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1.
Mol Divers ; 26(4): 1969-1982, 2022 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-34482477

RESUMO

Giardiasis is a neglected disease, and there is a need for new molecules with less side effects and better activity against resistant strains. This work describes the evaluation of the giardicidal activity of thymol derivatives produced from the Morita-Baylis-Hillman reaction. Thymol acrylate was reacted with different aromatic aldehydes, using 1,4-diazabicyclo[2.2.2]octane (DABCO) as a catalyst. Eleven adducts (8 of them unpublished) with yields between 58 and 80% were obtained from this reaction, which were adequately characterized. The in silico prediction showed theoretical bioavailability after oral administration as well as antiparasitic activity against Giardia lamblia. Compound 4 showed better biological activity against G. lamblia. In addition to presenting antigiardial activity 24 times better than thymol, this MBHA was obtained in a short reaction time (3 h) with a yield (80%) superior to the other investigated molecules. The molecule was more active than the precursors (thymol and MBHA 12) and did not show cytotoxicity against HEK-293 or HT-29 cells. In conclusion, this study presents a new class of drugs with better antigiardial activity in relation to thymol, acting as a basis for the synthesis of new bioactive molecules. Molecular hybridization technique combined with the Morita-Baylis-Hillman reaction provided new thymol derivatives with giardicidal activity superior to the precursor molecules.


Assuntos
Giardia lamblia , Timol , Aldeídos , Catálise , Células HEK293 , Humanos , Timol/farmacologia
2.
Exp Parasitol ; 230: 108158, 2021 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-34534534

RESUMO

Our aim was to evaluate the impact of immunosuppression on the development of giardiasis. Thirty-six gerbils (4-6 weeks old) were distributed in four groups containing nine animals each: Control (CT); Control-Infected by Giardia lamblia (CTIn), Immunosuppressed (IS), and Immunosuppressed-Infected by G. lamblia (ISIn). Animals in the IS and ISIn groups received intramuscular dexamethasone solution for 25 days. On the 11th day, the animals in the CTIn and ISIn groups were inoculated with G. lamblia. After 14 days of infection, the 25th day of the experiment, all groups were euthanized. Four hours after euthanasia, the intestinal permeability was evaluated and sections of the duodenum and spleen were harvested for morphometric and histopathological analyses. Immunosuppressed groups showed a significant increase in intestinal permeability compared to control and infected groups. Considering that the infection can become chronic in immunosuppressed groups, we should be alert to the possibilities of chronic inflammatory changes, both locally and systemically, due to the loss of the intestinal barrier. Lesions were observed in the duodenal mucosa of the gerbils of the CTIn group, with reduced villi size, crypt hyperplasia, edema, and the presence of inflammatory infiltrate in the lamina propria. In the ISIn group, we observed no inflammation, long and intact villi, and a significant increase in the area of intestinal mucins, despite the large number of trophozoites identified. Our results suggest that exacerbation of the immune response has a direct relationship with the appearance of lesions during enteritis produced by G. lamblia in the assessed model.


Assuntos
Dexametasona/uso terapêutico , Enterite/tratamento farmacológico , Enterite/parasitologia , Giardíase/tratamento farmacológico , Glucocorticoides/uso terapêutico , Animais , Dexametasona/farmacologia , Modelos Animais de Doenças , Duodeno/parasitologia , Duodeno/patologia , Enterite/imunologia , Feminino , Gerbillinae , Giardia lamblia/efeitos dos fármacos , Giardia lamblia/imunologia , Giardia lamblia/patogenicidade , Giardíase/imunologia , Giardíase/parasitologia , Glucocorticoides/farmacologia , Terapia de Imunossupressão , Mucosa Intestinal/parasitologia , Mucosa Intestinal/patologia , Masculino , Carga Parasitária , Permeabilidade , Baço/patologia
3.
ScientificWorldJournal ; 2014: 364264, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-24605052

RESUMO

Entamoeba histolytica is a protozoan parasite that presents a risk to the health of millions of people worldwide. Due to the existence of different clinical forms caused by the parasite and also different virulence levels presented by one strain, one would expect differences in the profile of gene transcripts between virulent and nonvirulent cultures. In this study we used the differential display to select gene segments related to invasiveness of amoeba. One Brazilian strain of E. histolytica in two conditions, able or not to cause lesions in experimental animals, was used. RNA from this strain, was used to study the differential expression of genes. 29 specific gene fragments differentially expressed in the virulent strain were selected. By real-time PCR, six of these genes had confirmed their differential expression in the virulent culture. These genes may have important roles in triggering invasive amoebiasis and may be related to adaptation of trophozoites to difficulties encountered during colonization of the intestinal epithelium and liver tissue. Future studies with these genes may elucidate its actual role in tissue invasion by E. histolytica generating new pathways for diagnosis and treatment of amoebiasis.


