Lisdexamfetamine dimesylate-exposition in male rats during the peripubertal period impairs inflammatory mechanisms, antioxidant activity, and apoptosis process in kidneys of male pubertal rats.

Lisdexamfetamine dimesylate (LDX) is a prodrug of dextroamphetamine, which has been widely recommended for the treatment of Attention-Deficit/Hyperactivity Disorder (ADHD). There are still no data in the literature relating the possible toxic effects of LDX in the kidney. Therefore, the present study aims to evaluate the effects of LDX exposure on morphological, oxidative stress, cell death and inflammation parameters in the kidneys of male pubertal Wistar rats, since the kidneys are organs related to the excretion of most drugs. For this, twenty male Wistar rats were distributed randomly into two experimental groups: LDX group-received 11,3 mg/kg/day of LDX; and Control group-received tap water. Animals were treated by gavage from postnatal day (PND) 25 to 65. At PND 66, plasma was collected to the biochemical dosage, and the kidneys were collected for determinations of the inflammatory profile, oxidative status, cell death, and for histochemical, and morphometric analyses. Our results show that there was an increase in the number of cells marked for cell death, and a reduction of proximal and distal convoluted tubules mean diameter in the group that received LDX. In addition, our results also showed an increase in MPO and NAG activity, indicating an inflammatory response. The oxidative status showed that the antioxidant system is working undisrupted and avoiding oxidative stress. Therefore, LDX-exposition in male rats during the peripubertal period causes renal changes in pubertal age involving inflammatory mechanisms, antioxidant activity and apoptosis process.

Doxorubicin causes cachexia, sarcopenia, and frailty characteristics in mice.

While chemotherapy treatment can be lifesaving, it also has adverse effects that negatively impact the quality of life. To investigate the effects of doxorubicin chemotherapy on body weight loss, strength and muscle mass loss, and physical function impairments, all key markers of cachexia, sarcopenia, and frailty. Seventeen C57/BL/6 mice were allocated into groups. 1) Control (n = 7): mice were exposed to intraperitoneal (i.p.) injections of saline solution. 2) Dox (n = 10): mice were exposed to doxorubicin chemotherapy cycles (total dose of 18 mg/kg divided over 15 days). The body weight loss and decreased food intake were monitored to assess cachexia. To assess sarcopenia, we measured muscle strength loss using a traction method and evaluated muscle atrophy through histology of the gastrocnemius muscle. To evaluate physical function impairments and assess frailty, we employed the open field test to measure exploratory capacity. Doxorubicin administration led to the development of cachexia, as evidenced by a significant body weight loss (13%) and a substantial decrease in food intake (34%) over a 15-day period. Furthermore, 90% of the mice treated with doxorubicin exhibited sarcopenia, characterized by a 20% reduction in traction strength (p<0,05), a 10% decrease in muscle mass, and a 33% reduction in locomotor activity. Importantly, all mice subjected to doxorubicin treatment were considered frail based on the evaluation of their overall condition and functional impairments. The proposed model holds significant characteristics of human chemotherapy treatment and can be useful to understand the intricate relationship between chemotherapy, cachexia, sarcopenia, and frailty.

Trilobolide-6-O-isobutyrate from Sphagneticola trilobata acts by inducing oxidative stress, metabolic changes and apoptosis-like processes by caspase 3/7 activation of human lung cancer cell lines.

