Detection of coinfection with Primate Erythroparvovirus 1 and arboviruses (DENV, CHIKV and ZIKV) in individuals with acute febrile illness in the state of Rio Grande do Norte in 2016.

BACKGROUND: Arthropod-borne viruses, known as arboviruses, pose substantial risks to global public health. Dengue (DENV), Chikungunya (CHIKV) and Zika (ZIKV) viruses stand out as significant concerns in Brazil and worldwide. Their overlapping clinical manifestations make accurate diagnosis a challenge, underscoring the need for reliable laboratory support. This study employs a comprehensive molecular diagnostic approach to track viral infections in individuals with acute febrile illness, a period marked by widespread outbreaks of DENV, CHIKV and ZIKV. METHODS: Between January and August 2016, we received a total of 713 serum samples obtained from individuals with acute febrile illness, previously tested for DENV, CHIKV or ZIKV, with initial negative results, from LACEN-NATAL. Of the total 713 samples, 667 were from females (354 of them pregnant) and 46 from males. Molecular diagnosis was conducted using the Multiplex RT-qPCR technique for simultaneous detection of DENV, CHIKV and ZIKV. Additionally, we performed differential diagnosis by RT-qPCR for other viruses of the Flavivirus, Alphavirus Enterovirus genera and qPCR for Primate Erythroparvovirus 1 (B19V) species, in accordance with Ministry of Health guidelines. RESULTS: Among the 713 cases, 78.2% tested positive for viral infections, including 48% with CHIKV viremia, 0.6% with DENV and 0.1% with ZIKV. Arboviral coinfections totaled 2.4%, including DENV-CHIKV (1.7%) and CHIKV-ZIKV (0.7%). Moreover, 8% exhibited B19V viremia. Simultaneous infections were identified in 17.5%, encompassing B19V-CHIKV (17.1%), B19V-DENV (0.1%), and B19V-ZIKV (0.3%) Triple infections were observed in 1.3% of cases with B19V-DENV-CHIKV (1%) and B19V-CHIKV-ZIKV (0.3%). CONCLUSION: Molecular testing demonstrated high efficacy in diagnosing prevalent arboviruses and detecting multiple coinfections. This approach helps to elucidate etiologies for symptomatic cases, especially during arbovirus outbreaks, and aids comprehensive surveillance. Our findings underscore the importance of monitoring co-circulating pathogens, such as B19V, with implications for clinical management, particularly in pregnant individuals. This study enhances our understanding of arbovirus epidemiology and reinforces the critical role of molecular diagnosis in disease surveillance and control.

Material genético do SARS-CoV-2 em superfícies de equipamentos utilizados na prática de atividades físicas / Genetic material of SARS-CoV-2 on the surface of equipment used in the practice of physical activities

A humanidade foi impactada por uma Pandemia que expôs a população ao contato com um vírus de elevado contágio e com o índice de letalidade alarmante. Este estudo objetivou avaliar a possibilidade da persistência de material genético do SARS-CoV-2 na superfície dos equipamentos de estabelecimentos de prática de atividades físicas indoor e outdoor. Foram coleta das amostras de equipamentos utilizados para a prática de exercícios físicos em cinco academias, cinco praças e entre os frequentadores desses ambientes. Aplicou-se a técnica RT-PCR para a detecção doRNA do SARS-CoV-2 e posterior análise dos resultadose foi detectada a existência de partículas de RNA viral do SARS-CoV-2 em 48,57% das amostras coletadas dos equipamentos das academias e 12,85% das amostras coletadas nas praças, evidenciando uma incidência menor em equipamentos utilizados em locais abertos em todas as áreas comparadas.Além disso, constatou-se que 35,7% dos participantes do estudo testaram positivo para COVID-19.Os casos positivos para COVID-19 detectados apresentaram sintomas classificados como levesa moderados e uma recuperação rápida.A presença de material genético nos equipamentos,por sua vez, leva-nos a perceber a importância da higienização adequada das superfícies, como forma de prevenção.

Humanity was impacted by a Pandemic that exposed the population to contact with a highly contagious virus with an alarming lethality rate. The present study aimed to evaluate the possibility of the persistence of genetic material from SARS-CoV-2 on the surface of equipment used to practice indoor and outdoor physical activities. A sample of equipment used for the practice of physical activity was collected in five gyms and five squares and among the regulars of these environments. The RT-PCR technique was applied to detect the RNA of SARS-CoV-2 and subsequent analysis of the results. The existence of SARS-CoV-2 viral RNA particles was detected in 48.57% of the samples collected from gym equipment and 12.85% of the samples collected in squares, evidencing a lower incidence in equipment used in open spaces in all areas compared and it was found that 35.7% of the study participants tested positive for COVID-19. The positive cases for COVID-19 detected, had symptoms classified as mild to moderate and a quick recovery. The presence of genetic material in the equipment, in turn, leads us to realize the importance of proper cleaning of surfaces, as a form of prevention.

