Identification of fish specimens of the Tocantins River, Brazil, using DNA barcoding.

The fish fauna of the Tocantins River possesses many endemic species; however, it is little studied in molecular terms and is quite threatened by the construction of several hydroelectric dams. Therefore, the objective of this study was to identify the ichthyofauna of the Tocantins River using DNA barcoding. For this, collections were carried out in five points of this river, which resulted in the capture of 725 individuals from which partial sequences of the cytochrome oxidase subunit I (COI) gene were obtained for genetic analysis. A total of 443 haplotypes were recovered with the mean intraspecific K2P genetic distance of 1.82%. Altogether, 138 species were identified based on morphological criteria, which was a quantity that was much lower than that indicated by the four molecular methods (assemble species by automatic partitioning [ASAP], barcode index number [BIN], generalized mixed Yule coalescent (GMYC), and Bayesian Poisson tree processes [bPTP]) through which 152-157 molecular entities were identified. In all, 41 unique BINs were obtained based on the data generated in the BOLDSystems platform. According to the result indicated by ASAP (species delimitation approach considered the most appropriate in the present study), there was an increase of 17 molecular entities (12.32%), when compared to the number of species identified through their morphological criteria, as it can show cryptic diversity, candidates for new species, and misidentifications. There were 21 incongruities indicated between the different identification approaches for species. Therefore, it is suggested that these taxonomic problems be cautiously evaluated by experts to solve such taxonomic issues.

The Health Status of Horses Used for at Least Six Complete Cycles of Loxoscelic Antivenom Production.

Antivenom production against Loxosceles venom relies on horses being immunized and bled for plasma harvest. One horse can partake in several cycles of antivenom production, which will require years of constant venom and adjuvant inoculation and bleeding. The actual impact on the health of horses that participate in several antivenom-producing cycles is unknown. Therefore, this study aimed to evaluate the general health status of horses that underwent at least six cycles of loxoscelic antivenom production. Seven crossbred horses that had partaken in six to eight complete antivenom-producing cycles were used and established as the immunized group (IG). Under the same handling and general management, eleven horses were established as the control group (CG). The horses were evaluated regarding their general clinical status and had their blood sampled, and an ECG recorded. The IG presented lower RBC and PCV, despite keeping values within inferior limits for the species. Renal function was not impaired, and liver-related enzymes were higher than those in the CG, probably due to liver exertion from immunoglobulin synthesis. ECG showed some abnormalities in the IG, such as atrioventricular block and a wandering atrial pacemaker, corroborated by an increase in CK-MB. The cardiovascular abnormalities were mainly found in the horses that participated in several antivenom-producing cycles. The overall results indicate that these horses had some impairment of their general health status. Once available, some alternative, less toxic antigens should replace the venom for immunization of horses used for antivenom production.

Prevalence of childhood anemia: Potential sociodemographic and dietary factors in Nigeria.

BACKGROUND: Childhood anemia is a global public health issue. In this study, we assessed the potential sociodemographic and dietary factors associated with the prevalence of anemia among children aged 6-59 months in Nigeria. METHODS: In this cross-sectional study, we collected dietary information and demographic data on 6,338 children with anemia from the Nigerian Demographic and Health Surveys (2018). The association between the occurrence of anemia and the demographic and dietary factors was determined by conducting Chi-squared tests. Additionally, bivariate and multivariate order logit models were constructed and reported as odds ratios. RESULTS: The results of the multivariate analysis showed that the risk of anemia was reduced by 13% and 44% in children aged 13-36 months (OR = 0.87; 95% CI = 0.77-0.98; p = 0.019) and 37-59 months (OR = 0.56; 95% CI = 0.49-0.63; p < 0.001), respectively, compared to the risk of anemia in children aged 6-12 months. Anemia was 28% less likely in children of non-anemic mothers (OR = 0.72; 95% CI = 0.66-0.80; p < 0.001) than children of anemic mothers. Children fed pumpkin, carrot, squash, and sweet potato showed a lower occurrence of anemia by 17% (OR = 0.83; 95% CI = 0.70-0.99; p = 0.036) compared to those who were not fed these vegetables. Chances of anemia increased by 14% in children who were fed white potatoes, white yams, manioc, cassava, and other root-based foods (OR = 1.14; 95% CI = 1.01-1.29; p = 0.036). CONCLUSION: This study highlighted the impact of a plant-based diet on the high prevalence of childhood anemia in Nigeria. Therefore, reformation of dietary habits, the inclusion of nutritional supplements, and food-fortification programs with reductions in maternal anemia are recommended.

Lung Inflammasome Activation in SARS-CoV-2 Post-Mortem Biopsies.

