Introduction, Dispersal, and Predominance of SARS-CoV-2 Delta Variant in Rio Grande do Sul, Brazil: A Retrospective Analysis.

Mutations in the SARS-CoV-2 genome can alter the virus' fitness, leading to the emergence of variants of concern (VOC). In Brazil, the Gamma variant dominated the pandemic in the first half of 2021, and from June onwards, the first cases of Delta infection were documented. Here, we investigate the introduction and dispersal of the Delta variant in the RS state by sequencing 1077 SARS-CoV-2-positive samples from June to October 2021. Of these samples, 34.7% were identified as Gamma and 65.3% as Delta. Notably, 99.2% of Delta sequences were clustered within the 21J lineage, forming a significant Brazilian clade. The estimated clock rate was 5.97 × 10-4 substitutions per site per year. The Delta variant was first reported on 17 June in the Vinhedos Basalto microregion and rapidly spread, accounting for over 70% of cases within nine weeks. Despite this, the number of cases and deaths remained stable, possibly due to vaccination, prior infections, and the continued mandatory mask use. In conclusion, our study provides insights into the Delta variant circulating in the RS state, highlighting the importance of genomic surveillance for monitoring viral evolution, even when the impact of new variants may be less severe in a given region.

Pipeline validation for the identification of antimicrobial-resistant genes in carbapenem-resistant Klebsiella pneumoniae.

Antimicrobial-resistant Klebsiella pneumoniae is a global threat to healthcare and an important cause of nosocomial infections. Antimicrobial resistance causes prolonged treatment periods, high mortality rates, and economic impacts. Whole Genome Sequencing (WGS) has been used in laboratory diagnosis, but there is limited evidence about pipeline validation to parse generated data. Thus, the present study aimed to validate a bioinformatics pipeline for the identification of antimicrobial resistance genes from carbapenem-resistant K. pneumoniae WGS. Sequences were obtained from a publicly available database, trimmed, de novo assembled, mapped to the K. pneumoniae reference genome, and annotated. Contigs were submitted to different tools for bacterial (Kraken2 and SpeciesFinder) and antimicrobial resistance gene identification (ResFinder and ABRicate). We analyzed 201 K. pneumoniae genomes. In the bacterial identification by Kraken2, all samples were correctly identified, and in SpeciesFinder, 92.54% were correctly identified as K. pneumoniae, 6.96% erroneously as Pseudomonas aeruginosa, and 0.5% erroneously as Citrobacter freundii. ResFinder found a greater number of antimicrobial resistance genes than ABRicate; however, many were identified more than once in the same sample. All tools presented 100% repeatability and reproducibility and > 75% performance in other metrics. Kraken2 was more assertive in recognizing bacterial species, and SpeciesFinder may need improvements.

Development and application of a mini-QuEChERS method for the determination of pesticide residues in anuran adipose tissues.

The expansion of monocultures to regions close to conservation areas has put biodiversity at risk, mainly due to the intense use of pesticides. Anurans are highly susceptible to pesticides and may be a biological marker in the contamination of an area. However, methods for determining pesticides in anurans are incipient. In this work, a miniaturized QuEChERS method was developed for the extraction of atrazine, chlorpyrifos, α- and ß-endosulfan, α-, ß-, θ- and ζ-cypermethrin in anuran adipose tissues. The method was optimized for the tissue sample size scale according to sample mass availability. Extracting solvent and adsorbents for the clean-up step was evaluated, achieving recoveries next to 100% with acetonitrile and without a clean-up step. The mini-QuEChERS method, using 500 mg of adipose tissue, 50 mg of NaCl and 200 mg of MgSO4, 100 µL of ultrapure water, and 1.50 mL of acetonitrile with no purification step, followed by high-performance liquid chromatography analysis and photodiode array detection was validated following the European Community guidelines. The methodology showed a moderate matrix effect for some pesticides, which was corrected using the matrix-matched calibration. The limits of quantification for the pesticide residues in adipose tissues ranged from 10 to 75 µg kg-1. Pesticide recoveries ranged from 74% to 115%, and repeatability and within-lab reproducibility showed relative standard deviations < 11%. The mini-QuEChERS method was applied to extract pesticide residues from the adipose tissues of two species of anurans: Leptodactylus macrosternum and Scinax x-signatus. 25% of samples were positive, detecting endosulfan and chlorpyriphos, confirmed by liquid chromatography coupled to tandem mass spectrometry. The mini-QuEChERS was a simple, economical, and eco-friendly method for extracting pesticide residues in anuran adipose tissue samples.

