associated (LA) MRS) and humans (community-acquired (CA) MRS) and in Japan from dogs and cats. The SCCmec cassettes were identified by multiplex PCR in 52 MRSA and 7 MRNAS and by whole genome sequencing (WGS) in 8 additional MRSA. The SCCmec types IV and V were the most frequent in Belgian LA-MRS and CA-MRS, while the SCCmec type II was identified in four of the five Japanese MRSA. The remaining isolate was a bovine S. haemolyticus in which no SCCmec was identified. These results confirm the high prevalence of the SCCmec types IV and V in LA-MRS and CA-MRS in Belgium, emphasizing the possible public health hazard of the former, and the absence of SCCmec in some MRNAS.
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Oon, Ming Liang, Jing Quan Lim, Bernett Lee, Sai Mun Leong, Gwyneth Shook-Ting Soon, Zi Wei Wong, Evelyn Huizi Lim, et al. "T-Cell Lymphoma Clonality by Copy Number Variation Analysis of T-Cell Receptor Genes." Cancers 13, no. 2 (January 19, 2021): 340. http://dx.doi.org/10.3390/cancers13020340.
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T-cell lymphomas arise from a single neoplastic clone and exhibit identical patterns of deletions in T-cell receptor (TCR) genes. Whole genome sequencing (WGS) data represent a treasure trove of information for the development of novel clinical applications. However, the use of WGS to identify clonal T-cell proliferations has not been systematically studied. In this study, based on WGS data, we identified monoclonal rearrangements (MRs) of T-cell receptors (TCR) genes using a novel segmentation algorithm and copy number computation. We evaluated the feasibility of this technique as a marker of T-cell clonality using T-cell lymphomas (TCL, n = 44) and extranodal NK/T-cell lymphomas (ENKTLs, n = 20), and identified 98% of TCLs with one or more TCR gene MRs, against 91% detected using PCR. TCR MRs were absent in all ENKTLs and NK cell lines. Sensitivity-wise, this platform is sufficiently competent, with MRs detected in the majority of samples with tumor content under 25% and it can also distinguish monoallelic from biallelic MRs. Understanding the copy number landscape of TCR using WGS data may engender new diagnostic applications in hematolymphoid pathology, which can be readily adapted to the analysis of B-cell receptor loci for B-cell clonality determination.
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Subramani, Elavarasan, Chloe Najac, Georgios Batsios, Pavithra Viswanath, Marina Radoul, Anne Marie Gillespie, Russell O. Pieper, and Sabrina Ronen. "EXTH-20. HYPERPOLARIZED [2-13C] PYRUVATE TO [5-13C] GLUTAMATE AS BIOMARKERS OF IDH1 MUTANT GLIOMA RESPONSE TO TEMOZOLOMIDE THERAPY." Neuro-Oncology 21, Supplement_6 (November 2019): vi86. http://dx.doi.org/10.1093/neuonc/noz175.354.
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Abstract Low-grade gliomas, driven by mutations in the cytosolic isocitrate dehydrogenase 1 (IDH1) gene, are less aggressive than primary glioblastoma, but nonetheless always recur and ultimately lead to patient death. The treatment of IDH1 mutant patients with Temozolomide (TMZ) improves survival, but there remains a need for complementary imaging methods to assess response to therapy at an early time point. The goal of this study was, therefore, to determine the value of magnetic resonance spectroscopy (MRS)-based metabolic imaging biomarkers for detection of response to treatment. To this end we investigated NHA and U87 cells expressing IDH1 R132H mutant gene (NHAIDHmut and U87IDHmut) and first used 1H MRS combined with chemometrics to examined the metabolic alterations that occurred following treatment with the IC50 value of TMZ. We observed a significant increase in 2-hydroxyglutarate (2-HG), glutamate, and glutamine, and metabolic pathway analysis showed tricarboxylic acid (TCA) cycle and pyruvate metabolism to be significantly altered pathways following TMZ treatment compared to DMSO control. To confirm changes in TCA cycle flux and to assess the metabolic pathways contributing to the increase in 2-HG and glutamate/glutamine, cells were then labelled with [1-13C] glucose and [3-13C] glutamine. Our data indicated that both glucose flux via the TCA to glutamate and 2HG, and the contribution of glutamine to glutamate and 2HG were increased following TMZ treatment. Finally, we used hyperpolarized 13C-MRS to dynamically probe the metabolism of hyperpolarized [2-13C] pyruvate and its conversion to hyperpolarized [5-13C] glutamate via the TCA cycle. Consistent with our previous findings, we observed that hyperpolarized [5-13C] glutamate synthesis was significantly higher in TMZ-treated cells compared to controls. Collectively, our findings identify 1H MRS-detectable elevation of 2-HG and glutamate/glutamine as well as hyperpolarized 13C-MRS-detectable [5-13C] glutamate production from [2-13C] pyruvate as potentially translatable metabolic biomarkers of response to TMZ therapy in mutant IDH1 glioma.
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Choi, Changho, Jack M. Raisanen, Sandeep K. Ganji, Song Zhang, Sarah S. McNeil, Zhongxu An, Akshay Madan, et al. "Prospective Longitudinal Analysis of 2-Hydroxyglutarate Magnetic Resonance Spectroscopy Identifies Broad Clinical Utility for the Management of Patients With IDH-Mutant Glioma." Journal of Clinical Oncology 34, no. 33 (November 20, 2016): 4030–39. http://dx.doi.org/10.1200/jco.2016.67.1222.
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Purpose Proton magnetic resonance spectroscopy (MRS) of the brain can detect 2-hydroxyglutarate (2HG), the oncometabolite produced in neoplasms harboring a mutation in the gene coding for isocitrate dehydrogenase ( IDH). We conducted a prospective longitudinal imaging study to determine whether quantitative assessment of 2HG by MRS could serve as a noninvasive clinical imaging biomarker for IDH-mutated gliomas. Patients and Methods 2HG MRS was performed in 136 patients using point-resolved spectroscopy at 3 T in parallel with standard clinical magnetic resonance imaging and assessment. Data were analyzed in patient cohorts representing the major phases of the glioma clinical course and were further subgrouped by histology and treatment type to evaluate 2HG. Histologic correlations were performed. Results Quantitative 2HG MRS was technically and biologically reproducible. 2HG concentration > 1 mM could be reliably detected with high confidence. During the period of indolent disease, 2HG concentration varied by less than ± 1 mM, and it increased sharply with tumor progression. 2HG concentration was positively correlated with tumor cellularity and significantly differed between high- and lower-grade gliomas. In response to cytotoxic therapy, 2HG concentration decreased rapidly in 1p/19q codeleted oligodendrogliomas and with a slower time course in astrocytomas and mixed gliomas. The magnitude and time course of the decrease in 2HG concentration and magnitude of the decrease in tumor volume did not differ between oligodendrogliomas treated with temozolomide or carmustine. Criteria for 2HG MRS were established to make a presumptive molecular diagnosis of an IDH mutation in gliomas technically unable to undergo a surgical procedure. Conclusion 2HG concentration as measured by MRS was reproducible and reliably reflected the disease state. These data provide a basis for incorporating 2HG MRS into clinical management of IDH-mutated gliomas.
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Zielman, R., WM Teeuwisse, F. Bakels, J. Van der Grond, A. Webb, MA van Buchem, MD Ferrari, MC Kruit, and GM Terwindt. "Biochemical changes in the brain of hemiplegic migraine patients measured with 7 tesla 1H-MRS." Cephalalgia 34, no. 12 (March 20, 2014): 959–67. http://dx.doi.org/10.1177/0333102414527016.
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Aim The aim of this study was to assess biochemical changes in the brain of patients with hemiplegic migraine in between attacks. Methods Eighteen patients with hemiplegic migraine (M:F, 7:11; age 38 ± 14 years) of whom eight had a known familial hemiplegic migraine (FHM) mutation (five in the CACNA1A gene (FHM1), three in the ATP1A2 gene (FHM2)) and 19 age- and sex-matched healthy controls (M:F, 7:12; mean age 38 ± 12 years) were studied. We used single-voxel 7 tesla 1H-MRS (STEAM, TR/TM/TE = 2000/19/21 ms) to investigate four brain regions in between attacks: cerebellum, hypothalamus, occipital lobe, and pons. Results Patients with hemiplegic migraine showed a significantly lower total N-acetylaspartate/total creatine ratio (tNAA/tCre) in the cerebellum (median 0.73, range 0.59–1.03) than healthy controls (median 0.79, range (0.67–0.95); p = 0.02). In FHM1 patients with a CACNA1A mutation, the tNAA/tCre was lowest. Discussion We found a decreased cerebellar tNAA/tCre ratio that might serve as an early biomarker for neuronal dysfunction and/or loss. This is the first high-spectral resolution 7 tesla 1H-MRS study of interictal biochemical brain changes in hemiplegic migraine patients.
