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Background: Adult patients afflicted with Acute Lymphoblastic Leukemia (ALL) still represent a subset of poor prognosis, with a 5-year overall survival of around 46%. The development of new therapeutic approaches is of utmost importance to the oncologic routine and the search for targeted therapies is ever growing in the translational field. Objective: In this study, we aim to investigate an agonistic effect of poly-ADP-ribose polymerase 1 (PARP1) in inducing parthanatos, a PARP1-dependent and programed mode of cell death, in leukemic cell lines overexpressing PARP1, through inhibition of its antagonist, poly-ADP-ribose glicohydrolase (PARG). Methods and materials: We screened 6 cell lines for their PARP1 mRNA expression through qPCR. Inhibition of their metabolic activity was screened through Alamar Blue assay (Invitrogen™) after treatment with either a PARG inhibitor, JA2131, a PARP inhibitor, AZD2461, or a cytotoxic control in Doxorubicin. An IC50 that considered only cell death was assayed through treatment in serial dilutions followed by DAPI solution staining and analysis in flow cytometry. The profile of early death was determined by co-staining of treated cells with CellEvent™Caspase-3/7 Green Reagent (Invitrogen™), APC Annexin V (BD Pharmingen™) and DAPI Solution. Results and discussion: Of the initially screened cell lines, only B-ALL models significantly overexpresed PARP1 when compared to PBMC of healthy donors. Also, while all models responded to Doxorubicin treatment, only B-ALL models had measurable metabolic inhibition when incubated with PARGi and PARPi treatment. Cell death measured by DAPI has only been conducted in one cell line as of this moment, SUP-B15, a B-ALL model. While PARPi and Doxorubicin had IC50 values similar to those observed in metabolic inhibition by Alamar Blue, PARGi only induced cell death in treatment values approximately 3-fold higher than the necessary for metabolic inhibition. Nevertheless, cell death induced by the treatments demonstrate different early death profiles, with PARGi treatment not inducing any effector caspase activity previously to cell death, a hallmark of parthanatos, while PARPi and Doxorubicin had clear increase in caspase activity and annexin-V staining in early apoptotic cells. Conclusion: Targeted therapeutics with PARG or PARP inhibitors may be a feasible option for B-ALL treatment, with different modes of cell death induction. However, further analysis needs to be conducted in order to better characterize treatment-induced phenotypes and their cytotoxicity over non-neoplastic cells.

Acute Lymphoblastic Leukemia (ALL) is a hematological neoplasm characterized by clonal proliferation of lymphoblasts, being one of the most common types of leukemia in children and young adults, with a worse prognosis when manifested in adult patients. Despite advances in treatment, overall survival (OS) varies widely, influenced by several risk factors, such as cytogenetic characteristics, age, and disease subtype. This study seeks to analyze the impact of these factors on the overall survival of ALL patients, highlighting the importance of a personalized therapeutic approach to improve clinical outcomes. This retrospective study included 53 patients diagnosed with ALL treated at the Hospital Geral de Fortaleza (HGF), which is the largest oncohematology outpatient clinic in the state of Ceará. Patients were stratified according to the World Health Organization (WHO) guideline. The OS was assessed using Kaplan-Meier curves for each risk group and its’ time was defined as the period from diagnosis to death or the last follow-up. Statistical analyses were performed to compare the survival curves between the different risk groups, using the log-rank test to determine statistical significance. Of the 53 analyzed patients, 45 had B-ALL (median age of 40 years) and 8 had T-ALL (median age of 28 years). In all, 23 patients were considered to have high-risk cytogenetic and 30 were over 35 years old. The analysis showed that patients without high-risk cytogenetics (20% in 40 months) had a significantly lower survival compared to those with high-risk cytogenetics (50% in 40 months). This may seem counterintuitive, as high-risk features are expected to be associated with a worse prognosis. Perhaps this is due to the fact that patients with high-risk cytogenetics can receive more intensive or innovative treatments, leading to a better initial response and, consequently, longer survival. Age proved to be an important prognostic factor, with older patients having significantly lower survival (25% in 40 months). This is consistent with the literature, where it is known that advanced age is a risk factor for a worse prognosis in ALL, due to factors such as higher rates of comorbidities and resistance to treatments. Overall, we observed that patients with T-ALL showed slightly better survival compared to patients with B-ALL, particularly in the first 10 months. This may be explained because T-ALL may have a more favorable initial response to chemotherapy treatment compared to B-ALL. This study's results underline the importance of a personalized approach in the ALL treatment. It is evident that factors such as cytogenetics, age, and ALL subtype have a significant impact on patient overall survival. Stratification based on these factors can help in the selection of more appropriate treatments and in the definition of more accurate prognosis.

Acute myeloid leukemia (AML) is a hematologic malignancy characterized by the rapid proliferation of abnormal myeloid cells in the bone marrow and peripheral blood. Cytogenetics plays a crucial role in the risk stratification and prognosis of AML patients, according to the latest guidelines from the World Health Organization (WHO) and the European LeukemiaNet (ELN). This study aims to analyze the impact of high-risk cytogenetics on the survival of non-promyelocytic AML (non-APL) patients. In all, 93 non-APL AML patients were diagnosed between July 2021 and June 2024 at the Hospital Geral de Fortaleza (HGF), which is the largest oncohematology outpatient clinic in the state of Ceará. Survival data from non-APL AML patients were analyzed, stratified according to the presence or absence of high-risk cytogenetics, according to the guidelines. The analysis was performed using the Kaplan-Meier method, comparing two groups: patients with high-risk cytogenetics and patients without high-risk cytogenetics. Among the 93 patients, 47 were women and 46 were men, with a median age of 52.9 years (17-96 years). It was not possible to stratify all patients due to the lack of cytogenetic results. Among the available results, it was possible to stratify 15 patients with high-risk cytogenetic and 38 without high-risk cytogenetic. Herewith, a significant difference in the probability of survival was observed between the two groups over time, where the group with high-risk cytogenetics demonstrated a sharp drop in the probability of survival in the first 10 months, with a very low residual survival after this period, while the group that was not high-risk had a higher and more stable survival rate over time, with a probability of survival greater than 75% at 40 months. The results indicate that the presence of high-risk cytogenetics is associated with a significant worsening in overall survival of non-APL AML patients. Patients with high-risk cytogenetics show a rapid and marked decrease in the probability of survival in the first 10 months after diagnosis, underscoring the need for more aggressive and personalized therapeutic strategies for this subgroup. This occurs similarly in the literature that points to adverse cytogenetic abnormalities, such as in acute myeloid leukemia and chronic lymphocytic leukemias. These findings reinforce the importance of cytogenetic stratification in the clinical AML management. Patients with high-risk cytogenetics have a worse survival, which warrants more intensive therapeutic interventions and close monitoring. Cytogenetics-based risk stratification should be standard practice in treatment assessment and planning.

Objetivo: A Leucemia Linfoblástica Aguda (LLA) é uma neoplasia do sistema hematopoiético, caracterizada pelo acometimento de células progenitoras linfoides, originando a transformação e proliferação clonal anormal de células imaturas, denominadas de blastos. Com o grande acúmulo de blastos, a medula óssea fica sobrecarregada impedindo a maturação de outras linhagens celulares sanguíneas normais. A telomerase humana (hTERT) foi identificada como marcador comum e importante do câncer, pois dados mostram que esta desempenha um papel crítico na proliferação de células aberrantes, no progresso tumoral e na imortalidade da maioria dos tumores. No modelo da LLA, as células do tumor utilizam a telomerase para primeiramente adquirir um fenótipo imortalizado, impedindo a apoptose celular induzida pela redução de telômeros e proporcionando, assim, a sua capacidade de se auto renovar. O splicing alternativo de RNA afeta a maioria dos genes, expandindo a capacidade de codificação e regulando a abundância de isoformas gênicas. Metodologia: Foram utilizados dados de expressão de medula óssea normal importados do Genotype-Tissue Expression (GTEX) e de tecidos neoplásicos, especificamente tecidos com LLA provenientes do TARGET (Therapeutically Applicable Research to Generate Effective Treatments). Esses dados são integrados ao The Cancer Genome Atlas (TCGA) e analisados utilizando a plataforma de exploração e visualização de dados genômicos UCSC Xena. Resultado: Observa-se que existem diferenças de expressão de isoformas do gene hTERT potencialmente reguladas em LLA em comparação com medula óssea. Avaliadas a partir da densidade e TPM, que são formas de visualizar a expressão gênica, permitindo a comparação precisa entre diferentes isoformas em condições experimentais. Essa métrica auxilia a identificar diferenças na expressão gênica das isoformas que podem ser relevantes para o desenvolvimento e progressão da LLA. Discussão: A reativação da telomerase é o principal mecanismo de manutenção dos telômeros nas neoplasias, limitada apenas pela taxa do componente enzimático. Na LLA essa reativação de expressão gênica da hTERT também ocorre, visto em pacientes tanto pediátricos quanto adultos. A regulação e as funções das isoformas de hTERT, devido ao splicing alternativo, ainda não são totalmente compreendidas. Sendo assim, o splicing alternativo de hTERT pode influenciar a regulação da telomerase ativa, com algumas isoformas sendo ineficazes na reposição dos telômeros. Portando o splicing de hTERT em LLA e isoformas não codificantes pode ser uma nova estratégia de inibição da telomerase para impedir a proliferação celular e a sobrevivência das células neoplásicas. Conclusão: Relevante para futuros estudos específicos observando os padrões de expressão de isoformas de hTERT relacionados com a LLA. O trabalho reforça a necessidade de investigações específicas delas na LLA visto que existem estudos que observaram a hiperexpressão gênica da hTERT tanto em linhagens celulares representativas para leucemias agudas quanto em pacientes diagnosticados com LLA. O resumo fornece caminhos e traz à luz as atuais limitações técnicas das análises bioinformáticas da expressão de isoformas de hTERT com foco na LLA.