Incipient Parallel Evolution of SARS-CoV-2 Deltacron Variant in South Brazil.

With the coexistence of multiple lineages and increased international travel, recombination and gene flow are likely to become increasingly important in the adaptive evolution of SARS-CoV-2. These processes could result in genetic introgression and the incipient parallel evolution of multiple recombinant lineages. However, identifying recombinant lineages is challenging, and the true extent of recombinant evolution in SARS-CoV-2 may be underestimated. This study describes the first SARS-CoV-2 Deltacron recombinant case identified in Brazil. We demonstrate that the recombination breakpoint is at the beginning of the Spike gene. The 5' genome portion (circa 22 kb) resembles the AY.101 (Delta), and the 3' genome portion (circa 8 kb nucleotides) is most similar to the BA.1.1 (Omicron). Furthermore, evolutionary genomic analyses indicate that the new strain emerged after a single recombination event between lineages of diverse geographical locations in December 2021 in South Brazil. This Deltacron, AYBA-RS, is one of the dozens of recombinants described in 2022. The submission of only four sequences in the GISAID database suggests that this lineage had a minor epidemiological impact. However, the recent emergence of this and other Deltacron recombinant lineages (XD, XF, and XS) suggests that gene flow and recombination may play an increasingly important role in the COVID-19 pandemic. We explain the evolutionary and population genetic theory that supports this assertion, concluding that this stresses the need for continued genomic surveillance. This monitoring is vital for countries where multiple variants are present, as well as for countries that receive significant inbound international travel.

How the speech-language pathologist interacts with cystic fibrosis patients? a scoping review / Qual a relação da fonoaudiologia com a fibrose cística? uma revisão de escopo / ¿Cuál es la relación entre la logopedia y la fibrosis quís- tica? una revisión del alcance

Objective: The review describes the changes related to sleep-breathing disorders, com- munication and feeding processes in subjects with CF, through a scoping review. Meth- ods: A scoping review based on the PRISMA protocol was carried out in Pubmed, Lilacs and Scopus databases, to investigate the possibility of including SLP in the treatment of patients with CF. The inclusion criteria included studies that addressed communication and eating processes and disorders related to CF. Studies with a sample that presented other comorbidities that would justify the worsening of the condition, or secondary stud- ies, were excluded. There was no time or language delimitation. Results: The search found 1566 works, 27 Lilacs, 1009 Pubmed and 530 Scopus, by analysis of titles, abstract and reading in full. A total of 30 articles were selected for inclusion, 2 Lilacs, 22 Pubmed and 6 Scopus, all related to Speech-Language Pathology. Conclusion: It was observed that there is a poor sleep quality due to nocturnal desaturation, mild and moderate obstructive apnea. In studies related to hearing, individuals had sensorineural hearing loss. Speech-Language Pathology is of paramount importance in monitoring these sub- jects.

Objetivo: Descrever alterações relacionadas aos processos de distúrbios respiratórios do sono, comunicação e alimentação em sujeitos com FC, por meio de uma scoping review. Métodos: Foi realizada uma scoping review nas bases de dados Pubmed, Lilacs e Scopus, para investigar a possibilidade de incluir o fonoaudiólogo no tratamento de pacientes com FC, utilizando o checklist PRISMA. Os critérios de inclusão incluíram estudos que abor- dassem processos de comunicação e alimentação e distúrbios relacionados à FC. Foram excluídos estudos com amostra que apresentasse outras comorbidades que justificassem o agravamento do quadro, ou estudos secundários. Não houve delimitação de tempo ou idioma. Resultados: A busca encontrou 1566 trabalhos, sendo 27 Lilacs, 1009 Pubmed e 530 Scopus, por meio de análise de títulos, resumo e leitura na íntegra. Foram seleciona- dos para inclusão 30 artigos, sendo 2 Lilacs, 22 Pubmed e 6 Scopus, todos relacionados à Fonoaudiologia. Conclusão: Observou-se que há má qualidade do sono devido à dessa- turação noturna, apneia obstrutiva leve e moderada. Em estudos relacionados à audição, os indivíduos apresentaram perda auditiva neurossensorial. A Fonoaudiologia é de suma importância no acompanhamento desses sujeitos.

