The human volatilome meets cancer diagnostics: past, present, and future of noninvasive applications.

BACKGROUND: Cancer is a significant public health problem, causing dozens of millions of deaths annually. New cancer screening programs are urgently needed for early cancer detection, as this approach can improve treatment outcomes and increase patient survival. The search for affordable, noninvasive, and highly accurate cancer detection methods revealed a valuable source of tumor-derived metabolites in the human metabolome through the exploration of volatile organic compounds (VOCs) in noninvasive biofluids. AIM OF REVIEW: This review discusses volatilomics-based approaches for cancer detection using noninvasive biomatrices (breath, saliva, skin secretions, urine, feces, and earwax). We presented the historical background, the latest approaches, and the required stages for clinical validation of volatilomics-based methods, which are still lacking in terms of making noninvasive methods available and widespread to the population. Furthermore, insights into the usefulness and challenges of volatilomics in clinical implementation steps for each biofluid are highlighted. KEY SCIENTIFIC CONCEPTS OF REVIEW: We outline the methodologies for using noninvasive biomatrices with up-and-coming clinical applications in cancer diagnostics. Several challenges and advantages associated with the use of each biomatrix are discussed, aiming at encouraging the scientific community to strengthen efforts toward the necessary steps to speed up the clinical translation of volatile-based cancer detection methods, as well as discussing in favor of (i) hybrid applications (i.e., using more than one biomatrix) to describe metabolite modulations that can be "cancer volatile fingerprints" and (ii) in multi-omics approaches integrating genomics, transcriptomics, and proteomics into the volatilomic data, which might be a breakthrough for diagnostic purposes, onco-pathway assessment, and biomarker validations.

[Obesity sociodemographic and behavioral factors: a longitudinal study]. / Fatores sociodemográficos e comportamentais da obesidade: um estudo longitudinal.

This study aimed to investigate the association between sociodemographic and behavioral factors and obesity from 2014 to 2021 in adults in São Paulo city, Brazil. A prospective study was carried out with 1,241 adults aged 18 years or above who lived in São Paulo and participated in the São Paulo Health Survey cohort (ISA) - Physical Activity and Environment. The outcome was obesity (yes/no), classified by body mass index and specific cut-off points for each age group. The exposure variables included sex, age, education, skin color, marital status, regional health coordination, physical activity in the four domains, and sedentary behaviors. Multilevel logistic regression models were used for longitudinal analysis. This study found a significant increase (27.7%) in the prevalence of obesity (from 22.6% to 28.9%). People who practiced at least 150 minutes of leisure-time physical activity per week (OR = 0.44; 95%CI: 0.26; 0.76), between 10 and 150 minutes per week of commuting physical activity (OR = 0.49; 95%CI: 0.30; 0.80), and had no partner (OR = 0.47; 95%CI: 0.28; 0.78) were less likely to be obese. People aged from 40 to 59 years (OR = 5.00; 95%CI: 2.02; 12.38) and who were black (OR = 4.70; 95%CI: 1.85; 11.95) were more likely to be obese. This study found an increase in the prevalence of obesity during the studied period, with increased odds for middle-aged and black people and decreased odds for those without a partner and those who practice physical activities in their leisure and as a form of commuting. These results can contribute to support programs and policies to control obesity.

O objetivo foi investigar a associação entre fatores sociodemográficos e comportamentais e a ocorrência da obesidade entre 2014 e 2021 em adultos da cidade de São Paulo, Brasil. Realizou-se estudo prospectivo com 1.241 adultos paulistanos, com 18 anos ou mais, participantes da coorte Inquérito de Saúde de São Paulo (ISA) - Atividade Física e Ambiente. O desfecho foi obesidade (sim/não), classificada por meio do índice de massa corporal e com pontos de corte específicos para cada faixa etária. As variáveis de exposição foram: sexo, idade, escolaridade, cor da pele, estado marital, coordenadoria regional de saúde, atividade física nos quatro domínios e comportamentos sedentários. Foram utilizados modelos de regressão logística multinível para a análise longitudinal. Houve aumento significativo de 27,7% na prevalência de obesidade (de 22,6% para 28,9%). Pessoas que praticavam ao menos 150 minutos semanais de atividade física no lazer (OR = 0,44; IC95%: 0,26; 0,76), entre 10 e 150 minutos semanais de atividade física de deslocamento (OR = 0,49; IC95: 0,30; 0,80) e sem companheiro(a) (OR = 0,47; IC95%: 0,28; 0,78) tiveram menos chances de ter obesidade. Pessoas entre 40 e 59 anos (OR = 5,00; IC95%: 2,02; 12,38) e de cor de pele preta (OR = 4,70; IC95%: 1,85; 11,95) apresentaram maiores chances de ter obesidade. O estudo identificou um aumento na prevalência de obesidade durante o período, com aumento nas chances para pessoas de meia idade e cor da pele preta, e diminuição nas chances para pessoas que vivem sem companheiro(a) e para praticantes de atividades físicas no lazer e como forma de deslocamento. Esses resultados podem contribuir para dar suporte a programas e políticas para o controle da obesidade.

