Natural products as a control measure of the Achatina fulica (Gastropoda: Achatinidae) / Produtos naturais como medida de controle do molusco Achatina fulica (Gastropoda: Achatinidae)

Achatina fulica is a terrestrial mollusk known as the giant African snail that is related to environmental, economic, urban, and public health problems. As control measures for this mollusk, cooking salt (NaCl) and calcium oxide (CaO) are used, and baits are composed of metaldehyde. However, these measures have environmental toxicity and impact the soil. In this way, natural products have been tested on this mollusk to discover and develop a substance to combat this urban and agricultural pest. This article aims to evaluate studies involving natural products to control the population of Achatina fulica. Articles and works published in books were included in the present work. A total of 1,103 works were found during the search. Of these, 14 works met the objective of these review and were included in this article. The tests do not possess methodological standardization, do not have a maximum concentration to be considered active, or a maximum exposure time. A lack of standardization in the methodology of tests on A. fulica was observed. The performance of tests on other life stages of the mollusk, as well as tests that analyze other parameters, are essential. Only one article analyzed presented phytochemical analysis. No ecotoxicity tests were reported either. Some extracts showed promising results, highlighting the aqueous extract of Capsicum frutescens. More studies investigating the molluscicidal activity of natural products on A. fulica are needed. It is very relevant that the new studies present a phytochemical analysis of the tested extracts, as well as ecotoxicity studies.

Achatina fulica é um molusco terrestre conhecido como caramujo gigante africano que está relacionado a problemas ambientais, econômicos, urbanos e de saúde pública. Como medidas de controle para esse molusco, são utilizados sal de cozinha (NaCl) e óxido de cálcio (CaO), e as iscas são compostas de metaldeído. No entanto, essas medidas têm toxicidade ambiental e impactam o solo. Desta forma, produtos naturais foram testados neste molusco para descobrir e desenvolver uma substância para combater esta praga urbana e agrícola. Este artigo tem como objetivo avaliar estudos envolvendo produtos naturais para controle da população de Achatina fulica. Artigos e trabalhos publicados em livros foram incluídos no presente trabalho. Um total de 1.103 trabalhos foram encontrados durante a pesquisa. Destes, 14 trabalhos atendiam ao objetivo desta revisão e foram incluídos neste artigo. Os testes não possuem padronização metodológica, não possuem concentração máxima para serem considerados ativos ou tempo máximo de exposição. Observou-se uma falta de padronização na metodologia de testes em A. fulica. A realização de testes em outras fases da vida do molusco, bem como testes que analisem outros parâmetros, são essenciais. Apenas um artigo analisado apresentou análise fitoquímica. Também não foram relatados testes de ecotoxicidade. Alguns extratos apresentaram resultados promissores, com destaque para o extrato aquoso de Capsicum frutescens. Mais estudos investigando a atividade molusquicida de produtos naturais sobre A. fulica são necessários. É muito relevante que os novos estudos apresentem uma análise fitoquímica dos extratos testados, bem como estudos de ecotoxicidade.

Bioactivity of substances isolated from natural products on mollusks Biomphalaria glabrata (Say, 1818) (Planorbidae): a review.

Schistosomiasis is a neglected tropical disease caused by parasitic worms of several species of the genus Schistosoma. Transmission occurs by parasitic larvae that stay in freshwater snails of the genus Biomphalaria. Thus, the search for new products that are biodegradable has increased the interest in products of plant origin. The aim of this article is to review the isolated substances from natural products that showed molluscicidal activity against the species Biomphalaria glabrata in order to reevaluate the most promising prototypes and update the progress of research to obtain a new molluscicide. We perform searches using scientific databases, such as Scientific Electronic Library Online (SciELO), Google schoolar, PUBMED, Web of Science and Latin American and Caribbean Literature on Health Sciences (LILACS). From 2000 to 2022, using the keywords "isolated substances", "molluscicidal activity" and "Biomphalaria glabrata". In the present study, it was possible to observe 19 promising molluscicidal molecules with a lethal concentration below 20 µg/mL. Of these promising isolates, only 5 isolates had the CL90 calculated and within the value recommended by WHO: Benzoic acid, 2',4',6'-Trihydroxydihydrochalcone, Divaricatic acid, Piplartine and 2-hydroxy-1,4-naphthoquinone (Lapachol). We conclude that beyond a few results in the area, the researches don't follow the methodological pattern (exposure time and measure units, toxicity test), in this way, as they don't follow a pattern on the result's exposure (LC), not following, in sum, the recommended by WHO.