Objetivo: Describir las alteraciones relacionadas con los procesos de trastornos respira- torios del sueño, comunicación y alimentación en sujetos con FQ, a través de una revisión de alcance. Métodos: Se realizó una revisión de alcance en las bases de datos Pubmed, Lilacs y Scopus, para investigar la posibilidad de incluir al logopeda en el tratamiento de pacientes con FQ, utilizando la lista de verificación PRISMA. Los criterios de inclusión incluyeron estudios que abordaran los procesos de comunicación y alimentación y los trastornos relacionados con la FQ. Se excluyeron los estudios con una muestra que pre- sentaba otras comorbilidades que justificasen el empeoramiento del cuadro, o estudios secundarios. No había delimitación de tiempo ni de idioma. Resultados: La búsqueda en- contró 1566 trabajos, de los cuales 27 fueron Lilacs, 1009 Pubmed y 530 Scopus, me- diante análisis de títulos, resúmenes y lectura completa. Se seleccionaron 30 artículos para inclusión, 2 Lilacs, 22 Pubmed y 6 Scopus, todos relacionados con Logopedia. Con- clusión: Se observó que existe mala calidad del sueño por desaturación nocturna, apnea obstructiva leve y moderada. En estudios relacionados con la audición, los individuos tenían pérdida auditiva neurosensorial. La logopedia es de suma importancia en el segui- miento de estos sujetos.

Cross comparison of seminal plasma proteins from cattle and buffalo (Bubalus bubalis).

The objective of this study was to evaluate seminal plasma proteins from cattle and buffalo (Bubalus bubalis), to identify differences between related species. Sixteen buffaloes and 16 cattle between 30 and 60 months of age were used. Semen collection was performed by electroejaculation, followed by macroscopic and microscopic subjective analyses. After analysis, the samples were centrifuged at 800 g for 10 min, and the supernatant (seminal plasma) was recentrifuged at 10,000 g for 30 min at 4°C. The total protein concentration was determined by the Bradford method, and the proteins were digested in solution for mass spectrometry (nLC-MS/MS). Multivariate statistical analysis was used to evaluate the proteomics results by non-hierarchical clustering the considering exponentially modified protein abundance index (emPAI). Principal component analysis (PCA) and partial least squares discriminant analysis (PLS-DA) were used for clustering. Proteomics identified 78 proteins, and multivariate analysis showed 4 that were over-expressed in buffaloes (cystatin C, prosaposin, peptide YY and keratin type II cytoskeletal 5) and 9 in cattle (spermadhesin-1, seminal plasma protein PDC-109, ribonuclease 4, metalloproteinase inhibitor 2, acrosin inhibitor 1, seminal ribonuclease, C-type natriuretic peptide, angiogenin-1 and osteopontin). Among the proteins identified in seminal plasma, the C-type natriuretic peptide and metalloproteinase inhibitors were described for the first time in buffaloes. Some protease inhibitors were found over-expressed in buffaloes, and important proteins in seminal plasma of cattle were not identified or were found at lower expression levels in buffaloes, which can contribute to reproductive performance in this species.

Proteomic analysis of amniotic and allantoic fluid from buffaloes during foetal development.

The objective of this study was to describe the dynamic changes in protein composition and protein abundance in amniotic and allantoic fluids from buffaloes during gestation. Amniotic and allantoic fluids were collected during the first, second and third trimesters of gestation. The foetuses were measured and weighed. Fluid samples were centrifuged at 800 g for 10 min and then at 10,000 g for 60 min at 4°C. The supernatant was collected to determine the total protein concentration. Based on total protein concentration, an aliquot (50 µg) was used for in-solution tryptic digestion, and mass spectrometry analysis (nano-LC-MS/MS) was performed. A multivariate statistical analysis of the proteomic data was conducted. Across the different stages of buffalo gestation, fifty-one proteins were found in the amniotic fluid, and twenty-one were found in the allantoic fluid. A total of twelve proteins were common among the stages, and four presented significant differences (VIP score α > 1). Fibronectin and alpha-1-antiproteinase were more abundant in the amniotic fluid than in the allantoic fluid. Alpha-2-macroglobulin and alpha-2-HS-glycoprotein were more abundant in the allantoic fluid than in the amniotic fluid. Alpha-2-macroglobulin participates in remodelling and growth of the uterus at beginning of the gestation (first trimester), and these findings indicate that can serve as a potential tool for the early diagnosis of pregnancy in buffaloes.

