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OBJECTIVES: To evaluate the immunohistochemical expression of Epstein-Barr virus (EBV) in broad spectrum histological subtypes of oral squamous cell carcinoma (OSCC) and to determine the relationship of EBV with clinicopathological parameters of OSCC. METHODS: This cross-sectional study included 150 clinically diagnosed OSCC cases from the outpatient of Ziauddin University Hospital from March, 2017 to October, 2018. These were confirmed on histological examination and categorized into conventional squamous cell carcinoma (SCC) and rare variants. Conventional SCC was subcategorized into keratinizing (KSCC), non-keratinizing (NKSCC), and hybrid SCC (HSCC). EBV status was compared among various histological tumor entities and clinicopathological characteristics of OSCC using immunohistochemistry. Chi-square test was used to determine the association of each histological subtype with EBV status with P-value <0.05 considered as statistically significant. RESULTS: Conventional tumor was the most frequent squamous cell carcinoma (n=126; 84%). A significant statistical link of EBV infection was observed with rare histological tumors exhibiting acantholysis (P=0.01), as well as tumors involving buccal mucosa (P=0.03), and habitual smokers (P=0.001). CONCLUSIONS: In this study, acantholytic tumor, a rare histological subtype of OSCC, tended to be EBV related. Moreover, OSCC cases bearing EBV infection were more likely smokers favoring buccal mucosa as primary anatomical site for oral cancer.
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Escherichia coli is used intensively for recombinant protein production, but one key challenge with recombinant E. coli is the tendency of recombinant proteins to misfold and aggregate into insoluble inclusion bodies (IBs). IBs contain high concentrations of inactive recombinant protein that require recovery steps to salvage a functional recombinant protein. Currently, no universally effective method exists to prevent IB formation in recombinant E. coli. In this study, DNA microarrays were used to compare the E. coli gene expression response dynamics to soluble and insoluble recombinant protein production. As expected and previously reported, the classical heat-shock genes had increased expression due to IB formation, including protein folding chaperones and proteases. Gene expression levels for protein synthesis-related and energy-synthesis pathways were also increased. Many transmembrane transporter and corresponding catabolic pathways genes had decreased expression for substrates not present in the culture medium. Additionally, putative genes represented over one-third of the genes identified to have significant expression changes due to IB formation, indicating many important cellular responses to IB formation still need to be characterized. Interestingly, cells grown in 3% ethanol had significantly reduced gene expression responses due to IB formation. Taken together, these results indicate that IB formation is complex, stimulates the heat-shock response, increases protein and energy synthesis needs, and streamlines transport and catabolic processes, while ethanol diminished all of these responses.
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Proteínas de Escherichia coli/classificação , Proteínas de Escherichia coli/genética , Escherichia coli/genética , Corpos de Inclusão/metabolismo , Proteínas Recombinantes/metabolismo , Biotecnologia , Escherichia coli/efeitos dos fármacos , Escherichia coli/metabolismo , Proteínas de Escherichia coli/análise , Proteínas de Escherichia coli/metabolismo , Etanol/farmacologia , Perfilação da Expressão Gênica , Proteínas de Choque Térmico/genética , Proteínas de Choque Térmico/metabolismo , Redes e Vias Metabólicas/genética , Análise de Sequência com Séries de Oligonucleotídeos , SolubilidadeRESUMO
High quality, intact messenger RNA (mRNA) is required for DNA microarray and reverse transcriptase polymerase chain reaction analysis and is generally obtained from total RNA isolations. The most widely recognized measure of RNA integrity is the RNA Integrity Number (RIN) obtained from the Agilent Bioanalyzer, as it provides sizing, quantification, and quality control measures. This work describes comparisons of the RIN values obtained for recombinant E. coli. Uninduced recombinant E. coli cultures were examined, as well as induced cultures that produced either a soluble or insoluble recombinant protein. The uninduced cultures and the induced cultures producing soluble protein had higher RIN values than the induced cultures producing insoluble protein. These lower RIN values for E. coli producing the insoluble protein indicate that cellular degradation of the ribosomal RNA species is the likely cause of the lower RIN values. As the use of DNA microarrays and other gene expression tools increase in usage in the industrial recombinant protein production community, these results suggest the need for further studies to determine acceptable RIN ranges for gene expression analysis and effects of various culture conditions on RIN values for recombinant E. coli.
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OBJECTIVE: Xenotropic murine leukemia virus related virus (XMRV), is the first gammaretrovirus identified a decade ago, in human tissue bearing adenocarcinoma of prostate, followed by several researches documenting little or no prevalence of XMRV in prostate cancer samples. However, the status of XMRV within subtype of prostate adenocarcinoma has not been investigated yet. In this study, we investigated the relationship between XMRV and broad spectrum morphological entities of prostate adenocarcinoma, including acinar, ductal and other rare subtypes. MATERIAL AND METHODS: The prevalence of XMRV DNA in different histological subtypes of prostate adenocarcinoma was examined after characterizing the tumors into groups, using formalin-fixed, paraffin-embedded tissue samples from newly diagnosed prostate adenocarcinomas and archival prostate cancer tissue from our XMRV case control analysis. Broad-spectrum XMRV DNA amplification was performed by end-point polymerase chain reaction, using commercially available primer set. RESULTS: The study included 100 patients with prostate cancer. XMRV DNA was detected in 4 of 8 (50%) ductal adenocarcinomas, exhibiting papillary and cribriform histological features. XMRV DNA was not detected in any other variant of adenocarcinoma including acinar (0/91) and mucinous carcinomas (0/1). Majority of XMRV positive cases were biologically aggressive and present cancer at an early age upon diagnosis. CONCLUSION: Ductal adenocarcinomas demonstrate a significant association of XMRV DNA while other histological variants of prostate adenocarcinoma seem unrelated to XMRV infection.