Bioactivity of substances isolated from natural products on mollusks Biomphalaria glabrata (Say, 1818) (Planorbidae): a review / Bioatividade de substâncias isoladas de produtos naturais em moluscos Biomphalaria glabrata (Say, 1818) (Planorbidae): uma revisão

Schistosomiasis is a neglected tropical disease caused by parasitic worms of several species of the genus Schistosoma. Transmission occurs by parasitic larvae that stay in freshwater snails of the genus Biomphalaria. Thus, the search for new products that are biodegradable has increased the interest in products of plant origin. The aim of this article is to review the isolated substances from natural products that showed molluscicidal activity against the species Biomphalaria glabrata in order to reevaluate the most promising prototypes and update the progress of research to obtain a new molluscicide. We perform searches using scientific databases, such as Scientific Electronic Library Online (SciELO), Google schoolar, PUBMED, Web of Science and Latin American and Caribbean Literature on Health Sciences (LILACS). From 2000 to 2022, using the keywords "isolated substances", "molluscicidal activity" and "Biomphalaria glabrata". In the present study, it was possible to observe 19 promising molluscicidal molecules with a lethal concentration below 20 µg/mL. Of these promising isolates, only 5 isolates had the CL90 calculated and within the value recommended by WHO: Benzoic acid, 2',4',6'-Trihydroxydihydrochalcone, Divaricatic acid, Piplartine and 2-hydroxy-1,4-naphthoquinone (Lapachol). We conclude that beyond a few results in the area, the researches don't follow the methodological pattern (exposure time and measure units, toxicity test), in this way, as they don't follow a pattern on the result's exposure (LC), not following, in sum, the recommended by WHO.

A esquistossomose é uma doença tropical negligenciada causada por vermes parasitas de várias espécies do gênero Schistosoma. A transmissão ocorre por larvas parasitas que ficam em caramujos de água doce do gênero Biomphalaria. Assim, a busca por novos produtos biodegradáveis ​​tem aumentado o interesse por produtos de origem vegetal. O objetivo deste artigo é revisar as substâncias isoladas de produtos naturais que apresentaram atividade moluscicida contra a espécie Biomphalaria glabrata a fim de reavaliar os protótipos mais promissores e atualizar o progresso das pesquisas para a obtenção de um novo moluscicida. Realizamos buscas em bases de dados científicas, como Scientific Electronic Library Online (SciELO), Google acadêmico, PUBMED, Web of Science e Literatura Latina-Americana e do Caribe em Ciências da saúde (LILACS). De 2000 a 2022, utilizando as palavras-chave "substâncias isoladas", "atividade moluscicida" e "Biomphalaria glabrata". No presente estudo, foi possível observar 19 moléculas moluscicidas promissoras com concentração letal abaixo de 20 µg/mL. Destes isolados promissores, apenas 5 isolados tiveram o CL90 calculado e dentro do valor recomendado pela OMS: ácido benzóico, 2',4',6'-triidroxidihidrocalcona, ácido divaricático, piplartina e 2-hidroxi-1,4-naftoquinona (Lapachol). Concluímos que além de poucos resultados na área, as pesquisas não seguem o padrão metodológico (tempo de exposição e unidades de medida, teste de toxicidade), assim como não seguem um padrão na exposição do resultado (CL), não seguindo, em suma, o recomendado pela OMS.

Epidemiological, clinical and laboratory aspects of Angiostrongylus cantonensis infection: an integrative review.

This integrative literature review study analyzes the findings of the last 5 years of the parasite Angiostrongylus cantonensis. It is known that this nematode is found in the pulmonary arteries of rats, where it remains as a definitive host. From mice, the cycle spreads to animals, such as snails, which in contact with humans can trigger the disease. In humans, the parasite causes several neurological, abdominal manifestations and mainly meningitis. Based on the review of studies, its epidemiology shows worldwide distribution, although there are endemic cases for this parasite, such as asian countries. Laboratory findings generally showed altered CSF with turbidity, increased protein and eosinophilia, which generated meningeal signs in the patient, moreover MRI exams showed multiple alterations. Rare findings of the nematode in the eyeball, lung and signs of peritoneal inflammation were reported, which requires further studies to understand the whole pathophysiology. Finally, conservative treatment based on anthelmintics and anti-inflammatories brought good responses, although there are reports of deaths, which demonstrates the importance in the prevention and therapy of this disease.

Natural products as a control measure of the Achatina fulica (Gastropoda: Achatinidae).