Screening by high-throughput sequencing for pathogenic variants in cystic fibrosis: Benefit of introducing personalized therapies.

This short report documented cystic fibrosis transmembrane conductance regulator (CFTR) variants in 37 patients with cystic fibrosis (CF) in the Rio Grande do Norte region of Northeast Brazil. The high-throughput sequencing technology (HTS) genetic testing provided a definitive molecular diagnosis in 31 patients (83.8%). Among them, 25 patients' carriers of the c.1521_1523delCTT variant, categorized as a class 2 mutation, can be currently treated with CFTR modulator drugs. Five children aged 2-5 years could benefit from double lumacaftor/ivacaftor therapy, and 20 patients aged >6 years could be treated with the triple-combination elexacaftor/tezacaftor/ivacaftor therapy. Thus, the identification of pathogenic variants associated with the development of this disease allows for the introduction of therapy with CFTR modulators that favour better patient management.

Severe Acute Respiratory Syndrome Coronavirus 2 P.2 Lineage Associated with Reinfection Case, Brazil, June-October 2020.

A 37-year-old healthcare worker from the northeastern region of Brazil experienced 2 clinical episodes of coronavirus disease. Infection with severe acute respiratory syndrome coronavirus 2 was confirmed by reverse transcription PCR in samples collected 116 days apart. Whole-genome sequencing revealed that the 2 infections were caused by the most prevalent lineage in Brazil, B.1.1.33, and the emerging lineage P.2. The first infection occurred in June 2020; Bayesian analysis suggests reinfection at some point during September 14-October 11, 2020, a few days before the second episode of coronavirus disease. Of note, P.2 corresponds to an emergent viral lineage in Brazil that contains the mutation E484K in the spike protein. The P.2 lineage was initially detected in the state of Rio de Janeiro, and since then it has been found throughout the country. Our findings suggest not only a reinfection case but also geographic dissemination of the emerging Brazil clade P.2.

A Potential SARS-CoV-2 Variant of Interest (VOI) Harboring Mutation E484K in the Spike Protein Was Identified within Lineage B.1.1.33 Circulating in Brazil.

The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) epidemic in Brazil was dominated by two lineages designated as B.1.1.28 and B.1.1.33. The two SARS-CoV-2 variants harboring mutations at the receptor-binding domain of the Spike (S) protein, designated as lineages P.1 and P.2, evolved from lineage B.1.1.28 and are rapidly spreading in Brazil. Lineage P.1 is considered a Variant of Concern (VOC) because of the presence of multiple mutations in the S protein (including K417T, E484K, N501Y), while lineage P.2 only harbors mutation S:E484K and is considered a Variant of Interest (VOI). On the other hand, epidemiologically relevant B.1.1.33 deriving lineages have not been described so far. Here we report the identification of a new SARS-CoV-2 VOI within lineage B.1.1.33 that also harbors mutation S:E484K and was detected in Brazil between November 2020 and February 2021. This VOI displayed four non-synonymous lineage-defining mutations (NSP3:A1711V, NSP6:F36L, S:E484K, and NS7b:E33A) and was designated as lineage N.9. The VOI N.9 probably emerged in August 2020 and has spread across different Brazilian states from the Southeast, South, North, and Northeast regions.

Chikungunya virus ECSA lineage reintroduction in the northeasternmost region of Brazil.

The Northeast region of Brazil registered the second-highest incidence proportion of Chikungunya fever in 2019. In that year, an outbreak consisting of patients presenting with febrile disease associated with joint pain was reported by the public primary health care service in the city of Natal, in the state of Rio Grande do Norte, in March 2019. At first, the aetiological agent of the disease was undetermined. Since much is still unknown about chikungunya virus' (CHIKV) genomic diversity and evolutionary history in this northeasternmost state, we used a combination of portable whole-genome sequencing, molecular clock, and epidemiological analyses that revealed the reintroduction of the CHIKV East-Central-South-African (ECSA) lineage into Rio Grande do Norte. We estimated that the CHIKV ECSA lineage was first introduced into Rio Grande do Norte in early June 2014, while the 2019 outbreak clade diverged around April 2018, during a period of increased Chikungunya incidence in the Southeast region, which might have acted as a source of virus dispersion towards the Northeast region. Together, these results confirm that the ECSA lineage continues to spread across the country through interregional importation events, likely mediated by human mobility.