Objetivo: A leucemia mieloide aguda (LMA) é uma doença geneticamente diversa. Existem vias intracelulares importantes na leucemogênese e manutenção da leucemia, destacando-se a Janus Quinase-Transdutores de Sinal e Ativadores da Transcrição (JAK-STAT). A via JAK-STAT é crucial para a proliferação e sobrevivência das células neoplásicas, podendo contribuir para a resistência a quimioterápicos e desempenhando um papel significativo na patogênese de várias doenças mediadas principalmente por fatores inflamatórios. Mutações somáticas de ganho de função no gene JAK2, como JAK2-V617F, estão amplamente associadas a distúrbios mieloproliferativos, como na LMA. A ativação do fator de transcrição NF-κB induz a produção excessiva de IL-6, regulando positivamente a via de sinalização JAK2-STAT3. O galato de epigalocatequina (EGCG), um polifenol do chá verde (Camellia sinensis), demonstrou efeitos antineoplásicos contra diversas células neoplásicas in vitro. Estudos sobre fitoterápicos revelam resultados promissores, especialmente nas atividades anti-inflamatórias e antitumorais. A inibição pelo EGCG ocorre principalmente através da inibição de NF-κB, que regula a via JAK2. Além disso, o EGCG pode inibir diretamente a via de sinalização da mutação JAK2 V617F. Metodologia: A estrutura experimental da proteína JAK2-V617F (6D21) foi exportado do servidor Protein Data Bank (PDB) e o ligante Galato de Epigalocatequina (EGCG) foi exportado do servidor PUBchem (CID 65064), ambos foram devidamente tratados utilizando os softwares Pymol e Autodocktools. Resultado: Após a realização do docking com o AutoDock Vina, foi observada uma forte afinidade energética de -9,2 kcal/mol. As análises realizadas com o software DiscoveryStudio revelaram várias interações de van der Waals, além de ligações pi-pi e ligações hidrofóbicas com os resíduos LYS581 e ILE559, evidenciando uma estabilidade do complexo ligante-proteína. Discussão: Os inibidores de JAK (JAKinibis) são uma classe de medicamentos projetados para bloquear a sinalização da via JAK-STAT. A via JAK2-STAT3 é frequentemente afetada em neoplasias, sendo crucial para a proliferação celular, diferenciação e evasão da apoptose das células neoplásicas. Também está envolvida na iniciação e progressão de respostas inflamatórias e imunológicas em diversos processos patológicos, sendo importante na manutenção da leucemia. A produção excessiva de IL-6 ocorre em resposta à ativação inflamatória pelo NF-κB, que regula positivamente a via JAK2-STAT3. A ativação do fator de transcrição NF-κB induz a expressão de IL-6, seguida pela ativação de vias de sinalização subsequentes. Alterações nos fatores de transcrição podem desempenhar um papel crucial na malignidade, progressão, invasão, metástase e resistência a quimioterápicos. Portanto, o bloqueio da JAK2 V617F, que ativa indefinidamente a sinalização da via, e a inibição da via inflamatória por NF-κB pelo EGCG, pode ser uma abordagem promissora na regulação da neoplasia. Conclusão: A ativação da JAK2 está diretamente ligada a características celulares desfavoráveis para pacientes com LMA. Assim, a inibição e regulação desta via apresenta-se como uma abordagem promissora para o tratamento e melhoria do prognóstico de neoplasias. Nesse sentido, nossas análises mostram que o EGCG pode ter efeito promissor na inibição de JAK2-V617F, favorecendo a inibição de vias inflamatórias melhorando o prognóstico de pacientes leucêmicos.

Objetivo: A mutação JAK2-V617F é prevalente nas neoplasias mieloproliferativas (MPNs), podendo evoluir para leucemia mieloide aguda (LMA) de forma secundária com resistência ao tratamento. Marcada por mutações somáticas detectáveis em quase 98% dos casos, a LMA é caracterizada pela proliferação descontrolada de células hematopoiéticas, resultando em um acúmulo de células mieloides imaturas e anormais. Por ser geneticamente instável, o quadro dos pacientes com LMA é influenciado por alterações em vias importantes para a fisiologia da doença. Dentre elas, estão as mutações na via Janus Quinase-Transdutores de Sinal e Ativadores da Transcrição (JAK-STAT). JAK-STAT é crucial para a proliferação e sobrevivência celular, contribuindo para a resistência aos medicamentos. A mutação JAK2-V617F ocorre por uma substituição de guanina por timina no nucleotídeo 1849 do éxon 14, resultando na troca do aminoácido valina por fenilalanina na posição 617 da cadeia polipeptídica (V617F) ativando continuamente a JAK2, aumentando a sensibilidade a fatores de crescimento, a proliferação e manutenção neoplásica, promovendo a sobrevivência e crescimento celular. Nesse sentido, a busca por terapias que visem a inibição de JAK2-V617F pode superar a resistência aos tratamentos e melhorar os resultados clínicos em pacientes leucêmicos. Sendo assim, neste trabalho, buscamos observar o efeito do Kaempferol (KPL), um flavonoide encontrado em muitos produtos naturais, na proteína na JAK2-V617F. Metodologia: A estrutura da proteína JAK2-V617F (6D21) foi exportada do Protein Data Bank (PDB) e a estrutura do ligante KPL (CID 5280863) foi exportada do PUBchem, ambos tratados utilizando os softwares Pymol e Autodocktools. O Docking Molecular e as análises foram realizadas utilizando o software Autodock Vina e o software Discovery Studio. Resultado: Nossos resultados mostraram afinidade energética de -8.5 kcal/mol e após análises, observou-se uma série de interações de van der Waals, interações pi-pi e hidrofóbicas com os resíduos SER633 e ILE559, gerando assim, estabilidade do complexo ligante-proteína. Discussão: A JAK-STAT está diretamente ligada a prognósticos desvantajosos em doenças oncohematológicas. Nesse sentido, pesquisas observaram uma diminuição da proliferação de células KG1a de LMA por KPL, possivelmente por estar relacionado à inibição da via de sinalização JAK2-STAT3. Ademais, segundo diversos trabalhos o KPL é um anticancerígeno promissor e que pode favorecer a supressão da angiogênese, estimulação da apoptose e parada do ciclo celular. Conclusão: A busca por tratamentos mais eficazes para pacientes JAK2-V617F-positivos é constante, devido à associação dessa ativação com características celulares desfavoráveis. Embora existam compostos que interrompem a sinalização JAK-STAT pela inibição de JAK, sua eficácia em JAK2-V617F é limitada. Portanto, novas estratégias contra doenças oncohematológicas com JAK2-V617F-positivas são necessárias. Sendo assim, nossas análises mostram que o KPL apresenta um efeito inibidor promissor na mutação JAK2-V617F, contribuindo para a inibição da via, diminuindo a proliferação celular e melhorando o prognóstico de pacientes com LMA.

Objetivo: O ciclo circadiano (CC) é um sistema biológico fundamental que regula vários processos fisiológicos para manter a homeostase, adaptando-se a mudanças externas, como o ciclo claro/escuro. Este sistema, orquestra a expressão gênica envolvida no metabolismo, proliferação, reparo de DNA, respostas ambientais e imunidade. Os CLOCK GENES (CGs), CLOCK e BMAL1, que formam heterodímeros CLOCK-BMAL1 são componentes chave para esse ciclo, impulsionando e expressão de outros CG's e sendo auto regulados por eles, por meio de loops de feedback. As interrupções no ritmo circadiano estão ligadas a várias doenças, incluindo distúrbios metabólicos, condições neurodegenerativas e em diversas neoplasias. Ademais, os genes CLOCK e BMAL1 chamam a atenção já que foram observados expresso negativamente em todos os tipos de leucemias. Sendo assim, buscamos avaliar os papéis subexplorados da proteína CLOCK-interacting pacemaker, Circadian (CIPC), um regulador negativo independente do complexo CLOCK-BMAL1, e suas implicações no câncer, particularmente em leucemias. Ademais, são poucos os trabalhos que relatam as atividades moleculares de CIPC, porém já é sabido que essa proteína tem a capacidade de inibir a atividade do complexo CLOCK, regulando negativamente a transcrição de genes controlados pelo CC. Isso afeta a expressão de vários genes que são importantes para a manutenção dos ritmos circadianos em diversos tecidos, porém há uma falta de trabalhos que avaliam a capacidade oncológica de CIPC. Metodologia: Foram utilizados dados de expressão de tecidos normais importados do Genotype-Tissue Expression (GTEX) e neoplásicos, integrados ao The Cancer Genome Atlas (TCGA), a partir da plataforma de exploração e visualização de dados genômicos UCSC Xena. Os dados foram organizados e as análises de estatísticas foram realizadas utilizando o software Graphpad Prism versão 8.0.1. Para a análise estatística, foram aplicados os testes de normalidade de Shapiro-Wilk seguido do teste comparativo de Mann-Whitney. Resultado: A análise da expressão gênica apresentou um valor p significativo (p < 0,0001), demonstrando que o CIPC é hiperexpresso na leucemia mieloide aguda (LMA) em comparação com tecidos saudáveis. Discussão: Estudos anteriores mostraram que CLOCK-BMAL1 está negativamente expresso em todos os tipos de leucemia, podendo estar associados a características desfavoráveis, como resistência e pior prognóstico, nesse sentido o papel de regulador negativo do CC de CIPC poderá está relacionado a leucemias, favorecendo a depressão circadiana levando também a uma desregulação de vias relacionadas a pior prognósticos em LMA. Conclusão: A partir dos dados genômicos, conseguimos observar uma hiperexpressão de CIPC em LMA, levando a suposição de uma regulação negativa do CC, favorece negativamente o quadro de pacientes com LMA. Porém, pesquisas futuras devem se concentrar em elucidar de forma mais clara a expressão e os mecanismos do CIPC na leucemogênese, além de avaliar o seu potencial terapêutico, dado seu papel intrincado na regulação circadiana e a progressão do câncer.