The inflammasome complex is a key part of chronic diseases and acute infections, being responsible for cytokine release and cell death mechanism regulation. The SARS-CoV-2 infection is characterized by a dysregulated cytokine release. In this context, the inflammasome complex analysis within SARS-CoV-2 infection may prove beneficial to understand the disease's mechanisms. Post-mortem minimally invasive autopsies were performed in patients who died from COVID-19 (n = 24), and lung samples were compared to a patient control group (n = 11) and an Influenza A virus H1N1 subtype group from the 2009 pandemics (n = 10). Histological analysis was performed using hematoxylin-eosin staining. Immunohistochemical (IHC) staining was performed using monoclonal antibodies against targets: ACE2, TLR4, NF-κB, NLRP-3 (or NALP), IL-1ß, IL-18, ASC, CASP1, CASP9, GSDMD, NOX4, TNF-α. Data obtained from digital analysis underwent appropriate statistical tests. IHC analysis showed biomarkers that indicate inflammasome activation (ACE2; NF-κB; NOX4; ASC) were significantly increased in the COVID-19 group (p < 0.05 for all) and biomarkers that indicate cell pyroptosis and inflammasome derived cytokines such as IL-18 (p < 0.005) and CASP1 were greatly increased (p < 0.0001) even when compared to the H1N1 group. We propose that the SARS-CoV-2 pathogenesis is connected to the inflammasome complex activation. Further studies are still warranted to elucidate the pathophysiology of the disease.

Booklet for knowledge and prevention of HIV mother-to-child transmission: a pilot study of a randomized clinical trial.

OBJECTIVE: To test the effectiveness of the booklet, compared to the usual service care, in the increase of the knowledge of pregnant/puerperal women living with HIV, for the prevention of HIV-VT. METHOD: Pilot study of a randomized controlled clinical trial, initially with 104 pregnant women living with HIV, with a final sample of 45 women. It was held in three public maternity hospitals in Fortaleza-CE, from January/2017 to May/2018. The control group received regular care from the service and the intervention group had access to the booklet as an additive. The research was carried out in three phases: baseline; evaluation 2, in prenatal care; and evaluation 3, in the postpartum period. RESULTS: There was no intergroup difference in the women's mean knowledge score (short-term p = 0.473; long-term p = 0.151). However, in the intragroup analysis, the booklet proved to be effective in improving the pregnant women's knowledge in the intervention group, in the short term (p = 0.002) and long term (p = 0.033). CONCLUSION: There was an improvement in knowledge within the intervention group over time, but there was no difference in women's knowledge in the intergroup analysis. Thus, based on this pilot, a broader study on the use of booklet is required to prove its effectiveness (ReBEC: UTN: U1111-1191-9954).

Analysis of NGS data from Peruvian Loxosceles laeta spider venom gland reveals toxin diversity.

Accidents involving spiders from the genus Loxosceles cause medical emergencies in several countries of South America. The species Loxosceles laeta is ubiquitously present in Peru and is responsible for severe accidents in this country. To further characterize L. laeta venom components and to unveil possible variations in the Peruvian population, we provide an overview of the toxins-related transcripts present in the venom gland of Peruvian L. laeta. A dataset from a cDNA library previously sequenced by MiSeq sequencer (Illumina) was re-analyzed and the obtained data was compared with available sequences from Loxosceles toxins. Phospholipase-D represent the majority (69,28 %) of the transcripts related to venom toxins, followed by metalloproteases (20,72 %), sicaritoxins (6,03 %), serine-proteases (2,28 %), hyaluronidases (1,80 %) and Translationally Controlled Tumor Protein (TCTP) (0,56 %). New sequences of phospholipases D,sicaritoxins, hyaluronidase, TCTP and serine proteinases were described. Differences between the here-described toxin sequences and others, previously identified in venom glands from other spiders, were visualized upon sequence alignments. In addition, an in vitro hyaluronidase activity assay was also performed to complement comparisons between Peruvian and Brazilian L. laeta venom enzymatic activities, revealing a superior activity in the venom from Brazilian specimens. These new data provide a molecular basis that can help to explain the difference in toxicity among L. laeta venoms from different countries in South America.

Protective Effectiveness of an Immunization Protocol Against the Toxic Effects of Loxosceles intermedia Venom in Rabbits.

Loxosceles spp. (brown spiders) bites are responsible for the development of a syndrome consisting mainly of dermonecrotic lesions, and also systemic effects. Rabbits are one of the main experimental models used for better understanding the systemic and local effects of Loxosceles venom. The aim of this study is to evaluate the toxic and protective effects of rabbits immunized with Loxosceles spp. venom. Male New Zealand rabbits were allocated as a control group (CG; n = 5) that received adjuvant (Montanide) and phosphate-buffer saline (PBS), or as venom group (VG; n = 5) that received 21 µg of Loxosceles venom using Montanide as adjuvant. After five immunization cycles, a trial with 7 µg of Loxosceles intermedia (L. intermedia) venom was performed, and dermonecrotic lesions were measured. The rabbits were then euthanized, and their organs were collected for histopathology analysis. Rabbits that had undergone Loxosceles venom immunization protocol showed minor clinical disturbances during the experimental period. The used immunization protocol protected the rabbits against the toxic effect of the Loxosceles venom because they showed minor clinical disturbances during the experimental period.