BACKGROUND: Lung cancer, a chronic and heterogeneous disease, is the leading cause of cancer-related death on a global scale. Presently, despite a variety of available treatments, their effectiveness is limited, often resulting in considerable toxicity and adverse effects. Additionally, the development of chemoresistance in cancer cells poses a challenge. Trilobolide-6-O-isobutyrate (TBB), a natural sesquiterpene lactone extracted from Sphagneticola trilobata, has exhibited antitumor effects. Its pharmacological properties in NSCLC lung cancer, however, have not been explored. PURPOSE: This study evaluated the impact of TBB on the A549 and NCI-H460 tumor cell lines in vitro, examining its antiproliferative properties and initial mechanisms of cell death. METHODS: TBB, obtained at 98 % purity from S. trilobata leaves, was characterized using chromatographic techniques. Subsequently, its impact on inhibiting tumor cell proliferation in vitro, TBB-induced cytotoxicity in LLC-MK2, THP-1, AMJ2-C11 cells, as well as its effects on sheep erythrocytes, and the underlying mechanisms of cell death, were assessed. RESULTS: In silico predictions have shown promising drug-likeness potential for TBB, indicating high oral bioavailability and intestinal absorption. Treatment of A549 and NCI-H460 human tumor cells with TBB demonstrated a direct impact, inducing significant morphological and structural alterations. TBB also reduced migratory capacity without causing toxicity at lower concentrations to LLC-MK2, THP-1 and AMJ2-C11 cell lines. This antiproliferative effect correlated with elevated oxidative stress, characterized by increased levels of ROS, superoxide anion radicals and NO, accompanied by a decrease in antioxidant markers: SOD and GSH. TBB-stress-induced led to changes in cell metabolism, fostering the accumulation of lipid droplets and autophagic vacuoles. Stress also resulted in compromised mitochondrial integrity, a crucial aspect of cellular function. Additionally, TBB prompted apoptosis-like cell death through activation of caspase 3/7 stressors. CONCLUSION: These findings underscore the potential of TBB as a promising candidate for future studies and suggest its viability as an additional component in the development of novel anticancer drugs prototypes.

Biogenic silver nanoparticle exhibits schistosomicidal activity in vitro and reduces the parasitic burden in experimental schistosomiasis mansoni.

Schistosomiasis is a neglected tropical parasitic disease that affects millions of people, being the second most prevalent parasitic disease worldwide. The current treatment has limited effectiveness, drug-resistant strains, and is not effective in different stages of the disease. This study investigated the antischistosomal activity of biogenic silver nanoparticles (Bio-AgNp) against Schistosoma mansoni. Bio-AgNp presented direct schistosomicidal activity on newly transformed schistosomula causing plasma membrane permeabilization. In S. mansoni adult worms, reduced the viability and affected the motility, increasing oxidative stress parameters, and inducing plasma membrane permeabilization, loss of mitochondrial membrane potential, lipid bodies accumulation, and autophagic vacuoles formation. During the experimental schistosomiasis mansoni model, Bio AgNp restored body weight, reduced hepatosplenomegaly, and decrease the number of eggs and worms in feces and liver tissue. The treatment also ameliorates liver damage and reduces macrophage and neutrophil infiltrates. A reduction in count and size was evaluated in the granulomas, as well as a change to an exudative-proliferative phase, with a local increase of IFN-γ. Together our results showed that Bio-AgNp is a promising therapeutic candidate for studies of new therapeutic strategies against schistosomiasis.

Solidagenone in vivo leishmanicidal activity acting in tissue repair response, and immunomodulatory capacity in Leishmania amazonensis.

Leishmaniasis is a group of chronic parasitic diseases in humans caused by species of the Leishmania genus. Current treatments have high toxicity, cost, duration, limited effectiveness, significantly complex administration, and drug-resistant strains. These factors highlight the importance of research into new therapies that use drugs without toxic effects. Solidagenone (SOL), the main labdane diterpene isolated from the plant Solidago chilensis, has anti-inflammatory, gastroprotective, antioxidant, tissue repair-inducing effects, suggesting a role in novel drug development. This study investigates in vivo mechanism action of SOL treatment in L. amazonensis-infected BALB/c mice. SOL was isolated from the roots of S. chilensis, and L. amazonensis-infected mice were treated daily with SOL (10, 50, 100 mg/kg) by gavage for 30 days. Gastric (NAG, MPO), hepatic (AST, ALT), systemic (body weight, NO) toxicity, leishmanicidal activity (lesion size, parasite burden), cell profile (macrophage, neutrophil infiltration), antioxidant (ABTS, NBT, NO), oxidant parameters (FRAP, ABTS), Th1, Th2, Th17 cytokines (CBA), collagen deposition (picrosirius), arginase, iNOS, NF-kB, and NRF2 (immunofluorescence) were evaluated. In vivo results showed SOL-treatment did not induce gastric, hepatic, or systemic toxicity in L. amazonensis-infected mice. SOL was able to reduce the lesion size and parasite load at the site of infection, increasing macrophage infiltration and neutrophil migration, exerting a balance in antioxidant (increased ABTS, NBT reduction, and NO), oxidative (increased FRAP and ABTS), and anti-inflammatory responses (reduced TNF-α, IFN-γ and increased IL-6, IL-17 production), and inducing arginase, iNOS, NF-kB, NRF2 and collagen deposition (type III), favoring wound healing and accelerating tissue repair at the site injury.

