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BACKGROUND: The aim of this study was to investigate the impact of hMLH1 polymorphisms on treatment outcomes in patients with oral squamous cell carcinoma (OSCC). METHODS: Genotypings were performed by direct DNA sequencing in peripheral blood leukocytes from 185 male OSCC patients. Patients received primary surgery with or without adjuvant radiotherapy. Two hMLH1 tag single nucleotide polymorphisms (SNPs)-rs1800734 (-93G>A in the promoter) and rs1540354 (in the third intron)-were chosen from the HapMap project. Overall survival (OS) and disease-free survival (DFS) were compared between different genotypes. RESULTS: The hMLH1 rs1800734 and rs1540354 polymorphisms were in weak linkage disequilibrium (r (2) = 0.456). OSCC patients with the rs1800734 AA genotype had a significantly poor prognosis in both OS and DFS. This SNP can also predict the outcomes of OSCC patients with postoperative adjuvant radiotherapy, especially in advanced stage; however, no significant differences in patient outcomes were found for the hMLH1 rs1540354 genotypes. CONCLUSIONS: Our results demonstrate that the hMLH1 -93G>A SNP is found to be associated with patient outcomes in OSCC. This SNP can also predict their treatment outcome of radiotherapy.
Assuntos
Proteínas Adaptadoras de Transdução de Sinal/genética , Biomarcadores Tumorais/genética , Carcinoma de Células Escamosas/genética , Neoplasias Bucais/genética , Proteínas Nucleares/genética , Polimorfismo de Nucleotídeo Único/genética , Regiões Promotoras Genéticas/genética , Carcinoma de Células Escamosas/mortalidade , Carcinoma de Células Escamosas/patologia , Seguimentos , Predisposição Genética para Doença , Genótipo , Humanos , Masculino , Pessoa de Meia-Idade , Neoplasias Bucais/mortalidade , Neoplasias Bucais/patologia , Proteína 1 Homóloga a MutL , Gradação de Tumores , Estadiamento de Neoplasias , Prognóstico , Taxa de SobrevidaRESUMO
AIMS: To investigate the relationship of matriptase expression in oral squamous cell carcinoma (OSCC) to clinicopathological characteristics, patient survival and cell-invasive properties. METHODS AND RESULTS: Matriptase expression in OSCC was evaluated by immunohistochemical staining, and its relationship to clinicopathological features and outcomes was assessed statistically. The shRNA-mediated stable knockdown of matriptase in OSCC cells was used to analyse cell proliferation, migration and invasion in vitro. Matriptase immunostaining score was correlated with histopathological grade, clinical stage, positive lymph node and distant metastasis, and higher matriptase immunostaining score was associated significantly with poor prognosis. Elevated matriptase expression in oral cancer cell lines was a significant promoter of oral cancer cell migration and invasion. CONCLUSIONS: Matriptase expression correlates with tumour progression and invasive capability in OSCC and may be an adverse prognostic marker for this cancer.
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Biomarcadores Tumorais/análise , Carcinoma de Células Escamosas/patologia , Neoplasias Bucais/patologia , Serina Endopeptidases/biossíntese , Adulto , Idoso , Western Blotting , Carcinoma de Células Escamosas/enzimologia , Carcinoma de Células Escamosas/mortalidade , Movimento Celular/fisiologia , Proliferação de Células/fisiologia , Progressão da Doença , Feminino , Técnicas de Silenciamento de Genes , Humanos , Imuno-Histoquímica , Estimativa de Kaplan-Meier , Masculino , Pessoa de Meia-Idade , Neoplasias Bucais/enzimologia , Neoplasias Bucais/mortalidade , Prognóstico , Modelos de Riscos Proporcionais , RNA Interferente PequenoRESUMO
Background/purpose: The serpin peptidase inhibitor, clade E, member 2 (SERPINE2), is upregulated in breast cancer, prostate cancer, and urothelial carcinoma; however, limited information exists regarding its expression in oral cancer. Therefore, this study aimed to analyze the association between SERPINE2 expression and oral squamous cell carcinoma (OSCC) outcomes. Materials and methods: SERPINE2 mRNA and protein expression in patients with head and neck squamous cell carcinoma and OSCC were investigated using online databases and tissue-array analysis. Its relationship with clinicopathological characteristics, OSCC prognosis and its biological function in OSCC cells were explored. Results: Analysis using online databases revealed higher SERPINE2 expression in tumor tissues and its role as a prognostic factor. High SERPINE2 protein levels were significantly correlated with adverse pathological parameters, including advanced clinical stage and tumor status (P < 0.001), lymph nodes (P = 0.014), and distant metastases (P = 0.013). High SERPINE2 expression was associated with worse overall survival (P < 0.001) and was identified as an independent prognostic factor for OSCC. In vitro studies revealed that SERPINE2 knockdown significantly reduced cell proliferation, migration, and invasion in OSCC cell lines. Conclusion: This study suggests that SERPINE2 may serve as a prognostic biomarker and potential therapeutic target for oral cancer.
