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Introduction: DLBCL presents clinical, biological, and prognostic heterogeneity. The composition of the tumor TME strongly contributes for its biological diversity, being able to predict treatment response and determine prognosis. DLBCL with TME enriched in CD8+T-lymphocytes and NK cells demonstrate increased OS compared to those with TME enriched in histiocytes. Data from patients with solid tumors have established associations between absolute quantification of lymphocyte subpopulations (LS) in peripheral blood (PB) and prognosis. Therefore, it is hypothesized that the cellular constituents of PB may mirror the TME, and their quantification may constitute an easy and fast tool to access biological biomarkers predictive of therapeutic response and prognosis in malignancies. This study aims to determine the absolute quantification of LS in the PB of DLBCL patients, seeking associations with outcomes and specific clinical phenotypes. Methods: This study involved 54 newly-diagnosed DLBCL patients. PB collection was performed before starting any therapy, for blood count and immunophenotyping on a BD Facs Calibur flow cytometer. End points included OS and EFS. Survival curves were constructed using the KM method. The Mann-Whitney test and the Kruskal-Wallis test were used to establish the relationship between the lymphocyte profile, clinical variables, therapeutic response and outcomes. The Contal O'Quingley test was used to determine cut-offs for each LS capable of discriminating outcomes. Univariate analysis was performed using the Cox method and a p-value ≤ 0.05 was considered statistically significant. Results: The median age at diagnosis was 54 years and 55.6% were female. Clinical stage III/IV was observed in 77.8%, 57.4% had bulky ≥ 7 cm, 63.0% had B-symptoms, and 53.7% presented IPI ≥ 3. The ORR was 80.8%. With a median follow-up of 21.3 months, the estimated 2-year OS and EFS were 73.3% and 64.7%. The median absolute values obtained for the PB lymphocyte subpopulations were: 1026 cells for CD3+ T-lymphocytes, 591 cells for CD4+ T-helper lymphocytes, 412 cells for CD8+ T-cytotoxic lymphocytes, 1.6 for CD4/CD8 ratio, 240 cells for T-reg lymphocytes, 112 cells for LG T-lymphocytes, 173 cells for NK cells, and 70 cells for B-lymphocytes. In univariate analysis, using a cut-off value = 367 CD8+T-cells/mm3 and 141 NK-cells/mm3, cytotoxic T-lymphodepletion was associated with decreased OS (p = 0.005) and EFS (p < 0.001), as well as NK-lymphodepletion was associated with decreased EFS (p = 0.033), but not with shortened OS (p = 0.240). DLBCL patients presenting higher NK-cell counts had lower rates of chemoresistance to the R-CHOP (p = 0.024), and those with higher NK-cell counts (p = 0.033) and higher CD8+T-lymphocyte counts (p = 0.039) had lower probability to develop relapse, progression or death, confirming the central role of these PB cellular constituents in anti-tumor immunity. We observed that B-cell lymphodepletion was associated with an adverse clinical phenotype, including higher frequency of bulky disease (p = 0.009), B-symptoms (p < 0.001) and advanced clinical stage (p = 0.033). Conclusion: In newly-diagnosed DLBCL, the lymphodepletion of CD8+T-cells and NK cells quantified in PB were associated with decreased OS and EFS. Similarly, peripheral T-cytotoxic and NK lymphodepletion were associated with absence of response to the R-CHOP regimen, as well as higher probability of relapse, progression or death. Therefore, the absolute quantification of these LS in PB may be considered potential biomarkers capable of predicting response and prognosis in DLBCL. Also, we believe it may mirror the cellular composition of the immune TME.

Introduction: DLBCL commonly affects the older population. Elderly patients often present high-risk molecular profile, lower tolerability to chemotherapy, and poor outcomes. In this setting, attenuated therapeutic regimens, such as R-miniCHOP and R-miniCHOP of the elderly, have emerged for this fragile population. Albeit its relevance, response rates, clinical outcomes, and toxicities of these regimens remain poorly understood, because these individuals are usually excluded from controlled trials. Therefore, this study aims to assess the outcomes, determine survival predictors, and compare responses and toxicities between R-CHOP, R-miniCHOP, and R-miniCHOP of the elderly protocols in the management of elderly DLBCL patients. Methods: This retrospective, observational, and single-center study involved 185 DLBCL patients older than 70 years, treated at USP, from 2009 to 2020. Endpoints included ORR, OS, PFS. Survival curves were constructed using the KM method and the Log-Rank test was used to assess the relationship between variables and outcomes. The chi-square test and the Kruskal-Wallis test were applied to assess statistically significant differences in clinical characteristics, adverse event profile, and responses between different treatment modalities. Univariate analysis was performed using the Cox test and multivariate analysis by Cox regression method. The results were presented in HR and 95% CI, and a p-value ≤0.05 was considered significant. Results: The median age at diagnosis was 75 years, and 58.9% were female. Comorbidities were prevalent, including 19.5% with immobility, 28.1% with malnutrition and 24.8% with polypharmacy. Advanced clinical stage (III/IV) was observed in 72.4%, 48.6% had bulky ≥ 7 cm, 63.2% had B-symptoms, and 67.0% presented IPI ≥ 3. Among the 182 (98.4%) effectively treated cases, 57.1% received R-CHOP, 18.0% R-miniCHOP, 13.2% R-miniCHOP of the elderly, and 1.7% were palliated using R-CVP. The ORR for the whole cohort was 68.1%, with CR achieved in 65.9%. ORR was 72.1% for R-CHOP, 70.6% for R-miniCHOP, and 45.6% for R-miniCHOP of the elderly, p = 0.040. Although R-miniCHOP of the elderly regimen promoted lower ORR, patients in this group had higher rates of unfavorable clinical-laboratory findings, including hypoalbuminemia (p = 0.001), IPI ≥ 3 (p = 0.013), and NCCN-IPI ≥ 3 (p = 0.002). With a median follow-up of 6.3 years, the estimated 5-year OS and PFS were 50.2% and 44.6%, respectively. The estimated 2-year OS was 82.0% for R-CHOP, 67.5% for R-miniCHOP, and 48.1% for R-miniCHOP of the elderly, p = 0.003. The estimated 2-year PFS was 61.1% for R-CHOP, 56.4% for R-miniCHOP, and 20.5% for R-miniCHOP of the elderly, p = 0.005. Although correlated with increased OS and PFS in comparison to attenuated protocols, R-CHOP regimen was associated with higher rates of severe neutropenia (p = 0.003), but not translated into febrile neutropenia (p = 0.907), therapy interruption (p = 0.671), or higher early mortality rates (p = 0.681). In multivariate analysis, age≥75 years (HR: 2.08, p = 0.001), neutrophilia (HR: 2.18, p = 0.007), low lymphocyte/monocyte ratio (HR: 1.98, p = 0.010), and clinical stage III/IV (HR: 2.36, p = 0.003) were predictors of decreased OS. Conclusion: In this large and real-life LATAM cohort, we demonstrated that DLBCL patients older than 70 years still do not have satisfactory outcomes, with half of cases not reaching 5 years of life expectancy after diagnosis. Although a significant portion of older DLBCL patients is highly fragile and ineligible for enhanced regimens, attenuated anthracycle-based protocols promoted remarkably inferior outcomes compared to those achieved by the R-CHOP.

Introduction: FL has heterogeneous biological behavior, variable clinical outcomes, and a natural history marked by histological transformation (HT). The management of FL and its outcomes are highly dependent on staging (CS) and tumor burden (TB). The identification of new molecular biomarkers is crucial, not only to understand FL's biology, but also, can guide better prognostic stratification, and determine new therapeutic targets. This study aims to outline the GEP in tissue samples from FL categorized by CS and TB using a large-scale gene microarray platform with subsequent expansion by RT-qPCR. Methods: This study involved 219 FL cases. FL patients were categorized into early-stage disease (I/II) (ES), advanced-stage (AS) (III/IV) with low tumor burden (LTB), and AS with high tumor burden (HTB) according to GELF criteria. The experimental phase involved 126 FFPE samples. Firstly, 42 samples (12 ES, 11 AS-LTV, 12 AS-HTV, 6 control lymph nodes and 1 control HELA) were submitted to the array using the Clariom D Array Human platform. The identification of differentially expressed genes (DEGs) between the 4 groups used the Transcriptome Analysis Console by the eBay ANOVA method. Bidimensional PCA sought to identify discriminatory DEGs clusters. A comparison between pairs was subsequently performed. The 7 most discriminatory DEGs were selected for a cohort expansion by RT-qPCR. Results: According to CS and TB, 14.6% of FL cases were categorized as ES, 17.8% as AS-LTB, and 67.6% as AS-HTB. With a median follow-up of 80.7 months, the median OS was 208.2, 130.5, and 126.3 months for ES, AS-LTB, and AS-HTB,p = 0.249. The median PFS was 113.3, 110.7, and 59.5 months, for the same groups, p = 0.018. The array identified 135,750 genes comparisons between the control, ES, AS-LTB, and AS-HTB. PCA analysis was not able to identify discriminatory DEGs clusters among the 4 groups. Pairwise comparative analysis identified 1913 DEGs among the 4 groups. Of these, 1759 (91.9%) were DEGs between FL and healthy controls, but only 218 (11.3%) were DEGs among the 3 different FL subgroups, evidencing a low molecular diversity in FL when categorized by CS and TB. In the cohort expansion by qRT-PCR, 7 pre-selected DEGs based on the microarray were tested, including the genes GNG7, ITGAM, AZN1, TIMD4, IL7R, PTGS2, and MTTL10. The comparison between the median expression of these genes among the 3 FL's groups showed that the expression of the ILR7 was downregulated with the progression of the disease, with marked hypoexpression in AS-HTB patients. These data may suggest downregulation of IL7R gene in the FLs natural history. We found that hypoexpression of the GNG7 (p = 0.024) and PTGS2 (p = 0.090) were associated with higher rates of disease progression/relapse, and hypoexpression of the GNG7 (p = 0.006) was associated with a higher probability of HT, highlighting the potential prognostic impact of these molecular biomarkers. Finally, we established phenotypic-genotypic associations, as hypoexpression of ITGAM was associated with higher rates of B-symptoms (p = 0.017) and bulky disease (p = 0.026), as well as hypoexpression of IL7R was correlated with a higher probability of bulky (p = 0.025) and extranodal involvement (p = 0.016). Conclusion: Although CS and TB are strong determinants of the prognostic heterogeneity of FL, this study did not find high molecular diversity by this categorization. We observed a relatively homogeneous GEP with low biological diversity in FL presenting different disease evolution phases. Moreover, new molecular biomarkers potentially implicated in FLs natural history, prognostication and specific clinical phenotypes were revealed.

