Colchicine reduces the activation of NLRP3 inflammasome in COVID-19 patients.

OBJECTIVE: To evaluate whether colchicine treatment was associated with the inhibition of NLRP3 inflammasome activation in patients with COVID-19. METHODS: We present a post hoc analysis from a double-blinded placebo-controlled randomized clinical trial (RCT) on the effect of colchicine for the treatment of COVID-19. Serum levels of NOD-like receptor protein 3 (NLRP3) inflammasome products-active caspase-1 (Casp1p20), IL-1ß, and IL-18-were assessed at enrollment and after 48-72 h of treatment in patients receiving standard-of-care (SOC) plus placebo vs. those receiving SOC plus colchicine. The colchicine regimen was 0.5 mg tid for 5 days, followed by 0.5 mg bid for another 5 days. RESULTS: Thirty-six patients received SOC plus colchicine, and thirty-six received SOC plus placebo. Colchicine reduced the need for supplemental oxygen and the length of hospitalization. On Days 2-3, colchicine lowered the serum levels of Casp1p20 and IL-18, but not IL-1ß. CONCLUSION: Treatment with colchicine inhibited the activation of the NLRP3 inflammasome, an event triggering the 'cytokine storm' in COVID-19. TRIAL REGISTRATION NUMBERS: RBR-8jyhxh.

In vitro characterization of Trichophyton rubrum and T. mentagrophytes biofilms.

Dermatophytes are fungi responsible for a disease known as dermatophytosis. Biofilms are sessile microbial communities surrounded by extracellular polymeric substances (EPS) with increased resistance to antimicrobial agents and host defenses. This paper describes, for the first time, the characteristics of Trichophyton rubrum and T. mentagrophytes biofilms. Biofilm formation was analyzed by light microscopy, scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM) as well as by staining with crystal violet and safranin. Metabolic activity was determined using the XTT reduction assay. Both species were able to form mature biofilms in 72 h. T. rubrum biofilm produced more biomass and EPS and was denser than T. mentagrophytes biofilm. The SEM results demonstrated a coordinated network of hyphae in all directions, embedded within EPS in some areas. Research and characterization of biofilms formed by dermatophytes may contribute to the search of new drugs for the treatment of these mycoses and might inform future revisions with respect to the dose and duration of treatment of currently available antifungals.

Comparison of Blue and Infrared Light Transmission Through Dental Tissues and Restorative Materials.

OBJECTIVES: The depth of cure using blue-light photocuring units (BL) is limited by tooth structure and qualities of the restorative material through which the activating wavelength must pass. Recent developments incorporate an infrared (IR) activated upconversion (UC) fluorescence of a lining agent filled with nanocrystals of NaYF4 and doped with YB+3 and Tm+3 that emit both blue and violet light locally at the interface of the liner and restorative resin. The purpose of this study was to evaluate the BL and 975 nm infrared (IR) light power transmission through dental tissues and restorative materials. METHODS AND MATERIALS: Power transmissions of the IR laser (975 nm) and a monowave blue-only light-curing unit (Bluephase 16i) through dental tissues (enamel, dentin, and enamel/dentin junction, or DEJ), eight (8) various dental resin composites, and eight (8) dental ceramics, each at four thicknesses (1, 2, 3 and 4 mm) were evaluated (n=5) using a thermopile sensor (PM10, Coherent Inc) connected to a laser power meter (Fieldmate, Coherent Inc). Power transmission values of each light source and restorative material were subjected to analysis of variance and Tukey test at a pre-set alpha of 0.05. RESULTS: A linear correlation (r=0.9884) between the supplied current and emitted IR power of the laser diode was found, showing no statistical power reduction with increased distances (collimated beam). For tooth tissues, the highest power transmissions for both light sources were observed using 1.0 mm enamel while the lowest values were found for 2.0 mm dentin and an association of 2.0 mm DEJ and 1.0 mm dentin. The only group where IR demonstrated significantly higher transmission when compared to BL was 1.0 mm enamel. For all resin composites and dental ceramics, increased thickness resulted in a reduction of IR power transmission (except for EverX Posterior fiber-reinforced composite and e.max HT ceramic). IR resulted in higher transmission through all resin composites, except for Tetric EvoCeram White. The highest BL transmission was observed for SDR Flow, at all thicknesses. Higher IR/BL ratios were observed for EverX Posterior, Herculite Ultra, and Lava Ultimate, while the lowest ratio was observed for Tetric EvoCeram White. Reduced translucency shades within the same material resulted in lower power ratio values, especially for BL transmission. Higher IR/BL ratios were observed for e.Max LT, VitaVM7 Base Dentin, and e.max CAD HT, while the lowest values were found for VitaVM7 Enamel and Paradigm C. CONCLUSION: IR power transmission through enamel was higher when compared to blue light, while no difference was observed for dentin. The power transmission of IR was higher than BL for resin composites, except for a high value and low chroma shade. Fiber-reinforced resin composite demonstrated the highest IR/BL power transmission ratio. A greater IR/BL ratio was observed for lower translucency ceramics when compared to high translucency.

