Development of a new assay for quantification of parasite load of intracellular Leishmania sp. in macrophages using flow cytometry.

Finding novel methodologies that enhance the precision, agility, and standardization of drug discovery is crucial for studying leishmaniasis. The slide count is the technique most used to assess the leishmanicidal effect of a given drug in vitro. Despite being consolidated in the scientific environment, it presents several difficulties in its execution, assessment, and results. In addition to being laborious, this technique takes time, both for the preparation of the material for analysis and for the counting itself. Our research group suggests a fresh approach to address this requirement, which involves utilizing nuclear labeling with propidium iodide and flow cytometry to determine the quantity of Leishmania sp. parasites present in macrophages in vitro. Our results show that the fluorescence of infected samples increases as the infection rate increases. Using Pearson's Correlation analysis, it was possible to establish a correlation coefficient (Pearson r = 0.9473) that was strongly positive, linear, and directly proportional to the fluorescence and infection rate variables. Thus, it is possible to infer a mathematical equation through linear regression to estimate the number of parasites in each sample using the Relative Fluorescence Units (RFU) values. This new methodology opens space for the possibility of using this methodological resource in the in vitro quantification of Leishmania in macrophages.

Neglect scoring modifications in the National Institutes of Health Stroke Scale improve right hemisphere stroke lesion volume prediction.

BACKGROUND: The National Institutes of Health Stroke Scale (NIHSS) does not equitably assess stroke severity in the two cerebral hemispheres. By attributing a maximum of two points for neglect and seven for language, it undervalues right hemisphere deficits. We aimed to investigate if NIHSS equally predicts right hemisphere lesion volumes in patients with and without neglect, and if a modification of the neglect scoring rules could increase its predictive capacity. METHODS: We analyzed a prospective cohort of acute right middle cerebral artery ischemic stroke patients. First, we calculated the correlation between NIHSS scores and lesion volume and analyzed the partial correlation of neglect. Then, we applied different modifications in the neglect scoring rules and investigated how they interfered with lesion volume predictive capacity. RESULTS: A total of 162 ischemic stroke patients were included, 108 with neglect and 54 without. The correlation between lesion volume and NIHSS was lower in patients with neglect (r = 0.540 vs. r = 0.219, p = 0.004) and neglect was a statistically significant covariate in the partial correlation analysis between NIHSS and lesion volume (p = 0.017). With the neglect score tripled and with the duplication or triplication of all neglect modalities, the correlation was significantly higher than with the standard NIHSS (p = 0.043, p = 0.005, p = 0.001, respectively). With these modifications, neglect was no longer a significant covariable in the partial correlation between lesion volume and NIHSS. CONCLUSION: A modification of NIHSS neglect scoring might improve the scale's capacity to predict lesion volume.

Biochemical characterization of an esterase from Thermobifida fusca YX with acetyl xylan esterase activity.

BACKGROUND: Esterases (EC 3.1.1.X) are enzymes that catalyze the hydrolysis ester bonds. These enzymes have large potential for diverse applications in fine industries, particularly in pharmaceuticals, cosmetics, and bioethanol production. METHODS AND RESULTS: In this study, a gene encoding an esterase from Thermobifida fusca YX (TfEst) was successfully cloned, and its product was overexpressed in Escherichia coli and purified using affinity chromatography. The TfEst kinetic assay revealed catalytic efficiencies of 0.58 s-1 mM-1, 1.09 s-1 mM-1, and 0.062 s-1 mM-1 against p-Nitrophenyl acetate, p-Nitrophenyl butyrate, and 1-naphthyl acetate substrates, respectively. Furthermore, TfEst also exhibited activity in a pH range from 6.0 to 10.0, with maximum activity at pH 8.0. The enzyme demonstrated a half-life of 20 min at 70 °C. Notably, TfEst displayed acetyl xylan esterase activity as evidenced by the acetylated xylan assay. The structural prediction of TfEst using AlphaFold indicated that has an α/ß-hydrolase fold, which is consistent with other esterases. CONCLUSIONS: The enzyme stability over a broad pH range and its activity at elevated temperatures make it an appealing candidate for industrial processes. Overall, TfEst emerges as a promising enzymatic tool with significant implications for the advancement of biotechnology and biofuels industries.

