Fernanda de Freitas Anibal

Leishmaniasis and Chagas disease chemotherapy: a critical review

Leishmaniasis and Chagas disease, caused by the kinetoplastid parasites Leishmania spp and Trypanosoma cruzi, respectively, are among the most important parasitic diseases, affecting millions of people and considered to be within the most relevant group of neglected tropical diseases. Chemotherapy is the main alternative to control such parasites, nevertheless, current treatments are far from satisfactory. This review outlines the current understanding on different drugs against leishmaniasis and Chagas disease and their mechanism of action. Recent approaches in the area of anti-leishmanial and trypanocidal therapies are also enumerated, as well as the search for new drugs.

Anti-Inflammatory Properties of Menthol and Menthone in Schistosoma mansoni Infection

Schistosomiasis is a parasitic disease caused by several species of trematode worms and it is believed that more than 261 million people are affected worldwide. New drug development has become essential because there is a risk of the parasite becoming resistant to Praziquantel, the only drug available for this infection. This study evaluated parasitological, immunological and histological parameters in mice infected with Schistosoma mansoni and treated with an herbal commercial medicine. This drug consists of menthol (30–55%) and menthone (14–32%). A 60 day treatment regimen with the herbal medicine decreased the number of S. mansoni eggs in the feces, liver, and intestine and reduced the number of hepatic granulomas. We observed a reduction of 84% in blood eosinophilia and a decrease in the IL-4 and IL-10 blood levels after treatment. Therefore, we propose that schistosomiasis treatment with this herbal medicine for 60 days has an immunomodulatory and anti-inflammatory action in this animal model for schistosomiasis thus contributing to the decrease in physio pathological effects caused by S. mansoni infection.

Morphological Characteristics of Schistosoma mansoni PZQ-Resistant and -Susceptible Strains Are Different in Presence of Praziquantel

Schistosomiasis is one of the most common human parasitic diseases whose socioeconomic impact is only surpassed by malaria. Praziquantel (PZQ) is the only drug commercially available for the treatment of all schistosome species causing disease in humans. However, there has been stronger evidences of PZQ-resistance on Schistosoma mansoni and thus it is very important to study the phenotypic characteristics associated with it. The aim of this study was to evaluate morphological alterations in S. mansoni PZQ-resistant adult worms and eggs, by comparing a PZQ- resistant strain obtained under PZQ drug pressure with a PZQ-susceptible strain. For this, scanning electronic microscopy was used to assess tegumental responsiveness of both strains under PZQ exposure, and optical microscopy allowed the monitoring of worms and eggs in the presence of the drug. Those assays showed that PZQ-susceptible worms exposed to the drug had more severe tegumental damages than the resistant one, which had only minor alterations. Moreover, contrary to what occurred in the susceptible strain, resistant worms were viable after PZQ exposure and gradually regaining full motility after removal of the drug. Eggs from resistant strain parasites are considerably smaller than those from susceptible strain. Our results suggest that there might be a difference in the tegument composition of the resistant strain and that worms are less responsive to PZQ. Changes observed in egg morphology might imply alterations in the biology of schistosomes associated to PZQ-resistance, which could impact on transmission and pathology of the disease. Moreover, we propose a hypothetical scenario where there is a different egg tropism of the S. mansoni resistant strain. This study is the first comparing two strains that only differ in their resistance characteristics, which makes it a relevant step in the search for resistance determinants.

Immunological and parasitological parameters in Schistosoma mansoni-infected mice treated with crude extract from the leaves of Mentha x piperita L.

Schistosomiasis is a chronic disease caused by an intravascular trematode of the genus Schistosoma. Praziquantel is the drug used for treatment of schistosomiasis; nevertheless failure of treatment has been reported. Consequently, the identification of new effective schistosomicidal compounds is essential to ensure the effective control of schistosomiasis in the future. In this work we investigated the immunomodulatory and antiparasitic effects of the crude leaves extract of Mentha x piperita L. (peppermint) on murine Schistosomiasis mansoni. Female Balb/c mice were infected each with 50 S. mansoni cercariae and divided into three experimental groups: (I) untreated; (II) treated daily with M. x piperita L. (100mg/kg) and III) treated on 1/42/43 days post-infection with Praziquantel (500mg/kg). Another group with uninfected and untreated mice was used as a control. Subsequently, seven weeks post-infection, S. mansoni eggs were counted in the feces, liver and intestine. Worms were recovered by perfusion of the hepatic portal system and counted. Sera levels of IL-10, IL-5, IL-13, IFN-γ, IgG1, IgE and IgG2a were assayed by ELISA. Animals treated with a daily dose of M. x piperita L. showed increased sera levels of IL-10, IFN-γ, IgG2a and IgE. Besides, M. x piperita L. treatment promoted reduction in parasite burden by 35.2% and significant decrease in egg counts in the feces and intestine.

