Donor-derived TB after kidney transplantation: a case report / TB derivada do doador após transplante renal: um relato de caso

Abstract Introduction: Tuberculosis (TB) is a possible serious complication of solid organ transplantation, associated with high mortality and morbidity. Post-transplant TB has varied pathogenesis with many approaches to its prevention, which is the most important way to reduce its incidence. Treatment of TB in organ recipients is challenging because of drug toxicity and interaction with immunosuppressants. Case report: an 18-year-old woman that underwent kidney transplantation from a deceased donor and was discharged with fair renal function was readmitted at 37th postoperative day with fever. CT showed signs of miliary TB and fluid collection besides graft fistulization through the skin. The patient presented positive BAAR in the drained fluid and Koch's bacillus in the urine. She was treated with a four-drug regimen (rifampicin, isoniazid, pyrazinamide, and etambutol), with great response and preserved graft function. We were informed that the recipient of the contralateral kidney also presented post-transplant TB, implying in a donor-derived origin. Conclusion: TB is an important differential diagnosis for infectious complications in patients after solid-organ transplantation, especially in endemic regions. Its initial clinical presentation can be unspecific and it should be suspected in the presence of fever or formation of fluid collections. The suspicion of TB is the key to early diagnosis and satisfactory outcomes in post-transplant TB.

Resumo Introdução: A tuberculose (TB) é uma possível complicação grave do transplante de órgãos sólidos, associada à alta mortalidade e morbidade. A TB pós-transplante tem patogênese variada com muitas abordagens para sua prevenção, que é a forma mais importante de reduzir sua incidência. O tratamento da TB em receptores de órgãos é um desafio devido à toxicidade dos medicamentos e à interação com imunossupressores. Relato de caso: uma mulher de 18 anos que foi submetida a transplante renal de um doador falecido e recebeu alta com função renal adequada foi readmitida no 37º dia de pós-operatório com febre. A TC mostrou sinais de TB miliar e coleção de fluidos além de fistulização do enxerto através da pele. A paciente apresentou BAAR positivo no fluido drenado e bacilo de Koch na urina. Ela foi tratada com um esquema de quatro medicamentos (rifampicina, isoniazida, pirazinamida e etambutol), com ótima resposta e função de enxerto preservada. Fomos informados de que o receptor do rim contralateral também apresentou TB pós-transplante, implicando em uma origem derivada do doador. Conclusão: A TB é um importante diagnóstico diferencial para complicações infecciosas em pacientes após transplante de órgãos sólidos, especialmente em regiões endêmicas. Sua apresentação clínica inicial pode não ser específica e deve ser suspeitada na presença de febre ou formação de coleções de fluidos. A suspeita de TB é a chave para o diagnóstico precoce e desfechos satisfatórios na TB pós-transplante.

Resumen ejecutivo: Enfermedad por hígado graso no alcohólico en sujetos con diabetes mellitus tipo 2: Postura conjunta de la Asociación Chilena de Hepatología (ACHHEP) y la Sociedad Chilena de Diabetología (SOCHIDIAB) / Non-alcoholic fatty liver disease in type 2 diabetes: a joint statement of the Chilean Hepatology and Diabetology Societies

Non-alcoholic fatty liver disease (NAFLD) has a high prevalence and risk of progression to cirrhosis and other complications in patients with type 2 diabetes mellitus (T2DM). Likewise, the presence of NAFLD implies a high risk of developing T2DM, determining a bidirectional relationship between them. The diabetology and hepatology societies, developed a joint initiative aiming to unify criteria, reviewing the definitions, diagnostic criteria, risk stratification, treatment, and follow-up of patients with NAFLD and T2DM. The key questions to be discussed were defined by a panel of specialists in diabetology and hepatology. The Delphi methodology was used to reach consensus on the respective recommendations. Based on the discussion generated among the experts, diagnostic and treatment algorithms were proposed, as well as an indication for referral and the role of the different specialists involved in the management of these patients. Strengthening multidisciplinary work with patients with NAFLD and T2DM will allow the early recognition of the disease, the prevention of the progression to cirrhosis, and reducing the associated complications.

Bothrops moojeni L-amino acid oxidase induces apoptosis and epigenetic modulation on Bcr-Abl+ cells