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Recent studies have reported that SERPINE2 contributes to the development of various cancers. However, its association with urothelial carcinoma (UC) remains unclear. In this study, data on urinary bladder UC (UBUC) cases from The Cancer Genome Atlas (TCGA) database were used to investigate the prognostic value of SERPINE2 mRNA expression. Then, SERPINE2 expression was analyzed with tissue microarrays constructed from 117 upper tract UC (UTUC) and 84 UBUC tissue specimens using immunohistochemical staining. Results were compared to clinicopathologic data by multivariate analysis. In the TCGA database, high SERPINE2 mRNA expression indicated a poor prognosis in patients with UBUC. Furthermore, Mann-Whitney U test showed that high SERPINE2 immunoexpression was significantly associated with adverse pathologic parameters including invasion, high grade, coexistence of UC in situ, and advanced pT stage (all p < 0.05, except for a marginal association with high-grade UBUC, p = 0.066). Kaplan-Meier analysis revealed that high SERPINE2 expression was associated with worse overall survival (OS; UTUC, p = 0.003; UBUC, p = 0.014) and disease-free survival (UTUC, p = 0.031; UBUC, p = 0.033). Moreover, multivariate analysis identified high SERPINE2 expression as an independent prognostic factor for OS (UTUC, p = 0.002; UBUC, p = 0.024). Taken together, our findings demonstrated that increased SERPINE2 expression is associated with adverse pathologic features and may serve as a prognostic biomarker for UC.
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BACKGROUND: O (6) -methylguanine-DNA methyltransferase (MGMT) is a specific DNA direct reversal repair protein which ameliorates mutagenic, carcinogenic and cytotoxic adducts from O(6) -methylguanine in DNA. The aim of this study was to investigate the potential association between six polymorphisms in the coding region of the MGMT gene and oral cancer risk in Taiwanese population. METHODS: In this hospital-based case-control study, the MGMT genotypes were determined by using DNA sequencing in 176 patients with oral squamous cell carcinoma (OSCC), 77 patients with oral premalignant lesions and 110 normal individuals. RESULTS: Although L53L and L84F polymorphisms were in complete linkage disequilibrium, no statistically significant association was found between the MGMT genotypes and haplotypes and oral cancer risk. We could also not detect any variations in the codon 65, 143, 160 and 178 within the coding region of the MGMT gene in both patients and controls. After stratification of OSCC patients by their mean age (54 years old), however, a higher overall survival rate with CT genotype than CC genotypes of the L53L polymorphisms was found in older patients (P=0.031). A borderline significance (P=0.074) between these genotypes and recurrence-free survivals has also been noted. The same results were also observed in L84F polymorphism. CONCLUSIONS: The present study did not find statistically significant association between six common MGMT polymorphisms and oral cancer risk, however, both MGMT L53L and L84F polymorphisms in old patients with CT genotype have higher overall survival rates than patients with CC genotype. Because of limited power of the present study, further study may be warranted in ethnically different populations to explore outcomes of these polymorphisms in the oral cancer risk.