A Potential SARS-CoV-2 Variant of Interest (VOI) Harboring Mutation E484K in the Spike Protein Was Identified within Lineage B 1 1 33 Circulating in Brazil

The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) epidemic in Brazil was dominated by two lineages designated as B.1.1.28 and B.1.1.33. The two SARS-CoV-2 variants harboring mutations at the receptor-binding domain of the Spike (S) protein, designated as lineages P.1 and P.2, evolved from lineage B.1.1.28 and are rapidly spreading in Brazil. Lineage P.1 is considered a Variant of Concern (VOC) because of the presence of multiple mutations in the S protein (including K417T, E484K, N501Y), while lineage P.2 only harbors mutation S:E484K and is considered a Variant of Interest (VOI). On the other hand, epidemiologically relevant B.1.1.33 deriving lineages have not been described so far. Here we report the identification of a new SARS-CoV-2 VOI within lineage B.1.1.33 that also harbors mutation S:E484K and was detected in Brazil between November 2020 and February 2021. This VOI displayed four non-synonymous lineage-defining mutations (NSP3:A1711V, NSP6:F36L, S:E484K, and NS7b:E33A) and was designated as lineage N.9. The VOI N.9 probably emerged in August 2020 and has spread across different Brazilian states from the Southeast, South, North, and Northeast regions.

IRF6 polymorphisms in Brazilian patients with non-syndromic cleft lip with or without palate / Polimorfismos do gene IRF6 em pacientes brasileiros com fenda labial não sindrômica com ou sem fenda palatina

Abstract Introduction: Non-syndromic orofacial clefts have a complex etiology due to the contribution from both genetic and environmental risk factors, as well as the interaction between them. Among the more than 15 susceptibility loci for non-syndromic orofacial clefts with considerable statistical and biological support, the IRF6 is the most validated gene by the majority of studies. Nonetheless, in genetically heterogeneous populations such as Brazilian, the confirmation of association between non-syndromic orofacial clefts and IRF6 common variants is not a consolidated fact and unrecognized IRF6 variants are poorly investigated. Objective: The aim of this study was to investigate the association of IRF6 polymorphisms with non-syndromic orofacial clefts development in a population from northeast Brazil. Methods: Blood samples of 186 non-syndromic orofacial clefts patients and 182 controls from Rio Grande do Norte, Brazil, were obtained to analyze IRF6 polymorphisms (rs2235371, rs642961, rs2236907, rs861019, and rs1044516) by real-time polymerase chain reaction. Non-syndromic orofacial clefts patients were classified in cleft lip and palate, cleft palate only and cleft lip only groups. Results: The genotype and allele frequencies of single nucleotide polymorphism rs2235371 in IRF6 showed significant differences in patients with cleft palate when compared to the controls, whereas no association was shown between rs642961, rs2236907, rs861019, and rs1044516 and non-syndromic orofacial clefts. Conclusion: The association found between rs2235371 and isolated cleft palate should be interpreted with caution due to the low number of individuals investigated, and more studies with larger sample size are needed to confirm these association. In addition, there is a lack of association of the rs642961, rs2236907 and rs861019 polymorphisms with non-syndromic orofacial clefts susceptibility.

Resumo Introdução: As fendas orofaciais não sindrômicas possuem uma etiologia complexa devido à contribuição de fatores de risco genéticos e ambientais, assim como a interação entre eles. Dentre os mais de 15loci de susceptibilidade para as fendas orofaciais não sindrômicas com considerável suporte estatístico e biológico, o IRF6 é o gene mais validado pela maioria dos estudos. Apesar disso, em populações geneticamente heterogêneas como a brasileira, a confirmação da associação entre as fendas orofaciais não sindrômicas e as variantes mais comuns do IRF6 ainda não é um fato consolidado e outras variantes não tão conhecidas IRF6 são pouco investigadas. Objetivo: O objetivo deste estudo foi investigar a associação de variados polimorfismos do IRF6 com o desenvolvimento das fendas orofaciais não sindrômicas em uma população do nordeste do Brasil. Método: Amostras de sangue de 186 pacientes com fendas orofaciais não sindrômicas e 182 controles do estado do Rio Grande do Norte, Brasil, foram obtidas para analisar os polimorfismos do IRF6 (rs2235371, rs642961, rs2236907, rs861019 e rs1044516) por reação em cadeia da polimerase em tempo real. Os pacientes com fendas orofaciais não sindrômicas foram classificados em fenda labiopalatina, fenda palatina isolada e fenda labial isolada. Resultados: As frequências genotípica e alélica do polimorfismo de único nucleotídeo rs2235371 no IRF6 mostraram-se significativamente diferentes em pacientes com fenda palatina isolada quando comparadas às dos controles, enquanto que nenhuma associação foi encontrada entre rs642961, rs2236907, rs861019 e rs1044516 e risco para o desenvolvimento das fendas orofaciais não sindrômicas. Conclusão: A associação encontrada entre rs2235371 e fenda palatina isolada deve ser interpretada com cautela devido ao baixo número de indivíduos investigados, sendo necessários mais estudos com um tamanho amostral maior para confirmar essa associação. Além disso, não foram encontradas associações significativas entre os demais polimorfismos do IRF6 rs642961, rs2236907, rs861019 e rs1044516 e a susceptibilidade às fendas orofaciais não sindrômicas.

