Structural and functional analysis of four non-coding Y RNAs from Chinese hamster cells: identification, molecular dynamics simulations and DNA replication initiation assays.

BACKGROUND: The genes coding for Y RNAs are evolutionarily conserved in vertebrates. These non-coding RNAs are essential for the initiation of chromosomal DNA replication in vertebrate cells. However thus far, no information is available about Y RNAs in Chinese hamster cells, which have already been used to detect replication origins and alternative DNA structures around these sites. Here, we report the gene sequences and predicted structural characteristics of the Chinese hamster Y RNAs, and analyze their ability to support the initiation of chromosomal DNA replication in vitro. RESULTS: We identified DNA sequences in the Chinese hamster genome of four Y RNAs (chY1, chY3, chY4 and chY5) with upstream promoter sequences, which are homologous to the four main types of vertebrate Y RNAs. The chY1, chY3 and chY5 genes were highly conserved with their vertebrate counterparts, whilst the chY4 gene showed a relatively high degree of diversification from the other vertebrate Y4 genes. Molecular dynamics simulations suggest that chY4 RNA is structurally stable despite its evolutionarily divergent predicted stem structure. Of the four Y RNA genes present in the hamster genome, we found that only the chY1 and chY3 RNA were strongly expressed in the Chinese hamster GMA32 cell line, while expression of the chY4 and chY5 RNA genes was five orders of magnitude lower, suggesting that they may in fact not be expressed. We synthesized all four chY RNAs and showed that any of these four could support the initiation of DNA replication in an established human cell-free system. CONCLUSIONS: These data therefore establish that non-coding chY RNAs are stable structures and can substitute for human Y RNAs in a reconstituted cell-free DNA replication initiation system. The pattern of Y RNA expression and functionality is consistent with Y RNAs of other rodents, including mouse and rat

Development and implantation of PCR-SSP for the genotyping of JAK2 V617F mutation / Desenvolvimento e implantação de metodologia molecular baseada em PCR-SSP para genotipagem da mutação V617F de JAK2

The JAK2 protein promotes cells growth and proliferation, and mutations in the JAK2 gene can result in increase of the number of blood cells and in development of myeloproliferative neoplasms. This study aimed to develop and implement the PCR-SSP (Sequence-Specific Primer - Polymerase Chain Reaction) to detect the JAK2V617F mutation. Therefore, a literature review about genotyping methodologies for this mutation was conducted and primers were based on the model proposed by Xavier (2009), tested and adjusted to the best amplification condition. The PCR-SSP technique was standardized and was effective for the detection of the JAK2V617F mutation. As it is not expensive, the technique reduces costs and can be implanted even in small molecular biology laboratories, helping the diagnosis of patients with myeloproliferative diseases and favoring research related to this gene

TELEATENDIMENTO EM PLATAFORMA DIGITAL COMO FERRAMENTA EDUCACIONAL: PERCEPÇÕES DE ACADÊMICOS DA ÁREA DA SAÚDE

Devido a pandemia do Coronavírus, países estão adotando medidas preventivas quanto a sua disseminação, sendo uma destas medidas o distanciamento social para evitar exposição desnecessária a ambientes que podem ter o vírus circulante. Neste sentido, uma das alternativas que está sendo desenvolvida é a telemedicina. O estado do Paraná desenvolveu uma plataforma digital para teleconsulta e telemonitoramento, sendo os atendimentos realizados por bolsistas e profissionais. Considerando que o atendimento remoto é uma tecnologia a pouco inserida no contexto do cuidado e da educação, este presente relato tem por objetivo apresentar a plataforma digital como ferramenta educacional para os bolsistas e para a população atendida, sob a ótica dos acadêmicos participantes. De acordo com as experiências vivenciadas, a plataforma contribui para a educação em saúde da população ao se realizar orientações de medidas preventivas, bem como contribui para a formação acadêmica dos participantes, ao estimular o desenvolvimento de conhecimentos, habilidades e atitudes para realizar atendimento adequado. Assim, conclui-se que a telemedicina contribui para a promoção de saúde entre a população e para a formação de profissionais de saúde mais críticos-reflexivos

Plagiarism as another ethical issue in scientific research

The excessive demand for publications results in high plagiarism and duplicate numbers by scientists who take over existing texts into new publications. In addition to serious ethical problems, this practice hinders the generation of original material. In order to reduce the problem, softwares such as eTBLAST are being used to detect plagiarism and repeated papers. Despite the persistence of fraudsters, these tools have helped to reduce these problems; however, the ideal solution would be the basic ethical establishment principles. Therefore, plagiarism has always been a foible that could lead to fraudulent and dishonorable development of science

Sequencing, cloning, and heterologous expression of cyclomaltodextrin glucanotransferase of Bacillus firmus strain 37 in Bacillus subtilis WB800

