Fire effects on Atlantic Forest sites from a composition, structure and functional perspective.

Recently, some portions of the Atlantic Forest biome have been suffering an increase in forest fires, possibly changing its vegetation cover, composition, structure and functioning. Understanding these changes is critical to evaluate the present and future response of tropical forests to fire. Thus, the purpose of our study was to evaluate how diversity, structure and functioning of tree communities differed between burned and unburned sites. Two unburned and two burned forest patches were selected for floristic and phytosociological surveys. Then, we calculated species richness, Shannon diversity index, tree density and basal area, Importance Value Index for trees in each site and we assessed community weighted mean of six functional traits (maximum tree height, wood density, leaf length, leaf deciduousness, shade tolerance and dispersal mode). Diversity, species richness, tree density and basal area were similar between sites. We found changes in floristic composition, but did not verified variations in functional traits. Results indicate that recovery may be fast and that pioneer and early secondary species are occupying post burned sites (nine years old). One-time anthropogenic, superficial and low intensity fires might disrupt advanced stages of succession and start again the dynamics of species substitution.

Cylindrospermopsin impairs zebrafish (Danio rerio) embryo development.

Cyanotoxins are among common contaminants that can impair human, animal, and environmental health. Cylindrospermopsin (CYN) is an abundant form of cyanotoxins elevated following algal bloom in the water worldwide. Previous studies have described CYN effects on several organs in mammals. However, little is known about its toxicity mechanisms in other vertebrates. This study aims to characterize the developmental effects of CYN using zebrafish larvae as an aquatic model organism. A wide range of CYN concentrations (0-2000 µg/L) was tested using a morphometric approach for survival, hatching, various growth and developmental abnormalities. We also investigated the expression of genes related to oxidative stress, osmoregulation, and thyroid function. Exposure to CYN resulted in decreased growth, increased developmental abnormalities such as pericardial and yolk sac edema as well as swim bladder absence. In addition, CYN increased tr1a, and decreased dio1 and dio3 transcript levels which are involved in thyroid-mediated function. It also increased transcript levels related to oxidative stress, including hsp70, ahr1a, cyp1a, gpx and cat. Lastly, CYN exposure increased aqp3a and decreased dab2, which are involved in osmoregulation with a threshold of 10 µg/L. The present study demonstrates multiple effects of exposure to environmentally relevant CYN concentrations in zebrafish embryos.

Pre-activated antiviral innate immunity in the upper airways controls early SARS-CoV-2 infection in children.

Children have reduced severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection rates and a substantially lower risk for developing severe coronavirus disease 2019 compared with adults. However, the molecular mechanisms underlying protection in younger age groups remain unknown. Here we characterize the single-cell transcriptional landscape in the upper airways of SARS-CoV-2-negative (n = 18) and age-matched SARS-CoV-2-positive (n = 24) children and corresponding samples from adults (n = 44), covering an age range of 4 weeks to 77 years. Children displayed higher basal expression of relevant pattern recognition receptors such as MDA5 (IFIH1) and RIG-I (DDX58) in upper airway epithelial cells, macrophages and dendritic cells, resulting in stronger innate antiviral responses upon SARS-CoV-2 infection than in adults. We further detected distinct immune cell subpopulations including KLRC1 (NKG2A)+ cytotoxic T cells and a CD8+ T cell population with a memory phenotype occurring predominantly in children. Our study provides evidence that the airway immune cells of children are primed for virus sensing, resulting in a stronger early innate antiviral response to SARS-CoV-2 infection than in adults.

Cylindrospermopsin directly disrupts spermatogenesis in isolated male zebrafish testis.

