Association between genetic variants in TREM1, CXCL10, IL4, CXCL8 and TLR7 genes with the occurrence of congenital Zika syndrome and severe microcephaly.

Congenital Zika syndrome (CZS) is a cluster of malformations induced by Zika virus (ZIKV) infection and the underline mechanisms involved in its occurrence are yet not fully understood. Along with epidemiological and environmental factors, the genetic host factors are suggested as important to the CZS occurrence and development, however, few studies have evaluated this. This study enrolled a total of 245 individuals in a case-control association study compound a cohort of high specific interest constituted by 75 mothers who had delivered CZS infants, their 76 infants, and 47 mothers that had delivered healthy infants, and their 47 infants. Sixteen single-nucleotide polymorphisms on TREM1, CXCL10, IL4, CXCL8, TLR3, TLR7, IFNR1, CXCR1, IL10, CCR2 and CCR5 genes were genotyped to investigate their association as risk factors to CZS. The results show an association between C allele at TREM1 rs2234246 and C allele at IL4 rs224325 in mothers infected with ZIKV during pregnancy, with the increased susceptibility to CZS occurrence in their infants and the SNP CXCL8 rs4073 and the G allele at CXCL10 rs4508917 with presence of CZS microcephaly in the infants. Furthermore, the T allele at CXCL8 rs4073 and TRL7 rs179008 SNPs were associated with the severity of microcephaly in children with CZS. These results suggest that these polymorphisms in genes of innate immune responses addressed here are associated to increased risk of occurrence and severity of CZS in pregnant mothers infected with ZIKV and their CZS infants.

Recurrent COVID-19 including evidence of reinfection and enhanced severity in thirty Brazilian healthcare workers.

BACKGROUND: There is growing concern about individuals reported to suffer repeat COVID-19 disease episodes, these in a small number of cases characterised as de novo infections with distinct sequences, indicative of insufficient protective immunity even in the short term. METHODS: Observational case series and case-control studies reporting 33 cases of recurrent, symptomatic, qRT-PCR positive COVID-19. Recurrent disease was defined as symptomatic recurrence after symptom-free clinical recovery, with release from isolation >14 days from the beginning of symptoms confirmed by qRT-PCR. The case control study-design compared this group of patients with a control group of 62 patients randomly selected from the same COVID-19 database. RESULTS: Of 33 recurrent COVID-19 patients, 26 were female and 30 were HCW. Mean time to recurrence was 50.5 days which was associated with being a HCW (OR 36.4 (p <0.0001)), and blood type A (OR 4.8 (p = 0.002)). SARS-CoV-2 antibodies were signifcantly lower in recurrent patients after initial COVID-19  (2.4 ±â€¯0.610; p<0.0001) and after recurrence (6.4 ±â€¯11.34; p = 0.007).  Virus genome sequencing identified reinfection by a different isolate in one patient. CONCLUSIONS: This is the first detailed case series showing COVID-19 recurrence with qRT-PCR positivity. For one individual detection of phylogenetically distinct genomic sequences in the first and second episodes confirmed bona fide renfection, but in most cases the data do not formally distinguish between reinfection and re-emergence of a chronic infection reservoir. These episodes were significantly associated with reduced Ab response during initial disease and argue the need for ongoing vigilance without an assumption of protection after a first episode.

Strong CD4 T Cell Responses to Zika Virus Antigens in a Cohort of Dengue Virus Immune Mothers of Congenital Zika Virus Syndrome Infants.

Background: There is an urgent need to understand the complex relationship between cross-reactive anti-viral immunity, disease susceptibility, and severity in the face of differential exposure to related, circulating Flaviviruses. Co-exposure to Dengue virus and Zika virus in Brazil is a case in point. A devastating aspect of the 2015-2016 South American Zika outbreak was the dramatic increase in numbers of infants born with microcephaly to mothers exposed to Zika virus during pregnancy. It has been proposed that this is more likely to ensue from Zika infection in women lacking cross-protective Dengue immunity. In this case series we measure the prevalence of Dengue immunity in a cohort of mothers exposed to Zika virus during pregnancy in the 2015-2016 Zika outbreak that gave birth to an infant affected by microcephaly and explore their adaptive immunity to Zika virus. Results: Fifty women from Sergipe, Brazil who gave birth to infants with microcephaly following Zika virus exposure during the 2015-16 outbreak were tested for serological evidence of Dengue exposure and IFNγ ELISpot spot forming cell (SFC) response to Zika virus. The majority (46/50) demonstrated Dengue immunity. IFNγ ELISpot responses to Zika virus antigens showed the following hierarchy: Env>NS1>NS3>C protein. Twenty T cell epitopes from Zika virus Env were identified. Responses to Zika virus antigens Env and NS1 were polyfunctional with cells making IFNγ, TNFα, IL-4, IL-13, and IL-10. In contrast, responses to NS5 only produced the immune regulatory TGFß1 cytokine. There were SFC responses against Zika virus Env (1-20) and variant peptide sequences from West Nile virus, Dengue virus 1-4 and Yellow Fever virus. Conclusion: Almost all the women in our study showed serological evidence of Dengue immunity, suggesting that microcephaly can occur in DENV immune mothers. T cell immunity to Zika virus showed a multifunctional response to the antigens Env and NS1 and immune regulatory responses to NS5 and C protein. Our data support an argument that different viral products may skew the antiviral response to a more pro or anti-inflammatory outcome, with an associated impact on immunopathogenesis.

