Investigation of the effect of bcl-2 on hybridoma cell survival

Hybridoma cells are widely used to produce large quantities of monoclonal antibodies for use as medicines and diagnostics. Bcl-2 is an oncogene that has been shown to have an anti-apoptotic effect in many cell lines including hybridomas. This study investigates the effect of various levels of bcl-2 expression on hybridoma cell grown and antibody productivity in both batch and perfusion culture.;H166 hybridomas were transfected with a plasmid containing a bcl-2 insert and stable clones were selected. The cells were grown in spinner flasks in batch mode for twelve days and it was confirmed that expression at high levels allow for significant protection from apoptosis. Protection from apoptosis first becomes evident at 2--5 units/cell of BCL-2 protein content, where one unit of expression is the amount of BCL-2 content per HL-60 cell taken at 48 hr of batch culture. Further protection from apoptosis is obtained as the level of bcl-2 expression increases to 27 units. Although there is a significant increase in the viability during the decline phase of cell culture, there is no improvement in the antibody productivity. The antibody concentration levels off after 7 days of culture. Bcl-2 expression had no effect on nutrient consumption or cell cycle progression.;Growth of the cells in long-term perfusion culture did not result in an increase in viable cell density, viability or antibody productivity. At low cell bleed rates the vector only control grew better than any of the clones tested. At high cell bleed rates, where cell death due to apoptosis was expected to be low, there was an increase in cell density for the highest expressing clone. This was not due to prevention of apoptosis and was likely due to culture variations. The extent of apoptosis remained low in all the cultures studied. These results suggest that induction of bcl-2 expression is not an effective method to increase the antibody productivity of H166 hybridoma cells in batch or perfusion culture

Standard chemoradiation versus intensity-modulated chemoradiation: a quality of life assessment in oropharyngeal cancer patients

This study is based on the context that many patients with advanced oropharyngeal carcinoma are being treated with primary chemoradiation. The aims of this study are to identify differences in quality of life (QOL) between patients with advanced oropharyngeal cancer following traditional chemoradiation versus chemotherapy with intensity-modulated radiation therapy (CIMRT). This research is designed on a cohort study from an academic tertiary referral center. Fifty patients were identified from an institutional database of patients who had undergone primary chemotherapy and radiation (traditional or IMRT) for advanced oropharyngeal carcinoma. Patients responded via mail using the University of Washington quality of life instrument version 4. Statistical analysis of data was performed using Chi-square and Wilcoxon tests. The results comprise the responses of 17 CRT (57%) and 14 CIMRT (70%) patients. The patients completed the survey between 9 and 44 months following end of treatment. When adjusted for tumor stage and time since treatment, CIMRT patients reported improved appearance (p = 0.05), chewing (p = 0.02), and mood (p = 0.01). There was a trend toward significance for improved activity (p = 0.07), recreation (p = 0.07), and anxiety (p = 0.08). There were no differences between the two groups for saliva, taste, shoulder function, speech, and swallowing. But there was a trend for significance for improved overall QOL in patients who had undergone CIMRT (p = 0.06). In conclusion, CIMRT results in improved QOL for some domains but surprisingly not for swallowing or saliva. Patients undergoing CIMRT also report slightly better QOL overall when compared to patients receiving more traditional forms of radiation therapy

Prognostication and contemporary management of clinically isolated syndrome

Clinically isolated syndrome (CIS) patients present with a single attack of inflammatory demyelination of the central nervous system. Recent advances in multiple sclerosis (MS) diagnostic criteria have expanded the number of CIS patients eligible for a diagnosis of MS at the onset of the disease, shrinking the prevalence of CIS. MS treatment options are rapidly expanding, which is driving the need to recognise MS at its earliest stages. In CIS patients, finding typical MS white matter lesions on the patient's MRI scan remains the most influential prognostic investigation for predicting subsequent diagnosis with MS. Additional imaging, cerebrospinal fluid and serum testing, information from the clinical history and genetic testing also contribute. For those subsequently diagnosed with MS, there is a wide spectrum of long-term clinical outcomes. Detailed assessment at the point of presentation with CIS provides fewer clues to calculate a personalised risk of long-term severe disability. Clinicians should select suitable CIS cases for steroid treatment to speed neurological recovery. Unfortunately, there are still no neuroprotection or remyelination strategies available. The use of MS disease modifying therapy for CIS varies among clinicians and national guidelines, suggesting a lack of robust evidence to guide practice. Clinicians should focus on confirming MS speedily and accurately with appropriate investigations. Diagnosis with CIS provides an opportune moment to promote a healthy lifestyle, in particular smoking cessation. Patients also need to understand the link between CIS and MS. This review provides clinicians an update on the contemporary evidence guiding prognostication and management of CIS

