ICU Admission, Invasive Mechanical Ventilation, and Mortality among Children and Adolescents Hospitalized for COVID-19 in a Private Healthcare System.

Aim: The COVID-19 pandemic devastated healthcare around the world. Data about the COVID-19 outcomes among young people are still scarce. We aim to identify factors associated with the composite outcome among children and adolescents hospitalized due to COVID-19. Methods: We performed a search in the database of a large Brazilian private healthcare system. Insured people aged 21 years or younger who were hospitalized due to COVID-19 from Feb/28th/2020 to Nov/1st/2021 were included. The primary endpoint was the composite outcome consisting of ICU admission, need for invasive mechanical ventilation, or death. Results: We evaluated 199 patients who had an index hospitalization due to COVID-19. The median monthly rate of index hospitalization was 2.7 (interquartile range [IQR], 1.6-3.9) per 100,000 clients aged 21 years or less. The median age of the patients was 4.5 years (IQR, 1.4-14.1). At the index hospitalization, the composite outcome rate was 26.6%. The composite outcome was associated with all the previous coexisting morbidities evaluated. The median follow-up was 249.0 days (IQR, 152.0-438.5). There were 27 readmissions (16 patients) within 30 days after the discharge. Conclusions: In conclusion, hospitalized children and adolescents had a composite outcome rate of 26.6% at the index hospitalization. Having previous chronic morbidity was associated with the composite.

Test of the FlashFREEZE unit in tissue samples freezing for biobanking purposes.

Availability of molecularly intact biospecimens is essential in genetic diagnostics to obtain credible results. Integrity of nucleic acids (particularly RNA) may be compromised at various steps of tissue handling, and affected by factors such as time to freeze, freezing technique and storing temperature. At the same time, freezing and storing of the biological material should be feasible and safe for the operator. Here, we compared quality of DNA and RNA from biospecimens derived from different organs (breast, colon, adrenal glands, testes, rectum and uterus) frozen either using dry ice-cooled isopentane or with FlashFREEZE unit, in order to verify if the latter is suitable for routine use in biobanking. Implementing FlashFREEZE device would enable us to limit the use of isopentane, which is potentially toxic and environmentally harmful, whilst facilitate standardization of sample freezing time. We considered factors such RNA and DNA yield and purity. Furthermore, RNA integrity and RNA/DNA performance in routine analyses, such as qPCR, next generation sequencing or microarray, were also assessed. Our results indicate that freezing of tissue samples either with FlashFREEZE unit or isopentane ensures biological material with comparable expression profiles and DNA mutation status, indicating that RNA and DNA of similar quality can be extracted from both. Therefore, our findings support the use of the FlashFREEZE device in routine use for biobanking purposes.

Potential Targeted Therapies in Ovarian Cancer.

BACKGROUND: We aimed to identify somatic pathogenic and likely pathogenic mutations using next-generation sequencing (NGS). The mutational findings were held against clinically well-described data to identify potential targeted therapies in Danish patients diagnosed with high-grade serous ovarian cancer (HGSC). METHODS: We characterized the mutational profile of 128 HGSC patients. Clinical data were obtained from the Danish Gynecological Database and tissue samples were collected through the Danish CancerBiobank. DNA was analyzed using NGS. RESULTS: 47 (37%) patients were platinum-sensitive, 32 (25%) partially platinum-sensitive, 35 (27%) platinum-resistant, and three (2%) platinum-refractory, while 11 (9%) patients did not receive chemotherapy. Overall, 27 (21%) had known druggable targets. Twelve (26%) platinum-sensitive patients had druggable targets for PARP inhibitors: one for tyrosine kinase inhibitors and one for immunotherapy treatment. Eight (25%) partially platinum-sensitive patients had druggable targets: seven were eligible for PARP inhibitors and one was potentially eligible for alpesilib and hormone therapy. Seven (20%) platinum-resistant patients had druggable targets: six (86%) were potentially eligible for PARP inhibitors, one for immunotherapy, and one for erdafitinib. CONCLUSIONS: PARP inhibitors are the most frequent potential targeted therapy in HGSC. However, other targeted therapies remain relevant for investigation according to our mutational findings.

Silver chitosan nanocomposites as a potential treatment for superficial candidiasis.

