A pcr-based method for SARS-COV-2 variant detection in wastewater.

McAdam, Ewan - Associate SupervisorThe COVID-19 outbreak, caused by the SARS-CoV-2 virus, rapidly evolved into a worldwide pandemic, as declared by World Health Organization on 11th March 2020. The continued spread since early 2020 has resulted in many variants of this virus. Most mutations found along its genome are known as single nucleotide polymorphisms (SNPs) where only one base pair is mutated. Current real-time qualitative polymerase chain reaction (RT-qPCR) protocols, so called gold-standard method, are used to confirm if a person is positive or negative for the virus. There is a lack of available technology for rapid identification of variants. In order to identify the presence of a viral variant it is necessary to perform sequencing. Sequencing is expensive and may take hours to days to complete. Due to its cost, sequencing is widely unavailable in most countries. Even in countries where sequencing is available, like the UK, the number of samples sequenced are less than 10% of total cases due to extremely high cost. Wastewater-based epidemiology (WBE) is a novel approach that would help monitor and possibly revert the current health crisis. It has been reported the presence of SARS-CoV-2 RNA in faeces of infected individuals. This makes it possible to detect and monitor SARS-CoV-2 in wastewater samples, providing a health status report of the population within the catchment. To this context, WBE also enables to monitor the dissemination of variants for early warning of the outbreak within the defined population. In this project, a RT-qPCR method was developed targeting unique SNPs of SARS-CoV-2. This assay uses two probes, both targeting the same sequence, one with the SNP and the other non- SNP. These mutations are found in the N-gene of SARS-CoV-2 viral RNA, a conserved region that contains unique SNPs specific to each variant for differentiation. By using two probes, binding competition occurs, and the differentiation is done by observing an earlier detection with the SNP probe (~6 cycles). In addition to this, this method can be coupled with a melt curve analysis for further confirmation. Currently, there is a lack of available technology for rapid identification of variants of concern within the community. This assay can be implemented for routine WBE. By developing a SNP-PCR assay to detect specific variants of concern using WBE, it would be possible to accurately detect variants. This information provides a comprehensive health report on the population that could possibly help revert the current health crisis.MSc by Research in Wate

Os Refugiados no Brasil e as Organizações não Governamentais

In this century the flow of refugees and immigrants will become the central issue to International Human Rights Law. Brazil, that recently started to receive a larger flow of refugees, is an important option for them due to its benevolent legislation. However, the Brazilian State is still reluctant concerning the impact of the newly arrived refugees that enter the country without receiving subsequent support. This deductive research, in the historical type, intends to analyze the pioneer role of human rights NGOs responsible for informing and supporting the refugees, offering visibility to the theme.Neste século o fluxo de refugiados e imigrantes tornar-se-á a questão central para o Direito Internacional dos Direitos Humanos. O Brasil, que começou recentemente a receber um fluxo maior de refugiados, é uma opção importante para eles em razão de sua legislação benevolente. No entanto, o Estado brasileiro ainda é relutante quanto ao impacto das recentes levas de refugiados que entram no país sem receber o devido apoio posteriormente. Esta pesquisa dedutiva, na modalidade histórica, pretende analisar o papel pioneiro das ONGs de direitos humanos responsáveis por informar e apoiar os refugiados, oferecendo visibilidade ao tema

Protetor solar: um aliado na prevenção de efeitos causados pelos raios nocivos do sol

Os protetores solares são substâncias que protegem a pele contra a ação dos raios ultravioleta do Sol. Essas substâncias com proteção não eficazes provocam vermelhidão da pele. O objetivo principal deste trabalho é investigar se os professores de diferentes áreas de conhecimento do CA/UFSC conhecem as novas normas criadas pela Agência Nacional de Vigilância Sanitária (ANVISA) para a fabricação de protetores solares. Nesse sentido, os dados foram coletados através de questionário contendo questões de sondagem acerca da utilização de protetor solar, do horário de exposição ao Sol, do número do FPS utilizado e da preocupação com os efeitos de uma exposição ao Sol sem proteção; uma notícia para leitura: “Agência Nacional de Vigilância Sanitária cria normas para fabricação de protetor solar”, divulgada em 04 de junho de 2012 no Telejornal Jornal Nacional da Rede Globo de Televisão na forma impressa e questões relacionadas à notícia. Os dados foram analisados através de gráficos. Posteriormente, com base nestes resultados, foi elaborado um guia prático de cuidados antes, durante e após a exposição ao Sol. Palavras-chave: Protetor solar; Sol; Saúde

Ética e moral na escola: o que os documentos regulares dizem a respeito?