El objetivo fue investigar la asociación entre factores sociodemográficos y conductuales y la presencia de la obesidad entre 2014 y 2021 en adultos de la ciudad de São Paulo, Brasil. Se realizó un estudio prospectivo con 1.241 adultos paulistanos, con 18 años o más, participantes de la cohorte Encuesta de Salud de São Paulo (ISA) - Actividad Física y Ambiente. El resultado fue obesidad (sí/no), clasificada a través del índice de masa corporal y con puntos de corte específicos para cada grupo de edad. Las variables de exposición fueron: sexo, edad, escolaridad, color de la piel, estado civil, coordinación regional de salud, actividad física en los cuatro dominios y comportamientos sedentarios. Para los análisis longitudinales se utilizaron modelos de regresión logística multinivel. Hubo un aumento significativo del 27,7% en la prevalencia de la obesidad (del 22,6% al 28,9%). Las personas que practicaron al menos 150 minutos semanales de actividad física por ocio (OR = 0,44; IC95%: 0,26; 0,76), entre 10 y 150 minutos semanales de actividad física de desplazamiento (OR = 0,49; IC95%: 0,30; 0,80) y sin compañero(a) (OR = 0,47; IC95%: 0,28; 0,78) tuvieron menos probabilidades de tener obesidad. Las personas entre 40 y 59 años (OR = 5,00; IC95%: 2,02; 12,38) y de color de piel negra (OR = 4,70; IC95%: 1,85; 11,95) presentaron mayores probabilidades de tener obesidad. El estudio identificó un aumento en la prevalencia de la obesidad durante ese período, con un aumento en las posibilidades para las personas de mediana edad y color de la piel negra, y una disminución en las posibilidades para las personas que viven sin pareja y para las que realizan actividades físicas por ocio y como forma de desplazamiento. Estos resultados pueden contribuir a apoyar los programas y políticas para el control de la obesidad.

An improved expression and purification protocol enables the structural characterization of Mnt1, an antifungal target from Candida albicans.

BACKGROUND: Candida albicans is one of the most prevalent fungi causing infections in the world. Mnt1 is a mannosyltransferase that participates in both the cell wall biogenesis and biofilm growth of C. albicans. While the cell wall performs crucial functions in pathogenesis, biofilm growth is correlated with sequestration of drugs by the extracellular matrix. Therefore, antifungals targeting CaMnt1 can compromise fungal development and potentially also render Candida susceptible to drug therapy. Despite its importance, CaMnt1 has not yet been purified to high standards and its biophysical properties are lacking. RESULTS: We describe a new protocol to obtain high yield of recombinant CaMnt1 in Komagataella phaffii using methanol induction. The purified protein's identity was confirmed by MALDI-TOF/TOF mass spectroscopy. The Far-UV circular dichroism (CD) spectra demonstrate that the secondary structure of CaMnt1 is compatible with a protein formed by α-helices and ß-sheets at pH 7.0. The fluorescence spectroscopy results show that the tertiary structure of CaMnt1 is pH-dependent, with a greater intensity of fluorescence emission at pH 7.0. Using our molecular modeling protocol, we depict for the first time the ternary complex of CaMnt1 bound to its two substrates, which has enabled the identification of residues involved in substrate specificity and catalytic reaction. Our results corroborate the hypothesis that Tyr209 stabilizes the formation of an oxocarbenium ion-like intermediate during nucleophilic attack of the acceptor sugar, opposing the double displacement mechanism proposed by other reports. CONCLUSIONS: The methodology presented here can substantially improve the yield of recombinant CaMnt1 expressed in flask-grown yeasts. In addition, the structural characterization of the fungal mannosyltransferase presents novelties that can be exploited for new antifungal drug's development.