Preventing occludin tight-junction disruption via inhibition of microRNA-193b-5p attenuates viral load and influenza-induced lung injury.

Virus-induced lung injury is associated with loss of pulmonary epithelial-endothelial tight junction integrity. While the alveolar-capillary membrane may be an indirect target of injury, viruses may interact directly and/or indirectly with miRs to augment their replication potential and evade the host antiviral defense system. Here, we expose how the influenza virus (H1N1) capitalizes on host-derived interferon-induced, microRNA (miR)-193b-5p to target occludin and compromise antiviral defenses. Lung biopsies from patients infected with H1N1 revealed increased miR-193b-5p levels, marked reduction in occludin protein, and disruption of the alveolar-capillary barrier. In C57BL/6 mice, the expression of miR-193b-5p increased, and occludin decreased, 5-6 days post-infection with influenza (PR8). Inhibition of miR-193b-5p in primary human bronchial, pulmonary microvascular, and nasal epithelial cells enhanced antiviral responses. miR-193b-deficient mice were resistant to PR8. Knockdown of occludin, both in vitro and in vivo, and overexpression of miR-193b-5p reconstituted susceptibility to viral infection. miR-193b-5p inhibitor mitigated loss of occludin, improved viral clearance, reduced lung edema, and augmented survival in infected mice. Our results elucidate how the innate immune system may be exploited by the influenza virus and how strategies that prevent loss of occludin and preserve tight junction function may limit susceptibility to virus-induced lung injury.

Erosive Tooth Wear, Wine Intake, and Genetic Variation in COMT and MMP2.

INTRODUCTION: Erosive tooth wear (ETW) is a multifactorial condition of increasing prevalence in the younger population. This study aimed to explore the association between different ETW phenotypes with MMP2 and COMT single-nucleotide variants, and selected environmental factors. METHODS: Saliva samples, erosive wear and dental caries experience data, and dietary/behavioral information from 16-18-year-old patients (n= 747) were used. Genotypes were obtained and phenotypes were further analyzed considering diet and behavioral data, using logistic regression as implemented in PLINK, with an alpha of 0.05. RESULTS: When comparing individuals' ETW-free with those with mild ETW, an association was found with COMT rs6269 (p = 0.02). The comparison between ETW-free individuals with individuals with severe ETW also showed an association with COMT rs6269 under the recessive model (p = 0.03). Logistic regression showed that in the presence of less common alleles of MMP2 rs9923304 and COMT rs6269, ETW were more likely to occur when individuals drank wine. The GG genotype of COMT rs6269 was associated with the presence of lower (p = 0.02) and higher (p = 0.02) caries experience when individuals with ETW only in enamel were compared with individuals with ETW involving dentin. CONCLUSION: The results support a role of genes in ETW, with wine consumption being identified as a significant modulator, suggesting that gene-environment interactions may contribute to the development of erosive tooth wear.

Cannabidiol-loaded microparticles embedded in a porous hydrogel matrix for biomedical applications.

In this study, poly (lactic-co-glycolic acid) (PLGA) microparticles loaded with cannabidiol (CBD) were synthesized (PLGA@CBD microparticles) and embedded up to 10 wt% in a chondroitin sulfate/polyvinyl alcohol hydrogel matrix. In vitro chemical, physical, and biological assays were carried out to validate the potential use of the modified hydrogels as biomaterials. The microparticles had spherical morphology and a narrow range of size distribution. CBD encapsulation efficiency was around 52%, loading was approximately 50%. Microparticle addition to the hydrogels caused minor changes in their morphology, FTIR and thermal analyses confirmed these changes. Swelling degree and total porosity were reduced in the presence of microparticles, but similar hydrophilic and degradation in phosphate buffer solution behaviors were observed by all hydrogels. Rupture force and maximum strain at rupture were higher in the modified hydrogels, whereas modulus of elasticity was similar across all materials. Viability of primary human dental pulp cells up to 21 days was generally not influenced by the addition of PLGA@CBD microparticles. The control hydrogel showed no antimicrobial activity against Staphylococcus aureus, whereas hydrogels with 5% and 10% PLGA@CBD microparticles showed inhibition zones. In conclusion, the PLGA@CBD microparticles were fabricated and successfully embedded in a hydrogel matrix. Despite the hydrophobic nature of CBD, the physicochemical and morphological properties were generally similar for the hydrogels with and without the CBD-loaded microparticles. The data reported in this study suggested that this original biomaterial loaded with CBD oil has characteristics that could enable it to be used as a scaffold for tissue/cellular regeneration.