Microplate alamarBlue assay for Paracoccidioides susceptibility testing.

CLSI method M27-A3 is not available for use with dimorphic fungi, such as those of the Paracoccidioides genus. In this study, we developed a microdilution method and added the alamarBlue reagent to test the responses of Paracoccidioides brasiliensis and Paracoccidioides lutzii against amphotericin B and itraconazole antifungals. The test proved to be sensitive, practical, and inexpensive and can be used to monitor the activity of low-growth microorganisms and their response to various drugs.

Cryptococcosis: epidemiology, fungal resistance, and new alternatives for treatment.

Cryptococcosis is an important systemic mycosis and the third most prevalent disease in human immunodeficiency virus (HIV)-positive individuals. The incidence of cryptococcosis is high among the 25 million people with HIV/acquired immunodeficiency syndrome (AIDS), with recent estimates indicating that there are one million cases of cryptococcal meningitis globally per year in AIDS patients. In Cryptococcus neoformans, resistance to azoles may be associated with alterations in the target enzyme encoded by the gene ERG11, lanosterol 14α-demethylase. These alterations are obtained through mutations, or by overexpressing the gene encoding. In addition, C. gattii and C. neoformans present a heteroresistance phenotype, which may be related to increased virulence. Other species beyond C. neoformans and C. gattii, such as C. laurentii, have been diagnosed mainly in patients with immunosuppression. Infections of C. albidus have been isolated in cats and marine mammals. Recent evidence suggests that the majority of infections produced by this pathogen are associated with biofilm growth, which is also related with increased resistance to antifungal agents. Therefore, there is a great need to search for alternative antifungal agents for these fungi. The search for new molecules is currently occurring from nanoparticle drugs of plant peptide origin. This article presents a brief review of the literature regarding the epidemiology of cryptococcosis, as well as fungal resistance and new alternatives for treatment.

Effect of High-radiant Emittance and Short Curing Time on Polymerization Shrinkage Vectors of Bulk Fill Composites.

PURPOSE: To evaluate the effect of short curing time using a high-radiant emittance light on polymerization shrinkage vectors in different consistency bulk-fill composites (BFRCs) using micro-computed tomography. METHODS AND MATERIALS: Radiopaque zirconia fillers were homogeneously incorporated and functioned as radiopaque tracers into two regular-paste: TBFill (Tetric EvoCeram Bulk Fill) and TPFill (Tetric PowerFill), and two flowable (n=6): TBFlow (Tetric EvoFlow Bulk Fill) and TPFlow (Tetric PowerFlow) resin composites. Class I cavities (4 mm depth × 4 mm length × 4 mm width) were 3D-printed and filled in a single increment: TBFill and TBFlow were light-activated using a Bluephase Style 20i (10 seconds in high-mode); TPFill and TPFlow were light-activated using a Bluephase PowerCure (three seconds). The same adhesive system (Adhese Universal) was used for all groups. Microcomputed tomography scans were obtained before and after light-activation. Filler particle movement was identified by polymerization shrinkage vectors at five depths (from 0-4 mm): top, top-middle, middle, middle-bottom and bottom. RESULTS: TPFlow showed the lowest total vector displacement, followed by TBFlow, TBFill and TPFill, significantly different among each other (p<0.05). Generally, BFRCs showed decreased vector displacement with increased depth, and higher displacement at the top-surface (p<0.05). Qualitative analysis showed a similar pattern of vector magnitude and displacement for groups TBFill and TPFill, with displacement vectors on occlusal (top) surfaces toward the center of the restoration from the top to middle areas, and relatively limited displacement at the bottom. TBFlow and TPFlow showed more displacement on the occlusal (top). CONCLUSIONS: Short curing time with high-radiant emittance on fast-curing BFRCs was shown to be a feasible option in terms of vector displacement. Flowable BFRCs presented lower vector displacement than their regular-viscosity versions.

