Phylodynamics of avian influenza A(H5N1) viruses from outbreaks in Brazil.

Our study identified strains of the A/H5N1 virus in analyzed samples of subsistence poultry, wild birds, and mammals, belonging to clade 2.3.4.4b, genotype B3.2, with very high genetic similarity to strains from Chile, Uruguay, and Argentina. This suggests a migratory route for wild birds across the Pacific, explaining the phylogenetic relatedness. The Brazilian samples displayed similarity to strains that had already been previously detected in South America. Phylogeographic analysis suggests transmission of US viruses from Europe and Asia, co-circulating with other lineages in the American continent. As mutations can influence virulence and host specificity, genomic surveillance is essential to detect those changes, especially in critical regions, such as hot spots in the HA, NA, and PB2 sequences. Mutations in the PB2 gene (D701N and Q591K) associated with adaptation and transmission in mammals were detected suggesting a potential zoonotic risk. Nonetheless, resistance to neuraminidase inhibitors (NAIs) was not identified, however, continued surveillance is crucial to detect potential resistance. Our study also mapped the spread of the virus in the Southern hemisphere, identifying possible entry routes and highlighting the importance of surveillance to prevent outbreaks and protect both human and animal populations.

[Legal abortion in cases of sexual violence pregnancy: perceptions and experiences of obstetricians]. / O aborto legal em casos de gravidez decorrente de violência sexual: percepções e vivências de médicas e médicos obstetras.

This study aimed to analyze the perceptions of obstetricians and gynecology-obstetrics residents at a federal school maternity hospital regarding legal abortion in cases of sexual violence pregnancy, understand their motivations, strengths, and feelings, and identify their experience with this topic. The first stage consisted of answering a self-administered questionnaire. The selection criteria were: obstetricians linked to the obstetric center, director of the medical division, and residents of the institution's obstetrics-gynecology program. In total, 36 questionnaires were answered and returned. The second stage corresponded to an interview using a saturation sampling criterion. Six physicians were interviewed. The interviews were evaluated using a thematic content analysis. The questionnaires showed that all participants had already provided care to women in situations of sexual violence and that most of them had already participated in a legal abortion procedure in these cases. The interviews highlighted the dilemmas faced by professionals in providing care to these women and the lack of professional training to handle these cases. The speeches of women were sometimes seen as an object of suspicion regarding the veracity of sexual violence, and sometimes as an object that caused professionals to feel emotionally affected while listening to them, allowing professionals to approach the victims and offer more humanized care. The results pointed to the importance of addressing this topic in the areas of health and providing training beyond the technical-scientific focus in order to support the development of new care strategies.

Este estudo objetivou analisar as percepções de obstetras e residentes de ginecologia-obstetrícia, atuantes numa maternidade escola federal, sobre o aborto legal em casos de gravidez decorrente de violência sexual, desvelando suas motivações, resistências e sentimentos, e identificando suas experiências com o tema. A primeira etapa correspondeu ao preenchimento de um questionário autoaplicável. Os critérios de seleção foram: obstetras vinculados ao centro obstétrico; diretor da divisão médica; e residentes do programa de ginecologia-obstetrícia da instituição. Obtiveram-se 36 questionários respondidos. A segunda etapa correspondeu à realização de uma entrevista, tendo sido utilizado o critério de amostragem por saturação e foram entrevistados seis médicos. As entrevistas foram analisadas pelo método de análise de conteúdo, na modalidade temática. Os questionários retrataram que todos os participantes já haviam prestado assistência a mulheres em situação de violência sexual e que a maioria já havia participado da realização de um aborto legal. As entrevistas evidenciaram os dilemas enfrentados pelos profissionais na assistência a esses casos e a escassez da formação profissional em relação à temática. A palavra da mulher foi tida ora como objeto de suspeição em relação à veracidade do estupro, ora como capaz de suscitar afetação das profissionais em suas escutas, o que possibilitou que essas se aproximassem das vítimas e ofertassem uma assistência mais humanizada. Os resultados apontaram para a importância da temática ser abordada nos campos da saúde e da formação para além do enfoque técnico-científico, visando produzir novas estratégias de cuidado.