Cancer evaluation in dogs using cerumen as a source for volatile biomarker prospection.

Cancer is one of the deadliest diseases in humans and dogs. Nevertheless, most tumor types spread faster in canines, and early cancer detection methods are necessary to enhance animal survival. Here, cerumen (earwax) was tested as a source of potential biomarkers for cancer evaluation in dogs. Earwax samples from dogs were collected from tumor-bearing and clinically healthy dogs, followed by Headspace/Gas Chromatography-Mass Spectrometry (HS/GC-MS) analyses and multivariate statistical workflow. An evolutionary-based multivariate algorithm selected 18 out of 128 volatile metabolites as a potential cancer biomarker panel in dogs. The candidate biomarkers showed a full discrimination pattern between tumor-bearing dogs and cancer-free canines with high accuracy in the test dataset: an accuracy of 95.0% (75.1-99.9), and sensitivity and specificity of 100.0% and 92.9%, respectively. In summary, this work raises a new perspective on cancer diagnosis in dogs, being carried out painlessly and non-invasive, facilitating sample collection and periodic application in a veterinary routine.

A veterinary cerumenomic assay for bovine laminitis identification.

Bovine laminitis disorder results in animal welfare and economic concerns in dairy and beef farms worldwide. However, the affected metabolic pathways, pathophysiologic characteristics, and inflammatory mechanisms remain unclear, hampering the development of new diagnostics. Using cerumen (earwax) as a source of volatile metabolites (cerumenomic) that carry valuable biological information has interesting implications for veterinary medicine. Nonetheless, up to now, no applications of veterinary cerumenomic assays have been made to identify bovine laminitis. This work aims to develop a veterinary cerumenomic assay for bovine laminitis identification that is non-invasive, robust, accurate, and sensitive to detecting the metabolic disturbances in bovine volatile metabolome. Twenty earwax samples (10 from healthy/control calves and 10 from laminitis calves) were collected from Nellore cattle, followed by Headspace/Gas Chromatography-Mass Spectrometry (HS/GC-MS) analysis and biomarker selection in two multivariate approaches: semiquantitative (intensity data) and semiqualitative (binary data). Following the analysis, cerumen volatile metabolites were indicated as candidate biomarkers for identifying bovine laminitis by monitoring their intensity or occurrence. In the semiquantitative strategy, the p-cresol presented the highest diagnostic figures of merit (area under the curve: 0.845, sensitivity: 0.700, and specificity: 0.900). Regarding the binary approach, a panel combining eight variables/volatiles, with formamide being the most prominent one, showed an area under the curve, sensitivity, and specificity of 0.97, 0.81, and 0.90, respectively. In summary, this work describes the first veterinary cerumenomic assay for bovine laminitis that indicates new metabolites altered during the inflammatory condition, paving the way for developing laminitis early diagnosis by monitoring the cerumen metabolites.

Crystal structure and interconversion of monomers and domain-swapped dimers of the walnut tree phytocystatin.

Biotechnological applications of phytocystatins have garnered significant interest due to their potential applications in crop protection and improve crop resistance to abiotic stress factors. Cof1 and Wal1 are phytocystatins derived from Coffea arabica and Juglans regia, respectively. These plants hold significant economic value due to coffee's global demand and the walnut tree's production of valuable timber and widely consumed walnuts with culinary and nutritional benefits. The study involved the heterologous expression in E. coli Lemo 21(DE3), purification by immobilized metal ion affinity and size exclusion chromatography, and biophysical characterization of both phytocystatins, focusing on isolating and interconverting their monomers and dimers. The crystal structure of the domain-swapped dimer of Wal1 was determined revealing two domain-swapped dimers in the asymmetric unit, an arrangement reminiscent of the human cystatin C structure. Alphafold models of monomers and Alphafold-Multimer models of domain-swapped dimers of Cof1 and Wal1 were analyzed in the context of the crystal structure. The methodology and data presented here contribute to a deeper understanding of the oligomerization mechanisms of phytocystatins and their potential biotechnological applications in agriculture.