Assuntos
Entamoeba histolytica/metabolismo , Entamebíase/metabolismo , Regulação da Expressão Gênica , RNA de Protozoário/biossíntese , Animais , Entamoeba histolytica/genética , Entamoeba histolytica/patogenicidade , Entamebíase/genética , Entamebíase/terapia , Humanos , Camundongos , RNA de Protozoário/genética , Ratos
4.
Rev Panam Salud Publica ; 36(1): 37-43, 2014 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-25211676

RESUMO

OBJECTIVE: To assess the additional cost of incorporating the detection and treatment of retinopathy of prematurity (ROP) into neonatal care services of Brazil's Unified Health System (SUS). METHODS: A deterministic decision-tree simulation model was built to estimate the direct costs of screening for and treating ROP in neonatal intensive-care units (NICUs), based on data for 869 preterm infants with birth weight less than 1 500 g examined in six governmental NICUs in the capital city of Rio de Janeiro, where coverage was 52% and 8% of infants were treated. All of the parameters from this study were extrapolated to Brazilian newborn estimates in 2010. Costs of screening and treatment were estimated considering staff, equipment and maintenance, and training based on published data and expert opinion. A budget impact analysis was performed considering the population of preterm newborns, screening coverage, and the incidence of treatable ROP. One- and two-way sensitivity analyses were performed. RESULTS: In Rio de Janeiro, unit costs per newborn were US$ 18 for each examination, US$ 398 per treatment, and US$ 29 for training. The estimated cost of ROP diagnosis and treatment for all at-risk infants NICUs was US$ 80 per infant. The additional cost to the SUS for one year would be US$ 556 640 for a ROP program with 52% coverage, increasing to US$ 856 320 for 80% coverage, and US$ 1.07 million or 100% coverage. CONCLUSIONS: The results of this study indicate that providing ROP care is affordable within the framework of the SUS in Brazil, and might be feasible elsewhere in Latin America, considering the evidence of the effectiveness of ROP treatment and the social benefits achieved.


Assuntos
Custos de Cuidados de Saúde , Triagem Neonatal/economia , Retinopatia da Prematuridade/diagnóstico , Retinopatia da Prematuridade/terapia , Brasil , Árvores de Decisões , Humanos , Recém-Nascido , Unidades de Terapia Intensiva Neonatal , Retinopatia da Prematuridade/economia
5.
Biomedicines ; 12(3)2024 Mar 07.
Artigo em Inglês | MEDLINE | ID: mdl-38540211

RESUMO

Complex structural chromosome abnormalities such as chromoanagenesis have been reported in acute myeloid leukemia (AML). They are usually not well characterized by conventional genetic methods, and the characterization of chromoanagenesis structural abnormalities from short-read sequencing still presents challenges. Here, we characterized complex structural abnormalities involving chromosomes 2, 3, and 7 in an AML patient using an integrated approach including CRISPR/Cas9-mediated nanopore sequencing, mate pair sequencing (MPseq), and SNP microarray analysis along with cytogenetic methods. SNP microarray analysis revealed chromoanagenesis involving chromosomes 3 and 7, and a pseudotricentric chromosome 7 was revealed by cytogenetic methods. MPseq revealed 138 structural variants (SVs) as putative junctions of complex rearrangements involving chromosomes 2, 3, and 7, which led to 16 novel gene fusions and 33 truncated genes. Thirty CRISPR RNA (crRNA) sequences were designed to map 29 SVs, of which 27 (93.1%) were on-target based on CRISPR/Cas9 crRNA nanopore sequencing. In addition to simple SVs, complex SVs involving over two breakpoints were also revealed. Twenty-one SVs (77.8% of the on-target SVs) were also revealed by MPseq with shared SV breakpoints. Approximately three-quarters of breakpoints were located within genes, especially intronic regions, and one-quarter of breakpoints were intergenic. Alu and LINE repeat elements were frequent among breakpoints. Amplification of the chromosome 7 centromere was also detected by nanopore sequencing. Given the high amplification of the chromosome 7 centromere, extra chromosome 7 centromere sequences (tricentric), and more gains than losses of genomic material, chromoanasynthesis and chromothripsis may be responsible for forming this highly complex structural abnormality. We showed this combination approach's value in characterizing complex structural abnormalities for clinical and research applications. Characterization of these complex structural chromosome abnormalities not only will help understand the molecular mechanisms responsible for the process of chromoanagenesis, but also may identify specific molecular targets and their impact on therapy and overall survival.