Objective: BCR-ABL1 p190 translocation represents a worse prognosis for acute lymphoblastic leukemia (ALL) patients, and development of new therapeutic approaches is of utmost importance to the oncologic routine. This study aims to evaluate the cytotoxic effect of inhibiting poly-ADP-ribose (PARP) in an ALL Philadelphia positive (Ph+) cell model and validate biomarker differential expression in patient samples. Methodology: Cell lines SUP-B15, Raji and Namalwa were screened for PARP1 expression through qPCR. Non-neoplastic cell line MCR5 was used as a control and ACTB was chosen as the housekeeping gene. Data was compared through one-way ANOVA followed by Bonferroni's post-test. Cells were then treated with either AZD2461, Imatinib or Doxorubicin through serial dilutions and, after 72 hours of treatment, cells were incubated with Alamar Blue and read for fluorescence. Viability results were analyzed through non-linear regression. Finally, samples from 31 p190+ ALL patients were quantified for PARP1 expression and compared to 10 healthy controls. The study was approved by the Ethics Committee of the Federal University of Ceará (5.207.521) and of the Ophir Loyola Hospital (9611320.0.0000.5550), informed written consent was obtained from the patient, being brought into the study only after accepting the terms, and all methods were carried out in accordance with Helsinki guidelines and regulations. Statistical comparison went as previously described for qPCR analysis. All statistical analysis were made using GraphPad Prism (v. 5.01). Results: SUP-B15 considerably overexpresses PARP1 in comparison to both Raji and Namalwa, as well as to the control MRC5 (p < 0.0001), and treatment with AZD2461 showed cytotoxic activity comparable to that of Imatinib in SUP-B15, represented by an IC50 at 72 hours of treatment of 344.3 nM and 329.2 nM, respectively, while showing little to no efficacy when treating either of the remaining cell lines. Validation of biomarker in patient samples reveals an approximate 2-fold increase in PARP1 levels compared to healthy controls (p < 0.01). Discussion: The presence of BCR-ABL1 translocation is highly associated with increased chromosomal instability through intracellular accumulation of reactive oxygen species and promotion of non-conservative repair pathways. The major cytotoxic effect of AZD2461 over SUP-B15, in levels comparable to those of Imatinib, shows that ALL p190+ models seem to be over reliant on pathways that regulate genomic damage and minimize the instability brought by BCR-ABL1, specifically PARP. The overexpression of PARP1 in our experimental model reinforces its translational credibility due to the differential expression of PARP1 detected in p190+ ALL patient samples and sets PARP1 as a potential new target for therapeutic of BCR-ABL1 positive ALL. Conclusion: The use of PARPis shows great effect over the viability of ALL Ph+ models and PARP1 represents a potential new target for investigation in BCR-ABL1 positive tumors. However, studies with larger cohorts of patient samples, as well as in vivo investigations, are still necessary to further elaborate on this hypothesis.

Objetivo: Atualmente, a ampla aplicação da imunofenotipagem e caracterização genética tornou-se fundamental para refinamento diagnóstico, estratificação de risco, predição de prognóstico e promoção de terapia-alvo em pacientes com Leucemia Mieloide Aguda (LMA). Portanto, este estudo visou identificar possíveis correlações entre marcadores imunofenotípicos e estratificações de risco em pacientes com LMA. Metodologia: Esse trabalho obteve aprovação do Comitê de Ética em Pesquisa (nº 4.798.575) e incluiu amostras de sangue periférico e medula óssea de 77 pacientes portadores de LMA atendidos no Hospital Geral de Fortaleza (HGF). Todos os pacientes foram submetidos a testes de imunofenotipagem, cariótipo e biologia molecular para que posteriormente fossem estratificados seguindo as diretrizes estabelecidas pelo European Leukemia Net de 2022. Os pacientes com foram submetidos à pesquisa das seguintes alterações: FLT3-ITD, RUNX1::RUNX1T1, CBFB::MYH11 e PML::RARA através da técnica de RT-qPCR. Resultados: Entre os 77 pacientes com LMA, 8 foram classificados com risco favorável, 54 com risco intermediário e 15 com risco adverso. Os fenótipos mais frequentes com o grupo de risco favorável foram CD45, CD33, CD34, CD117, HLA-DR e MPO. Em relação ao grupo de risco intermediário, os marcadores mais incidentes foram CD33, CD45, CD117, CD13, CD34 e HLA-DR. Por sua vez, os principais fenótipos associados ao grupo de risco adverso foram CD117, CD34, CD33, CD45 e CD13. Em geral, foi observada a associação de dos marcadores CD7 e CD10 com maiores taxas de óbito. Em relação às alterações genéticas que conferem um risco favorável, 1 paciente possuía a translocação CBFB::MYH11 e7 apresentavam a translocação RUNX1::RUNX1T1. Quanto às alterações genéticas associadas ao risco intermediário, 4 pacientes apresentavam a mutação FLT3-ITD e 10 possuíam a translocação PML::RARA. Discussão: Estudos reportaram que taxas de sobrevida global e de remissão completa aparentam ser influenciadas pela expressão de determinados marcadores, como CD10, CD13, CD34, CD41, CD61, CD117 e HLA-DR. Estudos relataram maior expressão de CD33, CD34, CD117, HLA-DR, CD15, CD64 e CD14 em paciente com a fusão CBFB::MYH11, enquanto este trabalho observou expressão de CD45, CD34, CD227, MPO, CD38, HLA-DR e CD33. Os imunofenótipos dos pacientes com RUNX1::RUNX1T1 identificados neste estudo corroboraram com os marcadores descritos na literatura: CD34, CD117, CD13, CD33, CD19, CD56, CD38, HLA-DR e MPO. Outros trabalhos ainda identificaram fenótipos que aparentam estar associados a alterações genéticas, como os fenótipos CD33, CD34, CD123, CD25 e CD99 que predizem FLT3-ITD. Entretanto, no presente estudo foram observados CD64, CD45, CD33 e HLA-DR associados à mutação FLT3-ITD. Os pacientes com PML::RARA analisados neste estudo apresentaram os marcadores CD33, CD64, CD117, CD13 e MPO, corroborando com outros trabalhos na literatura. Conclusão: Através deste estudo foi possível destacar a existência de correlação entre a expressão de determinados imunofenótipos e a estratificação de risco de pacientes com LMA. A identificação dessas informações pode proporcionar um diagnóstico mais rápido e preciso, além de estratégias de tratamento mais bem direcionadas.

Objetivo: Diante da importância da detecção de alterações genéticas em pacientes com leucemias, o objetivo desse estudo foi comparar a sensibilidade das técnicas Nested -PCR e RT-qPCR na detecção de alterações genéticas em pacientes portadores de leucemias agudas. Metodologia: Aprovado pelo Comitê de Ética em Pesquisa (nº4.798.575), este estudo incluiu amostras de sangue periférico e medula óssea de 117 pacientes atendidos no Hospital Geral de Fortaleza (HGF). Após a obtenção dos respectivos diagnósticos de leucemia mieloide aguda e leucemia linfoblástica aguda, todas as amostras foram submetidas a análises qualitativas através da técnica de Nested -PCR e de expressão quantitativa através da técnica de RT-qPCR. Os pacientes com LMA foram submetidos à análise das seguintes alterações: FLT3-ITD, RUNX1::RUNX1T1, CBFB::MYH11 e PML::RARA, enquanto foram pesquisadas as fusões BCR::ABL1, TCF3::PBX1, KMT2A::AFF1, ETV6::RUNX1 e SIL::TAL1 nas amostras dos pacientes com LLA. Resultados: Ao longo do estudo, foram diagnosticados 77 pacientes com Leucemia Mieloide Aguda (LMA) e 40 com Leucemia Linfoblástica Aguda (LLA), dos quais 35 correspondiam ao tipo B e 5 ao tipo T. Entre os 77 pacientes com LMA, foram detectados 4 pacientes com a mutação FLT3-ITD, 7 pacientes com a translocação RUNX1::RUNX1T1, 10 pacientes com a translocação PML::RARA e 1 paciente com a translocação CBFB::MYH11. A presença dessas alterações não foi detectada através da técnica Nested -PCR, porém foi possível realizar essa identificação através do método de RT-qPCR. Já entre os 40 pacientes portadores de LLA, observou-se a presença de 24 pacientes com a translocação BCR::ABL1 e 3 pacientes com a translocação TCF3::PBX1, através da metodologia RT-qPCR. Diferentemente das análises realizadas nas amostras de LMA, foi possível identificar a formação de bandas na pesquisa pelas fusões nas amostras de LLA. Entretanto, a alteração TCF3::PBX1 foi detectada pela Nested -PCR apenas em amostras de medula óssea dos pacientes, mesmo apresentando alta leucometria e taxas acima de 20% de blastos circulantes em sangue periférico. Discussão: O presente estudo demonstrou que a técnica de RT-qPCR apresentou uma sensibilidade maior em comparação à técnica de Nested -PCR no momento do diagnóstico das amostras de leucemias agudas estudadas. Isso também foi visto em outros trabalhos que demonstraram que a metodologia de RT-qPCR é mais sensível e rápida na detecção de diversas doenças quando comparada à Nested -PCR. A literatura traz resultados controversos em relação à Nested -PCR, já que alguns autores determinaram essa metodologia como um método confiável e com alta sensibilidade para o diagnóstico de diversas doenças, enquanto outros pesquisadores indicam a utilização de métodos mais eficientes como a RT-qPCR. No geral, a utilização da técnica Nested-PCR aparenta ser mais indicada em casos que o diagnóstico por PCR convencional simples não é o suficiente. Conclusão: Portanto, o estudo demonstrou que a técnica de RT-qPCR apresentou uma sensibilidade maior em comparação à técnica de Nested -PCR no momento do diagnóstico das amostras de leucemias agudas estudadas. A técnica de RT-qPCR se mostrou muito eficiente para o diagnóstico rápido e sensível de alterações genéticas em ambos os tipos de amostras analisadas no estudo, caracterizando uma grande vantagem, já que deixa de exigir coletas invasivas de medula óssea.

