MiR501-5p may promote proliferation and survival in clear cell kidney carcinoma

MicroRNAs (miRs) are small noncoding RNAs that regulate gene expression and are involved in the control of several biological processes such as proliferation differentiation and apoptosis. The abnormal expression and mutation of miRs is implicated in many hereditary and neoplastic diseases including kidney carcinoma. In the last years, is emerging the necessity to use microRNA as biological biomarkers in order to improve diagnosis, prognosis and therapy response in renal carcinomas. Furthermore, miRNAs might be potential targets for novel therapeutic strategies, especially in patients that are resistant to conventional treatments. We have found that the expression of miR501-5p was higher in 35 clear cell carcinoma (ccRCC) and lower in 9 papillary carcinoma (pRCC) tissues compared with their corresponding normal kidney parenchyma. The expression levels of miR501-5p were not associated with the age of patients or tumor grading in ccRCC and, showed a variable distribution. However, patients with a low expression of miR501-5p exhibited a good prognosis compared with patient with unchanged or high levels of this miR. In order to evaluate the possible role of miR501-5p in renal cancer, we have downregulated and upregulated it by transfection of kidney carcinoma cells (KJ29) with a specific antagomiR and with a plasmid expressing miR501-5p sequences, respectively. The reduction of miR501-5p induced an increase in G0/G1 phase and a decrease of mTOR activity. In addition, the treatment with antagomiR showed an increase in both p53 expression and caspase-3 activity. Consistently, the activation of p53 was demonstrated also by its nuclear translocation. On the contrary, the overexpression of miR501-5p by transfection of KJ29 cells with the plasmid expressing this miR caused the activation of mTOR, the increased expression of MDM2 protein, an inhibitor of p53, and, the reduction of p53 expression. The inhibition of p53 occurred by its degradation through protein ubiquitination and activation of proteasoma machinery which induced a marked increase of cell survival. Taken together, these findings show that the overexpression of miR501-5p may increase cell survival, therefore an high expression of this MicroRNA could be a risk factor for a poor prognosis of renal carcinoma. Conversely, a low expression of miR501-5p seems promote a good prognosis in clear cell kidney carcinoma patients. Thus, the expression of miR501-5p could be used as new potential biomarker for the prognosis of clear cell renal carcinoma

La tipicità del fatto colposo nel diritto penale del lavoro. Tra principi generali in tema di colpa e nuove esigenze di tutela.

The theme of the research project assigned to the writer regards "The peculiarity of the fact culpable in criminal law of labor. Between general principles concerning fault and new protection requirements”. The work is basically divided into three parts: the first part concerning the classical concept of guilt in his profiles philosophical, dogmatic and existential; a second, which concerns the introduction of the precautionary principle in our legal and its impact on the classical concept of guilt; a third part, which concerns the reflection of this principle in the specific area of criminal law of work through an excursus both normative jurisprudential. The present work is, therefore, the deepening of the specific theme consists of the importance of the precautionary principle in contexts that involve variously safety and reflections of the same in the specific area of criminal law of the work with reference to the evolution of the classical concept of criminal guilt. The research carried out, therefore, in the first phase was concerned with the study of the classical concept of guilt. Then I analyzed the different conceptions of guilt developed in doctrine and the basic elements of the judgment negligent liability, namely the unwillingness of the fact, the precautionary rules, the c.d. concretization of risk, predictability and avoidance detrimental event and, finally, the enforceability by the party's agent alternative behavior lawful. In a second step was carried out, then, the study of the c.d. precautionary principle as a criterion of risk management in terms of scientific uncertainty. On the theme, on the one hand, sources, assumptions and fields of application of that principle were analyzed, the other the legitimacy of that under criminal law, identifying the differences between the criminal law of prevention and precaution, from 'more deformations that following the entry of this principle in our system are undergoing causation and guilt criminal with particular reference to the evolution of the concept of predictability of the event. On the basis of the deepening made you come to the conclusion that this principle, pending the generic prescriptive that characterizes it, is likely to conflict with the character of certainty of the criminal standard and lends itself to play a role as a potential factor in the expansion of the categories of classical criminal law. It could, in fact, affect two fundamental factors for the configuration of the typical fact: first, on the objective level of causation, exploiting the probabilistic structure and turning it into a "nexus of risk"; secondly on the subjective level of guilt deforming in its more recognizable cognitive risk and predictability of the result. Finally, the third part of the research project has been directed towards the study of reflexes that the precautionary principle has had in the specific field of criminal law of work through an excursus both normative jurisprudential. The matter of safety at work, the expected legal interests protected by primary degree, you are, indeed, characterizing a transition from rule to rule precautionary precautionary: that fact marks the release of preventing the event from the conduct conforms to the rule previously identified by placing, consequently, in the crisis the same function garantistic rule precautionary. In this sector, indeed, one might come to affirm the responsibility culpable of a subject matter a concrete verification about the actual effectiveness of the predictive rule remand in relation to the specific offense to the legal interest protected by the rule, so as to impute the event even where there is only the possibility that the detrimental effects occur, with direct and obvious impact on the subjective level of predictability and avoidance, which are no longer subject to a specific event, but rather a class or kind of events. Emblematic on the subject, and object of study, some judgments regarding the exposure of workers to asbestos or harmful substances such as vinyl chloride monomer. In response, in fact, the protection of victims and the needs of substantial justice, we are witnessing an obvious flexibility of the paradigm of guilt and criminal liability from asbestos, offering a rich case law, constitutes a particular field of reflection for what concerns the evolution of the classical dogmatic categories of criminal law for charging the damaging event. Jurisprudential analysis performed, it seems, indeed, take off a micro-penal system from occupational exposures, where the deconstruction of guilt, through the distortion of the value of the regulatory rule precautionary determines an instance of absolute protection of the victim, with a corresponding load iperdeterrece for the author, called to bear risks criminal gravitating outside its sphere of recognition