Impact of non-surgical periodontal treatment on salivary expression of cytokines related to bone metabolism.

We aimed to evaluate the impact of non-surgical periodontal treatment on the salivary expression of leptin, TNF-α, sclerostin, parathyroid hormone, osteoprotegerin, osteopontin, osteocalcin, IL-6, IL-1ß and fibroblast growth factor 23 in patients with chronic periodontitis after 1 year of follow-up. Fifteen patients with chronic periodontitis (56.0 ± SD 9.6 years) and 15 subjects with gingivitis (39.7 ± SD 4.4 years) were included in the study. Clinical periodontal parameters, such as probing pocket depth (PPD), clinical attachment level (CAL), % of plaque and bleeding on probing (BOP) were evaluated, and non-stimulated whole saliva was collected from all patients before periodontal treatment and after 1 year of follow-up. A bead-based multiplex assay measured cytokines. In the chronic periodontitis group, periodontal treatment significantly improved clinical parameters and reduced the salivary levels of IL-1ß, leptin and TNF-α (p = 0.002, 0.007 and 0.015, respectively). In the gingivitis group, there were also significant improvements in the mean patient %BOP, % Plaque, CAL and PPD. However, there were no significant changes in the cytokine's salivary levels. In conclusion, chronic periodontitis patients showed a significant reduction in the salivary levels of leptin, TNF-α and IL-1ß 1 year after periodontal treatment and a significant improvement in their clinical periodontal parameters suggesting that periodontal treatment alone can downregulate important cytokines associated with bone metabolism.

Anti-apoptotic traits in gingival tissue from patients with severe generalized chronic periodontitis.

AIM: In the present study, we aimed to evaluate the cytosolic and nuclear-mitochondrial expression of pro-apoptotic and anti-apoptotic markers in gingival tissue from patients with severe generalized chronic periodontitis (sGCP). METHODS: Twenty-four patients participated in the study: 15 (54.3 + 8.3 years) with sGCP and nine (38.2 + 5.4 years) with gingivitis alone. Gingival tissue was collected using a 1.5-mm diameter punch and homogenized using a cell disruptor. The supernatants were analyzed for the cytosolic and nuclear-mitochondrial fractions of caspase-3, Bax, Bak, Smac, lamin B, Bad, Bim, survivin, Bcl-xL, Mcl-1, and of the dimers Bcl-2/Bax, Bcl-xL/Bak and Mcl-1/Bak using a multiplex immunoassay. RESULTS: Significantly higher levels of cytosolic Bcl-xL/Bak, nuclear-mitochondrial Mcl-1/Bak, and cytosolic Bcl-xL were observed in gingival tissues from periodontitis patients compared to controls (P = 0.03, 0.03, and 0.05, respectively). The patients with gingivitis presented significantly increased levels of nuclear-mitochondrial Bad, cytosolic and nuclear-mitochondrial Bcl-2/Bax, and cytosolic Bim compared to the patients with periodontitis (P < 0.001, 0.03, 0.05, and 0.04, respectively). CONCLUSION: Significantly higher levels of anti-apoptotic markers, such as Bcl-xL/Bak, Mcl-1/Bak, and Bcl-xL, and lower levels of pro-apoptotic markers Bad and Bim in inflamed tissues indicate an anti-apoptotic trait in patients with sGCP.

Systems analysis of subjects acutely infected with the Chikungunya virus.

The largest ever recorded epidemic of the Chikungunya virus (CHIKV) broke out in 2004 and affected four continents. Acute symptomatic infections are typically associated with the onset of fever and often debilitating polyarthralgia/polyarthritis. In this study, a systems biology approach was adopted to analyze the blood transcriptomes of adults acutely infected with the CHIKV. Gene signatures that were associated with viral RNA levels and the onset of symptoms were identified. Among these genes, the putative role of the Eukaryotic Initiation Factor (eIF) family genes and apolipoprotein B mRNA editing catalytic polypeptide-like (APOBEC3A) in the CHIKV replication process were displayed. We further compared these signatures with signatures induced by the Dengue virus infection and rheumatoid arthritis. Finally, we demonstrated that the CHIKV in vitro infection of murine bone marrow-derived macrophages induced IL-1 beta production in a mechanism that is significantly dependent on the inflammasome NLRP3 activation. The observations provided valuable insights into virus-host interactions during the acute phase and can be instrumental in the investigation of new and effective therapeutic interventions.