Assuntos
Códon/genética , Predisposição Genética para Doença/genética , Neoplasias Bucais/genética , O(6)-Metilguanina-DNA Metiltransferase/genética , Polimorfismo Genético/genética , Fatores Etários , Carcinoma de Células Escamosas/genética , Estudos de Casos e Controles , Citosina , Intervalo Livre de Doença , Feminino , Frequência do Gene/genética , Variação Genética/genética , Genótipo , Haplótipos/genética , Humanos , Leucina/genética , Desequilíbrio de Ligação/genética , Masculino , Pessoa de Meia-Idade , Fenilalanina/genética , Polimorfismo de Nucleotídeo Único/genética , Lesões Pré-Cancerosas/genética , Estudos Retrospectivos , Fatores de Risco , Análise de Sequência de DNA , Taxa de Sobrevida , Taiwan , TiminaRESUMO
A novel one-step, closed-tube, loop-mediated isothermal amplification (LAMP) assay for detecting Entamoeba histolytica, one of the leading causes of morbidity in developing countries, was developed. The sensitivity of the LAMP assay is 1 parasite per reaction. A total of 130 clinical samples were analyzed, and the results compared with those of conventional nested PCR to validate the practicability of this assay. No DNA was amplified from other diarrheal pathogens, such as other Entamoeba species, bacteria, and viruses. These results indicate that LAMP is a rapid, simple, and valuable diagnostic tool for epidemiological studies of amebiasis.
Assuntos
Entamoeba histolytica/isolamento & purificação , Entamebíase/diagnóstico , Técnicas de Amplificação de Ácido Nucleico/métodos , Animais , Entamoeba histolytica/genética , Entamebíase/parasitologia , Humanos , Sensibilidade e EspecificidadeRESUMO
The aim of this study was to investigate the relationship of matriptase-2 expression with the clinicopathologic characteristics, the histologic grade, and patient survival in oral squamous-cell carcinoma (OSCC). Immunohistochemical analysis of matriptase-2 expression was performed in 102 surgical specimens from patients with OSCC. The immunohistochemical results were further verified by quantitative real-time reverse transcription-polymerase chain reaction. The immunostaining intensity was scored on a scale ranging from 0 (absence of staining) to 3 (intense staining). The distribution score was determined by the percentage of stained cells on a scale ranging from 0 (<5%), 1 (5% to 25%), 2 (25% to 50%), 3 (50% to 75%), to 4 (75% to 100%). The immunoscore of matriptase-2 expression was the product of the above 2 scores and ranged from 0 to 12 for analysis. Faint matriptase-2 immunostaining was observed in the non-neoplastic oral mucosal epithelia. The matriptase-2 immunoscore was significantly higher in well-differentiated OSCCs than in poorly differentiated tumors (P=0.001). Moreover, a reduced matriptase-2 immunoscore was inversely correlated with the tumor size (P=0.017), a positive nodal stage (P=0.008), distant metastasis (P=0.032), and a late clinical stage (P=0.001). A lower immunoscore of matriptase-2 expression revealed a significant association with poor survival (P=0.003). Our results demonstrate that the inverse expression of matriptase-2 correlates with tumor progression and an advanced TNM stage, and has a poor prognosis in patients with OSCC. These findings suggest that the expression of matriptase-2 may be both a prognostic marker and a potential therapeutic target for this cancer.
Assuntos
Biomarcadores Tumorais/biossíntese , Carcinoma de Células Escamosas , Regulação para Baixo , Regulação Enzimológica da Expressão Gênica , Regulação Neoplásica da Expressão Gênica , Proteínas de Membrana/biossíntese , Neoplasias Bucais , Proteínas de Neoplasias/biossíntese , Serina Endopeptidases/biossíntese , Adulto , Idoso , Carcinoma de Células Escamosas/enzimologia , Carcinoma de Células Escamosas/mortalidade , Carcinoma de Células Escamosas/patologia , Intervalo Livre de Doença , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Neoplasias Bucais/enzimologia , Neoplasias Bucais/mortalidade , Neoplasias Bucais/patologia , Taxa de SobrevidaRESUMO
BACKGROUND: Apurinic/apyrimidinic endonuclease 1/redox factor-1 (APE1/Ref-1) is a multifunctional protein involved in DNA repair and redox signaling. The purpose of this study was to investigate the relationship between APE1/Ref-1 expression and clinicopathological features, survival, and treatment response in patients with oral squamous cell carcinoma (OSCC) and cell lines. METHODS: APE1/Ref-1 expression in OSCC was evaluated by immunohistochemistry, and its relationship to patient outcomes and treatment response was assessed statistically. The effects of stable short hairpin (sh)RNA-mediated knockdown of APE1/Ref-1 on cell survival, migration, and chemoradiation sensitivity were determined in OSCC cell lines. RESULTS: APE1/Ref-1 immunostaining was correlated with positive lymph node status, and higher APE1/Ref-1 expression was significantly associated with poor prognosis and reduced treatment response. Consistent with the clinical studies, APE1/Ref-1 expression in OSCC cell lines was implicated in the regulation of migration and cisplatin-induced apoptosis. CONCLUSION: Elevated APE1/Ref-1 expression may be used to predict poor survival and may confer chemoresistance in OSCC.