Silver compounds are widely known for their antimicrobial activity, but can exert toxic effects to the host. Among the strategies to reduce its toxicity, incorporation into biopolymers has shown promising results. We investigated the green syntheses of silver nanoparticles (AgNPs) and their functionalization in a chitosan matrix (AgNPs@Chi) as a potential treatment against Candida spp. Inhibitory concentrations ranging between 0.06 and  1 µg/ml were observed against distinct Candida species. Nanocomposite-treated cells displayed cytoplasmic degeneration and a cell membrane and wall disruption. Silver nanocomposites in combination with fluconazole and amphotericin B showed an additive effect when analyzed by the Bliss method. The low cytotoxicity displayed in mammalian cells and in the Galleria mellonella larvae suggested their potential use in vivo. When tested as a topical treatment against murine cutaneous candidiasis, silver nanocomposites reduced the skin fungal burden in a dose-response behavior and favored tissue repair. In addition, the anti-biofilm effect of AgNPs@Chi in human nail model was demonstrated, suggesting that the polymeric formulation of AgNPs does not affect antifungal activity even against sessile cells. Our results suggest that AgNPs@Chi seems to be a less toxic and effective topical treatment for superficial candidiasis. LAY SUMMARY: This study demonstrated the efficacy of silver nanoparticles (AgNPs) in inhibiting the growth of Candida. AgNPs incorporated in chitosan displayed a reduced toxicity. Tests in infected mice showed the effectiveness of the treatment. AgNPs-chitosan could be an alternative to combat candidiasis.

Functionalized microchannels as xylem-mimicking environment: Quantifying X. fastidiosa cell adhesion.

Microchannels can be used to simulate xylem vessels and investigate phytopathogen colonization under controlled conditions. In this work, we explore surface functionalization strategies for polydimethylsiloxane and glass microchannels to study microenvironment colonization by Xylella fastidiosa subsp. pauca cells. We closely monitored cell initial adhesion, growth, and motility inside microfluidic channels as a function of chemical environments that mimic those found in xylem vessels. Carboxymethylcellulose (CMC), a synthetic cellulose, and an adhesin that is overexpressed during early stages of X. fastidiosa biofilm formation, XadA1 protein, were immobilized on the device's internal surfaces. This latter protocol increased bacterial density as compared with CMC. We quantitatively evaluated the different X. fastidiosa attachment affinities to each type of microchannel surface using a mathematical model and experimental observations acquired under constant flow of culture medium. We thus estimate that bacterial cells present ∼4 and 82% better adhesion rates in CMC- and XadA1-functionalized channels, respectively. Furthermore, variable flow experiments show that bacterial adhesion forces against shear stresses approximately doubled in value for the XadA1-functionalized microchannel as compared with the polydimethylsiloxane and glass pristine channels. These results show the viability of functionalized microchannels to mimic xylem vessels and corroborate the important role of chemical environments, and particularly XadA1 adhesin, for early stages of X. fastidiosa biofilm formation, as well as adhesivity modulation along the pathogen life cycle.

Slave Voyages: Reflections on Data Sculptures.

This pictorial presents the development of a data sculpture, followed by our reflections inspired by Research through Design (RtD) and Dahlstedt's process-based model of artistic creativity. We use the notion of negotiation between concept and material representation to reflect on the ideation, design process, production, and the exhibition of "Slave Voyages" - a set of data sculptures that depicts slave traffic from Africa to the American continent. The work was initially produced as an assignment on physicalization for the Design course at the Federal University of Rio de Janeiro. Our aim is to open discussion on material representation and negotiation in the creative process of data physicalization.

Frequency and coexistence of KRAS, NRAS, BRAF and PIK3CA mutations and occurrence of MMR deficiency in Danish colorectal cancer patients.

The MAPK signalling genes KRAS, NRAS and BRAF and the PIK3CA gene are routinely investigated for mutations in the diagnostic routine of colorectal cancer. Few studies have reported co-existing mutations in these genes with clinical relevance, while some have been previously regarded as mutually exclusive. We set to investigate the frequency and co-occurrent mutations in these targets, and the occurrence of mismatch repair deficiency (dMMR) in a large cohort of Danish colorectal cancers. 1000 colorectal tumours were sequenced as part of our diagnostic workflow for KRAS, NRAS, BRAF and PIK3CA mutations using next-generation sequencing (NGS) and analysed by immunohistochemistry (IHC) for loss of the MMR proteins, MLH1, PMS2, MSH2 and MSH6. Co-existing mutations in 12 patients (1.2%) occurred as multiple mutations in the same gene or spread across several genes (KRAS, NRAS and/or BRAF). The frequency of single mutations in the genes occurred with a frequency similar to previously reported, except for a higher frequency of BRAF mutations (18.0%). We found dMMR in 14.6% of the cases with a majority lacking expression of both MLH1 and PMS2. BRAF mutations were only present in dMMR cases involving MLH1 and/or PMS2. Our findings suggest that co-existing mutations occur, except for the hotspot BRAF V600E, which is mutually exclusive with KRAS/NRAS mutations. Therefore, instead of single gene alterations from the MAPK signalling, assessing co-occurrence of mutations within one or more of those genes should also be accounted. This may impact future oncological treatments and should be considered in the diagnostic workflow.