Fatores sociodemográficos e comportamentais da obesidade: um estudo longitudinal / Obesity sociodemographic and behavioral factors: a longitudinal study / Factores sociodemográficos y conductuales de la obesidad: un estudio longitudinal

Resumo: O objetivo foi investigar a associação entre fatores sociodemográficos e comportamentais e a ocorrência da obesidade entre 2014 e 2021 em adultos da cidade de São Paulo, Brasil. Realizou-se estudo prospectivo com 1.241 adultos paulistanos, com 18 anos ou mais, participantes da coorte Inquérito de Saúde de São Paulo (ISA) - Atividade Física e Ambiente. O desfecho foi obesidade (sim/não), classificada por meio do índice de massa corporal e com pontos de corte específicos para cada faixa etária. As variáveis de exposição foram: sexo, idade, escolaridade, cor da pele, estado marital, coordenadoria regional de saúde, atividade física nos quatro domínios e comportamentos sedentários. Foram utilizados modelos de regressão logística multinível para a análise longitudinal. Houve aumento significativo de 27,7% na prevalência de obesidade (de 22,6% para 28,9%). Pessoas que praticavam ao menos 150 minutos semanais de atividade física no lazer (OR = 0,44; IC95%: 0,26; 0,76), entre 10 e 150 minutos semanais de atividade física de deslocamento (OR = 0,49; IC95: 0,30; 0,80) e sem companheiro(a) (OR = 0,47; IC95%: 0,28; 0,78) tiveram menos chances de ter obesidade. Pessoas entre 40 e 59 anos (OR = 5,00; IC95%: 2,02; 12,38) e de cor de pele preta (OR = 4,70; IC95%: 1,85; 11,95) apresentaram maiores chances de ter obesidade. O estudo identificou um aumento na prevalência de obesidade durante o período, com aumento nas chances para pessoas de meia idade e cor da pele preta, e diminuição nas chances para pessoas que vivem sem companheiro(a) e para praticantes de atividades físicas no lazer e como forma de deslocamento. Esses resultados podem contribuir para dar suporte a programas e políticas para o controle da obesidade.

Abstract: This study aimed to investigate the association between sociodemographic and behavioral factors and obesity from 2014 to 2021 in adults in São Paulo city, Brazil. A prospective study was carried out with 1,241 adults aged 18 years or above who lived in São Paulo and participated in the São Paulo Health Survey cohort (ISA) - Physical Activity and Environment. The outcome was obesity (yes/no), classified by body mass index and specific cut-off points for each age group. The exposure variables included sex, age, education, skin color, marital status, regional health coordination, physical activity in the four domains, and sedentary behaviors. Multilevel logistic regression models were used for longitudinal analysis. This study found a significant increase (27.7%) in the prevalence of obesity (from 22.6% to 28.9%). People who practiced at least 150 minutes of leisure-time physical activity per week (OR = 0.44; 95%CI: 0.26; 0.76), between 10 and 150 minutes per week of commuting physical activity (OR = 0.49; 95%CI: 0.30; 0.80), and had no partner (OR = 0.47; 95%CI: 0.28; 0.78) were less likely to be obese. People aged from 40 to 59 years (OR = 5.00; 95%CI: 2.02; 12.38) and who were black (OR = 4.70; 95%CI: 1.85; 11.95) were more likely to be obese. This study found an increase in the prevalence of obesity during the studied period, with increased odds for middle-aged and black people and decreased odds for those without a partner and those who practice physical activities in their leisure and as a form of commuting. These results can contribute to support programs and policies to control obesity.