Spray-drying of casein/pectin bioconjugate microcapsules containing grape (Vitis labrusca) by-product extract.

Novel microcapsules containing grape peel by-product extract were obtained. In this pursuit, complex coacervation of casein/pectin bioconjugate and spray-drying were combined. We have investigated the role of the dispersion feed rate (FR), drying air inlet temperature (IT) and drying air flow rate (AR) in the drying yield, microencapsulation efficiency, total polyphenols and anthocyanins contents, antioxidant activity, and morphology of the products. Also, the first-order degradation kinetics of the phytochemicals for both the extract and dried microcapsules was assessed and compared. The loss on the phytochemicals during spray-drying was attenuated in up to 88%, and the IT was the main factor affecting the particle properties. The polyphenols on the extract interacted with the polymers, influencing the assemble of the bioconjugate and the particle's features. Such microencapsulation strategy enhanced the thermal stability of the phytochemicals and rendered biocompatible and biodegradable products of which the nutraceutical and cosmeceutical application may have potential.

Grandiflorenic acid isolated from Sphagneticola trilobata against Trypanosoma cruzi: Toxicity, mechanisms of action and immunomodulation.

Grandiflorenic acid (GFA) is one of the main kaurane diterpenes found in different parts of Sphagneticola trilobata. It has several biological activities, especially antiprotozoal action. In turn, Chagas disease is a complex systemic disease caused by the protozoan Trypanosoma cruzi, and the drugs available to treat it involve significant side effects and impose an urgent need to search for therapeutic alternatives. In this context, our goal was to determine the effect of GFA on trypomastigote and intracellular amastigote forms. Our results showed that GFA treatment led to significantly less viability of trypomastigote forms, with morphological and ultrastructural changes in the parasites treated with IC50 of GFA (24.60 nM), and larger levels of reactive oxygen species (ROS), mitochondrial depolarization, lipid droplets accumulation, presence of autophagic vacuoles, phosphatidylserine exposure, and plasma membrane damage. In addition, the GFA treatment was able to reduce the percentage of infected cells and the number of amastigotes per macrophage (J774A.1) without showing cytotoxicity in mammalian cell lines (J774A.1, LLCMK2, THP-1, AMJ2-C11), in addition to increasing TNF-α and reducing IL-6 levels in infected macrophages. In conclusion, the GFA treatment exerted influence on trypomastigote forms through an apoptosis-like mechanism and by eliminating intracellular parasites via TNF-α/ROS pathway, without generating cellular cytotoxicity.

Recent Advances in Biotransformation by Cunninghamella Species.

The goal of the biotransformation process is to develop structural changes and generate new chemical compounds, which can occur naturally in mammalian and microbial organisms, such as filamentous fungi, and represent a tool to achieve enhanced bioactive compounds. Cunninghamella spp. is among the fungal models most widely used in biotransformation processes at phase I and II reactions, mimicking the metabolism of drugs and xenobiotics in mammals and generating new molecules based on substances of natural and synthetic origin. Therefore, the goal of this review is to highlight the studies involving the biotransformation of Cunninghamella species between January 2015 and March 2021, in addition to updating existing studies to identify the similarities between the human metabolite and Cunninghamella patterns of active compounds, with related advantages and challenges, and providing new tools for further studies in this scope.

Solidagenone acts on promastigotes of L. amazonensis by inducing apoptosis-like processes on intracellular amastigotes by IL-12p70/ROS/NO pathway activation.