Achatina fulica is a terrestrial mollusk known as the giant African snail that is related to environmental, economic, urban, and public health problems. As control measures for this mollusk, cooking salt (NaCl) and calcium oxide (CaO) are used, and baits are composed of metaldehyde. However, these measures have environmental toxicity and impact the soil. In this way, natural products have been tested on this mollusk to discover and develop a substance to combat this urban and agricultural pest. This article aims to evaluate studies involving natural products to control the population of Achatina fulica. Articles and works published in books were included in the present work. A total of 1,103 works were found during the search. Of these, 14 works met the objective of these review and were included in this article. The tests do not possess methodological standardization, do not have a maximum concentration to be considered active, or a maximum exposure time. A lack of standardization in the methodology of tests on A. fulica was observed. The performance of tests on other life stages of the mollusk, as well as tests that analyze other parameters, are essential. Only one article analyzed presented phytochemical analysis. No ecotoxicity tests were reported either. Some extracts showed promising results, highlighting the aqueous extract of Capsicum frutescens. More studies investigating the molluscicidal activity of natural products on A. fulica are needed. It is very relevant that the new studies present a phytochemical analysis of the tested extracts, as well as ecotoxicity studies.

Epidemiological, clinical and laboratory aspects of Angiostrongylus cantonensis infection: an integrative review / Aspectos epidemiológicos, clínicos e laboratoriais de infecção por Angiostrongylus cantonensis: uma revisão integrativa

This integrative literature review study analyzes the findings of the last 5 years of the parasite Angiostrongylus cantonensis. It is known that this nematode is found in the pulmonary arteries of rats, where it remains as a definitive host. From mice, the cycle spreads to animals, such as snails, which in contact with humans can trigger the disease. In humans, the parasite causes several neurological, abdominal manifestations and mainly meningitis. Based on the review of studies, its epidemiology shows worldwide distribution, although there are endemic cases for this parasite, such as asian countries. Laboratory findings generally showed altered CSF with turbidity, increased protein and eosinophilia, which generated meningeal signs in the patient, moreover MRI exams showed multiple alterations. Rare findings of the nematode in the eyeball, lung and signs of peritoneal inflammation were reported, which requires further studies to understand the whole pathophysiology. Finally, conservative treatment based on anthelmintics and anti-inflammatories brought good responses, although there are reports of deaths, which demonstrates the importance in the prevention and therapy of this disease.

Este estudo de revisão integrativa da literatura analisa os achados dos últimos 5 anos do parasita Angiostrongylus cantonensis. Sabe-se que esse nematoide é encontrado nas artérias pulmonares de ratos, onde permanece como hospedeiro definitivo. Dos camundongos, o ciclo se espalha para os animais, como os caracóis, que em contato com humanos podem desencadear a doença. Em humanos, o parasita causa diversas manifestações neurológicas, abdominais e principalmente meningite. Com base na revisão de estudos, sua epidemiologia mostra distribuição mundial, embora existam casos endêmicos para esse parasito, como países asiáticos. Os achados laboratoriais geralmente mostraram LCR alterado com turbidez, aumento de proteínas e eosinofilia, o que gerou sinais meníngeos nos pacientes, além disso, os exames de ressonância magnética mostraram múltiplas alterações. Foram relatados achados raros do nematoide no globo ocular, pulmão e sinais de inflamação peritoneal, o que requer mais estudos para o entendimento de toda a fisiopatologia. Por fim, o tratamento conservador à base de anti-helmínticos e anti-inflamatórios trouxe boas respostas, embora haja relatos de óbitos, o que demonstra a importância na prevenção e terapia dessa doença.

Physalin pool from Physalis angulata L. leaves and physalin D inhibit P2X7 receptor function in vitro and acute lung injury in vivo.

P2X7 receptor promotes inflammatory response and neuropathic pain. New drugs capable of impairing inflammation and pain-reducing adverse effects extracted from plant extracts have been studied. Physalis angulate L. possesses traditional uses and exhibits antiparasitic, anti-inflammatory, antimicrobial, antinociceptive, antimalarial, antileishmanial, immunosuppressive, antiasthmatic. diuretic, and antitumor activities. The most representative phytochemical constituents identified with medicinal importance are the physalins and withanolides. However, the mechanism of anti-inflammatory action is scarce. Although some physalins and withanolides subtypes have anti-inflammatory activity, only four physalins subtypes (B, D, F, and G) have further studies. Therefore, we evaluated the crude ethanolic extract enriched with physalins B, D, F, and G from P. angulata leaves, a pool containing the physalins B, D, F, G, and the physalins individually, as P2X7 receptor antagonists. For this purpose, we evaluated ATP-induced dye uptake, macroscopic currents, and interleukin 1-ß (IL-1ß) in vitro. The crude extract and pool dose-dependently inhibited P2X7 receptor function. Thus, physalin B, D, F, and G individually evaluated for 5'-triphosphate (ATP)-induced dye uptake assay, whole-cell patch-clamp, and cytokine release showed distinct antagonist levels. Physalin D displayed higher potency and efficacy than physalin B, F, and G for all these parameters. In vivo mice model as ATP-induced paw edema was potently inhibited for physalin D, in contrast to physalin B, F, and G. ATP and lipopolysaccharide (LPS)-induced pleurisy in mice were reversed for physalin D treatment. Molecular modeling and computational simulation predicted the intermolecular interactions between the P2X7 receptor and physalin derivatives. In silico results indicated physalin D and F as a potent allosteric P2X7 receptor antagonist. These data confirm physalin D as a promisor source for developing a new P2X7 receptor antagonist with anti-inflammatory action.