Resistance to apoptosis in chronic myeloid leukemia (CML) is associated with constitutive tyrosine kinase activity of the Bcr-Abl oncoprotein. The deregulated expression of apoptosis-related genes and alteration in epigenetic machinery may also contribute to apoptosis resistance in CML. Tyrosine kinase inhibitors target the Bcr-Abl oncoprotein and are used in CML treatment. The resistance of CML patients to tyrosine kinase inhibitors has guided the search for new compounds that may induce apoptosis in Bcr-Abl+ leukemic cells and improve the disease treatment. Methods: In the present study, we investigated whether the L-amino acid oxidase isolated from Bothrops moojeni snake venom (BmooLAAO-I) (i) was cytotoxic to Bcr-Abl+ cell lines (HL-60.Bcr-Abl, K562-S, and K562-R), HL-60 (acute promyelocytic leukemia) cells, the non-tumor cell line HEK-293, and peripheral blood mononuclear cells (PBMC); and (ii) affected epigenetic mechanisms, including DNA methylation and microRNAs expression in vitro. Results: BmooLAAO-I induced ROS production, apoptosis, and differential DNA methylation pattern of regulatory apoptosis genes. The toxin upregulated expression of the pro-apoptotic genes BID and FADD and downregulated DFFA expression in leukemic cell lines, as well as increased miR-16 expression - whose major predicted target is the anti-apoptotic gene BCL2 - in Bcr-Abl+ cells. Conclusion: BmooLAAO-I exerts selective antitumor action mediated by H2O2 release and induces apoptosis, and alterations in epigenetic mechanisms. These results support future investigations on the effect of BmooLAAO-I on in vivo models to determine its potential in CML therapy.(AU)

Mucosite oral em pacientes pediátricos submetidos a transplante de células hematopoiéticas / Oral mucositis in pediatric patients undergoing hematopoietic cell transplantation

A mucosite oral (MO) constitui uma das principais morbidades nos períodos imediatos após o transplante de células hematopoiéticas (TCH). Em pacientes pediátricos, a MO é pouco descrita na literatura, não havendo protocolos específicos para prevenção e tratamento dessas lesões. O objetivo deste estudo foi caracterizar a MO em pacientes pediátricos antes e durante os períodos recentes após o TCH, bem como verificar quais fatores relacionados ao paciente e ao transplante podem estar associados ao tempo de duração e à gravidade dessas lesões. Foi também analisado o impacto da MO nas alterações de massa corpórea, na prescrição de opióides e nutrição artificial, bem como na sobrevida global. Para tanto, foram selecionados 115 prontuários de pacientes com idade igual ou menor que 18 anos, que foram submetidos a TCH, coletando-se informações sobre o paciente, o transplante e a toxicidade no trato gastrointestinal, desde o período de condicionamento até 10 dias após a pega da medula. Os pacientes foram divididos nas faixas etárias de 0 a 2, 3 a 6, 7 a 12, e 13 a 18 anos. Observou-se que houve maior frequência de pacientes com indicação de TCH devido a imunodeficiências primárias (34.78%), seguido de pacientes portadores de neoplasias hematológicas (24.34%) e anemias (18.26%). Houve alta frequência de pacientes na faixa etária de 0 a 2 anos (n=35 pacientes) e de 7 a 12 anos (n=38). Esses pacientes foram submetidos principalmente a transplante alogênico (87.83%) e condicionamento contendo bussulfano (57.40%). Observou-se maior frequência de MO graus 3 e 4 nos pacientes na faixa etária de 7 a 12 anos em comparação às demais faixas etárias (26.32%, p=0.019). Maior tempo de duração de dor para deglutir também foi observado nessa faixa etária e na de 13 a 18 anos (p=0.024). Em geral, em todas as idades, a MO não ficou restrita ao período de neutropenia, manifestando-se antes ou estendendo-se além desse período. A prescrição de nutrição artificial (71.30%) e de opióides (84.16%) foi alta na amostra como um todo. Os pacientes com 0 a 2 anos de idade necessitaram de nutrição artificial por mais tempo (p=0.002). A profilaxia para doença do enxerto contra o hospedeiro (DECH) com metotrexato foi fator de risco significativo para MO. Tanto a MO grau³2 (OR=12.6, p=0.017) quanto a duração de dor para deglutir ³3 dias (OR=6.5, p<0.001) impactaram significativamente na prescrição de opióides. A dor para deglutir também impactou na prescrição de nutrição artificial (OR=2.7, p=0.018), mas a MO não. Nenhum fator relacionado à mucosite no trato digestivo teve associação significativa com perda ou ganho de massa corpórea, nem com sobrevida global. Concluiu-se que os pacientes pediátricos que foram submetidos a TCH exibiram perfis de MO distintos segundo a faixa etária, com variações no período de manifestação, na gravidade e na duração dessas lesões. Protocolos específicos de cuidados orais para essas faixas etárias devem ser instituídos, focados principalmente no controle da MO no tocante a analgesia.