El estudio tuvo como objetivo analizar las percepciones de obstetras y residentes de gineco-obstetricia de una maternidad escolar federal sobre el aborto legal en casos de embarazo resultante de violencia sexual, revelar sus motivaciones, resistencias y sentimientos, e identificar sus experiencias con el tema. La primera etapa consistió en completar un cuestionario autoadministrado. Los criterios de selección fueron los siguientes: obstetras vinculados al centro obstétrico; director de la división médica; y residentes del programa de gineco-obstetricia de la institución. Se obtuvieron 36 cuestionarios cumplimentados. La segunda etapa consistió en una entrevista, utilizando un criterio de muestreo por saturación. Se entrevistó a 6 médicos. Las entrevistas se analizaron mediante el método de análisis de contenido, en la modalidad temática. Los cuestionarios mostraron que todos los participantes ya habían brindado asistencia a mujeres en situación de violencia sexual y que la mayoría ya había participado en la realización de un aborto legal en estos casos. Las entrevistas pusieron de manifiesto los dilemas que enfrentan los profesionales en la asistencia a estos casos y la escasa formación profesional con relación al tema. La palabra de la mujer fue vista a veces como objeto de sospecha con respecto a la veracidad de la violación, y a veces como un objeto capaz de suscitar la afectación de las profesionales en sus escuchas, lo que les permitió acercarse a las víctimas y ofrecer una asistencia más humanizada. Los resultados señalaron la importancia de que la temática sea abordada en los campos de la salud y de la formación más allá del enfoque técnico-científico, con el objetivo de producir nuevas estrategias de cuidado.

Phylogeny, envenomation syndrome, and membrane permeabilising venom produced by Australia's electric caterpillar Comana monomorpha.

Zygaenoidea is a superfamily of lepidopterans containing many venomous species, including the Limacodidae (nettle caterpillars) and Megalopygidae (asp caterpillars). Venom proteomes have been recently documented for several species from each of these families, but further data are required to understand the evolution of venom in Zygaenoidea. In this study, we examined the 'electric' caterpillar from North-Eastern Australia, a limacodid caterpillar densely covered in venomous spines. We used DNA barcoding to identify this caterpillar as the larva of the moth Comana monomorpha (Turner, 1904). We report the clinical symptoms of C. monomorpha envenomation, which include acute pain, and erythema and oedema lasting for more than a week. Combining transcriptomics of venom spines with proteomics of venom harvested from the spine tips revealed a venom markedly different in composition from previously examined limacodid venoms that are rich in peptides. In contrast, the venom of C. monomorpha is rich in aerolysin-like proteins similar to those found in venoms of asp caterpillars (Megalopygidae). Consistent with this composition, the venom potently permeabilises sensory neurons and human neuroblastoma cells. This study highlights the diversity of venom composition in Limacodidae.

Sexual and reproductive health: analysis of the content published in two Brazilian scientific nursing events.

OBJECTIVE: To analyze the content published in the Proceedings of the 71st Brazilian Nursing Congress and the 20th National Research Seminar in the field of sexual and reproductive health. METHOD: Qualitative, descriptive study, whose data sources were the annalsof two 2019 events that were related to Sexual and Reproductive Health. Content analysis and lexical typology were carried out with the aid of the IRAMUTEq® software. RESULTS: In a universe of 3,433 abstracts, 603 were analyzed and showed a higher prevalence of qualitative studies and experience reports, focusing on the maternal-child area and on the pathologizing and medicalizing processes of women's health, with incipient use of software in the analysis. However, there were signs of health promotion and humanized care. CONCLUSION: The dissemination of studies in the field of sexual and Reproductive Health seems to be in a conflict between accommodation and resistance, at the same time that it maintains old imperatives of the hegemonic domain, seeking to overcome them with new methodologies and care based on integrality and equity.

Tissue damaging toxins in snake venoms: mechanisms of action, pathophysiology and treatment strategies.