Sanguinarine: an alkaloid with promising in vitro and in vivo antiparasitic activity against different developmental stages of Schistosoma mansoni and in silico pharmacokinetic properties (ADMET).

The objective of the study was to evaluate the in vitro and in vivo schistosomicidal activity of sanguinarine (SA) on Schistosoma mansoni and its in silico pharmacokinetic parameters. ADMET parameters and oral bioavailability were evaluated using the PkCSM and SwissADME platforms, respectively. The activity of SA in vitro, at the concentrations of 1.0-25 µM, was analyzed through the parameters of motility, mortality, and cell viability of the worms at intervals of 3-24 h. Mice were infected with cercariae and treated by gavage with SA (5 mg/kg/day, in a single dose or two doses of 2.5 mg/kg every 12 h for 5 consecutive days) on the 1st (skin schistosomula), 14th (pulmonary schistosomula), 28th (young worms), and 45th (adult worms) days after infection. In vitro and in vivo praziquantel was the control. In vitro, SA showed schistosomicidal activity against schistosomula, young worms, and couples; with total mortality and reduced cell viability at low concentrations and incubation time. In a single dose of 5 mg/kg/day, SA reduces the total worm load by 47.6%, 54%, 55.2%, and 27.1%, and female worms at 52.0%, 39.1%, 52.7%, and 20.2%, respectively, results which are similar to the 2.5 mg/kg/day dose. SA reduced the load of eggs in the liver, and in histopathological and histomorphometric analyses, there was a reduction in the number and volume of hepatic granulomas, which exhibited less inflammatory infiltrate. SA has promising in vitro and in vivo schistosomicidal activity against different developmental stages of S. mansoni, in addition to reducing granulomatous liver lesions. Furthermore, in silico, SA showed good predictive pharmacokinetic ADMET profiles.

Kisspeptin-10 Improves Testicular Redox Status but Does Not Alter the Unfolded Protein Response (UPR) That Is Downregulated by Hypothyroidism in a Rat Model.

Hypothyroidism compromises the testicular redox status and is associated with reduced sperm quality and infertility in men. In this regard, studies have demonstrated the antioxidant potential of kisspeptin in reproductive and metabolic diseases. In this study, we evaluate the effects of kisspeptin-10 (Kp10) on the testicular redox, as well as mediators of the unfolded protein response (UPR) in adult rats with hypothyroidism. Adult male Wistar rats were randomly separated into the Control (n = 15), Hypo (n = 13) and Hypo + Kp10 (n = 14) groups, and hypothyroidism was induced with 6-propyl-2-thiouracil (PTU) for three months. In the last month, half of the hypothyroid animals received Kp10. Testis samples were collected for enzymatic, immunohistochemical and/or gene evaluation of mediators of oxidative stress (TBARs, lipid hydroperoxides (LOOH), ROS, peroxynitrite, SOD, CAT and GPX), endoplasmic reticulum stress (GRP78, ATF6, PERK, CHOP, HO-1 and sXBP1) and antiapoptocytes (BCL-2). Hypothyroidism increased apoptosis index, TBARS and LOOH concentrations, and reduced testicular gene expression of Sod1, Sod2 and Gpx1, as well as the expression of Grp78, Atf6, Ho1 and Chop. Treatment with Kp10, in turn, reduced testicular apoptosis and the production of peroxynitrite, while increased SOD1 and GPX ½ expression, and enzymatic activity of CAT, but did not affect the lower expression of UPR mediators caused by hypothyroidism. This study demonstrated that hypothyroidism causes oxidative stress and dysregulated the UPR pathway in rat testes and that, although Kp10 does not influence the low expression of UPR mediators, it improves the testicular redox status, configuring it as an important antioxidant factor in situations of thyroid dysfunction.