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Bassuk, A. G., A. Joshi, B. K. Burton, M. B. Larsen, D. M. Burrowes, and C. Stack. "Alexander disease with serial MRS and a new mutation in the glial fibrillary acidic protein gene." Neurology 61, no. 7 (October 13, 2003): 1014. http://dx.doi.org/10.1212/01.wnl.0000082440.42354.d0.
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Li, Shuo, Jiafang Li, Nan Wang, Gaixiang Hao, and Jinsheng Sun. "Characterization of UDP-Activated Purinergic Receptor P2Y6 Involved in Japanese Flounder Paralichthys olivaceus Innate Immunity." International Journal of Molecular Sciences 19, no. 7 (July 19, 2018): 2095. http://dx.doi.org/10.3390/ijms19072095.
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Uridine 5’-diphosphate (UDP)-activated purinergic receptor P2Y6 is a member of a G-protein-coupled purinergic receptor family that plays an important role in mammalian innate immunity. However, the role of the P2Y6 receptor (P2Y6R) in fish immunity has not been investigated. In this report, we characterized a P2Y6R gene from Japanese flounder (Paralichthys olivaceus) and examined its role in fish innate immunity. Sequence analysis reveals that the Japanese flounder P2Y6R protein is conserved and possesses four potential glycosylation sites. Quantitative real-time RT-PCR analysis shows that P2Y6R is broadly distributed in all examined Japanese flounder tissues with dominant expression in the liver. In addition, P2Y6R gene expression was up-regulated in head kidney macrophages (HKMs) upon lipopolysaccharides (LPS) and poly(I:C) stimulations but down-regulated by LPS challenge in peripheral blood leukocytes (PBLs). Furthermore, pharmacological inhibition of the endogenous P2Y6 receptor activity by the potently selective P2Y6R antagonist, MRS 2578, greatly up-regulated pro-inflammatory cytokine interleukin (IL)-1β, IL-6 and TNF-α gene expression in PBL cells treated with UDP. Moreover, LPS- and poly(I:C)-induced gene expression of IL-1β and TNF-α in Japanese flounder PBL cells was attenuated significantly by inhibition of P2Y6R activity with antagonist MRS 2578. Collectively, we, for the first time, showed the involvement of functional purinergic P2Y6R in fish innate immunity.
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Smeianov, Vladimir V., Patrick Wechter, Jeffery R. Broadbent, Joanne E. Hughes, Beatriz T. Rodríguez, Tove K. Christensen, Ylva Ardö, and James L. Steele. "Comparative High-Density Microarray Analysis of Gene Expression during Growth of Lactobacillus helveticus in Milk versus Rich Culture Medium." Applied and Environmental Microbiology 73, no. 8 (February 23, 2007): 2661–72. http://dx.doi.org/10.1128/aem.00005-07.
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ABSTRACT Lactobacillus helveticus CNRZ32 is used by the dairy industry to modulate cheese flavor. The compilation of a draft genome sequence for this strain allowed us to identify and completely sequence 168 genes potentially important for the growth of this organism in milk or for cheese flavor development. The primary aim of this study was to investigate the expression of these genes during growth in milk and MRS medium by using microarrays. Oligonucleotide probes against each of the completely sequenced genes were compiled on maskless photolithography-based DNA microarrays. Additionally, the entire draft genome sequence was used to produce tiled microarrays in which noninterrupted sequence contigs were covered by consecutive 24-mer probes and associated mismatch probe sets. Total RNA isolated from cells grown in skim milk or in MRS to mid-log phase was used as a template to synthesize cDNA, followed by Cy3 labeling and hybridization. An analysis of data from annotated gene probes identified 42 genes that were upregulated during the growth of CNRZ32 in milk (P < 0.05), and 25 of these genes showed upregulation after applying Bonferroni's adjustment. The tiled microarrays identified numerous additional genes that were upregulated in milk versus MRS. Collectively, array data showed the growth of CNRZ32 in milk-induced genes encoding cell-envelope proteinases, oligopeptide transporters, and endopeptidases as well as enzymes for lactose and cysteine pathways, de novo synthesis, and/or salvage pathways for purines and pyrimidines and other functions. Genes for a hypothetical phosphoserine utilization pathway were also differentially expressed. Preliminary experiments indicate that cheese-derived, phosphoserine-containing peptides increase growth rates of CNRZ32 in a chemically defined medium. These results suggest that phosphoserine is used as an energy source during the growth of L. helveticus CNRZ32.
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McDade, Eric M., Bradley F. Boeve, Julie A. Fields, Neeraj Kumar, Rosa Rademakers, Matt C. Baker, BSc David S. Knopman, Ronald C. Petersen, Clifford R. Jack, and Kejal Kantarci. "MRS in Early and Presymptomatic Carriers of a Novel Octapeptide Repeat Insertion in the Prion Protein Gene." Journal of Neuroimaging 23, no. 3 (May 21, 2012): 409–13. http://dx.doi.org/10.1111/j.1552-6569.2012.00717.x.
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Donnelly, Mark K., Elizabeth A. Crago, Yvette P. Conley, Jeffery R. Balzer, Dianxu Ren, Andrew F. Ducruet, Patrick M. Kochanek, Paula R. Sherwood, and Samuel M. Poloyac. "20-HETE is Associated with Unfavorable Outcomes in Subarachnoid Hemorrhage Patients." Journal of Cerebral Blood Flow & Metabolism 35, no. 9 (April 29, 2015): 1515–22. http://dx.doi.org/10.1038/jcbfm.2015.75.
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Emerging evidence has suggested that patients experiencing aneurysmal subarachnoid hemorrhage (aSAH) develop vascular dysregulation as a potential contributor to poor outcomes. Preclinical studies have implicated the novel microvascular constrictor, 20-hydroxyeicosatetraenoic acid (20-HETE) in aSAH pathogenesis, yet the translational relevance of 20-HETE in patients with aSAH is largely unknown. The goal of this research was to determine the relationship between 20-HETE cerebrospinal fluid (CSF) levels, gene variants in 20-HETE synthesis, and acute/long-term aSAH outcomes. In all, 363 adult patients (age 18 to 75) with aSAH were prospectively recruited from the University of Pittsburgh Medical Center neurovascular Intensive Care Unit. Patients were genotyped for polymorphic variants and cytochrome P450 (CYP)-eicosanoid CSF levels were measured over 14 days. Outcomes included delayed cerebral ischemia (DCI), clinical neurologic deterioration (CND), and modified Rankin Scores (MRS) at 3 and 12 months. Patients with CND and unfavorable 3-month MRS had 2.2- and 2.7-fold higher mean 20-HETE CSF levels, respectively. Patients in high/moderate 20-HETE trajectory groups (35.7%) were 2.5-, 2.1-, 3.1-, 3.3-, and 2.1-fold more likely to have unfavorable MRS at 3 months, unfavorable MRS at 12 months, mortality at 3 months, mortality at 12 months, and CND, respectively. These results showed that 20-HETE is associated with acute and long-term outcomes and suggest that 20-HETE may be a novel target in aSAH.
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Koning, Anne-Sophie C. A. M., Jacobus C. Buurstede, Lisa T. C. M. van Weert, and Onno C. Meijer. "Glucocorticoid and Mineralocorticoid Receptors in the Brain: A Transcriptional Perspective." Journal of the Endocrine Society 3, no. 10 (July 24, 2019): 1917–30. http://dx.doi.org/10.1210/js.2019-00158.
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Abstract Adrenal glucocorticoid hormones are crucial for maintenance of homeostasis and adaptation to stress. They act via the mineralocorticoid receptors (MRs) and glucocorticoid receptors (GRs)—members of the family of nuclear receptors. MRs and GRs can mediate distinct, sometimes opposite, effects of glucocorticoids. Both receptor types can mediate nongenomic steroid effects, but they are best understood as ligand-activated transcription factors. MR and GR protein structure is similar; the receptors can form heterodimers on the DNA at glucocorticoid response elements (GREs), and they share a number of target genes. The transcriptional basis for opposite effects on cellular physiology remains largely unknown, in particular with respect to MR-selective gene transcription. In this review, we discuss proven and potential mechanisms of transcriptional specificity for MRs and GRs. These include unique GR binding to “negative GREs,” direct binding to other transcription factors, and binding to specific DNA sequences in conjunction with other transcription factors, as is the case for MRs and NeuroD proteins in the brain. MR- and GR-specific effects may also depend on specific interactions with transcriptional coregulators, downstream mediators of transcriptional receptor activity. Current data suggest that the relative importance of these mechanisms depends on the tissue and physiological context. Insight into these processes may not only allow a better understanding of homeostatic regulation but also the development of drugs that target specific aspects of disease.