IRF6 polymorphisms in Brazilian patients with non-syndromic cleft lip with or without palate.

INTRODUCTION: Non-syndromic orofacial clefts have a complex etiology due to the contribution from both genetic and environmental risk factors, as well as the interaction between them. Among the more than 15 susceptibility loci for non-syndromic orofacial clefts with considerable statistical and biological support, the IRF6 is the most validated gene by the majority of studies. Nonetheless, in genetically heterogeneous populations such as Brazilian, the confirmation of association between non-syndromic orofacial clefts and IRF6 common variants is not a consolidated fact and unrecognized IRF6 variants are poorly investigated. OBJECTIVE: The aim of this study was to investigate the association of IRF6 polymorphisms with non-syndromic orofacial clefts development in a population from northeast Brazil. METHODS: Blood samples of 186 non-syndromic orofacial clefts patients and 182 controls from Rio Grande do Norte, Brazil, were obtained to analyze IRF6 polymorphisms (rs2235371, rs642961, rs2236907, rs861019, and rs1044516) by real-time polymerase chain reaction. Non-syndromic orofacial clefts patients were classified in cleft lip and palate, cleft palate only and cleft lip only groups. RESULTS: The genotype and allele frequencies of single nucleotide polymorphism rs2235371 in IRF6 showed significant differences in patients with cleft palate when compared to the controls, whereas no association was shown between rs642961, rs2236907, rs861019, and rs1044516 and non-syndromic orofacial clefts. CONCLUSION: The association found between rs2235371 and isolated cleft palate should be interpreted with caution due to the low number of individuals investigated, and more studies with larger sample size are needed to confirm these association. In addition, there is a lack of association of the rs642961, rs2236907 and rs861019 polymorphisms with non-syndromic orofacial clefts susceptibility.

Síndrome Respiratória Aguda Grave e a COVID-19 (SARS-Cov-2): uma revisão narrativa / Severe acute respiratory syndrome and covid-19 (sars-cov-2): a narrative review

Objetivo: Analisar a produção científica acerca da atual pandemia do novo coronavírus, destacando aspectos referentes às características do vírus, bem como a epidemiologia, o diagnóstico e tratamento da COVID-19 no intuito de fornecer informações aos profissionais de saúde e à comunidade científica. Método: Revisão narrativa, com busca nas bases de dados Scielo, Medline, Lilacs e Pubmed, referente ao período de fevereiro a maio de 2020, utilizando os descritores: "infecções por coronavírus" or "coronavirus infections", "COVID-19", "betacoronavirus", "Síndrome Respiratória Aguda Grave", "SARS-Cov-2", "diagnóstico", "diagnosis". Resultados: Foram evidenciadas três categorias temáticas: aspectos referentes ao vírus; a epidemiologia e a clínica da doença; e aspectos do diagnóstico e tratamento. Conclusão: O Coronavírus sofreu mutações desde o início da pandemia, mas não se sabe se estas mutações afetaram seu poder de infecciosidade. Em diferentes partes do mundo a epidemiologia da COVID-19 foi diferente. Com relação ao tratamento, a droga que tem se mostrado mais promissora até o momento é o remdesivir. Sem uma vacina eficaz, a maior arma que dispomos é o isolamento social. (AU)

Objective: To analyze a scientific production on the current pandemic of new coronaviruses, highlighting aspects related to virus resources, as well as the epidemiology, diagnosis and treatment of COVID-19 in order to provide information to health professionals and the scientific community. Method: Narrative review, searching the Scielo, Medline, Lilacs and Pubmed databases, for the period from February to May 2020, using the keywords: "coronavirus infections" or "coronavirus infections", "COVID-19", "Betacoronavirus", "Severe Acute Respiratory Syndrome", "SARS-Cov-2", "diagnosis", "diagnosis". Results: Three thematic categories were highlighted: aspects related to the virus; an epidemiology and disease clinic; and aspects of diagnosis and treatment. Conclusion: The Coronavirus has mutated since the beginning of the pandemic, but it is not known whether these mutations have affected its infectiousness. In different parts of the world the epidemiology of COVID-19 was different. Regarding treatment, a drug that has been shown to be more promising until recovery is needed. Without an effective vaccine, the biggest weapon that affects social isolation. (AU)