Effect of Ceramic Conditioners on Surface Morphology, Roughness, Contact Angle, Adhesion, Microstructure, and Composition of CAD/CAM Ceramics.

The objective of this study was to investigate the effect of surface treatments in maximum profile valley depth (Rv), surface roughness (Sa), contact angle (Ca), shear bond strength (SBS) of a light-cured resin cement, microstructure, and composition of two CAD/CAM ceramics: Cerec Blocs/Dentsply Sirona (feldspathic - FEL) and Empress CAD/Ivoclar Vivadent (leucite-reinforced - LEU). The ceramic specimens were submitted to six surface treatments: (1) 5% hydrofluoric acid (HF) - 20 seconds; (2) 5% HF - 60 seconds; (3) 10% HF - 20 seconds; (4) 10% HF - 60 seconds; (5) self-etching ceramic primer (MEP) - 20 seconds; and (6) MEP - 60 seconds. Specimens treated with HF received silane application for 1 minute. Rv and Sa were evaluated in a confocal laser microscope (n=10) and the Ca in a goniometer (n=6). For the SBS test, two resin cement cylinders were bonded to each specimen, one tested after 24 hours in distilled water storage (37°C) and the other after one year (n=10). The microstructures and compositions were analyzed by scanning electron microscopy (SEM) and energy dispersive x-ray spectroscopy (EDS) (n=4). Rv, Sa, and Ca data were analyzed by two-way ANOVA, and the SBS data by three-way ANOVA, all followed by post-hoc Tukey's test (α=0.05). Results: Surface treatments with HF always produced higher Rv and Sa, and lower Ca than MEP 60s for both ceramics. After 1-year water storage, FEL ceramic treated with MEP for 60 seconds produced higher SBS values than HF treatments, whereas for LEU ceramic there were no differences among treatments. Application of MEP for 20 or 60 seconds produced no difference in Rv, Sa, Ca, and SBS for both ceramics. Surface treatments produced considerable differences in ceramic topography, but subtle ones in composition, for both ceramics. Conclusions: The results indicated that for FEL ceramic, MEP application for 60 seconds yielded higher SBS compared with HF etching, while for LEU ceramic the surface treatments did not influence the SBS results.

No association between chronic cerebrospinal venous insufficiency and pediatric-onset multiple sclerosis.

OBJECTIVE: Chronic cerebrospinal venous insufficiency (CCSVI) was hypothesized to play a causative role in multiple sclerosis (MS). The assessment of pediatric-onset MS (POMS) may provide a unique window of opportunity to study hypothesized risk factors in close temporal association with the onset of the disease. METHODS: Internal jugular veins, vertebral veins and intracranial veins were evaluated with extracranial and intracranial ultrasound in 15 POMS and 16 healthy controls. Assessor's blinding was maintained during the study. We considered subjects positive to CCSVI when at least two criteria were fulfilled. RESULTS: CCSVI frequency was comparable between POMS and controls (p > 0.05). Clinical features were not significantly different between CCSVI-positive and CCSVI-negative patients. CONCLUSIONS: Our findings add to previous data pointing against a causative role of CCSVI in MS.

Adhesion of Histoplasma capsulatum to pneumocytes and biofilm formation on an abiotic surface.

The pathogenic fungus, Histoplasma capsulatum, causes the respiratory and systemic disease 'histoplasmosis'. This disease is primarily acquired via inhalation of aerosolized microconidia or hyphal fragments of H. capsulatum. Evolution of this respiratory disease depends on the ability of H. capsulatum yeasts to survive and replicate within alveolar macrophages. It is known that adhesion to host cells is the first step in colonization and biofilm formation. Some microorganisms become attached to biological and non-biological surfaces due to the formation of biofilms. Based on the importance of biofilms and their persistence on host tissues and cell surfaces, the present study was designed to investigate biofilm formation by H. capsulatum yeasts, as well as their ability to adhere to pneumocyte cells. H. capsulatum biofilm assays were performed in vitro using two different clinical strains of the fungus and biofilms were characterized using scanning electron microscopy. The biofilms were measured using a 2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-5-[(phenylamino)carbonyl]-2H-tetrazolium-hydroxide (XTT) reduction assay. The results showed that both the H. capsulatum strains tested were very efficient at adhering to host cells and forming biofilm. Therefore, this is a possible survival strategy adopted by this fungus.