Elevated NS1 serum levels reduce CD119 expression and CXCL-10 synthesis in patients with dengue hemorrhagic fever.

BACKGROUND: The intensity of dengue virus (DV) replication and circulating non-structural protein 1 (NS1) levels may promote changes in the human immune response and favor severe forms of infection. We investigated the correlations between NS1 with CXCL-8, CXCL-10, IFN-γ, and IL-12p40 serum levels, and IFN-γ receptor α chain (CD119) expression, and CXCL10 production by peripheral blood mononuclear cells (PBMCs) stimulated with recombinant IFN-γ in DV-infected patients with different clinical forms. METHODS: Dengue virus NS1, CXCL-8, CXCL-10, IFN-γ, and IL-12p40 serum levels were measured in 152 DV-infected patients with different clinical forms and 20 non-infected individuals (NI) using enzyme-linked immunosorbent assay (ELISA). In addition, we investigated the CXCL-10 production after in vitro IFN-γ stimulation of PBMCs from 48 DV-infected individuals (with different clinical forms of dengue fever) and 20 NI individuals using ELISA, and CD119 expression on CD14+ cells with flow cytometry. RESULTS: Patients with dengue hemorrhagic fever (DHF) had significantly higher NS1, CXCL-8, and CXCL-10 serum levels than those with classic dengue fever (DF). The response of PBMCs to IFN-γ stimulation was lower in patients with DHF than in those with DF or dengue with complications (DWC), with lower CD119 expression and reduced CXCL-10 synthesis. In addition, these alterations are associated with high NS1 serum levels. CONCLUSIONS: Patients with DHF reported high NS1 levels, low CD119 expression, and low CXCL-10 synthesis in PBMCs, which may be associated with infection progression and severity.

Can changes in land use in a semi-arid region of Brazil cause seasonal variation in energy partitioning and evapotranspiration?

Changes to forests due to deforestation, or their replacement by agricultural areas, alter evapotranspiration and the partitioning of available energy. This study investigated seasonal variations in the energy balance and evapotranspiration in landscapes under different levels of anthropogenic intervention in the semi-arid region of Brazil. Micrometeorological data was obtained from September 2020 to October 2022 for three areas of the semi-arid region: preserved Caatinga (CAA, native vegetation), Caatinga under regeneration (REGE) and a deforested area (DEFA). Here, we use the Bowen ratio energy balance method. Measurements were taken of global solar radiation, air temperature, relative humidity, vapour pressure deficit, rainfall, net radiation, latent heat flux, sensible heat flux, soil heat flux, evapotranspiration, volumetric soil water content and Normalised Difference Vegetation Index. Sensible heat flux was the dominant flux in both areas with 66% for preserved Caatinga vegetation, 63% for Caatinga under regeneration and 62% deforested area. The latent heat flux was equivalent to 28% of the net radiation for preserved Caatinga vegetation, Caatinga under regeneration and deforested area. The evapotranspiration in turn responded as a function of water availability, being higher during the rainy seasons, with average values of 1.82 mm day-1 for preserved Caatinga vegetation, 2.26 mm day-1 for Caatinga under regeneration and 1.25 mm day-1 for deforested area. The Bowen ratio presented values > 1 in deforested area, preserved Caatinga vegetation and Caatinga under regeneration. Thus, it can be concluded that the change in land use alters the energy balance components, promoting reductions in available energy and latent and sensible heat fluxes during the rainy-dry transition in the deforested area. In addition, the seasonality of energy fluxes depends on water availability in the environment.

Estimated quantification of residual volume in vaccines supplies and its impact on the Brazilian health system.