Introduction: Global burden of stroke mortality tended to be higher among men. A disproportionate stroke burden has been placed upon women, primarily because of life expectancy, putting a more significant burden in this population. The purpose of this study was to address sex differences in stroke epidemiology and treatment in Fortaleza, Brazil., Methods: Between April 2009 and 2012, data were prospectively gathered from 19 hospitals, following the WHO's Stroke Steps program, stratified by sex assigned at birth., Results: We included 4,679 patients, 2,403 females. Women mean age was 69.1 (±15.2) years and 66.2 (±13.5) years in males (p < 0.01). Females were more often white, while time to hospital admission and CT scan from symptom onset were similar across sexes. Men more commonly presented with motor, speech, and sensory symptoms, whereas women were more likely to present with reduced consciousness and headaches. Additionally, women had higher rates of diabetes and obesity. Men were more frequently smokers and had higher rates of alcohol misuse, as well as a history of heart attacks or strokes. We utilized univariable logistic regression to assess mRS scores at discharge, in scores 0-2 (lesser disability) and scores 3-6 (more significant disability). The dependent variable regards scores that are above 3. Fewer women than men achieved a modified Rankin Score of ≤2 (77.6% versus 81.7%; p < 0.01). Multivariable analysis identified the female gender as an independent predictor of having a higher mRS score at discharge (OR 1.23; 95% CI [1.01-1.51]; p = 0.04)., Conclusion: Our study, one of the largest South American epidemiological studies on patients admitted with stroke, highlights the sex-specific nuances in stroke outcomes. Our findings underscore that risk factors for stroke vary significantly between men and women, thereby necessitating tailored preventive strategies., (© 2024 S. Karger AG, Basel.)

Background/aim: Although the reciprocal translocation t(9;22)(q34;q11) is a hallmark of chronic myeloid leukemia (CML), it is also present in acute lymphoblastic leukemia (ALL) and acute myeloid leukemia (AML). Depending on the gene's breakpoint, it is possible to obtain three isoforms, among which p190 stands out for the poor prognosis it induces whenever it appears. Due to the genomic instability induced by BCR::ABL1, it is proposed to expand the applicability of poly-ADP-ribose polymerase-1 (PARP1) and its inhibitors in hematological neoplasms., Materials and Methods: We measured the expression levels of PARP1 by quantitative real-time PCR (qPCR) using TaqMan®, correlating its expression with BCR::ABL1 p190+, to evaluate its influence in the clinic of adult patients., Results: We found that PARP1 is expressed differently in ALL, AML and CML and that p190 transcripts do not follow a linear pattern in these populations. We also found that PARP1 expression is not correlated with age, white blood cell and the amount of p190 transcripts., Conclusion: Despite the lack of statistical correlation between the variables analyzed, the role of PARP1 in BCR::ABL1 leukemia cannot be ruled out, given the instability profile promoted by this translocation. Finally, further studies involving a larger sample of patients are needed, as well as investigations into other molecular pathways that may impact on the pathogenesis of different BCR::ABL1 leukemic subtypes., Competing Interests: The Authors declare no conflicts of interest regarding this study., (©2024 The Author(s). Published by the International Institute of Anticancer Research.)

Background/aim: Acute myeloid leukemia (AML) is a myeloproliferative neoplasm marked by abnormal clonal expansion of hematopoietic progenitor cells, displaying karyotypic aberrations and genetic mutations as prognostic indicators. The World Health Organization (WHO) and the European LeukemiaNet guidelines categorize BCR::ABL1 p190+ AML as high risk. This study explored the identification of the increased incidence of BCR::ABL1 p190+ in our AML population., Patients and Methods: This study included 96 AML patients stratified according to WHO guidelines. Subsequently, patients were screened for genetic abnormalities, such as BCR::ABL1 p 190+, PML::RARA, RUNX1::RUNX1T1, and CBFB::MYH11 by quantitative reverse transcription polymerase chain reaction (RT-qPCR) analysis., Results: Among 96 AML patients, 36 displayed BCR::ABL1 p190+, overcoming the expected global incidence. Age variations (19 to 78 years) showed no significant laboratory differences between BCR::ABL1 p190+ and non-BCR::ABL p190+ cases. The overall survival analysis revealed no statistically significant differences among the patients (p=0.786)., Conclusion: The analyzed population presented a higher frequency of BCR::ABL1 p190+ detection in adult AML patients when compared to what is described in the worldwide literature. Therefore, more studies are needed to establish the reason why this incidence is higher and what the best treatment approach should be in these cases., (Copyright © 2024, International Institute of Anticancer Research (Dr. George J. Delinasios), All rights reserved.)

The year 2021 marked a decade of holopelagic sargassum (morphotypes Sargassum natans I and VIII, and Sargassum fluitans III) stranding on the Caribbean and West African coasts. Beaching of millions of tons of sargassum negatively impacts coastal ecosystems, economies, and human health. Additionally, the La Soufrière volcano erupted in St. Vincent in April 2021, at the start of the sargassum season. We investigated potential monthly variations in morphotype abundance and biomass composition of sargassum harvested in Jamaica and assessed the influence of processing methods (shade-drying vs. frozen samples) and of volcanic ash exposure on biochemical and elemental components. S. fluitans III was the most abundant morphotype across the year. Limited monthly variations were observed for key brown algal components (phlorotannins, fucoxanthin, and alginate). Shade-drying did not significantly alter the contents of proteins but affected levels of phlorotannins, fucoxanthin, mannitol, and alginate. Simulation of sargassum and volcanic ash drift combined with age statistics suggested that sargassum potentially shared the surface layer with ash for ~50 d, approximately 100 d before stranding in Jamaica. Integrated elemental analysis of volcanic ash, ambient seawater, and sargassum biomass showed that algae harvested from August had accumulated P, Al, Fe, Mn, Zn, and Ni, probably from the ash, and contained less As. This ash fingerprint confirmed the geographical origin and drift timescale of sargassum. Since environmental conditions and processing methods influence biomass composition, efforts should continue to improve understanding, forecasting, monitoring, and valorizing sargassum, particularly as strandings of sargassum show no sign of abating., Competing Interests: Competing interests statement:The authors declare no competing interest.

Background/aim: The relevance of cytogenetic markers as prognostic risk factors has been demonstrated in a vast number of studies, with many prognostication tools utilizing these factors to determine treatment approaches. Patients aged above 60 years represent an important subgroup of acute myeloid leukemia (AML) patients, especially because they usually exhibit a poorer cytogenetic landscape and are less suitable for intensive treatments. The importance of evaluating prognostic parameters in AML, especially in low-income countries, prompted an investigation into CD38 expression and its effects., Patients and Methods: Medical records of AML patients aged above 60 years from three hospitals in Brazil's northwest region were analyzed. A total of 67 patients were evaluated in terms of overall survival and factors predicting worse outcomes. The risk stratification was performed based on the European LeukemiaNet 2022 guidelines. The analysis of immunophenotyping markers was conducted using multi-parametric flow cytometry., Results: The overall survival of CD38-positive AML patients was higher than that of patients with CD38-negative AML, with survival rates of 15.6 months versus 4 months, respectively (p-value=0.026). The impact of CD38 positivity was relevant also in multivariable Cox proportional hazards regression, demonstrating a positive effect on overall survival, with a hazard ratio of 0.33 (95%CI=0.13-0.79; p-value=0.014)., Conclusion: Expression of CD38 in patients with AML was associated with better overall survival and serves as a relevant predictor of improved outcome in patients aged above 60 years., (Copyright © 2024 International Institute of Anticancer Research (Dr. George J. Delinasios), All rights reserved.)