Experimental type 2 diabetes induction reduces serum vaspin, but not serum omentin, in Wistar rats

Vaspin and omentin are adipose tissue adipokines that have often been related to obesity and its comorbidities. The aim of this study was to investigate the behaviour of serum omentin and vaspin in models of type 2 diabetes. To do this, Wistar rats (~200 g) were randomly divided into two groups: a non‐diabetic group (n = 6) and a diabetic group fed on a high‐fat diet (n = 6) and a low dose of streptozotocin (Sigma®). All procedures were approved by the Brazilian Ethics Committee. Body weight (BW) and food intake were recorded daily. Tail blood glucose levels were assessed at the end of the diabetes induction period. The insulin tolerance test (ITT) was performed after the diabetes induction period (7 weeks). The serum and tissues (liver, pancreas, and retroperitoneal (RET), epididymal (EPI) and visceral (VIS) white adipose tissues) were immediately removed and weighed. Analyses of levels of insulin, omentin, vaspin, adiponectin and inflammatory cytokines IL‐6, IL‐8 (CXCL8), TNF‐α and C‐reactive protein (CRP) in serum were performed using the enzyme‐linked immunosorbent assay (ELISA). Our results showed that IL‐8 and CRP serum levels in the diabetic group were significantly higher than in the non‐diabetic group. Vaspin and adiponectin values were lower for the diabetic group than for the non‐diabetic group. Omentin, IL‐6 and TNF‐α values did not differ between the groups. Our results showed that both the metabolism of the adipose tissue and the secretion of adipokines may be affected in diabetic rats. Omentin showed no difference between the groups, although the vaspin values decreased in the diabetic group.

Laser therapy modulates systemic inflammatory processes and muscle atrophy in an experimental model of sepsis in rats

Abstract Objective: The aim of this study was to determine the effectiveness of low-level laser therapy (LLLT) on the modulation of the systemic inflammatory processes and skeletal muscle morphology in an experimental sepsis model (cecal ligation and puncture, CLP). Study design: Seventy-two male Wistar rats were randomly divided into three groups: control group (CG); sepsis group (SG) where rats were submitted to CLP but without LLLT treatment, and the sepsis laser-treated group (SLG). Laser irradiation (GaAlAs laser, continuous wave, 808 nm, 30 mW, 48 s, 30 J/cm2, 0.028 cm2, 1.07 mW/cm2) was performed immediately after surgery and every 24 h at 4 points (on the middle of tibialis anterior and diaphragm, bilaterally), through the punctual contact technique. All sepsis animals were sacrificed at 6, 24, 48 and 72 h post-surgery. The immunohistochemistry analysis was used to verify the expression of proteins related to the regulation of muscle wasting (MuRF-1 and atrogin). In order to investigate the action of LLLT on inflammatory mediators in the rat sepsis model, two inflammatory cytokines IL-6 and IL-10, were evaluated. Results: The results showed that the laser-treated animals presented a lower IL-6 activity and decreased atrogin and MuRF-1 immunoexpression. However, no difference was observed in muscle cross-sectional area between the experimental groups. Conclusion: These results suggest that LLLT was able to decrease the systemic inflammation and muscle atrophy markers, preventing muscle protein degradation.

Toxocara canis and the allergic process

The protective effect of infectious agents against allergic reactions has been thoroughly investigated. Current studies have demonstrated the ability of some helminths to modulate the immune response of infected hosts. The objective of the present study was to investigate the relationship between Toxocara canis infection and the development of an allergic response in mice immunised with ovalbumin (OVA). We determined the total and differential blood and bronchoalveolar lavage fluid cells using BALB/c mice as a model. To this end, the levels of interleukin (IL)-4, IL-5 and IL-10 and anti-OVA-IgE were measured using an ELISA. The inflammatory process in the lungs was observed using histology slides stained with haematoxylin and eosin. The results showed an increase in the total number of leukocytes and eosinophils in the blood of infected and immunised animals at 18 days after infection. We observed a slight lymphocytic inflammatory infiltrate in the portal space in all infected mice. Anti-OVA-IgE levels were detected in smaller proportions in the plasma of immunised and infected mice compared with mice that were only infected. Therefore, we concluded that T. canis potentiates inflammation in the lungs in response to OVA, although anti-OVA-IgE levels suggest a potential reduction of the inflammatory process through this mechanism.