Between 2017 and 2021, the Brazilian Unified Health System (BUHS) administered a total of 527,903,302 doses of immunizations. Each immunization results in the presence of a residual volume (RV) due to syringe dead space (DS). The International Organization for Standardization 7886-1 allows a DS of up to 0.07mL in sterile single-use hypodermic syringes with volumes less than 5mL. This study aims to quantify the DS of immunization devices used in Brazil, study the best combinations of needles and syringes to minimize RV, estimate the number of wasted doses from 2017 to 2021, and evaluate the impact on the BUHS. Pneumococcal 10 vaccine with a 25x6mm needle and a regular 1mL syringe exhibited a significantly higher average RV (0.0826mL) and waste rate (14.42%). It was observed that for some intramuscular vaccines, there is less waste when using a 20x5.5mm needle compared to a 25x6mm needle. The use of syringes with plunger stoppers that penetrate the syringe barrel, denoted as low dead space syringes, results in less RV and an estimated difference in the waste rate of approximately 10% compared to the regular syringe. The estimated number of wasted doses from 2017 to 2021 by BUHS is approximately 32 million doses.

Elevated NS1 serum levels reduce CD119 expression and CXCL-10 synthesis in patients with dengue hemorrhagic fever

ABSTRACT Background: The intensity of dengue virus (DV) replication and circulating non-structural protein 1 (NS1) levels may promote changes in the human immune response and favor severe forms of infection. We investigated the correlations between NS1 with CXCL-8, CXCL-10, IFN-γ, and IL-12p40 serum levels, and IFN-γ receptor α chain (CD119) expression, and CXCL10 production by peripheral blood mononuclear cells (PBMCs) stimulated with recombinant IFN-γ in DV-infected patients with different clinical forms. Methods: Dengue virus NS1, CXCL-8, CXCL-10, IFN-γ, and IL-12p40 serum levels were measured in 152 DV-infected patients with different clinical forms and 20 non-infected individuals (NI) using enzyme-linked immunosorbent assay (ELISA). In addition, we investigated the CXCL-10 production after in vitro IFN-γ stimulation of PBMCs from 48 DV-infected individuals (with different clinical forms of dengue fever) and 20 NI individuals using ELISA, and CD119 expression on CD14+ cells with flow cytometry. Results: Patients with dengue hemorrhagic fever (DHF) had significantly higher NS1, CXCL-8, and CXCL-10 serum levels than those with classic dengue fever (DF). The response of PBMCs to IFN-γ stimulation was lower in patients with DHF than in those with DF or dengue with complications (DWC), with lower CD119 expression and reduced CXCL-10 synthesis. In addition, these alterations are associated with high NS1 serum levels. Conclusions: Patients with DHF reported high NS1 levels, low CD119 expression, and low CXCL-10 synthesis in PBMCs, which may be associated with infection progression and severity.

Vaginal temperature modeling in Holstein cows and cluster analysis by the cophenetic correlation coefficient.

The objective was to establish a model for the prediction and characterization of vaginal temperature in Holstein cows, based on environmental predictors and thermal comfort indices, through cluster analysis, validation by the cophenetic correlation coefficient, and multiple regression analysis. The micrometeorological characterization of the site was carried out by recording the air temperature (Tair), the relative humidity (RH), the black globe temperature (BGT), the black globe temperature and humidity (BGHI), and dew point temperature (TDP). The recording of vaginal temperature (Tv) was performed in eight dairy cows using temperature sensors, equipped with data loggers, coupled with intravaginal devices. The data were analyzed using descriptive statistics and cluster analysis (CA) by using the hierarchical agglomerative method based on the value of the cophenetic correlation coefficient (CCC >0.70), in which representative physiological models were established, characterizing the Tv through multiple regression. In the afternoon the coefficient of variation (CV) was low for all variables, indicating homogeneity of the meteorological variables and efficiency of the ventilation system. The temperature and humidity index (THI) was mild only on the morning. There was a variation of 0.28 °C of Tv between shifts, sufficient to characterize the condition of comfort and stress of the animal, with values above 39 °C indicating animal stress. Tv showed strong correlation with BGT, Tair, TDP and RH, assuming that physiological variables, such as Tv, tend to have greater relationship with abiotic variables. Empirical models were established for estimating Tv based on the analyses performed in this study. Model 1 is recommended for TDP ranges of 14.00-21.00 °C and RH of 30-100%, while model 2 can be used for Tair situations up to 35 °C. The regression models for estimating Tv are promising for characterizing the thermal comfort of dairy cows housed in compost barn systems.

Cytotoxic and Antileishmanial Effects of the Monoterpene ß-Ocimene.