BthTX-I from Bothrops jararacussu induces apoptosis in human breast cancer cell lines and decreases cancer stem cell subpopulation

Breast cancer is the neoplasm with both the highest incidence and mortality rate among women worldwide. Given the known snake venom cytotoxicity towards several tumor types, we evaluated the effects of BthTX-I from Bothrops jararacussu on MCF7, SKBR3, and MDAMB231 breast cancer cell lines. Methods: BthTX-I cytotoxicity was determined via MTT 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazoliumbromide assay. Cell death was measured by a hypotonic fluorescent solution method, annexin-V-FITC/propidium iodide staining and by apoptotic/autophagic protein expression. Cancer stem cells (CSCs) were quantified by flow cytometry using anti-CD24-FITC and anti-CD44-APC antibodies and propidium iodide. Results: BthTX-I at 102 µg/mL induced cell death in all cell lines. The toxin induced apoptosis in MCF7, SKBR3, and MDAMB231 in a dose-dependent manner, as confirmed by the increasing number of hypodiploid nuclei. Expression of pro-caspase 3, pro-caspase 8 and Beclin-1 proteins were increased, while the level of the antiapoptotic protein Bcl-2 was diminished in MCF7 cells. BthTX-I changed the staining pattern of CSCs in MDAMB231 cells by increasing expression of CD24 receptors, which mediated cell death. Conclusions: BthTX-I induces apoptosis and autophagy in all breast cancer cell lines tested and also reduces CSCs subpopulation, which makes it a promising therapeutic alternative for breast cancer.(AU)

Philadelphia-negative myeloproliferative neoplasms as disorders marked by cytokine modulation

ABSTRACT Background: Cytokines are key immune mediators in physiological and disease processes, whose increased levels have been associated with the physiopathology of hematopoietic malignancies, such as myeloproliferative neoplasms. Methods: This study examined the plasma cytokine profiles of patients with essential thrombocythemia, primary myelofibrosis, polycythemia vera and of healthy subjects, and analyzed correlations with JAK2 V617F status and clinical-hematological parameters. Results: The proinflammatory cytokine levels were increased in myeloproliferative neoplasm patients, and the presence of the JAK2 V617F mutation was associated with high IP-10 levels in primary myelofibrosis patients. Conclusions: Essential thrombocythemia, primary myelofibrosis, and polycythemia vera patients exhibited different patterns of cytokine production, as revealed by cytokine network correlations. Together, these findings suggest that augmented cytokine levels are associated with the physiopathology of myeloproliferative neoplasms.

Cytotoxic and pro-apoptotic action of MjTX-I, a phospholipase A2 isolated from Bothrops moojeni snake venom, towards leukemic cells

A leucemia mieloide crônica (LMC) é uma neoplasia mieloproliferativa BCR-ABL1 + marcada por aumento da mieloproliferação e presença de células leucêmicas resistentes à apoptose. A terapia de primeira linha atual para a LMC é a administração de inibidores da tirosina quinase, mesilato de imatinibe, dasatinibe ou nilotinibe. Embora eficaz no tratamento da LMC, alguns pacientes se tornaram resistentes a essa terapia, levando à progressão da doença e à morte. Assim, a descoberta de novos compostos para melhorar a terapia da LMC ainda é um desafio. Aqui, os destinatários se MjTX-I, uma fosfolipase A 2 isolado a partir de Bothrops moojeni de veneno de cobra, afecta a viabilidade de Bcr-Abl de mesilato de imatinib-resistente + linhas celulares. Métodos: Examinamos o efeito citotóxico e pró-apoptótico de MjTX-I em células K562-S e K562-R Bcr-Abl + e na linha de células HEK-293 não tumorais e células mononucleares de sangue periférico, usando o 3- (4, Brometo de 5-dimetiltiazol-2-il) -2,5-difeniltetrazólio e os métodos de solução fluorescente hipotônica, associados à detecção de ativação de caspases 3, 8 e 9 e clivagem de poli (ADP-ribose) polimerase (PARP). Também analisamos o potencial MjTX-I para modular a expressão de genes relacionados à apoptose em células K562-S e K562-R. Resultados: O MjTX-I diminuiu a viabilidade das células K562-S e K562-R em 60 a 65%, sem afetar a viabilidade das células não tumorais, ou seja, exerceu citotoxicidade seletiva para as linhagens celulares Bcr-Abl + . Em linhas de células leucêmicas, a toxina induziu apoptose, caspases 3, 8 e 9 ativadas, PARP clivada, expressão negativa do gene anti-apoptótico BCL-2 e expressão aumentada do gene pró-apoptótico BAD. Conclusão: O efeito antitumoral de MjTX-I está associado ao seu potencial para induzir apoptose e citotoxicidade em linhagens celulares positivas para Bcr-Abl sensíveis e resistentes ao mesilato de imatinibe, indicando que MjTX-I é um candidato promissor a fármaco para atualizar a terapia de LMC.(AU)

l-Amino acid oxidase isolated from calloselasma rhodostoma snake venom induces cytotoxicity and apoptosis in JAK2V617F-positive cell lines