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Morse, David L., Danielle Carroll, Sam Day, Heather Gray, Pooja Sadarangani, Shiva Murthi, Constantin Job, Brenda Baggett, Natarajan Raghunand, and Robert J. Gillies. "Characterization of breast cancers and therapy response by MRS and quantitative gene expression profiling in the choline pathway." NMR in Biomedicine 22, no. 1 (November 18, 2008): 114–27. http://dx.doi.org/10.1002/nbm.1318.
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Stegman, Lauren D., Oded Ben-Yoseph, James P. Freyer, and Brian D. Ross. "In Vivo31P MRS Evaluation of Ganciclovir Toxicity in C6 Gliomas Stably Expressing the Herpes Simplex Thymidine Kinase Gene." NMR in Biomedicine 9, no. 8 (December 1996): 364–68. http://dx.doi.org/10.1002/(sici)1099-1492(199612)9:8<364::aid-nbm436>3.0.co;2-w.
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Wolfe, John H., Satish L. Deshmane, and Nigel W. Fraser. "Herpesvirus vector gene transfer and expression of β–glucuronidase in the central nervous system of MRS VII mice." Nature Genetics 1, no. 5 (August 1992): 379–84. http://dx.doi.org/10.1038/ng0892-379.
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Kotronen, Anna, Hannele Yki-Järvinen, Anna Aminoff, Robert Bergholm, Kirsi H. Pietiläinen, Jukka Westerbacka, Philippa J. Talmud, et al. "Genetic variation in the ADIPOR2 gene is associated with liver fat content and its surrogate markers in three independent cohorts." European Journal of Endocrinology 160, no. 4 (April 2009): 593–602. http://dx.doi.org/10.1530/eje-08-0900.
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AimsWe investigated whether polymorphisms in candidate genes involved in lipid metabolism and type 2 diabetes are related to liver fat content.MethodsLiver fat content was measured using proton magnetic resonance spectroscopy (1H-MRS) in 302 Finns, in whom single nucleotide polymorphisms (SNPs) in acyl-CoA synthetase long-chain family member 4 (ACSL4), adiponectin receptors 1 and 2 (ADIPOR1andADIPOR2), and the three peroxisome proliferator-activated receptors (PPARA,PPARD, andPPARG) were analyzed. To validate our findings, SNPs significantly associated with liver fat content were studied in two independent cohorts and related to surrogate markers of liver fat content.ResultsIn the Finnish subjects, polymorphisms inACSL4(rs7887981),ADIPOR2(rs767870), andPPARG(rs3856806) were significantly associated with liver fat content measured with1H-MRS after adjusting for age, gender, and BMI. Anthropometric and circulating parameters were comparable between genotypes. In the first validation cohort of ∼ 600 Swedish men,ACSL4rs7887981 was related to fasting insulin and triglyceride concentrations, andADIPOR2rs767870 to serum γ glutamyltransferase concentrations after adjusting for BMI. The SNP inPPARG(rs3856806) was not significantly associated with any relevant metabolic parameter in this cohort. In the second validation cohort of ∼3000 subjects from Western Finland,ADIPOR2rs767870, but notACSL4rs7887981 was related to fasting triglyceride concentrations.ConclusionsGenetic variation, particularly in theADIPOR2gene, contributes to variation in hepatic fat accumulation in humans.
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JIN, JUNHUA, LINXIA JIE, HANWEI ZHANG, YUANHONG XIE, HUI LIU, XIUZHI GAO, and HONGXING ZHANG. "Pediocin AcH Is Transcriptionally Regulated by a Two-Component System in Lactobacillus plantarum subsp. plantarum." Journal of Food Protection 83, no. 10 (May 18, 2020): 1693–700. http://dx.doi.org/10.4315/jfp-19-587.
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ABSTRACT The quorum-sensing regulation of class II bacteriocin (AcH) synthesis in Lactobacillus plantarum subsp. plantarum Zhang-LL was studied. No detectable inhibition zone was formed by the supernatant of L. plantarum subsp. plantarum Zhang-LL culture in skim milk (SM) with an inoculum size of 7 × 102 CFU/mL after incubation for 36 h. Hence, this culture system was used to investigate the induced regulation mechanism of bacteriocin production in L. plantarum subsp. plantarum Zhang-LL. Bacteriocin production by this bacterium in SM medium was induced by treatment with inactivated culture supernatant from de Man Rogosa Sharpe (MRS) medium (supernatant-MRS). Pediocin AcH encoded by the papA gene in a plasmid in strain Zhang-LL was the inducer present in supernatant-MRS. This is the first report of the role of pediocin AcH in the quorum-sensing regulation of class II bacteriocin synthesis. The mRNA of the papA, papB, papC, and papD genes involved in bacteriocin synthesis by strain Zhang-LL in SM medium was upregulated significantly after being induced by pediocin AcH. This study offers the first evidence that the ABT40_05745, ABT40_05750, and ABT40_11975 components of two-component systems in L. plantarum subsp. plantarum Zhang-LL are involved in the induced regulation of AcH bacteriocin production. HIGHLIGHTS
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Elnageh, Hiam R., Murad A. Hiblu, Mohamed Salah Abbassi, Yousef M. Abouzeed, and Mohamed O. Ahmed. "Prevalence and antimicrobial resistance of Staphylococcus species isolated from cats and dogs." Open Veterinary Journal 10, no. 4 (February 5, 2021): 452–56. http://dx.doi.org/10.4314/ovj.v10i4.13.
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Background: Methicillin-resistant staphylococci (MRS) are an emerging global problem with serious public health concern.Aims: This study investigated the prevalence and antimicrobial susceptibility of commensal Staphylococcus species isolated from healthy and clinical cats and dogs.Methods: Nasal swab samples were collected from animals and processed using selective and semi-selective mediums. Presumptive isolates were subjected to biochemical testing and analyzed using the Phoenix automated identification and susceptibility testing system. PCRs protocols were used to screen for mecA and pvl genes.Results: In total, 151 pets (103 cats and 48 dogs) were enrolled, of which 14 dogs (29%) and 24 cats (23%) were colonized with various Staphylococcus species mainly originated from healthy animals. A total of 38 staphylococci isolates were collected and distributed between 24 coagulase-negative and 14 coagulase-positive staphylococci. Only 13 staphylococci strains were identified as MRS, out of which only five isolates expressed that the mecA gene exclusively originated from healthy pets.Conclusion: This is the first study reporting the prevalence and colonization status of staphylococci species and MRS strains isolated from cats and dogs in Libya. The study reports important information of medical and clinical importance on antimicrobial and multidrug resistance of different staphylococci strains, particularly the coagulase negative species.
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Dossi, Gabriele, Letizia Squarcina, and Mario Rango. "In Vivo Mitochondrial Function in Idiopathic and Genetic Parkinson’s Disease." Metabolites 10, no. 1 (December 28, 2019): 19. http://dx.doi.org/10.3390/metabo10010019.
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Parkinson’s disease (PD) is associated with brain mitochondrial dysfunction. High-energy phosphates (HEPs), which rely on mitochondrial functioning, may be considered potential biomarkers for PD. Phosphorus magnetic resonance spectroscopy (31P-MRS) is a suitable tool to explore in vivo cerebral energetics. We considered 10 31P-MRS studies in order to highlight the main findings about brain energetic compounds in patients affected by idiopathic PD and genetic PD. The studies investigated several brain areas such as frontal lobes, occipital lobes, temporoparietal cortex, visual cortex, midbrain, and basal ganglia. Resting-state studies reported contrasting results showing decreased as well as normal or increased HEPs levels in PD patients. Functional studies revealed abnormal PCr + βATP levels in PD subjects during the recovery phase and abnormal values at rest, during activation and recovery in one PD subject with PINK1 gene mutation suggesting that mitochondrial machinery is more impaired in PD patients with PINK1 gene mutation. PD is characterized by energetics impairment both in idiopathic PD as well as in genetic PD, suggesting that mitochondrial dysfunction underlies the disease. Studies are still sparse and sometimes contrasting, maybe due to different methodological approaches. Further studies are needed to better assess the role of mitochondria in the PD development.