Diagnostico e monitorização da infecção ativa por citomegalovirus em transplantados alogenicos de celulas progenitoras hematopoeticas

Orientador: Sandra Cecilia Botelho CostaTese (doutorado) - Universidade Estadual de Campinas, Faculdade de Ciencias MedicasResumo: Neste estudo, monitorizamos 69 pacientes receptores de medula óssea e de células progenitoras periféricas, alogênicos, HLA idênticos, desde o pré-transplante até 150 dias após o transplante, semanalmente com as técnicas de PCR dupla e antigenemia e mensalmente com sorologia tipo ELISA (IgM e IgG) contra o HCMV, incluindo os doadores no pré-transplante. Quarenta e cinco pacientes do Grupo A, sendo 38 adultos e 7 crianças, que possuíam doenças hematológicas malignas e status sorológico de risco, receberam profilaxia universal com ganciclovir; e vinte e quatro pacientes do Grupo B, sendo 14 adultos com anemia aplástica e 10 crianças (7 com doenças hematológicas malignas e 3 com anemia aplástica) não receberam profilaxia com ganciclovir. Se a vigilância laboratorial era positiva (2 ou mais amostras de PCR positivas consecutivas ou 1 ou mais células antígeno-positivas por antigenemia), terapia precoce com ganciclovir era administrada, seguida de doses de manutenção nos pacientes de ambos os grupos. Como resultados obtidos após a monitorização, nos pacientes do Grupo A, 36/45 pacientes tiveram infecção ativa pelo HCMV detectada por AGM e/ou PCR. A probabilidade de antigenemia positiva até o dia +150 foi de 53% e por PCR foi de 71,3% (P=0,04). O teste de PCR não se mostrou mais precoce na detecção da infecção ativa pelo HCMV do que a antigenemia nos pacientes deste grupo (54 e 62 dias, respectivamente, P=NS). A sensibilidade e a especificidade da antigenemia foi de 58% e 64,3%, respectivamente, usando a PCR como padrão-ouro. Nos pacientes do Grupo B, 19/24 pacientes tiveram infecção ativa pelo HCMV detectada por AGM e/ou PCR. A probabilidade de positivação da antigenemia foi de 66,7% e da PCR dupla foi de 67,2% (P=NS). O teste de PCR mostrou-se mais precoce do que a antigenemia na detecção da Infecção Ativa por HCMV nos pacientes do Grupo B (24 e 34 dias, respectivamente, P=0,03). A sensibilidade e a especificidade da antigenemia foi de 81,2% e 62,5%, respectivamente, usando a PCR dupla como padrão-ouro. Doença por HCMV ocorreu em 2/45 pacientes (4,44%) do Grupo A (casos de gastrite), e foram tratados com sucesso pelo ganciclovir. No Grupo B, 4/24 pacientes (16,7%) tiveram doença por HCMV. Um desses pacientes teve coriorretinite, dois tiveram gastrite e um teve pneumonia intersticial. Dois óbitos por HCMV ocorreram neste grupo (50%), sendo que um por gastrite inicial e outro por pneumonia intersticial, ambos com disseminação sistêmica posterior. Neste trabalho, o uso da profilaxia universal com ganciclovir pareceu retardar o aparecimento da infecção ativa, pois atrasou a positividade dos testes, quando comparado ao grupo que não recebeu profilaxia. Contudo, nenhum impacto favorável foi observado no grupo de pacientes que recebeu profilaxia universal com ganciclovir, em relação à infecção ativa pelo HCMV, doença e sobrevida. Outra observação foi em relação ao comportamento dos testes diagnósticos nos pacientes tratados com ganciclovir, onde a PCR dupla demonstrou-se mais precoce, porém, na maioria dos casos, a antigenemia confirmou o diagnóstico. Nossos resultados sugerem que tanto a antigenemia quanto a PCR podem ser utilizadas como marcadores precoces para a introdução da terapia antiviral específicaAbstract: Human cytomegalovirus (HCMV) remains a cause of significant morbidity and mortality after allogeneic hematopoietic stem cell transplantation (HSCT). Different prophylactic approaches have been adopted, but these strategies remain controversial. Early detection of active HCMV infection is imperative to control and to detain HCMV disease. Ganciclovir universal prophylaxis or pre-emptive treatment based on detection of antigenemia (AGM) or a nested polymerase chain reaction (N-PCR) effectively prevents HCMV disease during the first 100 days after transplant. The aim of this study was to describe our experience in the control of active HCMV infection using ganciclovir universal prophylaxis at low doses and pre-emptive therapy with ganciclovir following hematopoietic stem cell transplantation (HSCT) based on the monitoring of antigenemia (AGM) and on a nested polymerase chain reaction (N-PCR). We prospectively monitored 69 HLA identical sibling donor HSCT patients for evidence of HCMV infection and disease starting in the day of transplant until day 150 after transplant. Two groups of patients were studied: Group A: 45 recipients with malignances diseases and with risk factor for HCMV disease (D+/R+; D-/R+ and D+/R-) received ganciclovir universal prophylaxis in a dose of 5 mg/kg per day, 3 days per week and continued until day 75 after transplant; Group B: 14 adults patients, most of them with non-malignance disease and ten pediatrics patients who did not receive ganciclovir universal prophylaxis. In Group A, the incidence of positive antigenemia at day 150 was 51% in a median of 62 days after transplantation, with a positive PCR of 68.9% in a median of 54 days. In Group B, the antigenemia positivity was 66.6% and that of N-PCR was 66.7% in a median of 34 and 24 days, respectively. HCMV disease occurred in 6/55 patients (10.9%), with 2/36 (5.5%) from Group A (gastritis) and was successfully treated with ganciclovir. In Group B, 4/24 patients (16.7%) had HCMV disease (one chorioretinitis; two gastritis and one with interstitial pneumonia (IP)). Two of these patients (50%) died of HCMV disease. Our results suggest that antigenemia and N-PCR can be used as markers for assessing the introduction pre-emptive therapy. This approach could prove to be more cost-effective than ganciclovir universal prophylaxis for treating HCMV infectionDoutoradoCiencias BasicasDoutor em Clínica Médic