Towards a Better Understanding of Rural Homelessness: An Examination of Housing Crisis in a Small, Rural Minnesota Community

This report compiles the work done during the Rural Housing Policy course at the University of Minnesota Morris by the students and their instructor, Professor Greg Thorson. the class reviewed the literature on urban and rural homelessness, interviewed local providers of social service programs, developed a survey to be administered at regional homeless shelters, wrote the Institutional Review Board (IRB) proposal to authorize the administration of the survey, administered the survey, and analyzed the results.https://digitalcommons.morris.umn.edu/cst/1009/thumbnail.jp

Fibroblast and Lymphoblast Gene Expression Profiles in Schizophrenia: Are Non-Neural Cells Informative?

Lymphoblastoid cell lines (LCLs) and fibroblasts provide conveniently derived non-neuronal samples in which to investigate the aetiology of schizophrenia (SZ) using gene expression profiling. This assumes that heritable mechanisms associated with risk of SZ have systemic effects and result in changes to gene expression in all tissues. The broad aim of this and other similar studies is that comparison of the transcriptomes of non-neuronal tissues from SZ patients and healthy controls may identify gene/pathway dysregulation underpinning the neurobiological defects associated with SZ. Using microarrays consisting of 18,664 probes we compared gene expression profiles of LCLs from SZ cases and healthy controls. To identify robust associations with SZ that were not patient or tissue specific, we also examined fibroblasts from an independent series of SZ cases and controls using the same microarrays. In both tissue types ANOVA analysis returned approximately the number of differentially expressed genes expected by chance. No genes were significantly differentially expressed in either tissue when corrected for multiple testing. Even using relaxed parameters (p≤0.05, without multiple testing correction) there were still no differentially expressed genes that also displayed ≥2-fold change between the groups of SZ cases and controls common to both LCLs and fibroblasts. We conclude that despite encouraging data from previous microarray studies assessing non-neural tissues, the lack of a convergent set of differentially expressed genes associated with SZ using fibroblasts and LCLs indicates the utility of non-neuronal tissues for detection of gene expression differences and/or pathways associated with SZ remains to be demonstrated

Student Recital (December 6, 2013)

Concert Étude, Op. 49 / Alexander Goedicke James Sheehan, trumpet Andante / Marco Bordogni Kevin Torres, trombone Sonata No. 9 in D Major, K. 311 / Wolfgang Amadeus Mozart Allegro Jiaying Zhu, piano Etude 17 / Fernando Sor Tim Prosser, guitar Sonata, Op. 61 / Joaquín Turina Andante Brian Strange, guitar El Decameron negro / Leo Brouwer La Fuite des Amants par la Vallée Echos Mark Gavin, guitar Sonata No. 3 for Lute in C Major, BWV 1005 / Johann Sebastian Bach Adagio Ian Timpany, guitar Estudio en Sol Menor / Agustin Barrios Mangoré Christopher Bosch, guitar Capricho Arabe / Francisco Tarrega Dylan Mowry, guitarhttps://vc.bridgew.edu/student_concerts/1050/thumbnail.jp

A Rare Functional Noncoding Variant at the GWAS-Implicated MIR137/MIR2682 Locus Might Confer Risk to Schizophrenia and Bipolar Disorder

Schizophrenia (SZ) genome-wide association studies (GWASs) have identified common risk variants in >100 susceptibility loci; however, the contribution of rare variants at these loci remains largely unexplored. One of the strongly associated loci spans MIR137 (miR137) and MIR2682 (miR2682), two microRNA genes important for neuronal function. We sequenced ∼6.9 kb MIR137/MIR2682 and upstream regulatory sequences in 2,610 SZ cases and 2,611 controls of European ancestry. We identified 133 rare variants with minor allele frequency (MAF) <0.5%. The rare variant burden in promoters and enhancers, but not insulators, was associated with SZ (p = 0.021 for MAF < 0.5%, p = 0.003 for MAF < 0.1%). A rare enhancer SNP, 1:g.98515539A>T, presented exclusively in 11 SZ cases (nominal p = 4.8 × 10−4). We further identified its risk allele T in 2 of 2,434 additional SZ cases, 11 of 4,339 bipolar (BP) cases, and 3 of 3,572 SZ/BP study controls and 1,688 population controls; yielding combined p values of 0.0007, 0.0013, and 0.0001 for SZ, BP, and SZ/BP, respectively. The risk allele T of 1:g.98515539A>T reduced enhancer activity of its flanking sequence by >50% in human neuroblastoma cells, predicting lower expression of MIR137/MIR2682. Both empirical and computational analyses showed weaker transcription factor (YY1) binding by the risk allele. Chromatin conformation capture (3C) assay further indicated that 1:g.98515539A>T influenced MIR137/MIR2682, but not the nearby DPYD or LOC729987. Our results suggest that rare noncoding risk variants are associated with SZ and BP at MIR137/MIR2682 locus, with risk alleles decreasing MIR137/MIR2682 expression