Resumen: El objetivo fue investigar la asociación entre factores sociodemográficos y conductuales y la presencia de la obesidad entre 2014 y 2021 en adultos de la ciudad de São Paulo, Brasil. Se realizó un estudio prospectivo con 1.241 adultos paulistanos, con 18 años o más, participantes de la cohorte Encuesta de Salud de São Paulo (ISA) - Actividad Física y Ambiente. El resultado fue obesidad (sí/no), clasificada a través del índice de masa corporal y con puntos de corte específicos para cada grupo de edad. Las variables de exposición fueron: sexo, edad, escolaridad, color de la piel, estado civil, coordinación regional de salud, actividad física en los cuatro dominios y comportamientos sedentarios. Para los análisis longitudinales se utilizaron modelos de regresión logística multinivel. Hubo un aumento significativo del 27,7% en la prevalencia de la obesidad (del 22,6% al 28,9%). Las personas que practicaron al menos 150 minutos semanales de actividad física por ocio (OR = 0,44; IC95%: 0,26; 0,76), entre 10 y 150 minutos semanales de actividad física de desplazamiento (OR = 0,49; IC95%: 0,30; 0,80) y sin compañero(a) (OR = 0,47; IC95%: 0,28; 0,78) tuvieron menos probabilidades de tener obesidad. Las personas entre 40 y 59 años (OR = 5,00; IC95%: 2,02; 12,38) y de color de piel negra (OR = 4,70; IC95%: 1,85; 11,95) presentaron mayores probabilidades de tener obesidad. El estudio identificó un aumento en la prevalencia de la obesidad durante ese período, con un aumento en las posibilidades para las personas de mediana edad y color de la piel negra, y una disminución en las posibilidades para las personas que viven sin pareja y para las que realizan actividades físicas por ocio y como forma de desplazamiento. Estos resultados pueden contribuir a apoyar los programas y políticas para el control de la obesidad.

Spray drying of cellulases produced by Bacillus sp SMIA-2

Microbial cellulases present biotechnological potential to be used in several industries, including food, brewery and wine, industrial waste for chemical feedstock, animal feed, pulp and paper, agriculture, textile and detergent production. In this work, cellulases produced by the thermophilic Bacillus sp. SMIA-2 in submerged cultures containing sugarcane bagasse, corn steep liquor and passion fruit rind flour were spray-dried, in an attempt to improve their stability for industrial purposes. The cellulases were spray dried and analyzed before and after the drying process and subsequent storage. A Central Composite Design (CCD) 23 was used to investigate the effect of different concentrations of arabic gum and microcrystalline cellulose, as well as the spray dryer inlet temperature on the cellulase spray drying process. The results evidenced that the combination of 1.0 % (w/v) arabic gum and 1.0% (w/v) microcrystalline cellulose, at inlet temperature of 70 °C, was effective in maintaining the activities of both avicelases (avicel-hydrolyzing enzymes) and carboxymethylcellulases (carboxymethylcellulose-hydrolyzing enzymes - CMCase). The dried avicelase was completely preserved when stored at 5oC, while the CMCase retained 89% of its activity, which indicates promising potential for industrial uses, especially in detergent formulations.

Evaluation of the effects of the recommended oral dose of diflubenzuron on bovine sperm and oocyte quality using CASA and OPU-IVEP.

Introduction: The aim of this study was to evaluate potential effects of diflubenzuron on the production and quality of gametes, and on in vitro embryo production (IVEP) outcomes, in cattle. Methods: Two experiments were performed, the first to evaluate effects on semen, and the second on cumulus-oocyte complexes (COC) and on IVEP. Nelore (Bos taurus indicus) bulls (n = 14) or heifers (n = 16) were allocated into control (CG) or treatment (DIF) groups. All groups received a mineral mix supplement added (DIF) or not (CG) with diflubenzuron (30 mg/head/day), during 8 weeks. Animals were weighed and blood samples were collected throughout the experimental period. Every other week, bulls were subjected to semen collection and heifers to transvaginal ultrasound-guided follicle aspiration sessions. Semen underwent physical and morphological evaluation, and samples were stored for further computer-assisted sperm analysis. The COC recovered were evaluated according to morphology and those classified as viable were sent to an IVEP laboratory. Results: Diflubenzuron had no effect (P > 0.05) on average body weight or in any blood hematological or biochemical endpoints, regardless of gender. In experiment 1, there was no difference (P > 0.05) between DIF and CG groups for sperm concentration, morphology, or kinetics. In experiment 2, there was also no effect of diflubenzuron on the number of total, viable, or grade I oocytes, as well as on cleavage or blastocyst rates (P > 0.05). Discussion: In summary, the oral administration of diflubenzuron, within the recommended dose, has no short-term negative effects on sperm production and quality or on oocyte yield and developmental potential in vitro, in cattle.

Identification of novel Zika virus NS3 protease inhibitors with different inhibition modes by integrative experimental and computational approaches.