Nanoparticles Loaded with a New Thiourea Derivative: Development and In vitro Evaluation Against Leishmania amazonensis.

BACKGROUND: Leishmaniasis is a neglected tropical disease caused by protozoa of the genus Leishmania. Current treatments are restricted to a small number of drugs that display both severe side effects and a potential for parasites to develop resistance. A new N-(3,4-methylenedioxyphenyl)-N'- (2-phenethyl) thiourea compound (thiourea 1) has shown promising in vitro activity against Leishmania amazonensis with an IC50 of 54.14 µM for promastigotes and an IC50 of 70 µM for amastigotes. OBJECTIVE: To develop a formulation of thiourea 1 as an oral treatment for leishmaniasis, it was incorporated into Nanoparticles (NPs), a proven approach to provide long-acting drug delivery systems. METHODS: Poly (D,L-Lactic-co-Glycolic Acid) (PLGA) polymeric NPs containing thiourea 1 were obtained through a nanoprecipitation methodology associated with solvent evaporation. The NPs containing thiourea 1 were characterized for Encapsulation Efficiency (EE%), reaction yield (% w/w), surface charge, particle size and morphology by Transmission Electron Microscopy (TEM). RESULTS: NPs with thiourea 1 showed an improved in vitro leishmanicidal activity with a reduction in its cytotoxicity against macrophages (CC50>100 µg/mL) while preserving its IC50 against intracellular amastigotes (1.46 ± 0.09 µg/mL). This represents a parasite Selectivity Index (SI) of 68.49, which is a marked advancement from the reference drug pentamidine (SI = 30.14). CONCLUSION: The results suggest that the incorporation into NPs potentiated the therapeutic effect of thiourea 1, most likely by improving the selective delivery of the drug to the phagocytic cells that are targeted for infection by L. amazonensis. This work reinforces the importance of nanotechnology in the acquisition of new therapeutic alternatives for oral treatments.

Evaluation of the bioaccumulation kinetics of toxic metals in fish (A. brasiliensis) and its application on monitoring of coastal ecosystems.

This study proposes a pro-active approach for evaluations of methylmercury (MeHg), total mercury (THg), arsenic (As), cadmium (Cd) and lead (Pb) in situ bioaccumulation in fish (Atherinella brasiliensis) muscles, using specimens from the external sector of Guanabara Bay as a study case. This approach included an hierarchical sequence: analysis of the pollutants concentrations and their comparison to safety criteria; correlations between specimens concentrations vs length (as a proxy of exposure time); projections of concentrations in key lengths (sexual maturation, asymptotic, length limits for fishing and median of fish population) through polynomial regressions, dose-response analysis (Probit), decreasing curves and incorporation rates (using only three length intervals). The incorporation rates were ascending for MeHg and THg (continued bioaccumulation) and descending for As, Pb and Cd (possible biological dilution). The projections were satisfactory, evidencing their use for an improvement on the risks monitoring of fishing and fish consumption by humans in coastal environments.

Maxillary stability following Le Fort I osteotomy using prebent plates and wire fixation in patients undergoing surgery for OSAS.

The purpose of this study was to analyze the stability of Le Fort I maxillary advancement in the vertical and horizontal directions using a combination of wire and rigid fixation in patients undergoing surgery to treat obstructive sleep apnea (OSA). Wire osteosynthesis can be performed quicker and at a reduced cost. The lateral cephalograms of 21 patients were evaluated preoperatively (T0), immediately postoperatively (T1), and at least 6 months postoperatively (T2). Four cephalometric points were used to measure movement in the horizontal and vertical directions. Mean values were determined, and data were statistically analyzed by ANOVA to determine differences between time points. Of the four points analyzed, the average maxillary advancement in the horizontal direction was 7.48 mm and the relapse was 0.56 mm with absence of statistically significant differences between the measurements taken (T1) and (T2). There was a 5% probability of error in the vertical movements at the points I and posterior nasal spine. The combination of two pre-bent plates in piriform aperture with osteosynthesis using surgical steel wires in the zygomatic buttress in patients undergoing maxillary surgery for OSA stabilized the large horizontal maxillary advancements and enhanced vertical stability in the first molar and A point regions.