Tyrosine-Mutant AAV8 Vector Mediated Efficient and Safe Gene Transfer of Pigment Epithelium-Derived Factor to Mouse Lungs.

BACKGROUND/AIMS: Recombinant adeno-associated viruses (rAAV) are an important tool for lung targeted gene therapy. Substitution of tyrosine with phenylalanine residues (Y-F) in the capsid have been shown to protect the AAV vector from ubiquitin/proteasome degradation, increasing transduction efficiency. We tested the mutant Y733F-AAV8 vector for mucus diffusion, as well as the safety and efficacy of pigment epithelium-derived factor (PEDF) gene transfer to the lung. METHODS: For this purpose, Y733F-AAV8-PEDF (1010 viral genome) was administered intratracheally to C57BL/6 mice. Lung mechanics, morphometry, and inflammation were evaluated 7, 14, 21, and 28 days after injection. RESULTS: The tyrosine-mutant AAV8 vector was efficient at penetrating mucus in ex vivo assays and at transferring the gene to lung cells after in vivo instillation. Increased levels of transgene mRNA were observed 28 days after vector administration. Overexpression of PEDF did not affect in vivo lung parameters. CONCLUSION: These findings provide a basis for further development of Y733F-AAV8-based gene therapies for safe and effective delivery of PEDF, which has anti-angiogenic, anti-inflammatory and anti-fibrotic activities and might be a promising therapy for lung inflammatory disorders.

Technological aspects and health effects of hydrolyzed collagen and application in dairy products.

With the rise of a consumer market increasingly concerned with food and healthy lifestyle habits, the search for functional products has increased in the last years. In this context, dairy products are relevant since they are already included in the consumer's diet. Furthermore, hydrolyzed collagen stands out among products with bioactive action, as it promotes the reduction of the incidence of arthritis, osteoporosis, hypertension, obesity, and premature aging and contains healing, antioxidant and antimicrobial properties. In addition to health benefits, the addition of these ingredients to dairy products can influence physical, chemical, rheological, microbiological, and sensory characteristics, such as: decreased syneresis and improved texture of fermented milks; viscosity increase in dairy beverage; increased proteolytic activity in cheeses; and increasing the viability of probiotics, without significantly altering the quality standards of the legislation. Despite the benefits described, more studies are needed to evaluate these effects in different dairy products.

Significant increased bacterial contamination with endoscope overnight and weekend storage times.

BACKGROUND AND AIM: Forced-air drying (FAD) cabinets are recommended for storage of reprocessed endoscopes, but financial constraints prevent their universal application. The study aimed to determine bacterial contamination in flexible gastroscopes (FG) channels after storage, in a cabinet with filtered air and UV lights, but without FAD. METHODS: Eight FG in clinical use in an endoscopy service of a large Brazilian hospital were sampled: immediately "Time zero" (N = 50), 12 h "Time 1" (N = 25), and 60 h "Time 2" (N = 25) after reprocessing. Following a flush-brush-flush of channels, 40-mL sterile water and 3 cm of the brush were collected. Each sample was divided, filtered onto two 0.22-µm membranes, and incubated in media without or with disinfectant neutralizer. Automated method was used for identification and antibiotic resistance test of isolated bacteria. RESULTS: Bacterial contamination in times "1" and "2" was 5.9 and 16.1 times greater than that of "Time zero," respectively. Number of positive cultures in media with and without neutralizer was similar at times "1" and "2," while media with neutralizer produced more positive cultures at "Time zero." Most bacteria isolated at "Time 2" were Gram-negative rods (52.3%) and showed resistance to one or more antibiotics (65%). CONCLUSION: Bacterial contamination was detected on reprocessed FG stored in non-FAD cabinets overnight (12 h) and increased with longer storage time (60 h). The contamination source is likely to be bacteria in biofilm which multiply in the absence of FAD. Evidence-based criteria should be available for storage time according to the cabinet available.