Cylindrospermopsin (CYN) is a cytotoxin, and its documented effects in mammals include damage to several organs. CYN also has hormone-disrupting properties, including estrogenic activity, progesterone production inhibition, and apoptosis induction. While CYN has been reported to exert reproductive toxicity in mice, little is known about its effect on fish reproductive function. Using ex vivo organ culture, we investigated the direct action of CYN on the male reproductive system. Isolated zebrafish testis was exposed to 250, 500, and 1000 µg/L CYN for 24 h and 7 d, followed by histo-morphological analysis. The results demonstrate that exposure to CYN led to a decrease in cell types from all three phases of spermatogenesis in zebrafish testis. There were also significant changes in fshr, lhr, and igf3 transcript levels, as well as testosterone secretion following exposure to CYN. In summary, this study provides novel information on the adverse effects of CYN on testicular spermatogenesis and male reproduction in zebrafish. These results provide a framework for a better understanding of CYN toxicity and the mechanism underlying the adverse action of CYN on male reproduction in fish.

Fungal and bacterial coinfections increase mortality of severely ill COVID-19 patients.

BACKGROUND: SARS-CoV-2 predisposes patients to secondary infections; however, a better understanding of the impact of coinfections on the outcome of hospitalized COVID-19 patients is still necessary. AIM: To analyse death risk due to coinfections in COVID-19 patients. METHODS: The odds of death of 212 severely ill COVID-19 patients were evaluated, with detailed focus on the risks for each pathogen, site of infection, comorbidities and length of hospitalization. FINDINGS: The mortality rate was 50.47%. Fungal and/or bacterial isolation occurred in 89 patients, of whom 83.14% died. Coinfected patients stayed hospitalized longer and had an increased odds of dying (odds ratio (OR): 13.45; R2 = 0.31). The risk of death was increased by bacterial (OR: 11.28) and fungal (OR: 5.97) coinfections, with increased levels of creatinine, leucocytes, urea and C-reactive protein. Coinfections increased the risk of death if patients suffered from cardiovascular disease (OR: 11.53), diabetes (OR: 6.00) or obesity (OR: 5.60) in comparison with patients with these comorbidities but without pathogen isolation. The increased risk of death was detected for coagulase-negative Staphylococcus (OR: 25.39), Candida non-albicans (OR: 11.12), S. aureus (OR: 10.72), Acinetobacter spp. (OR: 6.88), Pseudomonas spp. (OR: 4.77), and C. albicans (OR: 3.97). The high-risk sites of infection were blood, tracheal aspirate, and urine. Patients with coinfection undergoing invasive mechanical ventilation were 3.8 times more likely to die than those without positive cultures. CONCLUSION: Severe COVID-19 patients with secondary coinfections required longer hospitalization and had higher risk of death. The early diagnosis of coinfections is essential to identify high-risk patients and to determine the right interventions to reduce mortality.

Cylindrospermopsin impairs tubular transport function in kidney cells LLC-PK1.

Cylindrospermopsin (CYN) has been involved in cases of poisoning in humans following ingestion. Studies have demonstrated that the kidney is the most affected organ. CYN exposure leads to low-molecular-weight proteinuria and increased excretions of the tubular enzymes in mice, suggesting the damage caused by CYN is mainly tubular. However, the mechanism involved in CYN nephrotoxicity remains unknown. Thus, in order to evaluate the effects of CYN exposure (0.1, 0.5 and 1.0 µg/mL) on tubular renal cells LLC-PK1 distinct mechanisms were analyzed by assessing cell death using flow cytometry, albumin uptake by fluorescence analysis, Na+/K+-ATPase activity by a colorimetric method, RT-qPCR of genes related to tubular transport and function as well as internalization of CYN by ELISA. In this study, CYN was found to induce necrosis in all concentrations. CYN also decreased albumin uptake as well as downregulated megalin and dab2 expression, both proteins involved in albumin endocytosis process. Moreover, CYN appears to be internalized by renal tubular cells through a receptor-mediated endocytosis. Finally, the present study demonstrates that CYN is responsible for disrupting tubular cell transport and function in LLC-PK1 cells.

High-Performance µ-Thermoelectric Device Based on Bi2Te3/Sb2Te3 p-n Junctions.