Snakebite envenoming is an important public health issue responsible for mortality and severe morbidity. Where mortality is mainly caused by venom toxins that induce cardiovascular disturbances, neurotoxicity, and acute kidney injury, morbidity is caused by toxins that directly or indirectly destroy cells and degrade the extracellular matrix. These are referred to as 'tissue-damaging toxins' and have previously been classified in various ways, most of which are based on the tissues being affected (e.g., cardiotoxins, myotoxins). This categorisation, however, is primarily phenomenological and not mechanistic. In this review, we propose an alternative way of classifying cytotoxins based on their mechanistic effects rather than using a description that is organ- or tissue-based. The mechanisms of toxin-induced tissue damage and their clinical implications are discussed. This review contributes to our understanding of fundamental biological processes associated with snakebite envenoming, which may pave the way for a knowledge-based search for novel therapeutic options.

Inhibition of N-type calcium channels by phenoxyaniline and sulfonamide analogues.

Building on previous investigations, structural modifications to the neuronal calcium ion channel blocker MONIRO-1 and related compounds were conducted that included replacement of the amide linker with an aniline and isosteric sulfonamide moiety, and the previously used strategy of substitution of the guanidinium group with less hydrophilic amine functionalities. A comprehensive SAR study revealed a number of phenoxyaniline and sulfonamide compounds that were more potent or had similar potency for the CaV2.2 and CaV3.2 channel compared to MONIRO-1 when evaluated in a FLIPR-based intracellular calcium response assay. Cytotoxicity investigations indicated that the sulfonamide analogues were well tolerated by Cos-7 cells at dosages required to inhibit both calcium ion channels. The sulfonamide derivatives were the most promising CaV2.2 inhibitors developed by us to date due, possessing high stability in plasma, low toxicity (estimated therapeutic index > 10), favourable CNS MPO scores (4.0-4.4) and high potency and selectivity, thereby, making this class of compounds suitable candidates for future in vivo studies.

High-content fluorescence bioassay investigates pore formation, ion channel modulation and cell membrane lysis induced by venoms.

Venoms comprise highly sophisticated bioactive molecules modulating ion channels, receptors, coagulation factors, and the cellular membranes. This array of targets and bioactivities requires advanced high-content bioassays to facilitate the development of novel envenomation treatments and biotechnological and pharmacological agents. In response to the existing gap in venom research, we developed a cutting-edge fluorescence-based high-throughput and high-content cellular assay. This assay enables the simultaneous identification of prevalent cellular activities induced by venoms such as membrane lysis, pore formation, and ion channel modulation. By integrating intracellular calcium with extracellular nucleic acid measurements, we have successfully distinguished these venom mechanisms within a single cellular assay. Our high-content bioassay was applied across three cell types exposed to venom components representing lytic, ion pore-forming or ion channel modulator toxins. Beyond unveiling distinct profiles for these action mechanisms, we found that the pore-forming latrotoxin α-Lt1a prefers human neuroblastoma to kidney cells and cardiomyocytes, while the lytic bee peptide melittin is not selective. Furthermore, evaluation of snake venoms showed that Elapid species induced rapid membrane lysis, while Viper species showed variable to no activity on neuroblastoma cells. These findings underscore the ability of our high-content bioassay to discriminate between clades and interspecific traits, aligning with clinical observations at venom level, beyond discriminating among ion pore-forming, membrane lysis and ion channel modulation. We hope our research will expedite the comprehension of venom biology and the diversity of toxins that elicit cytotoxic, cardiotoxic and neurotoxic effects, and assist in identifying venom components that hold the potential to benefit humankind.

Detection of Trichomonas vaginalis by Allplex™ STI Essential Assay (Seegene) in clinical samples from the Brazilian public health system users.

The failures in Trichomonas vaginalis (TV) infection diagnosis leave more than half of cases unidentified. In this report, urine and vaginal discharge samples were analyzed by wet mount, culture examination, and real-time PCR by Allplex™ (Seegene®) kit, in a population assisted by the Brazilian Public Health System. From 747 samples, 2.81% were positive for TV in wet mount and culture, and 3.88% by Allplex™. Samples kept at - 80 ºC for 22 months did not impair the PCR technique. The sensitivity for wet mount, culture, and Allplex™ was 72, 100, and 100%, respectively. Allplex™ technique showed highest detection of TV.

