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Background: Coronavirus disease 2019 (COVID-19), caused by severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2), immediately became a pandemic. Therefore, nosocomial infection control is necessary to screen for patients with possible COVID-19. Objective: This study aimed to investigate commonly measured clinical variables to predict COVID-19. Methods: This cross-sectional study enrolled 1087 patients in the isolation ward of a university hospital. Conferences were organized to differentiate COVID-19 from non-COVID-19 cases, and multiple nucleic acid tests were mandatory when COVID-19 could not be excluded. Multivariate logistic regression models were employed to determine the clinical factors associated with COVID-19 at the time of hospitalization. Results: Overall, 352 (32.4%) patients were diagnosed with COVID-19. The majority of the non-COVID-19 cases were predominantly caused by bacterial infections. Multivariate analysis indicated that COVID-19 was significantly associated with age, sex, body mass index, lactate dehydrogenase, C-reactive protein, and malignancy. Conclusion: Some clinical factors are useful to predict patients with COVID-19 among those with symptoms similar to COVID-19. This study suggests that at least two real-time reverse-transcription polymerase chain reactions of SARS-CoV-2 are recommended to exclude COVID-19.
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ãThe issue of drug lag in Japan has been rapidly reduced in recent years, and newly approved drugs now become available on Japanese and international markets at the same time. In this context, the risk management plan (RMP) system was introduced in 2012. RMPs describe important safety concerns recognized by Japan's Pharmaceuticals and Medical Devices Agency (PMDA) and marketing authorization holders (MAHs), as well as safety measures that MAHs request healthcare professionals (HCPs) to follow. The publication of RMPs is expected to support the sharing of drug risk management among HCPs during the postmarketing phase. In addition, to encourage risk communication between HCPs and patients, the PMDA website provides drug guides for patients and other information to promote proper understanding of drugs by patients and their families and enable them to identify serious adverse drug reactions at an early stage. However, the results of surveys conducted by the PMDA in FY2014 and FY2015 revealed low levels of awareness of RMPs and drug guides for patients in hospitals and other healthcare institutions. The surveys also showed that information regarding the proper use of drugs from MAHs and the PMDA was not incorporated into practice at healthcare institutions, resulting in the repeated release of identical safety alerts. To facilitate the increased utilization of risk communication tools, the PMDA has been providing and disseminating these tools through its website. This study addresses those efforts and the associated challenges.
Assuntos
Comunicação , Serviços de Informação sobre Medicamentos , Equipamentos e Provisões , Preparações Farmacêuticas , Gestão de Riscos , Aprovação de Drogas , Efeitos Colaterais e Reações Adversas Relacionados a Medicamentos , Órgãos Governamentais , Pessoal de Saúde , Humanos , JapãoRESUMO
INTRODUCTION: International norms and ethical standards have suggested that compensation for research-related injury should be provided to injured research volunteers. However, statistical data of incidence of compensation claims and the rate of awarding them have been rarely reported. METHOD: Questionnaire surveys were sent to pharmaceutical companies and medical institutions, focusing on industry-initiated clinical trials aiming at new drug applications (NDAs) on patient volunteers in Japan. RESULTS: With the answers from pharmaceutical companies, the incidence of compensation was 0.8%, including 0.06% of monetary compensation. Of the cases of compensation claims, 99% were awarded. In turn, with the answers from medical institutions, the incidence of compensation was 0.6%, including 0.4% of serious but not death cases, and 0.04% of death cases. Furthermore, most claims for compensation were initiated by medical institutions, rather than by the patients. On the other hand, with the answers from clinical trial volunteers, 3% of respondents received compensations. These compensated cases were 25% of the injuries which cannot be ruled out from the scope of compensation. CONCLUSION: Our study results demonstrated that Japanese pharmaceutical companies have provided a high rate of compensation for clinical trial-related injuries despite the possibility of overestimation. In the era of global clinical development, our study indicates the importance of further surveys to find each country's compensation policy by determining how it is being implemented based on a survey of the actual status of compensation coming from statistical data.