Objetivo: analizar una producción científica sobre la pandemia actual de nuevos coronavirus, destacando aspectos relacionados con los recursos del virus, así como la epidemiología, diagnóstico y tratamiento de COVID-19 para proporcionar información a los profesionales de la salud y la comunidad científica. Método: Revisión narrativa, buscando en las bases de datos Scielo, Medline, Lilacs y Pubmed, para el período de febrero a mayo de 2020, utilizando los descriptores: "infecciones por coronavirus" o "infecciones por coronavirus", "COVID-19", "Betacoronavirus", "Síndrome respiratorio agudo severo", "SARS-Cov-2", "diagnóstico", "diagnóstico". Resultados: se destacaron tres categorías temáticas: aspectos relacionados con el virus; una clínica de epidemiología y enfermedad; y aspectos de diagnóstico y tratamiento. Conclusión: el coronavirus ha mutado desde el comienzo de la pandemia, pero no se sabe si estas mutaciones han afectado su infecciosidad. En diferentes partes del mundo, la epidemiología de COVID-19 fue diferente. Con respecto al tratamiento, un medicamento que ha demostrado ser más prometedor hasta que se necesita recuperación. Sin una vacuna efectiva, el arma más grande que afecta el aislamiento social. (AU)

Estratégia de enfrentamento de diagnóstico laboratorial da COVID-19 no estado da Paraíba / Covid-19 laboratory diagnosis coping strategy in the state of Paraíba

Objetivo: Relatar a experiência de uma equipe multiprofissional na implementação de um laboratório, para realização de testes para diagnóstico precoce de infecção pelo Coronavírus da Síndrome Respiratória Aguda Grave 2 (SARS-CoV-2). Método: Estudo descritivo, tipo relato de experiência, desenvolvido numa Escola Técnica de Saúde Federal de João Pessoa, Paraíba. Resultados: A experiência ocorreu em três etapas: na primeira etapa foi realizada a organização do laboratório; na segunda etapa a descrição do funcionamento deste, com acolhimento e análise das amostras da nasofaringe e orofaringe dos indivíduos e profissionais da saúde; e, na terceira etapa que constitui a investigação dos indivíduos e avaliação de profissionais da saúde com risco para a doença do coronavírus 2019 (COVID-19), por meio do mapeamento das unidades assistenciais para o SARS-CoV-2 e outros vírus respiratórios, com o intuito de estabelecer o perfil epidemiológico da população exposta. Conclusão: A estratégia desenvolvida pela Escola Técnica de Saúde em apoiar o estado da Paraíba nas ações de controle e mitigação da pandemia da COVID-19, mais especificamente no diagnóstico da doença, permitiu grande avanço do ponto de vista do conhecimento científico e contribuição na geração de informação, capacitação, produtos, processos ou serviços especializados. (AU)

Objective: To report the experience of a multidisciplinary team in the implementation of a laboratory, to carry out tests for the early diagnosis of infection by the Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Method: Descriptive study, type of experience report, developed at a Federal Health Technical School in João Pessoa, Paraíba. Results: The experience took place in three stages: in the first stage, the laboratory was organized; in the second stage the description of its functioning, with reception and analysis of the nasopharynx and oropharynx samples of individuals and health professionals; and, in the third stage that constitutes the investigation of individuals and the evaluation of health professionals at risk for coronavirus disease-2019 (COVID-19), through the mapping of care units for SARS-CoV-2 and other respiratory viruses, in order to establish the epidemiological profile of the exposed population. Conclusion: A strategy developed by the Technical School of Health in support of Paraíba state, in the actions to control and mitigate the pandemic of COVID-19, more specific in the diagnosis of the disease, allowed a great advance in terms of scientific knowledge and research scientific information, training, products, processes or specialized services. (AU)

Objetivo: Informar la experiencia de un equipo multidisciplinario en la implementación de un laboratorio, para llevar a cabo pruebas para el diagnóstico precoz de infección por el Coronavirus del Síndrome Respiratorio Agudo Grave 2 (SARS-CoV-2). Método: Estudio descriptivo, tipo de informe de experiencia, desarrollado en una Escuela Técnica Federal de Salud en João Pessoa, Paraíba. Resultados: El experimento se realizó en tres etapas: en la primera etapa, se organizó el laboratorio; en la segunda etapa la descripción de su funcionamiento, con recepción y análisis de muestras de individuos y profesionales de la salud de la nasofaringe y la orofaringe; y, en la tercera etapa que constituye la investigación de individuos y la evaluación de profesionales de la salud en riesgo de contraer la enfermedad por coronavirus 2019 (COVID-19), a través del mapeo de unidades de atención para SARS-CoV-2 y otros virus respiratorios, para establecer el perfil epidemiológico de la población expuesta. Conclusión: La estrategia desarrollada por la Escuela Técnica de Salud para apoyar al estado de Paraíba en las acciones para controlar y mitigar la pandemia de COVID-19, más específicamente en el diagnóstico de la enfermedad, permitió un gran progreso desde el punto de vista del conocimiento científico y la contribución a la generación. información, formación, productos, procesos o servicios especializados. (AU)

Risk factors and comorbidities in Brazilian patients with orofacial clefts.