Polymorphisms associated with oral clefts as potential markers for oral pre and malignant disorders.

OBJECTIVE: To investigate whether genes in the Wnt pathway, which have been previously associated with both oral clefts and oral squamous cell carcinoma, are also associated with oral potentially malignant disorders (leukoplakia, erythroplakia and lichen planus). MATERIALS AND METHODS: Case-control study: Dataset consisted of clinical information linked to DNA samples from affected subjects diagnosed with oral potential malignant disorders and oral cancer and their matched controls. Individual samples, clinical history, and potential risk factors were obtained through the Dental Registry and DNA Repository project of the School of Dental Medicine, University of Pittsburgh. The rs1533767 (WNT11), rs9879992 (GSK3B), and rs3923087 (AXIN2) were tested. After genomic DNA had been extracted, genotyping was performed blindly to clinical diagnosis status. Representation of genotypes and alleles in affected subjects in comparison to the unaffected individuals was determined using PLINK. Additional analysis was performed to investigate associations between environmental (socioeconomic/lifestyle) risk factors and the oral pathologies studied using STATA. RESULTS: Two of the SNPs tested (rs9879992 in GSK3B and rs3923087 in AXIN2) were statistically, significantly associated with the pathologies studied (p = 0.039 and 0.038, respectively). CONCLUSION: Single-nucleotide polymorphisms in genes in the Wnt pathway were associated with oral potentially malignant disorders.

Salivary biomarkers to evaluate psychological disorders in oral lichen planus: A systematic review with meta-analysis.

OBJECTIVE: To analyze the use of salivary biomarkers to identify psychological disorders (anxiety, depression, and stress) in individuals with oral lichen planus (OLP). MATERIALS AND METHODS: The search was made in Medline/Pubmed, Scopus, Web of Science, Virtual Health Library, ProQuest, and by cross-reference. Original case-control, cross-sectional, and cohort studies, in human subjects (age ≥18 years) with clinical and histopathological diagnosis, with or without the use of psychometric scales, published until June 2021, were included. Random-effects meta-analyses, subgroup analyses, and sensitivity were performed. Publication bias was measured using the Egger test and visual inspection of the funnel plot. RESULTS: A total of 67 articles were found, of which 12 were included in the meta-analysis. Among the different biomarkers identified, it was only possible to perform a meta-analysis for cortisol. The mean difference between salivary cortisol levels in patients with OLP compared to controls was 3.43 ng/ml (95% CI: 1.20-5.65) with I2  = 98.9%. Studies considered outliers were removed and based on sensitivity analyses, results did not change, therefore, being stable. CONCLUSION: The studies analyzed demonstrated that cortisol was the most quantified biomarker to identify psychological disorders; however, DHEA, IgA, adiponectin, and α-amylase were also evaluated.

[Addressing HIV/AIDS and syphilis in Venezuelan migrant women from the perspective of health managers in the North of BrazilManejo de la infección por el VIH/sida y la sífilis en mujeres migrantes venezolanas desde la perspectiva de los gestores de salud en el norte de Brasil]. / Enfrentamento de HIV/aids e sífilis em mulheres venezuelanas migrantes na perspectiva de gestores de saúde no Norte do Brasil.

Objective: To identify the perception of health managers regarding the actions taken and the challenges encountered in addressing HIV and syphilis in Venezuelan migrant women in Brazil. Method: This descriptive-exploratory study using a qualitative approach was conducted from January to March 2021 in the municipalities of Boa Vista (state of Roraima) and Manaus (state of Amazonas). Audio interviews with the participants were transcribed in full and examined using thematic content analysis. Results: Ten managers were interviewed (five in Boa Vista and five in Manaus). Content analysis identified the following domains and themes: available infrastructure for diagnosis and treatment of AIDS and syphilis - access, availability of medical appointments /waiting list, training of health teams, and psychosocial support; challenges faced by Venezuelan women - language, documentation issues, and frequent address changes; and strategies and actions adopted and expectations for addressing HIV/AIDS and syphilis in the context of migration. Conclusions: Despite the care provided to Venezuelan women in Brazil - guaranteed by the universal nature of the Brazilian health system - language and lack of documentation remain as barriers. Given the absence of action plans and future planning for the care of migrant women with HIV or syphilis in municipalities, it is important to develop public policies aimed at minimizing the difficulties faced by this population.