Zika virus (ZIKV) infection is associated with severe neurological disorders and congenital malformation. Despite efforts to eradicate the disease, there is still neither vaccine nor approved drugs to treat ZIKV infection. The NS2B-NS3 protease is a validated drug target since it is essential to polyprotein virus maturation. In the present study, we describe an experimental screening of 2,320 compounds from the chemical library of the Muséum National d'Histoire Naturelle of Paris on ZIKV NS2B-NS3 protease. A total of 96 hits were identified with 90% or more of inhibitory activity at 10 µM. Amongst the most active compounds, five were analyzed for their inhibitory mechanisms by kinetics assays and computational approaches such as molecular docking. 2-(3-methoxyphenoxy) benzoic acid (compound 945) show characteristics of a competitive inhibition (Ki = 0.49 µM) that was corroborated by its molecular docking at the active site of the NS2B-NS3 protease. Taxifolin (compound 2292) behaves as an allosteric inhibitor whereas 3,8,9-trihydroxy-2-methyl-1H-phenalen-1-one (compound 128), harmol (compound 368) and anthrapurpurin (compound 1499) show uncompetitive inhibitions. These new NS2B-NS3 protease inhibitors are valuable hits to further hit-to-lead optimization.

Purification and Biological Properties of Raniseptins-3 and -6, Two Antimicrobial Peptides from Boana raniceps (Cope, 1862) Skin Secretion.

The number of multidrug-resistant pathogenic microorganisms has been growing in recent years, most of which is due to the inappropriate use of the commercial antibiotics that are currently available. The dissemination of antimicrobial resistance represents a serious global public health problem. Thus, it is necessary to search for and develop new drugs that can act as antimicrobial agents. Antimicrobial peptides are a promising alternative for the development of new therapeutic drugs. Anurans' skin glands are a rich source of broad-spectrum antimicrobial compounds and hylids, a large and diverse family of tree frogs, are known as an important source of antimicrobial peptides. In the present study, two novel antimicrobial peptides, named Raniseptins-3 and -6, were isolated from Boana raniceps skin secretion and their structural and biological properties were evaluated. Raniseptins-3 and -6 are cationic, rich in hydrophobic residues, and adopt an α-helix conformation in the presence of SDS (35 mM). Both peptides are active against Gram-negative bacteria and Gram-positive pathogens, with low hemolytic activity at therapeutic concentrations. No activity was observed for yeasts, but the peptides are highly cytotoxic against B16F10 murine melanoma cells and NIH3T3 mouse fibroblast cells. None of the tested compounds showed improvement trends in the MTT and LDH parameters of MHV-3 infected cells at the concentrations tested.

Mobile application-based oximetry: a potential toolfor appropriate referral of patients with respiratory symptoms examined via telemedicine.

OBJECTIVE: To calculate the positive likelihood ratio to determine whether telemedicine is able to optimize referral to the emergency department. METHODS: Unicenter study with 182 consecutive patients admitted to Hospital Israelita Albert Einstein due to respiratory symptoms. All patients were submitted to oxygen saturation measurement using the standard method Welch Allyn finger device vital sign monitor and a 2-minute evaluation (Binah.ai mobile application). The reproducibility of oxygen saturation measurements made with both methods was investigated using interclass correlation coefficients and analysis of dispersion. Bland-Altman plots were constructed and kappa concordance coefficients used to examine data normality. Accuracy was also estimated. RESULTS: Oxygen saturation measurement differences between methods were ≤2% in more than 85% of cases. The mean difference (bias) between methods was near zero (0.835; Bland-Altman analysis). Oxygen saturation measurements made using the Binah.ai mobile application had an average ability to detect patients with altered oxygen saturation levels compared to the conventional method (ROC analysis). The positive likelihood ratio of the mobile application was 6.23. CONCLUSION: Mobile applications for oxygen saturation measurement are accessible user-friendly tools with moderate impact on clinical telemedicine evaluation of patients with respiratory symptoms, and may optimize referral to the emergency department.

Interaction models between peptide substrate and Alphavirus Protease nsP2 of Chikungunya and Mayaro and implications to the mechanism of action.