Considering that environmental risk factors substantially contribute to the etiology of orofacial clefts and that knowledge about the characteristics and comorbidities associated with oral clefts is fundamental to promoting better quality of life, this study aimed to describe the risk factors, main characteristics, and comorbidities of a group of patients with cleft lip and/or cleft palate (CL/P) from Rio Grande do Norte (RN), Brazil. Data were obtained from 173 patients with CL/P using a form from the Brazilian database on Orofacial Clefts. Most patients were male with cleft lip and palate and had a normal size and weight at birth; presented few neonatal intercurrent events; and had anemia and respiratory and cardiovascular diseases as main associated comorbidities. They also required timely surgical rehabilitation and multidisciplinary care to stimulate their neuropsychomotor development. In addition, a high frequency of familial recurrence and of parental consanguinity was evidenced in the studied population, especially for the cleft lip and cleft palate type. Other relevant findings were the considerable maternal exposure to alcohol, infections, smoking, and hypertension, as well as low supplementation with vitamins and minerals and deliberate consumption of analgesics, antibiotics, and antihypertensives during pregnancy. Characterization of the CL/P patient profile is essential for the planning of health services and integration among the health professionals involved in the diagnosis and treatment of these malformations. Our results reinforce the need for additional research to confirm the association between environmental factors and the development of orofacial clefts.

Application of high-resolution array platform for genome-wide copy number variation analysis in patients with nonsyndromic cleft lip and palate.

BACKGROUND: Although more than 14 loci may be involved in the development of nonsyndromic cleft lip and palate (NSCLP), the etiology has not been fully elucidated due to genetic and environmental risk factor interactions. Despite advances in identifying genes associated with the NSCLP development using traditional genetic mapping strategies of candidate genes, genome-wide studies, and epidemiologic and linkage analysis, microarray techniques have become important complementary tools in the search for potential causative oral clefts genes in genetic studies. Microarray hybridization enables scanning of the whole genome and detecting copy number variants (CNVs). Although common benign CNVs are often smaller, with sizes smaller than 20 kb, here we reveal small exonic CNVs based on the importance of the encompassed genes in cleft lip and palate phenotype. METHODS: Microarray hybridization analysis was performed in 15 individuals with NSCLP. RESULTS: We identified 11 exonic CNVs affecting at least one exon of the candidate genes. Thirteen candidate genes (COL11A1-1p21; IRF6-1q32.3; MSX1-4p16.2; TERT-5p15.33; MIR4457-5p15.33; CLPTM1L-5p15.33; ESR1-6q25.1; GLI3-7p13; FGFR-8p11.23; TBX1-22q11.21; OFD-Xp22; PHF8-Xp11.22; and FLNA-Xq28) overlapped with the CNVs identified. CONCLUSIONS: Considering the importance to NSCLP, the microdeletions that encompass MSX1, microduplications over TERT, MIR4457, CLPTM1L, and microduplication of PHF8 have been identified as small CNVs related to sequence variants associated with oral clefts susceptibility. Our findings represent a preliminary study on the clinical significance of small CNVs and their relationship with genes implicated in NSCLP.

Risk factors and comorbidities in Brazilian patients with orofacial clefts

Abstract: Considering that environmental risk factors substantially contribute to the etiology of orofacial clefts and that knowledge about the characteristics and comorbidities associated with oral clefts is fundamental to promoting better quality of life, this study aimed to describe the risk factors, main characteristics, and comorbidities of a group of patients with cleft lip and/or cleft palate (CL/P) from Rio Grande do Norte (RN), Brazil. Data were obtained from 173 patients with CL/P using a form from the Brazilian database on Orofacial Clefts. Most patients were male with cleft lip and palate and had a normal size and weight at birth; presented few neonatal intercurrent events; and had anemia and respiratory and cardiovascular diseases as main associated comorbidities. They also required timely surgical rehabilitation and multidisciplinary care to stimulate their neuropsychomotor development. In addition, a high frequency of familial recurrence and of parental consanguinity was evidenced in the studied population, especially for the cleft lip and cleft palate type. Other relevant findings were the considerable maternal exposure to alcohol, infections, smoking, and hypertension, as well as low supplementation with vitamins and minerals and deliberate consumption of analgesics, antibiotics, and antihypertensives during pregnancy. Characterization of the CL/P patient profile is essential for the planning of health services and integration among the health professionals involved in the diagnosis and treatment of these malformations. Our results reinforce the need for additional research to confirm the association between environmental factors and the development of orofacial clefts.

Anabolic Effect of Insulin Therapy on the Bone: Osteoprotegerin and Osteocalcin Up-Regulation in Streptozotocin-Induced Diabetic Rats.