Objetivo: Determinar la percepción de los gestores de salud sobre las medidas adoptadas y los desafíos encontrados para manejar la infección por el VIH y la sífilis en mujeres migrantes venezolanas en Brasil. Métodos: Este estudio descriptivo y exploratorio, con enfoque cualitativo, se realizó de enero a marzo del 2021 en los municipios de Boa Vista (estado de Roraima) y Manaos (estado de Amazonas). La grabación de las entrevistas de los participantes se transcribió en su totalidad. El análisis se basó en la técnica de evaluación temática del contenido. Resultados: Se hicieron entrevistas a diez gestores (cinco en Boa Vista y cinco en Manaos). En el análisis del contenido se observaron los siguientes ejes y temas: la estructura disponible para el diagnóstico y tratamiento del sida y de la sífilis, con inclusión de acceso, cupos de atención, filas de espera y formación de equipos de salud y apoyo psicosocial; los desafíos enfrentados por las mujeres venezolanas, como idioma, problemas de documentación y frecuencia de cambio de domicilio; las estrategias y medidas adoptadas y las expectativas para enfrentar la infección por el VIH/sida y la sífilis en el marco de la migración. Conclusiones: A pesar de las medidas de acogida a las mujeres migrantes venezolanas, garantizadas por la universalidad del sistema de salud brasileño, todavía existen algunas barreras como el idioma y la falta de documentación. Ante la falta de planes de acción y planificación futura de la atención a las mujeres migrantes portadoras del VIH o con sífilis en los municipios, es importante formular políticas públicas con el fin de reducir las dificultades enfrentadas por este grupo de la población.

[Barriers and facilitators to confronting HIV/aids and syphilis experienced by Venezuelan women living in BrazilFactores facilitadores y barreras que tienen las mujeres venezolanas residentes en Brasil respecto de la infección por el VIH/sida y la sífilis]. / Barreiras e facilitadores do enfrentamento de HIV/aids e sífilis por venezuelanas residentes no Brasil.

Objective: To describe the perception of Venezuelan women regarding access to health care, diagnosis, and treatment of HIV/aids and syphilis in Brazil. Method: This is a descriptive, exploratory study employing a qualitative approach, performed from February to May 2021 in the municipalities of Manaus, state of Amazonas, and Boa Vista, state of Roraima. The interviews with participants were fully transcribed, with identification of themes based on content analysis. Results: Forty women were interviewed (20 in Manaus and 20 in Boa Vista). Following transcription and translation of the accounts, two analytical categories were identified: barriers to healthcare access, with four subcategories - language, cost, adverse drug reactions, and COVID-19 pandemic; and facilitators of healthcare access, again with four subcategories - Unified Health System (SUS), National Policy of Comprehensive Women's Health, National Social Assistance Policy, and relationship between healthcare professionals and SUS users. Conclusion: The results showed the need to design strategies to mitigate the difficulties faced by migrant women from Venezuela living in Brazil regarding the diagnosis and treatment of HIV/aids and syphilis, going beyond the healthcare support guaranteed by law.

Objetivo: Describir la percepción de las mujeres venezolanas sobre el acceso a los servicios de salud, al diagnóstico y al tratamiento de la infección por el VIH/sida y la sífilis en Brasil. Métodos: Se trata de un estudio descriptivo y exploratorio, con enfoque cualitativo, realizado entre febrero y mayo del 2021 en los municipios de Manaos, estado de Amazonas, y Boa Vista, estado de Roraima. Las entrevistas con las participantes se transcribieron en su totalidad, y se exploraron los puntos de interés según el análisis del contenido. Resultados: Se entrevistaron 40 mujeres (20 en Manaos y 20 en Boa Vista). A partir de la transcripción y la traducción de las conversaciones, se establecieron dos categorías para el análisis del contenido: las barreras de acceso a los servicios de salud (subcategorías: idioma, costos relacionados con la salud, reacciones adversas a los medicamentos y pandemia de COVID-19); y los factores facilitadores del acceso (cuatro subcategorías: Sistema Único de Salud, Política nacional de Atención Integral a la Salud de la Mujer, Política Nacional de Asistencia Social y relación entre los profesionales de salud y las usuarias del Sistema Único de Salud). Conclusión: Los resultados mostraron la necesidad de formular estrategias para mitigar las dificultades que enfrentan las mujeres migrantes de Venezuela residentes en Brasil en relación con el diagnóstico y el tratamiento de la infección por el VIH/sida y la sífilis, más allá de la protección de la salud garantizada por la ley.