6.
Exp Parasitol ; 133(4): 391-5, 2013 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-23337825

RESUMO

Giardiasis is one of the most common parasitic diseases worldwide, and the disease is an important cause of diarrhoea and malabsorption in children and immunosuppressed individuals. However, there is no evidence that characterises malnutrition as an aggravating factor for this disease. We evaluated changes in villi structures to examine the association between malnutrition and Giardia lamblia infection. We used 32 gerbils, divided into 4 groups: Control (CT) and Control Infected (CTIn), which each received a 20% protein diet, Malnourished (MN) and Malnourished Infected (MNIn), which each received a 5% protein diet. Groups CTIn and MNIn were inoculated with 1×10(6) trophozoites of G. lamblia, while the remaining groups were mock infected. Seven days post-infection, all groups were sacrificed, and the proximal portions of the small intestines were collected for the analysis of villus height, mucus area and extent of Giardia infection. Gerbils fed with a low-protein diet had significantly lower body weights. Malnourished infected animals presented significantly increased production of mucus, suggesting a synergism occurs between malnutrition and Giardiasis, potentially to control the adhesion of Giardia in the mucosa. Villus height was significantly lower in group MNIn compared to CTIn. This work suggests that malnutrition contributes to severity of Giardiasis by decreasing the intestinal absorption capacity via shortening of the villi.


Assuntos
Giardíase/complicações , Giardíase/patologia , Intestino Delgado/patologia , Desnutrição Proteico-Calórica/complicações , Desnutrição Proteico-Calórica/patologia , Animais , Feminino , Gerbillinae , Células Caliciformes/metabolismo , Células Caliciformes/patologia , Mucosa Intestinal/metabolismo , Mucosa Intestinal/parasitologia , Mucosa Intestinal/patologia , Intestino Delgado/metabolismo , Intestino Delgado/parasitologia , Microvilosidades/metabolismo , Microvilosidades/parasitologia , Microvilosidades/patologia , Muco/metabolismo
7.
PLoS Negl Trop Dis ; 16(5): e0010105, 2022 05.
Artigo em Inglês | MEDLINE | ID: mdl-35499991

RESUMO

BACKGROUND: Epidemiological and experimental studies have shown a protective effect of helminth infections in weight gain and against the development of metabolic dysfunctions in the host. However, the mechanisms Treg cells exert in the helminth-obesity interface has been poorly investigated. The present study aimed to verify the influence of Heligmosomoides polygyrus infection in early stages of high fat diet-induced obesity. PRINCIPAL FINDINGS: The presence of infection was able to prevent exacerbated weight gain in mice fed with high fat diet when compared to non-infected controls. In addition, infected animals displayed improved insulin sensitivity and decreased fat accumulation in the liver. Obesity-associated inflammation was reduced in the presence of infection, demonstrated by lower levels of leptin and resistin, lower infiltration of Th1 and Th17 cells in adipose tissue, higher expression of IL10 and adiponectin, increased infiltration of Th2 and eosinophils in adipose tissue of infected animals. Of note, the parasite infection was associated with increased Treg frequency in adipose tissue which showed higher expression of cell surface markers of function and activation, like LAP and CD134. The infection could also increase adipose Treg suppressor function in animals on high fat diet. CONCLUSION: These data suggest that H. polygyrus modulates adipose tissue Treg cells with implication for weight gain and metabolic syndrome.