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Rađenović, Milan, Jelena Ašanin, Ksenija Aksentijević, and Dušan Mišić. "INVESTIGATION OF PRESENCE OF METHICILLIN RESISTANT STAPHYLOCOCCI IN STUDENTS OF THE FACULTY OF VETERINARY MEDICINE AT THE UNIVERSITY OF BELGRADE." Archives of Veterinary Medicine 9, no. 2 (March 21, 2017): 17–28. http://dx.doi.org/10.46784/e-avm.v9i2.86.
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Resistance to methicillin in staphylococci is considered to be one of the most dangerous forms of bacterial resistances to antibiotics. Methicillinresistant staphylococci (MRS) are zoonotic agents which cause local and systemic infections in humans and animals, oft en with a fatal outcome due to the absence of adequate antibiotic therapy. People colonized with strains of MRS are asymptomatic carriers and reservoirs of these strains in human populations. Th e aim of this research was to determine the prevalence of strains of MRS among clinically healthy students of the Faculty of Veterinary Medicine in Belgrade. Th e study was conducted on 100 volunteers: 62 males and 38 females. Given that staphylococci are expected to be found in the highest percentage in the nose and on the armpit skin, the swabs were taken from these regions of each person. Blood agar was innoculated immediately on taking the swabs Aft er the incubation and isolation, the staphylococci were identifi ed to species level. Their susceptibility to methicillin was tested in a disk-diff usion test with cefoxitin. All strains which were found to be resistant to cefoxitin were investigated for the presence of mecA gene with PCR. Staphylococci were isolated in 146 out of the 200 swabs taken: there were 79 nose swabs and 67 axillar swabs positive for these bacteria. Seventeen isolates were resistant to cefoxitin and the presence of the mecA gene was confi rmed in seven, four of which were taken from the nose and three from the axillary region. The results of this research show that, being 6%, the prevalence of mecA-positive staphylococci in the population of clinically healthy students of veterinary medicine is significant. Th e percentage of methicillin-resistant staphylococci was higher in nose than in the axillar region of the students.
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Pavisic, Jovana, Chankrit Sethi, Chris Jones, Stergios Zacharoulis, and Andrea Califano. "DIPG-40. TARGETING MASTER REGULATOR DEPENDENCIES IN DIFFUSE INTRINSIC PONTINE GLIOMA (DIPG)." Neuro-Oncology 22, Supplement_3 (December 1, 2020): iii294—iii295. http://dx.doi.org/10.1093/neuonc/noaa222.087.
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Abstract Diffuse intrinsic pontine glioma (DIPG) remains a fatal disease with no effective drugs to date. Mutation-based precision oncology approaches are limited by lack of targetable mutations and genetic heterogeneity. We leveraged systems biology methodologies to discover common targetable disease drivers—master regulator proteins (MRs)—in DIPG to expand treatment options. Using the metaVIPER algorithm, we interrogated an integrated low grade glioma and GBM gene regulatory network with 31 DIPG-gene expression signatures to identify tumor-specific MRs by differential expression of their transcriptional targets. Unsupervised clustering identified MR signatures of upregulated activity in RRM2/TOP2A in 13 patients, CD3D in 5 patients, and MMP7, TACSTD2, RAC2 and SLC15A1/SLC34A2 in individual patients, all of which can be targeted. Notably, intratumoral administration of etoposide by convection enhanced delivery was effective in murine proneural gliomas in which TOP2 was identified as a MR while RRM2—targetable by drugs such as cladribine—has been shown to be a positive regulator of glioma progression whose knock-down inhibits tumor growth. We also prioritized drugs by their ability to reverse MR-activity signatures using a large drug-perturbation database. Patients clustered by predicted drug sensitivities with distinct groups of tumors predicted to respond to proteasome inhibitors, Thiotepa or Volasertib all of which have early evidence in treating gliomas. We will refine this analysis in a multi-institutional study of >100 patient gene expression profiles to define MR signatures driving known biological/molecular disease subtypes, use DIPG cell lines recapitulating common MR architectures to optimize therapy prioritization, and validate our findings in vivo.
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Elmoslemany, Ahmed, Ibrahim Elsohaby, Mohammed Alorabi, Mohamed Alkafafy, Theeb Al-Marri, Ali Aldoweriej, Fanan A. Alaql, Abdullah Almubarak, and Mahmoud Fayez. "Diversity and Risk Factors Associated with Multidrug and Methicillin-Resistant Staphylococci Isolated from Cats Admitted to a Veterinary Clinic in Eastern Province, Saudi Arabia." Antibiotics 10, no. 4 (March 31, 2021): 367. http://dx.doi.org/10.3390/antibiotics10040367.
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Understanding the distribution, antimicrobial resistance (AMR), and risk factors associated with multidrug-resistant (MDR) and methicillin-resistant staphylococci (MRS) isolated from cats admitted to veterinary clinics may decrease the risk of MDR and MRS transmission to humans and other cats. As such, the objectives of this study were to investigate the diversity in Staphylococcus spp. recovered from different anatomical locations in healthy and diseased cats and to determine the occurrence of MDR and MRS spp. as well as possible risk factors associated with colonization in these cats. Five swabs were collected from the anus, skin, ear canal, conjunctival sac, and nares of each cat (209 healthy and 191 diseased) admitted to a veterinary clinic in Eastern Province, Saudi Arabia, between January and December 2018. Prior to sample collection, cat owners completed a questionnaire collecting information on cat demographics, health status, management, and antimicrobial usage. In total, 179 Staphylococcus isolates were recovered from healthy (n = 71) and diseased (n = 108) cats, including 94 (52.5%) coagulase-positive staphylococci (CoPS), and 85 (47.5%) coagulase-negative staphylococci (CoNS). Five Staphylococcus spp. were identified, namely, Staphylococcus aureus, Staphylococcus pseudintermedius, Staphylococcus felis, Staphylococcus capitis, and Staphylococcus saprophyticus. Staphylococcus isolates were most commonly resistant to penicillin (56.4%) and ciprofloxacin (25.7%); however, no isolate was resistant to clindamycin. Thirty (16.8%) Staphylococcus spp. (24 S. aureus and 6 S. pseudintermedius) isolates were MDR, with resistance to up to six different antibiotic classes. Only 17 (9.5%) Staphylococcus spp. (15 methicillin-resistant S. aureus and 2 methicillin-resistant S. pseudintermedius) harbored the mecA gene. Risk factor analysis showed that cats with a history of antibiotic therapy, those raised mainly indoors with a child, and those who visit a veterinary clinic for treatment were at higher risk of MDR and MRS colonization. In conclusion, MDR and MRS were common in healthy and diseased cats in Saudi Arabia. Thus, an effective antimicrobial stewardship program and further studies using a One Health approach are required to investigate the role of cats as vectors for AMR transmission to humans.
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Rey Pérez, Joaquín, Laura Zálama Rosa, Alfredo García Sánchez, Javier Hermoso de Mendoza Salcedo, Juan Manuel Alonso Rodríguez, Rosario Cerrato Horrillo, Sofía Gabriela Zurita, and María Gil Molino. "Multiple Antimicrobial Resistance in Methicillin-Resistant Staphylococcus sciuri Group Isolates from Wild Ungulates in Spain." Antibiotics 10, no. 8 (July 28, 2021): 920. http://dx.doi.org/10.3390/antibiotics10080920.
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The aim of this study was to investigate the presence of methicillin-resistant Staphylococcus (MRS) strains in non-managed wild ungulates present in a typical Mediterranean forest in Spain. For this purpose, nasal swabs were obtained from 139 animals: 90 wild boar (Sus scrofa), 42 red deer (Cervus elaphus) and 7 fallow deer (Dama dama), which were subsequently pre-enriched in BHI+ NaCl (6.5%) (24 h/37 °C), and then seeded in Columbia blood agar (24 h/37 °C)). The presence of the mecA gene was investigated by PCR, first from the confluent and then from individual colonies. A total of 10 mecA+ colonies were obtained of which only seven showed phenotypic resistance to oxacillin/cefoxitin (methicillin resistance). All MRS strains belonged to the Staphylococcus sciuri group. Methicillin-resistant Staphylococcus aureus (MRSA) was not detected. In addition, a significant number of MRS strains showed resistance to other antimicrobials, mainly β-lactam (7/7), gentamicin (7/7), fusidic acid (6/7) and quinupristin-dalfopristin (6/7), showing an irregular correlation with their coding genes. The genetic profiles grouped the seven strains obtained according to the bacterial species but not in relation to the animal source or the geographical place of origin. The presence of SCCmec type III, common to animals and humans, has been detected in three of the strains obtained. In conclusion, the study reveals that the wild ungulates investigated play a role as potential reservoirs of multi-resistant strains of MRS. Such strains, due to their characteristics, can be easily transferred to other wild or domestic animal species and ultimately to humans through their products.