Coast-or Inland Residence and Differences in the Occurrence of Cleft Lip and Cleft Palate.

OBJETIVE: To analyze if differences in lifestyle and environment between coastal and inland areas are associated with differences in frequency of orofacial cleft types. DESIGN: Populational cross-sectional study. SETTING: All live borns with orofacial cleft registered at Brazilian Live Birth Information System between 1999 and 2020. PARTICIPANTS: 33,699 live borns with orofacial cleft. INTERVENTION: Data from borns with orofacial cleft were collected at Brazilian Live Birth Information System. MAIN OUTCOME MEASURE: Differences in frequencies between the cleft types and covariates were determined using chi-square. Bivariate analysis was done to obtain the prevalence ratio of types of clefts by geographic origin. Multiple logistic regression analysis was used to determine adjusted odds ratios, controlling for covariates, establishing a significance level of p value <0.05. RESULTS: The frequency of cleft types was statistically significant different according to geographic origin (inland x coast). For syndromic clefts, the prevalence ratio for cleft lip with/without palate was 3.6 times higher inland (p value = 0.000). Regarding non-syndromics, the prevalence ratio for cleft lip with/without palate was two times higher inland (p value = 0.000). Logistic regression suggested cleft lip with/without palate was 6.33 more likely to occur in inland regions (p value = 0.000). CONCLUSION: Geographic origin was associated with the type of cleft in Brazil, with a higher prevalence of cleft lip with/without palate in inland areas, compared to cleft palate, which was higher in the coast.

LioNeo project: a randomised double-blind clinical trial for nutrition of very-low-birth-weight infants.

We assessed the effectiveness of lyophilised banked human milk (HM) as a fortifier to feed very-low-birth-weight infants (VLBWI). This study aimed to evaluate the safety and tolerability of HM with HM lyophilisate as an additive compared with the standard additive (cows' milk protein). In this phase I double-blind randomised controlled clinical trial, set in the intensive and intermediate care units of a tertiary hospital, forty VLBWI were enrolled and allocated into two groups: HM plus HM lyophilisate (LioNeo) or HM plus commercial additive (HMCA). The inclusion criteria were preterm infants, birth weight 750-1500 g, small or adequate for gestational age, exclusively receiving donor HM, volume ≥ 100 ml/kg per d and haemodynamically stable. Participants were followed up for 21 consecutive days. The primary outcome measures were necrotising enterocolitis (NEC), late-onset sepsis (LOS), death, gastrointestinal (GI) bleeding or perforation, diarrhoea, regurgitation, vomiting and abdominal distension. The LioNeo and HMCA groups had similar weights at baseline. The regression models showed no differences between the groups in terms of the primary outcomes. Diarrhoea, GI perforation, NEC and LOS were absent in the LioNeo group (one LOS and one NEC in the HMCA group). Multiple regression analysis with the total volume of milk as a covariate did not show significant differences. The lyophilisation of donor HM was considered safe and tolerable for use in stable haemodynamically VLBWI.

Could combination chemotherapy be more effective than monotherapy in the treatment of visceral leishmaniasis? A systematic review of preclinical evidence.