Clinical Effects of the Immunization Protocol Using Loxosceles Venom in Naïve Horses.

Bites of brown spiders (Loxosceles spp.) are responsible for dermonecrotic lesions and potentially systemic envenoming that can lead to death. The only effective therapy is the use of the antivenom, usually produced in horses. However, little is known about the consequences of the systematic use of the Loxosceles venom and adjuvants and of the bleedings on antivenom-producing horses. Thus, the aim of this study was to evaluate the clinical changes in horses in their first immunization protocol for Loxosceles antivenom production. Eleven healthy horses, never immunized, were evaluated in three different periods: T0 (before immunization); T1 (after their first venom immunization); and T2 (after their first bleeding). Horses were clinically evaluated, sampled for blood, and underwent electrocardiographic (ECG) recordings. Several suppurated subcutaneous abscesses occurred due to the use of Freund's adjuvants and thrombophlebitis due to systematic venipunctures for the bleeding procedures. ECG showed arrhythmias in few horses in T2, such as an increase in T and R waves. In summary, the immunization protocol impacted on horses' health, especially after bleeding for antivenom procurement.

Perfil demográfico y epidemiológico de los casos de dengue en un municipio de Brasil / Demographic and Epidemiological Profile of Dengue Cases in a Municipality from Brazil

Introducción: Las epidemias de dengue imponen una carga importante a los servicios de salud, además de la pérdida económica ocasionada por bajas temporales del trabajo o por años de vidas perdidas como consecuencia de la muerte, lo que interfiere en el desempeño de las economías de los países. Objetivo: Analizar el perfil demográfico, social y epidemiológico de los casos de dengue en la población de Contagem-MG, además de evaluar los factores climáticos con el fin de brindar herramientas para la planificación de estrategias de salud pública. Método: Se realizó un estudio ecológico, a partir de una serie de tiempo, entre los años 2013 a 2017, en el municipio de Contagem-MG en Brasil. Resultados: Los grupos más afectados fueron las mujeres, adultos, mestizos, individuos con pocos años de estudio y los que vivían en regiones periféricas. Más del 99 por ciento de las personas tenía dengue sin signos de alarma. Las lluvias no parecen haber sido determinantes para el aumento del número de casos, dado que hubo lluvias en todos los años. Conclusiones: Para mejorar la lucha contra las epidemias estacionales de dengue, se sugiere el enfoque de planificación estratégica, con la elección y definición de metodologías para planificar una estrategia intersectorial en la solución de situaciones recurrentes. Así se puede incrementar la eficiencia de las políticas públicas y, por ende, las posibilidades de una gestión pública exitosa en la reducción de inequidades(AU)

Introduction: Dengue epidemics impose a significant burden on health services, in addition to the economic loss caused by temporary leaves from work or deaths, which interferes in the economic performance of countries. Objective: To analyze the demographic, social and epidemiological profile of dengue cases in the population of Contagem-MG; and to evaluate climatic factors in order to provide tools for planning public health strategies. Methods: An ecological study was conducted from a time series between 2013 and 2017 in Contagem-MG municipality from Brazil. Results: The most affected groups were women, adults, mestizos, people with few years of schooling, and those living in peripheral regions. More than 99 percent of the people suffered from dengue without warning signs. Rain does not seem to be a determinant factor in the increase on the number of cases, since it rained every year. Conclusions: To better fight against dengue seasonal epidemics, the strategic planning approach is suggested, with the selection and definition of methodologies for planning an intersectoral strategy to solve recurrent events. This could increase the efficiency of public policies and, therefore, the possibility of successful public management in the reduction of inequities(AU)

Leishmania diversity in bats from an endemic area for visceral and cutaneous leishmaniasis in Southeastern Brazil.