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Carcinoma de Células Escamosas/mortalidade , DNA Liase (Sítios Apurínicos ou Apirimidínicos)/metabolismo , Neoplasias Bucais/mortalidade , Apoptose , Western Blotting , Carcinoma de Células Escamosas/metabolismo , Linhagem Celular Tumoral , Ensaios de Migração Celular , Sobrevivência Celular , Feminino , Técnicas de Silenciamento de Genes , Humanos , Imuno-Histoquímica , Masculino , Pessoa de Meia-Idade , Neoplasias Bucais/metabolismo , RNA Interferente Pequeno/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Taxa de Sobrevida , Resultado do TratamentoRESUMO
Class switch recombination (CSR) is a programmed gene rearrangement in which a B cell which is producing IgM and IgD antibody develops into an IgG-, IgA- or IgE-expressing cell. This is achieved by recombination between switch regions located 5' of each of the immunoglobulin heavy chain constant regions, except Cdelta. The mechanism of CSR has not been resolved but it is thought to involve a double-strand break followed by end joining. It has previously been suggested that the nucleotide excision repair protein ERCC1 may be involved in CSR due to its known roles in removal of 3' single-stranded tails in various types of recombination. In this study, we examined class switching in cultured splenocytes from ERCC1-deficient mice and found no evidence of any deficiency.
Assuntos
Reparo do DNA , Proteínas de Ligação a DNA , Endonucleases , Switching de Imunoglobulina , Proteínas/fisiologia , Recombinação Genética , Animais , Linfócitos B/metabolismo , Sequência de Bases , Citometria de Fluxo , Genótipo , Camundongos , Camundongos Transgênicos , Modelos Genéticos , Dados de Sequência Molecular , Reação em Cadeia da Polimerase , Proteínas/metabolismo , Baço/citologia , Fatores de Tempo , TransgenesRESUMO
Gastric adenocarcinoma is a lethal disease with high incidence in Chinese people. The purpose of this study was to evaluate the expression of urocortin (UCN) in normal gastric mucosa and gastric adenocarcinomas. Immunohistochemical analysis of UCN was performed in 112 surgical specimens (21 normal gastric mucosa specimens and 91 gastric adenocarcinoma specimens varying in histologic grade and pathologic stage). Immunostain intensity was scored on a scale ranging from 0 (absence of staining) to 3 (strong staining). The percentage of UCN stained cells was scored on a scale ranging from 0 (<5%) to 4 (75% to 100%). The UCN immunoscore (ranging from 0 to 12) was the product of the above 2 scores. The UCN immunoscore was high in all 9 normal gastric mucosa specimens, significantly lower in poorly differentiated gastric adenocarcinoma than in well and moderately differentiated tumors (P=0.018), and significantly lower in more advanced pathologic stages of gastric adenocarcinomas than in the early stages of these tumors. Moreover, UCN expression was higher in gastric adenocarcinomas with neuroendocrine differentiation than in mucinous adenocarcinomas and signet-ring cell carcinomas. In conclusion, UCN is expressed in most non-neoplastic gastric glandular epithelia. However, UCN expression inversely correlates with higher tumor grade and advanced TNM stage in gastric adenocarcinomas.
