Abstract Background: In recent decades, snake venom disintegrins have received special attention due to their potential use in anticancer therapy. Disintegrins are small and cysteine-rich proteins present in snake venoms and can interact with specific integrins to inhibit their activities in cell-cell and cell-ECM interactions. These molecules, known to inhibit platelet aggregation, are also capable of interacting with certain cancer-related integrins, and may interfere in important processes involved in carcinogenesis. Therefore, disintegrin from Crotalus durissus collilineatus venom was isolated, structurally characterized and evaluated for its toxicity and ability to interfere with cell proliferation and migration in MDA-MB-231, a human breast cancer cell line. Methods: Based on previous studies, disintegrin was isolated by FPLC, through two chromatographic steps, both on reversed phase C-18 columns. The isolated disintegrin was structurally characterized by Tris-Tricine-SDS-PAGE, mass spectrometry and N-terminal sequencing. For the functional assays, MTT and wound-healing assays were performed in order to investigate cytotoxicity and effect on cell migration in vitro, respectively. Results: Disintegrin presented a molecular mass of 7287.4 Da and its amino acid sequence shared similarity with the disintegrin domain of P-II metalloproteases. Using functional assays, the disintegrin showed low cytotoxicity (15% and 17%, at 3 and 6 μg/mL, respectively) after 24 h of incubation and in the wound-healing assay, the disintegrin (3 μg/mL) was able to significantly inhibit cell migration (24%, p < 0.05), compared to negative control. Conclusion: Thus, our results demonstrate that non-RGD disintegrin from C. d. collilineatus induces low cytotoxicity and inhibits migration of human breast cancer cells. Therefore, it may be a very useful molecular tool for understanding ECM-cell interaction cancer-related mechanisms involved in an important integrin family that highlights molecular aspects of tumorigenesis. Also, non-RGD disintegrin has potential to serve as an agent in anticancer therapy or adjuvant component combined with other anticancer drugs.

RESUMEN Objetivo Evaluar el Programa “Mais Médicos” en el Brasil mediante el cálculo del aumento proporcional del número de médicos en los municipios participantes y los costos del programa, con estratificación por componente del costo y fuente de financiamiento. Métodos Se utilizaron datos oficiales de la edición del 2013 de Demografia médica no Brasil [demografía médica en el Brasil] para calcular el número de médicos antes del Programa “Mais Medicos”. El número de médicos al final del cuarto ciclo de reclutamiento del programa (julio del 2014) se obtuvo del Ministerio de Salud. Se determinaron y calcularon los componentes de costos sobre la base de la legislación y las directrices del programa. Se eligieron los municipios participantes según cuatro criterios, todos relacionados con la vulnerabilidad. Resultados Mediante el Programa “Mais Médicos” se destinaron 14 462 médicos más a zonas sumamente vulnerables y remotas en 3 785 municipios (68% del total) y 34 distritos sanitarios indígenas especiales. El aumento del número de médicos fue mayor en las zonas más pobres (al Norte y Nordeste). El costo anual estimado de US$ 1 100 millones incluyó la dotación de médicos, la educación continua y la supervisión y tutoría. El financiamiento estuvo principalmente centralizado en el nivel federal (92,6%). Conclusiones Se considera que el costo del Programa “Mais Médicos” es relativamente moderado en relación con los posibles beneficios para la salud de la población. Con el mayor aumento de la dotación de médicos en las poblaciones más pobres y vulnerables se cumplió el objetivo de corregir los desequilibrios en la distribución del personal de salud. La experiencia de este programa en el Brasil puede contribuir al debate sobre cómo paliar la escasez de médicos.

RESUMO Objetivo Avaliar o Programa Mais Médicos (PMM) no Brasil estimando o aumento proporcional do número de médicos nos municípios participantes e os custos do programa, estratificado pelo componente de custo e fonte de financiamento. Métodos Os dados oficiais da edição de Demografia médica no Brasil de 2013 foram usados para estimar o número de médicos anterior ao PMM. O número de médicos ao final do quarto ciclo de recrutamento do PMM (julho de 2014) foi fornecido pelo Ministério da Saúde. Os componentes de custo foram identificados e calculados de acordo com a legislação e as diretrizes do PMM. Os municípios participantes foram selecionados segundo quatro critérios relacionados à vulnerabilidade. Resultados O PMM proveu um adicional de 14.462 médicos para áreas remotas bastante vulneráveis em 3.785 municípios (68% do total) e 34 distritos sanitários especiais indígenas. Houve aumento maior do número de médicos nas regiões mais pobres (Norte e Nordeste). O custo anual estimado de US$ 1,1 bilhão cobriu provisões médicas, educação continuada e supervisão/mentoria. O financiamento foi em grande parte centralizado ao nível federal (92,6%). Conclusão Considera-se que o custo do PMM seja relativamente moderado em relação aos benefícios em potencial à saúde da população. O aumento maior no número de médicos para atender a população mais pobres e vulnerável alcançou a meta de corrigir a má distribuição de profissionais da saúde. A experiência do PMM no Brasil pode subsidiar o debate visando reduzir a escassez de médicos.