Bacillusfirmus strain 37 produces the cyclomaltodextrin glucanotransferase (CGTase) enzyme and CGTase produces cyclodextrins (CDs) through a starch cyclization reaction. The strategy for the cloning and expression of recombinant CGTase is a potentially viable alternative for the economically viable production of CGTase for use in industrial processes. The present study used Bacillus subtilis WB800 as a bacterial expression host for the production of recombinant CGTase cloned from the CGTase gene of B. firmus strain 37. The CGTase gene was cloned in TOPO-TA ® plasmid, which was transformed in Escherichia coli DH5α. The subcloning was carried out with pWB980 plasmid and transformation in B. subtilis WB800. The 2xYT medium was the most suitable for the production of recombinant CGTase. The enzymatic activity of the crude extract of the recombinant CGTase of B. subtilis WB800 was 1.33 µmol β-CD/min/mL, or 7.4 times greater than the enzymatic activity of the crude extract of CGTase obtained from the wild strain. Following purification, the recombinant CGTase exhibited an enzymatic activity of 157.78 µmol β-CD/min/mL, while the activity of the CGTase from the wild strain was 9.54 µmol β-CD/min/mL. When optimal CDs production conditions for the CGTase from B. firmus strain 37 were used, it was observed that the catalytic properties of the CGTase enzymes were equivalent. The strategy for the cloning and expression of CGTase in B. subtilis WB800 was efficient, with the production of greater quantities of CGTase than with the wild strain, offering essential data for the large-scale production of the recombinant enzyme.Fil: Gimenez, Gabriela Gregolin. Universidad Estadual de Maringá; BrasilFil: Costa, Hernán. Universidad Nacional de Luján. Departamento de Ciencias Básicas. Area de Química Biológica; Argentina. Universidad Nacional de Luján. Instituto de Ecología y Desarrollo Sustentable. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Ecología y Desarrollo Sustentable; ArgentinaFil: de Lima Neto, Quirino Alves. Universidad Estadual de Maringá; BrasilFil: Fernandez, Maria Aparecida. Universidad Estadual de Maringá; BrasilFil: Ferrarotti, Susana Alicia. Universidad Nacional de Luján. Departamento de Ciencias Básicas. Area de Química Biológica; ArgentinaFil: Matioli, Graciette. Universidad Estadual de Maringá; Brasi

Vitamin D receptor gene polymorphisms are associated with leprosy in southern Brazil

Vitamin D, together with its nuclear receptor (VDR), plays an important role in modulating the immune response, decreasing the inflammatory process. Some polymorphisms of the VDR gene, such as BsmI (G>A rs1544410), ApaI (G>T rs7975232), and TaqI (T>C rs731236) could affect its stability and mRNA transcription activity, while FokI T>C (rs2228570) gives a truncated protein with three fewer amino acids and more efficiency in binding vitamin D. This study evaluated these four polymorphisms in the immunopathogenesis of leprosy in 404 patients and 432 control individuals without chronic or infectious disease in southern Brazil. When analyzing differences in the allele and genotype frequency of polymorphisms between patients (leprosy per se, multibacillary, and paucibacillary clinical forms) and controls, we found no statistically significant association. Regarding haplotype analysis, the bAt haplotype was associated with protection from leprosy per se (P = 0.004, OR = 0.34, CI = 0.16-0.71) and from the multibacillary clinical form (P = 0.005, OR = 0.30, CI = 0.13-0.70). In individuals aged 40 or more years, this haplotype has also showed protection against leprosy per se and multibacillary (OR = 0.26, CI = 0.09-0.76; OR = 0.26, CI = 0.07-0.78, respectively), while the BAt haplotype was a risk factor for leprosy per se in the same age group (OR = 1.34, CI = 1.04-1.73). In conclusion, despite having found no associations between the VDR gene polymorphisms with the development of leprosy, the haplotypes formed by the BsmI, ApaI, and TaqI polymorphisms were associated with leprosy per se and the multibacillary clinical form10CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQCOORDENAÇÃO DE APERFEIÇOAMENTO DE PESSOAL DE NÍVEL SUPERIOR - CAPESFUNDAÇÃO ARAUCÁRIA DE APOIO AO DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO DO ESTADO DO PARANÁ - FAsem informaçãosem informação1589/2017-CSDUE

Alternativas para práticas de microscopia no ensino fundamental: um estudo de caso

Introduction: Microscopy techniques are dynamic tools for the study of cytology in elementary school. Objective: To present a report on a report on the experiences lived from the use of two microscopy techniques. Methods: This study took place in May 2019, at Escola Municipal Mágylla Neto with students from two classes of the 7th year of Elementary School II. The study consisted of two practices: use of a digital optical microscope. In the second practice: assembly of a “home microscope” to check for live microorganisms in contaminated water. Results: The activity given after the practice, we analyzed the answers of 33 students. We verified a prevalence of 86% of correct answers, in front of 14% of questions answered wrongly. Despite being complementary, these data demonstrate the practices that allowed the assimilation of student learning. Conclusion: The practices allowed students to gain in-depth knowledge about something that only theory would make understanding more difficult.Introdução: Técnicas de microscopia são ferramentas dinâmicas para o estudo de citologia no ensino fundamental. Objetivo: Apresentar um relato sobre um relato sobre as experiências vivenciadas a partir da utilização de duas técnicas de microscopia. Métodos: Este estudo ocorreu no mês Maio de 2019, na Escola Municipal Mágylla Neto com alunos de duas turmas do 7º ano do Ensino Fundamental II. O estudo constituiu-se em duas práticas: utilização de um microscópio óptico digital. Na segunda prática: montagem de um “microscópio caseiro” para verificação de microrganismos vivos em água contaminada. Resultados: A atividade ministrada após a prática, analisamos as respostas de 33 alunos. Verificamos uma prevalência de 86% de acertos, diante 14% de questões respondidas erroneamente. Apesar de complementar, esses dados demonstram as práticas permitiram a assimilação da aprendizagem dos estudantes. Conclusão: As práticas permitiram aos alunos obterem um conhecimento aprofundado sobre algo que somente pela teoria tornaria mais difícil a compreensã