Bleaching efficacy of in-office dental bleaching with different application protocols: a single-blind randomized controlled trial.

OBJECTIVE: This randomized controlled trial aimed to evaluate the equivalence in the color change, adverse effects, self-perception (AS) and the impact on oral condition (IO) of participants submitted to different application protocols of in-office dental bleaching. MATERIALS AND METHODS: 165 participants were bleached with a 35% hydrogen peroxide gel (Total Blanc Office One-Step, DFL), according to the following protocols: (1) 2 applications of 20-min each (2 × 20 min); (2) 1 × 40-min and; (3) 1 × 30-min. The color change was evaluated with the Vita Easyshade spectrophotometer, Vita Classical and Vita Bleachedguide scales. The intensity and risk of tooth sensitivity (TS) and gingival irritation (GI) were recorded using a 0-10 visual analogue scale (VAS). AS and IO was assessed before and after the bleaching procedure using the Orofacial Aesthetic Scale and Oral Health Impact Profile-14, respectively. RESULTS: Equivalent color change were observed (p < 0.001), with no significant difference between groups. The group 2 × 20 min presented the highest risk of TS (76%, 95% CI 63 to 85), compared to the 1 × 30 min (p < 0.04). The intensity of TS and GI and the risk of GI was similar between groups (p > 0.31). Irrespectively of the group (p = 0.32), significant improvements were observed for all items of AS and IO after bleaching (p < 0.02). CONCLUSIONS: The 1 × 30 min protocol produced equivalent color change to the other bleaching protocols with reduced risk of TS and shorter application time. CLINICAL RELEVANCE: A more simplified application regimen of a single application of 30 min yields effective bleaching and patient satisfaction while minimizing undesirable side effects and improving patient satisfaction.

Single-cell adhesive profiling in an optofluidic device elucidates CD8+ T lymphocyte phenotypes in inflamed vasculature-like microenvironments.

Tissue infiltration by circulating leukocytes occurs via adhesive interactions with the local vasculature, but how the adhesive quality of circulating cells guides the homing of specific phenotypes to different vascular microenvironments remains undefined. We developed an optofluidic system enabling fluorescent labeling of photoactivatable cells based on their adhesive rolling velocity in an inflamed vasculature-mimicking microfluidic device under physiological fluid flow. In so doing, single-cell level multidimensional profiling of cellular characteristics could be characterized and related to the associated adhesive phenotype. When applied to CD8+ T cells, ligand/receptor expression profiles and subtypes associated with adhesion were revealed, providing insight into inflamed tissue infiltration capabilities of specific CD8+ T lymphocyte subsets and how local vascular microenvironmental features may regulate the quality of cellular infiltration. This methodology facilitates rapid screening of cell populations for enhanced homing capabilities under defined biochemical and biophysical microenvironments, relevant to leukocyte homing modulation in multiple pathologies.

Microengineered In Vitro Assays for Screening and Sorting Manufactured Therapeutic T Cells.

Adoptively transferred T cells constitute a major class of current and emergent cellular immunotherapies for the treatment of disease, including but not limited to cancer. Although key advancements in molecular recognition, genetic engineering, and manufacturing have dramatically enhanced their translational potential, therapeutic potency remains limited by poor homing and infiltration of transferred cells within target host tissues. In vitro microengineered homing assays with precise control over micromechanical and biological cues can address these shortcomings by enabling interrogation, screening, sorting, and optimization of therapeutic T cells based on their homing capacity. In this article, the working principles, application, and integration of microengineered homing assays for the mechanistic study of biophysical and biomolecular cues relevant to homing of therapeutic T cells are reviewed. The potential for these platforms to enable scalable enrichment and screening of next-generation manufactured T cell therapies for cancer is also discussed.

Veno-venous extracorporeal membrane oxygenation in patients with SARS-CoV-2 pneumonia in Brazil: a case series.

OBJECTIVE: The world has been suffering from the COVID-19 pandemic. Some COVID-19 patients develop severe viral pneumonia, requiring mechanical ventilation and measures to treat refractory hypoxemia, such as a protective ventilation strategy, prone positioning, and the use of veno-venous extracorporeal membrane oxygenation (VV-ECMO). We describe a case series of 30 COVID-19 patients who needed VV-ECMO at the Hospital Alemão Oswaldo Cruz, located in the city of São Paulo, Brazil. METHODS: We included all patients who required VV-ECMO due to COVID-19 pneumonia between March of 2020 and June of 2021. RESULTS: Prior to VV-ECMO, patients presented with the following median scores: SOFA score, 11; APPS score, 7; Respiratory ECMO Survival Prediction score, 2; and Murray score, 3.3. The 60-day-in-hospital mortality was 33.3% (n = 10). CONCLUSIONS: Although our patients had a highly severe profile, our results were similar to those of other cohort studies in the literature. This demonstrates that VV-ECMO can be a good tool even in a pandemic situation when it is managed in an experienced center.

