Multicenter study of the natural history and therapeutic responses of patients with chikungunya, focusing on acute and chronic musculoskeletal manifestations - a study protocol from the clinical and applied research in Chikungunya (REPLICK network).

BACKGROUND: Chikungunya is associated with high morbidity and the natural history of symptomatic infection has been divided into three phases (acute, post-acute, and chronic) according to the duration of musculoskeletal symptoms. Although this classification has been designed to help guide therapeutic decisions, it does not encompass the complexity of the clinical expression of the disease and does not assist in the evaluation of the prognosis of severity nor chronic disease. Thus, the current challenge is to identify and diagnose musculoskeletal disorders and to provide the optimal treatment in order to prevent perpetuation or progression to a potentially destructive disease course. METHODS: The study is the first product of the Clinical and Applied Research Network in Chikungunya (REPLICK). This is a prospective, outpatient department-based, multicenter cohort study in Brazil. Four work packages were defined: i. Clinical research; ii) Translational Science - comprising immunology and virology streams; iii) Epidemiology and Economics; iv) Therapeutic Response and clinical trials design. Scheduled appointments on days 21 (D21) ± 7 after enrollment, D90 ± 15, D120 ± 30, D180 ± 30; D360 ± 30; D720 ± 60, and D1080 ± 60 days. On these visits a panel of blood tests are collected in addition to the clinical report forms to obtain data on socio-demographic, medical history, physical examination and questionnaires devoted to the evaluation of musculoskeletal manifestations and overall health are performed. Participants are asked to consent for their specimens to be maintained in a biobank. Aliquots of blood, serum, saliva, PAXgene, and when clinically indicated to be examined, synovial fluid, are stored at -80° C. The study protocol was submitted and approved to the National IRB and local IRB at each study site. DISCUSSION: Standardized and harmonized patient cohorts are needed to provide better estimates of chronic arthralgia development, the clinical spectra of acute and chronic disease and investigation of associated risk factors. This study is the largest evaluation of the long-term sequelae of individuals infected with CHIKV in the Brazilian population focusing on musculoskeletal manifestations, mental health, quality of life, and chronic pain. This information will both define disease burden and costs associated with CHIKV infection, and better inform therapeutic guidelines.

Serum biomarkers associated with SARS-CoV-2 severity.

Immunity with SARS-CoV-2 infection during the acute phase is not sufficiently well understood to differentiate mild from severe cases and identify prognostic markers. We evaluated the immune response profile using a total of 71 biomarkers in sera from patients with SARS-CoV-2 infection, confirmed by RT-PCR and controls. We correlated biological marker levels with negative control (C) asymptomatic (A), nonhospitalized (mild cases-M), and hospitalized (severe cases-S) groups. Among angiogenesis markers, we identified biomarkers that were more frequently elevated in severe cases when compared to the other groups (C, A, and M). Among cardiovascular diseases, there were biomarkers with differences between the groups, with D-dimer, GDF-15, and sICAM-1 higher in the S group. The levels of the biomarkers Myoglobin and P-Selectin were lower among patients in group M compared to those in groups S and A. Important differences in cytokines and chemokines according to the clinical course were identified. Severe cases presented altered levels when compared to group C. This study helps to characterize biological markers related to angiogenesis, growth factors, heart disease, and cytokine/chemokine production in individuals infected with SARS-CoV-2, offering prognostic signatures and a basis for understanding the biological factors in disease severity.

Influence of the self-adhering strategy on microhardness, sorption, solubility, color stability, and cytotoxicity compared to bulk-fill and conventional resin composites.

OBJECTIVES: To analyze and compare, in vitro, the microhardness, sorption, solubility, color stability, and cytotoxicity of three types of resin composites: self-adhesive (SARC) (Dyad Flow (DF)/Kerr), bulk-fill (Filtek Bulk Fill Flow (FBF)/3 M ESPE), and conventional (Filtek Z350XT Flow (Z350)/3 M ESPE). MATERIALS AND METHODS: Thirty cylindrical specimens were prepared using a split metal mold (15 mm × 1 mm), divided into 3 groups (n = 10) according to the material used. Vickers hardness (VH) was calculated from three indentations (300gf/15 s) per specimen. The sorption and solubility were measured according to the ISO 4049:2009 specification after storing in distilled water for 7 days. The color of each resin composite was measured using a portable digital spectrophotometer according to the CIELAB system. After a 7-day immersion in coffee, the color variation (∆E) was calculated. Following the ISO 10993:2012, the cytotoxicity in Vero cells was evaluated through the MTT assay. The results were analyzed using the Kruskal-Wallis test to compare the studied groups. The Wilcoxon test was used to compare the assessments in each studied group. For cytotoxicity analysis, the data were compared by the ANOVA test (α = 0.05). RESULTS: DF showed the lowest VH (28.67), highest sorption (0.543 µg/mm3) and solubility (1.700 µg/mm3), and higher ∆E after 7 days of coffee immersion (p = 0.008). The resin composites studied were considered non-cytotoxic. CONCLUSIONS: The SARC presented inferior mechanical and physical-chemical properties than bulk-fill and conventional resin composites, with comparable cytotoxicity against Vero cells. CLINICAL RELEVANCE: The simplification of the clinical protocol of SARC can minimize the number of possible failures during the restorative technique. However, considering their inferior physical and mechanical properties, their coverage with materials of higher mechanical properties and physical-chemical stability should be considered.

Overexpression of UDP-Glucose 4-Epimerase Is Associated with Differentiation Grade of Gastric Cancer.

The UDP-glucose 4-epimerase (GALE) is a glycosyltransferase, which acts on protein and lipid glycosylation in normal and neoplastic cells. This study is aimed at investigating the differential tissue expression of GALE and its possible association with clinical-pathological parameters and the outcome of gastric adenocarcinoma patients. Seventy-one patients were evaluated in relation to GALE expression by immunohistochemistry. Our results showed that 48 (67.6%) patients were GALE positive and 23 (32.4%) negative. Positive staining was present on well-differentiated and moderate-differentiated histological grade of gastric adenocarcinomas (p < 0.0001). There was no significant association with outcome parameters (p > 0.05). Besides that, our results corroborated with the validation cohort analysis, where the expression of GALE mRNA was also associated with the histological grade (p < 0.001). These results suggest a possible use of this enzyme as a biomarker for well and moderately differentiated tumors.