A flexible and ultralight planar thermoelectric generator based on 15 thermocouples composed of n-type bismuth telluride (Bi2Te3) and p-type antimony telluride (Sb2Te3) legs (each with 400 nm thick) connected in series, on 25 µm thick Kapton substrate, was fabricated with impressive power factor values of 2.7 and 0.8 mW K-2 m-1 (at 298 K) for Bi2Te3 and Sb2Te3 films, respectively. The p-n junction thermoelectric device can generate a maximum open-circuit voltage and output power of 210 mV and 0.7 µW (3.3 mW cm-2), respectively, for a temperature difference of 35 K, which is higher than the one observed for a conventional thermoelectric device with metallic contacts for p-n junctions. The results were combined with numerical simulations, showing a good match between the experimental and the numerical results. The current density versus voltage (J-V) characteristics of the fabricated p-n junctions revealed a diode behavior with a turn-on voltage of ≈0.3 V and an impressive rectifying ratio (I+1V/I-1V) of ≈2 × 104.

Fcγ-RI, Fcγ-RII and IL-10 as predictive biomarkers for post-therapeutic cicatrization time in monocytes from cutaneous leishmaniasis patients.

Cutaneous leishmaniasis (CL) treatment is based on therapy with Glucantime® , yet, there are few laboratory methods to monitor its success. In this study, ex vivo and in vitro evaluations of peripheral blood monocytes were performed in a longitudinal study to characterize the impact of Glucantime® on overall phenotypic/functional features of these cells from CL patients to identify predictive biomarkers for post-therapeutic monitoring by flow cytometry. The ex vivo evaluation from CL patients demonstrated a modulatory profile before treatment, with a decrease in TLR-2, FcγRII, HLA-DR, CD86, IFN-γR, TNF, IL-12, NO, and an increase in FcγRIII and IL-10R. Conversely, treatment changes some of these biomarker expressions by decreasing FcγRIII and IL-10R and increasing IFN-γR, IL-12 and NO. Moreover, an in vitro analysis of these patients showed a reduced phagocytic capacity of Leishmania braziliensis and higher levels of IL-10 and TGF-ß modulating functional profile. Regardless of the compromised L. braziliensis phagocytic capacity, treatment re-established the production of IL-12, IL-10, TGF-ß and NO at the basal level. Notably, monocytes from patients with early cicatrization showed enhanced FcγRI and FcγRII expressions and reduced IL-10, which was further corroborated by a baseline fold change analysis. Finally, the logistic regression model emphasized the performance of FcγRI, FcγRII and IL-10 as robust predictive biomarkers for post-therapeutic cicatrization during cutaneous leishmaniasis.

Renal tubular damage caused by cylindrospermopsin (cyanotoxin) in mice.

Cylindrospermopsin (CYN) is a cyanotoxin and a hydrophilic alkaloid of 415 Da. The principal effect of CYN is the inhibition of protein synthesis, and it can damage various organs. Studies have demonstrated that the kidney is the most affected organ. CYN has played roles in at least two poisoning cases, i.e., the mysterious Palm Island disease in Australia and the event at Caruaru in Brazil. Therefore, we aimed to determine how CYN disrupts the renal tissue. Dose-response curves following single intraperitoneal injections of purified CYN (at 0, 16, 32, 64 and 128 µg CYN/kg body weight) were created in 10-week-old male BALB/C mice (n = 4). Renal physiology parameters were analyzed after 7 and 14 days. However, no alterations in the glomerular filtration rate (GFR) or nephrin expression (a crucial protein for glomerular integrity) were observed. We detected low-molecular-weight proteinuria and increased excretions of the tubular enzymes lactate dehydrogenase (LDH) and gamma-glutamyl transferase (GGT) at doses of 16, 32 and 64 µg CYN/kg body weight. Furthermore, we observed increases in the renal interstitial space and collagen deposition that indicated edema and fibrosis. The data seem to indicate that the damage is in the proximal tubule.

Cylindrospermopsin effects on cell viability and redox milieu of Neotropical fish Hoplias malabaricus hepatocytes.