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Compensação e Reparação , Indústria Farmacêutica/economia , Voluntários Saudáveis/legislação & jurisprudência , Revisão da Utilização de Seguros/economia , Ferimentos e Lesões/economia , Ensaios Clínicos como Assunto , Indústria Farmacêutica/ética , Indústria Farmacêutica/estatística & dados numéricos , Drogas em Investigação/efeitos adversos , Humanos , Revisão da Utilização de Seguros/ética , Revisão da Utilização de Seguros/estatística & dados numéricos , Japão , Inquéritos e Questionários , Ferimentos e Lesões/induzido quimicamenteRESUMO
OBJECTIVES: Pancreatic cancer is one of the most lethal malignancies; its poor prognosis is strongly associated with invasion and metastasis. Expression of S100A4 has been reported to correlate with poor prognosis in various cancers. We have investigated the role of S100A4 in pancreatic cancer tumorigenesis and its clinicopathologic significance. METHODS: Protein expression of S100A4 was examined by Western blot in pancreatic cancer cell lines and a human pancreatic ductal epithelium cell line, HPDE-6. Then the expressions of S100A4, TP53, and CD133 were examined immunohistochemically in resected specimens from 83 patients with pancreatic cancer to clarify their clinicopathologic significance. Survival analyses were performed using the Kaplan-Meier method and the Mantel-Cox method. RESULTS: Forty-eight (58%) of 83 patients with pancreatic cancer positively expressed S100A4, and 50 (60%) and 29 (36%) patients positively expressed TP53 and CD133, respectively. S100A4 expression was significantly correlated with perineural invasion (P = 0.029) and invasion pattern (P = 0.001). Neither TP53 nor CD133 expression showed significant correlations with any other parameters. CONCLUSIONS: Our present results suggest that S100A4 plays an important role in the invasiveness, particularly with perineural invasion and invasion pattern, of pancreatic cancer. Development of new strategies targeting S100A4 or its downstream effectors is warranted.
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Adenocarcinoma/metabolismo , Carcinoma Ductal Pancreático/metabolismo , Neoplasias Pancreáticas/metabolismo , Proteínas S100/biossíntese , Antígeno AC133 , Adenocarcinoma/patologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Antígenos CD/metabolismo , Western Blotting , Carcinoma Ductal Pancreático/patologia , Linhagem Celular Tumoral , Glicoproteínas/metabolismo , Humanos , Imuno-Histoquímica , Estimativa de Kaplan-Meier , Pessoa de Meia-Idade , Invasividade Neoplásica , Neoplasias Pancreáticas/patologia , Peptídeos/metabolismo , Prognóstico , Proteína A4 de Ligação a Cálcio da Família S100 , Análise Serial de Tecidos , Proteína Supressora de Tumor p53/metabolismoRESUMO
Anthocyanin O-methyltransferase (OMT) is one of the key enzymes for anthocyanin modification and flower pigmentation. We previously bred a novel red-purple-flowered fragrant cyclamen (KMrp) from the purple-flowered fragrant cyclamen 'Kaori-no-mai' (KM) by ion-beam irradiation. Since the major anthocyanins in KMrp and KM petals were delphinidin 3,5-diglucoside and malvidin 3,5-diglucoside, respectively, inactivation of a methylation step in the anthocyanin biosynthetic pathway was indicated in KMrp. We isolated and compared OMT genes expressed in KM and KMrp petals. RT-PCR analysis revealed that CkmOMT2 was expressed in the petals of KM but not in KMrp. Three additional CkmOMTs with identical sequences were expressed in petals of both KM and KMrp. Genomic PCR analysis revealed that CkmOMT2 was not amplified from the KMrp genome, indicating that ion-beam irradiation caused a loss of the entire CkmOMT2 region in KMrp. In vitro enzyme assay demonstrated that CkmOMT2 catalyzes the 3' or 3',5' O-methylation of the B-ring of anthocyanin substrates. These results suggest that CkmOMT2 is functional for anthocyanin methylation, and defective expression of CkmOMT2 is responsible for changes in anthocyanin composition and flower coloration in KMrp.
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Antocianinas/biossíntese , Cyclamen/enzimologia , Flores/enzimologia , Metiltransferases/metabolismo , Proteínas de Plantas/metabolismo , Antocianinas/química , Cyclamen/genética , Cyclamen/efeitos da radiação , DNA de Plantas/genética , Flores/genética , Flores/efeitos da radiação , Regulação da Expressão Gênica de Plantas/genética , Genoma de Planta/genética , Metilação , Metiltransferases/genética , Metiltransferases/isolamento & purificação , Filogenia , Pigmentação/genética , Proteínas de Plantas/genética , Plantas Geneticamente Modificadas , RNA de Plantas/genética , Análise de Sequência de DNARESUMO
Aberrant hypermethylation of gene promoters is a major mechanism associated with inactivation of tumor suppressor genes (TSGs) in cancer. We have previously shown that the methyl-CpG targeted transcriptional activation (MeTA) that allows re-expression of TSGs in human cancer cells is accomplished by combining a methyl-CpG binding domain (MBD) with a NFkappaB transcriptional activation domain (AD), accompanied by histone H3K9/K14 acetylation. Herein we demonstrate that p300 histone acetyltransferase (HAT), one of the NFkappaB (AD)-associated coactivators, reactivates epigenetically silenced MLH1 in 293T cells. Interestingly, the HAT domain of p300 is not essential for the reactivation of MLH1; instead, the C-terminal transactivation domain (C-TAD) but not the N-terminal one (N-TAD) reactivates MLH1. Furthermore, all ten of the cancer-related genes analyzed in three types of cancer cells were reactivated by the effect of p300 linked to MBD. These results demonstrate that it is possible to reactivate epigenetically silenced TSGs in human cancer cells by direct targeting of a transcriptional coactivator at highly methylated promoters.