Endometriose umbilical: relato de caso e revisão de literatura / Umbilical endometriosis: a case report and literature review

A endometriose é caracterizada pela presença de tecido endometrial fora da cavidade uterina. Seu mecanismo fisiopatológico é pouco esclarecido, mas várias hipóteses são descritas - dentre elas a disseminação de células endometriais por via hematogênica ou linfática. A apresentação clínica é bastante variável, mas podem estar presentes massa palpável, sangramentos cíclicos ou alterações de coloração da pele na região afetada. O diagnóstico é estabelecido a partir do exame clínico, associado a exames de imagem e biópsia da lesão. O tratamento de escolha é o cirúrgico com excisão da lesão e reconstrução da anatomia local. Apresentamos um relato de caso de uma paciente de 35 anos, com endometriose umbilical, com necessidade de excisão da lesão e confecção de novo umbigo. A evolução da afecção descrita é estritamente relacionada ao manejo diagnóstico e propedêutico instituído; sendo assim, a exclusão de diagnósticos diferenciais, benignos ou malignos, é de suma importância.

Endometriosis is characterized by the presence of endometrial tissue outside the uterine cavity. The pathophysiology of this condition is poorly understood. However, several hypotheses have been proposed, including the spread of endometrial cells via hematogenous or lymphatic routes. The clinical presentation is variable but includes a palpable nodule, cyclic bleeding, and changes in the color of the skin in the affected region. Diagnosis is established by clinical examination combined with imaging and lesion biopsy. The treatment of choice is surgical, with excision of the lesion and tissue reconstruction. We report the case of a 35-year-old female patient with umbilical endometriosis and the need for removal of the lesion with umbilical reconstruction. The presentation of this condition determines the diagnostic and management approach. Therefore, the exclusion of other benign or malignant disorders is essential.

Stationary phase based on cellulose dodecanoate physically immobilized on silica particles for high-performance liquid chromatography.

The chemical agent free preparation of a stationary phase using a natural macromolecule was the focus of this paper. Thermal immobilization of cellulose dodecanoate on silica particles was used for the preparation of a stationary phase without the use of chemical reagents. Cellulose modification was performed to produce a hydrophobic macromolecule with solubility in common organic solvents. The new stationary phase was characterized morphologically and physico-chemically, presenting as spherical particles immobilized with a thin cellulose dodecanoate layer. The degree of substitution of cellulose dodecanoate was 1.7, which resulted in a separation mechanism in reversed phase mode, but with lower hydrophobicity and higher steric selectivity, which are properties from cellulose. These characteristics resulted in a stationary phase with intrinsic selectivity that was able to separate mixtures of polar drugs, homologs of an anionic surfactant and omeprazole isomers, which are not well resolved in typical C18 phases. Considering that cellulose is a natural polymer and the preparation method of stationary phase involves only physical processes of silica modification, the final material presents as a stationary phase with specific retention properties coming from both dodecanoate and cellulose.

Preparation and Characterization of Cellulose Triacetate as Support for Lecitase Ultra Immobilization.

The use of polymers as supports for enzyme immobilization is a strategy that enables to remove the enzymes from a chemical reaction and improve their efficiency in catalytic processes. In this work, cellulose triacetate (CTA) was used for physical adsorption of phospholipase Lecitase ultra (LU). CTA is more hydrophobic than cellulose, shows good performance in the lipases immobilization being a good candidate for immobilization of phospholipases. We investigated the immobilization of LU in CTA, the stability of the immobilized enzyme (CTA-LU) and the performance of CTA-LU using soybean oil as a substrate. LU was efficiently immobilized in CTA reaching 97.1% in 60 min of contact with an enzymatic activity of 975.8 U·g-1. The CTA-LU system presents good thermal stability, being superior of the free enzyme and increase of the catalytic activity in the whole range of pH values. The difference observed for immobilized enzyme compared to free one occurs because of the interaction between the enzyme and the polymer, which stabilizes the enzyme. The CTA-LU system was used in the transesterification of soybean oil with methanol, with the production of fatty acid methyl esters. The results showed that CTA-LU is a promising system for enzymatic reactions.