Objetivos: Avaliar a frequência da utilização de profilaxia de tromboembolismo venoso (TEV) em uma coorte de pacientes clínicos através da aplicação de ferramentas de acesso de risco trombótico e risco hemorrágico. Material e métodos: Realizada análise observacional, longitudinal e retrospectiva dos dados de prontuários de pacientes internados nas enfermarias de Clínica Médica (CM) de Hospital de Ensino da Região Metropolitana da Baixada Santista, de janeiro a março de 2022. Os critérios de elegibilidade foram idade ≥ 18 anos e internação em enfermaria de CM com período mínimo de 24 horas após admissão hospitalar. Foram excluídos da análise pacientes com diagnóstico de COVID-19 e pacientes submetidos a cirurgia. Para adequada avaliação do risco trombótico, aplicamos o Escore de Pádua utilizando dados recuperados de prontuários e, para avaliação do risco hemorrágico, utilizamos o Escore IMPROVE. Foi considerada tromboprofilaxia adequada, o uso de HBPM ou heparina não fracionada, esta especialmente nos pacientes com contraindicação a HBPM, ex, pacientes com clearence de creatinina inferior a 30ml/min. Pacientes que não receberam tromboprofilaxia medicamentosa por contraindicação formal como plaquetometria inferior a 50.000/μL, foram categorizados como adequados do ponto de vista de manejo de profilaxia trombótica. Não foram avaliadas medidas mecânicas de tromboprofilaxia e não observamos alertas eletrônicos em prontuário para otimizar profilaxia de TEV. Resultados: Após análise consecutiva de 191 prontuários, 157 preencheram critérios de inclusão. O tempo médio de internação foi de 22 dias, a maioria dos pacientes eram do sexo feminino (55.7%), com idade entre 41-70 anos (66.5%), 4.4% dos casos apresentava idade > 85 anos. De comorbidades mais prevalentes, 65% tinham câncer em atividade (15.9% hematológica) e 40.1% insuficiência cardíaca ou respiratória. No IMC, 21% tinham sobrepeso e 11.4% obesidade. Cabe ressaltar que 7.6% apresentou hemorragia 3 meses prévios à admissão. Segundo Pádua, 71.9% foram alocados em alto risco trombótico e, dentre esses, 47,6% receberam profilaxia adequada. Quanto ao risco hemorrágico, 28.6% foram alocados como alto risco. Destes, 55,5% foram submetidos a profilaxia medicamentosa. Discussão: Observamos que a maioria da coorte tinha indicação para profilaxia medicamentosa e quase metade não a recebeu. Apesar de se tratar de enfermaria não setorizada, não oncológica, grande parte dos pacientes tinham câncer, agregando maior risco trombótico, dificultando profilaxia, visto temor de evento hemorrágico nestes casos. Não observamos em dados de prontuário referência a indicação ou contraindicação a profilaxia de TEV. Não existe no serviço ferramenta eletrônica de alerta à equipe médica para melhoria do uso de profilaxia de TEV. Os resultados obtidos neste estudo estão de acordo com outros já publicados. Um estudo publicado na Associação Médica Brasileira em 2013, realizado em uma enfermaria de CM na cidade de Marília, evidenciou que quase metade dos 146 pacientes analisados durante internação não receberam correta tromboprofilaxia. Conclusão: Observamos subutilização de profilaxia de TEV, enxergamos potencial de melhora com criação/divulgação de protocolo institucional e utilização de ferramentas eletrônicas na tentativa de diminuição da morbimortalidade relacionada ao TEV.

Objetivos: Relatar caso de disceratose congênita (DC), diagnóstico tardio, na 4ªdécada de vida, apesar da presença de dados de história e apresentação clínica clássica que compõem a doença estarem presentes no paciente. Material e métodos: Paciente masculino, 44 anos, referenciado para hemato por pancitopenia grave. Referia astenia crônica, dispneia progressiva há 4 meses e febre. Ao EF estava descorado, com anoníquia, máculas hipocrômicas disseminadas e leucoplasia oral. Referia alterações de pele e fâneros há anos, sem precisar quantos. Os antecedentes pessoais se limitavam a contraindicação a procedimentos cirúrgicos por problemas de coagulação, SIC, negando outras comorbidades. Já tinha recebido hemocomponentes em internações pregressas em outros serviços de emergência. De história relevante, tinha um irmão com óbito secundário a câncer de esôfago, e outro irmão com câncer de reto, em quimioterapia. Questionado, confirmou a presença de alterações de pele e fâneros nos dois irmãos. De exames, o hemograma de admissão demonstrava pancitopenia (Hb de 2,5 g/dL, Leuco 3600/mm3 neutrófilos 880 plaquetas e 3.000/mm3). Feito mielograma, com hiperplasia eritróide, ninhos de eritroblastos, pontes intercitoplasmáticas, figuras de mitose, blebs citoplasmáticos. Analise imunofenotípica sem fenótipo anômalo. BMO com 20% de celularidade. Citogenética normal (46,XY[20]). Frente a suspeita de falência medular hereditária, feito painel NGS customizado com pesquisa dos genes ACD, CTC1, DKC1, NAF1, NHP2, NOP10, PARN, RTEL1, STN1, TERC, TERT, TINF2, TYMS, WRAP53, com resultado positivo de variante patogênica para DKC1, confirmando DC. Devido à vulnerabilidade social do paciente, dificuldades de ida a um centro transplantador, foi iniciado terapia androgênica, mas o paciente não retornou para acompanhamento, presumindo-se um desfecho desfavorável para o caso. Discussão: A DC é uma condição genética rara, marcada por instabilidade cromossômica, cuja fisiopatologia envolve mutações nos genes de regulação telomérica, gerando encurtamento precoce destes segmentos e envelhecimento celular prematuro em tecidos que se renovam rapidamente. Existe um alto risco de desenvolvimento de neoplasias malignas, especialmente na vida adulta. A tríade clássica mucocutânea de distrofia ungueal, leucoplasia oral e pigmentação reticulada cutânea estão presentes em cerca de 50% dos pacientes, o que pode guiar o diagnóstico clinico ou mesmo precoce da doença, já que quase 75% dos casos tem ao menos um destes achados clínicos. De difícil acesso no sistema público, a análise do comprimento do telômero com complementação de painel genético confirmam o diagnóstico. A principal causa de morte é a falência medular, com anemia grave, aumento do risco de sangramento e infecções, além de neoplasias. O único tratamento curativo é o transplante de medula óssea, normalmente em centros especializados em falências medulares hereditárias, ainda mais difíceis de acesso na rede pública. Conclusão: Este caso destaca a importância da necessidade do diagnóstico precoce para tentativa de melhores desfechos. Chama a atenção a riqueza de achados de exame físico incomuns, como anoníquia, leucoplasia oral e pigmentação reticulada cutânea e a história familiar de malignidades. Apesar de ser uma doença rara, o paciente apresentava a tríade clássica da DC, associado a falência medular grave, e confirmação molecular.

Objetivo: Relatar caso raro de anemia aplástica no contexto gestacional, abordagem terapêutica, complicações e desfechos. Material e métodos: Paciente feminino, 19 anos, gestante com idade gestacional de 16 semanas, foi encaminhada pelo obstetra para internação na hematologia em 16 de fevereiro de 2024 devido a queixa de fraqueza, astenia, cefaleia e hemograma com bicitopenia (anemia e plaquetopenia). Ao EF estava descorada, sem petéquias, e com propedêutica obstétrica normal para idade gestacional. Interrogada, negava sangramentos ativos, mas referia episódios isolados de gengivorragia. De AP, a mãe referia 01 episódio de hemotransfusão sem precisar diagnóstico, e uso de ácido fólico e sulfato ferroso. Laboratorialmente, apresentava Hb de 7.4 g/dl, VCM 94.6, leucócitos 6030, neutrófilos 4221 e plaquetas 34.000/mm3. Apresentava reticulocitopenia e provas de hemólise negativas. A dosagem de Ferritina foi de 1.050 ng/ml, saturação 35%, vitamina B12 de 219 pg/ml (221-911 pg/ml), optado então por reposição IM. Todas as sorologias foram negativas. Após reposição, houve incremento de 01 g/dl na Hb, e de plaqueta para 69.000 ainda durante internação, com HD de anemia carencial e seguimento ambulatorial. Um mês após reposição de B12 em doses adequadas, a paciente retorna com nova queda da Hb e plaquetometria, com necessidade transfusional de dois concentrados de hemácias. Revisitado diagnóstico com avaliação medular com mielograma normocelular, cariótipo 46,XX[20], imunofenotipagem sem fenótipo anômalo, BMO com de 30% de células hematopoiéticas, imunohistoquímica com celularidade de 25 a 30%, ausência de fibrose, e clone HPN negativa. Feito diagnóstico de anemia aplástica não grave, e indicado tratamento com ciclosporina monoterapia. Com melhora da citopenia e independência transfusional, a paciente recebeu alta com Hb de 7,9 g/dl e plaquetas de 40.000/mm3 para seguimento ambulatorial conjunto com hematologia e gestação de alto risco. De intercorrências do tratamento, a paciente apresentou hipertrofia gengival e, em julho de 2024, os últimos exames demonstraram melhora significativa dos parâmetros laboratoriais: HB 9,3 g/dl e plaquetas 165.000/mm3 e a paciente segue em acompanhamento a cada 15 dias para controle, com 37 semanas de gestação, com proposta de indução com 39 semanas pela equipe de obstetrícia. Discussão: A anemia aplástica (AA) é um distúrbio raro caracterizado por pancitopenia e hipocelularidade em MO que, associada à gestação, pode levar a complicações materno-fetais graves como hemorragia, sepse, pré-eclâmpsia, crescimento fetal inadequado e parto prematuro. Chen e cols demonstraram em coorte retrospectiva (n = 18) que nenhuma gestante apresentava neutropenia ≤ 500, com 83.3% da amostra não neutropênica. A ausência de gatilho infeccioso, displasia citomorfológica, cariótipo normal, toxicidade medicamentosa, e a resposta à imunossupressão apontam para aplasia que, no contexto da gestação, tem fisiopatologia desconhecida. É incerto se alterações hormonais ou imunes da gestação tenham relação etiopatogênica. Outro desafio envolve qual terapêutica é ideal, já que tratamentos convencionais, como TMO, não estão indicados. Neste contexto, a ciclosporina é opção relativamente segura para o concepto. Conclusão: Citopenias na gestante, embora frequentes, devem ser adequadamente avaliadas pela equipe obstétrica com suporte do hematologista em razão da complexidade tanto diagnóstica, quanto terapêutica.