BACKGROUND: Leishmaniasis is a neglected tropical disease caused by protozoan parasites of the Leishmania genus. Currently, the treatment has limited effectiveness and high toxicity, is expensive, requires long-term treatment, induces significant side effects, and promotes drug resistance. Thus, new therapeutic strategies must be developed to find alternative compounds with high efficiency and low cost. Solidagenone (SOL), one of the main constituents of Solidago chilensis, has shown gastroprotective, anti-inflammatory and immunomodulatory effects. PURPOSE: This study assessed the in vitro effect of SOL on promastigotes and Leishmania amazonensis-infected macrophages, as well its microbicide and immunomodulatory mechanisms. METHODS: SOL was isolated from the roots of S. chilensis, 98% purity, and identified by chromatographic methods, and the effect of SOL on leishmanicidal activity against promastigotes in vitro, SOL-induced cytotoxicity in THP-1, J774 cells, sheep erythrocytes, and L. amazonensis-infected J774 macrophages, and the mechanisms of death involved in this action were evaluated. RESULTS: In silico predictions showed good drug-likeness potential for SOL with high oral bioavailability and intestinal absorption. SOL treatment (10-160 µM) inhibited promastigote proliferation 24, 48, and 72 h after treatment. After 24 h of treatment, SOL at the IC50 (34.5 µM) and 2 × the IC50 (69 µM) induced several morphological and ultrastructural changes in promastigotes, altered the cell cycle and cellular volume, increased phosphatidylserine exposure on the cell surface, induced the loss of plasma membrane integrity, increased the reactive oxygen species (ROS) level, induced loss of mitochondrial integrity (characterized by an apoptosis-like process), and increased the number of lipid droplets and autophagic vacuoles. Additionally, SOL induced low cytotoxicity in J774 murine macrophages (CC50 of 1587 µM), THP-1 human monocytes (CC50 of 1321 µM), and sheep erythrocytes. SOL treatment reduced the percentage of L. amazonensis-infected macrophages and the number of amastigotes per macrophage (IC50 9.5 µM), reduced TNF-α production and increased IL-12p70, ROS and nitric oxide (NO) levels. CONCLUSION: SOL showed in vitro leishmanicidal effects against the promastigotes by apoptosis-like mechanism and amastigotes by reducing TNF-α and increasing IL-12p70, ROS, and NO levels, suggesting their potential as a candidate for use in further studies on the design of antileishmanial drugs.

Renicolidae infection in Manx shearwater (Puffinus puffinus): is parasitism implicated on renal lesions?

Renicolid digeneans are frequently observed in the renal tubules and ureters of seabirds, such Puffinus puffinus, a migratory species distributed along the Brazilian coast. However, few studies have focused on the relationship between renicolid infection and health status in P. puffinus. Thus, the aim of this study was to describe (i) renal and systemic alterations, (ii) the renicolids and (iii) the biological aspects associated with the presence of renicolids in P. puffinus. Gross and histological assays were performed in 93 P. puffinus stranded on the Paraná coast, southern Brazil, and renicolids were submitted to morphological and molecular assays. A high prevalence of renicolids in P. puffinus (71/93) was observed. In the kidney, the main microscopic findings were lymphocytic interstitial infiltrate, ductal ectasia and tubular necrosis. The renal lesions were significantly associated with the parasite infection. The morphological (n = 84) and molecular analyses (n = 2) confirmed the species as Renicola sloanei (100% and 95.9% of nucleotide identity with R. sloanei strains from P. puffinus and from Spheniscus demersus, respectively). In both parasitized and non-parasitized animals, cardiac and skeletal muscle degeneration and necrosis were the most frequent systemic changes. Therefore, the results suggest renicolids being a possible cause for the demonstrated renal alterations. A contribution of this parasite to a decreased health status of Puffinus puffinus along their migratory route is possible.

Seroepidemiology of ovine toxoplasmosis and neosporosis in breeding rams from Rio Grande do Sul, Brazil.