Cylindrospermopsin (CYN) is a cyanotoxin that is cytotoxic to a wide variety of cells, particularly to the hepatocytes. In this study, the toxic effects of purified CYN were investigated in primary cultured hepatocytes of Neotropical fish Hoplias malabaricus. After isolation, attachment, and recovery for 72 h, the cells were exposed for 72 h to 0, 0.1, 1.0, 10, and 100 µg l-1 of CYN. Then, cell viability and a set of oxidative stress biomarker responses were determined. Catalase, superoxide dismutase, glucose-6-phosphate dehydrogenase, and glutathione S-transferase activities were not affected by exposure to CYN. Concentration-dependent decrease of glutathione reductase activity occurred for most CYN-exposed groups, whereas non-protein thiol content increased only for the highest CYN concentration. Lipid peroxidation, protein carbonylation, and DNA damage levels were not altered, but reactive oxygen species levels increased in the cells exposed to the highest concentration of CYN. Cell viability decreased in all the groups exposed to CYN. Thus, CYN may cause a slight change in redox balance, but it is not the main cause of cell death in H. malabaricus hepatocytes.

Short communication: Latin-style fresh cheese enhances lactic acid bacteria survival but not Listeria monocytogenes resistance under in vitro simulated gastrointestinal conditions.

Different studies in humans have provided evidence about the health benefits of probiotics. However, most probiotic strains do not maintain good viability in the harsh conditions of the gastrointestinal tract (GIT). In the present study, Latin-style fresh cheese produced with potential probiotic bacteria was tested to evaluate this cheese type as a food carrier for the delivery of viable microorganisms after exposure to simulated GIT conditions. The resistance of 28 lactic acid bacteria (LAB) strains and Listeria monocytogenes upon exposure to acidic conditions (pH 2.5) and bile and pancreatic enzymes (0.3% of bile salts and 0.1% of pancreatin) was evaluated in vitro. When compared with fresh cultures, fresh cheese greatly improved LAB survival to simulated GIT conditions, as no loss of viability was observed in either acidic conditions (pH 2.5) or bile salts and pancreatin environment over a 3-h period. In opposition, L. monocytogenes did not survive after 1h under acidic conditions. These data demonstrated that Latin-style fresh cheese could play an important role in probiotic protection against gastrointestinal juices, enhancing delivery within the gut and thereby maximizing potential health benefits of LAB.

Protective Profile Involving CD23/IgE-mediated NO Release is a Hallmark of Cutaneous Leishmaniasis Patients from the Xakriabá Indigenous Community in Minas Gerais, Brazil.

In this study, we described, for the first time, specific aspects of an anti-Leishmania immune response in a Brazilian Xakriabá indigenous community. Induction of an intracellular NO pathway, triggered by the binding of IgE to CD23 receptor in IFN-γ/IL-4 cytokines environment, was evaluated in localized cutaneous leishmaniasis (LCL) carriers and positive Montenegro skin test (MST) individuals without skin lesion (MT(+) SL(-)). Our data demonstrated that the higher frequency of CD23(+) CD14(+) monocytes and the increased serum levels of IgE observed in the LCL group were even higher in LCL carriers with late lesions (LCL≥60). Furthermore, patients with LCL presented increased NO production after Leishmania (Viannia) braziliensis stimulation and this NO profile was independent of the time of the lesion (recent LCL<60 or late LCL≥60). We also showed that the increased frequency of IFN-γ(+) and IL-4(+) CD4(+) T cells is related to the MT(+) SL(-) group. The results of biomarker signature curves demonstrated that in the MT(+) SL(-) group, the index signature was characterized by DAF-2T(+) CD14(+)/IL-4(+) CD8(+)/IFN-γ(+) CD4(+)/IL-4(+) CD4(+). On the other hand, the LCL group presented a higher index of DAF-2T(+) CD14(+)/CD23(+) CD14(+)/IL-4(+) CD8(+), associated with a lower index of IFN-γ(+) CD8(+). Considering the time of lesion, data analysis demonstrated that the main differences observed were highlighted in LCL<60 patients, with a higher index of CD23(+) CD14(+), which was also present in LCL≥60 patients. In conclusion, our data suggest that the protective immune response involving CD23-IgE-mediated NO release is a hallmark of patients with LCL. However, in MT(+) SL(-) individuals, another different leishmanicidal mechanism seems to be involved.