Zika and Chikungunya Virus and Risk for Venous Thromboembolism.

A variety of viral infections are associated with hypercoagulable states and may be linked to the development of deep venous thrombosis and pulmonary embolism. The Zika and Chikungunya viral infections spread through the South and Central American continents, moving to North America in 2016, with severe cases of polyarthralgia, fever, and Guillain-Barré syndrome leading eventually to death. A decreased trend for both infections was reported in the first quarter of 2017. In this article, we report the possible association of venous thromboembolic events associated with Zika infection. After 2 cases of deep venous thrombosis in patients with acute Zika infections, D-dimer levels were measured in 172 consecutive patients who presented to the emergency department of a university hospital in an endemic region of Brazil with either Zika or Chikungunya infections confirmed by polymerase chain reaction tests. D-dimer levels were increased in 19.4% of 31 patients with Zika and in 63.8% of 141 patients with Chikungunya infections. The mechanisms behind this association are yet to be elucidated as well as the potential for venous thromboembolism prevention strategies for in-hospital patients affected by Zika and Chikungunya infections.

Systems analysis of subjects acutely infected with the Chikungunya virus

The largest ever recorded epidemic of the Chikungunya virus (CHIKV) broke out in 2004 and affected four continents. Acute symptomatic infections are typically associated with the onset of fever and often debilitating polyarthralgia/polyarthritis. In this study, a systems biology approach was adopted to analyze the blood transcriptomes of adults acutely infected with the CHIKV. Gene signatures that were associated with viral RNA levels and the onset of symptoms were identified. Among these genes, the putative role of the Eukaryotic Initiation Factor (eIF) family genes and apolipoprotein B mRNA editing catalytic polypeptide-like (APOBEC3A) in the CHIKV replication process were displayed. We further compared these signatures with signatures induced by the Dengue virus infection and rheumatoid arthritis. Finally, we demonstrated that the CHIKV in vitro infection of murine bone marrow-derived macrophages induced IL-1 beta production in a mechanism that is significantly dependent on the inflammasome NLRP3 activation. The observations provided valuable insights into virus-host interactions during the acute phase and can be instrumental in the investigation of new and effective therapeutic interventions.

Systems analysis of subjects acutely infected with the Chikungunya virus

The largest ever recorded epidemic of the Chikungunya virus (CHIKV) broke out in 2004 and affected four continents. Acute symptomatic infections are typically associated with the onset of fever and often debilitating polyarthralgia/polyarthritis. In this study, a systems biology approach was adopted to analyze the blood transcriptomes of adults acutely infected with the CHIKV. Gene signatures that were associated with viral RNA levels and the onset of symptoms were identified. Among these genes, the putative role of the Eukaryotic Initiation Factor (eIF) family genes and apolipoprotein B mRNA editing catalytic polypeptide-like (APOBEC3A) in the CHIKV replication process were displayed. We further compared these signatures with signatures induced by the Dengue virus infection and rheumatoid arthritis. Finally, we demonstrated that the CHIKV in vitro infection of murine bone marrow-derived macrophages induced IL-1 beta production in a mechanism that is significantly dependent on the inflammasome NLRP3 activation. The observations provided valuable insights into virus-host interactions during the acute phase and can be instrumental in the investigation of new and effective therapeutic interventions.

Expressão de fatores apoptóticos em biópsias de tecido gengival de pacientes com periodontite crônica severa generalizada / Expression of apoptotics factors in gingival tissue biopsies in patients with severe generalized chronic periodontitis