This study determined the prevalence of ovine toxoplasmosis and neosporosis and the risk factors associated with the development of these diseases in breeding rams from the State of Rio Grande do Sul (RS), Southern Brazil. Serum samples (n = 1,800) from breeding rams maintained on 705 sheep farms from seven mesoregions were evaluated serologically to detect anti-IgG Toxoplasma gondii by indirect ELISA and anti-IgG Neospora caninum by the indirect immunofluorescence assay (IFA). The prevalence of T. gondii was 33.05% (595/1,800); seropositivity to N. caninum was 18.44% (332/1,800). Additionally, there was simultaneous seropositivity (8.94%;161/1,800) to N. caninum and T. gondii. The variables size of the property (<500 ha) (Prevalence Ratio, PR = 1.36); breeding system (semi-intensive/intensive) (PR = 1.23); and natural mounting without control (PR = 1.50) were considered as risk factors for the occurrence of T. gondii. Size of the property (<500 ha) (PR = 1.58) and natural mounting without control (PR = 2.32) were risk factors associated with the prevalence of N. caninum in rams. Additionally, separation of ewes prior to parturition was considered as a protective factor for the occurrence of T. gondii (PR = 0.82) and N. caninum (PR = 0.74). These results demonstrated that these two parasitic disease agents are endemic in rams throughout all regions of RS.

Epidemiology of a toxoplasmosis outbreak in a research institution in northern Paraná, Brazil.

Toxoplasmosis is a reportable disease in Brazil. The objective of this study was to investigate a toxoplasmosis outbreak at a research institution in Londrina-PR, Brazil. The outbreak was reported in October 2015; however, the first cases occurred in August 2015. Blood samples were collected from 674 persons at the institution. Samples were collected from soil, water (water tank) and food (vegetables) served in the restaurant. Each participant responded to an epidemiological questionnaire. For the blood samples, a chemiluminescent microparticle immunoassay was performed to detect IgM, IgG and specific IgG avidity antibodies; 10.8% (73/674) had evidence of acute toxoplasmosis. Statistical analysis showed a significant association (p < .001) between acute infection and eating lunch in the restaurant of the institution. Regarding the types of food offered in the restaurant during the period, there was a significant association between consuming raw salad (p < .001) and becoming ill. We conclude that the vegetables or raw vegetables served in the restaurant were probably the source of infection; however, the long period between exposure and case reporting made it difficult to identify the source of transmission.

Isolamento do estigmasterol do extrato hexânico das folhas de Sphagneticola trilobata Pruski (Asteraceae) / Isolation of stigmaterol from hexanic extract of the leaves of Sphagneticola trilobata Pruski (Asteraceae)

O interesse de pesquisadores e da população geral é bastante evidente, e tem crescido bastante na atualidade, no uso de plantas medicinais. A espécie de interesse deste trabalho, a Sphagneticola trilobata Pruski é uma planta herbácea pertencente à família Asteraceae. Alguns relatos têm mostrado a presença de diversos constituintes químicos nesta planta, principalmente diterpenos. O presente trabalho realizou o estudo fitoquímico de uma fração do extrato hexânico, devido ao baixo rendimento da fração diclorometânica, através da utilização de diversos métodos cromatográficos, na qual isolouse o fitoesterol estigmasterol. Este composto foi confirmado por meio de dados espectroscópicos e de literaturas, e o isolamento de compostos desta classe de substâncias indicam uma nova vertente para o uso medicinal desta planta como um futuro hipocolesterolêmico. Para isto, estudos serão direcionados para o isolamento de fitoesteróis, com posterior realização de ensaios biológicos (AU)

The interest of researchers and the general population is quite evident, and has grown considerably today, in the use of medicinal plants. The species of interest in this work, Sphagneticola trilobata Pruski is a herbaceous plant, belonging to Asteraceae family. Some reports have shown the presence of several chemical constituents in this plant, mainly diterpenes. The present work carried out the phytochemical study of a fraction of the hexane extract, due to the low yield of the dichloromethane fraction, by several chromatographic methods, in which the phytosterol estigmasterol was isolated. This compound has been confirmed by means of spectroscopic and literature data, and the isolation of compounds of this class of substances indicates a new strand for the medicinal use of this plant as a hypocholesterolemic future. For this, studies will be directed to the isolation of phytosterols, with subsequent biological tests (AU)

Investigation and environmental analysis of samples from outbreak of toxoplasmosis at research institution in Londrina, Paraná, Brazil, 2016.