Etiological treatment of Chagas disease patients with benznidazole lead to a sustained pro-inflammatory profile counterbalanced by modulatory events.

In the present study, we characterized the phagocytic capacity, cytokine profile along with the FCγ-R and TLR expression in leukocytes from Chagas disease patients (indeterminate-IND and cardiac-CARD) before and one-year after Bz-treatment (INDT and CARDT). A down-regulation of IL-17, IFN-γ and IL-10 synthesis by neutrophils was observed in CARDT. The Bz-treatment did not impact on the expression of phagocytosis-related surface molecules or monocyte-derived cytokine profile in INDT. Although CARDT showed unaltered monocyte-phagocytic capacity, up-regulated expression of Fcγ-RI/III and TLR-4 may be related to their ability to produce IL-10 and TGF-ß. Down-regulation of lymphocyte-derived cytokine was observed in INDT whereas up-regulated cytokine profile was observed for lymphocytes in CARDT. Analysis of cytokine network revealed that IND displayed a multifaceted cytokine response characterized by strong connecting axes involving pro-inflammatory/regulatory phagocytes and lymphocytes. On the other hand, CARD presented a modest cytokine network. The Bz-treatment leads to distinct cytokine network: decreasing the links in INDT, with a pivotal role of IL-10(+) monocytes and expanding the connections in CARDT. Our findings highlighted that the Bz-treatment contributes to an overall immunomodulation in INDT and induces a broad change of immunological response in CARDT, eliciting an intricate phenotypic/functional network compatible with beneficial and protective immunological events.

In vitro tests of resveratrol radiomodifying effect on rhabdomyosarcoma cells by comet assay.

Cancer is a global public health problem. Resveratrol is a defensive polyphenol that is synthesized by a wide variety of plants in response to exposure to ultraviolet radiation or also due to mechanical stress caused by the action of pathogens and chemical and physical agents. Grape vines have a high capacity to produce resveratrol, so grape juice and wine, mainly red wine, are considered good sources of resveratrol. The protective effects of resveratrol include promotion of antiinflammatory response, antitumor activity and prevention of degenerative diseases, reduced incidence of cardiovascular diseases and inhibition of platelet aggregation, among others. Therefore, resveratrol is considered to be a cell protector. However, at high concentrations, resveratrol promotes contrary effects by sensitizing cells. The aim of this study was to investigate in vitro the radiomodifying effect of resveratrol in culture of human rhabdomyosarcoma cells (RD) by applying the comet assay to evaluate the cell damage and repair capacity. The LD50 (lethal dose) obtained was 499.95 ± 9.83 Gy (Mean ± SD) and the CI50 (cytotoxicity index) was 150 µM in the RD cells. Based on these data, it was defined the gamma radiation doses (50 and 100 Gy) and resveratrol concentrations (15, 30 and 60 µM) to be used in this study. The results indicated that resveratrol acts as a cell protector at a concentration of 15 µM and has a cytotoxic effect at 60 µM. However, with the interaction of the gamma radiation, the concentration of 60 µM did not produce a statistically significant radiosensitizing effect.

Evaluation of multiplex PCR in first episodes of febrile neutropenia as a tool to improve early yeast diagnosis in leukemic/preleukemic patients.