Evaluation Of An In-house Specific Immunoglobulin G (igg) Avidity Elisa For Distinguishing Recent Primary From Long-term Human Cytomegalovirus (hcmv) Infection.

This article describes the standardization and evaluation of an in-house specific IgG avidity ELISA for distinguishing recent primary from long-term human cytomegalovirus (HCMV) infection. The test was standardized with the commercial kit ETI-CYTOK G Plus (Sorin Biomedica, Italy) using 8 M urea in phosphate-buffered saline to dissociate low-avidity antibodies after the antigen-antibody interaction. The performance of the in-house assay was compared to that of the commercial automated VIDAS CMV IgG avidity test (bioM rieux, France). Forty-nine sera, 24 from patients with a recent primary HCMV infection and 25 from patients with a long-term HCMV infection and a sustained persistence of specific IgM antibodies, were tested. Similar results were obtained with the two avidity methods. All 24 sera from patients with recently acquired infection had avidity indices compatible with acute HCMV infection by the VIDAS method, whereas with the in-house method, one serum sample had an equivocal result. In the 25 sera from patients with long-term infection, identical results were obtained with the two methods, with only one serum sample having an incompatible value. These findings suggest that our in-house avidity test could be a potentially useful tool for the immunodiagnosis of HCMV infection.45323-

Avaliação de um teste de avidez imunoenzimático para o citomegalovírus humano (ELISA-HCMV) para distinguir a infecção primária recente da infecção de longa duração