O aborto legal em casos de gravidez decorrente de violência sexual: percepções e vivências de médicas e médicos obstetras / Legal abortion in cases of sexual violence pregnancy: perceptions and experiences of obstetricians / El aborto legal en casos de embarazo resultante de violencia sexual: percepciones y experiencias de médicas y médicos obstetras

Resumo: Este estudo objetivou analisar as percepções de obstetras e residentes de ginecologia-obstetrícia, atuantes numa maternidade escola federal, sobre o aborto legal em casos de gravidez decorrente de violência sexual, desvelando suas motivações, resistências e sentimentos, e identificando suas experiências com o tema. A primeira etapa correspondeu ao preenchimento de um questionário autoaplicável. Os critérios de seleção foram: obstetras vinculados ao centro obstétrico; diretor da divisão médica; e residentes do programa de ginecologia-obstetrícia da instituição. Obtiveram-se 36 questionários respondidos. A segunda etapa correspondeu à realização de uma entrevista, tendo sido utilizado o critério de amostragem por saturação e foram entrevistados seis médicos. As entrevistas foram analisadas pelo método de análise de conteúdo, na modalidade temática. Os questionários retrataram que todos os participantes já haviam prestado assistência a mulheres em situação de violência sexual e que a maioria já havia participado da realização de um aborto legal. As entrevistas evidenciaram os dilemas enfrentados pelos profissionais na assistência a esses casos e a escassez da formação profissional em relação à temática. A palavra da mulher foi tida ora como objeto de suspeição em relação à veracidade do estupro, ora como capaz de suscitar afetação das profissionais em suas escutas, o que possibilitou que essas se aproximassem das vítimas e ofertassem uma assistência mais humanizada. Os resultados apontaram para a importância da temática ser abordada nos campos da saúde e da formação para além do enfoque técnico-científico, visando produzir novas estratégias de cuidado.

Abstract: This study aimed to analyze the perceptions of obstetricians and gynecology-obstetrics residents at a federal school maternity hospital regarding legal abortion in cases of sexual violence pregnancy, understand their motivations, strengths, and feelings, and identify their experience with this topic. The first stage consisted of answering a self-administered questionnaire. The selection criteria were: obstetricians linked to the obstetric center, director of the medical division, and residents of the institution's obstetrics-gynecology program. In total, 36 questionnaires were answered and returned. The second stage corresponded to an interview using a saturation sampling criterion. Six physicians were interviewed. The interviews were evaluated using a thematic content analysis. The questionnaires showed that all participants had already provided care to women in situations of sexual violence and that most of them had already participated in a legal abortion procedure in these cases. The interviews highlighted the dilemmas faced by professionals in providing care to these women and the lack of professional training to handle these cases. The speeches of women were sometimes seen as an object of suspicion regarding the veracity of sexual violence, and sometimes as an object that caused professionals to feel emotionally affected while listening to them, allowing professionals to approach the victims and offer more humanized care. The results pointed to the importance of addressing this topic in the areas of health and providing training beyond the technical-scientific focus in order to support the development of new care strategies.

Resumen: El estudio tuvo como objetivo analizar las percepciones de obstetras y residentes de gineco-obstetricia de una maternidad escolar federal sobre el aborto legal en casos de embarazo resultante de violencia sexual, revelar sus motivaciones, resistencias y sentimientos, e identificar sus experiencias con el tema. La primera etapa consistió en completar un cuestionario autoadministrado. Los criterios de selección fueron los siguientes: obstetras vinculados al centro obstétrico; director de la división médica; y residentes del programa de gineco-obstetricia de la institución. Se obtuvieron 36 cuestionarios cumplimentados. La segunda etapa consistió en una entrevista, utilizando un criterio de muestreo por saturación. Se entrevistó a 6 médicos. Las entrevistas se analizaron mediante el método de análisis de contenido, en la modalidad temática. Los cuestionarios mostraron que todos los participantes ya habían brindado asistencia a mujeres en situación de violencia sexual y que la mayoría ya había participado en la realización de un aborto legal en estos casos. Las entrevistas pusieron de manifiesto los dilemas que enfrentan los profesionales en la asistencia a estos casos y la escasa formación profesional con relación al tema. La palabra de la mujer fue vista a veces como objeto de sospecha con respecto a la veracidad de la violación, y a veces como un objeto capaz de suscitar la afectación de las profesionales en sus escuchas, lo que les permitió acercarse a las víctimas y ofrecer una asistencia más humanizada. Los resultados señalaron la importancia de que la temática sea abordada en los campos de la salud y de la formación más allá del enfoque técnico-científico, con el objetivo de producir nuevas estrategias de cuidado.