Hypoxia enhances ILC3 responses through HIF-1α-dependent mechanism.

Group 3 innate lymphoid cells (ILC3) have a prominent role in the maintenance of intestine mucosa homeostasis. The hypoxia-inducible factor (HIF) is an important modulator of immune cell activation and a key mechanism for cellular adaptation to oxygen deprivation. However, its role on ILC3 is not well known. In this study, we investigated how a hypoxic environment modulates ILC3 response and the subsequent participation of HIF-1 signaling in this process. We found increased proliferation and activation of intestinal ILC3 at low oxygen levels, a response that was phenocopied when HIF-1α was chemically stabilized and was reversed when HIF-1 was blocked. The increased activation of ILC3 relied on a HIF-1α-dependent transcriptional program, but not on mTOR-signaling or a switch to glycolysis. HIF-1α deficiency in RORyt compartment resulted in impaired IL-17 and IL-22 production by ILC3 in vivo, which reflected in a lower expression of their target genes in the intestinal epithelium and an increased susceptibility to Clostridiodes difficile infection. Taken together, our results show that HIF-1α activation in intestinal ILC3 is relevant for their functions in steady state and infectious conditions.

Increased pregnancy rate in beef heifers resynchronized with estradiol at 14 days after TAI.

We aimed to evaluate the treatment with estradiol benzoate (EB) or 17ß-estradiol (E2) associated with progesterone (P4) for resynchronization of ovulation 14 days after timed artificial insemination (TAI). In Experiment 1 (Exp. 1), Nelore heifers were submitted to TAI (D0). On D14, the animals received an intravaginal P4 device and were randomly assigned to one of three groups: control (no treatment; n = 17); EB (1  mg EB; n = 17); and E2+P4 (1 mg E2 + 9 mg P4; n = 18). Ultrasonography evaluations were performed daily from D14 to D22 to map follicular and luteal dynamics. On D22, the P4 devices were removed and non-pregnant (NP) animals were determined using corpus luteum blood flow Doppler ultrasonography. In Exp. 2, 1295 beef heifers were resynchronized and randomly allocated to the same experimental groups as described in Exp. 1. On D22, the largest follicle (LF) was measured in NP and a second TAI was performed on D24. In a subset of heifers (n = 337), an estrus detection patch was used between D22 and D24 to monitor estrus expression and the LF was measured at D24. Confirmatory diagnosis of pregnancy was performed between D37-67 and D43-67 after first and second TAI, respectively. In Exp 1, the proportion of heifers with a synchronized follicular wave emergence (from 3 to 5 days after treatment) was greater (P < 0.05) in the EB group (93.8%) than in the control (62.5%) and E2+P4 (64.7%) groups. Structural luteolysis occurred earlier (P < 0.05) in the EB and E2+P4 groups than in the controls. The pregnancy rate after first TAI did not differ (P > 0.1) among the groups at D22 and at confirmatory diagnosis in both experiments. In Exp 2, the potential pregnancy loss between D22 and D37-67 was similar (P > 0.1) in the control (19% [36/185]), EB (15% [28/182]) and E2+P4 (15% [28/184]) groups. The LF diameter (mm) on D22 was greater (P < 0.05) in the control group (11.9 ± 0.1) than in EB (11.3 ± 0.1) and E2+P4 (11.5 ± 0.1). No difference (P > 0.1) was observed in the proportion of heifers detected in estrus, but LF growth rate (mm/day) between D22 and D24 was greater (P < 0.05) in EB group (0.9 ± 0.08) than in control (0.6 ± 0.07) and E2+P4 (0.7 ± 0.09) groups. The pregnancy rate for the second TAI was greater (P < 0.05) in the EB group (47% [94/200]) than in the control (37% [76/203]), but did not differ (P > 0.1) from the E2+P4 group (43% [93/214]). In conclusion, the treatment with 1 mg EB or 1 mg E2 + 9 mg P4 at 14 days post-TAI anticipates luteolysis in NP heifers but does not compromise pregnancy. The EB treatment induces a new synchronized follicle wave emergence and increases the pregnancy rate of resynchronized NP heifers.