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Endo, Akihito, and Sanae Okada. "Oenococcus kitaharae sp. nov., a non-acidophilic and non-malolactic-fermenting oenococcus isolated from a composting distilled shochu residue." International Journal of Systematic and Evolutionary Microbiology 56, no. 10 (October 1, 2006): 2345–48. http://dx.doi.org/10.1099/ijs.0.64288-0.
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Six strains of lactic acid bacteria were isolated in Japan from a composting distilled shochu residue. The six isolates grew poorly on MRS agar and slowly in MRS broth. The 16S rRNA gene sequences did not show high levels of similarity to those of the recognized species of lactic acid bacteria, and formed a subcluster within the cluster comprising obligately heterofermentative lactic acid bacteria closely related to Oenococcus oeni. The levels of DNA–DNA relatedness revealed that the isolates belonged to the same taxon and were genetically separate from O. oeni. Furthermore, various phenotypic characteristics such as the optimum pH for growth, malolactic fermentation and resistance to 10 % ethanol revealed that the isolates are distinguishable from O. oeni. On the basis of their phylogenetic and phenotypic characteristics, the isolates represent a novel species, for which the name Oenococcus kitaharae sp. nov. is proposed. The type strain is NRIC 0645T (=JCM 13282T=DSM 17330T).
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Ezzati, Mojgan, Alan Bainbridge, Kevin D. Broad, Go Kawano, Aaron Oliver-Taylor, Eridan Rocha-Ferreira, Daniel Alonso-Alconada, et al. "Immediate remote ischemic postconditioning after hypoxia ischemia in piglets protects cerebral white matter but not grey matter." Journal of Cerebral Blood Flow & Metabolism 36, no. 8 (October 8, 2015): 1396–411. http://dx.doi.org/10.1177/0271678x15608862.
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Remote ischemic postconditioning (RIPostC) is a promising therapeutic intervention whereby brief episodes of ischemia/reperfusion of one organ (limb) mitigate damage in another organ (brain) that has experienced severe hypoxia-ischemia. Our aim was to assess whether RIPostC is protective following cerebral hypoxia-ischemia in a piglet model of neonatal encephalopathy (NE) using magnetic resonance spectroscopy (MRS) biomarkers and immunohistochemistry. After hypoxia-ischemia (HI), 16 Large White female newborn piglets were randomized to: (i) no intervention ( n = 8); (ii) RIPostC – with four, 10-min cycles of bilateral lower limb ischemia/reperfusion immediately after HI ( n = 8). RIPostC reduced the hypoxic-ischemic-induced increase in white matter proton MRS lactate/N acetyl aspartate ( p = 0.005) and increased whole brain phosphorus-31 MRS ATP ( p = 0.039) over the 48 h after HI. Cell death was reduced with RIPostC in the periventricular white matter ( p = 0.03), internal capsule ( p = 0.002) and corpus callosum ( p = 0.021); there was reduced microglial activation in corpus callosum ( p = 0.001) and more surviving oligodendrocytes in corpus callosum ( p = 0.029) and periventricular white matter ( p = 0.001). Changes in gene expression were detected in the white matter at 48 h, including KATP channel and endothelin A receptor. Immediate RIPostC is a potentially safe and promising brain protective therapy for babies with NE with protection in white but not grey matter.
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Nguyen, Doan, Vi Tran, Alireza Shirazian, Cruz Velasco-Gonzalez, and Ifeanyi Iwuchukwu. "Anti-inflammatory genes in PBMCs post-spontaneous intracerebral hemorrhage." Translational Neuroscience 12, no. 1 (January 1, 2021): 58–66. http://dx.doi.org/10.1515/tnsci-2021-0003.
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Abstract Background Neuroinflammation is important in the pathophysiology of spontaneous intracerebral hemorrhage (ICH) and peripheral inflammatory cells play a role in the clinical evolution and outcome. Methodology Blood samples from ICH patients (n = 20) were collected at admission for 5 consecutive days for peripheral blood mononuclear cells (PBMCs). Frozen PBMCs were used for real-time PCR using Taqman probes (NFKB1, SOD1, PPARG, IL10, NFE2L2, and REL) and normalized to GAPDH. Data on hospital length of stay and modified Rankin score (MRS) were collected with 90-day MRS ≤ 3 as favorable outcome. Statistical analysis of clinical characteristics to temporal gene expression from early to delayed timepoints was compared for MRS groups (favorable vs unfavorable) and hematoma volume. Principle findings and results IL10, SOD1, and REL expression were significantly higher at delayed timepoints in PBMCs of ICH patients with favorable outcome. PPARG and REL increased between timepoints in patients with favorable outcome. NFKB1 expression was not sustained, but significantly decreased from higher levels at early onset in patients with unfavorable outcome. IL10 expression showed a negative correlation in patients with high hematoma volume (>30 mL). Conclusions and significance Anti-inflammatory, pro-survival regulators were highly expressed at delayed time points in ICH patients with a favorable outcome, and IL10 expression showed a negative correlation to high hematoma volume.
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Hakumäki, Juhana M., Harish Poptani, Anu-Maaria Sandmair, Seppo Ylä-Herttuala, and Risto A. Kauppinen. "1H MRS detects polyunsaturated fatty acid accumulation during gene therapy of glioma: Implications for the in vivo detection of apoptosis." Nature Medicine 5, no. 11 (November 1999): 1323–27. http://dx.doi.org/10.1038/15279.
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Saqib, Z., G. De Palma, J. Lu, P. Bercik, and S. M. Collins. "A43 β-DEFENSINS AS MARKERS OF INTESTINAL DYSBIOSIS: THE NATURE OF CHANGES IN β-DEFENSINS IS DEPENDENT ON THE PROCESS UNDERLYING THE INDUCTION OF DYSBIOSIS." Journal of the Canadian Association of Gastroenterology 3, Supplement_1 (February 2020): 51–52. http://dx.doi.org/10.1093/jcag/gwz047.042.
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Abstract Background Dysbiosis may be defined as a change in the microbial composition or function that results in altered host function. Defensins are antimicrobial peptides, are part of innate immunity, and are important in host defense and maintaining homeostasis. Dysbiosis is a putative mechanism underlying the expression of many functional GI disorders like Irritable Bowel Syndrome (IBS) for which no biomarkers exist. Previous studies have revealed increased β-defensin (β-Def) levels in IBS patients, most likely due to changes in the microbiota. Aims We examined the hypotheses that: 1) Changes in β-Def are dependent on the manner in which dysbiosis is induced, and that 2) the direction of the change in β-Def depends on how dysbiosis was induced. Methods We used 4 models of experimentally induced dysbiosis to determine changes in fecal β-Def and to characterize the microbiota composition before and during the induction of dysbiosis. We used: 1) an antimicrobial cocktail (AC) in water; 2) a high-fat/ high-sugar diet (HFHSD); 3) a high salt diet (HSD) that we previously showed to induce a pro-inflammatory microbiota; and 4) mild restraint stress (MRS). All studies were performed in C57/BL6 mice except studies using MRS that were performed in NIH Swiss mice. In the AC or dietary studies, we employed a one-week intervention preceded by one-week baseline and recovery periods. In MRS studies, mice comparisons were made between a control and a stressed group. Stool samples were collected every 24 hours and were assayed for fecal β-Def levels analysis by an ELISA and microbial composition by 16S gene profiling. Results Exposure to AC or dietary change, but not MRS, resulted in significant decreases in fecal β-Def. Additionally, bacterial composition and diversity profiles were different in all mice except MRS mice (control vs. MRS males: p=0.414; control vs. MRS females: p=0.96). In contrast, mice exposed to the HSD revealed a significant increase in β-Def during treatment compared to baseline in both males (p=0.025) and females (p= 0.0019). The AC mice showed the largest changes and significant correlations between changes in β-Def levels and bacterial diversity (males: p=0.013, r=0.6; females: p=0.007, r=0.6) and richness (males: p=0.0008, r=0.70; females: p=0.003, r=0.62). However, no significant correlations were found between specific bacteria and β-Def levels in the HFHSD group. Conclusions We conclude that directional changes in fecal β-Def levels are dependent on the manner in which dysbiosis is induced. The use of β-Def as a biomarker requires comparisons with baseline levels obtained during remission in order to identify dysbiosis presence in microbiota-associated chronic GI conditions like IBS. Such an approach will identify patient subgroups that may benefit from microbiota-directed therapies. Funding Agencies CIHR
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Sajjadi, F. S., F. Aghighi, Z. Vahidinia, A. Azami-Tameh, M. Salami, and S. A. Talaei. "Prenatal urban traffic noise exposure impairs spatial learning and memory and reduces glucocorticoid receptor expression in the hippocampus of male rat offspring." Physiology International 107, no. 2 (June 2020): 209–19. http://dx.doi.org/10.1556/2060.2020.00022.