This study aimed to determine the occurrence of Leishmania infection in bats in urban and wild areas in an endemic municipality for visceral and cutaneous leishmaniasis in Minas Gerais, Brazil. Between April 2014 to April 2015, 247 bats were captured and classified into 26 species belonging to Phyllostomidae (90.7%), Vespertilionidae (8.1%) and Molossidae (1.2%) families. Blood samples from 247 bats were collected and submitted to nested-PCR, targeting the variable V7-V8 region of the SSU rRNA gene, followed by sequencing of the PCR product. The overall infection rate of Leishmania spp. in bats was 4.4%. Of the eleven bats infected, ten were frugivorous bats: Artibeus planirostris (8/11), Artibeus lituratus (1/11) and Artibeus cinereus (1/11) and one a nectarivorous bat (Glossophaga soricina). None of the individuals exhibited macroscopic alterations in the skin, spleen or liver. Phylogenetic analysis separated Leishmania species in clades corresponding to the subgenera Viannia, Leishmania, and Mundinia, and supported that the isolates characterized in the present study clustered closely with Leishmania (Viannia) sp., Leishmania (Leishmania) infantum and Leishmania (Leishmania) amazonensis. Here we report for the first time the bat Artibeus cinereus as a host of Leishmania (Leishmania) amazonensis. In the study we found that the mean abundance of bats did not differ in wild habitats and urban areas and that bat-parasite interactions were similarly distributed in the two environments. On the other hand, further studies should be conducted in more recent times to verify whether there have been changes in these parameters.

Booklet for knowledge and prevention of HIV mother-to-child transmission: a pilot study of a randomized clinical trial / Cuadernillo para el conocimiento y la prevención de la transmisión maternoinfantil del VIH: un estudio piloto de un ensayo clínico aleatorizado / Cartilha para conhecimento e prevenção da transmissão vertical do HIV: estudo piloto de ensaio clínico randomizado

ABSTRACT Objective: To test the effectiveness of the booklet, compared to the usual service care, in the increase of the knowledge of pregnant/puerperal women living with HIV, for the prevention of HIV-VT. Method: Pilot study of a randomized controlled clinical trial, initially with 104 pregnant women living with HIV, with a final sample of 45 women. It was held in three public maternity hospitals in Fortaleza-CE, from January/2017 to May/2018. The control group received regular care from the service and the intervention group had access to the booklet as an additive. The research was carried out in three phases: baseline; evaluation 2, in prenatal care; and evaluation 3, in the postpartum period. Results: There was no intergroup difference in the women's mean knowledge score (short-term p = 0.473; long-term p = 0.151). However, in the intragroup analysis, the booklet proved to be effective in improving the pregnant women's knowledge in the intervention group, in the short term (p = 0.002) and long term (p = 0.033). Conclusion: There was an improvement in knowledge within the intervention group over time, but there was no difference in women's knowledge in the intergroup analysis. Thus, based on this pilot, a broader study on the use of booklet is required to prove its effectiveness (ReBEC: UTN: U1111-1191-9954).

RESUMEN Objetivo: Testar la eficacia del cuardenillo, en comparación al servicio habitual, en el incremento del conocimiento de las mujeres embarazadas/puérperas que viven con el VIH, para la prevención del TV-VIH. Método: Piloto de un ensayo clínico controlado aleatorizado, inicialmente con 104 mujeres embarazadas que viven con el VIH, con una muestra final de 45 mujeres. Realizado en tres maternidades públicas de Fortaleza-CE, de enero/2017 a mayo/2018. El grupo control recibió atención regular del servicio y el grupo intervención tuvo acceso al cuadernillo como complemento. La investigación se llevó a cabo en tres etapas: línea de base; evaluación 2, en atención prenatal; y evaluación 3, en el puerperio. Resultados: No hubo diferencia entre grupos en la puntuación media de conocimiento de las mujeres (a corto plazo p = 0,473; a largo plazo p = 0,151). Sin embargo, en el análisis intragrupo, el cuardenillo se mostró eficaz en la mejora del conocimiento de las gestantes del grupo intervención, a corto plazo (p = 0,002) y a largo plazo (p = 0,033). Conclusión: Hubo una mejora en el conocimiento dentro del grupo de intervención con el tiempo, pero no hubo diferencia en el conocimiento de las mujeres en el análisis intergrupal. Por lo tanto, el cuadernillo necesita realizar un estudio más amplio, basado en este piloto, para probar su efectividad (ReBEC: UTN: U1111-1191-9954).

RESUMO Objetivo: Testar efetividade de cartilha, em comparação ao atendimento habitual do serviço, no aumento do conhecimento de gestantes/puérperas que vivem com HIV, para prevenção da TV-HIV. Método: Piloto de ensaio clínico randomizado controlado, inicialmente com 104 gestantes que vivem com HIV, com amostra final de 45 mulheres. Realizado em três maternidades públicas de Fortaleza-CE, de janeiro/2017 a maio/2018. O grupo controle recebeu atendimento habitual do serviço e o grupo intervenção teve como aditivo acesso à cartilha. A pesquisa foi realizada em três fases: linha de base; avaliação 2, no pré-natal; e avaliação 3, no pós-parto. Resultados: Não houve diferença intergrupos na média da pontuação do conhecimento das mulheres (curto prazo p = 0,473; longo prazo p = 0,151). Porém, na análise intragrupo, a cartilha se mostrou efetiva para melhorar o conhecimento das gestantes do grupo intervenção, em curto prazo (p = 0,002) e longo prazo (p = 0,033). Conclusão Houve melhora do conhecimento dentro do grupo intervenção, ao longo do tempo, porém não foi evidenciada diferença quanto ao conhecimento das mulheres na análise intergrupos. Assim, a cartilha carece da realização de estudo mais amplo, a partir deste piloto, para comprovação de sua efetividade (ReBEC: UTN: U1111-1191-9954).