Terrestrial and marine Antarctic fungi extracts active against Xanthomonas citri subsp. citri.

This study aims to obtain secondary metabolites extracts from filamentous fungi isolated from soil and marine sediments from Antarctica and assess its potential antibacterial activity on Xanthomonas citri subsp. citri, the agent of citrus canker. Metabolites production was conducted in Malt 2% broth at 15°C for 20 days after which intracellular and extracellular extracts were obtained. The extracts were evaluated by cell viability assays through Resazurin Microtitre Assay. From 158 fungal extracts, 33 hampered bacterial growth in vitro. The average inhibition of the extracts obtained from terrestrial (soil) and marine (sediments) fungi was 94 and 97% respectively. These inhibition values were close to the average of 90% cell death for the positive control. MIC90 and MBC for the bioactive extracts were established. Isolates that produced active metabolites against the phytopathogen were identified using molecular taxonomy (ITS-rRNA sequencing) as: Pseudogymnoascus, Penicillium, Cadophora, Paraconiothyrium and Toxicocladosporium. Antarctic fungal strains isolated from terrestrial and marine sediments were able to produce secondary metabolites with antimicrobial activity against X. citri subsp. citri, highlighting the importance of these microbial genetic resources. These metabolites have potential to be used as alternatives for the control of this plant pathogen. SIGNIFICANCE AND IMPACT OF THE STUDY: This manuscript makes an impact on the study of micro-organisms from extreme habitats and their possible contribution in discovering new active molecules against pathogens of agricultural interest. Studies on the Antarctic continent and its communities have attracted the scientific community due to the long period of isolation and low levels of disturbance that surrounds the region. Knowing the potential of fungi in this region to produce active secondary metabolites, we aim to contribute to the discovery of compounds with antibacterial action in Xanthomonas citri subsp. citri, a plant pathogen present in several regions around the globe.

Performance, carcass and meat characteristics of two cattle categories finished on pasture during the dry season with supplementation in different forage allowance / Desempenho, características da carcaça e carne de duas categorias de bovinos de corte terminados em pastagem durante a estação seca com suplementação em diferentes ofertas de forragem

The objective this paper was to evaluate the effect of two categories of beef finished in pasture with supplementation with two herbage allowance on performance, carcass and meat characteristics. Thirty-six Guzera cattle were used, 18 steers and 18 heifers with an initial age of 20 months. There was significant difference in daily weight gain for animal category and the herbage allowance, which were higher in males and animals submitted to high herbage allowance. Steers showed higher final weight, carcass weight and forequarter yield compared with the heifers, although the hindquarter yield was higher in the heifers, however the herbage allowance did not influence these characteristics. There were not statistical differences for carcass yield, ribeye area, backfat thickness and marbling score for the animal categories and herbage allowance. The meat chemical composition of the steers did not differ of the heifers, however, the animals submitted to high herbage allowance was increase in ether extract and pH, decrease in protein percentage. It was concluded that the animal category and the herbage allowance changed the animal performance, improving performance in males and cattle submitted to high herbage allowance.(AU)

O objetivo do presente trabalho foi avaliar o efeito de duas categorias de bovinos terminados em pastagem com suplementação em duas ofertas de forragem no desempenho, na qualidade da carcaça e da carne. Foram utilizados 36 bovinos Guzerá, 18 novilhos e 18 novilhas, com idade inicial de 20 meses de idade. Observou-se diferença significativa no ganho de peso diário para categoria animal e ofertas de forragem, que foram superiores nos machos e nos animais submetidos à oferta alta de forragem. Os novilhos apresentaram maior peso final, peso de carcaça e rendimento de dianteiro em comparação com as novilhas, embora o rendimento do traseiro tenha sido maior para as novilhas, entretanto as ofertas de forragem não influenciaram essas características. Não houve diferenças estatísticas para rendimento de carcaça, área de olho de lombo, espessura de gordura subcutânea e marmorização para categorias de animais e ofertas de forragem. Quanto à composição química, a carne de novilhos não diferiu das novilhas. Os animais submetidos à alta oferta de forragem tiveram aumento no extrato etéreo e pH, redução na porcentagem de proteína. Conclui-se que a categoria animal e os níveis de forragem alteram o desempenho animal, melhorando, assim, o desempenho nos machos e nos bovinos submetidos à alta oferta de forragem.(AU)