Objetiva-se identificar quais são os documentos legais que orientam ética e moralmente as práticas pedagógicas dos educadores escolares e quais seriam essas orientações dentro da concepção de educação vigente em nosso país. Conceitua-se o que é moral e o que é ética em nossa sociedade, com base na obra de Piaget. Compreendemos que um ambiente moral na escola, pode favorecer o convívio coletivo, pois a moral autônoma pode ser desenvolvida com mais facilidade. Escolhemos os seguintes documentos para estudo: Diretrizes Curriculares do Curso de Pedagogia (2006), dos Parâmetros Curriculares Nacionais (1997), e do Plano Nacional de Educação (2014). Após a análise dos documentos fizemos uma breve discussão a respeito do papel do professor e da educação moral dos alunos. Por fim, pudemos concluir com base nos documentos estudados e no referencial teórico escolhido, que quando o ambiente educacional e profissional se organiza a partir da adoção da ética e da moral nos processos pedagógicos, os resultados tendem a ser positivos, pois, além de valorizar e desenvolver a competência do professor, a ênfase se faz sobre relações cooperativas, que podem promover o desenvolvimento moral autônomo do aluno

Antimicrobial Peptides Designed against the Ω-Loop of Class A β-Lactamases to Potentiate the Efficacy of β-Lactam Antibiotics

Class A serine β-lactamases (SBLs) have a conserved non-active site structural domain called the omega loop (Ω-loop), in which a glutamic acid residue is believed to be directly involved in the hydrolysis of β-lactam antibiotics by providing a water molecule during catalysis. We aimed to design and characterise potential pentapeptides to mask the function of the Ω-loop of β-lactamases and reduce their efficacy, along with potentiating the β-lactam antibiotics and eventually decreasing β-lactam resistance. Considering the Ω-loop sequence as a template, a group of pentapeptide models were designed, validated through docking, and synthesised using solid-phase peptide synthesis (SPPS). To check whether the β-lactamases (BLAs) were inhibited, we expressed specific BLAs (TEM-1 and SHV-14) and evaluated the trans-expression through a broth dilution method and an agar dilution method (HT-SPOTi). To further support our claim, we conducted a kinetic analysis of BLAs with the peptides and employed molecular dynamics (MD) simulations of peptides. The individual presence of six histidine-based peptides (TSHLH, ETHIH, ESRLH, ESHIH, ESRIH, and TYHLH) reduced β-lactam resistance in the strains harbouring BLAs. Subsequently, we found that the combinational effect of these peptides and β-lactams sensitised the bacteria towards the β-lactam drugs. We hypothesize that the antimicrobial peptides obtained might be considered among the novel inhibitors that can be used specifically against the Ω-loop of the β-lactamases

Regulatory T Cells Accumulate in the Lung Allergic Inflammation and Efficiently Suppress T-Cell Proliferation but Not Th2 Cytokine Production

Foxp3+CD25+CD4+ regulatory T cells are vital for peripheral tolerance and control of tissue inflammation. In this study, we characterized the phenotype and monitored the migration and activity of regulatory T cells present in the airways of allergic or tolerant mice after allergen challenge. To induce lung allergic inflammation, mice were sensitized twice with ovalbumin/aluminum hydroxide gel and challenged twice with intranasal ovalbumin. Tolerance was induced by oral administration of ovalbumin for 5 consecutive days prior to OVA sensitization and challenge. We detected regulatory T cells (Foxp3+CD25+CD4+ T cells) in the airways of allergic and tolerant mice; however, the number of regulatory T cells was more than 40-fold higher in allergic mice than in tolerant mice. Lung regulatory T cells expressed an effector/memory phenotype (CCR4highCD62LlowCD44highCD54highCD69+) that distinguished them from naive regulatory T cells (CCR4intCD62LhighCD44intCD54intCD69−). These regulatory T cells efficiently suppressed pulmonary T-cell proliferation but not Th2 cytokine production

Regulatory T Cells Accumulate in the Lung Allergic Inflammation and Efficiently Suppress T-Cell Proliferation but Not Th2 Cytokine Production

Foxp3+CD25+CD4+ regulatory T cells are vital for peripheral tolerance and control of tissue inflammation. In this study, we characterized the phenotype and monitored the migration and activity of regulatory T cells present in the airways of allergic or tolerant mice after allergen challenge. To induce lung allergic inflammation, mice were sensitized twice with ovalbumin/aluminum hydroxide gel and challenged twice with intranasal ovalbumin. Tolerance was induced by oral administration of ovalbumin for 5 consecutive days prior to OVA sensitization and challenge. We detected regulatory T cells (Foxp3+CD25+CD4+ T cells) in the airways of allergic and tolerant mice; however, the number of regulatory T cells was more than 40-fold higher in allergic mice than in tolerant mice. Lung regulatory T cells expressed an effector/memory phenotype (CCR4highCD62LlowCD44highCD54highCD69+) that distinguished them from naive regulatory T cells (CCR4intCD62LhighCD44intCD54intCD69−). These regulatory T cells efficiently suppressed pulmonary T-cell proliferation but not Th2 cytokine production