ABSTRACT Objective To evaluate the Programa Mais Médicos (More Doctors Program; PMM) in Brazil by estimating the proportional increase in the number of doctors in participating municipalities and the program costs, stratified by cost component and funding source. Methods Official data from the 2013 edition of Demografia Médica no Brasil (Medical Demography in Brazil) was used to estimate the number of doctors prior to PMM. The number of doctors at the end of the fourth PMM recruiting cycle (July 2014) was obtained from the Ministry of Health. Cost components were identified and estimated based on PMM legislation and guidelines. The participating municipalities were chosen based on four criteria, all related to vulnerability. Results The PMM provided an additional 14 462 physicians to highly vulnerable, remote areas in 3 785 municipalities (68% of the total) and 34 Special Indigenous Sanitary Districts. There was a greater increase of physicians in the poorest regions (North and Northeast). The estimated annual cost of US$ 1.1 billion covered medical provision, continuing education, and supervision/mentoring. Funding was largely centralized at the federal level (92.6%). Conclusion The cost of PMM is considered relatively moderate in comparison to its potential benefits for population health. The greater increase of doctors for the poorest and most vulnerable met the target of correcting imbalances in health worker distribution. The PMM experience in Brazil can contribute to the debate on reducing physician shortages.

Abstract Background In Brazil, accidents with venomous animals are considered a public health problem. Tityus serrulatus (Ts), popularly known as the yellow scorpion, is most frequently responsible for the severe accidents in the country. Ts envenoming can cause several signs and symptoms classified according to their clinical manifestations as mild, moderate or severe. Furthermore, the victims usually present biochemical alterations, including hyperglycemia. Nevertheless, Ts envenoming and its induced hyperglycemia were never studied or documented in a patient with diabetes mellitus (DM). Therefore, this is the first study to evaluate the glycemia during Ts envenoming using a diabetic animal model (NOD, non-obese diabetic). Methods Female mice (BALB/c or NOD) were challenged with a non-lethal dose of Ts venom. Blood glucose level was measured (tail blood using a glucose meter) over a 24-h period. The total glycosylated hemoglobin (HbA1c) levels were measured 30 days after Ts venom injection. Moreover, the insulin levels were analyzed at the glycemia peak. Results The results demonstrated that the envenomed NOD animals presented a significant increase of glycemia, glycosylated hemoglobin (HbA1c) and insulin levels compared to the envenomed BALB/c control group, corroborating that DM victims present great risk of developing severe envenoming. Moreover, the envenomed NOD animals presented highest risk of death and sequelae. Conclusions This study demonstrated that the diabetic victims stung by Ts scorpion should be always considered a risk group for scorpion envenoming severity.

Abstract Background In the Atlantic forest of the North and Northeast regions of Brazil, local population often uses the fruit juice and the aqueous extract of leaves of soursop (Annona muricata L.) to treat Lachesis muta rhombeata envenomation. Envenomation is a relevant health issue in these areas, especially due to its severity and because the production and distribution of antivenom is limited in these regions. The aim of the present study was to evaluate the relevance of the use of soursop leaf extract and its juice against envenomation by Lachesis muta rhombeata. Methods We evaluated the biochemical, hematological and hemostatic parameters, the blood pressure, the inflammation process and the lethality induced by Lachesis muta rhombeata snake venom. We also assessed the action of the aqueous extract of leaves (AmL) and juice (AmJ) from A. muricata on the animal organism injected with L. m. rhombeata venom (LmrV) in the laboratory environment. Results LmrV induced a decrease of total protein, albumin and glucose; and increase of creatine kinase, aspartate aminotransferase, and urea concentrations. It provoked hemoconcentration followed by reduction of hematocrit, an increase in prothrombin time and partial thromboplastin time and a decrease of the blood pressure. LmrV induced the release of interleukin-6, an increase in neutrophils and changes in the serum protein profile, characteristic of the acute inflammatory process. LD50 values were similar for the groups injected with LmrV and treated or untreated with AmJ and AmL. Both treatments play a role on the maintenance of blood glucose, urea and coagulation parameters and exert a protective action against the myotoxicity. However, they seem to worsen the hypotension caused by LmrV. Conclusion The treatments with AmJ and AmL present some beneficial actions, but they might intensify some effects of the venom. Therefore, additional studies on A. muricata are necessary to enable its use as natural antivenom for bushmaster snakebite.

The phytopathogenic fungus Moniliophthora perniciosa (Stahel) Aime & Philips-Mora, causal agent of witches' broom disease of cocoa, causes countless damage to cocoa production in Brazil. Molecular studies have attempted to identify genes that play important roles in fungal survival and virulence. In this study, sequences deposited in the M. perniciosa Genome Sequencing Project database were analyzed to identify potential biological targets. For the first time, the ergosterol biosynthetic pathway in M. perniciosa was studied and the lanosterol 14α-demethylase gene (ERG11) that encodes the main enzyme of this pathway and is a target for fungicides was cloned, characterized molecularly and its phylogeny analyzed. ERG11 genomic DNA and cDNA were characterized and sequence analysis of the ERG11 protein identified highly conserved domains typical of this enzyme, such as SRS1, SRS4, EXXR and the heme-binding region (HBR). Comparison of the protein sequences and phylogenetic analysis revealed that the M. perniciosa enzyme was most closely related to that of Coprinopsis cinerea.