Assessment of the Interferon-Lambda-3 Polymorphism in the Antibody Response to COVID-19 in Older Adults Seropositive for CMV.

BACKGROUND: Here, we investigated the impact of IFN-lambda-3 polymorphism on specific IgG responses for COVID-19 in older adults seropositive for CMV. METHODS: Blood samples of 25 older adults of both sexes were obtained at three different times: during a micro-outbreak (MO) of SARS-CoV-2 in 2020; eight months after (CURE); and 30 days after the administration of the second dose of ChadOx-1 vaccine (VAC). The specific IgG for both SARS-CoV-2 and CMV antigens, neutralizing antibodies against SARS-CoV-2, and also the polymorphism profile for IFN-lambda-3 (rs12979860 C > T) were assessed. RESULTS: Higher levels of specific IgG for SARS-CoV-2 antigens were found in the MO and VAC than in the CURE time-point. Volunteers with specific neutralizing antibodies against SARS-CoV-2 showed better specific IgG responses for SARS-CoV-2 and lower specific IgG levels for CMV than volunteers without specific neutralizing antibodies. Significant negative correlations between the specific IgG levels for SARS-CoV-2 and CMV were found at the MO time-point, as well as in the group of individuals homozygous for allele 1 (C/C) in the MO time-point and heterozygotes (C/T) in the CURE time-point. CONCLUSION: Our results suggested that both CMV seropositivity and the homozygosis for allele 1 (C/C) in IFN-lambda-3 gene can negatively impact the antibody response to COVID-19 infection and vaccination in older adults.

Adhesion analysis via a tumor vasculature-like microfluidic device identifies CD8+ T cells with enhanced tumor homing to improve cell therapy.

CD8+ T cell recruitment to the tumor microenvironment is critical for the success of adoptive cell therapy (ACT). Unfortunately, only a small fraction of transferred cells home to solid tumors. Adhesive ligand-receptor interactions have been implicated in CD8+ T cell homing; however, there is a lack of understanding of how CD8+ T cells interact with tumor vasculature-expressed adhesive ligands under the influence of hemodynamic flow. Here, the capacity of CD8+ T cells to home to melanomas is modeled ex vivo using an engineered microfluidic device that recapitulates the hemodynamic microenvironment of the tumor vasculature. Adoptively transferred CD8+ T cells with enhanced adhesion in flow in vitro and tumor homing in vivo improve tumor control by ACT in combination with immune checkpoint blockade. These results show that engineered microfluidic devices can model the microenvironment of the tumor vasculature to identify subsets of T cells with enhanced tumor infiltrating capabilities, a key limitation in ACT.

Veno-venous extracorporeal membrane oxygenation in patients with SARS-CoV-2 pneumonia in Brazil: a case series / Oxigenação por membrana extracorpórea venovenosa em pacientes com pneumonia por SARS-CoV-2 no Brasil: série de casos

ABSTRACT Objective: The world has been suffering from the COVID-19 pandemic. Some COVID-19 patients develop severe viral pneumonia, requiring mechanical ventilation and measures to treat refractory hypoxemia, such as a protective ventilation strategy, prone positioning, and the use of veno-venous extracorporeal membrane oxygenation (VV-ECMO). We describe a case series of 30 COVID-19 patients who needed VV-ECMO at the Hospital Alemão Oswaldo Cruz, located in the city of São Paulo, Brazil. Methods: We included all patients who required VV-ECMO due to COVID-19 pneumonia between March of 2020 and June of 2021. Results: Prior to VV-ECMO, patients presented with the following median scores: SOFA score, 11; APPS score, 7; Respiratory ECMO Survival Prediction score, 2; and Murray score, 3.3. The 60-day-in-hospital mortality was 33.3% (n = 10). Conclusions: Although our patients had a highly severe profile, our results were similar to those of other cohort studies in the literature. This demonstrates that VV-ECMO can be a good tool even in a pandemic situation when it is managed in an experienced center.