Objetivos: O estudo visa avaliar retrospectivamente os encaminhamentos da atenção primária de saúde (APS) para a hematologia, tentando identificar padrões de encaminhamento, avaliar a complexidade do diagnóstico, e propor melhorias no processo. Material e métodos: Estudo retrospectivo de dados de prontuário dos casos encaminhados da APS, fevereiro de 2023 a junho de 2024, para o ambulatorio de hematologia da Faculdade de Ciencias Medicas de Santos/UNILUS. Foram analisados dados epidemiológicos como idade, sexo, motivo do encaminhamento, natureza benigna ou maligna da doença suspeita e hipotese e/ou conclusão diagnostica. Resultados: Foram analisados 169 pacientes, 109 mulheres (64,50%) e 60 homens (35,50%), com idade mínima de 17 anos, máxima de 94 anos e média de 56,38 anos. Inicialmente, os pacientes foram divididos em 2 grupos: hematologia clássica (86,40%) e oncohematologia (13,60%). Todos os pacientes com neoplasia maligna foram alocados como doença de difícil manejo e necessidade de especialista. Na hematologia clássica, realocamos em doença de fácil manejo onde o diagnóstico e tratamento poderiam ser conduzidos na APS. O outro grupo, doenças de difícil manejo, o encaminhamento ao serviço de hematologia é justificável pois requer expertise/recursos não disponíveis na ASP. Após estratificação e analise, entendemos que 35,50% de nossa coorte eram casos de fácil manejo. Já as doenças de encaminhamento necessário ao especialista, representaram 64,50% da amostra. As doenças de mais fácil manejo foram representadas por anemias carenciais (deficiência de ferro sem anemia, anemia ferropriva, anemia megaloblástica, anemia multicarencial); anemias de doença crônica/inflamação (incluindo anemia da doença renal crônica e anemia da Insuficiência cardíaca), e traço falciforme. Observamos encaminhamentos por alteração laboratorial no hemograma como leucocitose reacional, linfopenia, pseudoplaquetopenia, e plaquetose cujo diagnóstico primário era bastante evidente. Como alteração de exame físico, recebemos um paciente purpura senil leve. As doenças de difícil manejo incluíam as anemias não alocadas acima, hemoglobinopatias (SS, SC, talassemia), neutropenia com infecção, coagulopatias, plaquetopenias sem causa aparente, SAF, trombofilia, PTI e poliglobulia. Embora tivéssemos alocado poliglobulia como difícil manejo, todos as poliglobulias tinham causa secundária aparente - tivemos grande dificuldade de acesso a exames como polissonografia e mutação do jak2 para retorno do paciente à APS com diagnóstico concluído. Discussão: A despeito da universalidade do SUS, o tempo de espera em consultas especializadas é longo. Desconhecemos fluxo definido na priorização do atendimento. Isto coloca todos os pacientes na mesma lista de espera. Observamos que parte significativa dos encaminhamentos são injustificáveis (35,5%). Preocupante, é a não otimização do acesso à recurso especializado, a sobrecarga do serviço, o tempo de espera no atendimento, e o preparo dos profissionais da APS para doenças de fácil manejo, como anemia carenciais, por exemplo. Felizmente, apenas 13,6% dos casos se tratava de doença oncohematológica. Conclusão: É necessário aprimorar o atendimento médico através da capacitação da equipe na APS, e otimizar os recursos através de fluxos de encaminhamento.

Introduction: Mantle cell lymphoma (MCL) is a rare malignancy with heterogeneous behavior. Despite the therapeutic advances recently achieved, MCL remains incurable. Currently, the standard of care for young and fit patients involves induction immunochemotherapy followed by up-front ASCT. However, the role of more intensive induction regimens, such as those based on high-doses of cytarabine (HDAC) remains controversial in the management of ASCT-eligible patients. Based on this premise, the current study aims to describe clinical and laboratory characteristics, assess outcomes, determine survival predictors, and compare responses between different primary therapeutic strategies, with focus on assessing the impact of HDAC-based regimens on outcomes in ASCT-eligible patients. Methods: This retrospective, observational, and single-center study involved 165 MCL patients, diagnosed and treated at the University of São Paulo, Brazil, from 2010 to 2022. Endpoints included overall OS, EFS, and POD-24 from up-front treatment. Survival curves were constructed using the Kaplan-Meier method and the Log-Rank test was used to assess the relationship between variables and outcomes. Univariate analysis was performed using the semi-parametric Cox test and multivariate analysis by Cox regression method or proportional ratios model. The results were presented in HR and 95% CI, and a p-value ≤ 0.05 was considered statistically significant. Results: The median age at diagnosis was 65 years and 73.9% were male. More than 90% of cases had a classic nodal variant (cnMCL), 76.4% had bone marrow infiltration, and 56.4% presented splenomegaly. Bulky ≥ 7 cm, B-symptoms, ECOG ≥ 2, and advanced-stage III/IV were observed in 32.7%, 64.8%, 32.1%, and 95.8%, respectively. Sixty-four percent of patients were categorized as high-risk according to MIPI score. With a median follow-up of 71.1 months (95% CI: 61.8-80.4) the estimated 2-year OS and EFS were 64.1% (95% CI: 56.2-71.9%) and 31.8% (95% CI: 23.0-40.6%), respectively. The ORR for the whole cohort was 64.7% (95% CI: 57.1-71.7%), with CR achieved in 41.5% (95% CI: 34.1-49.1%). POD-24 was observed in 46.7% (95% CI: 39.1-54.3%), and the overall mortality rate during all follow-up was 58.2% (95% CI: 50.6-65.5%), with disease progression being the main cause of death. Patients treated with R-HDAC-based regimens (43.4%-66/152) had higher ORR (85.9% vs 65.7%, p = 0.007) compared to those receiving R-CHOP (46.0%-70/152), as well as lower POD-24 rates (61.9% vs 80.4%, p = 0.043) and lower mortality (43.9% vs 68.6%, p = 0.004). However, intensified induction regimens using R-HDAC were not associated with a real OS benefit in MCL patients undergoing up-front consolidation with ASCT (2-year OS: 88.7% for R-HDAC plus ASCT vs 78.8% for R-CHOP plus ASCT, p = 0.289). Furthermore, up-front ASCT was independently associated with increased OS (p < 0.001), EFS (p = 0.005), and lower POD-24 rates (p < 0.001) in MCL. Additionally, CNS involvement (HR: 3.12, p = 0.004), tumor lysis syndrome (HR: 2.79, p = 0.026), albumin < 3.5 g/dL (HR: 2.69, p < 0.001), and failure to achieve remission after induction therapy (HR: 3.71, p < 0.001) were predictors of poor OS. Conclusion: In the largest Latin American cohort of MCL reported to date, we confirmed the OS benefit promoted by up-front consolidation with ASCT in young anf fit patients, regardless of the intensity of the immunochemotherapy regimen used in the pre-ASCT induction. Although HDAC-based regimens were not associated with an unequivocal increase in OS, it was associated with higher ORR and lower rates of early-relapses.

Introduction: FL is the most common low-grade B-cell NHL. Although indolent, it has heterogeneous biological behavior and variable clinical outcomes. Histologic transformation (HT) to high-grade B-cell NHL is associated with dismal survival. The therapeutic management of FL and its prognosis are highly dependent on its staging and tumor burden. Here we aimed to describe clinical characteristics, assess outcomes, determine predictors of survival and HT, and compare responses between different therapies applied in a large cohort of FL patients. Methods: This retrospective study involved 223 patients with FL G1-3A, diagnosed at USP, Brazil, from 2006 to 2022. FL patients were categorized into early-stage (I/II) (ES), advanced-stage (AS) (III/IV) with low tumor burden (LTV), and AS with high tumor burden (HTV) according to the GELF criteria. Endpoints were OS, PFS, early-relapse (< 24 months) and HT rates. Survival curves were constructed using the KM method and the Log-Rank test was used to assess the relationship between variables and outcomes. Univariate and multivariate analysis were performed. The results were presented in HR and 95% CI and p ≤ 0.05 was considered significant. Results: The median age at diagnosis was 60 years and 54.8% were female. 15% of cases had ES, 18.4% had AS-LTV, and 66.8% had AS-HTV. BM involvement, leukemic presentation, splenomegaly, and serous effusions occurred in 48.9%, 11.7%, 12.6%, and 21.1%, respectively. Bulky ≥ 7 cm, B-symptoms, ECOG ≥ 2, and involvement of ≥ 4 nodal areas were observed in 44.4%, 51.6%, 15.2%, and 56%, respectively. 49% of patients were categorized as high-risk according to the FLIPI score. With a median follow-up of 79.5 months (95% CI 67.0-92.0) the OS medians were 18 years (95% CI 14.9-21.1), 11 years (95% CI 9.2-12.9), and 10.5 years (95% CI 9.3-11.8) for ES, AS-LTV, and AS-HTV, respectively, p = 0.138. Similarly, the PFS medians were 6.7 years (95% CI 3.6-9.8), 8.4 years (95% CI 2.0-14.8), and 3.5 years (95 CI 1.8-5.3) for ES, AS-LTV, and AS-HTV, respectively, p = 0.171. Early-relapses occurred in 36.3% of cases, being 18.2%, 31.7%, and 41.0% for ES, AS-LTV, and AS-HTV, respectively, p = 0.032. The overall mortality rate was 29.1% for the whole cohort. HT was documented in 16.7% of cases, being 3.0%, 14.6% and 20.1% for ES, AS-LTV, and AS-HTV, respectively, p = 0.053. Among patients with AS-HTV, the R-CHOP regimen did not promote increased OS compared to the R-CVP regimen (p = 0.415), however it was associated with a substantial increase in PFS (p = 0.005). Age ≥ 60 years (HR 1.06, p < 0.001), ≥ 2 comorbidities (HR 3.85, p = 0.014), B-symptoms (HR 2.35, p = 0.015), HT (HR 2.77, p = 0.002) and thrombocytopenia (HR 2.96, p = 0.028) were predictors of poor OS. Similarly, age ≥ 60 years (HR 1.08, p < 0.001), involvelment of ≥ 4 nodal areas (HR 1.27, p < 0.001), and high LDH (HR 1.65, p = 0.002) predicted decreased PFS. Serous effusions (HR 2.09, p = 0.05), albumin < 3.5 g/dL (HR 2.82, p = 0.05), B-symptoms (HR 2.00, p = 0.006), involvement of ≥ 4 nodal areas (HR 1.21, p = 0.014), and BM infiltration (HR 2.11, p = 0.05) were the main predictors for HT. Conclusion: Although it is characteristically an indolent disease, we demonstrated that a significant portion of FL patients have shortened survival. We confirmed this prognostic heterogeneity, particularly considering the clinical staging and tumor burden. Therefore, FL patients with AS-HTV had higher HT rates and early-relapses, both classic adverse prognostic markers, as well as a tendency to higher mortality. Additionally, we identified clinical and laboratory predictors for HT, which may in a near future direct adapted-risk therapeutic strategies.