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AbstractIntroductionExposure to noise stress during early life may permanently affect the structure and function of the central nervous system. The aim of this study was to evaluate the effects of prenatal exposure to urban traffic noise on the spatial learning and memory of the rats' offspring and the expression of glucocorticoid receptors (GRs) in their hippocampi.MethodsThree g\roups of pregnant rats were exposed to recorded urban traffic noise for 1, 2 or 4 h/day during the last week of pregnancy. At the age of 45 days, their male offspring were introduced to the Morris water maze (MWM) for assessment of spatial learning and memory. The corticosterone levels were measured in the offspring's sera by radioimmunoassay, and the relative expression of glucocorticoid and mineralocorticoid receptors (MRs) in their hippocampi was evaluated via RT-PCR.ResultsFacing urban traffic noise for 2 and 4 h/day during the third trimester of pregnancy caused the offspring to spend more time and to travel a larger distance than the controls to find the target platform. Analogously, these two groups were inferior to their control counterparts in the probe test. Also, prenatal noise stress elevated the corticosterone concentration in the sera of the rats' offspring and dose-dependently decreased the relative expression of the mRNA of both GRs and MRs in their hippocampi.ConclusionsUrban traffic noise exposure during the last trimester of pregnancy impairs spatial learning and memory of rat offspring and reduces GRs and MRs gene expression in the hippocampus.
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Kim, C. G., K. M. Barnhart, and M. Sheffery. "Purification of multiple erythroid cell proteins that bind the promoter of the alpha-globin gene." Molecular and Cellular Biology 8, no. 10 (October 1988): 4270–81. http://dx.doi.org/10.1128/mcb.8.10.4270.
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Three erythroid cell factors that bind the murine alpha-globin promoter were enriched more than 1,000-fold by conventional and DNA sequence affinity chromatography. Visualization of enriched polypeptides revealed simple patterns suggesting that each binding activity was purified. Two of the purified proteins, alpha-CP1 and alpha-CP2, have been shown previously to interact with distinct binding sites that overlap in the alpha-globin CCAAT box. Affinity purification of alpha-CP1 revealed seven polypeptides with Mrs raging from 27,000 to 38,000. In contrast, purified alpha-CP2 was made up of a polypeptide doublet with Mrs of 64,000 and 66,000. The third purified binding activity, alpha-IRP, interacted with sequences that formed an inverted repeat (IR) between the alpha-globin CCAAT and TATAA boxes. Affinity-purified alpha-IRP was made up of a single polypeptide with an Mr of 85,000. We confirmed that the purified polypeptides corresponded to alpha-CP1-, alpha-CP2-, and alpha-IRP-binding activities by UV cross-linking experiments (alpha-CP2 and alpha-IRP) or by renaturation of binding activity after elution of polypeptides from sodium dodecyl sulfate-polyacrylamide gels (alpha-CP1 and alpha-CP2). The apparent complexity of the polypeptides accounting for alpha-CP1 binding activity prompted a further physical characterization of this factor. Sedimentation of affinity-purified alpha-CP1 in glycerol gradients containing 100 mM KCl showed that all seven polypeptides migrated as a complex that cosedimented with alpha-CP1-binding activity. In contrast, when sedimented in glycerol gradients containing 500 mM KCl, alpha-CP1 dissociated into at least two components. Under these conditions, alpha-CP1-binding activity was reduced or lost. Activity was reconstituted, however, by combining fractions that were enriched in the two components. These results were confirmed by experiments in which we showed that alpha-CP1-binding activity can be recovered only by combining distinct sets of polypeptides that were isolated and renatured from sodium dodecyl sulfate-polyacrylamide gels. Our results suggest that the seven polypeptides visualized after affinity purification of alpha-CP1 interact to form a heterotypic complex (or set of complexes) required for alpha-CP1-binding activity.
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Kim, C. G., K. M. Barnhart, and M. Sheffery. "Purification of multiple erythroid cell proteins that bind the promoter of the alpha-globin gene." Molecular and Cellular Biology 8, no. 10 (October 1988): 4270–81. http://dx.doi.org/10.1128/mcb.8.10.4270-4281.1988.
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Three erythroid cell factors that bind the murine alpha-globin promoter were enriched more than 1,000-fold by conventional and DNA sequence affinity chromatography. Visualization of enriched polypeptides revealed simple patterns suggesting that each binding activity was purified. Two of the purified proteins, alpha-CP1 and alpha-CP2, have been shown previously to interact with distinct binding sites that overlap in the alpha-globin CCAAT box. Affinity purification of alpha-CP1 revealed seven polypeptides with Mrs raging from 27,000 to 38,000. In contrast, purified alpha-CP2 was made up of a polypeptide doublet with Mrs of 64,000 and 66,000. The third purified binding activity, alpha-IRP, interacted with sequences that formed an inverted repeat (IR) between the alpha-globin CCAAT and TATAA boxes. Affinity-purified alpha-IRP was made up of a single polypeptide with an Mr of 85,000. We confirmed that the purified polypeptides corresponded to alpha-CP1-, alpha-CP2-, and alpha-IRP-binding activities by UV cross-linking experiments (alpha-CP2 and alpha-IRP) or by renaturation of binding activity after elution of polypeptides from sodium dodecyl sulfate-polyacrylamide gels (alpha-CP1 and alpha-CP2). The apparent complexity of the polypeptides accounting for alpha-CP1 binding activity prompted a further physical characterization of this factor. Sedimentation of affinity-purified alpha-CP1 in glycerol gradients containing 100 mM KCl showed that all seven polypeptides migrated as a complex that cosedimented with alpha-CP1-binding activity. In contrast, when sedimented in glycerol gradients containing 500 mM KCl, alpha-CP1 dissociated into at least two components. Under these conditions, alpha-CP1-binding activity was reduced or lost. Activity was reconstituted, however, by combining fractions that were enriched in the two components. These results were confirmed by experiments in which we showed that alpha-CP1-binding activity can be recovered only by combining distinct sets of polypeptides that were isolated and renatured from sodium dodecyl sulfate-polyacrylamide gels. Our results suggest that the seven polypeptides visualized after affinity purification of alpha-CP1 interact to form a heterotypic complex (or set of complexes) required for alpha-CP1-binding activity.
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Pramanick, Rinku, Shraddha Parab, Niranjan Mayadeo, Himangi Warke, and Clara Aranha. "Cross sectional analysis of vaginal Lactobacillus in asymptomatic women of reproductive age in Mumbai, India." Journal of Infection in Developing Countries 12, no. 12 (December 31, 2018): 1096–104. http://dx.doi.org/10.3855/jidc.10154.
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Introduction: Lactobacillus dominated vaginal microenvironment is associated with lower risk of genital infections. Numerous studies have reported geographic and ethnic variations in vaginal microbiome structure between healthy individuals from different race and ethnicity. India has a great diversity, so it is intriguing to find out if such divergences exist in vaginal lactobacilli. The present study aimed to investigate predominant Lactobacillus species in vaginas of healthy Indian women and screen isolates for lactic acid and H2O2 production.
Methodology: 203 premenopausal women asymptomatic for any vaginal complaints were recruited. The lactobacilli isolates on MRS agar were identified by Multiplex-PCR and 16sRNA gene sequencing. RAPD was used to differentiate strains of same species. H2O2 and lactic acid was evaluated on TMB-HRP MRS agar and BCP-MRS agar respectively.
Results: Lactobacilli were recovered from 107/109 (98.2%) women with normal microflora. L. iners 64.7% (68), L. crispatus 26.7% (28), L. reuteri 21.9% (23), L. jensenii 16.2% (17) and L. gasseri 15.2% (16) were the most frequently occurring vaginal lactobacilli in normal women. The vaginal microflora was dominated by either by a single (80%, n = 84) or a combination (20%, n = 21) of Lactobacillus species. Though most frequently identified, L. iners, coexisted only with other Lactobacillus species. All isolates were acid producers but H2O2 was produced by 94.2% isolates.
Conclusions: Our study reports prevalent vaginal lactobacilli which could be explored as probiotics. Presence of heterogeneous Lactobacillus population highlights the cumulative effects of different lactobacilli maintaining vaginal health. Contrasting observations about L. iners reiterates its puzzling role in vaginal immunity, advocating further research.