This article centres on contemporary mergers between state security practices, popular media and religion in Rio de Janeiro, Brazil. It discusses the actions and the representations of BOPE - Batalhão de Operações Especiais da Polícia Militar do Rio de Janeiro - the special operations battalion of the military police of Rio de Janeiro, to show how this unit frequently employs Christian symbols and practices to legitimate violence at the border between the legal and the illegal. BOPE's religious iconography is amplified by movies, memes, toys and popular clothing. BOPE's military pop-culture can best be regarded as theopolitical because such a perspective underscores the fact that different Christian symbols and practices, derived from Roman Catholicism and Pentecostalism, produce incoherent political theologies that are fuelled by bottom-up innovations and institutional legacies., Competing Interests: No potential conflict of interest was reported by the author(s)., (© 2024 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group.)

Leukemias are hematological neoplasms characterized by dysregulations in several cellular signaling pathways, prominently including the PI3K/AKT/mTOR pathway. Since this pathway is associated with several important cellular mechanisms, such as proliferation, metabolism, survival, and cell death, its hyperactivation significantly contributes to the development of leukemias. In addition, it is a crucial prognostic factor, often correlated with therapeutic resistance. Changes in the PI3K/AKT/mTOR pathway are identified in more than 50% of cases of acute leukemia, especially in myeloid lineages. Furthermore, these changes are highly frequent in cases of chronic lymphocytic leukemia, especially those with a B cell phenotype, due to the correlation between the hyperactivation of B cell receptors and the abnormal activation of PI3Kδ. Thus, the search for new therapies that inhibit the activity of the PI3K/AKT/mTOR pathway has become the objective of several clinical studies that aim to replace conventional oncological treatments that have high rates of toxicities and low specificity with target-specific therapies offering improved patient quality of life. In this review we describe the PI3K/AKT/mTOR signal transduction pathway and its implications in leukemogenesis. Furthermore, we provide an overview of clinical trials that employed PI3K/AKT/mTOR inhibitors either as monotherapy or in combination with other cytotoxic agents for treating patients with various types of leukemias. The varying degrees of treatment efficacy are also reported., Competing Interests: The Authors declare no conflict of interest in relation to this study. The funders had no role in the design of the study; in the collection, analyses, or data interpretation; in the writing of the manuscript, or in the decision to publish the results., (Copyright 2024, International Institute of Anticancer Research.)

Dysregulated neuronal excitability is a hallmark of amyotrophic lateral sclerosis (ALS). We sought to investigate how functional changes to the axon initial segment (AIS), the site of action potential generation, could impact neuronal excitability in ALS human induced pluripotent stem cell (hiPSC) motor neurons. We find that early TDP-43 and C9orf72 hiPSC motor neurons show an increase in the length of the AIS and impaired activity-dependent AIS plasticity that is linked to abnormal homeostatic regulation of neuronal activity and intrinsic hyperexcitability. In turn, these hyperactive neurons drive increased spontaneous myofiber contractions of in vitro hiPSC motor units. In contrast, late hiPSC and postmortem ALS motor neurons show AIS shortening, and hiPSC motor neurons progress to hypoexcitability. At a molecular level, aberrant expression of the AIS master scaffolding protein ankyrin-G and AIS-specific voltage-gated sodium channels mirror these dynamic changes in AIS function and excitability. Our results point toward the AIS as an important site of dysfunction in ALS motor neurons., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.)

Detection of t(9;22), and consequent BCR::ABL1 fusion, is still a marker of worse prognosis for acute lymphoblastic leukemia (ALL), with resistance to tyrosine-kinase inhibitor therapy being a major obstacle in the clinical practice for this subset of patients. In this study, we investigated the effectiveness of targeting poly-ADP-ribose polymerase (PARP) in a model of BCR::ABL1 p190+ ALL, the most common isoform to afflict ALL patients, and demonstrated the use of experimental PARP inhibitor (PARPi), AZD2461, as a therapeutic option with cytotoxic capabilities similar to that of imatinib, the current gold standard in medical care. We characterized cytostatic profiles, induced cell death, and biomarker expression modulation utilizing cell models, also providing a comprehensive genome-wide analysis through an aCGH of the model used, and further validated PARP1 differential expression in samples of ALL p190+ patients from local healthcare institutions, as well as in larger cohorts of online and readily available datasets. Overall, we demonstrate the effectiveness of PARPi in the treatment of BCR::ABL1 p190+ ALL cell models and that PARP1 is differentially expressed in patient samples. We hope our findings help expand the characterization of molecular profiles in ALL settings and guide future investigations into novel biomarker detection and pharmacological choices in clinical practice.

Past sea level changes and geological instability along watershed boundaries have largely influenced fish distribution across coastal basins, either by dispersal via palaeodrainages now submerged or by headwater captures, respectively. Accordingly, the South American Atlantic coast encompasses several small and isolated drainages that share a similar species composition, representing a suitable model to infer historical processes. Leporinus bahiensis is a freshwater fish species widespread along adjacent coastal basins over narrow continental shelf with no evidence of palaeodrainage connections at low sea level periods. Therefore, this study aimed to reconstruct its evolutionary history to infer the role of headwater captures in the dispersal process. To accomplish this, we employed molecular-level phylogenetic and population structure analyses based on Sanger sequences (5 genes) and genome-wide SNP data. Phylogenetic trees based on Sanger data were inconclusive, but SNPs data did support the monophyletic status of L. bahiensis. Both COI and SNP data revealed structured populations according to each hydrographic basin. Species delimitation analyses revealed from 3 (COI) to 5 (multilocus approach) MOTUs, corresponding to the sampled basins. An intricate biogeographic scenario was inferred and supported by Approximate Bayesian Computation (ABC) analysis. Specifically, a staggered pattern was revealed and characterized by sequential headwater captures from basins adjacent to upland drainages into small coastal basins at different periods. These headwater captures resulted in dispersal throughout contiguous coastal basins, followed by deep genetic divergence among lineages. To decipher such recent divergences, as herein represented by L. bahiensis populations, we used genome-wide SNPs data. Indeed, the combined use of genome-wide SNPs data and ABC method allowed us to reconstruct the evolutionary history and speciation of L. bahiensis. This framework might be useful in disentangling the diversification process in other neotropical fishes subject to a reticulate geological history., (© The Author(s) 2023. Published by Oxford University Press on behalf of the Society of Systematic Biologists.)

Bardet-Biedl syndrome (BBS) is a ciliopathy with pleiotropic effects on multiple tissues, including the kidney. Here we have compared renal differentiation of iPS cells from healthy and BBS donors. High content image analysis of WT1-expressing kidney progenitors showed that cell proliferation, differentiation and cell shape were similar in healthy, BBS1 , BBS2 , and BBS10 mutant lines. We then examined three patient lines with BBS10 mutations in a 3D kidney organoid system. The line with the most deleterious mutation, with low BBS10 expression, expressed kidney marker genes but failed to generate 3D organoids. The other two patient lines expressed near normal levels of BBS10 mRNA and generated multiple kidney lineages within organoids when examined at day 20 of organoid differentiation. However, on prolonged culture (day 27) the proximal tubule compartment degenerated. Introducing wild type BBS10 into the most severely affected patient line restored organoid formation, whereas CRISPR-mediated generation of a truncating BBS10 mutation in a healthy line resulted in failure to generate organoids. Our findings provide a basis for further mechanistic studies of the role of BBS10 in the kidney., 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 Williams, Hurling, Munir, Harley, Machado, Cujba, Alvarez-Fallas, Danovi, Lieberam, Sancho, Beales and Watt.)

Surubim (Pseudoplatystoma corruscans, Pimelodidae) are migratory catfish native to the rivers in the La Plata and São Francisco basins. They are piscivores that attain considerable body sizes and are a valuable economic resource. Surubim exhibits extensive migrations during its life cycle that may affect the population structure at vast geographic scales. The authors examined the genetic diversity and population genetic structure of P. corruscans using microsatellite markers from a comprehensive sampling of 260 individuals from the Upper and Lower Paraná River. They identified two well-differentiated genetic clusters corresponding to a natural geographic barrier historically separating Upper and Lower Paraná regions. They also demonstrated temporal variation in population genetic structure at a site in Lower Paraná close to the confluence with the Paraguay River, most likely explained by the influx of migrant fishes at certain times of the year., (© 2023 Fisheries Society of the British Isles.)

Acute myeloid leukemia (AML) is a hematologic malignancy that occurs due to alterations such as genetic mutations, chromosomal translocations, or changes in molecular levels. These alterations can accumulate in stem cells and hematopoietic progenitors, leading to the development of AML, which has a prevalence of 80% of acute leukemias in the adult population. Recurrent cytogenetic abnormalities, in addition to mediating leukemogenesis onset, participate in its evolution and can be used as established diagnostic and prognostic markers. Most of these mutations confer resistance to the traditionally used treatments and, therefore, the aberrant protein products are also considered therapeutic targets. The surface antigens of a cell are characterized through immunophenotyping, which has the ability to identify and differentiate the degrees of maturation and the lineage of the target cell, whether benign or malignant. With this, we seek to establish a relationship according to the molecular aberrations and immunophenotypic alterations that cells with AML present.