Performance, carcass and meat characteristics of two cattle categories finished on pasture during the dry season with supplementation in different forage allowance / Desempenho, características da carcaça e carne de duas categorias de bovinos de corte terminados em pastagem durante a estação seca com suplementação em diferentes ofertas de forragem

The objective this paper was to evaluate the effect of two categories of beef finished in pasture with supplementation with two herbage allowance on performance, carcass and meat characteristics. Thirty-six Guzera cattle were used, 18 steers and 18 heifers with an initial age of 20 months. There was significant difference in daily weight gain for animal category and the herbage allowance, which were higher in males and animals submitted to high herbage allowance. Steers showed higher final weight, carcass weight and forequarter yield compared with the heifers, although the hindquarter yield was higher in the heifers, however the herbage allowance did not influence these characteristics. There were not statistical differences for carcass yield, ribeye area, backfat thickness and marbling score for the animal categories and herbage allowance. The meat chemical composition of the steers did not differ of the heifers, however, the animals submitted to high herbage allowance was increase in ether extract and pH, decrease in protein percentage. It was concluded that the animal category and the herbage allowance changed the animal performance, improving performance in males and cattle submitted to high herbage allowance.(AU)

O objetivo do presente trabalho foi avaliar o efeito de duas categorias de bovinos terminados em pastagem com suplementação em duas ofertas de forragem no desempenho, na qualidade da carcaça e da carne. Foram utilizados 36 bovinos Guzerá, 18 novilhos e 18 novilhas, com idade inicial de 20 meses de idade. Observou-se diferença significativa no ganho de peso diário para categoria animal e ofertas de forragem, que foram superiores nos machos e nos animais submetidos à oferta alta de forragem. Os novilhos apresentaram maior peso final, peso de carcaça e rendimento de dianteiro em comparação com as novilhas, embora o rendimento do traseiro tenha sido maior para as novilhas, entretanto as ofertas de forragem não influenciaram essas características. Não houve diferenças estatísticas para rendimento de carcaça, área de olho de lombo, espessura de gordura subcutânea e marmorização para categorias de animais e ofertas de forragem. Quanto à composição química, a carne de novilhos não diferiu das novilhas. Os animais submetidos à alta oferta de forragem tiveram aumento no extrato etéreo e pH, redução na porcentagem de proteína. Conclui-se que a categoria animal e os níveis de forragem alteram o desempenho animal, melhorando, assim, o desempenho nos machos e nos bovinos submetidos à alta oferta de forragem.(AU)

Frequency of Aedes sp. Linnaeus (Diptera: Culicidae) and Associated Entomofauna in Bromeliads from a Forest Patch within a densely Urbanized Area.

Little is known about the relationship between the presence of Aedes, abiotic factors and the entomofauna existing in phytotelmatas. The aim of this study was to identify biotic and abiotic factors associated with the presence of Aedes in bromeliads sites located in a forest fragment in Rio de Janeiro, Brazil. In the study area, eight bromeliads from the Aechmea genus were chosen and kept in landscape form. Physical and chemical variables were measured. Collected aquatic insects were identified according to the literature. A total of 3102 immature insects were collected. The presence of Aedes aegypti (Linnaeus) (2.29%) was rare. Few specimens were found concentrated in urban-adjacent areas during summer. On the other hand, Aedes albopictus (Skuse) (17.57%) was found throughout the year in 0%-80% of the sites, averaging 1.0 mosq/bromelia. Aedes albopictus was found predominantly in central sites of the forest fragment. The Canonical Correspondence Analysis indicates that most taxa had a moderate association with temperature, dissolved oxygen, and pH. The abiotic variables, such as temperature and dissolved oxygen, affect the distribution of the genus Aedes vectors, while most of the other variables did not.

DNase concentration assay to obtain DNA-free RNA from sugarcane leaves.

The success of gene expression studies, protein synthesis, and construction of cDNA libraries directly depends on the purity and integrity of the RNA used in these tests, as even minor amounts of contaminant DNA (<1%) can produce a false positive amplification signal in quantitative real-time PCR. For RNA contaminated with genomic DNA, an essential step in the studies on gene expression is the treatment of the RNA samples with DNase. This study was conducted to test three different concentrations of DNase I (0.02, 0.04, and 0.06 µL/​​ng of RNA), which were chosen based on the results of the RNA sample quantifications and as indicated by the manufacturer, to digest genomic DNA present in the RNA samples extracted from sugarcane leaves with the Concert™ Plant RNA Reagent. The results showed that all three concentrations of DNase significantly reduced DNA concentrations. However, RNA was also degraded on DNase I treatment. In addition, the amount of DNA present in the RNA samples after purification with DNase I was sufficient for its amplification in the tests conducted with conventional PCR. Furthermore, the condition of RNA samples obtained after the treatments allowed for real-time PCR. Therefore, we concluded that 0.02 µL DNase I was the ideal concentration for sugarcane RNA purification, as higher concentrations do not increase the efficiency of the genomic DNA digestion in RNA samples and only make the purification process more expensive. This study provides important information on the effect of high concentrations of DNase I and complements previous studies that have so far tested only the DNase concentration recommended by the manufacturer.