RESUMO Objetivo: O mundo vem sofrendo com a pandemia de COVID-19. Alguns pacientes com COVID-19 desenvolvem pneumonia viral grave, necessitando ventilação mecânica e medidas para tratar a hipoxemia refratária, como estratégias de ventilação protetora, posição prona e uso de oxigenação por membrana extracorpórea venovenosa (ECMO-VV). Descrevemos uma série de casos de 30 pacientes com COVID-19 que necessitaram de ECMO-VV no Hospital Alemão Oswaldo Cruz, localizado na cidade de São Paulo, Brasil. Métodos: Foram incluídos todos os pacientes que necessitaram de ECMO-VV devido à pneumonia por COVID-19 entre março de 2020 e junho de 2021. Resultados: Antes da ECMO-VV, os pacientes apresentavam as seguintes medianas: escore SOFA de 11; escore APPS de 7; escore Respiratory ECMO Survival Prediction de 2; e escore de Murray de 3,3. A mortalidade hospitalar em 60 dias foi de 33,3% (n = 10). Conclusões: Apesar de nossos pacientes apresentarem um perfil de alta gravidade, nossos resultados foram semelhantes aos de outros estudos de coorte na literatura. Isso demonstra que a ECMO-VV pode ser uma boa ferramenta mesmo em uma situação de pandemia quando administrada em um centro experiente.

Multisystem inflammatory syndrome in children: a case series.

OBJECTIVE: To describe a case series of multisystem inflammatory syndrome in children (MIS-C) in a pediatric tertiary hospital. METHODS: Patients under the age of 18 years who met MIS-C criteria of the Brazilian Ministry of Health (MH) and/or the Royal College of Paediatrics and Child Health (RCPCH) were included. A retrospective analysis was carried out by reviewing medical records and complementary exams. RESULTS: Six pediatric patients with mean age of 126 months were admitted with fever associated with multisystem involvement: all of them had abdominal pain and diarrhea and two underwent appendectomy; 100% had coagulopathy and increased inflammatory markers; 83% had cardiovascular impairment and 60% required vasoactive drugs; 83% had mucocutaneous symptoms and 50% required ventilatory support by invasive mechanical ventilation or non-invasive ventilation. One patient showed coronary artery dilation on echocardiogram. All patients received empiric antibiotic therapies. SARS-CoV-2 IgG testing was positive in five patients. Treatment was performed after excluding infectious causes: five patients (83%) received intravenous immunoglobulin, five patients (83%) pulse methylprednisolone therapy and one (16%) Tocilizumab. One patient died. The average length of stay in Pediatric Intensive Care Unit (PICU) was seven days. CONCLUSIONS: These cases are added to the literature in construction of this emerging condition. Early diagnosis should be considered due to its potential severity.

Turbulence in microfluidics: Cleanroom-free, fast, solventless, and bondless fabrication and application in high throughput liquid-liquid extraction.

This paper addresses an important breakthrough in the deployment of ultra-high adhesion strength microfluidic technologies to provide turbulence at harsh flow rate conditions. This paper is only, to our knowledge, the second reporting on the generation of high flow rate-assisted turbulence in microchannels. This flow solves a crucial bottleneck in microfluidics: the generation of high throughput homogeneous mixings. We focused on the fabrication of bulky polydimethylsiloxane (PDMS) microchips (without any interfaces) rather than the laborious surface modifications that were employed in the first reporting about turbulence-assisted microfluidics. The fabrication is cleanroom-free, simple, low-cost, fast, solventless, and bondless requiring only a laboratory oven. More specifically, our method relies on the shaping of a nylon scaffold, cure of PDMS with embedded nylon, and removal of this scaffold. The scaffold was obtained by manually wrapping nylon threads. The withdrawing out of the scaffold was completed in few seconds using only a plier. Such microchannels endured flow rates of up to 60.0 mL min(-1) with a strikingly low elastic deformation. The importance in producing turbulence into microscale channels was successfully shown in liquid-liquid extractions. The great energy dissipation rate relative to the turbulence created high throughput and efficient extractions in microfluidics for the first time. The residence time was only 0.01 s at 25.0 mL min(-1) (total flow rate of the immiscible phases). In addition, the partition coefficient determined in a single run was similar to that obtained by the conventional batch shake-flask method that was realized in triplicate.