Type 1 diabetes mellitus (T1DM) is associated with several skeletal alterations, particularly in conditions of poor glycaemic control. Insulin therapy is the major conservative treatment for T1DM; however, the effects of this hormone on bone markers of T1DM rats are limited, and the regulatory mechanisms remain elusive. Therefore, the evaluation of molecular and non-molecular parameters in a chronic animal model of T1DM-induced bone loss, treated with and without insulin, may help in elucidating the insulin mechanisms. Male Wistar rats were assigned into three groups: control, T1DM (T1DM rats induced with streptozotocin [STZ] at 40 mg/kg intravenously) and T1DM plus insulin therapy (T1DMI). After 8 weeks, we evaluated the serum biochemical, tibia histomorphometric and biomechanical parameters, as well as the gene expression of the receptor activator of nuclear factor kappa-B ligand (RANKL), osteoprotegerin (OPG) and osteocalcin (OC) of femur mRNA. Compared with T1DM, the T1DMI group showed less bone loss, which was revealed by the increased trabecular width (TbWi, p < 0.001) and trabecular bone area (BAr, p < 0.01), reduced trabecular separation (TbSp, p < 0.01) and increased Young's modulus (p < 0.05). Moreover, molecular analyses indicated that the expression of OPG and OC was up-regulated (p < 0.001 and p < 0.05, respectively). In summary, the up-regulation of OPG and OC in the T1DMI group supports an anabolic effect of insulin, which was demonstrated by the maintenance of bone architecture and flexibility. These results suggest that insulin therapy may prevent T1DM-induced bone loss via the effects on the bone formation.

Relationship between glycemic control and OPG gene polymorphisms with lower bone mineral density in patients with type 1 Diabetes mellitus

ABSTRACT The aim of the present study was to investigate the bone mineral density (BMD) of patients with type 1 Diabetes mellitus (T1DM). We also assessed the association between osteoprotegerin (OPG) genetic polymorphisms and BMD. Genotyping was performed for 1181G>C and 163A>G OPG polymorphisms by allelic discrimination in 119 patients with T1DM and 161 normoglycemic (NG) individuals, aged 6 to 20 years old. Glycemic control, serum parameters of bone metabolism and BMD were evaluated. T1DM patients showed low BMD, poor glycemic control and decreased total calcium values when compared to controls (p < 0.05). For all the polymorphisms studied, the genotype and allele frequencies in patients with T1DM were not significantly different from the controls. In patients with T1DM, carriers of OPG 1181CC showed higher concentrations of ionized calcium compared to patients with GG+GC genotypes. These results suggest that low BMD is associated with poor glycemic control in T1DM. Despite the lack of a detected association between OPG polymorphisms and BMD in these patients, the increased ionized calcium in those carrying OPG 1181CC suggests a possible increase in osteoclastogenesis, a conclusion that may be supported by the lower BMD observed in these subjects.

Protection against T1DM-Induced Bone Loss by Zinc Supplementation: Biomechanical, Histomorphometric, and Molecular Analyses in STZ-Induced Diabetic Rats.

Several studies have established an association between diabetes and alterations in bone metabolism; however, the underlying mechanism is not well established. Although zinc is recognized as a potential preventive agent against diabetes-induced bone loss, there is no evidence demonstrating its effect in chronic diabetic conditions. This study evaluated the effects of zinc supplementation in a chronic (90 days) type 1 diabetes-induced bone-loss model. Male Wistar rats were distributed in three groups: control, type 1 diabetes mellitus (T1DM), and T1DM plus zinc supplementation (T1DMS). Serum biochemical analysis; tibia histomorphometric, biomechanical, and collagen-content analyses; and femur mRNA expression were evaluated. Relative to T1DM, the zinc-supplemented group showed increased histomorphometric parameters such as TbWi and BAr and decreased TbSp, increased biomechanical parameters (maximum load, stiffness, ultimate strain, and Young's modulus), and increased type I collagen content. Interestingly, similar values for these parameters were observed between the T1DMS and control groups. These results demonstrate the protective effect of zinc on the maintenance of bone strength and flexibility. In addition, downregulation of OPG, COL1A, and MMP-9 genes was observed in T1DMS, and the anabolic effects of zinc were evidenced by increased OC expression and serum ALP activity, both related to osteoblastogenesis, demonstrating a positive effect on bone formation. In contrast, T1DM showed excessive bone loss, observed through reduced histomorphometric and biomechanical parameters, characterizing diabetes-associated bone loss. The bone loss was also observed through upregulation of OPG, COL1A, and MMP-9 genes. In conclusion, zinc showed a positive effect on the maintenance of bone architecture and biomechanical parameters. Indeed, OC upregulation and control of expression of OPG, COL1A, and MMP-9 mRNAs, even in chronic hyperglycemia, support an anabolic and protective effect of zinc under chronic diabetic conditions. Furthermore, these results indicate that zinc supplementation could act as a complementary therapy in chronic T1DM.

Association of polymorphisms in IL6 gene promoter region with type 1 diabetes and increased albumin-to-creatinine ratio.