A prokaryote system optimization for rMEPLox expression: A promising non-toxic antigen for Loxosceles antivenom production.

Loxoscelism is the most dangerous araneism form in Brazil and antivenom therapy is the recommended treatment. Antivenom is produced by horse immunization with Loxosceles spider venom, which is toxic for the producer animal. Moreover, due to the high amount of venom required for horse hyperimmunization, new strategies for antigens obtention have been proposed. In this sense, our research group has previously produced a non-toxic recombinant multiepitopic protein derived from Loxosceles toxins (rMEPLox). rMEPLox was a successful immunogen, being able to induce the production of neutralizing antibodies, which could be used in the Loxoscelism treatment. However, rMEPLox obtention procedure requires optimization, as its production needs to be scaled up to suit antivenom manufacture. Therefore, an effective protocol development for rMEPlox production would be advantageous. To achieve this objective, we evaluated the influence of different cultivation conditions for rMEPLox optimum expression. The optimum conditions to obtain large amounts of rMEPlox were defined as the use of C43(DE3)pLysS as a host strain, 2xTY medium, 0.6 mM IPTG, biomass pre induction of OD600nm = 0.4 and incubation at 30 °C for 16 h. Following the optimized protocol, 39.84 mg/L of soluble rMEPLox was obtained and tested as immunogen. The results show that the obtained rMEPLox preserved the previously described immunogenicity, and it was able to generate antibodies that recognize different epitopes of the main Loxosceles venom toxins, which makes it a promising candidate for the antivenom production for loxoscelism treatment.

History, challenges and perspectives on Loxosceles (brown spiders) antivenom production in Brazil.

Antivenom is the only effective therapy for treating any envenomation. Despite its obvious public health importance, the laborious process of procuring, distributing and controlling the quality of such immunobiologicals is being neglected. Brazil is fully self-sufficient in the production of antivenoms. Since the 1950s, Loxoscelism, a syndrome with an onset after a spider bite from specimens of the Loxosceles genus occurs, is considered a public health issue. The Brazilian history in developing antivenom therapy, its production hindrances, and other challenges are discussed in this paper, as well as some promising novelties that can improve production and processing of Loxosceles antivenom.

Genetic and toxinological divergence among populations of Tityus trivittatus Kraepelin, 1898 (Scorpiones: Buthidae) inhabiting Paraguay and Argentina.

Envenoming by scorpions in genus Tityus is a public health problem in Tropical America. One of the most medically significant species is Tityus trivittatus, which is known to occur from southwest Brazil to central-northern and eastern Argentina. In this work, we studied the lethality, composition, antigenicity, and enzymatic activity of venom from a T. trivittatus population found further north in urban areas of eastern Paraguay, where it has caused serious envenomation of children. Our results indicate that the population is of medical importance as it produces a potently toxic venom with an LD50 around 1.19 mg/kg. Venom neutralization in preliminary mouse bioassays was complete when using Brazilian anti-T. serrulatus antivenom but only partial when using Argentinean anti-T. trivittatus antivenom. Venom competitive solid-phase enzyme immunoassays and immunoblotting from Argentinean and Paraguayan T. trivittatus populations indicated that antigenic differences exist across the species range. SDS-PAGE showed variations in type and relative amounts of venom proteins between T. trivitattus samples from Argentina and Paraguay. MALDI-TOF mass spectrometry indicated that while some sodium channel toxins are shared, including ß-toxin Tt1g, others are population-specific. Proteolytic activity by zymography and peptide identification through nESI-MS/MS also point out that population-specific proteases may exist in T. trivitattus, which are postulated to be involved in the envenoming process. A time-calibrated molecular phylogeny of mitochondrial COI sequences revealed a significant (8.14%) genetic differentiation between the Argentinean and Paraguayan populations, which appeared to have diverged between the mid Miocene and early Pliocene. Altogether, toxinological and genetic evidence indicate that T. trivitattus populations from Paraguay and Argentina correspond to distinct, unique cryptic species, and suggest that further venom and taxonomic diversity exists in synanthropic southern South American Tityus than previously thought.