BACKGROUND: Myeloproliferative neoplasms are Philadelphia chromosome-negative diseases characterized by hyperproliferation of mature myeloid cells, associated or not with the Janus kinase 2 tyrosine kinase mutation, JAK2V617F. As there is no curative therapy, researchers have been investigating new drugs to treat myeloproliferative neoplasms, including l-amino acid oxidase from Calloselasma rhodostoma snake venom (CR-LAAO), which is a toxin capable of eliciting apoptosis in several tumor cell lines. OBJECTIVE: To evaluate the effects of l-amino acid oxidase from C. rhodostoma snake venom in the apoptotic machinery of JAK2-mutated cell lines. METHODS: The HEL 92.1.7 and SET-2 cell lines were cultured with l-amino acid oxidase and catalase for 12 h at 37 °C in 5% carbon dioxide. The cell viability was assessed by the multi-table tournament method, the level of apoptosis was measured by flow cytometry, and the expression of cysteine-dependent aspartate-specific proteases and cleaved Poly(ADP-ribose) polymerase were analyzed by Western blotting. RESULTS: l-Amino acid oxidase from C. rhodostoma snake venom was cytotoxic to HEL 92.1.7 and SET-2 cells (50% inhibitory concentration = 0.15 µg/mL and 1.5 µg/mL, respectively) and induced apoptosis in a concentration-dependent manner. Cell treatment with catalase mitigated the l-amino acid oxidase toxicity, indicating that hydrogen peroxide is a key component of its cytotoxic effect.The activated caspases 3 and 8 expression and cleaved PARP in HEL 92.1.7 and SET-2 cells confirmed the apoptosis activation by CR-LAAO. CONCLUSIONS: l-Amino acid oxidase from C. rhodostoma snake venom is a potential antineoplastic agent against HEL 92.1.7 and SET-2 JAK2V617F-positive cells as it activates the extrinsic apoptosis pathway.

Neumonía lipoidea tras aspiración accidental de vaselina / Lipoid pneumonia after accidental aspiration of vaseline

La neumonía lipoidea es una patología pulmonar poco conocida que resulta de la acumulación de lípidos de origen endógenos o exógenos a nivel alveolar pulmonar. Suele ser subdiagnosticada dada que la presentación clínica es inespecífica, por lo que suele confundirse con otras patologías broncopulmonares, muchas veces no llegándose al diagnóstico preciso de esta. Dentro de los datos anamnesticos el antecedente, ya sea de uso reciente o de larga data, de la ingesta y/o inhalación accidental o voluntaria de alguna sustancia que contenga algún componente lipídico en su base es primordial. Pacientes de edades extremas suelen ser los más afectados y en quienes debemos tener la sospecha diagnóstica. Su diagnóstico es complejo requiriendo historia clínica, imágenes que apoyen un compromiso pulmonar (de preferencia tomografía computada) e idealmente la detección intraalveolar de lípidos y macrófagos cargados de éstos en el lavado broncopulmonar. Actualmente, no existe consenso sobre el manejo específico de esta patología. Lo importante es identificar la causa y descontinuar su uso. En neumonía lipoidea de causa exógena se recomienda manejo de soporte según sea la presentación clínica del paciente.

Lipoid pneumonia (LN) is an unknown lung disease due to lipid accumulation, of endogenous or exogenous origin, at the pulmonary alveolar level. Often it is underdiagnosed because of its nonespecific presentation whose differential diagnosis are other bronchopulmonary diseases. Anamnestic data, either recent or long-standing use of the intake and / or accidental or deliberate inhalationof any substance containing a lipid component as its base is relevant information. Extreme age groups are the most affected, and in whom wemust suspect LN diagnosis. The diagnosis of LN is complex and requires a complete medical history, images that support lung involvement (preferably computed tomography), and ideally intraalveolar detection of lipid-ladenmacrophages in the bronchoalveolar lavage. Currently, there is no consensus on the specific management of this disease. It is important to identify the cause and discontinue its exposure. Whatever the varity of the clinical presentation, support magement is recommended to treat LN.

Bothrops snake venoms and their isolated toxins, an L-amino acid oxidase and a serine protease, modulate human complement system pathways

Background Activation of the complement system plays an important role in the regulation of immune and inflammatory reactions, and contributes to inflammatory responses triggered by envenomation provoked byBothrops snakes. The present study aimed to assess whether Bothrops jararacussuand Bothrops pirajai crude venoms and their isolated toxins, namely serine protease (BjussuSP-I) and L-amino acid oxidase (BpirLAAO-I), modulate human complement system pathways.Methods Lyophilized venom and toxin samples solubilized in phosphate buffered saline were diluted in appropriate buffers to evaluate their hemolytic activity on the alternative and classical pathways of the complement system. Venom- and toxin-treated normal human serum was added to the erythrocyte suspension, and the kinetic of hemolysis was measured spectrophotometrically at 700 nm. The kinetic 96-well microassay format was used for this purpose. We determined the t ½values (time required to lyse 50 % of target erythrocytes), which were employed to calculate the percentage of inhibition of the hemolytic activity promoted by each sample concentration. To confirm complement system activation, complement-dependent human neutrophil migration was examined using the Boyden chamber model.Results At the highest concentration tested (120 μg/mL), B. jararacussu and B. pirajai crude venoms inhibited the hemolytic activity of the classical pathway (65.3 % and 72.4 %, respectively) more strongly than they suppressed the hemolytic activity of the alternative pathway (14.2 and 13.6 %, respectively). BjussuSP-I (20 μg/mL) did not affect the hemolytic activity of the classical pathway, but slightly decreased the hemolytic activity of the alternative pathway (13.4 %). BpirLAAO-I (50 μg/mL) inhibited 24.3 and 12.4 % of the hemolytic activity of the classical and alternative pathways, respectively. Normal human serum treated with B. jararacussu and B. pirajai crude venoms induced human neutrophil migration at a level similar to that induced by zymosan-activated normal human serum.Conclusion Together, the results of the kinetics of hemolysis and the neutrophil chemotaxis assay suggest that pre-activation of the complement system byB. jararacussu and B. pirajai crude venoms consumes complement components and generates the chemotactic factors C3a and C5a. The kinetic microassay described herein is useful to assess the effect of venoms and toxins on the hemolytic activity of the complement system.(AU)

