Spatial and temporal variation of fungal endophytic richness and diversity associated to the phyllosphere of olive cultivars

Fungal endophytes are micro-organisms that colonize healthy plant tissues without causing disease symptoms. They are described as plant growth and disease resistance promoters and have shown antimicrobial activity. The spatial-temporal distribution of endophytic communities in olive cultivars has been poorly explored. This study aims to investigate the richness and diversity of endophytic fungi in different seasons and sites, within the Alentejo region, Portugal. Additionally, and because the impact of some pathogenic fungi (e.g. Colletotrichum spp.) varies according to olive cultivars; three cultivars, Galega vulgar, Cobrançosa and Azeiteira, were sampled. 1868 fungal isolates were identified as belonging to 26 OTUs; 13 OTUs were identified to the genera level and 13 to species level. Cultivar Galega vulgar and season autumn showed significant higher values in terms of endophytic richness and diversity. At site level, Elvas showed the lowest fungal richness and diversity of fungal endophytes. This study reinforces the importance of exploring the combined spatio-temporal distribution of the endophytic biodiversity in different olive cultivars. Knowledge about endophytic communities may help to better understand their functions in plants hosts, such as their ecological dynamics with pathogenic fungi, which can be explored for their use as biocontrol agents

Development and Validation Dissolution Analytical Method of Nimesulide beta-Cyclodextrin 400 mg Tablet

The nimesulide (N-(4-nitro-2-phenoxyphenyl)methanesulfonamide) belongs to the class of non-steroidal anti-inflammatory drugs (NSAIDs and category II of the biopharmaceutical classification, The complexation of nimesulide with b-cyclodextrin is a pharmacological strategy to increase the solubility of the drug The objective of this study was to develop and validate an analytical methodology for dissolving the nimesulide beta-cyclodextrin 400 mg tablet and meets the guidelines of ANVISA for drug registration purposes. Once developed, the dissolution methodology was validated according to the RE of parameters no.  899/2003. In the development of the method it was noted that the duration of the dissolution test was 60 minutes, the volume and the most suitable dissolution medium was 900 mL of aqueous solution of sodium lauryl sulfate 1% (w/ v). It was also noted that rotation of 100 rpm and the paddle apparatus was the most appropriate to evaluate the dissolution of the drug. Spectrophotometric methodology was used to quantify the percentage of dissolved drug. The wavelength was 390 nm using the quantification. The validation of the methodology, system suitability parameters, specificity/selectivity, linearity, precision, accuracy and robustness were satisfactory and proved that the developed dissolution methodology was duly executed. DOI: http://dx.doi.org/10.17807/orbital.v8i5.82

PROTOTHECA ZOPFII: IMPORTANTE PATÓGENO NA ETIOLOGIA DA MASTITE BOVINA NO BRASIL

A alga Prototheca zopfii destaca-se entre os agentes ambientais da mastite bovina em virtude dos prejuízos decorrentes do comprometimento da qualidade e produção de leite. Além disso, apresenta elevada resistência antimicrobiana e potencial zoonótico, fatores que levam ao descarte dos animais infectados. Esse patógeno foi identificado recentemente em Goiás e acredita-se que possa estar presente em outros rebanhos do Estado. O presente estudo visa revisar a literatura referente às características do agente, aspectos econômicos e de saúde pública, epidemiologia, clínicos e patológicos, bem como os procedimentos para diagnóstico, controle e prevenção dessa forma de mastite. PALAVRAS-CHAVE: Controle de mastite, epidemiologia, Prototheca zopfii

Effect of long-term fungicide applications on virulence and diversity of Colletotrichum spp. Associated to olive anthracnose