Comparative Proteomics Reveals Characteristic Proteins on Praziquantel-resistance in Schistosoma mansoni

The extensive use of Praziquantel (PZQ), the only drug available to treat schistosomiasis, has brought concern about the emergence of PZQ-resistance/tolerance by Schistosoma spp., thus reaffirming an urge for the development of new treatment alternatives. Therefore, it is imperative and urgent to study this phenomenon trying to understand what is involved in its occurrence. Studies of Schistosoma spp. genome, transcriptome and proteome are crucial to better understand this situation. By stepwise drug pressure from a fully susceptible parasite strain, our group selected a S. mansoni variant strain stably resistant to PZQ and isogenic to its fully susceptible parental counterpart, except for the genetic determinants of PZQ-resistance phenotype. Based on this, the objective of this study was to compare the proteomes of both strains, identifying proteins from male and female adult worms of PZQ-resistant and PZQ-susceptible strains, exposed and not exposed to PZQ, which were separated by high-resolution two-dimensional electrophoresis and sequenced by high throughput LC-MS/MS. Likewise, this work is extremely relevant since for the first time the proteome of a S. mansoni PZQ-resistant strain is studied and compared to the proteome of the respective S. mansoni PZQ-susceptible strain. This study identified 60 S. mansoni proteins, some of which differentially expressed in either strain, which may putatively be involved in the PZQ-resistance phenomenon. This information represents substantial progress towards deciphering the worm proteome. Furthermore, these data may constitute an informative source for further investigations into PZQ-resistance and increase the possibility of identifying proteins related to this condition, possibly contributing to avoid or decrease the likelihood of development and spread of PZQ-resistance. This is an innovative study that opens doors to PZQ-resistance surveys, contributing to discover a solution to PZQ-resistance problem, as suggests new potential targets for study.

Photodynamic inactivation using curcuminoids and Photogem on caenorhabditis elegans

Resistance to various anthelmintic drugs is reported in many animals and can become a severe problem for human and animal health. In this study, Photogem® and three curcuminoids compounds (curcumin, demethoxycurcumin, bisdemethoxycurcumin) were used as photosensitizers in the photodynamic inactivation (PDI) in the helminth model Caenorhabditis elegans to investigate the ability of this procedure to worm life cycle. Initially, the presence and location of the photosensitizers in the worms body were verified by fluorescence confocal microscopy. Curcumin was deposited in the digestive tract and Photogem® along the body of the animal in the incubation time of 12 hours with the photosensitizer. Subsequently, a PDI procedure using a LED device was performed to illuminate the worms treated with the photosensitizers. The worms were observed by optical microscopy until 48 hours after the PDI to verify the changes in motility, the presence of eggs and larvae and the number of live worms. Curcuminoids tested separately and in combination and two light doses of 30 J/m2 no changes were observed in the life cycle of the worm at concentrations of 2 mM and 1 mM. However, in treatment with Photogem® and a light dose of 100 J/m2 a reduction in motility and reproduction of the worm with 0.2 mg/mL was observed after 6 hours of exposure, in addition to the death of most worms at concentrations of 6, 4, and 2 mg/mL. We suggest, therefore, that photodynamic inactivation with Photogem® may present an anthelmintic effect against C. elegans, but there is a need for studies on helminths with parasitic activity.

Photobiomodulation on critical bone defects of rat calvaria: a systematic review

Bone defects following trauma represent a high impact on the quality of life of millions of people around the world. The aim of this study was to review photobiomodulation (PBM) action in the treatment of bone critical defects in rat calvaria, related to evaluation of the current protocols applied. One hundred and forty-seven articles related to the subject were found by searching the main databases (Pubmed, Lilacs, Web of Science, and Scopus) considering the period of publication until the year 2017, and only 14 corresponded the inclusion criteria established for this systematic review. The main parameters of the PBM were expressed in Table 1. In addition, it was possible to observe the use of two different wavelengths (red and infrared), which are considered therapeutic. Most of the evaluated articles presented positive results that describe a greater amount of neoformed bone, an increase in collagen synthesis, and a contribution to microvascular reestablishment. However, two studies report no effect on the repair process when the PBM was used. In addition, we observed considerable variations between the values of power, fluence, and total energy, which make it difficult to compare the results presented between the selected studies. It was possible to conclude that the infrared laser was more effective in positively stimulating the bone repair process of critical defects. Furthermore, a discrepancy was found in the parameter values used, which made it difficult to choose the best protocol for the treatment of this type of lesion.