Avaliação microbiológica de queijos coloniais da região de Cascavel - PR

A produção artesanal de queijos é bastante tradicional no Brasil, sendo mais comum no Sul, Sudeste e Centro-Oeste do País, apesar da evolução tecnológica e higienicossanitária na produção de queijos. Além disso, apesar das inúmeras ferramentas elaboradas para garantir a qualidade dos alimentos projeções da ABIQ (Associação Brasileira das Indústrias de Queijo) mostram que apenas 60% da produção nacional de queijos são inspecionadas, sendo os 40% restantes, produzidos e comercializados com qualidade microbiológica e condições higienicossanitárias desconhecidas. Diante dessa situação, o objetivo deste trabalho foi verificar a qualidade microbiológica dos queijos coloniais comercializados na feira livre da cidade de Cascavel,através da observação da incidência Staphylococcus auerus, Salmonella sp. e coliformes 45°C, além de fazer um comparativo com o trabalho"Avaliação Microbiológica de Queijos Coloniais Produzidos no Estado do Paraná" (KOTTWITZ e GUIMARÃES, 2003), para verificar as condições higienicossanitárias e queijos coloniais do interior do estado do Paraná são semelhantes às encontradas em 2001, por Kottwitz e Guimarães.

The artisan cheese production is quite traditional in Brasil, being more common in South, Southeast and Midwest of this country, despite the technological and hygienic production of cheeses. Furthermore, despite numerous tools designed to ensure the quality of food, projections of ABIQ (Brazilian Association of Cheese Industries) show that only 60% of the national production of cheese are inspected, and being the remaining 40% produced and marketed with microbiological quality and unknown higienicossanitárias conditions. Before the situation, the objective of this study was to verify the microbiological quality of the cheeses sold at the free market in Cascavel city, by observation of the incidence of Staphylococcus aureus, Salmonella sp. and coliforms 45ºC, and beyond this, to make a comparison with the work "Evaluation of Microbiological of Colonial Cheese produced in Paraná" (KOTTWITZ and GUIMARÃES, 2003), in order to verify if the hygienic conditions of colonial cheeses in State of Paraná are similar those found in 2001 by Kottwitz and Guimarães.

The effects of cyclosporin A and Heteropterys tomentosa on the rat liver.

Cyclosporin A (CsA) is a widely employed immunosuppressive drug that is associated with several side effects, among then hepatotoxicity. Heteropterys tomentosa is a Brazilian plant efficient in reducing damage caused by CsA on the rat testis and prostate. The aim of this study was to evaluate the effect of CsA and H. tomentosa (administered isolated or simultaneously) on the liver of Wistar rats. The animals were treated daily with water (control), CsA (15 mg/kg/day), H. tomentosa infusion or CsA+H. tomentosa, for 21 or 56 days. The treatments did not alter liver morphology or cause fibrosis. H. tomentosa administered for 21 days increased the number of hepatocyte nuclei and Kupffer cell volumetric proportion. After 56 days of treatment, H. tomentosa administration did not alter the parameters analyzed. Biochemical plasma dosages and liver stereology showed impairment caused by CsA-treatment after 21 days; these results were not observed after 56 days of treatment. The simultaneous treatment with CsA and H. tomentosa for 21 or 56 days did not alleviate nor accentuate CsA hepatic effects. The present study showed that the 21 days treatment with CsA caused more alteration to the liver than the 56 days treatment; this could be related to hepatic recovery after the long term treatment.

Hepatoprotective effect of Arctium lappa root extract on cadmium toxicity in adult Wistar rats.

This study was performed to determine the effects of Arctium lappa (Al) to protect against cadmium damage in the rat liver. Male rats received a single i.p. dose of CdCl2 (1.2 mg/kg body weight (BW)) with or without Al extract administered daily by gavage (300 mg/kg BW) for 7 or 56 days. After 7 days, Al caused plasma transaminase activity to diminish in groups Al (glutamic oxaloacetic transaminase (GOT) and glutamic pyruvic transaminase (GPT)) and CdAl (GPT). After 56 days, GOT and GPT plasma activities were reduced in the Cd group. No alteration in plasma levels of creatinine, total bilirubin, and total protein were observed. GOT liver activity increased in the Cd group. No alteration was observed in superoxide dismutase (SOD), catalase (CAT), glutathione-S-transferase (GST), and malondialdehyde (MDA) dosage. In the Cd group, hepatocyte proportion decreased and sinusoid capillary proportion increased. In the Al and CdAl groups, the nuclear proportion increased and the cytoplasmic proportion decreased. The hepatocyte nucleus density reduced in Cd and increased in the Al group. After 56 days, there was no alteration in the Cd group. In Al and CdAl groups, the nuclear proportion increased without cytoplasmic proportion variation, but the sinusoid capillary proportion was reduced. The hepatocyte nucleus density decreased in the Cd group and increased in the Al and CdAl groups. In conclusion, the liver function indicators showed that A. lappa protected the liver against cadmium toxicity damage.