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Proteínas Adaptadoras de Transdução de Sinal/genética , Metilação de DNA , Regulação Neoplásica da Expressão Gênica , Genes Supressores de Tumor , Neoplasias/genética , Proteínas Nucleares/genética , Fatores de Transcrição de p300-CBP/metabolismo , Linhagem Celular Tumoral , Ilhas de CpG , Inativação Gênica , Humanos , Proteína 1 Homóloga a MutL , Regiões Promotoras Genéticas , Estrutura Terciária de Proteína , Fatores de Transcrição de p300-CBP/química , Fatores de Transcrição de p300-CBP/genéticaRESUMO
DNA methylation and histone modifications are both major features of the epigenetic silencing seen at tumor suppressor genes (TSGs) in cancer. DNA methylation inhibitors, but not, in general, histone deacetylase, can reactivate TSGs. However, DNA methylation inhibitors frequently upregulate genes whose promoters remain unmethylated. Herein we demonstrated that the methyl-CpG targeted transcriptional activation (MeTA), which allows re-expression of TSGs without DNA demethylation, is widely seen in human cancer. We further analyzed MeTA and found that transcriptional coactivators are recruited at hypermethylated promoter regions of TSGs using the methyl-CpG binding domain (MBD). Reactivation of MLH1 by MeTA accompanied acetylation of histone H3 lysine 9/14 at the promoter region. Furthermore, all ten genes analyzed in three cell lines were reactivated by the effect of MeTA. Our present results lead to an efficient way to search for transcriptionally silenced genes with highly methylated CpG islands, particularly TSGs in cancer and developmentally important genes in embryonic stem cells.
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Ilhas de CpG , Metilação de DNA , Regulação Neoplásica da Expressão Gênica , Inativação Gênica , Genes Supressores de Tumor , Ativação Transcricional , Proteínas Adaptadoras de Transdução de Sinal/genética , Azacitidina/farmacologia , Sequência de Bases , Linhagem Celular Tumoral , Metilação de DNA/efeitos dos fármacos , Desoxicitidina/farmacologia , Humanos , Dados de Sequência Molecular , Proteína 1 Homóloga a MutL , NF-kappa B/metabolismo , Proteínas Nucleares/genética , Análise de Sequência com Séries de Oligonucleotídeos , Regiões Promotoras GenéticasRESUMO
We recently demonstrated that MBD4 possesses the ability to repress transcription through methyl-CpG and is associated with methylated promoters in the CDKN2A and MLH1 genes. In order to further investigate the role of MBD4 in methylation-based transcriptional repression, a yeast two-hybrid screening was performed, and the RET finger protein (RFP) was found to be one of the major proteins that interact with the transcriptional repression domain in MBD4. The effect of the MBD4-mediated transcriptional repression in methylated CDKN2A and MLH1 promoters was extremely enhanced by the overexpression of RFP. Furthermore, RFP forms a protein complex not only with MBD4 but also with MBD2 or MBD3 and was shown to enhance transcriptional repression through MBD2. These results suggest that RFP is a mediator connecting several MBD proteins and allowing the formation of a more potent transcriptional repressor complex. Because RFP has been detected at high levels in a variety of tumor cell lines as well as testis, and embryos, RFP may have an important role in the enhancement of transcriptional repression through MBD proteins in tumorigenesis, spermatogenesis, and embryogenesis.