From a systematic review framework, we assessed the preclinical evidence on the effectiveness of drug combinations for visceral leishmaniasis (VL) treatment. Research protocol was based on the PRISMA guideline. Research records were identified from Medline, Scopus and Web of Science. Animal models, infection and treatment protocols, parasitological and immunological outcomes were analysed. The SYRCLE's (SYstematic Review Center for Laboratory Animal Experimentation) toll was used to evaluate the risk of bias in all studies reviewed. Fourteen papers using mice, hamster and dogs were identified. Leishmania donovani was frequently used to induce VL, which was treated with 23 drugs in 40 different combinations. Most combinations allowed to reduce the effective dose, cost and time of treatment, in addition to improving the parasitological control of Leishmania spp. The benefits achieved from drug combinations were associated with an increased drug's half-life, direct parasitic toxicity and improved immune defences in infected hosts. Selection, performance and detection bias were the main limitations identified. Current evidence indicates that combination chemotherapy, especially those based on classical drugs (miltefosine, amphotericin B antimony-based compounds) and new drugs (CAL-101, PAM3Cys, tufisin and DB766), develops additive or synergistic interactions, which trigger trypanocidal and immunomodulatory effects associated with reduced parasite load, organ damage and better cure rates in VL.

Religious/spiritual coping, symptoms of depression, stress, and anxiety in caregivers of children and adolescents with type 1 diabetes.

BACKGROUND: It was verified the level of religiosity and spirituality, and symptoms of depression, stress, and anxiety of caregivers of children and adolescents with type 1 diabetes, and its interference in glycaemic control. METHODS: Socio-economic and demographic data were collected from caregivers of 59 children and adolescents with type 1 diabetes and obtained dosages of fasting glucose (FG); postprandial glycaemia (PPG); fructosamine (FRUTO); and HbA1c, as well as the glycaemic variability-∆HbA1c. Levels of religiosity were obtained by the DUREL scale; the use of religious/spiritual coping was verified by the SRCOPE-Brief scale. Symptoms of depression, anxiety, and stress were analysed by the DASS-21 scale. The correlations between the variables were analysed by the Pearson coefficient, with significance at 5% level. RESULTS: Inverse correlations were observed between caregivers' schooling with PPG (r = -0.30; P = 0.002) and FRUTO (r = -0.34; P = 0.008) and between family income and FRUTO (r = -0.37; P = 0.004). Direct, moderate correlations were observed between negative religious/spiritual coping (NSRCOPE) with symptoms of depression (r = 0.588; P < 0.0001), stress (r = 0.500; P < 0.0001), and anxiety (r = 0.551; P < 0.0001). CONCLUSION: The direct association between NSRCOPE with symptoms of depression, stress, and anxiety was the highlighted item in the present study emphasizing the need for greater attention to the emotional health of informal caregivers of children and adolescents with DM1. It is necessary to reflect on the religious/spiritual support especially for the main caregiver of children and adolescents with DM1.

Polarization of Microglia and Its Therapeutic Potential in Sepsis.

Sepsis is a life-threatening organ dysfunction caused by a dysregulated host response to infection, leaving the inflammation process without a proper resolution, leading to tissue damage and possibly sequelae. The central nervous system (CNS) is one of the first regions affected by the peripheral inflammation caused by sepsis, exposing the neurons to an environment of oxidative stress, triggering neuronal dysfunction and apoptosis. Sepsis-associated encephalopathy (SAE) is the most frequent sepsis-associated organ dysfunction, with symptoms such as deliriums, seizures, and coma, linked to increased mortality, morbidity, and cognitive disability. However, the current therapy does not avoid those patients' symptoms, evidencing the search for a more optimal approach. Herein we focus on microglia as a prominent therapeutic target due to its multiple functions maintaining CNS homeostasis and its polarizing capabilities, stimulating and resolving neuroinflammation depending on the stimuli. Microglia polarization is a target of multiple studies involving nerve cell preservation in diseases caused or aggravated by neuroinflammation, but in sepsis, its therapeutic potential is overlooked. We highlight the peroxisome proliferator-activated receptor gamma (PPARγ) neuroprotective properties, its role in microglia polarization and inflammation resolution, and the interaction with nuclear factor-κB (NF-κB) and mitogen-activated kinases (MAPK), making PPARγ a molecular target for sepsis-related studies to come.