Introduction: DLBCL is a malignancy with marked clinical, biological and prognostic heterogeneity. Its different outcomes may be predicted by the identification of molecular biomarkers. Therefore, quantification of circulating cell-free DNA (ccfDNA) and circulating tumor DNA (ctDNA) have emerged as minimally invasive methods able to predict its prognosis. However, such measurements are based on the use of costly technologies and with restricted access in clinical practice. Here, we aimed to quantify ccfDNA in DLBCL using capillary electrophoresis (CEF), a low-cost and widely available methodology, in order to establish prognostic associations between its serum concentrations, clinical characteristics and outcomes. Methods: This prospective study involved 54 DLBCL patients, diagnosed at USP, from 2019 to 2020, and 23 healthy-controls. All cases were treated with 6-8 cycles of R-CHOP. Plasma samples were collected from DLBCL at three moments: at diagnosis, after the 4th CT cycle and at the end of treatment. ccfDNA was extracted using the QIAamp MinElute ccfDNA kit and quantified by CEF (Agilent 2100 Bioanalyzer). Comparison between two groups was performed using the Mann-Whitney test, and between three groups using the Kruskall-Wallis test. The cut-off for [ccfDNA] (1147 ng/μl) was determined by ROC methodology. Survival curves were constructed using the KM method and Log-Rank test was used to assess the relationship between variables and outcomes, and p≤0.05 was considered significant. Results: The median age at diagnosis was 57 years and 61.1% were female. Advanced-stage was observed in 59.3%, 29.6% had ECOG ≥ 2, 52.6% had IPI ≥ 3, and 44.4% had bulky ≥ 10 cm. BM infiltration and involvement of ≥ 2 EN sites occurred in 5.6% and 16.8%, respectively. With a median follow-up of 32 months, the medians OS and EFS were 32 months (95% CI 28-35) and 31 months (95% CI 28-34), respectively. The ORR was 76% (95% CI 63.5-86). The overall mortality rate was 20.4% (95% CI 11.1-32.3). ccfDNA concentrations were significantly higher in DLBCL compared to controls (p < 0.001), however, it did not differ significantly between samples from DLBCL collected at the 3 different time points (p = 0.602). We observed positive associations between ccfDNA [ ] in diagnostic DLBCL samples and biomarkers of high tumor burden, including higher LDH levels (p = 0.003), bulky (p = 0.011), ECOG ≥ 2 (p < 0.001), and IPI ≥ 3 (p = 0.069). However, ccfDNA[ ] at diagnosis were not statistically associated with ORR (p = 0.264), nor with clinical outcomes, including OS (1-year OS: 92% for [ccfDNA] < 1147 ng/μl vs 76% for [ccfDNA] ≥ 1147 ng/μl, p = 0.217) and EFS (1-year EFS: 92% for [ccfDNA] < 1147 ng/μl vs 69% for [ccfDNA] ≥ 1147 ng/μl, p = 0.270). We believe that the short follow-up time and sample number restriction may have contributed to the lack of statistically significant association between ccfDNA [ ] and responses/outcomes observed. In this sense, cohort expansion and NGS techniques for ctDNA identification are ongoing to assess its potential impact as a prognostic biomarker in our cohort. Conclusion: We demonstrated that ccfDNA [ ] measured by CEF were able to accurately discriminate healthy individuals from DLBCL, highlighting its potential role as a minimally invasive diagnostic biomarker. Furthermore, ccfDNA [ ] at diagnosis in DLBCL patients were positively associated with markers of high tumor burden and adverse prognosis. Our results suggest that measurement of ccfDNA by CEF may be a highly cost-effective methodology to identify high-grade B-cell NHLs and recognize DLBCL subgroups with adverse clinical-prognostic characteristics.

Extranodal NK-/T-cell lymphoma (ENKTCL) is a rare and highly aggressive malignancy with significant racial and geographic variations worldwide. In addition to the formerly "nasal-type" initial description, these lymphomas are predominantly extranodal in origin and typically cause vascular damage and tissue destruction, and although not fully understood, Epstein-Barr virus (EBV) has an important role in its pathogenesis. Initial assessment must include a hematopathology review of representative and viable tumor areas without necrosis for adequate immunohistochemistry studies, including EBV-encoded small RNA (EBER) in situ hybridization (ISH). Positron emission tomography with 18-fluorodeoxyglucose ( 18 F-FDG-PET/CT) for accurate staging is essential, and most patients will have localized disease (IE/IIE) at diagnosis. Apart from other T-cell malignancies, the best treatment even for localized cases is combined modality therapy (chemotherapy plus radiotherapy) with non-anthracycline-based regimens. For advanced-stage disease, l-asparaginase-containing regimens have shown improved survival, but relapsed and refractory cases have very poor outcomes. Nowadays, even with a better understanding of pathogenic pathways, up-front therapy is completely based on chemotherapy and radiotherapy, and treatment-related mortality is not low. Future strategies targeting signaling pathways and immunotherapy are evolving, but we need to better identify those patients with dismal outcomes in a pre-emptive way. Given the rarity of the disease, international collaborations are urgently needed, and clinical trials are the way to change the future., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Costa, Pereira, Lage and Baiocchi.)

Background: Nodal peripheral T-cell lymphoma (nPTCL) constitute a heterogeneous group of neoplasms with aggressive behavior and poor-survival. They are more prevalent in Latin America and Asia, although data from Brazil are scarce. Its primary therapy is still controversial and ineffective. Therefore, we aim to describe clinical-epidemiological characteristics, outcomes, predictors factors for survival and compare the results of patients treated with CHOP and CHOEP regimens., Methods: Retrospective, observational and single-center study involving 124 nPTCL patients from Brazil treated from 2000 to 2019., Results: With a median follow-up of 23.7 months, the estimated 2-year overall survival (OS) and progression-free survival (PFS) were 59.2% and 37.3%, respectively. The median age was 48.5 years and 57.3% (71/124) were male, 81.5% (101/124) had B-symptoms, 88.7% (110/124) had advanced disease (stage III/IV) and 58.1% (72/124) presented International Prognostic Index (IPI) score ≥3, reflecting a real-life cohort. ORR to first-line therapy was 58.9%, 37.9% (N = 47) received CHOP-21 and 35.5% (N = 44) were treated with CHOEP-21; 30.1% (37/124) underwent to consolidation with involved field radiotherapy (IF-RT) and 32.3% (40/124) were consolidated with autologous hematopoietic stem cell transplantation (ASCT). The overall response rate (ORR) was similar for CHOP-21 (76.6%) and CHOEP-21 (65.9%), P = .259. Refractory disease was less frequent in the CHOEP-21 group (4.5% vs. 21.2%, P = .018). However, few patients were able to complete 6-cycles of CHOEP-21 (31.8%) than to CHOP-21 (61.7%), P = .003. Delays ≥2 weeks among the cycles of chemotherapy were more frequent for patients receiving CHOEP-21 (43.1% vs. 10.6%), P = .0004, as well as the toxicities, including G3-4 neutropenia (88% vs. 57%, P = .001), febrile neutropenia (70% vs. 38%, P = .003) and G3-4 thrombocytopenia (63% vs. 27%, P = .0007). The 2-year OS was higher for CHOP (78.7%) than CHOEP group (61.4%), P = .05, as well as 2-year PFS (69.7% vs. 25.0%, P < .0001). In multivariate analysis, high LDH (HR 3.38, P = .007) was associated with decreased OS. CR at first line (HR: 0.09, P < .001) and consolidation with ASCT (HR: 0.08, P = .015) were predictors of increased OS., Conclusion: In the largest cohort of nPTCL from Latin America, patients had poor survival and high rate of chemo-resistance. In our cohort, the addition of etoposide to the CHOP-21 backbone showed no survival benefit and was associated with high-toxicity and frequent treatment interruptions. Normal LDH values, obtaintion of CR and consolidation with ASCT were independent factors associated with better outcomes., Competing Interests: Disclosures The authors declare that they have no competing interests., (Copyright © 2022 Elsevier Inc. All rights reserved.)