Racial inequality in health care of adults hospitalized with COVID-19. / Desigualdade racial na assistência à saúde do adulto internado por COVID-19.

The objective was to analyze the association of race/skin color in health care, in adults hospitalized with severe acute respiratory syndrome (SARS)/COVID-19, between March 2020 and September 2022, with Brazil as the unit of analysis. This is a cross-sectional study that used the Influenza Epidemiological Surveillance Information System (SIVEP-Gripe) database and had a population composed of adults (≥ 18 years) and the final classification was SARS by COVID-19 or unspecified SARS. The direct effect of skin color on in-hospital mortality was estimated through logistic regression adjusted for age, gender, schooling level, health care system and period, stratified by vaccination status. This same model was also used to assess the effect of skin color on the variables related to access to health care services: intensive care unit (ICU), tomography, chest X-ray and ventilatory support. The results show that black, brown and indigenous people died more, regardless the schooling level and number of comorbidities, with 23%, 32% and 80% higher chances of death, respectively, when submitted to ventilatory support. Racial differences were observed in the use of health care services and in outcomes of death from COVID-19 or unspecified SARS, in which ethnic minorities had higher in-hospital mortality and lower use of hospital resources. These results suggest that black and indigenous populations have severe disadvantages compared to the white population, facing barriers to access health care services in the context of the COVID-19 pandemic.

O objetivo deste artigo foi analisar a associação entre raça/cor e assistência à saúde, em adultos hospitalizados pela síndrome respiratória aguda grave (SRAG)/COVID-19 no Brasil, entre março de 2020 e setembro de 2022. Trata-se de estudo transversal, que utilizou o banco de dados do Sistema de Informação de Vigilância Epidemiológica da Gripe (SIVEP-Gripe) e contou com uma população composta por adultos (≥ 18 anos). A classificação final foi SRAG por COVID-19 ou SRAG não especificada. O efeito direto do aspecto cor na mortalidade intra-hospitalar foi estimado por meio de regressão logística ajustada por idade, sexo, escolaridade, sistema de saúde e período, estratificado por situação vacinal. Esse mesmo modelo foi utilizado também para avaliar o efeito do quesito cor nas variáveis de acesso aos serviços de saúde: unidade de terapia intensiva (UTI), tomografia, radiografia de tórax e suporte ventilatório. Os resultados evidenciam que pretos, pardos e indígenas morreram mais, independentemente do grau de escolaridade e da quantidade de comorbidades, com maiores chances de óbito em 23%, 32% e 80%, respectivamente, ao serem submetidos ao suporte ventilatório. Foram observadas diferenças raciais no uso de serviços de saúde e nos desfechos de morte por COVID-19 ou SRAG não especificada, em que minorias étnicas tiveram maiores taxas de mortalidade intra-hospitalar e os recursos hospitalares foram utilizados com menos frequência. Tais resultados sugerem que as populações negra e indígena têm severas desvantagens em relação à branca, enfrentando barreiras de acesso aos serviços de saúde no contexto da pandemia de COVID-19.

Se pretende analizar la asociación entre raza/color en la atención de la salud de adultos hospitalizados por síndrome respiratorio agudo severo (SARS)/COVID-19 en el período entre marzo de 2020 y septiembre de 2022, tomando Brasil como unidad de análisis. Se trata de un estudio transversal, en que se utilizó la base de datos del Sistema de Información de Vigilancia Epidemiológica de la Gripe (SIVEP-Gripe) y tuvo una muestra compuesta por adultos (≥ 18 años) y la clasificación final fue SARS por COVID-19 o SARS no especificado. El efecto directo de la pregunta color sobre la mortalidad intrahospitalaria se estimó mediante la regresión logística ajustada según edad, sexo, nivel de estudios, sistema de salud y período, estratificada por estado de vacunación. Este modelo también se utilizó para evaluar el efecto de la pregunta color en las variables de acceso a los servicios de salud: unidades de cuidado intensivo (UCI), tomografía, radiografía de tórax y soporte ventilatorio. Los resultados muestran que negros, pardos e indígenas murieron más, independientemente del nivel de estudios y el número de comorbilidades, y cuando fueron sometidos a soporte ventilatorio, tuvieron mayores probabilidades de muerte con el 23%, 32% y 80%, respectivamente. Se observaron diferencias raciales en el uso de los servicios de salud y los resultados de muerte por COVID-19 o SARS no especificado, en los que las minorías étnicas tuvieron una mayor mortalidad intrahospitalaria y los recursos hospitalarios se utilizaron con menos frecuencia. Estos resultados evidencian que las poblaciones negra e indígena tienen graves desventajas en comparación con la población blanca, afrontando dificultades de acceso a los servicios de salud en el contexto de la pandemia del COVID-19.