Engineered antigen containing epitopes from Loxosceles spp. spider toxins induces a monoclonal antibody (Lox-mAb3) against astacin-like metalloproteases.

Loxoscelism pose a health issue in the South America. The treatment for these accidents is based on the administration of antivenom produced in animals immunized with Loxosceles venom. In this work, a previously produced non-toxic multiepitopic chimeric protein (rMEPlox), composed of epitopes derived from the main toxins families (sphyngomielinase-D, metalloproteases, and hyaluronidases) of Loxosceles spider venoms, was used as antigen to produce monoclonal antibodies (mAbs). A selected anti-rMEPlox mAb (Lox-mAb3) reacted with metalloprotease from L. intermedia venom and showed cross-reactivity with metalloproteses from Brazilian and Peruvian Loxosceles laeta and Loxosceles gaucho venoms in immunoassays. The sequence recognized by Lox-mAb3 (184ENNTRTIGPFDYDSIMLYGAY205) corresponds to the C-terminal region of Astacin-like metalloprotease 1 and the amino acid sequence IGPFDYDSI, conserved among the homologs metalloproteases sequences, is important for antibody recognition. Lox-mAb3 neutralizes the fibrinogenolytic activity caused by metalloprotease from L. intermedia spider venom in vitro, which may lead to a decrease in hemorrhagic disturbances caused by Loxosceles envenomation. Our results show, for the first time, the use of a non-toxic multiepitopic protein for the production of a neutralizing monoclonal antibody against a metalloprotease of medically important Loxosceles venoms. These results contribute for the production improvement of therapeutic antivenom against loxoscelism.

Immunoprotection against lethal effects of Crotalus durissus snake venom elicited by synthetic epitopes trapped in liposomes.

Snakebites caused by Crotalus genus are the second most frequent in Brazil. Crotoxin is a beta-neurotoxin responsible for the main envenomation effects of Crotalus biting, while crotamine immobilizes the animal hind limbs, contributing to prey immobilization and to envenoming symptoms. As crotoxin and crotamine represent about 90% of Crotalus venom dry weight, these toxins are of great importance for antivenom therapy. In this sense, knowledge regarding the antigenicity/immunogenicity at the molecular level of these toxins can provide valuable information for the improvement of specific antivenoms. Therefore, the aims of this study are the identification of the B-cell epitopes from crotoxin and crotamine; and the characterization of the neutralizing potency of antibodies directed against the corresponding synthetic epitopes defined in the current study. Linear B-cell epitopes were identified using the Spot Synthesis technique probed with specific anti-C. d. terrificus venom horse IgG. One epitope of crotamine (F12PKEKICLPPSSDFGKMDCRW32) and three of crotoxin (L10LVGVEGHLLQFNKMIKFETR30; Y43CGWGGRGRPKDATDRCCFVH63 and T118YKYGYMFYPDSRCRGPSETC138) were identified. After synthesis in their soluble form, the peptides mixture correspondent to the mapped epitopes was entrapped in liposomes and used as immunogens for antibody production in rabbits. Anti-synthetic peptide antibodies were able to protect mice from the lethal activity of C. d. terrificus venom.

Comparação das respostas fisiológicas do incremental shuttle walk test e do Glittre-ADL test: instrumentos complementares para avaliação da capacidade funcional / Comparación de las respuestas fisiológicas de incremental shuttle walk test y de Glittre-ADL test: instrumentos complementarios para evaluar la capacidad funcional / Comparison of the physiological responses of the Incremental Shuttle Walk test and Glittre-ADL test: complementary instruments for the evaluation of functional capacity

RESUMO O objetivo deste estudo foi avaliar e comparar respostas metabólicas, cardiovasculares e ventilatórias do incremental shuttle walk test (ISWT) e do Glittre activities of daily living test (Glittre-ADL test). Trata-se de um estudo transversal, realizado com indivíduos saudáveis. A capacidade funcional (CF) foi avaliada pela distância percorrida e consumo de oxigênio pico (VO2pico) no ISWT e pelo tempo gasto e VO2 no Glittre-ADL test. Trinta indivíduos percorreram 656,67 (IC95%:608,8-704,5) metros no ISWT e executaram o Glittre-ADL test em 2,4 (IC95%:2,2-2,6) minutos. O VO2 pico do ISWT foi 27,8 (IC95%25,6-29,9) versus 22,2 (IC95%20,5-24,1)mL×kg−1×min−1 (p<0,001) no estado estável (EE) do Glittre-ADL test. As correlações entre distância percorrida no ISWT e o tempo gasto no Glittre-ADL test, o VO2pico do ISWT e o VO2 no EE do Glittre-ADL test e a FC no pico do ISWT e no EE do Glittre-ADL test foram de moderada a alta magnitude. O Glittre-ADL test apresenta menores respostas metabólicas, cardiovasculares e ventilatórias se comparado ao ISWT.