A simple and fast method for the production and characterization of methylic and ethylic biodiesels from tucum oil via an alkaline route.

A simple, fast, and complete route for the production of methylic and ethylic biodiesel from tucum oil is described. Aliquots of the oil obtained directly from pressed tucum (pulp and almonds) were treated with potassium methoxide or ethoxide at 40°C for 40 min. The biodiesel form was removed from the reactor and washed with 0.1 M HCl aqueous solution. A simple distillation at 100°C was carried out in order to remove water and alcohol species from the biodiesel. The oxidative stability index was obtained for the tucum oil as well as the methylic and ethylic biodiesel at 6.13, 2.90, and 2.80 h, for storage times higher than 8 days. Quality control of the original oil and of the methylic and ethylic biodiesels, such as the amount of glycerin produced during the transesterification process, was accomplished by the TLC, GC-MS, and FT-IR techniques. The results obtained in this study indicate a potential biofuel production by simple treatment of tucum, an important Amazonian fruit.

Análise de voz e comunicação oral de professores de curso pré-vestibular / Voice and oral communication analysis of preparatory school teachers

OBJETIVO: Realizar uma auto-avaliação com professores de curso pré-vestibular enfocando aspectos de voz, bem como analisar as características de voz, fala e recursos corporais utilizados em sala de aula. MÉTODOS: Participaram deste estudo 40 professores do sexo masculino de uma empresa de cursos pré-vestibulares. Os docentes foram avaliados no local de trabalho por meio de dois questionários. O primeiro referente à auto-avaliação vocal e do ambiente de trabalho e o segundo referente à avaliação perceptivo-auditiva e visual de parâmetros corporais. A avaliação perceptivo-auditiva e visual foi realizada pela pesquisadora durante aulas-padrão de 60 minutos. Os parâmetros analisados foram divididos em aspectos de voz e fala. Os parâmetros referentes à análise dos recursos corporais foram pontuados em uma escala analógica graduada de 0 a 10, sendo que nesta escala o extremo esquerdo (0) refere-se à inadequação e o extremo direito (10) à completa adequação. RESULTADOS: A maioria dos professores relatou usar microfone (77,5 por cento) e apresentaram queixas de sinais e sintomas vocais, sendo os mais citados: falhas na voz, rouquidão, esforço para falar, secura na garganta e pigarro. Na análise perceptivo-auditiva observou-se que os professores apresentaram qualidade vocal discretamente alterada, sendo 35 por cento do tipo rouca. Os aspectos de fala e recursos corporais foram utilizados na maior parte do tempo de forma adequada. CONCLUSÃO: Os professores de curso pré-vestibular apresentaram elevada incidência de sintomas vocais, mesmo com a utilização de microfone. Utilizaram diversos recursos corporais na tentativa de potencializar o aprendizado, sendo que o uso de gestos, contato visual direcionado e interação verbal com os alunos foram as estratégias mais utilizadas em sala de aula.

PURPOSE: To carry out a self-evaluation with preparatory school teachers regarding vocal aspects, as well as to analyze characteristics of voice, speech and corporal resources used during classes. METHODS: Forty male teachers from a preparatory school were selected for this study. They were assessed in their workplace using two questionnaires. The first questionnaire regarded the workplace and vocal self-evaluation, and the second concerned visual and auditory-perceptual assessment of corporal parameters. The visual and auditory-perceptual assessment was carried out by the examiner during regular 60-minute classes. The parameters analyzed were divided into voice and speech aspects. The parameters that regarded corporal resources analysis were scored on an analogical scale graduated from 0 to 10, where the far left (0) referred to inadequacy, and the far right (10), to complete adequacy. RESULTS: Most teachers (77.5 percent) used microphone during classes and reported vocal signs and symptoms, among which the most cited were: voice failures, hoarseness, speech effort, throat dryness and accumulation of mucus in the throat. Auditory-perceptual analysis showed that teachers had slightly altered vocal quality, being 35 percent characterized as hoarse. Speech and corporal resources were used adequately most of the time. CONCLUSION: Prep-school teachers showed high incidence of vocal symptoms even when using microphone. They used of several corporal resources in order to enhance learning, and use of gestures, directed eye contact, and verbal interaction with students were the most used strategies during classes.