Recognition and behavioral assessment of acute pain in cats: literature review

Embora a dor seja considerada o quarto sinal vital e uma das manifestações mais comumente encontradas na prática médica veterinária dos animais domésticos, seu tratamento ainda é inadequado. A dor aguda pós-operatória tem suscitado grande interesse por seu potencial risco de cronificação caso não adequadamente tratada, podendo piorar a recuperação e a qualidade de vida do paciente. O gato é uma das espécies domésticas menos estudadas no que diz respeito ao reconhecimento e controle da dor, e algumas das dificuldades residem na avaliação e na percepção da dor. O consenso sobre os sinais comportamentais da dor nesta espécie publicado em fevereiro de 2016 considerou alguns sinais como confiáveis e sensíveis para a avaliação da dor em gatos, em toda uma gama de diferentes condições clínicas, porém afirma a necessidade da realização de estudos que analisem a sua validade e aplicabilidade clínica, especialmente em relação a diferentes intensidades de dor. Na tentativa de se quantificar a dor são utilizados vários tipos de escalas subjetivas tradicionais e outras que facilitam a avaliação da efetividade da analgesia, a partir da observação de sinais comportamentais espontâneos indicativos de dor, combinada a uma resposta qualitativa à palpação da ferida cirúrgica. Faz-se necessária a utilização de escalas específicas para o tipo de dor (aguda ou crônica) e para a espécie, de modo a minimizar a subjetividade e a parcialidade dos observadores e possibilitando uma melhor assistência ao paciente.Although pain is considered the 4th vital sign and one of the most frequently observed clinical signs in domestic animals’ clinical practice, its treatment is still inadequate despite significant improvement in the last few years. Acute post-operative pain has aroused great interest due to its potential risk of developing into chronic pain, and if not treated properly, it might worsen the recovery and the patient’s quality of life. Cats are one of the least studied species of domestic animals regarding pain recognition and control. Some of the difficulties lie in pain assessment and perception. The consensus published in February 2016 about behavioral signs of pain in cats considered some signs to be reliable and sensitive for the assessment of pain in this species in many different clinical conditions, however it still states that more studies will be necessary in order to evaluate its clinical validity and applicability, especially considering the various pain intensities. As an attempt to quantify pain intensity in cats, several types of traditional subjective scales and others that facilitate pain assessment by combining the observation of spontaneous behavioral signals of pain and qualitative response to palpation of surgical wound are used as tools. It is necessary to use specific scales for each type of pain and for each specific animal species so to minimize the subjectivity and the partiality of the observers, reducing bias and improving efficacy, thus leading to a better patient care

Estudo anatômico e histoquímico das partes vegetativas de Sansevieria trifasciata Prain

Sansevieria trifasciata Prain, pertence à família Asparagaceae Juss, conhecida popularmente como espada-de-São-Jorge, lança-de-São-Jorge, espadinha, língua-de-sogra, sanseveria, é uma herbácea, nativa da África, utilizada como ornamental e na medicina popular. O estudo objetivou caracterizar a anatomia do rizoma, folha e escapo floral e caracterizar a histoquímica da folha, visando aprofundar o conhecimento dessa espécie. Foram confeccionadas lâminas semipermanentes com secções transversais do rizoma, folha e escapo floral, além de secções longitudinais do rizoma e paradérmicas da lâmina foliar, que foram analisadas em microscopia óptica de luz e microscopia de polarização. Para estudo histoquímico foram utilizadas lâminas semipermanentes com secções transversais da folha. As caracterizações anatômicas evidenciaram a identificação de caracteres importantes, tais como ráfides e canais secretores no rizoma. A presença de alcaloides, compostos fenólicos, compostos lipofílicos e lignina foi observada em testes histoquímicos. Os resultados contribuem para o conhecimento farmacobotânico, no controle da qualidade da espécie estudada, evitando o uso inadequado e adulteração, no uso de espécies de S. trifasciata, com o nome vulgar espada-de-São-Jorge

Estudo da Compressão Primária de Resíduos Sólidos Urbanos (RSU) Aterrados / Study of the Primary Compression of Grounded Municipal Solid Waste (MSW)

The monitoring of differential settlements is used to estimate the increased lifecycle of sanitary landfills, to maintain slope stabilization and safety of operations. The research objective was to use the soil mechanics methodology for determining the physical indexes used to calculate the settlement of Municipal Solid Waste (MSW). MSW samples were collected in a landfill in Brazil, at two different depths - Group 1 (0.5 m), Group 2 (0.8 m). Samples had ten years of grounding. Group 1 presented an average moisture content of 121.43%; specific mass of 1.06 ± 0.24 g.cm-³; initial void ratio of 5.73 ± 1.65; final void ratio of 4.10 ± 1.42; and primary compression index (Cc) of 1.26 ± 0.26. Group 2 presented an average moisture content value of 118.66%; specific mass of 1.10 ± 0.4 g.cm-³; initial void ratio of 6.72 ± 3.9; final void ratio of 5.13 ± 3.1; and Cc of 1.32 ± 0.26. No significant differences were identified between the Cc’s of Group 1 and 2, although the different depths. The methodology in this study can be validated, as it shows that soil mechanics characteristics can be applied to MSW, since the results were similar to other studies in the literature.