This article describes the standardization and evaluation of an in-house specific IgG avidity ELISA for distinguishing recent primary from long-term human cytomegalovirus (HCMV) infection. The test was standardized with the commercial kit ETI-CYTOK G Plus (Sorin Biomedica, Italy) using 8 M urea in phosphate-buffered saline to dissociate low-avidity antibodies after the antigen-antibody interaction. The performance of the in-house assay was compared to that of the commercial automated VIDAS CMV IgG avidity test (bioMérieux, France). Forty-nine sera, 24 from patients with a recent primary HCMV infection and 25 from patients with a long-term HCMV infection and a sustained persistence of specific IgM antibodies, were tested. Similar results were obtained with the two avidity methods. All 24 sera from patients with recently acquired infection had avidity indices compatible with acute HCMV infection by the VIDAS method, whereas with the in-house method, one serum sample had an equivocal result. In the 25 sera from patients with long-term infection, identical results were obtained with the two methods, with only one serum sample having an incompatible value. These findings suggest that our in-house avidity test could be a potentially useful tool for the immunodiagnosis of HCMV infection.Este artigo descreve a padronização e avaliação de um teste de avidez imunoenzimático para o citomegalovírus humano (ELISA-HCMV) para distinguir a infecção primária recente da infecção de longa duração. O teste foi padronizado com o kit comercial ETI-CYTOK G Plus (Sorin Biomedica, Itália), utilizando uréia 8 M para a dissociação dos anticorpos de baixa avidez. A performance do teste ELISA-HCMV foi comparada com a do teste de avidez comercial automatizado VIDAS CMV IgG (bioMérieux, França), utilizando 24 soros de pacientes com infecção primária recente e 25 soros de pacientes com infecção de longa duração apresentando persistência de anticorpos específicos IgM. Resultados similares foram obtidos com os dois métodos de avidez. Todos os 24 soros de pacientes com infecção recentemente adquirida apresentaram índices de avidez compatíveis com infecção aguda pelo HCMV utilizando o teste VIDAS CMV IgG, enquanto que um dos soros apresentou resultado duvidoso no teste ELISA-HCMV. Os 25 soros de pacientes com infecção de longa duração apresentaram resultados idênticos com os dois métodos, com apenas um dos soros apresentando um valor não compatível. Estes resultados sugerem que o teste de avidez descrito pode ser potencialmente útil para o imunodiagnóstico da infecção pelo HCMV

Estudo comparativo entre sorologia, antigenemia e reação em cadeia da polimerase para o monitoramento da infecção por citomegalovírus em pacientes receptores de transplantes de células progenitoras hematopoéticas

Forty-six allogeneic hematopoietic stem cell transplantation (HSCT) patients were monitored for the presence of CMV antibodies, CMV-DNA and CMV antigens after transplantation. Immunoenzymatic serological tests were used to detect IgM and the increase in CMV IgG antibodies (increase IgG), a nested polymerase chain reaction (N-PCR) was used to detect CMV-DNA, and an antigenemia assay (AGM) was used to detect CMV antigens. The presence of CMV-IgM and/or CMV-increase IgG antibodies was detected in 12/46 (26.1%) patients, with a median time between HSCT and the detection of positive serology of 81.5 days. A positive AGM was detected in 24/46 (52.2%) patients, with a median time between HSCT and antigen detection of 62 days. Two or more consecutive positive N-PCR results were detected in 32/46 (69.5%) patients, with a median time between HSCT and the first positive PCR of 50.5 days. These results confirmed that AGM and mainly PCR are superior to serology for the early diagnosis of CMV infection. Six patients had CMV-IgM and/or CMV-increase IgG with a negative AGM (five cases) or N-PCR assay (one case). In five of these cases the serological markers were detected during the first 100 days after HSCT, the period of highest risk. These findings support the idea that serology may be useful for monitoring CMV infections in HSCT patients, especially when PCR is unavailable.Quarenta e seis pacientes receptores de transplantes de células progenitoras hematopoéticas (TCPH) foram monitorados em relação à infecção ativa por citomegalovírus (CMV). Testes sorológicos imunoenzimáticos foram utilizados para a detecção de anticorpos IgM e elevação significativa das concentrações de anticorpos IgG (aumento IgG), nested-PCR (N-PCR) foi utilizada para a detecção de CMV-DNA e antigenemia (AGM) para a detecção de antígenos virais. A presença de CMV-IgM e/ou CMV-aumento IgG foi detectada em 12/46 (26,1%) pacientes, sendo o tempo mediano entre o transplante e a detecção dos marcadores sorológicos de 81,5 dias; AGM positiva foi detectada em 24/46 (52,2%) pacientes, sendo o tempo mediano entre o transplante e a detecção de antígenos virais de 62 dias. Dois ou mais resultados positivos consecutivos de N-PCR foram detectados em 32/46 (69,5%) pacientes, sendo o tempo mediano entre o transplante e o primeiro teste positivo de 50,5 dias. Esses resultados confirmaram que a AGM e principalmente a PCR são superiores à sorologia, com relação ao diagnóstico da infecção pelo CMV. Seis pacientes apresentaram reações CMV-IgM positivas e/ou CMV-aumento IgG com reações negativas de AGM (cinco casos) ou N-PCR (um caso). Em cinco desses casos, os marcadores sorológicos foram detectados nos 100 primeiros dias após o transplante, considerado o período de maior risco. Esses resultados indicam que os testes sorológicos podem ser úteis no monitoramento da infecção por CMV após o transplante de células progenitoras hematopoéticas, principalmente quando a N-PCR não for disponível