Structure-function and rational design of a spider toxin Ssp1a at human voltage-gated sodium channel subtypes.

The structure-function and optimization studies of NaV-inhibiting spider toxins have focused on developing selective inhibitors for peripheral pain-sensing NaV1.7. With several NaV subtypes emerging as potential therapeutic targets, structure-function analysis of NaV-inhibiting spider toxins at such subtypes is warranted. Using the recently discovered spider toxin Ssp1a, this study extends the structure-function relationships of NaV-inhibiting spider toxins beyond NaV1.7 to include the epilepsy target NaV1.2 and the pain target NaV1.3. Based on these results and docking studies, we designed analogues for improved potency and/or subtype-selectivity, with S7R-E18K-rSsp1a and N14D-P27R-rSsp1a identified as promising leads. S7R-E18K-rSsp1a increased the rSsp1a potency at these three NaV subtypes, especially at NaV1.3 (∼10-fold), while N14D-P27R-rSsp1a enhanced NaV1.2/1.7 selectivity over NaV1.3. This study highlights the challenge of developing subtype-selective spider toxin inhibitors across multiple NaV subtypes that might offer a more effective therapeutic approach. The findings of this study provide a basis for further rational design of Ssp1a and related NaSpTx1 homologs targeting NaV1.2, NaV1.3 and/or NaV1.7 as research tools and therapeutic leads.

First report and genetic characterization of the highly pathogenic avian influenza A(H5N1) virus in Cabot's tern (Thalasseus acuflavidus), Brazil.

In 2021, the H5N1 virus lineage 2.3.4.4b spread to the Americas, causing high mortality in wild and domestic avian populations. South American countries along the Pacific migratory route have reported wild bird deaths due to A/H5Nx virus since October 2022. However, limited genomic data resulted in no cases reported in Brazil until May 2023. Brazil reported its first case of highly pathogenic avian influenza virus (HPAI A/H5N1) in May 2023. The virus was detected in Cabot's tern specimen in Marataízes, Espírito Santo. Cases were also found in backyard poultry and other wild birds, but no human or commercial poultry cases occurred. HPAI poses risks to the poultry industry, food security, and public health. Researchers used next-gen sequencing and phylogenetic analysis to study the Brazilian sample. It confirmed its affiliation with the 2.3.4.4b clade and proximity to sequences from Chile and Peru. This sheds light on the spread and evolution of HPAI A/H5N1 in the Americas, emphasizing continuous monitoring to mitigate risks for both avian and human populations. Understanding the virus's genetics and transmission allows implementing effective control measures to protect public health and the poultry industry.

Antimicrobial and Antibiofilm Activities of Copper(II)-1,10-phenanthroline-5,6-pione Against Commensal Bacteria and Fungi Responsible for Vaginal Microbiota Dysbiosis.

The disbalance of vaginal eubiotic microbiota can lead to overgrowth of Candida species and bacteria responsible for aerobic vaginitis, activating inflammatory pathways. The presence of Trichomonas vaginalis, a sexually transmitted protozoan pathogen, can be a predisposing factor for disordering the growth of bacterial/fungal pathogenic species due to the increase in pH and reduction of eubiotic microbiota. Herein, we evaluated the effects of the potent trichomonacidal compound, copper(II)-1,10-phenanthroline-5,6-dione (Cu-phendione), against pathogens responsible for candidiasis and aerobic vaginitis. Cu-phendione showed antimicrobial activity against Candida albicans, non-albicans Candida species (C. glabrata, C. krusei, C. parapsilosis, and C. tropicalis) and Gram-negative (Escherichia coli) and Gram-positive (Staphylococcus aureus, Enterococcus faecalis, and Streptococcus agalactiae) bacteria. Moreover, Cu-phendione was able to interfere with the fungal biofilm formation. These results highlight the antimicrobial potential of Cu-phendione against bacterial and fungal strains of vaginitis-causing infectious agents.