Genetic aspects of Huntington's disease in Latin America. A systematic review.

We aimed to present a systematic review on Huntington's disease (HD) in Latin America (LA). PubMed and LILACS were searched up to March 2015, reporting confirmed HD cases in LA. Case series, cross-sectional, case-control, and prospective studies were included. From 534 communications, 47 were eligible. Population-based studies were not found; minimal prevalence of 0.5-4/100,000 was estimated for Venezuela and Mexico. Geographical isolates were well characterized in Venezuela and in Peru. CAG repeats at HTT gene varied between 7-33 and 37-112 in normal and expanded alleles, respectively. Intermediate alleles were found in 4-10% of controls. Ages at onset and the expanded CAG repeats correlated with r from - 0.55 to -0.91. While haplotype patterns of Venezuelan and Brazilian chromosomes were similar to those observed in Europeans, haplotypes from Peruvian HD patients did not match the same pattern. The limited number of papers found suggests that HD is poorly diagnosed in LA. Minimal prevalence seemed to be halfway between those of Caucasians and Asians. Range of CAG repeats was similar to those of Europeans. Haplotype studies indicate that majority of HD patients might be of Caucasian descent; an Asian origin for some Peruvian patients was proposed.

MTHFR C677T and A1298C polymorphisms as predictors of radiotherapy response in head and neck squamous cell carcinoma.

The C677T and A1298C polymorphisms in methylene-tetrahydrofolate reductase (MTHFR), which regulates the release of active folate in the body, may have reduced activity. Given that folate participates in important intracellular pathways, such as nucleotide synthesis and biomolecule methylation, it seems plausible that patients with head and neck squamous cell carcinoma (HNSCC) may respond differently to radiotherapy treatments, based on genetic polymor-phisms. Therefore, this study sought to understand the role of these polymorphisms in HNSCC patient radiotherapy response. Genotypes were detected by PCR-RFLP after extraction of DNA from peripheral blood lymphocytes. Survival curves were analyzed by the Kaplan- Meier model, and significant differences were analyzed by the Wil-coxon test. Response to radiotherapy in patients with laryngeal SCC was significantly associated with the MTHFR C677T polymorphism (P = 0.030). Indeed, the presence of at least one T allele decreases the mortality rate up to 3-fold. Therefore, we propose that MTHFR C677T may represent a putative biomarker for radiotherapy prognosis in la-ryngeal SCC patients.

Prognostic significance of head and neck squamous cell carcinoma repair gene polymorphism.

The aims of this study were to analyze the polymorphisms XRCC1 Arg194Trp, XRCC1 Arg399Gln, XRCC3 Thr241Met, XPC Lys939Gln, ERCC1 Asn118Asn, and RAD51 -98G>C and to verify their influence on radiotherapy response and prognosis of patients with head and neck squamous cell carcinoma (HNSCC). Peripheral blood DNA was extracted from 311 patients and analyzed by PCR-RFLP. Our results showed that in irradiated oral and oropharyngeal patients, the 939Gln allele increased 6-fold local disease relapse risk (OR = 6.04; CI = 1.47-24.88) and over 2-fold the earliness of relapse (HR = 2.63; CI = 1.04-6.70). As for the XRCC3 polymorphism, multivariate analysis showed that the 241Met allele increases over 33-fold local relapse risk (OR = 33.64; CI = 3.23-350.85), over 12-fold earliness of relapse (HR = 12.55; CI = 2.47-63.73) and over 3-fold earliness of death (HR = 3.04; CI = 1.08-8.61). For polymorphism RAD51 -98, multivariate analysis showed that allele C increases over 3-fold the risk of relapse (OR = 3.13; CI = 1.12-8.78) and over 2-fold the earliness of relapse (HR = 2.84; CI = 1.25-6.47). For polymorphism XRCC1 Arg399Gln, multivariate analysis showed that the 399Gln allele increased the risk of local disease relapse for irradiated oral and oropharyngeal patients (OR = 3.35; CI = 1.10-10.13) by over 3-fold. Based on these results, we suggest that these polymorphisms may be useful markers of prognosis in HNSCC.