Immunomodulatory activity of Tityus serrulatus scorpion venom on human T lymphocytes

Background Tityus serrulatus scorpion venom (TsV) contains toxins that act on K + and Na + channels and account for the venom's toxic effects. TsV can activate murine peritoneal macrophages, but its effects on human lymphocytes have been poorly investigated. Considering that lymphocytes may play an important role in envenomation, we assessed whether TsV affects the expression of phenotypic (CD3, CD4, and CD8) and activation (CD69, CD25, and HLA-DR) markers, cell proliferation, and cytokine production in peripheral blood mononuclear cells. Methods Cytotoxicity of TsV was evaluated via the MTT assay. Cell proliferation, expression of phenotypic and activation markers, and release of cytokines were assessed using flow cytometry, after treatment with non-cytotoxic concentrations of TsV. The combined use of carboxyfluorescein diacetate succinimidyl ester and monoclonal antibodies against phenotypic and activation markers enabled us to simultaneously assess cell proliferation extent and cell activation status, and to discriminate among cell subpopulations. Results TsV at concentrations of 25 to 100 μg/mL were not cytotoxic towards peripheral blood mononuclear cells. TsV did not induce significant changes in lymphocyte subpopulations or in the expression of activation markers on CD4 + and CD8 + T cells. TsV inhibited the phytohemagglutinin-stimulated lymphocyte proliferation, particularly in the CD8 + CD25 + T lymphocyte subset. TsV alone, at 50 and 100 μg/mL, did not induce peripheral blood mononuclear cell proliferation, but elicited the production and release of IL-6, a proinflammatory cytokine that plays an important role in innate and adaptive immune responses. Conclusions TsV is a potential source of molecules with immunomodulatory action on human T lymphocytes.(AU)

Bothrops snake venoms and their isolated toxins, an L-amino acid oxidase and a serine protease, modulate human complement system pathways

Background Activation of the complement system plays an important role in the regulation of immune and inflammatory reactions, and contributes to inflammatory responses triggered by envenomation provoked byBothrops snakes. The present study aimed to assess whether Bothrops jararacussuand Bothrops pirajai crude venoms and their isolated toxins, namely serine protease (BjussuSP-I) and L-amino acid oxidase (BpirLAAO-I), modulate human complement system pathways.Methods Lyophilized venom and toxin samples solubilized in phosphate buffered saline were diluted in appropriate buffers to evaluate their hemolytic activity on the alternative and classical pathways of the complement system. Venom- and toxin-treated normal human serum was added to the erythrocyte suspension, and the kinetic of hemolysis was measured spectrophotometrically at 700 nm. The kinetic 96-well microassay format was used for this purpose. We determined the t ½values (time required to lyse 50 % of target erythrocytes), which were employed to calculate the percentage of inhibition of the hemolytic activity promoted by each sample concentration. To confirm complement system activation, complement-dependent human neutrophil migration was examined using the Boyden chamber model.Results At the highest concentration tested (120 g/mL), B. jararacussu and B. pirajai crude venoms inhibited the hemolytic activity of the classical pathway (65.3 % and 72.4 %, respectively) more strongly than they suppressed the hemolytic activity of the alternative pathway (14.2 and 13.6 %, respectively). BjussuSP-I (20 g/mL) did not affect the hemolytic activity of the classical pathway, but slightly decreased the hemolytic activity of the alternative pathway (13.4 %). BpirLAAO-I (50 g/mL) inhibited 24.3 and 12.4 % of the hemolytic activity of the classical and alternative pathways, respectively. Normal human serum treated with B. jararacussu and B. pirajai crude venoms induced human neutrophil migration at a level similar to that induced by zymosan-activated normal human serum.Conclusion Together, the results of the kinetics of hemolysis and the neutrophil chemotaxis assay suggest that pre-activation of the complement system byB. jararacussu and B. pirajai crude venoms consumes complement components and generates the chemotactic factors C3a and C5a. The kinetic microassay described herein is useful to assess the effect of venoms and toxins on the hemolytic activity of the complement system.