Leishmaniasis is a group of infectious-parasitic diseases with high mortality rates, and endemic in many regions of the globe. The currently available drugs present serious problems such as high toxicity, costs, and the emergence of drug resistance. This has stimulated research into new antileishmania drugs based on natural products and their derivatives. ß-Ocimene is a monoterpene found naturally in the essential oils of many plant species which presents antileishmanial activity, and which has not yet been evaluated for its potential to inhibit the etiological agent of leishmaniasis. The aim of this work was to evaluate the activity of ß-ocimene against Leishmania amazonensis, its cytotoxicity, and potential mechanisms of action. ß-Ocimene presented direct activity against the parasite, with excellent growth inhibition of promastigotes (IC50 = 2.78 µM) and axenic amastigotes (EC50 = 1.12 µM) at concentrations non-toxic to RAW 264.7 macrophages (CC50 = 114.5 µM). The effect is related to changes in membrane permeability and resulting abnormalities in the parasitic cell shape. These were, respectively, observed in membrane integrity and atomic force microscopy assays. ß-Ocimene was also shown to act indirectly, with greater activity against intra-macrophagic amastigotes (EC50 = 0.89 µM), increasing TNF-α, nitric oxide (NO), and reactive oxygen species (ROS), with lysosomal effects, as well as promoting decreases in IL-10 and IL-6. Against intra-macrophagic amastigote forms the selectivity index was higher than the reference drugs, being 469.52 times more selective than meglumine antimoniate, and 42.88 times more selective than amphotericin B. Our results suggest that ß-ocimene possesses promising in vitro antileishmania activity and is a potential candidate for investigation in in vivo assays.

Inflammatory Response and Activation of Coagulation after COVID-19 Infection.

SARS-CoV-2 (COVID-19) infection is responsible for causing a disease with a wide spectrum of clinical presentations. Predisposition to thromboembolic disease due to excessive inflammation is also attributed to the disease. The objective of this study was to characterize the clinical and laboratory aspects of hospitalized patients, in addition to studying the pattern of serum cytokines, and associate them with the occurrence of thromboembolic events. METHODOLOGY: A retrospective cohort study with 97 COVID-19 patients hospitalized from April to August 2020 in the Triângulo Mineiro macro-region was carried out. A review of medical records was conducted to evaluate the clinical and laboratory aspects and the frequency of thrombosis, as well as the measurement of cytokines, in the groups that presented or did not present a thrombotic event. RESULTS: There were seven confirmed cases of thrombotic occurrence in the cohort. A reduction in the time of prothrombin activity was observed in the group with thrombosis. Further, 27.8% of all patients had thrombocytopenia. In the group that had thrombotic events, the levels of IL1b, IL-10, and IL2 were higher (p < 0.05). CONCLUSIONS: In the studied sample, there was an increase in the inflammatory response in patients with thrombotic events, confirmed by the increase in cytokines. Furthermore, in this cohort, a link was observed between the IL-10 percentage and an increased chance of a thrombotic event.

Exploring the functionalization of Ti-6Al-4V alloy with the novel antimicrobial peptide JIChis-2 via plasma polymerization.

This study aimed to characterize the immobilization of the novel JIChis-2 peptide on the Ti-6Al-4V alloy, widely used in the biomedical sector. The antimicrobial activity of JIChis-2 was evaluated in the Gram-negative bacterium E. coli. Its immobilization occurred by inducing the formation of covalent bonds between the N-terminus of the peptides and the surface previously submitted to acrylic acid polymerization via the PECVD technique. Coated and uncoated surfaces were characterized by FTIR, AFM, SEM and EDX. Studies of global and localized corrosion were carried out, seeking to explore the effects triggered by surface treatment in an aggressive environment. Additionally, the ability of the functionalized material to prevent E. coli biofilm formation evidenced that the strategy to immobilize JIChis-2 in the Ti-6Al-4V alloy via PECVD of acrylic acid resulted in the development of a functional material with antibiofilm properties.