In febrile neutropenic onco-hematological patients, delayed microbiological diagnosis leads to an increase in morbidity and mortality. Identification of the microorganism changes antibiotic therapy in more than half of cases; however, in only 20-30 % of such cases pathogen isolation is achieved. This study evaluates the frequency of fungus infection and its etiology in onco-hematological patients with febrile neutropenia utilizing blood cultures and non-commercial multiplex polymerase chain reaction (MT-PCR) primers. Fifty-three febrile neutropenia episodes in 35 onco-hematological patients were observed, and the results for the first unique 30 episodes are presented. Blood cultures were positive for Candida tropicalis (one case), gram-positive bacteria (two cases), and gram-negative bacteria (four cases), showing a 23.3 % microbiological isolation rate. Multiplex-PCR pan-fungal sequence was positive in 18 cases (60 %), and further sequencing identified fugal pathogens in 11 cases (Candida glabrata and Candida parapsilosis being the most common). MT-PCR pan-fungal sequence amplification was detected in 13 of 16 patients that later received antifungal treatment for clinical reasons only, while positivity was found in 5 out of 14 patients that did not receive antifungal treatment (p = 0.02). These results show that performing in-house non-commercial MT-PCR is feasible and may provide additional information about fungal infection without the need to wait for culture results. Further research is necessary to incorporate this technology into the decision-making process.

High prevalence of asymptomatic Leishmania spp. infection among liver transplant recipients and donors from an endemic area of Brazil.

Visceral leishmaniasis is an uncommon disease in transplant recipients; however, if left untreated, the mortality can be high. If an organ donor or recipient is known to be an asymptomatic Leishmania spp. carrier,monitoring is advised. This study proposes to assess the prevalence of asymptomatic Leishmania spp.infection in liver transplant donors and recipients from an endemic area. A total of 50 liver recipients and 17 liver donors were evaluated by direct parasite search, indirect fluorescent antibody test (IFAT), anti-Leishmania rK39 rapid test and Leishmania spp.DNA detection by polymerase chain reaction (PCR).Leishmania spp. amastigotes were not observed in liver or spleen tissues. Of the 67 serum samples, IFAT was reactive in 1.5% and indeterminate for 17.9%, and the anti-Leishmania rK39 rapid test was negative for all samples. The PCR test was positive for 7.5%, 8.9%, and 5.9% of blood, liver and spleen samples, respectively(accounting for 23.5% of the donors and 8% of the recipients). Leishmania infantum-specific PCR confirmed all positive samples. In conclusion, a high prevalence of asymptomatic L. infantum was observed in donors and recipients from an endemic area, and PCR was the most sensitive method for screening these individuals.

Método de amostragem e caracterização química da forragem consumida por bovinos em pasto consorciado de aveia e azevém / Method of sampling and chemical characterization of forage intake by cattle in pasture ryegrass intercropping oats

O presente ensaio foi conduzido com o objetivo de se estudarem diferentes métodos de amostragem de pasto e de se estimar a composição química da dieta consumida por novilhos Holandeses, mantidos em pastagem consorciada de aveia preta (Avena strigosa Schreb) e azevém (Lolium multiflorum Lam). Os métodos avaliados foram o corte da forragem rente ao solo (CFRS), o pastejo simulado (PSI) e a coleta de extrusa ruminal (CERU). Os teores médios de proteína bruta (PB), nutrientes digestíveis totais (NDT), fibra em detergente neutro (FDN) e fibra em detergente ácido (FDA) foram de 9,7; 62,2; 64,5 e 33,1% para o tratamento CFRS; de 9,8; 65,4; 59,6 e 30,0% para o PSI, e de 11,4; 70,8; 51,6 e 25,5% para CERU, respectivamente. Não houve diferença estatística entre os métodos CFRS e PSI, todavia, em relação ao método CERU, ambos subestimaram a concentração proteica e a energética, e superestimaram a quantidade de parede celular presente na dieta. Conclui-se que a coleta da extrusa ruminal pode ser um método adequado para caracterizar a dieta consumida por novilhos em pastagem consorciada de aveia e azevém.