In this study, the presence and variability of Colletotrichum spp. was evaluated by comparing fungal isolates obtained from olive trees under long-time phytosanitary treatments with trees without any phytosanitary treatments (treated and untreated, respectively). Olive fruits of trees of the highly susceptible ‘Galega vulgar’ cultivar growing in the Alentejo region were used as samples. From the 210 olive trees sampled (half from treated and half from untreated orchards), 125 (59.5%) presented Colletotrichum spp., with a significant lower number of infected trees in treated (39) when compared to untreated orchards (86). The alignment and analysis of beta-tubulin (tub2), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), actin (ACT), chitin synthase (CHS-1) and histone H3 (HIS-3) gene sequences allowed the identification of all 125 isolates as belonging to the C. acutatum complex. The vast majority of the isolates (124) were identified as C. nymphaeae and one isolate, from an untreated tree, was identified as C. godetiae. Isolates were divided into five different groups: Group A: 39 isolates from treated trees matched in 100% with C. nymphaeae sequences from the database; Group B: 76 isolates from untreated trees matched in 100% with C. nymphaeae sequences from the database; Group C: one isolate from untreated trees presenting a single nucleotidic difference in the HIS-3 sequence; Group D: eight isolates from untreated trees presenting differences in two nucleotides in the tub2 sequences that changed the protein structure, together with differences in two specific nucleotides of the GAPDH sequences; Group E: one isolate, from untreated olive trees, matched 100% with C. godetiae sequences from the database in all genes. Considering the similarities of the sampled areas, our results show that the long-time application of fungicides may have caused a reduction in the number of olive trees infected with Colletotrichum spp. but an increase in the number of fruits positive to Colletotrichum spp. within each tree, which may suggest different degrees of virulence of Colletotrichum isolates from trees growing different management regimes. It is imperative that the fungicides described as causing resistance are applied at appropriate times and intervals, since their efficiency decreases when applied incorrectly and new and more virulent species may arise

A Metabolomics Study

Funding Information: This work is financed by national funds from Fundação para a Ciência e a Tecnologia (FCT), I.P., in the scope of the project UIDP/04378/2020 and UIDB/04378/2020 of the Research Unit on Applied Molecular Biosciences (UCIBIO) and the project LA/P/0140/2020 of the Associate Laboratory Institute for Health and Bioeconomy—i4HB and through the project EXPL/MEDFAR/0203/2021. A. Dias-Carvalho acknowledges FCT and UCIBIO for her PhD grant (UI/BD/151318/2021). V.M.C acknowledges FCT for her grant (SFRH/BPD/110001/2015) that was funded by national funds through FCT under the Norma Transitória–DL57/2016/CP1334/CT0006. A.R.-M. acknowledges FCT for her grant SFRH/BD/129359/2017. Publisher Copyright: © 2023 by the authors.Long-term cognitive dysfunction, or “chemobrain”, has been observed in cancer patients treated with chemotherapy. Mitoxantrone (MTX) is a topoisomerase II inhibitor that binds and intercalates with DNA, being used in the treatment of several cancers and multiple sclerosis. Although MTX can induce chemobrain, its neurotoxic mechanisms are poorly studied. This work aimed to identify the adverse outcome pathways (AOPs) activated in the brain upon the use of a clinically relevant cumulative dose of MTX. Three-month-old male CD-1 mice were given a biweekly intraperitoneal administration of MTX over the course of three weeks until reaching a total cumulative dose of 6 mg/kg. Controls were given sterile saline in the same schedule. Two weeks after the last administration, the mice were euthanized and their brains removed. The left brain hemisphere was used for targeted profiling of the metabolism of glutathione and the right hemisphere for an untargeted metabolomics approach. The obtained results revealed that MTX treatment reduced the availability of cysteine (Cys), cysteinylglycine (CysGly), and reduced glutathione (GSH) suggesting that MTX disrupts glutathione metabolism. The untargeted approach revealed metabolic circuits of phosphatidylethanolamine, catecholamines, unsaturated fatty acids biosynthesis, and glycerolipids as relevant players in AOPs of MTX in our in vivo model. As far as we know, our study was the first to perform such a broad profiling study on pathways that could put patients given MTX at risk of cognitive deficits.publishersversionpublishe