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Adenocarcinoma Mucinoso/genética , Adenocarcinoma/genética , Carcinoma de Células em Anel de Sinete/genética , Neoplasias Gástricas/genética , Urocortinas/genética , Adenocarcinoma/diagnóstico , Adenocarcinoma/patologia , Adenocarcinoma/cirurgia , Adenocarcinoma Mucinoso/diagnóstico , Adenocarcinoma Mucinoso/patologia , Adenocarcinoma Mucinoso/cirurgia , Carcinoma de Células em Anel de Sinete/diagnóstico , Carcinoma de Células em Anel de Sinete/patologia , Carcinoma de Células em Anel de Sinete/cirurgia , Feminino , Mucosa Gástrica/metabolismo , Expressão Gênica , Humanos , Imuno-Histoquímica , Masculino , Gradação de Tumores , Estadiamento de Neoplasias , Estômago/patologia , Neoplasias Gástricas/diagnóstico , Neoplasias Gástricas/patologia , Neoplasias Gástricas/cirurgia , Análise Serial de TecidosRESUMO
Urocortin (UCN) is a 40-amino acid neuropeptide that regulates angiogenesis and inhibits cell proliferation. Our aim was to examine the relationship of UCN expression to the clinicopathological parameters of pancreatic ductal adenocarcinoma (PDAC) and histological grade of pancreatic intraepithelial neoplasia (PanIN). Tissue microarray was used to analyze UCN protein expression in 89 surgical specimens including 21 PanIN, 3 PDAC arising from PanIN, and 65 PDAC without PanIN. UCN immunoscores ranging from 0 to 12 were obtained by multiplying intensity (scored on a 3-point scale) by the percentage of stained cells (scored on a 4-point scale). Strong expression of UCN was detected in 5 specimens of non-neoplastic pancreatic ductal epithelia. UCN immunoscore was significantly higher in PanIN-1 than in PanIN-2 and PanIN-3 (p = 0.038) and significantly higher in well-differentiated PDAC or early American Joint Committee on Cancer (AJCC) stage PDAC than in poorly differentiated or advanced stage PDAC (p = 0.025, p = 0.018). Higher expression of UCN correlates with PDAC tumor grade and AJCC pathologic stage as well as PanIN grade. Immunohistochemical assessment of UCN may help clinicians predict tumor recurrence rate and help pathologists make a proper diagnosis.
Assuntos
Carcinoma in Situ/genética , Carcinoma Ductal Pancreático/genética , Regulação Neoplásica da Expressão Gênica , Neoplasias Pancreáticas/genética , Urocortinas/metabolismo , Adulto , Idoso , Biomarcadores Tumorais/genética , Biomarcadores Tumorais/metabolismo , Carcinoma in Situ/diagnóstico , Carcinoma in Situ/patologia , Carcinoma Ductal Pancreático/diagnóstico , Carcinoma Ductal Pancreático/patologia , Proliferação de Células , Feminino , Humanos , Imuno-Histoquímica , Masculino , Pessoa de Meia-Idade , Neoplasias Pancreáticas/diagnóstico , Neoplasias Pancreáticas/patologia , Análise Serial de Tecidos , Urocortinas/genéticaRESUMO
The objective of this study was to analyze recent infections and the molecular epidemiology of human immunodeficiency virus type 1 (HIV-1) among different risk groups since the outbreak of circulating recombinant form CRF07_BC among intravenous drug users (IDUs) in 2004 in Taiwan. Phylogenetic analysis was performed using the env and pol fragment sequences amplified from these specimens. The BED IgG capture incidence EIA (BED-CEIA assay) was used to determine recent infections. Among the 683 HIV-1-positive individuals enrolled between 2007 and 2009, 394 (57.7%) were subtype B, 260 (38.1%) were CRF07_BC, 26 (3.8%) were CRF01_AE, two (0.3%) were CRF08_BC, and one (0.1%) was CRF06_cpx. While the percentage of CRF07_BC decreased (58.5-17.9%, p < 0.001) from 2007 to 2009, the percentage of subtype B increased (37.6% to 74.9%, p < 0.001). A concordant decrease in the proportion of recent infections to new infections among IDUs (63.6% to 9.8%, p < 0.001), accompanied with an increase of the proportion of recent infections in MSM (men having sex with men) (22.4-67.1%, p = 0.77) and heterosexual groups (13.1- 23.2%, p = 0.852), was observed. The decrease in CRF07_BC infections and the reduction in the proportion of recent infections among IDUs reflected the success of harm reduction strategies initiated by the government in 2005.