BACKGROUND: Pro-inflammatory cytokines, such as interleukin-6 (IL-6), have been considered as key factors in type 1 diabetes mellitus (T1DM) and diabetic nephropathy, thus, our aim was to investigate the association of IL6-174G>C (rs1800795) and -634C>G (rs1800796) polymorphisms with T1DM susceptibility and diabetic nephropathy. METHODS: These polymorphisms were analyzed in 144 children and adolescents with T1DM and 173 normoglycemic control subjects. Glycemic control, laboratory parameters of kidney function and serum lipids were evaluated. By studying only T1DM patients, we evaluated the polymorphisms associated with relevant biochemical parameters in various genetic models. RESULTS: Type 1 diabetes mellitus patients showed poor glycemic control and albumin-to-creatinine ratio, total cholesterol and LDL-cholesterol levels increased when compared with normoglycemic subjects (p < 0.001, p = 0.004 and p < 0.001, respectively). IL6-174C allele was associated with an increased risk of developing T1DM (OR = 1.53, CI = 1.01-2.31, p = 0.044). In the T1DM group, IL6-174CC carriers showed higher concentrations of glycated hemoglobin (p = 0.029), albumin-to-creatinine ratio (p = 0.021), total cholesterol (p = 0.010), and LDL-cholesterol (p = 0.002), when compared with GG+GC carriers. No association was found for the IL6-634C>G polymorphism. CONCLUSIONS: These results suggest that IL6-174G>C may contribute to T1DM and increased albumin-to-creatinine ratio as well as to poor glycemic control and hyperlipidemia.

[Effect of tamoxifen on plasma proteins in diabetes mellitus type 1]. / Efeito do tamoxifeno no perfil das proteínas plasmáticas em condição de diabetes mellitus tipo 1.

PURPOSE: Considering that important scientific advances have been obtained through studies based on experimental Diabetes mellitus, and that tamoxifen action in humans remains unknown, the aim of the present work is to follow the modifications promoted by diabetes and tamoxifen in the electrophoretic profile of plasmatic proteins. METHODS: It was used 27 Wistar female rats (180-250 body weight), randomicaly divided into five groups: C1 (n = 3, received vehicle), C2 (n = 3, no treatment), T (n =5, treated with tamoxifen, 0.3mg/Kg/day), D (n = 8, experimental diabetes by estreptozotocin, 45mg/Kg and DT (n = 8, diabetic treated with tamoxifen). The electrophoresis was accomplished in cellulose acetate. pH 8.6-8.8, TECNOW chamber, and the strains were stained by Ponceau S. The total proteins were determined by the Biuret method (Labtest). Proteinograms were obtained in densitometer BioSystems BTS-235. RESULTS: Albumin decreased progressively in the groups T, D and DT; a1 fraction increased in groups T and DT; a2 fraction increased in groups T and D, including a synergic effect in group DT; a fraction increased in groups T and D; a fraction increased in groups T, D and DT. CONCLUSIONS: The results indicate an acute phase resposta, with synergic effect of tamoxifen and diabetes, suggesting a probable hepatic lesion.

Efeito do tamoxifeno no perfil das proteínas plasmáticas em condição de diabetes mellitus tipo 1 / Effect of tamoxifen on plasma proteins in diabetes mellitus type 1

OBJETIVO: Considerando-se que importantes avanços científicos têm sido obtidos através de estudos com Diabetes mellitus experimental, e que a ação do tamoxifeno em humanos permanece obscura, o presente trabalho objetiva acompanhar as modificações promovidas pelo diabetes e tamoxifeno no perfil eletroforético das proteínas plasmáticas. MÉTODOS: Foram utilizados 27 ratos fêmeas Wistar (180-220g peso corporal), divididos randomicamente em 5 grupos: C1 (n=3, receberam veículo), C2 (n=3, sem tratamento), T (n=5, tratados com tamoxifeno, 0,3mg/kg/dia), D (n=8, diabéticos experimentais por estreptozotocina, 45mg/Kg) e DT (n=8, diabéticos tratados com tamoxifeno). A eletroforese foi realizada em acetato de celulose, pH 8,6-8,8, cuba TECNOW, e as fitas foram coradas em Ponceau S. As proteínas totais foram determinadas pelo método do Biureto (Kit Labtest). Os proteinogramas foram obtidos em densitômetro BioSystems BTS-235. RESULTADOS: Albumina diminuiu progressivamente nos grupos T, D e DT; a fração a1 aumentou nos grupos T e DT; a fração a2 aumentou nos grupos T e D, havendo efeito aditivo no grupo DT; a fração b aumentou nos grupos T e D; a fração g aumentou nos grupos T, D e DT. CONCLUSÃO: Os resultados indicam uma resposta de fase aguda, com efeito aditivo do tamoxifeno e diabetes, sugerindo uma provável lesão hepática.