V3 spinal interneurons are a key element of the spinal circuits, which control motor function. However, to date, there are no effective ways of deriving a pure V3 population from human pluripotent stem cells. Here, we report a method for differentiation and isolation of spinal V3 interneurons, combining extrinsic factor-mediated differentiation and magnetic activated cell sorting. We found that differentiation of V3 progenitors can be enhanced with a higher concentration of Sonic Hedgehog agonist, as well as culturing cells in 3D format. To enable V3 progenitor purification from mixed differentiation cultures, we developed a transgene reporter, with a part of the regulatory region of V3-specific gene Nkx2-2 driving the expression of a membrane marker CD14. We found that in human cells, NKX2-2 initially exhibited co-labelling with motor neuron progenitor marker, but V3 specificity emerged as the differentiation culture progressed. At these later differentiation timepoints, we were able to enrich V3 progenitors labelled with CD14 to ~ 95% purity, and mature them to postmitotic V3 interneurons. This purification tool for V3 interneurons will be useful for in vitro disease modeling, studies of normal human neural development and potential cell therapies for disorders of the spinal cord., (© 2023. The Author(s).)

The purpose of this study was to compare the effects of myofascial release and muscle energy on acute outcomes in trunk extensors active range-of-motion and strength in recreationally resistance-trained women. Seventeen apparently healthy women performed three experimental protocols using a cross-over, randomized (counterbalanced in Latin Square format), and within-subjects design: a) range-of-motion and strength test after a manual myofascial release protocol (MFR); b) flexibility and strength test after a muscle energy protocol (ME); and c) range-of-motion and strength test without myofascial release or muscle energy (control condition). Active trunk range-of-motion was measured via a sit-and-reach test and trunk extension strength via isometric dorsal dynamometer. A significant increase in range-of-motion was found for MFR (p = 0.002; d = 0.71) and ME (p < 0.001; d = 0.47) when comparing post-intervention with baseline values. Similarly, a significant increase for strength was found for MFR (p = 0.018; d = 0.10) when comparing post-intervention with baseline values. In conclusion, both techniques (MFR and ME) improved trunk range-of-motion with the sit and reach test immediately post-intervention; however, MFR showed greater magnitude increases in range-of-motion (MFR: (medium magnitude) vs ME: small magnitude). Due to the potential health implications, both (MFR and ME) responses should be among the many considerations for rehabilitation and performance exercise prescription when prescribing an exercise regimen., Competing Interests: Declaration of competing interest None., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

The WD repeat containing antisense to TP53 ( WRAP53 ) gene codifies an antisense transcript for tumor protein p53 ( TP53 ), stabilization (WRAP53α), and a functional protein (WRAP53β, WDR79, or TCAB1). The WRAP53β protein functions as a scaffolding protein that is important for telomerase localization, telomere assembly, Cajal body integrity, and DNA double-strand break repair. WRAP53β is one of many proteins known for containing WD40 domains, which are responsible for mediating a variety of cell interactions. Currently, WRAP53 overexpression is considered a biomarker for a diverse subset of cancer types, and in this study, we describe what is known about WRAP53β's multiple interactions in cell protein trafficking, Cajal body formation, and DNA double-strand break repair and its current perspectives as a biomarker for cancer.

Multiple myeloma (MM) is a blood cell neoplasm characterized by excessive production of malignant monoclonal plasma cells (activated B lymphocytes) by the bone marrow, which end up synthesizing antibodies or antibody fragments, called M proteins, in excess. The accumulation of this production, both cells themselves and of the immunoglobulins, causes a series of problems for the patient, of a systemic and local nature, such as blood hyperviscosity, renal failure, anemia, bone lesions, and infections due to compromised immunity. MM is the third most common hematological neoplasm, constituting 1% of all cancer cases, and is a disease that is difficult to treat, still being considered an incurable disease. The treatments currently available cannot cure the patient, but only extend their lifespan, and the main and most effective alternative is autologous hematopoietic stem cell transplantation, but not every patient is eligible, often due to age and pre-existing comorbidities. In this context, the search for new therapies that can bring better results to patients is of utmost importance. Protein tyrosine kinases (PTKs) are involved in several biological processes, such as cell growth regulation and proliferation, thus, mutations that affect their functionality can have a great impact on crucial molecular pathways in the cells, leading to tumorigenesis. In the past couple of decades, the use of small-molecule inhibitors, which include tyrosine kinase inhibitors (TKIs), has been a hallmark in the treatment of hematological malignancies, and MM patients may also benefit from TKI-based treatment strategies. In this review, we seek to understand the applicability of TKIs used in MM clinical trials in the last 10 years.

The circadian clock (CC) is a daily system that regulates the oscillations of physiological processes and can respond to the external environment in order to maintain internal homeostasis. For the functioning of the CC, the clock genes (CG) act in different metabolic pathways through the clock-controlled genes (CCG), providing cellular regulation. The CC's interruption can result in the development of different diseases, such as neurodegenerative and metabolic disorders, as well as cancer. Leukemias correspond to a group of malignancies of the blood and bone marrow that occur when alterations in normal cellular regulatory processes cause the uncontrolled proliferation of hematopoietic stem cells. This review aimed to associate a deregulated CC with the manifestation of leukemia, looking for possible pathways involving CG and their possible role as leukemic biomarkers.

Hematopoietic stem cells (HSCs) are known for their ability to proliferate and self-renew, thus being responsible for sustaining the hematopoietic system and residing in the bone marrow (BM). Leukemic stem cells (LSCs) are recognized by their stemness features such as drug resistance, self-renewal, and undifferentiated state. LSCs are also present in BM, being found in only 0.1%, approximately. This makes their identification and even their differentiation difficult since, despite the mutations, they are cells that still have many similarities with HSCs. Although the common characteristics, LSCs are heterogeneous cells and have different phenotypic characteristics, genetic mutations, and metabolic alterations. This whole set of alterations enables the cell to initiate the process of carcinogenesis, in addition to conferring drug resistance and providing relapses. The study of LSCs has been evolving and its application can help patients, where through its count as a biomarker, it can indicate a prognostic factor and reveal treatment results. The selection of a target to LSC therapy is fundamental. Ideally, the target chosen should be highly expressed by LSCs, highly selective, absence of expression on other cells, in particular HSC, and preferentially expressed by high numbers of patients. In view of the large number of similarities between LSCs and HSCs, it is not surprising that current treatment approaches are limited. In this mini review we seek to describe the immunophenotypic characteristics and mechanisms of resistance presented by LSCs, also approaching possible alternatives for the treatment of patients., 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 © 2022 Barreto, Pessoa, Machado, Pantoja, Ribeiro, Lopes, Amaral de Moraes, de Moraes Filho, de Souza, Burbano, Khayat and Moreira-Nunes.)

Human T cell leukemia virus type 1 (HTLV-1) was identified as the first pathogenic human retrovirus and is estimated to infect 5 to 10 million individuals worldwide. Unlike other retroviruses, there is no effective therapy to prevent the onset of the most alarming diseases caused by HTLV-1, and the more severe cases manifest as the malignant phenotype of adult T cell leukemia (ATL). MicroRNA (miRNA) dysfunction is a common feature of leukemogenesis, and it is no different in ATL cases. Therefore, we sought to analyze studies that reported deregulated miRNA expression in HTLV-1 infected cells and patients' samples to understand how this deregulation could induce malignancy. Through in silico analysis, we identified 12 miRNAs that stood out in the prediction of targets, and we performed functional annotation of the genes linked to these 12 miRNAs that appeared to have a major biological interaction. A total of 90 genes were enriched in 14 KEGG pathways with significant values, including TP53, WNT, MAPK, TGF-β, and Ras signaling pathways. These miRNAs and gene interactions are discussed in further detail for elucidation of how they may act as probable drivers for ATL onset, and while our data provide solid starting points for comprehension of miRNAs' roles in HTLV-1 infection, continuous effort in oncologic research is still needed to improve our understanding of HTLV-1 induced leukemia.