The Arbovirus (Arthropod-borne virus) is a group which comprises viruses whose transmission is carried out by arthropod vectors infecting vertebrates. Some arboviruses related to human diseases have been given considerable relevance as Chikungunya and Mayaro of the family Togaviridae, genus Alphavirus. The lack of proper specific treatment has prompted the requirement for deeper structural studies that could unveil leads to new drugs. Among possible targets, viral proteases are recognized as proteins with big potential. These proteins, termed nsP2 in Alphavirus, have the function of cleaving certain regions of the viral polyprotein, being vital to the viral cycle. In this research, we used docking and molecular dynamics to analyze the contact between the protease nsP2 of Alphavirus Chikungunya and Mayaro and substrates formed by peptides with ten amino acid residues. A model of the Mayaro nsP2 was constructed based on homologous proteases. Our study suggests that the glycine specificity motif, a region where a highly conserved glycine residue in position P2 of the protease substrate is positioned, facilitates the nucleophilic attack by assisting in placing the P1 carbonyl group carbon. Stabilization of different substrate regions maybe explained by relevant contacts with the enzyme. Besides that, the phi and psi angles in the outlier region of the Ramachandran plot found for the P2 glycine of the Chikungunya substrate seems to indicate the necessity of this residue that can accommodate angles not allowed to other residues.Communicated by Ramaswamy H. Sarma.

Multiple facets of biodiversity are threatened by mining-induced land-use change in the Brazilian Amazon

Aim Mining is increasingly pressuring areas of critical importance for biodiversity conservation, such as the Brazilian Amazon. Biodiversity data are limited in the tropics, restricting the scope for risks to be appropriately estimated before mineral licensing decisions are made. As the distributions and range sizes of other taxa differ markedly from those of vertebrates—the common proxy for analysis of risk to biodiversity from mining—whether mining threatens lesser-studied taxonomic groups differentially at a regional scale is unclear. Location Brazilian Amazon. Methods We assess risks to several facets of biodiversity from industrial mining by comparing mining areas (within 70 km of an active mining lease) and areas unaffected by mining, employing species richness, species endemism, phylogenetic diversity and phylogenetic endemism metrics calculated for angiosperms, arthropods and vertebrates. Results Mining areas contained higher densities of species occurrence records than the unaffected landscape, and we accounted for this sampling bias in our analyses. None of the four biodiversity metrics differed between mining and nonmining areas for vertebrates. For arthropods, species endemism was greater in mined areas. Mined areas also had greater angiosperm species richness, phylogenetic diversity and phylogenetic endemism, although less species endemism than unmined areas. Main Conclusions Unlike for vertebrates, facets of angiosperm and arthropod diversity are relatively higher in areas of mining activity, underscoring the need to consider multiple taxonomic groups and biodiversity facets when assessing risk and evaluating management options for mining threats. Particularly concerning is the proximity of mining to areas supporting deep evolutionary history, which may be impossible to recover or replace. As pressures to expand mining in the Amazon grow, impact assessments with broader taxonomic reach and metric focus will be vital to conserving biodiversity in mining regions.

Mobile application-based oximetry: a potential toolfor appropriate referral of patients with respiratory symptoms examined via telemedicine

ABSTRACT Objective To calculate the positive likelihood ratio to determine whether telemedicine is able to optimize referral to the emergency department. Methods Unicenter study with 182 consecutive patients admitted to Hospital Israelita Albert Einstein due to respiratory symptoms. All patients were submitted to oxygen saturation measurement using the standard method Welch Allyn finger device vital sign monitor and a 2-minute evaluation (Binah.ai mobile application). The reproducibility of oxygen saturation measurements made with both methods was investigated using interclass correlation coefficients and analysis of dispersion. Bland-Altman plots were constructed and kappa concordance coefficients used to examine data normality. Accuracy was also estimated. Results Oxygen saturation measurement differences between methods were ≤2% in more than 85% of cases. The mean difference (bias) between methods was near zero (0.835; Bland-Altman analysis). Oxygen saturation measurements made using the Binah.ai mobile application had an average ability to detect patients with altered oxygen saturation levels compared to the conventional method (ROC analysis). The positive likelihood ratio of the mobile application was 6.23. Conclusion Mobile applications for oxygen saturation measurement are accessible user-friendly tools with moderate impact on clinical telemedicine evaluation of patients with respiratory symptoms, and may optimize referral to the emergency department.

Radical prostatectomies for treatment of prostate cancer: trends in a ten-year period in public health services in the city of São Paulo, Brazil.