Avaliação histomorfométrica do efeito do extrato aquoso de urucum (norbixina) no processo de cicatrização de feridas cutâneas em ratos / Histomorphometric evaluation of the effects of the aqueous extract of annatto (norbixin) on wound skin healing in rats

A norbixina é um carotenóide dicarboxílico hidrofílico presente no pericarpo das sementes do urucuzeiro (Bixa orellana L.). O urucum é comumente utilizado na indústria alimentícia e cosmética como corante natural. O objetivo deste trabalho foi avaliar o processo de reparo cicatricial de feridas cutâneas abertas de ratos tratadas com extrato aquoso de urucum contendo 2,5% de norbixina através de análise histomorfométrica. Para tanto, realizou-se feridas cutâneas no dorso de 32 ratos Wistar machos. Estas foram tratadas com extrato de urucum (grupo experimental, n = 16) e solução salina 0,9% (grupo controle, n = 16). Aos 3, 7, 14 e 21 dias após realização do procedimento cirúrgico, os animais foram sacrificados. Os preparados histológicos obtidos foram submetidos à técnica de coloração pela Hematoxilina-Eosina (H.E.) para contagem de células inflamatórias e de fibroblastos; corados pelo azul de toluidina 1% para contagem dos mastócitos e com picrossirius-red para avaliação das fibras colágenas totais. As imagens histológicas destas lâminas foram capturadas por câmera digital acoplada ao microscópio óptico, sob foco fixo e clareza de campo, obtendo-se 10 campos por lâmina com aumento final de 400X. As fotomicrografias foram avaliadas através do software ImageJ. Os resultados obtidos foram submetidos ao teste t de student sendo o valor de p considerado significativo para p< 0,05. O tratamento tópico com extrato de urucum utilizado aumentou o infiltrado inflamatório durante o 3º, 14º e 21º dia pós-operatório. O grupo tratado apresentou maior densidade vascular quando comparado ao controle a partir do 7º e menor quantidade de fibroblastos até o 14º dia pós-cirúrgico. Conclui-se que o extrato de urucum contendo 2,5% de norbixina não é inócuo aos tecidos cutâneos e possui efeitos pró-inflamatórios e pró-angiogênicos durante o processo de reparo tecidual cutâneo em ratos, interferindo no processo fisiológico de cicatrização.

Norbixin is a dicarboxylic water-soluble carotenoid present in the pericarp of the achiote (Bixa orellana L.) seed. Annatto is commonly used by the food and cosmetic industries as a natural pigment. The aim of this study was to evaluate the healing process of open dermal wounds of rats treated with an aqueous solution of annatto containing 2.5% of norbixin carried out by histomorphometric analysis. For this end, cutaneous wounds were made on the back of 32 male Wistar rats. The wounds were treated with annatto solution (experimental group, n = 16) and saline solution 0.9% (control group, n = 16). At 3, 7, 14 and 21 days after the surgical procedure, the animals were sacrificed. After histological preparation, the histological material was submitted to the staining technique with hematoxylin-eosin for the counting of inflammatory cells and fibroblasts, with toluidine blue 1% for mast cell counting and with picrosirius red for the total collagen fibers. The images of those histological slides were captured by a digital camera connected to an optical microscope, with fixed focal length and clear field; 10 fields were captured from each slide with final zoom of 400X. The photomicrographs were analyzed by the ImageJ software. The results were submitted to the Student's t-test and the value of p < 0.05 was considered as relevant. The topical treatment with the annatto solution increased the inflammatory infiltrate during the 3rd, 14th and 21st days after surgery. The experimental group presented increased vascular density compared to the control group after the 7th day and a smaller amount of fibroblasts up to the 14th day after surgery. We concluded that the annatto solution containing 2.5% of norbixin is not innocuous to skin tissues and has proinflammatory and proangiogenic effects during the process of skin wound healing in rats, interfering in the physiological healing process.

Research opportunities in intraoperative radiation therapy: the next decade 2013-2023.

The reality of intraoperative radiation therapy (IORT) practice is consistent with an efficient and highly precise radiation therapy technique to safely boost areas at risk for local recurrence. Long-term clinical experience has shown that IORT-containing multi-modality regimens appear to improve local disease control, if not survival in many diseases. Research with IORT is a multidisciplinary scenario that covers knowledge from radiation beam adapted development to advance molecular biology for bio-predictability of outcome. The technical parameters employed in IORT procedures are important information to be recorded for quality assurance and clinical results analysis. In addition, specific treatment planning systems for IORT procedures are available, to help in the treatment decision-making process. A systematic revision of opportunities for research and innovation in IORT is reported including radiation beam modulation, delivery, dosimetry and planning; infrastructure and treatment factors; experimental and clinical radiobiology; clinical trials, innovation and translational research development.