Assessing the relative contribution of Land Use and Cover Changes (LUCC) and climate changes on runoff still represents a great challenge for water resources management. This issue is particularly critical for the Upper Paraná River Basin (UPRB), one of the most important basins in South America and responsible for most of the production of food, ethanol, and electricity generation in Brazil. In this paper, we used the Soil and Water Assessment Tool (SWAT) to quantitatively assess the relative contribution of both forcings. The simulation period included a time of great importance for climate studies, known as the 1970s global climate shift, and of great impact on river discharge within the UPRB. Three land use and cover scenarios were assigned to the 1961-1990 period of simulations, representing land use and cover during a pristine period (around the Year 1500), 1960, and 1985. Thirteen years of precipitation before and after the climate shift (considered to be the period 1974-1977) were analyzed and compared. Results showed a precipitation increase for the basin in general after the climate shift. The increase in rainfall reached up to 15% in many northern areas and more than 20% in the southern parts of the basin. By comparing all simulations, results indicate that both LUCC and precipitation increase due to the climate shift had a significant effect on the changes in annual discharge of the largest rivers of the UPRB. However, the results suggest that the impact of the precipitation increase on the discharge exceeded that of the LUCC. Between 1960 and 1985 the LUCC accounts for about 16% of the increase of the median annual discharge, whereas climate shift accounts for an increase of about 32%. These findings, suggesting a more relevant role for the climate, are consistent with two recent water crisis experienced by the country in the last decades, caused by prolonged below-normal rainfall throughout 2001/2002 and again in 2014/2015., (Copyright © 2022 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Pelagic Sargassum species have been known for centuries in the Sargasso Sea of the North Atlantic Ocean. In 2011, a new area concentrating high biomass of these brown algae started developing in the Tropical Atlantic Ocean. Since then, massive and recurrent Sargassum influxes have been reported in the Caribbean and off the coast of Western Africa. These Sargassum events have a major negative impact on coastal ecosystems and nearshore marine life, and affect socio-economic sectors, including public health, coastal living, tourism, fisheries, and maritime transport. Despite recent advances in the forecasting of Sargassum events, and elucidation of the seaweed composition, many knowledge gaps remain, including morphotype abundance during Sargassum events, drift of the seaweeds in the months prior to stranding, and influence of sample processing methods on biomass biochemical composition. Using seaweeds harvested on the coasts of Jamaica in summer of 2020, we observed that S. fluitans III was the most abundant morphotype at different times and sampling locations. No clear difference in the geographical origin, or provenance, of the Sargassum mats was observed. The majority of Sargassum backtracked from both north and south of Jamaica experienced ambient temperatures of around 27 °C and salinity in the range of 34-36 psu before stranding. We also showed that cheap (sun) compared to expensive (freeze) drying techniques influence the biochemical composition of biomass. Sun-drying increased the proportion of phenolic compounds, but had a deleterious impact on fucoxanthin content and on the quantities of monosaccharides, except for mannitol. Effects on the content of fucose containing sulfated polysaccharides depended on the method used for their extraction, and limited variation was observed in ash, protein, and fatty acid content within most of the sample locations investigated. These observations are important for the storage and transport of the biomass in the context of its valorisation., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 The Authors. Published by Elsevier B.V. All rights reserved.)

The increasing numbers of cancer cases worldwide and the exceedingly high mortality rates of some tumor subtypes raise the question about if the current protocols for cancer management are effective and what has been done to improve upon oncologic patients' prognoses. The traditional chemo-immunotherapy options for cancer treatment focus on the use of cytotoxic agents that are able to overcome neoplastic clones' survival mechanisms and induce apoptosis, as well as on the ability to capacitate the host's immune system to hinder the continuous growth of malignant cells. The need to avert the highly toxic profiles of conventional chemo-immunotherapy and to overcome the emerging cases of tumor multidrug resistance has fueled a growing interest in the field of precision medicine and targeted molecular therapies in the last couple of decades, although relatively new alternatives in oncologic practices, the increased specificity, and the positive clinical outcomes achieved through targeted molecular therapies have already consolidated them as promising prospects for the future of cancer management. In recent years, the development and application of targeted drugs as tyrosine kinase inhibitors have enabled cancer treatment to enter the era of specificity. In addition, the combined use of targeted therapy, immunotherapy, and traditional chemotherapy has innovated the standard treatment for many malignancies, bringing new light to patients with recurrent tumors. This article comprises a series of clinical trials that, in the past 5 years, utilized kinase inhibitors (KIs) as a monotherapy or in combination with other cytotoxic agents to treat patients afflicted with solid tumors. The results, with varying degrees of efficacy, are reported.

India ranks the second in the world in terms of its sheep population with approximately 74.26 million represented by 44 well-described breeds in addition to several non-descript populations. Genetic diversity and phylogeography of Indian sheep breeds remain poorly understood, particularly for south Indian breeds. To have a comprehensive view of the domestication history of Indian sheep, we sequenced the mitochondrial DNA (mtDNA) control region (D-loop) and cytochrome b gene (CYTB) of 16 Indian domestic sheep breeds, most of them (13) from the south India. We analysed these sequences along with published data of domestic and wild sheep from different countries, including India. The haplotype diversity was relatively high in Indian sheep, which were classified into the three known mtDNA lineages, namely A, B and C. Lineage A was predominant among Indian sheep whereas lineages B and C were observed at low frequencies but C was restricted to the breeds of north and east India. The median joining network showed five major expanding haplogroups of lineage A (A1-A5). Out of which, A2, A4 and A5 were more frequent in Indian sheep in contrast to breeds from other parts of the world. Among the 27 Indian sheep breeds analysed, Mandya and Sonadi breeds were significantly different from other Indian breeds in the MDS analyses. This was explained by a very high contribution of lineage B into these two breeds. The Approximate Bayesian Computation (ABC) provided evidence for the domestication of lineage A sheep in the Indian subcontinent. Contrary to the current knowledge, we also found strong support for the introduction of lineage B into Indian subcontinent through sea route rather than from the Mongolian Plateau. The neighbour-joining tree of domestic and wild sheep revealed the close genetic relationship of Indian domestic sheep with Pakistani wild sheep O. vignei blanfordi. Based on our analyses and archaeological evidences, we suggest the Indian subcontinent as one of the domestication centres of the lineage A sheep, while lineage B sheep might have arrived into India from elsewhere via Arabian sea route. To the best of our knowledge, this is the first comprehensive study on Indian sheep where we have analysed more than 740 animals belonging to 27 sheep breeds raised in various regions of India. Our study provides insight into the understanding of the origin and migratory history of Indian sheep., (© 2021. The Author(s).)

Cancer is still a major barrier to life expectancy increase worldwide, and hematologic neoplasms represent a relevant percentage of cancer incidence rates. Tumor dependence of continuous proliferative signals mediated through protein kinases overexpression instigated increased strategies of kinase inhibition in the oncologic practice over the last couple decades, and in this review, we focused our discussion on relevant clinical trials of the past five years that investigated kinase inhibitor (KI) usage in patients afflicted with relapsed/refractory (R/R) hematologic malignancies as well as in the pharmacological characteristics of available KIs and the dissertation about traditional chemotherapy treatment approaches and its hindrances. A trend towards investigations on KI usage for the treatment of chronic lymphoid leukemia and acute myeloid leukemia in R/R settings was observed, and it likely reflects the existence of already established treatment protocols for chronic myeloid leukemia and acute lymphoid leukemia patient cohorts. Overall, regimens of KI treatment are clinically manageable, and results are especially effective when allied with tumor genetic profiles, giving rise to encouraging future prospects of an era where chemotherapy-free treatment regimens are a reality for many oncologic patients.

A purified exo-polygalacturonase of Neosartorya glabra (EplNg) was successfully characterized. EplNg native presented 68.2 kDa, with 32% carbohydrate content. The deglycosylated form showed 46.3 kDa and isoelectric point of 5.4. The identity of EplNg was confirmed as an exo-polygalacturonase class I (EC 3.2.1.67) using mass spectrometry and Western-Blotting. Capillary electrophoresis indicated that only galacturonic acid was released by the action of EplNg on sodium polypectate, confirming an exoenzyme character. The structural model confers that EplNg has a core formed by twisted parallel β-sheets structure. Among twelve putative cysteines, ten were predicted to form disulfide bridges. The catalytic triad predicted is composed of Asp 223 , Asp 245 , and Asp 246 aligned along with a distance in 4-5 Å, suggesting that EplNg probably does not perform the standard inverting catalytic mechanism described for the GH28 family. EplNg was active from 30 to 90 °C, with maximum activity at 65 °C, pH 5.0. The Km and Vmax determined using sodium polypectate were 6.9 mg·mL -1 and Vmax 690 μmol·min -1 .mg -1 , respectively. EplNg was active and stable over a wide range of pH values and temperatures, confirming the interesting properties EplNg and provide a basis for the development of the enzyme in different biotechnological processes., (Copyright © 2021 Elsevier B.V. All rights reserved.)

The wide distribution of the Neotropical freshwater catfish Rhamdia offers an excellent opportunity to investigate the historical processes responsible for modeling South America's hydrogeological structure. We used sequences from cis-Andean and Mesoamerican Rhamdia species to reconstruct and estimate divergence times among cis-Andean lineages, correlating the results with known geological events. Species delimitation methods based on distance (DNA barcoding and BIN) and coalescence (GMYC) approaches identified nine well-supported lineages from the cis-Andean region from sequences available in the BOLD dataset. The cis-Andean Rhamdia lineages diversification process began in Eocene and represented the split between cis-Andean and Mesoamerican clades. The cis-Andean clade contains two principal groups: Northwest clade (MOTUs from Amazon, Essequibo, Paraguay, and Itapecuru basins) and Southeast clade (Eastern Brazilian shield basins (Paraná, Uruguay, Iguaçu, and São Francisco) plus eastern coastal basins). The diversification of the cis-Andean Rhamdia lineages results from vicariance and geodispersion events, which played a key role in the current intricate distribution pattern of the Rhamdia lineages. The wide geographical distribution and large size of the specimens make it attractive to cultivate in different countries of the Neotropical region. The lineages delimitation minimizes identification mistakes, unintentional crossings by aquaculture, and reduces natural stocks contamination.

Aurora kinases are a family of serine/threonine protein kinases that play a central role in eukaryotic cell division. Overexpression of aurora kinases in cancer and their role as major regulators of the cell cycle quickly inspired the idea that their inhibition might be a potential pathway when treating oncologic patients. Over the past couple of decades, the search for designing and testing of molecules capable of inhibiting aurora activities fueled many pre-clinical and clinical studies. In this study, data from the past 10 years of in vitro and in vivo investigations, as well as clinical trials, utilizing aurora kinase inhibitors as therapeutics for hematological malignancies were compiled and discussed, aiming to highlight potential uses of these inhibitors as a novel monotherapy model or alongside conventional chemotherapies. While there is still much to be elucidated, it is clear that these kinases play a key role in oncogenesis, and their manageable toxicity and potentially synergistic effects still render them a focus of interest for future investigations in combinatorial clinical trials., Competing Interests: The Authors declare no conflicts of interest regarding this study., (Copyright 2021, International Institute of Anticancer Research.)