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OBJECTIVES: We sought to characterize the ductal and acinar subtype of prostate adenocarcinoma using hematoxylin and eosin (H&E) staining and an immunohistochemical antibody cocktail. We also investigated the clinical features, prostate-specific antigen (PSA) levels, and biological aggressiveness of these tumors. METHODS: We utilized tumor bearing prostate biopsies, obtained between 2010 and 2014 from Dow Diagnostic Research and Reference Laboratory, to identify cases of prostatic ductal and acinar adenocarcinoma using routine H&E and immunohistochemical staining. The immunohistochemical antibody cocktail 34ßE12/p63/AMACR was used for staining. The association of clinicopathological variables including patient's age at diagnosis, Gleason score, and PSA levels before surgery was retrospectively analyzed. RESULTS: A total of 10 ductal and 140 non-ductal cases were identified. Ductal cases were predominantly high grade with advanced histopathological features (90%; p=0.030). Marked elevation in PSA level was also reported in most cases. No other significant statistical difference was observed. CONCLUSIONS: Pathological and immunohistochemical examination could be used to characterize ductal and acinar adenocarcinoma of the prostate. Ductal adenocarcinoma of the prostate is a rare subtype of prostate carcinoma and is be more likely to present with advanced grade cancer suggesting that timely detection of the disease is vital.
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OBJECTIVE: To determine the association of Xenotropic murine leukemia virus related virus (XMRV) infection with prostate cancer and compare it with benign prostate hyperplasia. STUDY DESIGN: Case control study. PLACE AND DURATION OF STUDY: Department of Histopathology and Molecular Pathology, Dow University of Health Sciences, Karachi, from January 2009 to December 2012. METHODOLOGY: XMRV was screened in 50 prostate cancer and 50 benign prostatic hyperplasia biopsies using conventional end-point PCR. Other studied variables were family history of prostate cancer, patients age and Gleason score. RESULTS: XMRV was detected in 4 (8%) of the 50 prostate cancer biopsy specimens compared to none in biopsies with benign prostatic hyperplasia. However, there was no significant statistical association of XMRV infection with the other variables. CONCLUSION: A low frequency of XMRV infection was found in this case-control study. Men, who harbor XMRV infection, may be at increased risk of prostate cancer but this needs to be investigated further at a larger scale.
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Adenocarcinoma/virologia , Próstata/virologia , Hiperplasia Prostática/virologia , Neoplasias da Próstata/virologia , Infecções Tumorais por Vírus/epidemiologia , Vírus Relacionado ao Vírus Xenotrópico da Leucemia Murina/isolamento & purificação , Adenocarcinoma/epidemiologia , Adenocarcinoma/patologia , Idoso , Idoso de 80 Anos ou mais , Biópsia , Estudos de Casos e Controles , Humanos , Masculino , Pessoa de Meia-Idade , Gradação de Tumores , Paquistão/epidemiologia , Reação em Cadeia da Polimerase , Próstata/patologia , Hiperplasia Prostática/epidemiologia , Hiperplasia Prostática/patologia , Neoplasias da Próstata/epidemiologia , Neoplasias da Próstata/patologia , Provírus/genética , RNA Viral/genética , RNA Viral/isolamento & purificação , Infecções Tumorais por Vírus/virologia , Vírus Relacionado ao Vírus Xenotrópico da Leucemia Murina/genéticaRESUMO
During locomotion over land, vertebrates' limb bones are exposed to loads. Like most biological structures, limb bones have a capacity to withstand greater loads than they usually experience, termed a safety factor (SF). How diverse are limb-bone SFs, and what factors correlate with such variation? We have examined these questions from two perspectives. First, we evaluated locomotor SF for the femur in diverse lineages, including salamanders, frogs, turtles, lizards, crocodilians, and marsupials (opossums). Comparisons with values for hind-limb elements in running birds and eutherian mammals indicate phylogenetic diversity in limb-bone SF. A high SF (â¼7) is primitive for tetrapods, but low magnitudes of load and elevated strength of bones contribute to different degrees across lineages; moreover, birds and eutherians appear to have evolved lower SFs independently. Second, we tested the hypothesis that SFs would be similar across limb bones within a taxon by comparing data from the humerus and femur of alligators. Both in bending and in torsion, we found a higher SF for the humerus than for the femur. Such a "mixed chain" of different SFs across elements has been predicted if bones have differing variabilities in load, different costs to maintain, or high SF values in general. Although variability in load is similar for the humerus and femur, a high SF may be less costly for the humerus because it is smaller than the femur. The high SFs of alligators also might facilitate differences in SF among their limb bones. Beyond these specific findings, however, a more general implication of our results is that evaluations of the diversity of limb-bone SFs can provide important perspective to direct future research. In particular, more complete understanding of variation in SF could provide insight into factors that promoted the evolutionary radiation of terrestrial locomotor function in vertebrates.