Progressive Histiocytosis of Non-Epitheliotropic Dendritic Cells in a Feline

Background: Histiocytic tumors in felines are nodules that commonly develop on limbs and head extremities. They can be divided into many subtypes including cutaneous histiocytoma, histiocytic sarcoma, reactive fibrohistiocytic nodule, Langerhans cell histiocytosis, and progressive feline dendritic cell. Despite the same origin, they have behaviors that differ from each other, thus it is important to confirm diagnosis with histopathological and immunohistochemical tests, because early identification can facilitate prognosis and treatment. In this study, we describe the pathological and immunohistochemical characteristics, enabling differentiation feline neoplasms derived from histiocytes. Case: A 5-year-old, crossbreed, male, feline presented with a nodulation at the base of the left ear. The mass was slow growing, partially alopecic, with no other changes associated with tumor development. The nodule was round and circumscribed, movable, with an elevated surface. He was referred for surgery and an elliptical sample around the tumor was carefully dissected. Routine histopathological evaluation was performed with hematoxylin and eosin (HE), as well as immunohistochemistry. Histopathology showed circumscribed proliferation of histiocytic cells, with abundant and eosinophilic cytoplasm. The proliferative cells were large and rounded, extending from the superficial dermis and basement membrane to the deep dermis. At the extremities, some cells had visible vacuoles. Mitotic activity ranged from 3 to 4 mitoses per field in 40x magnification. Immunohistochemistry showed positive staining for histocompatibility complex MCII and lysozyme antibodies, marking histiocytic cells. Labeling was positive for CD20 in cells of lymphoid lineage B and negative for E-cadherin. Histiocytic cells did not invade the epidermis; hence, proliferation was classified as nonepitheliotropic. These methods contribute to the literature regarding the diagnosis of this rare tumor. Therefore, histological as well as immunohistochemical evaluation are important bfor confirming clinical diagnosis of histiocytic proliferation non-epitheliotropic. Discussion: Progressive histiocytosis of feline dendritic cells, in both epitheliotropic and non-epitheliotropic forms, is considered a clinically progressive and rare disorder. There are reports which include cytological, clinical, histological and immunohistochemical examinations, but the diagnostic characteristics regarding the non-epitheliotropic classification have not yet been properly identified. Nodulations are predominantly observed in head and limb regions, usually non-ulcerated, which can both increase and decrease in size, and are typically painless. The tumor in the present case was restricted to the base of the ear and no evidence of infiltration or metastasis was found. Progressive histiocytosis may spread and reach the lymphatic system through the lymph nodes, subsequently becoming systemic. The non-aggressive behavior observed in this case is possibly related to the non-epitheliotropic pattern. In the present case, MHC II histocompatibility complex markers, a phenotype compatible with dendritic cells, were used. Lysozyme antibodies marked histiocytic cells and the reactive lymphoid infiltrate was composed of CD20-positive B lymphoid lineage cells. Staining for E-cadherin was negative, negative results in labeling experiments is common, it is dependent upon the cellular origin of the leukocytes present in the sample. Staining for these molecules is recommended for differentiating feline progressive histiocytosis from Langerhans cells. Langerhans cells can be characterized by E-cadherin expression in about 10% of cases and marked T lymphocyte and neutrophil expression in the affected tissue. In this case, the histopathological exam along with immunohistochemistry was essential for differentiating them

Development and Validation Dissolution Analytical Method of Nimesulide beta-Cyclodextrin 400 mg Tablet

The nimesulide (N-(4-nitro-2-phenoxyphenyl)methanesulfonamide) belongs to the class of non-steroidal anti-inflammatory drugs (NSAIDs and category II of the biopharmaceutical classification, The complexation of nimesulide with b-cyclodextrin is a pharmacological strategy to increase the solubility of the drug The objective of this study was to develop and validate an analytical methodology for dissolving the nimesulide beta-cyclodextrin 400 mg tablet and meets the guidelines of ANVISA for drug registration purposes. Once developed, the dissolution methodology was validated according to the RE of parameters no.  899/2003. In the development of the method it was noted that the duration of the dissolution test was 60 minutes, the volume and the most suitable dissolution medium was 900 mL of aqueous solution of sodium lauryl sulfate 1% (w/ v). It was also noted that rotation of 100 rpm and the paddle apparatus was the most appropriate to evaluate the dissolution of the drug. Spectrophotometric methodology was used to quantify the percentage of dissolved drug. The wavelength was 390 nm using the quantification. The validation of the methodology, system suitability parameters, specificity/selectivity, linearity, precision, accuracy and robustness were satisfactory and proved that the developed dissolution methodology was duly executed. DOI: http://dx.doi.org/10.17807/orbital.v8i5.82