The neurotrophin brain-derived neurotrophic factor (BDNF) plays multiple roles in the nervous system, including in neuronal development, in long-term synaptic potentiation in different brain regions, and in neuronal survival. Alterations in these regulatory mechanisms account for several diseases of the nervous system. The synaptic effects of BDNF mediated by activation of tropomyosin receptor kinase B (TrkB) receptors are partly mediated by stimulation of local protein synthesis which is now considered a ubiquitous feature in both presynaptic and postsynaptic compartments of the neuron. The capacity to locally synthesize proteins is of great relevance at several neuronal developmental stages, including during neurite development, synapse formation, and stabilization. The available evidence shows that the effects of BDNF-TrkB signaling on local protein synthesis regulate the structure and function of the developing and mature synapses. While a large number of studies have illustrated a wide range of effects of BDNF on the postsynaptic proteome, a growing number of studies also point to presynaptic effects of the neurotrophin in the local regulation of the protein composition at the presynaptic level. Here, we will review the latest evidence on the role of BDNF in local protein synthesis, comparing the effects on the presynaptic and postsynaptic compartments. Additionally, we overview the relevance of BDNF-associated local protein synthesis in neuronal development and synaptic plasticity, at the presynaptic and postsynaptic compartments, and their relevance in terms of disease. This article is categorized under: RNA Interactions with Proteins and Other Molecules > Protein-RNA Interactions: Functional Implications RNA Export and Localization > RNA Localization., (© 2022 Wiley Periodicals LLC.)

Chick ciliary ganglia (CG) are part of the parasympathetic nervous system and are responsible for the innervation of the muscle tissues present in the eye. This ganglion is constituted by a homogenous population of ciliary and choroidal neurons that innervate striated and smooth muscle fibers, respectively. Each of these neuronal types regulate specific eye structures and functions. Over the years, neuronal cultures of the chick ciliary ganglia were shown to be effective cell models in the study of muscle-nervous system interactions, which communicate through cholinergic synapses. Ciliary ganglion neurons are, in its majority, cholinergic. This cell model has been shown to be useful comparatively to previously used heterogeneous cell models that comprise several neuronal types, besides cholinergic. Anatomically, the ciliary ganglion is localized between the optic nerve (ON) and the choroid fissure (CF). Here, we describe a detailed procedure for the dissection, dissociation and in vitro culture of ciliary ganglia neurons from chick embryos. We provide a step-by-step protocol in order to obtain highly pure and stable cellular cultures of CG neurons, highlighting key steps of the process. These cultures can be maintained in vitro for 15 days and, hereby, we show the normal development of CG cultures. The results also show that these neurons can interact with muscle fibers through neuro-muscular cholinergic synapses.

Gliomas are the most common type of primary brain tumors, presenting high mortality and recurrence rates that highlight the need for the development of more efficient therapies. In that context, we investigated iron(iii) (FeL) and copper(ii) (CuL) complexes containing the tetradentate ligand 2-{[(3-chloro-2-hydroxy-propyl)-pyridin-2-ylmethyl-amino]-methyl}-phenol (L) as potential antimetastatic compounds in glioma cells. These complexes were designed to act as mimetics of antioxidant metalloenzymes (catalases and superoxide dismutase) and thus interfere with the production of reactive oxygen species (ROS), important signaling molecules that have been linked to the induction of Epithelial-Mesenchymal Transition (EMT) in cancer cells, a process associated with cancer invasion and aggressiveness. The results obtained have revealed that, in vitro , both compounds act as superoxide dismutase or catalase mimetics, and this translated in glioma cells into a decrease in ROS levels in FeL-treated cells. In addition, both complexes were found to inhibit the migration of monolayer-grown H4 cells and lead to decreased expression of EMT markers. More importantly, this behavior was recapitulated in 3D spheroids models, where CuL in particular was found to completely inhibit the invasion ability of glioma cells, with or without cellular irradiation with X-rays, which is suggestive of these compounds' potential to be used in combination with radiotherapy. Overall, the results herein obtained describe the novel use of these complexes as agents that are able to interfere with regulation of EMT and the invasive behavior of glioma cells, an application that deserves to be further explored., Competing Interests: There are no conflicts to declare., (This journal is © The Royal Society of Chemistry.)

Ribosomes and a subset of cellular mRNAs are trafficked into axons of developing neurons. The axonal localization of translational machinery allows new proteins to be rapidly and locally synthesized during axonal growth and pathfinding. However, in mature neurons, axonal ribosomes are significantly reduced or even absent. The mechanism that elicits this removal is currently unknown. Here, we demonstrate that synapse formation is the trigger for ribosome reduction in mature axons. In vivo analysis shows that axonal ribosome levels decrease in rat brain at a developmental stage coincident with synapse formation. Next, we observe in vitro that different synaptogenic inducers trigger an overall decrease of ribosomal proteins and rRNA in the axons of spinal motor neurons. We further observe that this process is dependent on the ubiquitin-proteasome system but not on autophagy. Together, these data identify synaptogenesis as the long missing biological trigger that leads to ribosome disappearance during axonal maturation., (Copyright © 2019 The Authors. Published by Elsevier Inc. All rights reserved.)

GABA A receptors (GABA A R) are the major players in fast inhibitory neurotransmission in the central nervous system (CNS). Regulation of GABA A R trafficking and the control of their surface expression play important roles in the modulation of the strength of synaptic inhibition. Different pieces of evidence show that alterations in the surface distribution of GABA A R and dysregulation of their turnover impair the activity of inhibitory synapses. A diminished efficacy of inhibitory neurotransmission affects the excitatory/inhibitory balance and is a common feature of various disorders of the CNS characterized by an increased excitability of neuronal networks. The synaptic pool of GABA A R is mainly controlled through regulation of internalization, recycling and lateral diffusion of the receptors. Under physiological condition these mechanisms are finely coordinated to define the strength of GABAergic synapses. In this review article, we focus on the alteration in GABA A R trafficking with an impact on the function of inhibitory synapses in various disorders of the CNS. In particular we discuss how similar molecular mechanisms affecting the synaptic distribution of GABA A R and consequently the excitatory/inhibitory balance may be associated with a wide diversity of pathologies of the CNS, from psychiatric disorders to acute alterations leading to neuronal death. A better understanding of the cellular and molecular mechanisms that contribute to the impairment of GABAergic neurotransmission in these disorders, in particular the alterations in GABA A R trafficking and surface distribution, may lead to the identification of new pharmacological targets and to the development of novel therapeutic strategies.

Objective: To investigate associations between respiratory capacity, quality of life and cognitive function in elderly individuals., Methods: The sample included 386 elderly individuals (232 women). Respiratory capacity assessment was based on maximal expiratory pressure measured at peak expiratory flow. Subjects were classified according to peak expiratory flow values adjusted for sex, age and height of individuals with normal (peak expiratory flow curve <80% and >60%) or reduced (peak expiratory flow curve < 60%) respiratory capacity. The World Health Organization Quality of Life Questionnaire and the Mini-Mental State Examination were used to assess quality of life and cognitive function, respectively., Results: Elderly women with reduced respiratory capacity scored lower on the Mini-Mental State Examination (p=0.048) and quality of life questionnaire (p=0.040) compared to those with normal respiratory capacity. These differences were not observed in men (p>0.05)., Conclusion: Reduced respiratory capacity was associated with poorer quality of life and cognitive function in elderly women. These associations were not observed in elderly men.

The synthesis, X-ray molecular structure, physico-chemical characterization and dual antioxidant activity (catalase and superoxide dismutase) of a new polymeric mixed valence Mn(III)Mn(II) complex, containing the ligand H 2 BPClNOL (N-(2-hydroxybenzyl)-N-(2-pyridylmethyl)[(3-chloro)(2-hydroxy)] propylamine) is described. The monomeric unit is composed of a dinuclear Mn(II)Mn(III) moiety, [Mn(III)(μ-HBPClNOL)(μ-BPClNOL)Mn(II)(Cl)](ClO 4 )·2H 2 O, 1 , in which the Mn ions are connected by two different bridging groups provided by two molecules of the ligand H 2 BPClNOL, a phenoxide and an alkoxide group. In the solid state, this mixed valence dinuclear unit is connected to its neighbors through chloro bridges. Magnetic measurements indicated the presence of ferromagnetic [ J = +0.076(13) cm -1 ] and antiferromagnetic [ J = -5.224(13) cm -1 ] interactions. The compound promotes O 2 • - dismutation in aqueous solution (IC 50 = 0.370 μmol dm -3 , k cat = 3.6x10 6 M -1 s -1 ). EPR studies revealed that a high-valent Mn(III)-O-Mn(IV) species is involved in the superoxide dismutation catalytic cycle. Complex 1 shows catalase activity only in the presence of a base, e.g., piperazine or triethylamine. Kinetic studies were carried out in the presence of piperazine and employing two different methods, resulting in k cat values of 0.58 ± 0.03 s -1 (detection of O 2 production employing a Clark electrode) and 2.59 ± 0.12 s -1 (H 2 O 2 consuption recorded via UV-Vis). EPR and ESI-(+)-MS studies indicate that piperazine induces the oxidation of 1 , resulting in the formation of the catalytically active Mn(III)-O-Mn(IV) species.

Background: The deficiency in 1α, 25-dihydroxyvitamin D3 (VD3) seems to increase the risk for neurodegenerative pathologies, including Parkinson's disease (PD). The majority of its actions are mediated by the transcription factor, VD3 receptor (VD3R)., Methods: The neuroprotective effects of VD3 were investigated on a PD model. Male Wistar rats were divided into the following groups: sham-operated (SO), 6-OHDA-lesioned (non-treated), and 6-OHDA-lesioned and treated with VD3 (7 days before the lesion, pre-treatment or for 14 days after the 6-OHDA striatal lesion, post-treatment). Afterwards, the animals were subjected to behavioral tests and euthanized for striatal neurochemical and immunohistochemical assays. The data were analyzed by ANOVA and the Tukey test and considered significant for p < 0.05., Results: We showed that pre- or post-treatments with VD3 reversed behavioral changes and improved the decreased DA contents of the 6-OHDA group. In addition, VD3 reduced the oxidative stress, increased (TH and DAT), and reduced (TNF-alpha) immunostainings in the lesioned striata. While significant decreases in VD3R immunoreactivity were observed after the 6-OHDA lesion, these changes were blocked after VD3 pre- or post-treatments. We showed that VD3 offers neuroprotection, decreasing behavioral changes, DA depletion, and oxidative stress. In addition, it reverses partially or completely TH, DAT, TNF-alpha, and VD3R decreases of immunoreactivities in the non-treated 6-OHDA group., Conclusions: Taken together, VD3 effects could result from its anti-inflammatory and antioxidant actions and from its actions on VD3R. These findings should stimulate translational research towards the VD3 potential for prevention or treatment of neurodegenerative diseases, as PD.