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Proteínas de Ligação a DNA/metabolismo , Endodesoxirribonucleases/metabolismo , Proteínas Nucleares/metabolismo , Proteínas Repressoras/metabolismo , Transcrição Gênica/fisiologia , Proteínas de Ligação a DNA/genética , Endodesoxirribonucleases/genética , Proteínas Nucleares/genética , Regulação para Cima/fisiologiaRESUMO
We analyzed mutation of the APC, AXIN1, and GSK3genes in 14 pituitary adenomas with abnormal nuclear accumulations of CTNNB1. These tumors did not harbor mutation of the CTNNB1 gene. The genes analyzed encode proteins associated with ubiquitin-mediated degradation of CTNNB1. Although the regions encoding functional domains of these protein products were analyzed, no significant genetic alterations were found. Furthermore, the antibody for the C-terminus of APC detected normal expression of the APC protein in these pituitary adenomas. Our present results imply that an unknown mechanism(s) accelerates the accumulation of CTNNB1 that plays an important role in the pathogenesis of human pituitary adenomas. However, the possibility that mutation of regions outside of our survey or epigenetic mechanism play an important role cannot be excluded.
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Adenoma/genética , Proteína da Polipose Adenomatosa do Colo/genética , Proteínas do Citoesqueleto/metabolismo , Quinase 3 da Glicogênio Sintase/genética , Neoplasias Hipofisárias/genética , Proteínas Repressoras/genética , Transativadores/metabolismo , Adenoma/metabolismo , Adulto , Idoso , Proteína Axina , Análise Mutacional de DNA , Feminino , Regulação Neoplásica da Expressão Gênica , Glicogênio Sintase Quinase 3 beta , Humanos , Peptídeos e Proteínas de Sinalização Intercelular/metabolismo , Masculino , Pessoa de Meia-Idade , Neoplasias Hipofisárias/metabolismo , Transdução de Sinais/genética , Proteínas Wnt , beta CateninaRESUMO
Epigenetic silencing through methyl-CpG (mCpG) is implicated in many biological patterns such as genome imprinting, X chromosome inactivation, and cancer development. In this process, the mCpG binding domain (MBD) proteins play an essential role in transmitting epigenetic information to downstream regulatory proteins. Among the five MBD proteins identified so far, MBD4 has been the only exception; it has long been thought to be a DNA repair protein. Herein we demonstrate that MBD4 has the ability to repress transcription through mCpG. Transcriptional repression by the MBD4 is histone deacetylase (HDAC) dependent, and MBD4 directly binds to Sin3A and HDAC1 at three central regions that overlap transcriptional repression domains. Furthermore, a chromatin immunoprecipitation assay clearly shows that MBD4 binds to hypermethylated promoters of the p16(INK4a) and hMLH1 genes. These results suggest that MBD4 is one of the essential components involved in epigenetic silencing in cancer and its repair activity is necessary for the maintenance of hypermethylated promoters.
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Inibidor p16 de Quinase Dependente de Ciclina/genética , Endodesoxirribonucleases/metabolismo , Epigênese Genética , Proteínas de Neoplasias/genética , Proteínas Nucleares/genética , Proteínas Repressoras/metabolismo , Timina DNA Glicosilase/metabolismo , Proteínas Adaptadoras de Transdução de Sinal , Proteínas de Transporte , Células Cultivadas , Imunoprecipitação da Cromatina , Ilhas de CpG , Metilação de DNA , Endodesoxirribonucleases/genética , Inativação Gênica , Histona Desacetilase 1 , Histona Desacetilases/metabolismo , Humanos , Proteína 1 Homóloga a MutL , Regiões Promotoras Genéticas , Mapeamento de Interação de Proteínas , Proteínas Repressoras/genética , Complexo Correpressor Histona Desacetilase e Sin3 , Timina DNA Glicosilase/genética , Transcrição GênicaRESUMO
Germ-line mutations in the hMLH1 gene are the most frequent cause of hereditary nonpolyposis colorectal cancer and are characterized by missense mutations at high frequency. We found a yeast two-hybrid assay to be an extremely useful and simple tool for evaluating the biological significance of such hMLH1 germ-line missense mutations; 78% (18 of 23) of the missense ones can be recognized as causative for nonpolyposis colorectal cancer. In addition, two of five variants not recognized as causative were thought to be rare polymorphisms. However, we could not detect any differences between wild-type hMLH1 and any of the nine already known polymorphisms causing amino acid alterations. Additional analysis demonstrated that the two-hybrid assay not only detected the dysfunctions at the COOH terminus of the hMLH1 protein necessary for the interaction with associated proteins but also detected a conformational change at the NH(2) terminus carrying ATPase activity. Thus, this method provides a simple and reliable system for accurate diagnosis of hMLH1 alterations.