Naringin inhibits tumor growth and reduces interleukin-6 and tumor necrosis factor α levels in rats with Walker 256 carcinosarcoma.

The flavonoid naringin is a polyphenolic compound that naturally occurs in citrus. Patients with cancer generally present features of malnutrition and cachexia. Levels of the proinflammatory cytokines tumor necrosis factor α (TNF-α) and interleukin-6 (IL-6) are raised in patients with cancer. This study was designed to analyze the in vivo effect of naringin in the therapeutic treatment of rats bearing Walker 256 carcinosarcoma (W256). Rats were treated intraperitoneally with different doses of naringin (10, 25 and 35 mg/kg), for 50 days. At 25 mg/kg, naringin inhibited tumor growth by ~75%. With this treatment, TNF-α and IL-6 levels decreased (p<0.05) in comparison with the control. In addition, two rats presented complete tumor regression. Inhibition of tumor growth, survival increase and the reduction of TNF-α and IL-6 levels in rats bearing W256 treated with naringin strongly suggest that this compound has potential as an anticarcinogenic drug.

Inhibition of tumor growth by quercetin with increase of survival and prevention of cachexia in Walker 256 tumor-bearing rats.

Quercetin, a flavonoid abundantly present in fruit, vegetables, wine and tea, has revealed several properties such as antioxidant, antiproliferative and anticancer. Cachexia is a poorly understood syndrome present in already compromised cancer patients, decreasing the quality of life and increasing mortality. Many studies have been performed in an attempt to discover an effective treatment for cachexia, but none of the tested therapies has fulfilled expectations. The objective of the present study was to analyze the effect of quercetin in the therapeutic treatment of cachexia and reversion of tumor growth in rats bearing Walker 256 carcinosarcoma (W256). Rats bearing W256 were treated daily with I.P. quercetin injections, at different doses (10, 15, 25 and 35 mg/kg). The results show that 10 mg/kg quercetin inhibited tumor growth by about 50% (ED(50)) when compared with controls (CTR). Moreover, two animals of this group presented complete tumor regression. Matrix metalloproteinase-2 (MMP-2) activity and vascular endothelial growth factor (VEGF) expression decreased in rats bearing W256 treated with 10 mg/kg quercetin when compared with CTR. Thus, the inhibition of tumor growth, survival increase, decrease of MMP-2 and VEGF levels and reduction of cachexia in animals treated with quercetin strongly support the anticancer function of this flavonoid.

Assessment of deep venous thrombosis by grayscale median analysis of ultrasound images.

AIM: This study aimed to determine whether grayscale median (GSM) ultrasound (US) analysis could differentiate acute from subacute femoropopliteal deep venous thrombosis (DVT) classified based on (a) the time of symptoms onset and (b) sonographers' evaluation of US images. METHODS: Patients who had US because of suspected DVT were included. Patients with history of previous DVT, with normal US results or US with findings of isolated calf thrombosis were excluded. A total of 128 venous segments of 63 patients were analyzed. Thrombosed veins and adjacent arteries images were recorded. Thrombi GSM were calculated. Grayscale median was compared with (a) the time of symptoms onset and (b) US image thrombi classification as acute or subacute done by 2 sonographers blinded to patient information. Statistical analysis included GSM Pearson intraobserver variability and t test comparisons of GSM means for (a) short versus long onset periods and (b) acute versus subacute US class, χ comparison between onset history and US class, creation of GSM accuracy, predictive values, and sensitivity curves for acute versus subacute DVT. RESULTS: Intraobserver variability correlation varied between 0.92 and 0.95. Thrombus mean GSM was 23 ± 12 versus 31 ± 16 if symptoms onset was shorter or longer than 2 weeks (P = 0.004). Mean GSM was 20 ± 11 versus 35 ± 14 for acute versus subacute thrombi (P < 0.001). Acute thrombi correlated with onset shorter than 2 weeks (P < 0.001). Grayscale median less than 13, equal to 22, or 33 or less had 85% positive predictive value (17/20), 77% accuracy (98/128), and 90% sensitivity (53/59) to differentiate acute from subacute thrombi. CONCLUSIONS: Grayscale median of US images documenting femoropopliteal DVT correlated with (a) the time between symptoms onset and US examination and (b) the sonographers' interpretation of acute versus subacute thrombosis.