From squid giant axon to automated patch-clamp: electrophysiology in venom and antivenom research.

Ion channels play a crucial role in diverse physiological processes, including neurotransmission and muscle contraction. Venomous creatures exploit the vital function of ion channels by producing toxins in their venoms that specifically target these ion channels to facilitate prey capture upon a bite or a sting. Envenoming can therefore lead to ion channel dysregulation, which for humans can result in severe medical complications that often necessitate interventions such as antivenom administration. Conversely, the discovery of highly potent and selective venom toxins with the capability of distinguishing between different isoforms and subtypes of ion channels has led to the development of beneficial therapeutics that are now in the clinic. This review encompasses the historical evolution of electrophysiology methodologies, highlighting their contributions to venom and antivenom research, including venom-based drug discovery and evaluation of antivenom efficacy. By discussing the applications and advancements in patch-clamp techniques, this review underscores the profound impact of electrophysiology in unravelling the intricate interplay between ion channels and venom toxins, ultimately leading to the development of drugs for envenoming and ion channel-related pathologies.

Genipin crosslinked porous chitosan beads as robust supports for ß-galactosidase immobilization: Characterization, stability, and bioprocessing potential.

This study aimed to modify the porosity of chitosan beads using Na2CO3 as a porogen agent and to crosslink them with genipin for the immobilization of ß-galactosidase from Aspergillus oryzae. Immobilization was performed under four different pH conditions (4.5, 6.0, 7.5, and 9.0), resulting in biocatalysts named B4, B6, B7, and B9, respectively. The immobilized enzymes were characterized for immobilization parameters and stability, including thermal, pH, storage, and operational stability. The optimal conditions for the support were determined as 50 mM Na2CO3. The biocatalyst exhibited nearly 100 % retention of initial activity after 5 h of incubation at different pH conditions and showed improved thermal stability compared to the free enzyme across all pH conditions. After 50 cycles of lactose hydrolysis, all biocatalysts retained at least 71 % of their initial activity, with B6 retaining nearly 100 %. Scanning electron microscopy revealed structural modifications, particularly in B4, leading to weakened support structure after reuse. Continuous lactose hydrolysis showed increased productivity from 41.3 to 48.1 g L-1 h-1 for B6, with 78.1 % retention of initial capacity. All biocatalysts retained >95 % activity when stored at 4 °C for 20 weeks, highlighting their suitability for enzyme immobilization in continuous and discontinuous bioprocesses.

Horizontal gene transfer underlies the painful stings of asp caterpillars (Lepidoptera: Megalopygidae).

Larvae of the genus Megalopyge (Lepidoptera: Zygaenoidea: Megalopygidae), known as asp or puss caterpillars, produce defensive venoms that cause severe pain. Here, we present the anatomy, chemistry, and mode of action of the venom systems of caterpillars of two megalopygid species, the Southern flannel moth Megalopyge opercularis and the black-waved flannel moth Megalopyge crispata. We show that megalopygid venom is produced in secretory cells that lie beneath the cuticle and are connected to the venom spines by canals. Megalopygid venoms consist of large aerolysin-like pore-forming toxins, which we have named megalysins, and a small number of peptides. The venom system differs markedly from those of previously studied venomous zygaenoids of the family Limacodidae, suggestive of an independent origin. Megalopygid venom potently activates mammalian sensory neurons via membrane permeabilization and induces sustained spontaneous pain behavior and paw swelling in mice. These bioactivities are ablated by treatment with heat, organic solvents, or proteases, indicating that they are mediated by larger proteins such as the megalysins. We show that the megalysins were recruited as venom toxins in the Megalopygidae following horizontal transfer of genes from bacteria to the ancestors of ditrysian Lepidoptera. Megalopygids have recruited aerolysin-like proteins as venom toxins convergently with centipedes, cnidarians, and fish. This study highlights the role of horizontal gene transfer in venom evolution.