Background/aim: Experimental oncology commonly uses cells as oncological models, providing a framework for the testing of drugs, and investigation of cytotoxicity, mutagenesis and carcinogenesis. Investigations into poly-ADP-ribose polymerase 1 (PARP1) inhibition have become ever more relevant due to its approval as a therapeutic option for tumors with BRCA1/2 DNA repair-associated mutation and the seemingly high PARP expression levels in some tumor subtypes. In this study, we aimed to determine PARP1 gene expression of different hematological cancer-derived cell lineages and compare them to that of normal cell lines., Materials and Methods: PARP1 gene expression in seven different neoplastic lineages, representing three different hematological disorders (chronic myeloid leukemia, Burkitt lymphoma and acute lymphoblastic leukemia), was quantified by quantitative real-time polymerase chain reaction., Results: All hematological malignant lineages in this study overexpressed PARP1 when compared to the normal cell line MRC-5, with Burkitt's lymphoma cells having the highest expression values (fold change: 93)., Conclusion: Overexpression of PARP1 in hematological malignant lineages is a finding of crucial importance to future studies exploring possible cellular oncogenic pathways and supports investigations into the effectiveness of PARP1 inhibitors against hematological disorders., (Copyright © 2021 International Institute of Anticancer Research (Dr. George J. Delinasios), All rights reserved.)

Candida spp. is considered to be the third or fourth most common cause of bloodstream infections associated with healthcare services in the world. Currently, several strains exhibit resistance to the traditional treatments, making the development of new therapeutic molecules necessary. Drug repositioning is an alternative that can be used to work around problems such as toxicity, cost and time in the development of new drugs. This study aims to evaluate the in vitro antifungal effect of tropicamide, molecule of anticholinergic action, against planktonic cells of Candida spp. and biofilm of C. albicans. Six strains of different Candida species were used to determine the minimum inhibitory concentration (MIC) of tropicamide and fluconazole according to CLSI document M27-A3 and one strain of C. albicans was used to evaluate the activity of tropicamide against biofilms. In concentrations of 64μg/mL, the tropicamide exhibited 50% of inhibitory activity in planktonic cell and in concentrations of 128μg/mL is able to inhibit the formation of C. albicans biofilm. Despite the inhibitory activity shown at the present study, the use of a larger number of strains, as well as in vivo cytotoxicity assays, is necessary to confirm the hypothesis that tropicamide can be used as an adjuvant agent in the treatment of infections by the Candida genus., (Copyright © 2020 Elsevier Masson SAS. All rights reserved.)

The genus Schizodon is part of a group of headstanders and relatives (Family Anostomidae) that are widespread and ecologically important fishes in South American rivers. Schizodon includes 15 nominal species but their taxonomy has been challenging due to paucity of decisive characters to diagnose species. We present new molecular data to assess species boundaries or molecular operational taxonomic units (MOTUs), and to infer phylogenetic relationships among species. Evidence from two mitochondrial and three nuclear genes was used in these analyses. Mitochondrial DNA data for 112 specimens (from 11 nominal species) supported 13 consensus MOTUs, six of which matched valid nominal species (Schizodon borellii, S. fasciatus, S. intermedius, S. isognathus, S. knerii and S. scotorhabdotus). The nominal species Schizodon vittatus, S. nasutus, and S. dissimilis were subdivided into two MOTUs each, revealing either cryptic species or strong population structuring. In contrast, S. platae and S. jacuiensis constituted a single MOTU, indicating a possible case of synonymy. Our phylogenetic analysis subdivided the genus Schizodon into two large clades that are compatible with observed color patterns and biogeographic distribution. The first clade includes species with three to four conspicuous dark vertical bars on the flanks that originated in the Amazonas region (S. borellii, S. dissimilis, S. intermedius, S. fasciatus, S. scotorhabdotus, S. vittatus, and a cryptic species, Schizodon aff. vittatus). The second clade includes species with a conspicuous dark caudal blotch on the caudal peduncle, with vertical bars absent or inconspicuous, with a biogeographic origin in the La Plata drainage (S. isognathus, S. jacuiensis, S. knerii, S. nasutus and S. platae). Our results reinforce the importance of using molecular analyses to accelerate the study of diversity, particularly in groups with a wide distribution, few variable meristic characters, and high morphological plasticity, which may hide still unknown or underestimated diversity., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Telomeres compose the end portions of human chromosomes, and their main function is to protect the genome. In hematological disorders, telomeres are shortened, predisposing to genetic instability that may cause DNA damage and chromosomal rearrangements, inducing a poor clinical outcome. Studies from 2010 to 2019 were compiled and experimental studies using samples of patients diagnosed with hematological malignancies that reported the size of the telomeres were described. Abnormal telomere shortening is described in cancer, but in hematological neoplasms, telomeres are still shortened even after telomerase reactivation. In this study, we compared the sizes of telomeres in leukemias, myelodysplastic syndrome and lymphomas, identifying that the smallest telomeres are present in patients at relapse. In conclusion, the experimental and clinical data analyzed in this review demonstrate that excessive telomere shortening is present in major hematological malignancies and its analysis and measurement is a crucial step in determining patient prognosis, predicting disease risk and assisting in the decision for targeted therapeutic strategies., (Copyright© 2020, International Institute of Anticancer Research (Dr. George J. Delinasios), All rights reserved.)

The idea of utilizing poly-ADP-ribose polymerase inhibitors (PARPi) as therapeutics for cancer has grown in popularity since its original approval for clinical usage in treatment of BRCA DNA repair-associated-mutated ovarian cancer. In this study, we evaluated experimental data regarding in vitro studies utilizing PARPi as a treatment for tyrosine kinase (TK)-dependent leukemia. Studies from 2015 to 2019 were compiled and the ones with most relevant TK pathways and PARP inhibition were analyzed. PARPi showed activity against many leukemia cell lines and samples from patients with primary leukemia, especially when combined with other signaling pathway inhibitor drugs, improving upon the hypothesis that the utilization of PARPi has potential as a new therapeutic approach in treatment of primary leukemia and TK-dependent leukemia., (Copyright© 2020, International Institute of Anticancer Research (Dr. George J. Delinasios), All rights reserved.)

Motor neurons project axons from the hindbrain and spinal cord to muscle, where they induce myofibre contractions through neurotransmitter release at neuromuscular junctions. Studies of neuromuscular junction formation and homeostasis have been largely confined to in vivo models. In this study we have merged three powerful tools - pluripotent stem cells, optogenetics and microfabrication - and designed an open microdevice in which motor axons grow from a neural compartment containing embryonic stem cell-derived motor neurons and astrocytes through microchannels to form functional neuromuscular junctions with contractile myofibers in a separate compartment. Optogenetic entrainment of motor neurons in this reductionist neuromuscular circuit enhanced neuromuscular junction formation more than two-fold, mirroring the activity-dependence of synapse development in vivo. We incorporated an established motor neuron disease model into our system and found that coculture of motor neurons with SOD1G93A astrocytes resulted in denervation of the central compartment and diminished myofiber contractions, a phenotype which was rescued by the Receptor Interacting Serine/Threonine Kinase 1 (RIPK1) inhibitor Necrostatin. This coculture system replicates key aspects of nerve-muscle connectivity in vivo and represents a rapid and scalable alternative to animal models of neuromuscular function and disease., Competing Interests: Competing Interests The authors declare no competing interest.

Background: Metabolic syndrome (MetS) is a cluster of metabolic abnormalities that increases the cardiovascular risk. Regular physical exercise can promote benefits, but the MetS individuals are demotivated to perform it. Thus, new possibilities are important as an alternative intervention. The whole-body vibration can be considered an exercise modality and would be a safe and low-cost strategy to improve functional parameters of individuals in different clinical conditions. The aim of this exploratory study was to assess effects of whole-body vibration on functional parameters of MetS individuals. The hypothesis of this work was that the whole-body vibration could improve the functionality of MetS individuals., Methods: Twenty-two individuals performed the intervention. The vibration frequency varied from 5 to 14 Hz and the peak-to-peak displacements, from 2.5 to 7.5 mm. Each session consisted of one minute-bout of working time followed by a one minute-bout of passive rest in each peak-to-peak displacement for three-times. The whole-body vibration protocol was applied twice per week for 5 weeks. Data from the trunk flexion, gait speed, sit-to-stand test and handgrip strength were collected. Physiological parameters (blood pressure and heart rate) were also evaluated. The Wilcoxon Rank test and Student t-test were used., Results: No significant changes (p > 0.05) were observed in physiological parameters (arterial blood pressure and heart rate). Significant improvements were found in trunk flexion (p = 0.01), gait speed (p = 0.02), sit-to-stand test (p = 0.005) and handgrip strength (p = 0.04) after the whole-body vibration., Conclusions: In conclusion, whole-body vibration may induce biological responses that improve functional parameters in participants with MetS without interfering in physiological parameters, comparing before and after a 5-week whole-body vibration protocol., Trial Registration: Register in the Registro Brasileiro de Ensaios Clínicos (ReBEC) with the number RBR 2bghmh (June 6th, 2016) and UTN: U1111-1181-1177. (virgula).