Objectives: Aspidosperma species are used for several diseases, especially for malaria in Brazil. Although the genus is object of pharmacological studies, almost none are found on Aspidosperma pyrifolium. We investigate neuroprotective, antioxidant and anti-inflammatory properties of the APSE-Aq fraction (benzoic acid glycosylated derivative) on Parkinson's disease model., Methods: Male Wistar rats were subjected to a 6-hydroxydopamine injection into the right striatum and treated or not with APSE-Aq (100 or 200 mg/kg, p.o.). The sham-operated group was injected with saline. Two weeks later, animals were subjected to behavioural, neurochemical and immunohistochemical evaluation. The data were analysed by ANOVA and Tukey test., Key Findings: The APSE-Aq-treated group shows a partial recovery of behavioural changes as compared with the untreated-6-hydroxydopamine group. A partial recovery was also observed in nitrite contents and lipid peroxidation. APSE-Aq treatments significantly reversed decreases in striatal dopamine and metabolites in the untreated 6-hydroxydopamine group. Immunostainings for markers as tyrosine hydroxylase and dopamine transporter decreased in the untreated 6-hydroxydopamine group and values recovered after APSE-Aq treatments. Similar data were seen for TNF-alpha., Conclusion: APSE-Aq presents neuroprotective, antioxidant and anti-inflammatory activities. Considering that APSE-Aq is chemically related to salicylic acid, it may act on similar targets., (© 2018 Royal Pharmaceutical Society.)

Proneurotrophins were initially thought to be simple inactive precursors, only responsible for promoting the folding of the mature domain and for the regulation of the neurotrophin secretory pathway. However, recent evidence shows that proneurotrophins can be secreted to the extracellular space, bind with high affinity to specific receptor complexes and induce activation of the apoptotic machinery with subsequent cell death of different neuronal populations. These pathways can be activated due to injury and to several neurodegenerative disorders, which promote proneurotrophin secretion to the extracellular space. In addition to neuropathology, extracellular proneurotrophins also play a pivotal role in many other cellular mechanisms in the nervous system. Proneurotrophins were shown to mediate synaptic plasticity, namely long-term depression in hippocampal neurons. They are also important in axonal development, and an increase of pro- to mature neurotrophin ratio has been described as a trigger of cell death. The conversion of proneurotrophins into the respective mature form is controlled by the action of several enzymes and regulators. The failure in this regulation is now considered one of the possible mechanisms responsible for pathological cell death associated to proneurotrophins. Here, we discuss the processes behind proneurotrophin action, with particular focus on proBDNF and proNGF and their regulatory pathways. Additionally, we review the most recent studies concerning proneurotrophin involvement in neuronal death, in several disease-associated states in the CNS and PNS, and discuss future avenues of investigation in the proneurotrophin field.

Valproic acid (VA) is an antiepileptic that is also used for the treatment of bipolar disorders. The objective was to evaluate the neuroprotective effects of VA on a brain ischemia model. The groups of male Wistar rats were: SO (sham-operated), ischemic and ischemic treated with VA (25, 50 and 100 mg/kg, p.o.). After anesthesia with ketamine and xilazine, the animals were subjected to clamping of carotid arteries (30 min) and reperfusion. Except for the carotid clamping, the SO group was submitted to the same procedure. On the 7th day, the animals were behaviorally evaluated, euthanized and had their brain dissected for neurochemical and immunohistochemical assays. The data were analyzed by ANOVA and Tukey as the post hoc test. The results showed that VA reversed partly or completely the behavioral (locomotor activity and memory deficits), neurochemical (striatal DA and DOPAC levels, brain nitrite and lipid peroxidation) and immunohistochemical alterations (iNOS, COX-2, HDAC and GSK3) observed in the untreated ischemic group. VA neuroprotective effects are probably related to its anti-inflammatory and antioxidant properties, as well as to HDAC and GSK3 inhibitory effects. These findings stimulate translational studies focusing on VA as a neuroprotective drug to be potentially used in the clinic for several neurological conditions., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Brain-derived neurotrophic factor (BDNF) is an important mediator of long-term synaptic potentiation (LTP) in the hippocampus. The local effects of BDNF depend on the activation of translation activity, which requires the delivery of transcripts to the synapse. In this work, we found that neuronal activity regulates the dendritic localization of the RNA-binding protein heterogeneous nuclear ribonucleoprotein K (hnRNP K) in cultured rat hippocampal neurons by stimulating BDNF-Trk signaling. Microarray experiments identified a large number of transcripts that are coimmunoprecipitated with hnRNP K, and about 60% of these transcripts are dissociated from the protein upon stimulation of rat hippocampal neurons with BDNF. In vivo studies also showed a role for TrkB signaling in the dissociation of transcripts from hnRNP K upon high-frequency stimulation (HFS) of medial perforant path-granule cell synapses of male rat dentate gyrus (DG). Furthermore, treatment of rat hippocampal synaptoneurosomes with BDNF decreased the coimmunoprecipitation of hnRNP K with mRNAs coding for glutamate receptor subunits, Ca 2+ - and calmodulin-dependent protein kinase IIβ (CaMKIIβ) and BDNF. Downregulation of hnRNP K impaired the BDNF-induced enhancement of NMDA receptor (NMDAR)-mediated mEPSC, and similar results were obtained upon inhibition of protein synthesis with cycloheximide. The results demonstrate that BDNF regulates specific populations of hnRNP-associated mRNAs in neuronal dendrites and suggests an important role of hnRNP K in BDNF-dependent forms of synaptic plasticity.

Ketamine (KET), a NMDA receptor antagonist, has been studied for its rapid and efficacious antidepressant effect, even for the treatment-resistant depression. Although depression is a major cause of disability worldwide, the treatment can be feasible, affordable and cost-effective, decreasing the population health burden. We evaluated the antidepressive-like effects of KET and its actions on monoamine contents (DA and its metabolites, as well as 5-HT) and on tyrosine hydroxylase (TH). In addition DAT and SERT (DA and 5-HT transporters, respectively) were also assessed. Male Swiss mice were divided into Control and KET-treated groups. The animals were acutely treated with KET (2, 5 or 10 mg/kg, i.p.) and subjected to the forced swimming test, for evaluation of the antidepressive-like behavior. Imipramine and fluoxetine were used as references. The results showed that KET decreased dose-dependently the immobility time and shortly after the test, the animals were euthanized for striatal dissections and monoamine determinations. In addition, the brain (striata, hippocampi and prefrontal cortices) was immunohistochemically processed for TH, DAT and SERT. KET at its higher dose increased DA and its metabolites (DOPAC and HVA) and mainly 5-HT contents, in mice striata, effects associated with increases in TH and decreases in DAT immunoreactivities. Furthermore, reductions in SERT immunoreactivities were observed in the striatum and hippocampus. The results indicate that KET antidepressive-like effect probably involves, among other factors, monoaminergic pathways, as suggested by the increased striatal TH immunoreactivity and reduced brain DA (DAT) and 5-HT (SERT) transporters.

This protocol aims at standardizing the procedure to obtain primary cultures of hippocampal and cerebrocortical neurons for in vitro experiments. Cultures should be prepared from cells isolated during embryonic development when neuronal precursor cells are not yet fully differentiated. This helps increasing the quality and quantity of cells, while offering minimal cell death that often occurs during dissociation of differentiated neurons. Cells plated under the appropriate conditions, either in Petri-dishes or in multi-well plates, will develop and establish synaptic contacts over time since the neuronal culture medium provides the nutrients and trophic factors required for differentiation. In this protocol we describe the methodology for the preparation of both cortical and hippocampal neuronal cultures., (Copyright © 2017 The Authors; exclusive licensee Bio-protocol LLC.)

Mesenchymal stem cells (MSCs) have been used for cell-based therapies in regenerative medicine, with increasing importance in central and peripheral nervous system repair. However, MSCs grafting present disadvantages, such as, a high number of cells required for transplantation and low survival rate when transplanted into the central nervous system (CNS). In line with this, MSCs secretome which present on its composition a wide range of molecules (neurotrophins, cytokines) and microvesicles, can be a solution to surpass these problems. However, the effect of MSCs secretome in axonal elongation is poorly understood. In this study, we demonstrate that application of MSCs secretome to both rat cortical and hippocampal neurons induces an increase in axonal length. In addition, we show that this growth effect is axonal intrinsic with no contribution from the cell body. To further understand which are the molecules required for secretome-induced axonal outgrowth effect, we depleted brain-derived neurotrophic factor (BDNF) from the secretome. Our results show that in the absence of BDNF, secretome-induced axonal elongation effect is lost and that axons present a reduced axonal growth rate. Altogether, our results demonstrate that MSCs secretome is able to promote axonal outgrowth in CNS neurons and this effect is mediated by BDNF.

In recent years, signaling through ubiquitin has been shown to be of great importance for normal brain development. Indeed, fluctuations in ubiquitin levels and spontaneous mutations in (de)ubiquitination enzymes greatly perturb synapse formation and neuronal transmission. In the brain, expression of lysine (K) 48-linked ubiquitin chains is higher at a developmental stage coincident with synaptogenesis. Nevertheless, no studies have so far delved into the involvement of this type of polyubiquitin chains in synapse formation. We have recently proposed a role for polyubiquitinated conjugates as triggering signals for presynaptic assembly. Herein, we aimed at characterizing the axonal distribution of K48 polyubiquitin and its dynamics throughout the course of presynaptic formation. To accomplish so, we used an ubiquitination-induced fluorescence complementation (UiFC) strategy for the visualization of K48 polyubiquitin in live hippocampal neurons. We first validated its use in neurons by analyzing changing levels of polyubiquitin. UiFC signal is diffusely distributed with distinct aggregates in somas, dendrites and axons, which perfectly colocalize with staining for a K48-specific antibody. Axonal UiFC aggregates are relatively stable and new aggregates are formed as an axon grows. Approximately 65% of UiFC aggregates colocalize with synaptic vesicle clusters and they preferentially appear in the axonal domains of axo-somatodendritic synapses when compared to isolated axons. We then evaluated axonal accumulation of K48 ubiquitinated signals in bead-induced synapses. We observed rapid accumulation of UiFC signal and endogenous K48 ubiquitin at the sites of newly formed presynapses. Lastly, we show by means of a microfluidic platform, for the isolation of axons, that presynaptic clustering on beads is dependent on E1-mediated ubiquitination at the axonal level. Altogether, these results indicate that enrichment of K48 polyubiquitin at the site of nascent presynaptic terminals is an important axon-intrinsic event for presynaptic differentiation.