Beyond the Global Brain Differences:Intraindividual Variability Differences in 1q21.1 Distal and 15q11.2 BP1-BP2 Deletion Carriers

BACKGROUND: Carriers of the 1q21.1 distal and 15q11.2 BP1-BP2 copy number variants exhibit regional and globalbrain differences compared with noncarriers. However, interpreting regional differences is challenging if a globaldifference drives the regional brain differences. Intraindividual variability measures can be used to test for regionaldifferences beyond global differences in brain structure.METHODS: Magnetic resonance imaging data were used to obtain regional brain values for 1q21.1 distal deletion (n =30) and duplication (n = 27) and 15q11.2 BP1-BP2 deletion (n = 170) and duplication (n = 243) carriers and matchednoncarriers (n = 2350). Regional intra-deviation scores, i.e., the standardized difference between an individual’sregional difference and global difference, were used to test for regional differences that diverge from the globaldifference.RESULTS: For the 1q21.1 distal deletion carriers, cortical surface area for regions in the medial visual cortex, posterior cingulate, and temporal pole differed less and regions in the prefrontal and superior temporal cortex differedmore than the global difference in cortical surface area. For the 15q11.2 BP1-BP2 deletion carriers, cortical thicknessin regions in the medial visual cortex, auditory cortex, and temporal pole differed less and the prefrontal andsomatosensory cortex differed more than the global difference in cortical thickness.CONCLUSIONS: We find evidence for regional effects beyond differences in global brain measures in 1q21.1 distaland 15q11.2 BP1-BP2 copy number variants. The results provide new insight into brain profiling of the 1q21.1 distaland 15q11.2 BP1-BP2 copy number variants, with the potential to increase understanding of the mechanismsinvolved in altered neurodevelopment

Cortical brain abnormalities in 4474 individuals with schizophrenia and 5098 control subjects via the enhancing neuro Imaging genetics through meta analysis (ENIGMA) Consortium

BACKGROUND: The profile of cortical neuroanatomical abnormalities in schizophrenia is not fully understood, despite hundreds of published structural brain imaging studies. This study presents the first meta-analysis of cortical thickness and surface area abnormalities in schizophrenia conducted by the ENIGMA (Enhancing Neuro Imaging Genetics through Meta Analysis) Schizophrenia Working Group. METHODS: The study included data from 4474 individuals with schizophrenia (mean age, 32.3 years; range, 11-78 years; 66% male) and 5098 healthy volunteers (mean age, 32.8 years; range, 10-87 years; 53% male) assessed with standardized methods at 39 centers worldwide. RESULTS: Compared with healthy volunteers, individuals with schizophrenia have widespread thinner cortex (left/right hemisphere: Cohen's d = -0.530/-0.516) and smaller surface area (left/right hemisphere: Cohen's d = -0.251/-0.254), with the largest effect sizes for both in frontal and temporal lobe regions. Regional group differences in cortical thickness remained significant when statistically controlling for global cortical thickness, suggesting regional specificity. In contrast, effects for cortical surface area appear global. Case-control, negative, cortical thickness effect sizes were two to three times larger in individuals receiving antipsychotic medication relative to unmedicated individuals. Negative correlations between age and bilateral temporal pole thickness were stronger in individuals with schizophrenia than in healthy volunteers. Regional cortical thickness showed significant negative correlations with normalized medication dose, symptom severity, and duration of illness and positive correlations with age at onset. CONCLUSIONS: The findings indicate that the ENIGMA meta-analysis approach can achieve robust findings in clinical neuroscience studies; also, medication effects should be taken into account in future genetic association studies of cortical thickness in schizophrenia

The genetic architecture of the human cerebral cortex

The cerebral cortex underlies our complex cognitive capabilities, yet little is known about the specific genetic loci that influence human cortical structure. To identify genetic variants that affect cortical structure, we conducted a genome-wide association meta-analysis of brain magnetic resonance imaging data from 51,665 individuals. We analyzed the surface area and average thickness of the whole cortex and 34 regions with known functional specializations. We identified 199 significant loci and found significant enrichment for loci influencing total surface area within regulatory elements that are active during prenatal cortical development, supporting the radial unit hypothesis. Loci that affect regional surface area cluster near genes in Wnt signaling pathways, which influence progenitor expansion and areal identity. Variation in cortical structure is genetically correlated with cognitive function, Parkinson's disease, insomnia, depression, neuroticism, and attention deficit hyperactivity disorder