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Gaston, Sandra M., Jonathan G. Rogg, Dang Vu, Jung M. Lee, Dana L. Goldner, Elizabeth M. Genega, Robert E. Lenkinski, and William C. De Wolf. "839: Gene Expression Profiles that Underlie the Biological Events Visualized by Magnetic Resonance Spectra (MRS) of Human Prostate Cancer: Choline Kinase." Journal of Urology 171, no. 4S (April 2004): 221–22. http://dx.doi.org/10.1016/s0022-5347(18)38088-1.
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Slingo, Mary, Mark Cole, Carolyn Carr, Mary K. Curtis, Michael Dodd, Lucia Giles, Lisa C. Heather, Damian Tyler, Kieran Clarke, and Peter A. Robbins. "The von Hippel-Lindau Chuvash mutation in mice alters cardiac substrate and high-energy phosphate metabolism." American Journal of Physiology-Heart and Circulatory Physiology 311, no. 3 (September 1, 2016): H759—H767. http://dx.doi.org/10.1152/ajpheart.00912.2015.
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Hypoxia-inducible factor (HIF) appears to function as a global master regulator of cellular and systemic responses to hypoxia. HIF pathway manipulation is of therapeutic interest; however, global systemic upregulation of HIF may have as yet unknown effects on multiple processes. We used a mouse model of Chuvash polycythemia (CP), a rare genetic disorder that modestly increases expression of HIF target genes in normoxia, to understand what these effects might be within the heart. An integrated in and ex vivo approach was employed. Compared with wild-type controls, CP mice had evidence (using in vivo magnetic resonance imaging) of pulmonary hypertension, right ventricular hypertrophy, and increased left ventricular ejection fraction. Glycolytic flux (measured using [3H]glucose) in the isolated contracting perfused CP heart was 1.8-fold higher. Net lactate efflux was 1.5-fold higher. Furthermore, in vivo 13C-magnetic resonance spectroscopy (MRS) of hyperpolarized [13C1]pyruvate revealed a twofold increase in real-time flux through lactate dehydrogenase in the CP hearts and a 1.6-fold increase through pyruvate dehydrogenase. 31P-MRS of perfused CP hearts under increased workload (isoproterenol infusion) demonstrated increased depletion of phosphocreatine relative to ATP. Intriguingly, no changes in cardiac gene expression were detected. In summary, a modest systemic dysregulation of the HIF pathway resulted in clear alterations in cardiac metabolism and energetics. However, in contrast to studies generating high HIF levels within the heart, the CP mice showed neither the predicted changes in gene expression nor any degree of LV impairment. We conclude that the effects of manipulating HIF on the heart are dose dependent.
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Peric, Predrag, Goran Pavlicevic, Jelena Ostojic, Dejan Kostic, Sanja Nikolajevic, Gordana Supic, Zvonko Magic, and Sanja Radovinovic-Tasic. "Synchronous malignant multicentric cerebral glioma with atypical neuroradiological presentation and comparatively long survival: Case report and literature review." Vojnosanitetski pregled 75, no. 4 (2018): 414–21. http://dx.doi.org/10.2298/vsp160605238p.
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Introduction. Synchronous multicentric cerebral gliomas are uncommon brain tumors, mostly malignant, with unknown pathogenesis, unfavorable prognosis and still controversial management. Preoperative differentiation from other multiple brain pathologies by conventional magnetic resonance imaging (MRI) is often difficult, but supplemental use of advanced magnetic resonance techniques should allow the tumor biology to be predicted and an appropriate treatment strategy planned. Case report. We reported a 59-yearold man with double synchronous multicentric cerebral lesions, which had initial MRI and diffusion-weighted imaging presentation as left parietal metastasis and ipsilateral amygdalo- hippocampal low-grade glioma. However, magnetic resonance spectroscopy (MRS) of both lesions showed different metabolite profiles of malignant glioma. En bloc resection of the easily accessible parietal lesion revealed glioblastoma with methylated O6-methylguanine-DNA methyltransferase (MGMT) gene promoter. Subsequently, the patient was treated with temozolomide (TMZ)-based chemoradiation according to Stupp?s protocol, with continuous standard (5/28) adjuvant TMZ in 12 courses. Despite prolonged stabilization of the disease with good life-quality during treatment, the patient died 19 months after diagnosis. The time to tumor progression estimated by MRI was 17 months. Conclusion. MRS significantly improved the differential diagnostic accuracy of conventional MRI in our patient. In accordance with reviewed literature data, the younger age, good initial performance status and methylated MGMT gene promoter were all favorable predictors of longer survival in the reported case. Resection of at least one easily accessible tumor lesion, followed by TMZ-based chemoradiation, with continuous adjuvant TMZ in more than 6 standard courses, seems currently to be the most beneficial therapeutic option for such cases.
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Alexander, Sheila, Samuel Poloyac, Leslie Hoffman, Matthew Gallek, Dianxu Ren, Jeffrey Balzer, Amin Kassam, and Yvette Conley. "Endothelial Nitric Oxide Synthase Tagging Single Nucleotide Polymorphisms and Recovery From Aneurysmal Subarachnoid Hemorrhage." Biological Research For Nursing 11, no. 1 (May 5, 2009): 42–52. http://dx.doi.org/10.1177/1099800409334751.
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Aneurysmal subarachnoid hemorrhage (SAH) is a hemorrhagic stroke subtype with a poor recovery profile. Cerebral vasospasm (CV), a narrowing of the cerebral vasculature, significantly contributes to the poor recovery profile. Variation in the endothelial nitric oxide (NO) synthase (eNOS) gene has been implicated in CV and outcome after SAH. The purpose of this project was to explore the potential association between three eNOS tagging single nucleotide polymorphisms (SNPs) and recovery from SAH. We included 195 participants with a diagnosis of SAH and DNA and 6-month outcome data available but without preexisting neurologic disease/deficit. Genotyping was performed using an ABI Prism 7000 Sequence Detection System and TaqMan assays. CV was verified by cerebral angiogram independently read by a neurosurgeon on 118 participants. Modified Rankin Scores (MRS) and Glasgow Outcome Scale (GOS) scores were collected 6 months posthemorrhage. Data were analyzed using descriptive statistics, analysis of variance (ANOVA) and chi-square analysis as appropriate. The sample was primarily female (n = 147; 75.4%) and White (n = 178; 91.3%) with a mean age of 54.6 years. Of the participants with CV data, 56 (47.5%) developed CV within 14 days of SAH. None of the SNPs individually were associated with CV presence; however, a combination of the three variant SNPs was significantly associated with CV (p = .017). Only one SNP (rs1799983, variant allele) was associated with worse 6-month GOS scores (p < .001) and MRS (p < .001). These data indicate that the eNOS gene plays a role in the response to SAH, which may be explained by an influence on CV.
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Højberg, Ole, Nuria Canibe, Hanne Damgaard Poulsen, Mette Skou Hedemann, and Bent Borg Jensen. "Influence of Dietary Zinc Oxide and Copper Sulfate on the Gastrointestinal Ecosystem in Newly Weaned Piglets." Applied and Environmental Microbiology 71, no. 5 (May 2005): 2267–77. http://dx.doi.org/10.1128/aem.71.5.2267-2277.2005.
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ABSTRACT Dietary doses of 2,500 ppm ZnO-Zn reduced bacterial activity (ATP accumulation) in digesta from the gastrointestinal tracts of newly weaned piglets compared to that in animals receiving 100 ppm ZnO-Zn. The amounts of lactic acid bacteria (MRS counts) and lactobacilli (Rogosa counts) were reduced, whereas coliforms (MacConkey counts) and enterococci (Slanetz counts, red colonies) were more numerous in animals receiving the high ZnO dose. Based on 16S rRNA gene sequencing, the colonies on MRS were dominated by three phylotypes, tentatively identified as Lactobacillus amylovorus (OTU171), Lactobacillus reuteri (OTU173), and Streptococcus alactolyticus (OTU180). The colonies on Rogosa plates were dominated by the two Lactobacillus phylotypes only. Terminal restriction fragment length polymorphism analysis supported the observations of three phylotypes of lactic acid bacteria dominating in piglets receiving the low ZnO dose and of coliforms and enterococci dominating in piglets receiving the high ZnO dose. Dietary doses of 175 ppm CuSO4-Cu also reduced MRS and Rogosa counts of stomach contents, but for these animals, the numbers of coliforms were reduced in the cecum and the colon. The influence of ZnO on the gastrointestinal microbiota resembles the working mechanism suggested for some growth-promoting antibiotics, namely, the suppression of gram-positive commensals rather than potentially pathogenic gram-negative organisms. Reduced fermentation of digestible nutrients in the proximal part of the gastrointestinal tract may render more energy available for the host animal and contribute to the growth-promoting effect of high dietary ZnO doses. Dietary CuSO4 inhibited the coliforms and thus potential pathogens as well, but overall the observed effect of CuSO4 was limited compared to that of ZnO.