The prevalence of adenoviral conjunctivitis at the Clinical Hospital of the State University of Campinas, Brazil

Viral conjunctivitis is a common, highly contagious disease that is often caused by an adenovirus. The aim of this study was to evaluate the prevalence of adenoviral conjunctivitis by analyzing data from a prospective clinical study of 122 consecutively enrolled patients who were treated at the Clinical Hospital of the State University of Campinas (UNICAMP) after a clinical diagnosis of infectious conjunctivitis between November 2011 and June 2012. METHODS: Polymerase chain reaction was used to evaluate all cases of clinically diagnosed infectious conjunctivitis and based on the laboratory findings, the prevalence of adenoviral infections was determined. The incidence of subepithelial corneal infiltrates was also investigated. RESULTS: Of the 122 patients with acute infectious conjunctivitis included, 72 had positive polymerase chain reaction results for adenoviruses and 17 patients developed subepithelial corneal infiltrates (13.93%). CONCLUSIONS: The polymerase chain reaction revealed that the prevalence of adenoviral conjunctivitis was 59% in all patients who presented with a clinical diagnosis of infectious conjunctivitis from November 2011 to June 2012. The prevalence of adenoviral conjunctivitis in the study population was similar to its prevalence in other regions of the world701174875

The prevalence of adenoviral conjunctivitis at the Clinical Hospital of the State University of Campinas, Brazil

OBJECTIVES: Viral conjunctivitis is a common, highly contagious disease that is often caused by an adenovirus. The aim of this study was to evaluate the prevalence of adenoviral conjunctivitis by analyzing data from a prospective clinical study of 122 consecutively enrolled patients who were treated at the Clinical Hospital of the State University of Campinas (UNICAMP) after a clinical diagnosis of infectious conjunctivitis between November 2011 and June 2012. METHODS: Polymerase chain reaction was used to evaluate all cases of clinically diagnosed infectious conjunctivitis and based on the laboratory findings, the prevalence of adenoviral infections was determined. The incidence of subepithelial corneal infiltrates was also investigated. RESULTS: Of the 122 patients with acute infectious conjunctivitis included, 72 had positive polymerase chain reaction results for adenoviruses and 17 patients developed subepithelial corneal infiltrates (13.93%). CONCLUSIONS: The polymerase chain reaction revealed that the prevalence of adenoviral conjunctivitis was 59% in all patients who presented with a clinical diagnosis of infectious conjunctivitis from November 2011 to June 2012. The prevalence of adenoviral conjunctivitis in the study population was similar to its prevalence in other regions of the world

Active human Cytomegalovirus infection and Glycoprotein B genotypes in Brazilian pediatric renal or hematopoietic stem cell transplantation patients

A prospective analysis of active Human Cytomegalovirus infection (HCMV) was conducted on 33 pediatric renal or hematopoietic stem cell post-transplant patients. The HCMV-DNA positive samples were evaluated for the prevalence of different gB subtypes and their subsequent correlation with clinical signs. The surveillance of HCMV active infection was based on the monitoring of antigenemia (AGM) and on a nested polymerase chain reaction (N-PCR) for the detection of HCMV in the patients studied. Using restriction analysis of the gB gene sequence by PCR-RFLP (Restriction Fragment Length Polymorphism), different HCMV strains could be detected and classified in at least four HCMV genotypes. Thirty-three pediatric recipients of renal or bone marrow transplantation were monitored. Twenty out of thirty-three (60.6%) patients demonstrated active HCMV infection. gB1 and gB2 genotypes were more frequent in this population. In this study, we observed that gB2 had correlation with reactivation of HCMV infection and that patients with mixture of genotypes did not show any symptoms of HCMV disease. Future studies has been made to confirm this.505