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Surtos de Doenças , Infecções por HIV/epidemiologia , Infecções por HIV/virologia , HIV-1/classificação , HIV-1/genética , Adulto , Análise por Conglomerados , Estudos Transversais , Feminino , Genótipo , HIV-1/isolamento & purificação , Humanos , Masculino , Pessoa de Meia-Idade , Epidemiologia Molecular , Dados de Sequência Molecular , Filogenia , Assunção de Riscos , Análise de Sequência de DNA , Taiwan/epidemiologia , Produtos do Gene env do Vírus da Imunodeficiência Humana/genética , Produtos do Gene pol do Vírus da Imunodeficiência Humana/genéticaRESUMO
This article describes a case of human immunodeficiency virus type 1 (HIV-1) infection transmission caused by a bloody knife fight in a robbery. The victim was a 69-year-old man who was not infected with HIV-1, and his wife was HIV-antibody negative. A robber, a 42-year-old man, was HIV antibody-positive since December 2005 and had not taken antiretroviral therapy. The BED IgG Capture incidence EIA (BED-CEIA assay) data showed that the specimens from the victim were compatible with a recent seroconversion. Phylogenetic analysis of fragments of pol, encompassing protease and a portion of reverse transcriptase, and of env genes isolated from the victim, the robber, and a local population samples of HIV-1 positive individuals showed that the victim's HIV-1 sequences were most closely related to and nested within a lineage comprised of the robber's HIV-1 sequences. We provide HIV-1 seroconversion data and phylogenetic analysis as evidence that the HIV-1 transmission likely occurred from contact during the robbery.
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Infecções por HIV/transmissão , Ferimentos Perfurantes , Adulto , Idoso , Sequência de Bases , Clonagem Molecular , Primers do DNA , Infecções por HIV/virologia , HIV-1/classificação , HIV-1/isolamento & purificação , Humanos , Técnicas Imunoenzimáticas , Masculino , FilogeniaRESUMO
OBJECTIVE: O(6)-methylguanine-DNA methyltransferase (MGMT) ameliorates mutagenic, carcinogenic, and cytotoxic adducts from O(6)-methylguanine in DNA through a direct reversal mechanism. Decreased expression of MGMT has been reported in a variety of human malignant tumors. The purpose of this study was to clarify the correlation of MGMT expression levels in oral squamous cell carcinoma (OSCC) with promoter hypermethylation and with betel quid chewing and cigarette smoking. STUDY DESIGN: MGMT protein expression in 63 cases of oral squamous cell carcinoma by immunohistochemistry was investigated. Methylation status of the MGMT was analyzed by methylation-specific PCR. Correlation with clinicopathologic parameters was then tested by statistical analysis. RESULTS: MGMT immunohistochemistry revealed nuclear staining in normal epithelium, whereas 47 (75%) of 63 OSCC tumors were devoid of MGMT expression and this was related to tumor cell differentiation. Furthermore, the association between loss of MGMT expression and promoter hypermethylation was significant. Lacking protein expression for MGMT in OSCC was also associated with the use of betel quid. CONCLUSIONS: The results suggest that the absence of MGMT expression, which would seem to be associated with promoter hypermethylation, is related to betel quid chewing and, thus, in turn, might be a significant event in oral carcinogenesis.
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Areca/efeitos adversos , Carcinoma de Células Escamosas/enzimologia , Neoplasias Bucais/enzimologia , O(6)-Metilguanina-DNA Metiltransferase/metabolismo , Regiões Promotoras Genéticas/efeitos dos fármacos , Adulto , Feminino , Humanos , Imuno-Histoquímica , Masculino , Metilação , Pessoa de Meia-Idade , O(6)-Metilguanina-DNA Metiltransferase/efeitos dos fármacos , Fumar/efeitos adversos , Estatísticas não ParamétricasRESUMO
A single-tube multiprobe real-time PCR assay for simultaneous detection of Entamoeba histolytica and Entamoeba dispar was developed. One primer pair with 2 species-specific probes was designed based on new SSU RNA regions of the ribosomal DNA-containing episome. The sensitivity is 1 parasite per milliliter of feces and thus superior to the conventional nested PCR and comparable to other published real-time PCR protocols. The applicability for clinical diagnosis was validated with 218 stool specimens from patients. A total of 51 E. histolytica and 39 E. dispar positive samples was detected by the multiprobe real-time PCR compared to 39 and 22 by routine nested PCR diagnosis. The detection rate of Entamoeba species for the multiprobe real-time PCR assays was significantly higher than the nested PCR (40.8% vs. 28.0%, P < 0.01). The test did not show cross reactivity with DNA from Entamoeba moshkovskii, Giardia lamblia , Cryptosporidium sp., Escherichia coli , or other nonpathogenic enteric parasites. The multiprobe real-time PCR assay is simple and rapid and has high specificity and sensitivity. The assay could streamline the laboratory diagnosis procedure and facilitate epidemiological investigation.