Purpose: The aim of this study was to evaluate the effects of green LED light on inflammatory cells in skin burns: a histological study in rats., Methods: In this study, 40 rats were randomly divided into 2 groups: G1 - Control (CTR) and G2 - Green Led (LED). Immediately after injury, green light (60J/cm(2), 10s, λ520 at 550nm) was applied in a timely manner in the four coinciding points of the wound angles and at each point, the amount of 60J/cm (2) with a time of 10s was delivered, totaling 240J/cm(2) per session with 24h intervals until the day before animal sacrifice at 3, 7, 14 and 21 days with a lethal dose of intraperitoneal anesthetic., Results: In the histological analysis, animals treated with green LED, from 7 days, showed a significant decrease (p<0.05) in inflammatory cells when compared to control group., Conclusions: Green LED light provides an anti-inflammatory effect on skin burns of rats., (Copyright © 2015 Elsevier Ltd and ISBI. All rights reserved.)

Oxidative stress and endoplasmic reticulum (ER) stress have been associated with Alzheimer's disease (AD) progression. In this study we analyzed whether oxidative stress involving changes in Nrf2 and ER stress may constitute early events in AD pathogenesis by using human peripheral blood cells and an AD transgenic mouse model at different disease stages. Increased oxidative stress and increased phosphorylated Nrf2 (p(Ser40)Nrf2) were observed in human peripheral blood mononuclear cells (PBMCs) isolated from individuals with mild cognitive impairment (MCI). Moreover, we observed impaired ER Ca2+ homeostasis and increased ER stress markers in PBMCs from MCI individuals and mild AD patients. Evidence of early oxidative stress defense mechanisms in AD was substantiated by increased p(Ser40)Nrf2 in 3month-old 3xTg-AD male mice PBMCs, and also with increased nuclear Nrf2 levels in brain cortex. However, SOD1 protein levels were decreased in human MCI PBMCs and in 3xTg-AD mice brain cortex; the latter further correlated with reduced SOD1 mRNA levels. Increased ER stress was also detected in the brain cortex of young female and old male 3xTg-AD mice. We demonstrate oxidative stress and early Nrf2 activation in AD human and mouse models, which fails to regulate some of its targets, leading to repressed expression of antioxidant defenses (e.g., SOD-1), and extending to ER stress. Results suggest markers of prodromal AD linked to oxidative stress associated with Nrf2 activation and ER stress that may be followed in human peripheral blood mononuclear cells., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Sporotrichosis is a chronic infectious disease affecting both humans and animals. For many years, this subcutaneous mycosis had been attributed to a single etiological agent; however, it is now known that this taxon consists of a complex of at least four pathogenic species, including Sporothrix schenckii and Sporothrix brasiliensis. Gp70 was previously shown to be an important antigen and adhesin expressed on the fungal cell surface and may have a key role in immunomodulation and host response. The aim of this work was to study the virulence, morphometry, cell surface topology and gp70 expression of clinical isolates of S. brasiliensis compared with two reference strains of S. schenckii. Several clinical isolates related to severe human cases or associated with the Brazilian zoonotic outbreak of sporotrichosis were genotyped and clustered as S. brasiliensis. Interestingly, in a murine subcutaneous model of sporotrichosis, these isolates showed a higher virulence profile compared with S. schenckii. A single S. brasiliensis isolate from an HIV-positive patient not only showed lower virulence but also presented differences in cell morphometry, cell wall topography and abundant gp70 expression compared with the virulent isolates. In contrast, the highly virulent S. brasiliensis isolates showed reduced levels of cell wall gp70. These observations were confirmed by the topographical location of the gp70 antigen using immunoelectromicroscopy in both species. In addition, the gp70 molecule was sequenced and identified using mass spectrometry, and the sequenced peptides were aligned into predicted proteins using Blastp with the S. schenckii and S. brasiliensis genomes.

Gardner's syndrome, an autosomal dominant syndrome, is linked to familial adenomatosis polyposis (FAP), which is known mainly as a colorectal disease. FAP also presents extracolonically as intestinal polyposis, multiple osteomas, cutaneous cysts, or fibromas. This article reports the case of a 66-year-old white woman who was referred to the Oral Medicine Clinic, School of Dentistry, Universidade Vale do Rio Verde, Brazil, for evaluation of multiple sclerotic, asymptomatic masses in the jaws that were observed in a routine periapical radiographic exam by a dentist. The patient presented with intestinal poliposis, periosteal osteoma in the face, and fibromas and multiple endosteal osteomas in the maxilla, which are indications of Gardner's syndrome. The clinical differential diagnosis included multiple buccal exostoses, idiopathic osteosclerosis, cemento-osseous dysplasias, multiple odontomas, osteomas, and Gardner's syndrome. Patients with a suspected diagnosis of Gardner's syndrome should be referred to a dermatologist, have a colonoscopy performed, and be followed up by a dentist.

It was recently established that the stomach-derived ghrelin and the adipokine leptin promote learning and memory through actions within the hippocampus. Changes in the peripheral or brain levels of these peptides were described in Alzheimer's disease (AD) patients and were shown to correlate with the severity of cognitive decline. Furthermore, in vivo and in vitro studies demonstrated that leptin or ghrelin can ameliorate amyloid and tau pathologies as well as cognitive deficits. However, the exact role of these peptides in AD is far from being elucidated. To fill this gap, our working hypothesis was that leptin and ghrelin can exert a neuroprotective role in AD suppressing hippocampal dysfunction triggered by synapto- and neurotoxic amyloid-β oligomers (AβO). Using primary cultured hippocampal neurons, we demonstrated that both peptides reduce AβO-induced production of superoxide and mitochondrial membrane depolarization, improving cell survival, and inhibit cell death through a receptor-dependent mechanism. Furthermore, it was shown that in AβO-treated neurons both leptin and ghrelin prevent glycogen synthase kinase 3β activation. Therefore, the evidence gathered in this study revealed that leptin and ghrelin can act as neuroprotective agents able to rescue hippocampal neurons from AβO toxicity, thus highlighting their potential therapeutic role in AD., (Copyright © 2013 IBRO. Published by Elsevier Ltd. All rights reserved.)

Potassium iodide, as a saturated solution, is a valuable drug in the dermatologist's therapeutic arsenal and is useful for the treatment of different diseases due to its immunomodulatory features. However, its prescription has become increasingly less frequent in dermatology practice. Little knowledge about its exact mechanism of action, lack of interest from the pharmaceutical industry, the advent of new drugs, and the toxicity caused by the use of high doses of the drug are some possible explanations for that. Consequently, there are few scientific studies on the pharmacological aspects, dosage and efficacy of this drug. Also, there is no conventional standard on how to manipulate and prescribe the saturated solution of potassium iodide, which leads to unawareness of the exact amount of the salt being delivered in grams to patients. Considering that dosage is directly related to toxicity and the immunomodulatory features of this drug, it is essential to define the amount to be prescribed and to reduce it to a minimum effective dose in order to minimize the risks of intolerance and thus improve treatment adherence. This review is relevant due to the fact that the saturated solution of potassium iodide is often the only therapeutic choice available for the treatment of some infectious, inflammatory and immune-mediated dermatoses, no matter whether the reason is specific indication, failure of a previous therapy or cost-effectiveness.

Previously we reported that amyloid-β (Aβ) leads to endoplasmic reticulum (ER) stress in cultured cortical neurons and that ER-mitochondria Ca(2+) transfer is involved in Aβ-induced apoptotic neuronal cell death. In cybrid cells which recreate the defect in mitochondrial cytochrome c oxidase (COX) activity observed in platelets from Alzheimer's disease (AD) patients, we have shown that mitochondrial dysfunction affects the ER stress response triggered by Aβ. Here, we further investigated the impact of COX inhibition on Aβ-induced ER dysfunction using a neuronal model. Primary cultures of cortical neurons were challenged with toxic concentrations of Aβ upon chemical inhibition of COX with potassium cyanide (KCN). ER Ca(2+) homeostasis was evaluated under these conditions, together with the levels of ER stress markers, namely the chaperone GRP78 and XBP-1, a mediator of the ER unfolded protein response (UPR). We demonstrated that COX inhibition potentiates the Aβ-induced depletion of ER Ca(2+) content. KCN pre-treatment was also shown to enhance the rise of cytosolic Ca(2+) levels triggered by Aβ and thapsigargin, a widely used ER stressor. This effect was reverted in the presence of dantrolene, an inhibitor of ER Ca(2+) release through ryanodine receptors. Similarly, the increase in GRP78 and XBP-1 protein levels was shown to be higher in neurons treated with Aβ or thapsigargin in the presence of KCN in comparison with levels determined in neurons treated with the neurotoxins alone. Although the decrease in cell survival, the activation of caspase-9- and -3-mediated apoptotic cell death observed in Aβ- and thapsigargin-treated neurons were also potentiated by KCN, this effect is less pronounced than that observed in Ca(2+) signalling and UPR. Furthermore, in neurons treated with Aβ, the potentiating effect of the COX inhibitor in cell survival and death was not prevented by dantrolene. These results show that inhibition of mitochondrial COX activity potentiates Aβ-induced ER dysfunction and, to a less extent, neuronal cell death. Furthermore, data supports that the effect of impaired COX on Aβ-induced cell death occurs independently of Ca(2+) release through ER ryanodine receptors. Together, our data demonstrate that mitochondria dysfunction in AD enhances the neuronal susceptibility to toxic insults, namely to Aβ-induced ER stress, and strongly suggest that the close communication between ER and mitochondria can be a valuable future therapeutic target in AD., (Copyright © 2012 Elsevier Inc. All rights reserved.)