Immunomodulatory activity of Tityus serrulatus scorpion venom on human T lymphocytes

Background Tityus serrulatus scorpion venom (TsV) contains toxins that act on K + and Na + channels and account for the venoms toxic effects. TsV can activate murine peritoneal macrophages, but its effects on human lymphocytes have been poorly investigated. Considering that lymphocytes may play an important role in envenomation, we assessed whether TsV affects the expression of phenotypic (CD3, CD4, and CD8) and activation (CD69, CD25, and HLA-DR) markers, cell proliferation, and cytokine production in peripheral blood mononuclear cells. Methods Cytotoxicity of TsV was evaluated via the MTT assay. Cell proliferation, expression of phenotypic and activation markers, and release of cytokines were assessed using flow cytometry, after treatment with non-cytotoxic concentrations of TsV. The combined use of carboxyfluorescein diacetate succinimidyl ester and monoclonal antibodies against phenotypic and activation markers enabled us to simultaneously assess cell proliferation extent and cell activation status, and to discriminate among cell subpopulations. Results TsV at concentrations of 25 to 100 g/mL were not cytotoxic towards peripheral blood mononuclear cells. TsV did not induce significant changes in lymphocyte subpopulations or in the expression of activation markers on CD4 + and CD8 + T cells. TsV inhibited the phytohemagglutinin-stimulated lymphocyte proliferation, particularly in the CD8 + CD25 + T lymphocyte subset. TsV alone, at 50 and 100 g/mL, did not induce peripheral blood mononuclear cell proliferation, but elicited the production and release of IL-6, a proinflammatory cytokine that plays an important role in innate and adaptive immune responses. Conclusions TsV is a potential source of molecules with immunomodulatory action on human T lymphocytes.

Snake venom L-amino acid oxidases: an overview on their antitumor effects

The L-amino acid oxidases (LAAOs) constitute a major component of snake venoms and have been widely studied due to their widespread presence and various effects, such as apoptosis induction, cytotoxicity, induction and/or inhibition of platelet aggregation, hemorrhage, hemolysis, edema, as well as antimicrobial, antiparasitic and anti-HIV activities. The isolated and characterized snake venom LAAOs have become important research targets due to their potential biotechnological applications in pursuit for new drugs of interest in the scientific and medical fields. The current study discusses the antitumor effects of snake venom LAAOs described in the literature to date, highlighting the mechanisms of apoptosis induction proposed for this class of proteins.

Snake venom L-amino acid oxidases: an overview on their antitumor effects

The L-amino acid oxidases (LAAOs) constitute a major component of snake venoms and have been widely studied due to their widespread presence and various effects, such as apoptosis induction, cytotoxicity, induction and/or inhibition of platelet aggregation, hemorrhage, hemolysis, edema, as well as antimicrobial, antiparasitic and anti-HIV activities. The isolated and characterized snake venom LAAOs have become important research targets due to their potential biotechnological applications in pursuit for new drugs of interest in the scientific and medical fields. The current study discusses the antitumor effects of snake venom LAAOs described in the literature to date, highlighting the mechanisms of apoptosis induction proposed for this class of proteins.

Desregulação da apoptose em neoplasias mieloproliferativas crônicas / Apoptosis deregulation in myeloproliferative neoplasms

As neoplasias mieloproliferativas crônicas cromossomo Filadélfia negativas são doenças hematológicas clonais que se caracterizam pela independência ou pela hipersensibilidade dos progenitores hematopoiéticos às citocinas. Os mecanismos celulares e moleculares envolvidos na fisiopatologia das neoplasias mieloproliferativas crônicas ainda não estão totalmente esclarecidos. Achados fisiopatológicos relevantes para as neoplasias mieloproliferativas crônicas estão associados às alterações genéticas como, por exemplo, a mutação somática no gene que codifica o JAK2 (JAK2V617F). A desregulação do processo de morte celular programada, denominada apoptose, parece participar da patogênese dessas desordens. Sabe-se que a desregulação da expressão dos genes pró- e antiapoptóticos promove a resistência das células à apoptose, culminando com o acúmulo das células mieloides e estabelecendo a neoplasia. Esta revisão enfocou as alterações na regulação da apoptose em neoplasias mieloproliferativas crônicas e a importância da melhor compreensão desse mecanismo para o desenvolvimento de novas terapias para essas doenças.

Philadelphia-chromosome negative chronic myeloproliferative neoplasms are clonal hematologic diseases characterized by hematopoietic progenitor independence from or hypersensitivity to cytokines. The cellular and molecular mechanisms involved in the pathophysiology of myeloproliferative neoplasms have not yet been fully clarified. Pathophysiologic findings relevant for myeloproliferative neoplasms are associated with genetic alterations, such as, somatic mutation in the gene that codifies JAK-2 (JAK V617F). Deregulation of the process of programmed cellular death, called apoptosis, seems to participate in the pathogenesis of these disorders. It is known that expression deregulation of pro- and anti-apoptotic genes promotes cell resistance to apoptosis, culminating with the accumulation of myeloid cells and establishing neoplasms. This review will focus on the alterations in apoptosis regulation in myeloproliferative neoplasms, and the importance of a better understanding of this mechanism for the development of new therapies for these diseases.