OBJECTIVE: To analyze the characteristics of public health services related to radical prostatectomy, according to hospital volume of surgeries and stratified as academic and non-academic centers. METHODS: An ecological study was conducted using a database available in TabNet platform of the Unified Health System Department of Informatics. Number of surgeries, length of hospital stay, length of stay in intensive care unit, in-hospital mortality rate, and cost of hospitalization were evaluated. The hospitals were divided into three subgroups according to surgery volume (tercile), and results were compared. The same comparisons were made among academic and non-academic centers. We considered academic centers those providing Urology residency program. RESULTS: A total of 11,259 radical prostatectomies were performed in the city of São Paulo between 2008 and 2018. We observed a significant trend of increase in radical prostatectomies for treating prostate cancer over the years (p=0.007). The length of stay in intensive care unit, and number of deaths were not statistically different among centers with diverse surgery volume, nor between academic and non-academic centers. However, length of hospital stay was significantly shorter in academic centers (p=0.043), while cost of hospitalization was significantly higher in high-volume center compared to low- (p<0.001) and intermediate-volume centers (p<0.001). CONCLUSION: Length of hospital stay for radical prostatectomies performed in public services in the city of São Paulo was shorter in academic centers, whereas hospitals with a high volume of surgeries showed greater cost of hospitalization.

Prognosis of Extended-Spectrum-Beta-Lactamase-Producing Agents in Emphysematous Pyelonephritis-Results from a Large, Multicenter Series.

BACKGROUND: Emphysematous pyelonephritis (EPN) is a necrotizing infection of the kidney and surrounding tissues with significant mortality. We aimed to assess the clinical factors and their influence on prognosis in patients being managed for EPN with and without ESBL-producing bacteria and to identify if those with EPN due to ESBL infections fared any different. METHODS: A retrospective analysis was performed on patients with EPN diagnosis from 22 centers across 11 countries (between 2013 and 2020). Demographics, clinical presentation, biochemical parameters, radiological features, microbiological characteristics, and therapeutic management were assessed. Univariable and multivariable analyses were performed to determine the independent variables associated with ESBL pathogens. A comparison of ESBL and non-ESBL mortality was performed evaluating treatment modality. RESULTS: A total of 570 patients were included. Median (IQR) age was 57 (47-65) years. Among urine cultures, the most common isolated pathogen was Escherichia coli (62.2%). ESBL-producing agents were present in 291/556 urine cultures (52.3%). In multivariable analysis, thrombocytopenia (OR 1.616 95% CI 1.081-2.413, p = 0.019), and Huang-Tseng type 4 (OR 1.948 95% CI 1.005-3.778, p= 0.048) were independent predictors of ESBL pathogens. Patients with Huang-Tseng Scale type 1 had 55% less chance of having ESBL-producing pathogens (OR 1.616 95% CI 1.081-2.413, p = 0.019). Early nephrectomy (OR 2.3, p = 0.029) and delayed nephrectomy (OR 2.4, p = 0.015) were associated with increased mortality in patients with ESBL infections. Conservative/minimally invasive management reported an inverse association with mortality (OR 0.314, p = 0.001). CONCLUSIONS: ESBL bacteria in EPN were not significantly associated with mortality in EPN. However, ESBL infections were associated with poor prognosis when patients underwent nephrectomy compared conservative/minimally invasive management.

An Assessment of Mortality among Elderly Brazilians from Alcohol Abuse Diseases: A Longitudinal Study from 1996 to 2019.

Alcohol use disorder (AUD) is a worldwide public health problem, being an important aggravating factor of comorbidities found in the elderly, with the potential to increase mortality indicators for this age group. OBJECTIVE: To analyze alcohol-induced deaths in elderly people with alcohol-related disorder in Brazil between 1996 and 2019. METHODS: An ecological study was conducted with secondary data obtained from the Brazilian Unified Health System (SIM) Mortality Information System from 1996 to 2019. TabNet/DATASUS, Excel® 2016 and SPSS 21® were used to prepare the results. RESULTS: Between 1996 and 2019, 85,928 alcohol-induced deaths were recorded among the elderly (>60 years); in 1996, the lowest number of deaths was recorded (n = 1396), and in 2018, there were the highest number of deaths (n = 5667). In the profile of the elderly, there was a predominance of men (88%). Mortality from AUD was due to alcoholic liver disease (62.2%), followed by mental disorders due to alcohol use (37.3%). CONCLUSIONS: Coping with AUD is a public health problem that aims to reduce the number of deaths from diseases, conditions and injuries in which alcohol consumption is the causative agent, in addition to preventing deaths to which alcohol contributes.