Custom-Made Titanium Miniplates Associated With Ultrahigh-Molecular-Weight Polyethylene Graft in Orthognathic Surgery: An Adjunct to Maxillary Advancement.

Patients with considerable maxillomandibular anteroposterior discrepancies and maxillary hypoplasia require corrective treatment through orthognathic surgery. However, in the treatment of severe maxillary retrognathism, it is necessary to reconstruct areas of bone deficiency through grafting techniques in addition to maxillary advancement using only the Le Fort I osteotomy. Treatment in these patients is more challenging and requires high surgical predictability. Alloplastic materials often have been used for the reconstruction of poor bone contours. Ultrahigh-molecular-weight polyethylene (UHMWPE) is currently an excellent filler material for poor bone regions and is a good substitute for autografts and other alloplastic materials for its unique properties, including high biocompatibility. Insertion of this material in the fixation system customized for virtually planned orthognathic surgeries is an innovative technique. This report describes the insertion of UHMWPE into custom-made titanium miniplates manufactured by computer-aided design and computer-aided manufacturing technology for orthognathic surgery consisting of maxillary advancement and mandibular retrusion to treat a patient with Crouzon syndrome, Class III malocclusion, and severe maxillary retrognathism.

Citrulline Malate Does Not Improve Muscle Recovery after Resistance Exercise in Untrained Young Adult Men.

The effects of citrulline malate (CM) on muscle recovery from resistance exercise remains unknown. We aimed to determine if citrulline malate supplementation improves muscle recovery after a single session of high-intensity resistance exercise (RE) in untrained young adult men. Nine young adult men (24.0 ± 3.3 years) participated in a double-blind crossover study in which they received 6 g of CM and placebo (PL) on two occasions, separated by a seven-day washout period. Each occasion consisted of a single session of high-intensity RE (0 h) and three subsequent fatigue tests sessions (at 24, 48, and 72 h) to assess the time course of muscle recovery. During the tests sessions, we assessed the following variables: number of maximum repetitions, electromyographic signal (i.e., root mean square (RMS) and median frequency (MF)), muscle soreness and perceived exertion, as well as blood levels of creatine kinase (CK), lactate, insulin, and testosterone:cortisol ratio. CK levels increased at 24 h post-exercise and remained elevate at 48 and 72 h, with no difference between CM and PL conditions. Muscle soreness increased at 24 h post-exercise, which progressively returned to baseline at 72 h in both conditions. Lactate levels increased immediately post-exercise and remained elevated at 24, 48, and 72 h in both conditions. No significant treatment × time interaction was found for all dependents variables (maximum repetitions, perceived exertion, CK, lactate, RMS, MF, and testosterone:cortisol ratio) during the recovery period. In conclusion, our data indicate that CM supplementation (single 6 g dose pre-workout) does not improve the muscle recovery process following a high-intensity RE session in untrained young adult men.

Efficacy of red and infrared lasers in treatment of temporomandibular disorders--a double-blind, randomized, parallel clinical trial.

AIM: Low-level laser therapy has still not been well established, and it is important to define a standardized protocol for the treatment of temporomandibular disorders (TMDs) using low level laser. There is no consensus on controlled clinical trials concerning the best option for laser therapy with regard to wavelength. The aim of this study was to evaluate the efficacy of red and infrared laser therapy in patients with TMD, using a randomized parallel-group double-blind trial. METHODOLOGY: Each hemiface of 19 subjects was randomized to receive intervention, in a total of 116 sensitive points. Pain was measured at baseline and time intervals of 24 hours, 30 days, 90 days, and 180 days after treatment. Irradiation of 4 J/cm2 in the temporomandibular joints and 8 J/cm(2) in the muscles was used in three sessions. RESULTS: Both treatments had statistically significant results (P<0.001); there was statistical difference between them at 180 days in favor of the infrared laser (P=0.039). There was improvement in 24 hours, which extended up to 180 days in both groups. CONCLUSION: Both lasers are effective in the treatment and remission of TMD symptoms.