Assuntos
DNA de Protozoário/análise , Entamoeba/isolamento & purificação , Entamebíase/diagnóstico , Reação em Cadeia da Polimerase/métodos , Primers do DNA/normas , Sondas de DNA/normas , Eletroforese em Gel de Ágar , Entamoeba/classificação , Entamoeba/genética , Entamoeba histolytica/classificação , Entamoeba histolytica/genética , Entamoeba histolytica/isolamento & purificação , Entamebíase/parasitologia , Fezes/parasitologia , Humanos , Modelos Lineares , Reação em Cadeia da Polimerase/normas , Sensibilidade e Especificidade , Especificidade da EspécieRESUMO
The relationship between histone methylation and apoptosis, programmed cell death, is beginning to be explored. The objective of this study was to investigate the effects of staurosporine, a PKC inhibitor on the methylation of histone H3 in osteosarcoma cells. Following stimulation by staurosporine in vitro of G292 cells, a human osteosarcoma cell line with fibroblast-like phenotype, methylation of histone H3 was evaluated by western blotting and immunocytochemistry. G292 cells revealed the expression of cleaved PARP after incubation with staurosporine for 3 hours. Monomethyl lysine (K) 27 was induced by staurosporine at a concentration of 1, but no monomethyl K4 or K9 in histone H3 was seen. Dimethyl and trimethyl histone H3 K27 were also identified. There was no expression of dimethyl or trimethyl histone H3 K4 and K9. Expression of monomethyl histone H3 K27 was dose-dependent. The morphologic changes of apoptosis induced by staurosporine were observed under microscopy. Immunocytochemistry of monomethyl histone H3 K27 showed a weak signal in controls, a strong signal in staurosporine-treated tumor cells and a denser signal in the apoptotic cells. Our studies demonstrated that monomethyl histone H3 lysine 27 is expressed in staurosporine-induced apoptotic osteosarcoma cells. The findings may provide novel bridge information between the epigenetic episodes and apoptotic process.
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Apoptose/efeitos dos fármacos , Inibidores Enzimáticos/farmacologia , Histonas/metabolismo , Osteossarcoma/metabolismo , Estaurosporina/farmacologia , Linhagem Celular Tumoral , Relação Dose-Resposta a Droga , Epigênese Genética/efeitos dos fármacos , Fluoresceína/metabolismo , Imunofluorescência , Corantes Fluorescentes/metabolismo , Expressão Gênica , Humanos , Imuno-Histoquímica , Metilação/efeitos dos fármacos , Osteossarcoma/genética , Osteossarcoma/patologia , Poli(ADP-Ribose) Polimerases/metabolismo , Proteína Quinase C/antagonistas & inibidoresRESUMO
Ercc1 is essential for nucleotide excision repair (NER) but, unlike other NER proteins, Ercc1 and Xpf are also involved in recombination repair pathways. Ercc1 knockout mice have profound cell cycle abnormalities in the liver and die before weaning. Subsequently Xpa and Xpc knockouts have proved to be good models for the human NER deficiency disease, xeroderma pigmentosum, leading to speculation that the recombination, rather than the NER deficit is the key to the Ercc1 knockout phenotype. To investigate the importance of the recombination repair functions of Ercc1 we studied spermatogenesis and oogenesis in Ercc1-deficient mice. Male and female Ercc1-deficient mice were both infertile. Ercc1 was expressed at a high level in the testis and the highest levels of Ercc1 protein occurred in germ cells following meiotic crossing over. However, in Ercc1 null males some germ cell loss occurred prior to meiotic entry and there was no evidence that Ercc1 was essential for meiotic crossing over. An increased level of DNA strand breaks and oxidative DNA damage was found in Ercc1-deficient testis and increased apoptosis was noted in male germ cells. We conclude that the repair functions of Ercc1 are required in both male and female germ cells at all stages of their maturation. The role of endogenous oxidative DNA damage and the reason for the sensitivity of the germ cells to Ercc1 deficiency are discussed.