Factors for unfavorable evolution of snakebites in children

Pediatric patients have particularities in the clinical manifestations and complications of snakebite envenomation (SBEs), and few studies have examined this population. The objective of this paper was to study snakebites in a pediatric age group treated at a reference hospital and to evaluate factors associated with unfavorable evolution. A cross-sectional study with a clinical-epidemiological description and identification of the factors related to unfavorable evolution in patients aged <19 years old seen from January, 2018 to November, 2019 was performed. Complications related to the SBE, such as compartment syndrome, secondary infection, extensive necrosis, hemorrhage, and kidney damage, were considered unfavorable evolution. From the 325 patients in the sample, 58 were aged <19 years old; 40 (69%), 0-12 years old; and 18 (31%), 13-18 years old. All patients had local manifestations (mild, moderate, and severe), and 36 (62%) had an unfavorable evolution. Fourteen (24.1%) patients had compartment syndrome, with a significant risk association between 0 and 12 years old (p = 0.019). Two factors significantly contributed to unfavorable evolution: the timing from the bite to medical care being ≥6 hours and additional antivenom therapy needed. We conclude that the younger the patient, the smaller body segment affected, leading to disproportionality between the affected area and the amount of inoculated venom, contribute to more frequent local manifestations and complications in children rather than adults. Because of the relationship between body area and vascular volume in children differs from that in adults, the same volume of venom inoculated by snakes will be disproportionate in these two groups. Therefore, in the treatment of pediatric patients, increasing the volume of antivenom therapy is possibly necessary. Furthermore, as in adults, six hours between the bite to medical care increases the risk of complications and mortality.

Staphylococcus aureus and Enterococcus faecium isolated from pigeon droppings (Columba livia) in the external environment close to hospitals.

BACKGROUND: Domestic pigeons carry pathogens in their droppings, posing a potential public health problem. METHODS: The phenotypic and genotypic antimicrobial resistances of Staphylococcus aureus and Enterococcus faecium in the feces of urban pigeons near hospitals with intensive care units were measured. RESULTS: Twenty-nine samples showed Enterococcus growth, whereas one was positive for S. aureus. The S. aureus isolate was sensitive to the antibiotics tested via antibiogram, however resistance genes were identified. E. faecium isolates showed phenotypic resistance to gentamicin, erythromycin, and ciprofloxacin. CONCLUSIONS: Antimicrobial profiles harmful to health were demonstrated in bacterial pathogens isolated from the external environment of hospitals.

Association of recurrent upper respiratory tract infections with low production of oxygen intermediates in children

Abstract Objectives: Upper respiratory tract infections in children generally have significant morbidity and mortality. There is little data available about functional immaturity of the immune system and the child's susceptibility to infections at the beginning of their lives, thus, justifying a more specific immunological analysis. Method: Analysis of hemograms and innate and adaptive immune responses in 95 children between age 1 to 6 years with episodes of recurrent respiratory infections (test group n = 39) and without these episodes (control group n = 56) was carried out. The production of reactive oxygen intermediates by peripheral blood cells stimulated by phorbol myristate acetate was analyzed. Additionally, the number of B lymphocytes, auxiliary T lymphocytes, and cytotoxic cells was determined using flow cytometry. Results: Results from both groups did not show statistically significant differences in red blood cells, total leukocytes count, and the differential neutrophils, eosinophils, basophils, lymphocytes, and monocytes count. The analysis of the number of B lymphocytes, auxiliary T lymphocytes (LTCD4), and cytotoxic cells (LTCD8) also did not show any difference between both groups. However, the production of radical oxygen intermediates was significantly reduced in the test group as compared to the control group (p < 0.05). Conclusions: There was no difference in the analysis of hemograms, leukograms, or the number of lymphocytes, LTCD4, LTCD8, or LTCD19. The reduced production of oxygen intermediates in the affected group suggests that these children's microbicide capacity is compromised, which may be related to their recurrent respiratory infections.

Pulmonary hemorrhage in dengue: differential diagnosis with acute viral respiratory syndromes including COVID-19.