RESUMEN El presente estudio tuvo el objetivo de evaluar y comparar las respuestas metabólicas, cardiovasculares y ventilatorias de incremental shuttle walk test (ISWT) y de Glittre activities of daily living test (Glittre-ADL test). Es un estudio transversal realizado con individuos sanos. La capacidad funcional (CF) se evaluó utilizando la distancia recorrida y consumo máximo de oxígeno (VO2máx) en el ISWT y el tiempo empleado y VO2 en el Glittre-ADL test. Treinta individuos caminaron 656,67 (IC95%:608,8-704,5) metros en el ISWT y realizaron el Glittre-ADL test en 2,4 (IC95%:2,2-2,6) minutos. El VO2 máx del ISWT fue de 27,8 (IC95%25,6-29,9) versus 22,2 (IC95%20,5-24,1)mL×kg−1×min−1 (p<0,001) en el estado estable (EE) del Glittre-ADL test. Las correlaciones entre la distancia recorrida en el ISWT y el tiempo empleado en el Glittre-ADL test, el VO2máx del ISWT y el VO2 en el EE de Glittre-ADL test y la FC en el máximo del ISWT y en el EE de Glittre-ADL test fueron de moderada a alta magnitud. El Glittre-ADL test presenta respuestas metabólicas, cardiovasculares y ventilatorias más bajas en comparación con el ISWT.

ABSTRACT The aim of the study was to assess and compare the cardiovascular, ventilatory and metabolic responses of the Incremental Shuttle Walk test (ISWT) and Glittre Activities of Daily Living test (Glittre-ADL test). This is a cross-sectional study with individuals. The functional capacity (FC) was evaluated by distance and peak oxygen consumption (VO2peak) in the ISWT and time spent and VO2 in Glittre-ADL test. Thirty individuals went through 656.67 (CI95%:608.8-704.5) meters at the ISWT and performed the Glittre-ADL test in 2.4 (CI95%:2.2-2.6) minutes. The peak VO2 of the ISWT was 27.8 (CI95%25.6-29.9) vs. 22.2 (CI95%20.5-24.1) mL×kg−1×min−1 (p<0.001) in the steady state (SS) of the Glittre-ADL test. Correlations between distance traveled in the ISWT and the time spent in Glittre-ADL test, VO2 peak of ISWT and VO2 in SS of Glittre-ADL test and HR at the ISWT peak and at the Glittre-ADL test SS were moderate to high magnitude. The Glittre-ADL test has lower metabolic, cardiovascular and ventilatory responses compared to ISWT, despite correlations between variables.

Consultoria em amamentação durante a pandemia COVID-19: relato de experiência / Asesoría en lactancia materna durante la pandemia de COVID-19: informe de experiencia / Breastfeeding consultancy during the COVID-19 pandemic: experience report

Objetivo: relatar a experiência de consultoras em amamentação no atendimento às lactantes durante a pandemia de COVID-19. Método: relato de experiência de atendimentos de consultoria em amamentação, nas modalidades online e presencial, na cidade de Fortaleza-CE, Brasil, entre março e abril de 2020. Resultados: diante de sentimentos de medo e ansiedade das lactantes, as consultoras prestaram apoio por meio de aconselhamento, bem como orientações com equipe multiprofissional. Todas as lactantes estavam amamentando e realizando distanciamento social e cuidados de higiene pessoal, sendo reforçado pelas consultoras a importância do acesso às fontes confiáveis de informação. Quanto aos problemas mamários, foram realizadas intervenções preventivas e curativas. Percebeu-se a satisfação das lactantes pelo serviço. Conclusão e implicações para a prática: a consultoria em amamentação constitui-se como um dispositivo agregador na saúde das mulheres que favorece tanto a promoção do aleitamento materno quanto a saúde mental durante a pandemia de Covid-19. Este relato traz direcionamentos para uma prática holística, com vistas à melhoria da qualidade do cuidado prestado, levando-se em consideração os atuais desafios da promoção da saúde diante da pandemia e pode fomentar novas estratégias exitosas

Objective: to report the experience of breastfeeding consultants in caring for breastfeeding women during the Covid-19 pandemic. Method: an experience report of breastfeeding consultancy services, online and face-to-face modalities, in the city of Fortaleza­CE, between March and April 2020. Results: given the mothers' feelings of fear and anxiety, the consultants provided support through counseling, as well as guidance with a multidisciplinary team. All the women were breastfeeding and performing social distancing and personal hygiene care actions, the importance of access to reliable information sources being reinforced by the consultants. As for breast problems, preventive and curative interventions were carried out. The mothers' satisfaction with the consultant service was noticed. Conclusion and implications for practice: breastfeeding consultancy is an aggregating device in women's health that favors both the promotion of breastfeeding and mental health during the Covid-19 pandemic. This report provides directions for a holistic practice, to improve the provided care quality, considering the current challenges of health promotion given the pandemic, and can foster new successful strategies