The objective of this study was to report an outbreak of human toxoplasmosis at a research institution in Londrina, Paraná, from December 2015 to February 2016. Blood samples from 26 symptomatic individuals were collected and the microparticle chemiluminescence immunoassay was performed to detect IgM, IgG and specific IgG avidity test in the official laboratory. A total of 20 people with symptoms and serology compatible with acute toxoplasmosis (IgM positive and IgG with low avidity) were selected as cases, while 45 asymptomatic employees working in the same teams and during the same shifts were selected as controls. All the participants of the investigation answered an epidemiological questionnaire. Three samples of water and one sludge from the institution's supply cisterns, 10 soil samples, 11 plant samples, three cat fecal samples and one domestic feline cadaver were collected for analysis of the polymerase chain reaction (PCR) for T. gondii. After analyzing the epidemiological data, the consumption of vegetables in the restaurant of the institution was the only variable associated with the occurrence of the disease. In laboratory results, all the samples showed negative results to PCR. The rapid recognition of the outbreak, early notification and investigation could have broken the chain of transmission early, thus preventing the emergence of new cases. In addition, the adoption of good food handling practices could have prevented the occurrence of the outbreak.

Investigation and environmental analysis of samples from outbreak of toxoplasmosis at research institution in Londrina, Paraná, Brazil, 2016 / Investigação e análise ambiental de amostras oriundas de surto de toxoplasmose em uma instituição de pesquisa em Londrina, Paraná, Brasil, 2016

Abstract The objective of this study was to report an outbreak of human toxoplasmosis at a research institution in Londrina, Paraná, from December 2015 to February 2016. Blood samples from 26 symptomatic individuals were collected and the microparticle chemiluminescence immunoassay was performed to detect IgM, IgG and specific IgG avidity test in the official laboratory. A total of 20 people with symptoms and serology compatible with acute toxoplasmosis (IgM positive and IgG with low avidity) were selected as cases, while 45 asymptomatic employees working in the same teams and during the same shifts were selected as controls. All the participants of the investigation answered an epidemiological questionnaire. Three samples of water and one sludge from the institution's supply cisterns, 10 soil samples, 11 plant samples, three cat fecal samples and one domestic feline cadaver were collected for analysis of the polymerase chain reaction (PCR) for T. gondii. After analyzing the epidemiological data, the consumption of vegetables in the restaurant of the institution was the only variable associated with the occurrence of the disease. In laboratory results, all the samples showed negative results to PCR. The rapid recognition of the outbreak, early notification and investigation could have broken the chain of transmission early, thus preventing the emergence of new cases. In addition, the adoption of good food handling practices could have prevented the occurrence of the outbreak.

Resumo O objetivo deste estudo foi relatar um surto de toxoplasmose humana em uma instituição de pesquisa em Londrina, Paraná, no período de dezembro de 2015 a fevereiro de 2016. Amostras de sangue de 26 indivíduos sintomáticos foram coletadas e o imunoensaio de quimioluminescência de micropartículas foi realizado para detectar IgM, IgG e teste de avidez de IgG específica em laboratório oficial. Um total de 20 pessoas com sintomas e sorologia compatíveis com toxoplasmose aguda (IgM positiva e IgG com baixa avidez) foi selecionado como casos, enquanto 45 funcionários assintomáticos que trabalhavam nas mesmas equipes e durante os mesmos turnos foram utilizados como controles. Todos os participantes da investigação responderam a um questionário epidemiológico. Foram coletadas três amostras de água e uma de lodo das cisternas de abastecimento da instituição, 10 de solo, 11 de vegetais, três amostras de fezes de gato e um cadáver de filhote felino doméstico para detecção de T. gondii pela reação em cadeia da polimerase (PCR). Após análise dos dados epidemiológicos, o consumo de hortaliças no restaurante da instituição foi a única variável associada à ocorrência da doença. Em resultados laboratoriais, todas as amostras apresentaram resultados negativos a PCR. O rápido reconhecimento do surto, notificação e investigação prematura poderia ter quebrado a cadeia de transmissão, evitando assim o surgimento de novos casos. Além disso, a adoção de boas práticas de manipulação de alimentos poderia ter impedido a ocorrência do surto.