Clinical similarities among viral diseases become even more relevant considering the current scenario, especially in Brazil, where there is a high incidence of these diseases and overlapping seasonality. We report the case of a patient with acute clinical manifestations composed of predominant respiratory symptoms and alveolar hemorrhage in which three etiologies (dengue, influenza and COVID-19) were investigated concomitantly. Only the diagnosis of dengue was confirmed. Then, the patient's immunological profile in response to stimulation of mononuclear cells with dengue virus antigen was analyzed in an attempt to identify specific characteristics that could be associated with the clinical manifestation.

Staphylococcus aureus and Enterococcus faecium isolated from pigeon droppings (Columba livia) in the external environment close to hospitals

ABSTRACT Background: Domestic pigeons carry pathogens in their droppings, posing a potential public health problem. Methods: The phenotypic and genotypic antimicrobial resistances of Staphylococcus aureus and Enterococcus faecium in the feces of urban pigeons near hospitals with intensive care units were measured. Results: Twenty-nine samples showed Enterococcus growth, whereas one was positive for S. aureus. The S. aureus isolate was sensitive to the antibiotics tested via antibiogram, however resistance genes were identified. E. faecium isolates showed phenotypic resistance to gentamicin, erythromycin, and ciprofloxacin. Conclusions: Antimicrobial profiles harmful to health were demonstrated in bacterial pathogens isolated from the external environment of hospitals.

Hemolymph of triatomines presents fungistatic activity against Cryptococcus neoformans and improves macrophage function through MCP-I/TNF-α increase

Triatomines are blood-feeding arthropods belonging to the subfamily Triatominae (Hemiptera; Reduviidae), capable of producing immunomodulatory and water-soluble molecules in their hemolymph, such as antimicrobial peptides (AMPs). In this work, we evaluated the antifungal and immunomodulatory activity of the hemolymph of Meccus pallidipennis (MPH) and Rhodnius prolixus (RPH) against Cryptococcus neoformans. Methods: We assessed the activity of the hemolymph of both insects on fungal growth by a minimum inhibitory concentration (MIC) assay. Further, RAW 264.7 macrophages were cultivated with hemolymph and challenged with C. neoformans. Then, their phagocytic and killing activities were assessed. The cytokines MCP-1, IFN-γ, TNF-α, IL-10, IL-12, and IL-6 were measured in culture supernatants 4- and 48-hours post-infection. Results: Both hemolymph samples directly affected the growth rate of the fungus in a dose-dependent manner. Either MPH or RPH was capable of inhibiting fungal growth by at least 70%, using the lowest dilution (1:20). Treatment of RAW 264.7 macrophages with hemolymph of both insects was capable of increasing the production of MCP-I and TNF-α. In addition, when these cells were stimulated with hemolymph in the presence of C. neoformans, a 2- and a 4-fold increase in phagocytic rate was observed with MPH and RPH, respectively, when compared to untreated cells. For the macrophage killing activity, MPH decreased in approximately 30% the number of viable yeasts inside the cells compared to untreated control; however, treatment with RPH could not reduce the total number of viable yeasts. MPH was also capable of increasing MHC-II expression on macrophages. Regarding the cytokine production, MCP-I and TNF-α, were increased in the supernatant of macrophages treated with both hemolymphs, 4 and 48 hours after stimulation. Conclusion: These results suggested that hemolymph of triatomines may represent a source of molecules capable of presenting antifungal and immunomodulatory activity in macrophages during fungal infection.(AU)

Pulmonary hemorrhage in dengue: differential diagnosis with acute viral respiratory syndromes including COVID-19

ABSTRACT Clinical similarities among viral diseases become even more relevant considering the current scenario, especially in Brazil, where there is a high incidence of these diseases and overlapping seasonality. We report the case of a patient with acute clinical manifestations composed of predominant respiratory symptoms and alveolar hemorrhage in which three etiologies (dengue, influenza and COVID-19) were investigated concomitantly. Only the diagnosis of dengue was confirmed. Then, the patient's immunological profile in response to stimulation of mononuclear cells with dengue virus antigen was analyzed in an attempt to identify specific characteristics that could be associated with the clinical manifestation.