Upregulation of hsa-miR-125b in HTLV-1 asymptomatic carriers and HTLV-1-associated myelopathy/tropical spastic paraparesis patients

The retrovirus human T lymphotropic virus type 1 (HTLV-1) promotes spastic paraparesis, adult T cell leukaemia and other diseases. Recently, some human microRNAs (miRNAs) have been described as important factors in host-virus interactions. This study compared miRNA expression in control individuals, asymptomatic HTLV-1 carriers and HTLV-1 associated myelopathy (HAM)/tropical spastic paraparesis patients. The proviral load and Tax protein expression were measured in order to characterize the patients. hsa-miR-125b expression was significantly higher in patients than in controls (p = 0.0285) or in the HAM group (p = 0.0312). Therefore, our findings suggest that miR-125b expression can be used to elucidate the mechanisms of viral replication and pathogenic processes.

Deregulated expression of A1, Bcl-2, Bcl-xL, and Mcl-1 antiapoptotic proteins and Bid, Bad, and Bax proapoptotic genes in polycythemia vera patients

Apoptosis deregulation might have a role in the pathophysiology of polycythemia vera (PV). This study evaluated Bcl-2 molecule expression in CD34+ cells and leukocytes in 12 PV patients. Gene expression was investigated by real time PCR using SybrGreen Quantitect kit and protein expression was evaluated by western-blotting. JAK2 V617F mutation was detected according to Baxter et al (2005). CD34+ cells from PV patients presented higher levels of A1 and Mcl-1 expression (median: 22.6 and 5.2, respectively) in comparison with controls (0.9 and 0.5, p=0.004 and p=0.020); while Bcl-2 and Bcl-xL expression decreased in PV patients (0.18 and 1.19) compared with controls (1.39 and 2.01, p=0.006 and p=0.020). CD34+ cells in PV patients showed an elevated Bid expression (14.4) in comparison with healthy subjects (1.0; p=0.002). Patients' leukocytes showed an A1 augmentation (7.41, p=0.001) and a reduced expression of Bax (0.19; p=0.040) and Bad (0.2; p=0.030). There was no correlation between JAK2 V617F allele burden and molecular expression. PV patients showed alterations in Bcl-2 members' expression, which may interfere with control of apoptotic machinery and contribute to disease pathogenesis.

A desregulação da apoptose parece participar da fisiopatologia da policitemia vera (PV). Este estudo avaliou a expressão das moléculas da família Bcl-2 em células hematopoéticas CD34 + e leucócitos de 12 pacientes com PV. Foram realizados: a quantificação da expressão gênica por PCR em tempo real utilizando kit Sybrgreen Quantitect, avaliação da expressão de proteínas por western-blot e detecção da mutação JAK2 V617F segundo Baxter et al. (2005). Células CD34 + dos pacientes com PV apresentaram maior expressão de A1 e Mcl-1 (mediana: 22,6 e 5,2, respectivamente) em comparação com controles (0,9 e 0,5, p = 0,004 e p = 0,020) e expressão de Bcl-2 e Bcl-xL diminuída nestes pacientes (0,18 e 1,19) em relação aos controles (1,39 e 2,01, p = 0,006 e p = 0,020). Células CD34 + dos pacientes com PV mostraram expressão elevada de bid (14,4) em comparação aos controles (1,0; p = 0,002). Leucócitos dos pacientes mostraram aumento de A1 (7,41, p = 0,001) e expressão reduzida do Bax (0,19; p = 0,04) e Bad (0,2; p = 0,030). Não houve correlação entre percentagem de alelos JAK2 V617F mutados e expressão molecular. Pacientes com PV apresentaram alterações na expressão de moléculas Bcl-2 que podem interferir no controle da apoptose e contribuir para a patogênese da doença.

Perfil microbiológico e desfechos clínicos de úlceras em pés de diabéticos internados / Microbiological profile and clinical outcome of severe foot ulcers of diabetic inpatients