The experiment was conducted to study different methods of pasture sampling, to estimate the chemical composition of the diet of Holstein steer, and grazing pasture of oat (Avena strigosa Schreb)and ryegrass (Lolium multiflorum Lam). The methods evaluated were Clipping Close by Soil (CCS), Hand-Plucking (HPL) and Rumen Evacuation (REV). The averages for crude protein (CP), total digestible nutrients (TDN), neutral detergent fiber (NDF) and acid detergent fiber (ADF) were 9.7, 62.2, 64.5 and 33.1% for the CCS treatment; 9.8, 65.4, 59.6 and 30.0% for HPL, and 11.4, 70.8, 51.6 and 25.5% for REV, respectively. There was no statistical difference between CCS and HPL methods, however, in relation to the REV method, both underestimated protein and energy concentration, and overestimated the amount of cell walls in the diet. The conclusion is that rumen evacuation may be an adequate method to characterize the diet consumed by steers on oats and ryegrass pastures.

Distinct pattern of immunophenotypic features of innate and adaptive immunity as a putative signature of clinical and laboratorial status of patients with localized cutaneous leishmaniasis.

In this study, we have analysed the phenotypic features of innate/adaptive immunity of patients with localized cutaneous leishmaniasis (LCL), categorized according to their clinical/laboratorial status, including number of lesion (L1; L2­4), days of illness duration (≤60;>60) and positivity in the Montenegro skin test (MT−;MT+). Our findings highlighted a range of phenotypic features observed in patients with LCL (↑%HLA-DR+ neutrophils; ↑CD8+ HLA-DR+/CD4+ HLA-DR+ T cell ratio; ↑HLA-DR in B lymphocytes, ↑%CD23+ neutrophils, monocytes and B cells; ↑α-Leishmania IgG and ↑serum NO2⁻ + NO3⁻). Selective changes were observed in L1 (↑%HLA-DR+ neutrophils, ↑CD8+ HLA-DR+/CD4+ HLA-DR+ T cell ratio and ↑serum NO2⁻ + NO3⁻) as compared to L2­4 (↑%CD5− B cells; ↑CD23+ B cells and ↑α-Leishmania IgG). Whilst ≤60 presented a mixed profile of innate/adaptive immunity (↓%CD28+ neutrophils and ↑%CD4+ T cells), >60 showed a well-known leishmanicidal events (↑CD8+ T cells; ↑serum NO2⁻ + NO3⁻ and ↑α-Leishmania IgG). MT+ patients showed increased putative leishmanicidal capacity (↑%HLA-DR+ neutrophils; ↑%CD23+ monocytes; ↑CD8+ HLA-DR+/CD4+ HLA-DR+ T cell ratio and ↑ serum NO2⁻ + NO3⁻). Overall, a range of immunological biomarkers illustrates the complex immunological network associated with distinct clinical/laboratorial features of LCL with applicability in clinical studies.

Impaired phagocytic capacity driven by downregulation of major phagocytosis-related cell surface molecules elicits an overall modulatory cytokine profile in neutrophils and monocytes from the indeterminate clinical form of Chagas disease.

The distinct ability of phagocytes to present antigens, produce cytokines and provide co-stimulatory signals may contribute to the severity of the outcome of Chagas disease. In this paper, we evaluate the phenotypic features of phagocytes along with the cytokine signature of circulating T-cells from Chagas disease patients with indeterminate (IND) and cardiac (CARD) clinical forms of the disease. Our data demonstrated that neutrophils from IND patients displayed an impaired ability to produce cytokines. A lower Trypanosoma cruzi phagocytic index and higher nitric oxide levels were characteristics of monocytes from IND. The impaired phagocytic capacity did not reflect on the levels of anti-T. cruzi IgG, but was detectable in the downregulation of Fc-γR, TLR and CR1 molecules. The monocyte-derived cytokine signature demonstrated that a down-regulated synthesis of IL-12 and a modulatory state were evidenced by a positive correlation between IL-12 and IL-10 with a lower synthesis of TNF-α. The down-regulation of MHC-II and CD86 in monocytes supports the occurrence of particularities in the APC-activation-arm in IND, and may be involved in the T-cell pro-inflammatory pattern counterbalanced by a potent IL-10 response. Our findings support the hypothesis that differential phenotypic features of monocytes from IND may be committed to the induction of a distinct immune response related to low morbidity in chronic Chagas disease.