Reverse vaccinology and subtractive genomics approaches for identifying common therapeutics against Mycobacterium leprae and Mycobacterium lepromatosis

Mycobacterium leprae and Mycobacterium lepromatosis are gram-positive bacterial pathogens and the causative agents of leprosy in humans across the world. The elimination of leprosy cannot be achieved by multidrug therapy alone, and highlights the need for new tools and drugs to prevent the emergence of new resistant strains. Methods In this study, our contribution includes the prediction of vaccine targets and new putative drugs against leprosy, using reverse vaccinology and subtractive genomics. Six strains of Mycobacterium leprae and Mycobacterium lepromatosis (4 and 2 strains, respectively) were used for comparison taking Mycobacterium leprae strain TN as the reference genome. Briefly, we used a combined reverse vaccinology and subtractive genomics approach. Results As a result, we identified 12 common putative antigenic proteins as vaccine targets and three common drug targets against Mycobacterium leprae and Mycobacterium lepromatosis. Furthermore, the docking analysis using 28 natural compounds with three drug targets was done. Conclusions The bis-naphthoquinone compound Diospyrin (CID 308140) obtained from indigenous plant Diospyros spp. showed the most favored binding affinity against predicted drug targets, which can be a candidate therapeutic target in the future against leprosy.(AU)

Metabolic and immunological evaluation of patients with indeterminate and cardiac forms of Chagas disease.

ABSTRACT: Chagas disease affects approximately 7 million people, causing disability and mortality in the most productive life stages of infected individuals. Considering the lifestyle of the world population, metabolic syndrome is a synergistic factor for an increased cardiovascular risk of patients with Chagas disease.This study transversally evaluated the metabolic and immunological profiles of patients with indeterminate (IF) and cardiac (CF) forms of Chagas disease and their correlations with left ventricular dysfunction (LVD).Clinical and electrical bioimpedance analysis, levels of cytokines (interferon [IFN]-γ, tumor necrosis factor [TNF]-α, interleukin [IL]-17, IL-10, and IL-33) and adipocytokines (adiponectin, leptin, and resistin), metabolic syndrome components, and brain natriuretic peptide (BNP) levels were assessed in 57 patients (13 IF and 44 CF) with a mean age of 61.63 ±â€Š12.1 years. Chest x-ray, electrocardiogram, and echocardiogram were performed to classify the clinical forms.The CF group had a higher number of individuals with metabolic syndrome components blood pressure altered, while more participants in the CF group with LVD had low high-density lipoprotein (HDL) levels. The IF group had more participants with a higher waist-to-hip ratio (WHR). No significant difference was observed between metabolic syndrome, cytokine and adipocytokine level, and clinical forms of the disease or in relation to LVD.Individuals with the IF showed metabolic and immunological profiles compatible with increased disease control, whereas those with CF showed marked inflammatory immune response.

Spatial variability and exploratory inference of abiotic factors in barn compost confinement for cattle in the semiarid.

The assessment of environmental conditions in the creation of dairy cattle in feedlots must be carried out to identify possible causes of stress and assist producers in the decision-making process. The objective was to characterize the abiotic factors and bed temperature, in the comfort conditions of dairy cows, through geostatistics and exploratory data inference, in a Compost Barn System in the Brazilian semiarid region. The data were obtained in a milk production unit, located in the municipality of Lajedo, Pernambuco, Brazil. The variables air temperature (Tair), relative humidity (RH), wind speed (WS), illuminance (Lux), skin temperature (Ts), bed temperature (Tb) were recorded and the temperature index was determined and humidity (THI). Data were recorded at 9:00 a.m., 12:00 p.m., and 03:00 p.m., over 5 days in the summer season. For geostatistical analysis, the classic semivariances were determined. The principal component analysis was performed to establish an index that characterized the condition of animal comfort. The variables Tair, RH, Tb, and THI showed a low coefficient of variation for all times. The best fit to the models of the semivariograms was the Gaussian at 9:00 a.m. and 03:00 p.m., and the spherical at 12:00 p.m. The Tb spatial variability was low for all studied hours. Tair showed a strong correlation with Tb, due to the process of heat transfer by convection from the floor to the environment. Geostatistics and exploratory data analysis allowed the establishment of a comfort index for Compost Barn production systems in the Brazilian semiarid region (R2 = 0.996; p < 0.0001).