Objetivo: relatar la experiencia de consultores de lactancia materna en el cuidado de mujeres en periodo de lactancia durante la pandemia de Covid-19. Método: informe de experiencia de los servicios de consultoría en lactancia materna en modalidades en línea y cara a cara, en la ciudad de Fortaleza-CE, entre marzo y abril de 2020. Resultados: ante los sentimientos de miedo y ansiedad de las mujeres lactantes, los consultores proporcionaron apoyo a través de asesoramiento, así como orientación con un equipo multidisciplinario. Todas las mujeres amamantaban y practicaban el distanciamiento social y cuidados de higiene personal, siendo que los consultores reforzaron la importancia del acceso a fuentes confiables de información. En cuanto a los problemas mamarios, se realizaron intervenciones preventivas y curativas. Se notó la satisfacción de las madres lactantes por el servicio. Conclusión e implicaciones para la práctica: la consultoría sobre lactancia materna es un dispositivo agregado en la salud de la mujer que favorece tanto la promoción de la lactancia materna como la salud mental frente a la pandemia de Covid-19. Este estudio proporciona instrucciones para una práctica holística, con miras a mejorar la calidad de la atención brindada, teniendo en cuenta los desafíos actuales de la promoción de la salud frente a la pandemia y puede fomentar nuevas estrategias exitosas

Reverse logistics systems in Brazil: Comparative study and interest of multistakeholders.

Inadequate solid waste management leads to contamination of the soil and surface and underground water; the emergence of antibiotic-resistant superbacteria due to the inadequate disposal of medicines in toilets or in household rubbish; and the emission of persistent organic pollutants, such as flame retardants, present in some electrical/electronic equipment. In Brazil, the principles that underpin the National Solid Waste Policy (PNRS) are consistent with those used by developed countries, namely, the polluter-payer and protector-receiver; cooperation among the different spheres of government, the business sector, and other societal segments; and the shared product life-cycle responsibility (Brasil, 2010a; Novi et al., 2013; Santos et al., 2019). In the latter, selective collection and reverse logistics systems are the primary instruments (Zhang et al., 2011). Thus, the general objective of this work is to compare the technical and economic feasibility of four different sectorial agreements for reverse logistics implemented in Brazil. The specific objective is to describe the possible conflicts of interest of the multistakeholders. The sources of evidence used in this multi-case study were interviews, observations, and documentary research in the period from 2016 to 2018. The first result demonstrated that the economic viability of reverse logistics systems are obtained when the wastes have a commercial value after consumption (packaging and consumer electronics) or when industries organize to charge a fee that covers the costs of recycling and transport (lamps and medicines). The second result shows that there are many stakeholders involved in this process: manufacturers and importers, consumers, private waste and recycling companies, and government represented by municipalities, the Ministry of the Environment (MMA), and state environmental agencies (OEMAs). Each one has different interests and responsibilities: government has to preserve the environment and comply with international agreements on climate change, thus creating laws that ultimately burden companies and cities; profit-maximizing companies evaluate these rules as additional costs in their operations, whereas cities, in which operations are conducted, have low financial and managerial capacity. The stakeholder map reveals that there are overlapping roles among agents, with some being the same for all chains, and the constructed power and level of interest matrix indicates the importance of the process to be conducted by the state to align economic and social-environmental interests.

Diversity of astacin-like metalloproteases identified by transcriptomic analysis in Peruvian Loxosceles laeta spider venom and in vitro activity characterization.

Loxosceles spiders are found in almost all countries of South America. In Peru, Loxosceles laeta species is the main responsible for the accidents caused by poisonous animals, being known as "killer spiders", due to the large number of fatal accidents observed. Astacin-like metalloproteases, named LALPs (Loxosceles astacin-like metalloproteases) are highly expressed in Loxosceles spiders venom gland. These proteases may be involved in hemorrhage and venom spreading, being relevant to the envenoming proccess. Thus, the aim of this work was to analyze Peruvian L. laeta venom gland transcripts using bioinformatics tools, focusing on LALPs. A cDNA library from Peruvian L. laeta venom glands was constructed and sequenced by MiSeq (Illumina) sequencer. After assembly, the resulting sequences were annotated, seeking out for similarity with previously described LALPs. Nine possible LALPs isoforms from Peruvian L. laeta venom were identified and the results were validated by in silico and in vitro experiments. This study contributes to a better understanding of the molecular diversity of Loxosceles venom and provide insights about the action of LALPs.