Development and application of proficiency testing (PT) to evaluate the diagnostic capacity of SARS-CoV-2 by RT-qPCR - A practical and metrological approach.

It is necessary to use quality tools to evaluate the diagnostic capacity of laboratories, such as implementing a proficiency testing (PT) program. The goal of this work is to develop and apply a PT protocol to assess the diagnostic capacity of SARS-CoV-2 through the RT-PCR method, based on appropriate metrological tools. A 5-item test panel containing items with different dilutions of SARS-CoV-2, including negative controls, was developed to perform this PT with the application of different performance assessment tools to score and differentiate performance between laboratories, according to Table 2. Based on the participants' total qualitative result, 95% of the negative samples and 73% of the positive samples were correctly identified by the laboratories. The results obtained were compared e validate the systematics of the PT developed, so that it can be implemented and used to monitor and improve the diagnostic capacity of SARS-CoV-2, also helping to improve the quality of these results.

The prevalence and perinatal repercussions of preeclampsia after the implementation of a prophylaxis protocol with aspirin.

OBJECTIVES: To evaluate the prevalence and perinatal repercussions of preeclampsia (PE) after the implementation of a prophylaxis protocol with aspirin in singleton pregnancy at Maternity School of Federal University of Rio de Janeiro, Rio de Janeiro, Brazil (2015-2106). METHODOLOGY: PE prevalence according to gestational age (GA) and the prevalence ratio (PR) between PE and prematurity, small for gestational age (SGA), and fetal death were calculated in patients assisted during 2015 and 2016. RESULTS: PE occurred in 373(10.75%) of 3468 investigated cases, where PE < 37 weeks was of 2.79% and PE greater than 37 weeks was of 7.95%. A total of 413 (11.9%) prematurity cases, 320 SGA (9.22%), and 50 fetal deaths (1.44%) occurred. In the PE group, 97 premature newborns (PR 0.90) and 51 SGA (PR 1.16) were born, and two fetal deaths occurred (PR 7.46). Concerning PE < 37 weeks, 27 SGA cases (PR 1.42) and two fetal deaths (PR 2.62) were observed. Regarding PE greater than 37 weeks, 24 SGA (PR 1.09) were born, and no fetal deaths were observed. Our findings were compared to previously published results. CONCLUSIONS: PE was significantly associated with SGA newborns, especially premature PE. Prescribing aspirin for PE prophylaxis based only on clinical risk factors in a real-life scenario does not appear to be effective but resulted in a PE screening and prophylaxis protocol review and update at ME/UFRJ.

Peptidases Are Potential Targets of Copper(II)-1,10-Phenanthroline-5,6-dione Complex, a Promising and Potent New Drug against Trichomonas vaginalis.

Trichomonas vaginalis is responsible for 156 million new cases per year worldwide. When present asymptomatically, the parasite can lead to serious complications, such as development of cervical and prostate cancer. As infection increases the acquisition and transmission of HIV, the control of trichomoniasis represents an important niche for the discovery and development of new antiparasitic molecules. This urogenital parasite synthesizes several molecules that allow the establishment and pathogenesis of infection. Among them, peptidases occupy key roles as virulence factors, and the inhibition of these enzymes has become an important mechanism for modulating pathogenesis. Based on these premises, our group recently reported the potent anti-T. vaginalis action of the metal-based complex [Cu(phendione)3](ClO4)2.4H2O (Cu-phendione). In the present study, we evaluated the influence of Cu-phendione on the modulation of proteolytic activities produced by T. vaginalis by biochemical and molecular approaches. Cu-phendione showed strong inhibitory potential against T. vaginalis peptidases, especially cysteine- and metallo-type peptidases. The latter revealed a more prominent effect at both the post-transcriptional and post-translational levels. Molecular Docking analysis confirmed the interaction of Cu-phendione, with high binding energy (-9.7 and -10.7 kcal·mol-1, respectively) at the active site of both TvMP50 and TvGP63 metallopeptidases. In addition, Cu-phendione significantly reduced trophozoite-mediated cytolysis in human vaginal (HMVII) and monkey kidney (VERO) epithelial cell lineages. These results highlight the antiparasitic potential of Cu-phendione by interaction with important T. vaginalis virulence factors.