O objetivo deste estudo foi avaliar a expressão dos marcadores de apoptose caspase-3, survivina e dos dímeros Bcl-xL/Bak e Mcl-1/Bak, em biópsias de tecido gengival de pacientes com Periodontite Crônica Generalizada (PCG) severa. No grupo teste foram avaliados 15 pacientes portadores de PCG severa com média de idade de 54,7 + 8,4 anos. O grupo controle foi composto por 10 pacientes com gengivite com média de idade de 38,9 + 5,4 anos. Os pacientes foram avaliados em todos os elementos dentários, exceto os terceiros molares, para Profundidade de Bolsa (PB), Nível de Inserção (NI), Índice de Placa Visível (IPV) e Índice de Sangramento Gengival (ISG). Nos dentes onde as biópsias foram coletadas, também foram avaliados os critérios de Índice de Placa (IP) e Índice Gengival (IG). Foram coletadas para cada paciente, duas biópsias no grupo teste e uma no grupo controle. As biópsias foram coletadas com um punch de 1,5 mm de diâmetro, acima do fundo da bolsa ou sulco e foram armazenadas a -70° Celsius (C). Posteriormente, as amostras de tecido gengival foram pesadas, maceradas em um disruptor de células e cada biópsia foi dividida em fração citosólica e nuclear/mitocondrial. Essas frações foram analisadas em um imunoensaio com microesferas do tipo multiplex e a expressão dos marcadores de apoptose caspase-3, survivina e dos dímeros Bcl-xL/Bak e Mcl-1/Bak foi apresentada em nanograma por miligrama de tecido (ng/mg). Os dados foram apresentados em mediana e intervalo inter-quartil e avaliados pelo teste U de Mann-Whitney com p < 0,05. A correlação de Spearman foi considerada relevante sendo r > 0,6 com p < 0,01. Os valores de PB, NI e ISG foram significativamente maiores no grupo teste, comparados ao grupo controle (p < 0,001, p < 0,001, p = 0,035, respectivamente). O grupo de pacientes com periodontite apresentou, na fração nuclear/mitocondrial, concentrações de Mcl-1/Bak e survivina significativamente maiores, em comparação ao grupo de pacientes com gengivite (p = 0,05 em ambos). A correlação entre a caspase-3 e o Bcl-xL/Bak foi significantemente positiva, tanto para a fração citosólica quanto para a fração nuclear/mitocondrial (p < 0,001, p = 0,003, respectivamente). Concluímos que os pacientes com periodontite apresentaram níveis significantemente maiores dos marcadores antiapoptóticos, survivina e do dímero Mcl-1/Bak, na fração nuclear/mitocondrial, sugerindo um possível efeito de retardo no processo de apoptose.

The aim of this study was to evaluate the expression of the apoptotic markers caspase-3, survivin and the dimers Bcl-xL/Bak and Mcl-1/Bak, in gingival tissue biopsies in subjects having severe Generalized Chronic Periodontitis (GCP). In the test group were assessed 15 subjects affected by severe GCP with mean age of 54,7 + 8,4 years. The control group was formed with 10 subjects with gingivitis with mean age of 38,9 + 5,4 years. All teeth in the patients were evaluated (except the third molars) for Pocket Depth (PD), Attachment Level (AL), Visible Plaque Index (VPI) and Gingival Bleeding Index (GBI). In the teeth where the biopsies were collected, Plaque Index (PI) and Gingival Index (GI) were also measured. Two biopsies in the test group and one biopsy in the control group were collected using a 1,5 mm of diameter punch and stored at -70º Celsius (C). Just before analysis, the samples were weighted and macerated with a cell disrupter. Each biopsy was divided into cytosolic and nuclear/mitochondrial fractions. These fractions were analyzed in a multiplex microsphere immunoassay and the expression of the apoptotic markers caspase-3, survivin and the dimers Bcl-xL/Bak and Mcl-1/Bak were shown in nanogram per milligram of tissue (ng/mg). The data were presented in median and interquartile range and measured using the Mann-Whitney U Test with p < 0,05. Spearman's correlation was deemed relevant when r > 0,6 with p < 0,01. The values of PD, AL and GBI were significantly higher in the test group, when compared to the control (p < 0,001, p < 0,001, p = 0,035, respectively). The group of pacients suffering with periodontitis presented, in the nuclear/mitochondrial fraction, concentrations of Mcl-1/Bak and survivin significantly higher when compared to the control (p = 0,05 for both). The correlation between the caspase-3 and the Bcl-xL/Bak was significantly positive, from both the cytosolic and the nuclear/mitochondrial fractions (p < 0,001, p = 0,003, respectively). We conclude that the subjects having periodontitis presented significantly higher levels of the antiapoptotic markers survivin and the dimer Mcl-1/Bak, in the nuclear/mitochondrial fraction, suggesting a possible retarding effect on the process of apoptosis.

Efeito da temperatura de armazenamento na resistência coesiva de um compósito / Effect of storage temperature on the composite cohesive strength

O objetivo deste estudo foi avaliar a resistência à tração diametral de um compósito fotopolimerizável em diferentes temperaturas de armazenamento. Foram confeccionados 12 espécimes cilíndricos (n = 6),que foram divididos em dois grupos: G1- resina à temperatura ambiente (25ºC) e G2- resina utilizada imediatamente após remoção do refrigerador (5ºC). Cada espécime foi confeccionado em três incrementos, cada incremento polimerizado por 40 segundos. Realizouse o teste a uma velocidade de 1.0 mm/min até a falha. Após cada teste, a resistência à tração diametral foi calculada (MPa) e os resultados submetidos à análise estatística, pelo teste t-student (p > 0,05). Não houve efeito da temperatura de armazenamento na resistência coesiva da resina utilizada neste estudo.