Azidothymidine (AZT) is an antiretroviral drug that affects cell proliferation, apoptosis, and the NF-κB pathway. As multiple myeloma (MM) presents with constitutive activation of NF-κB, we analyzed the effect of AZT on human MM cell lines. We evaluated the cytotoxic effect of AZT in human MM cell lines sensitive (8226/S) or resistant to doxorubicin (8226/DX5) and human T cell lymphoblast-like cells, uterine sarcoma cells, and HUVEC using MTT assay. Cytotoxicity was also evaluated in vivo in nude mice xenografted with 8226/S tumor. The effect of AZT on the expression of genes involved in cell proliferation, apoptosis, angiogenesis, and the NF-κB pathway was analyzed in the xenografts using real-time polymerase chain reaction. AZT was effective against both 8226/S and 8226/DX5 cells in a dose and time-dependent manner (p = 0.02) in vitro and promoted cell cycle arrest in S phase in these cells. The tumor volume was lower in mice treated with AZT compared to untreated mice (p = 0.0003). AZT down-regulated the pro-proliferative genes encoding AKT1, MYC, STAT1, MAPK8, MAPK9, CCL-3, Bcl-3, and cyclin D2; pro-angiogenenic genes encoding VEGF and IL8; and genes involved in cell adhesion (ICAM1 and FN1) and the NF-κB pathway. AZT up-regulated the expression of tumor suppressor gene FOXP1 and the pro-apoptotic genes encoding BID, Bcl-10, and caspase-8. Thus, we demonstrated the cytotoxic effect of AZT in human MM cell lines for the first time. Our data may provide the rationale for future clinical trials of AZT for treating MM.

Alzheimer's disease (AD) is a progressive neurodegenerative disorder affecting both the hippocampus and the cerebral cortex. Reduced synaptic density that occurs early in the disease process seems to be partially due to the overactivation of N-methyl-d-aspartate receptors (NMDARs) leading to excitotoxicity. Recently, we demonstrated that amyloid-beta oligomers (AβO), the species implicated in synaptic loss during the initial disease stages, induce endoplasmic reticulum (ER) stress in cultured neurons. Here, we investigated whether AβO trigger ER stress by an NMDAR-dependent mechanism leading to neuronal dysfunction and analyzed the contribution of GluN2A and GluN2B subunits of this glutamate receptor. Our data revealed that AβO induce ER stress in mature hippocampal cultures, activating ER stress-associated sensors and increasing the levels of the ER chaperone GRP78. We also showed that AβO induce NADPH oxidase (NOX)-mediated superoxide production downstream of GluN2B and impairs ER and cytosolic Ca2+ homeostasis. These events precede changes in cell viability and activation of the ER stress-mediated apoptotic pathway, which was associated with translocation of the transcription factor GADD153 / CHOP to the nucleus and occurred by a caspase-12-independent mechanism. Significantly, ER stress took place after AβO interaction with GluN2B subunits. In addition, AβO-induced ER stress and hippocampal dysfunction were prevented by ifenprodil, an antagonist of GluN2B subunits, while the GluN2A antagonist NVP-AAM077 only slightly attenuated AβO-induced neurotoxicity. Taken together, our results highlight the role of GluN2B subunit of NMDARs on ER stress-mediated hippocampal dysfunction caused by AβO suggesting that it might be a potential therapeutic target during the early stages of AD., (© 2012 The Authors. Aging Cell © 2012 Blackwell Publishing Ltd/Anatomical Society of Great Britain and Ireland.)

Objectives: The aim of this retrospective study was to investigate the results of T-cell large granular lymphocytic leukemia treatment with fludarabine by assessing the complete hematologic response, the complete molecular response, progression-free survival, and overall survival., Methods: We evaluated the records of six patients with T-cell large granular lymphocytic leukemia who were treated with fludarabine as a first-, second-, or third-line therapy, at a dose of 40 mg/m², for three to five days per month and 6 to 8 cycles., Results: Of the six patients investigated with T-cell large granular lymphocytic leukemia who were treated with fludarabine, five (83.3%) were female, and their median age was 36.5 years (range 18 to 73). The median lymphocyte level was 3.4 x 10(9)/L (0.5 to 8.9). All patients exhibited a monoclonal T-cell receptor gamma gene rearrangement at diagnosis. Two (33.3%) patients received fludarabine as first-line treatment, two (33.3%) for refractory disease, one (16.6%) for relapsed disease after the suspension of methotrexate treatment due to liver toxicity, and one (16.6%) due to dyspepsia. A complete hematologic response was achieved in all cases, and a complete molecular response was achieved in five out six cases (83.3%). During a mean follow-up period of 12 months, both the progression-free survival and overall survival rates were 100%., Conclusion: T-cell large granular lymphocytic leukemia demonstrated a high rate of complete hematologic and molecular response to fludarabine, with excellent compliance and tolerability rates. To confirm our results in this rare disease, we believe that fludarabine should be tested in clinical trials as a first-line treatment for T-cell large granular lymphocytic leukemia.

Previously we reported that endoplasmic reticulum (ER)-mitochondria crosstalk is involved in amyloid-β (Aβ)-induced apoptosis. Now we show that mitochondrial dysfunction affects the ER stress response triggered by Aβ using cybrids that recreate the defect in mitochondrial cytochrome c oxidase (COX) activity detected in platelets from Alzheimer's disease (AD) patients. AD and control cybrids were treated with Aβ or classical ER stressors and the ER stress-mediated apoptotic cell death pathway was accessed. Upon treatment, we found increased glucose-regulated protein 78 (GRP78) levels and caspase-4 activation (ER stress markers) which were more pronounced in AD cybrids. Treated AD cybrids also exhibited decreased cell survival as well as increased caspase-3-like activity, poli-ADP-ribose-polymerase (PARP) levels and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL)-positive apoptotic cells. Finally, we showed that Aβ-induced caspase-3 activation in both cybrid cell lines was prevented by dantrolene, thus implicating ER Ca(2+) release in ER stress-mediated apoptosis. Our results demonstrate that mitochondrial dysfunction occurring in AD patients due to COX inhibition potentiates cell susceptibility to Aβ-induced ER stress. This study further supports the close communication between ER and mitochondria during apoptosis in AD., (Copyright © 2012 Elsevier Inc. All rights reserved.)

Alzheimer's disease (AD) is the most common cause of dementia in the elderly, affecting several million of people worldwide. Pathological changes in the AD brain include the presence of amyloid plaques, neurofibrillary tangles, loss of neurons and synapses, and oxidative damage. These changes strongly associate with mitochondrial dysfunction and stress of the endoplasmic reticulum (ER). Mitochondrial dysfunction is intimately linked to the production of reactive oxygen species (ROS) and mitochondrial-driven apoptosis, which appear to be aggravated in the brain of AD patients. Concomitantly, mitochondria are closely associated with ER, and the deleterious crosstalk between both organelles has been shown to be involved in neuronal degeneration in AD. Stimuli that enhance expression of normal and/or folding-defective proteins activate an adaptive unfolded protein response (UPR) that, if unresolved, can cause apoptotic cell death. ER stress also induces the generation of ROS that, together with mitochondrial ROS and decreased activity of several antioxidant defenses, promotes chronic oxidative stress. In this paper we discuss the critical role of mitochondrial and ER dysfunction in oxidative injury in AD cellular and animal models, as well as in biological fluids from AD patients. Progress in developing peripheral and cerebrospinal fluid biomarkers related to oxidative stress will also be summarized.

Five cases of sporotrichosis occurring in pregnant women in a zoonotic epidemic in Rio de Janeiro, Brazil, are described. The main clinical features, as well as the challenging therapeutic choices for this specific group of patients, are discussed.

A case of a follicular lymphoma transformed into a CD20(+) is described which progressed with the loss of CD20 expression after 8 cycles of R-CHOP. This phenomenon is not a rare event and has shown poor prognosis. Our purposes are to describe this event and suggest biopsy in relapsed or progressive disease.

This report shows images of a case of subcutaneous phaeohyphomycosis caused by Exophiala jeanselmei in a patient who has undergone renal transplantation. The paper comments briefly on the disease and emphasizes the need to take this mycosis into account in the differential diagnosis of other dermatoses, including non-infectious dermatoses.

Amyloid-beta (Abeta) peptide plays a significant role in the pathogenesis of Alzheimer's disease (AD). Previously we found that Abeta induces both mitochondrial and endoplasmic reticulum (ER) dysfunction leading to apoptosis, and now we address the relevance of ER-mitochondria crosstalk in apoptotic cell death triggered by Abeta peptide. Using mitochondrial DNA-depleted rho0 cells derived from the human NT2 teratocarcinoma cell line, characterized by the absence of functional mitochondria, and the parental rho+ cells, we report here that treatment with the synthetic Abeta1-40 peptide, or the classical ER stressors thapsigargin or brefeldin A, increases GRP78 expression levels and caspase activity, two ER stress markers, and also depletes ER calcium stores. Significantly, we show that the presence of functional mitochondria is required for ER stress-mediated apoptotic cell death triggered by toxic insults such as Abeta. We found that the increase in the levels of the pro-apoptotic transcription factor GADD153/CHOP, which mediates ER stress-induced cell death, as well as caspase-9 and -3 activation and increased number of TUNEL-positive cells, occurs in treated parental rho+ cells but is abolished in rho0 cells. Our results strongly support the close communication between ER and mitochondria during apoptotic cell death induced by the Abeta peptide and provide insights into the molecular cascade of cell death in AD.

Sporotrichosis is a generally cutaneous, granulomatous, chronic and benign infection. Less frequently the disease may affect the joints. Articular involvement is usually characterized by monoarthritis in the absence of systemic symptoms, generally preceded by skin lesions, and frequently affects immunosuppressed individuals. We describe here the case of a healthy patient presenting knee arthritis without skin lesions, diagnosed as sporotrichosis, and treated with oral itraconazole. Serology used in this case was an invaluable tool for the diagnosis of sporotrichosis arthritis lacking skin lesions.