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O'Flaherty, Sarah J., and Todd R. Klaenhammer. "Functional and phenotypic characterization of a protein from Lactobacillus acidophilus involved in cell morphology, stress tolerance and adherence to intestinal cells." Microbiology 156, no. 11 (November 1, 2010): 3360–67. http://dx.doi.org/10.1099/mic.0.043158-0.
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Structural components of the cell surface have an impact on some of the beneficial attributes of probiotic bacteria. In silico analysis of the L. acidophilus NCFM genome sequence revealed the presence of a putative cell surface protein that was predicted to be a myosin cross-reactive antigen (MCRA). As MCRAs are conserved among many probiotic bacteria, we used the upp-based counterselective gene replacement system, designed recently for use in L. acidophilus, to determine the functional role of this gene (LBA649) in L. acidophilus NCFM. Phenotypic assays were undertaken with the parent strain (NCK1909) and deletion mutant (NCK2015) to assign a function for this gene. The growth of NCK2015 (ΔLBA649) was reduced in the presence of lactate, acetate, porcine bile and salt. Adhesion of NCK2015 to Caco-2 cells was substantially reduced for both stationary-phase (∼45 % reduction) and exponential-phase cells (∼50 % reduction). Analysis of NCK2015 by scanning electron microscopy revealed a longer cell morphology after growth in MRS broth compared to NCK1909. These results indicate a role for LBA649 in stress tolerance, cell wall division and adherence to Caco-2 cells.
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Ferreira, A. B., M. N. V. de Oliveira, F. S. Freitas, A. D. Paiva, P. Alfenas-Zerbini, D. F. da Silva, M. V. de Queiroz, A. C. Borges, and C. A. de Moraes. "Involvement of the ornithine decarboxylase gene in acid stress response in probiotic Lactobacillus delbrueckii UFV H2b20." Beneficial Microbes 6, no. 5 (October 15, 2015): 719–25. http://dx.doi.org/10.3920/bm2014.0122.
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Amino acid decarboxylation is important for the maintenance of intracellular pH under acid stress. This study aims to carry out phylogenetic and expression analysis by real-time PCR of two genes that encode proteins involved in ornithine decarboxylation in Lactobacillus delbrueckii UFV H2b20 exposed to acid stress. Sequencing and phylogeny analysis of genes encoding ornithine decarboxylase and amino acid permease in L. delbrueckii UFV H2b20 showed their high sequence identity (99%) and grouping with those of L. delbrueckii subsp. bulgaricus ATCC 11842. Exposure of L. delbrueckii UFV H2b20 cells in MRS pH 3.5 for 30 and 60 min caused a significant increase in expression of the gene encoding ornithine decarboxylase (up to 8.1 times higher when compared to the control treatment). Increased expression of the ornithine decarboxylase gene demonstrates its involvement in acid stress response in L. delbrueckii UFV H2b20, evidencing that the protein encoded by that gene could be involved in intracellular pH regulation. The results obtained show ornithine decarboxylation as a possible mechanism of adaptation to an acidic environmental condition, a desirable and necessary characteristic for probiotic cultures and certainly important to the survival and persistence of the L. delbrueckii UFV H2b20 in the human gastrointestinal tract.
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Casasnovas, Carlos, Edgard Verdura, Valentina Vélez, Agatha Schlüter, Albert Pons-Escoda, Christian Homedes, Montserrat Ruiz, Stéphane Fourcade, Nathalie Launay, and Aurora Pujol. "A novel mutation in the GFAP gene expands the phenotype of Alexander disease." Journal of Medical Genetics 56, no. 12 (April 19, 2019): 846–49. http://dx.doi.org/10.1136/jmedgenet-2018-105959.
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BackgroundAlexander disease, an autosomal dominant leukodystrophy, is caused by missense mutations in GFAP. Although mostly diagnosed in children, associated with severe leukoencephalopathy, milder adult forms also exist.MethodsA family affected by adult-onset spastic paraplegia underwent neurological examination and cerebral MRI. Two patients were sequenced by whole exome sequencing (WES). A candidate variant was functionally tested in an astrocytoma cell line.ResultsThe novel variant in GFAP (Glial Fibrillary Acidic Protein) N-terminal head domain (p.Gly18Val) cosegregated in multiple relatives (LOD score: 2.7). All patients, even those with the mildest forms, showed characteristic signal changes or atrophy in the brainstem and spinal cord MRIs, and abnormal MRS. In vitro, this variant did not cause significant protein aggregation, in contrast to most Alexander disease mutations characterised so far. However, cell area analysis showed larger size, a feature previously described in patients and mouse models.ConclusionWe suggest that this variant causes variable expressivity and an attenuated phenotype of Alexander disease type II, probably associated with alternative pathogenic mechanisms, that is, astrocyte enlargement. GFAP analysis should be considered in adult-onset neurological presentations with pyramidal and bulbar symptoms, in particular when characteristic findings, such as the tadpole sign, are present in MRI. WES is a powerful tool to diagnose atypical cases.
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Toldo, Irene, Diego Cecchin, Stefano Sartori, Milena Calderone, Rodica Mardari, Francesca Cattelan, Anna Maria Laverda, Paola Drigo, and Pier Antonio Battistella. "Multimodal neuroimaging in a child with sporadic hemiplegic migraine: A contribution to understanding pathogenesis." Cephalalgia 31, no. 6 (December 20, 2010): 751–56. http://dx.doi.org/10.1177/0333102410392068.
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Background: Hemiplegic migraine (HM) is a rare variety of migraine with aura, characterized by motor deficits during the aura, often beginning in childhood. The hemiplegic attacks can be severe and prolonged but the prognosis is usually good. Data on neuroimaging, including diffusion-weighted imaging (DWI) and spectroscopy, during prolonged attacks of HM are quite limited, particularly in children. Case: An eight-year-old female had a prolonged attack of sporadic HM characterized by right-sided hemiplegia, global aphasia, fever and impairment of consciousness. MRI nine hours after hemiplegia onset was negative, while the following MRI scans (days 4 and 11) documented a progressive increase in cortical swelling in the left hemisphere with mild hyperintensity on DWI and mild reduction of apparent diffusion coefficient values. Proton MRI spectroscopy (MRS) (day 15) showed a decrease in the N-acetylaspartate/creatine ratio in the left hemisphere. 99mTc-ECD single-photon emission tomography (SPET) (day 27) showed marked left hemispheric hypoperfusion. The patient recovered completely after 40 days and neuroimaging follow-up (MRI and SPET) after six months was normal. The patient carried a missense mutation of the ATP1A2 gene. Conclusion: Multimodal neuroimaging (MRI, DWI, MRS, SPET) in a prolonged HM attack supports evidence for a primary neuronal dysfunction.
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De Filippis, Bianca, Mattia Musto, Luisa Altabella, Emilia Romano, Rossella Canese, and Giovanni Laviola. "Deficient Purposeful Use of Forepaws in Female Mice Modelling Rett Syndrome." Neural Plasticity 2015 (2015): 1–13. http://dx.doi.org/10.1155/2015/326184.
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Rett syndrome (RTT) is a rare neurodevelopmental disorder, characterized by severe behavioural and physiological symptoms. Mutations in the methyl CpG binding protein 2 gene (MECP2) cause more than 95% of classic cases. Motor abnormalities represent a significant part of the spectrum of RTT symptoms. In the present study we investigated motor coordination and fine motor skill domains in MeCP2-308 female mice, a validated RTT model. This was complemented by thein vivomagnetic resonance spectroscopy (MRS) analysis of metabolic profile in behaviourally relevant brain areas. MeCP2-308 heterozygous female mice (Het, 10-12 months of age) were impaired in tasks validated for the assessment of purposeful and coordinated forepaw use (Morag testandCapellini handling task). A fine-grain analysis of spontaneous behaviour in the home-cage also revealed an abnormal handling pattern when interacting with the nesting material, reduced motivation to explore the environment, and increased time devoted to feeding in Het mice. The brain MRS evaluation highlighted decreased levels of bioenergetic metabolites in the striatal area in Het mice compared to controls. Present results confirm behavioural and brain alterations previously reported in MeCP2-308 males and identify novel endpoints on which the efficacy of innovative therapeutic strategies for RTT may be tested.