Descrever o perfil microbiológico e os desfechos clínicos de úlceras graves em pés diabéticos de pacientes internados em um hospital universitário de atenção terciária no estado do Ceara, Brasil. Métodos: Conduziu-se uma análise retrospectiva de dados obtidos nos prontuários médicos de todos os pacientes diabéticos internados entre janeiro de 2006 a junho de 2007, nas enfermarias do Serviço de Endocrinologia e Diabetes do Hospital Universitário Walter Cantídio (Universidade Federal do Ceará), por úlceras graves em pés diabéticos, com no mínimo grau 2 da classificação de Wagner, refratárias ao tratamento ambulatorial. Dados clínicos (sexo, idade, tempo de diabetes e co-morbidades) de cada paciente assim como as características microbiológicas do material colhido das suas ulceras em pés ou das suas peças cirúrgicas (amputações) foram obtidos. Resultados: Foram identificados no período 17 diabéticos, todos tipo 2, com idade de 58,11±10,8 anos e 12,4±8,4 anos de doença, 58,8% homens. Das úlceras, 41,1% eram grau 2, 35,2% grau 3, 11,7% grau 4 e 11,7% grau 5 de Wagner, 64,7% com menos de 3 meses de evolução. Realizaram limpeza cirúrgica 82,3% dos pacientes e amputações 47%, sendo identificada osteomielite em 47% dos casos. Antibioticoterapia empírica foi iniciada em todos os pacientes, sendo ciprofloxacina/metronidazol o esquema mais usado (76,5%). Houve cultura negativa em 12,5% das realizadas. Nas positivas, os germes mais freqüentes foram: S. aureus (57,1%); S. viridans (28,7%); P. aeruginosas (28,7%) e M. morganii (28,7%) A maioria (75%) dos S. aureus isolados eram meticilino-resistentes, mas sensíveis à vancomicina. Conclusão: Observouse a presença de flora polimicrobiana com grande número de patógenos multirresistentes e elevada prevalência de osteomielite e amputações em diabeticos portadores de úlceras graves, neuropatia e doença vascular periférica.

To describe the microbiological profile and clinical outcomes of diabetic foot ulcers of inpatients of a tertiary university hospital, at Ceara, Brazil. Methods: We conducted a retrospective analysis of medical charts data of all diabetic inpatients of the Endocrine and Diabetes Unit of Walter Cantídio University Hospital (Federal University of Ceará), admitted from January, 2006 to June, 2007 for severe foot ulcers (minimum of grade 2 of Wagner`s classification), which were refractory to ambulatory treatment. Clinical data from each patient were recorded (sex, age, diabetes duration, and comorbidities) as well as microbiological characteristics of foot ulcers and surgical (amputations) material. Results: We identified 17 diabetic patients, all type 2, aged 58.11 ± 10.8 years and 12.4 ± 8.4 years of disease, 58.8% male. Of ulcers, 41.1% were grade 2; 35.2% grade 3; 11.7% grade 4 and 11.7% grade 5 of Wagner; 64.7% with less than 3 months of evolution. Debridement was performed in 82.3% of patients and amputation in 47%; osteomyelitis was identified in 47% of cases. All patients started empiric antibiotic therapy, where ciprofloxacin/metronidazole was the most used scheme (76.5%). Cultures were negative in 12.5% of the patients. In the positive ones, the most prevalent bacterial pathogens detected in the culture materials were: S. aureus (57.1%); S. viridans (28.7%); P. aeruginosas (28.7%); M. morganii (28.7%). The majority (75%) of isolated S. aureus were methicillin-resistant, but were sensitive to vacomicin. Conclusion: We observed the presence of polymicrobial flora with a large number of multiresistant pathogens and high prevalence of osteomyelitis and amputations in diabetic patients with severe ulcers, neuropathy and peripheral vascular disease.

Comparação antropométrica e do perfil glicêmico de idosos diabéticos praticantes de atividade física regular e não praticantes / Anthropometric comparison and the glycemic profile in ederly diabetics practitioners and non-practitioners of regular physical activity

O objetivo do estudo foi comparar os parâmetros antropométricos e perfil glicêmico de idosos diabéticos tipo 2 praticantes de atividade física regular e não praticantes. Métodos: O estudo, do tipo transversal, envolveu 70 idosos com diabetes mellitus tipo 2, com idade entre 60 e 80 anos. A coleta de dados foi através de questionário estruturado abordando: a) características demográficas dos pacientes (idade e nível de escolaridade) e b) características do padrão de atividade física (frequência, duração (min) e tempo (meses) da prática de exercícios físicos). Foram realizadas medidas antropométricas e verificação do perfil glicêmico (glico-hemoglobina). Os dados foram analisados no programa STATA versão 9.0. Foram calculados a média e desvio padrão da média (DP) e realizado o teste de análise de variância (ANOVA). Para verificar a associação de variáveis qualitativas, utilizou-se o teste exato de Fisher e as associações entre as variáveis quantitativas foram estimadas através do coeficiente de correlação de Pearson, com uma confiança de 95%. O nível de significância foi de p<0,05. Resultados: Em relação ao nível de escolaridade, não houve influência na realização da prática de atividade física. Nos PAF observa-se uma diminuição do IMC e do perfil glicêmico, quando comparados aos NPAF. A caminhada foi a atividade física mais realizada pelos idosos (51,5%). A partir dos resultados obtidos na análise dos dados, foi possível mostrar os benefícios que a atividade física proporciona aos indivíduos no processo de envelhecimento e no bom controle glicêmico.