Infusions of Portuguese medicinal plants: dependence of final antioxidant capacity and phenol content on extraction features

BACKGROUND: Aqueous extracts of most medicinal plants traditionally employed in Portugal (at the ratio of 1 g plant: 110mL water) have been assayed for total antioxidant capacity and phenol content, in order to elucidate their claimed medicinal features. RESULTS: The antioxidant activity was assessed by theABTS•+ method; the ascorbic acid equivalent values ranged from 1.4280 ± 0.1261 g L−1 for avocado (Persea americana (Lauraceae)) obtained by infusion of powder, down to 0.0027 ± 0.0012 g L−1 for olive (Olea europaea (Oleaceae)) obtained by infusion of leaves. Total phenol content was determined by the Folin–Ciocalteu procedure; the gallic acid equivalent values ranged from 0.5541 ± 0.0289 g L−1 for avocado obtained by infusion of powder, down to 0.0053 ± 0.0014 g L−1 for olive obtained by boiling leaves. A good correlation between total antioxidant capacity and total phenol content was found. CONCLUSION: The method of powder infusion should be chosen if high concentration of antioxidants are sought. On the other hand, a high antioxidant capacity and a high phenol content correlate well with the empirically established (and widely publicised) capacity to treat respiratory infections

Knowledge, attitude and practices of biomedicine students on the papanicolaou exam

Cervical cancer is the second type of cancer that most affects women, considered a serious public health problem, however it can be detected and treated early through the Pap smear. This research aimed to analyze the knowledge, attitude and practices of biomedicine students on the Pap smear. A cross-sectional study on Knowledge, Attitude and Practice (KAP), applied with 108 students, was carried out at a Higher Education Institution in Teresina, Piauí, Brazil. The data were collected through the months of February and March of 2018 with the application of a questionnaire. Statistical analysis was descriptive in terms of absolute and relative frequencies. Knowledge 72 (66.6%) and attitude 76 (70.3%) presented adequate levels regarding the examination. As for the practice, the percentage of suitability was of 37 academics (34.2%) relatively not adequate. It was concluded that the students had adequate knowledge and attitude, but an inadequate practice, and the Papanicolaou test is required for the early screening and prevention of cervical cancer

Núcleo de atendimento on-line dos indivíduos com microcefalia durante a pandemia pela COVID-19: relato de experiência

Em decorrência da pandemia pela COVID-19, é indiscutível a existência de inúmeros desafios para a saúde pública. O propósito deste estudo foi relatar experiências dos discentes da Universidade Estadual do Sudoeste da Bahia participantes do projeto extensionista de promoção da saúde oral aos indivíduos com microcefalia no formato remoto. Trata-se da elaboração de folhetos educativos e vídeos sobre saúde oral e desenvolvimento dentário pelos 6 graduandos do curso de Odontologia e enviados pelo aplicativo WhatsApp aos cuidadores dos indivíduos com microcefalia de julho a dezembro de 2020. Vinte e seis famílias participaram com interação do conteúdo elaborado e a construção de uma comunicação digital com estimulação favorável dos indivíduos com microcefalia dos hábitos de higiene oral, acompanhamento do desenvolvimento dentário e musicoterapia. Esta vivência resultou em ganho científico, ético e humano dos alunos envolvidos. A extensão universitária mais do que nunca deve assumir sua responsabilidade social, construindo novos caminhos para execução dos projetos a fim de minimizar os impactos da pandemia na vida da sociedade