Assuntos
Dieta Hiperlipídica , Resistência à Insulina , Tecido Adiposo , Animais , Dieta Hiperlipídica/efeitos adversos , Resistência à Insulina/fisiologia , Camundongos , Camundongos Endogâmicos C57BL , Obesidade/metabolismo , Aumento de Peso
8.
Exp Parasitol ; 122(4): 306-9, 2009 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-19389401

RESUMO

Cysteine proteinase (CP) activity and CP5 mRNA levels were analyzed in eleven samples of Entamoeba histolytica isolated from patients presenting different clinical profiles. The virulence degree of the isolates, determined in hamster liver, correlated well with the clinical form of the patient and culture conditions. CP5 mRNA levels were also determined in sample freshly picked up directly from liver amoebic abscess. Differences were not observed in the levels of CP5 mRNA and CP specific activity among the cultured samples. However, different levels of CP5 mRNA were observed in trophozoite freshly isolated from hepatic amoebic lesions. These results reinforce the importance of CP5 for the virulence of amoebae and the need for studies with the parasite present in lesions to validate mechanisms involved in pathogenesis of amoebiasis.


Assuntos
Cisteína Endopeptidases/metabolismo , Entamoeba histolytica/enzimologia , Entamoeba histolytica/patogenicidade , Entamebíase/parasitologia , Abscesso Hepático Amebiano/parasitologia , Animais , Cricetinae , Cisteína Endopeptidases/genética , Entamoeba histolytica/genética , Entamebíase/patologia , Regulação Enzimológica da Expressão Gênica , Humanos , Fígado/parasitologia , Fígado/patologia , Abscesso Hepático Amebiano/patologia , Mesocricetus , Reação em Cadeia da Polimerase , RNA Mensageiro/análise , Virulência
9.
Acta Trop ; 190: 112-118, 2019 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-30447179

RESUMO

Trichomonas vaginalis is an amitochondrial parasite that causes human trichomoniasis. Despite metronidazole effectiveness, resistant cases are becoming more frequent. This scenario reveals the need to develop new therapeutic options. Photodynamic Therapy (PDT) is an experimental treatment that involves the activation of photosensitive substances and the generation of cytotoxic oxygen species and free radicals to promote the selective destruction of target tissues. In previous work, we identified an excellent in vitro PDT activity using methylene blue and light emitting diode against metronidazole sensitive and resistant strains of T. vaginalis. Here, we evaluated the efficacy of PDT in vivo and its high trichomonicidal activity was assessed through transmission electron microscopy. Female Balb/c mice were infected intravaginally with T. vaginalis trophozoites. On the third day of infection, methylene blue was introduced into the vaginal canal, which then received 68.1 J/cm2 of radiation for 35.6 s. Twenty-four hours after treatment the vaginal canal of the animals was scraped and the samples processed by the immunocytochemistry technique. Besides that, in vitro photodynamic treatment was performed and T. vaginalis trophozoites were processed by transmission electron microscopy. PDT significantly reduced infection in animals treated, compared to control groups, being as efficient as metronidazole. Morphological changes observed have suggested that PDT activity on T. vaginalis was due to necrosis. These results, added to the high trichomonicidal activity of PDT confirm its feasibility for trichomoniasis treatment.


Assuntos
Microscopia Eletrônica de Transmissão/métodos , Fotoquimioterapia , Vaginite por Trichomonas/tratamento farmacológico , Trichomonas vaginalis/efeitos dos fármacos , Animais , Feminino , Humanos , Metronidazol/uso terapêutico , Camundongos , Camundongos Endogâmicos BALB C , Trichomonas vaginalis/ultraestrutura
10.
Bioorg Med Chem ; 16(2): 854-61, 2008 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-17977731

RESUMO

This study evaluated the effects of flavone eupafolin (6-methoxy 5,7,3',4'-tetrahydroxyflavone), extracted from dry leaves of Eupatorium litoralle. Eupafolin (25-200microM) promoted inhibition of the respiratory rate in state 3, in the presence of glutamate or succinate. During succinate oxidation, it was found that only state 4 respiratory rate was stimulated approximately 30% by eupafolin (100microM) and ADP/O ratio and RCC were reduced with all doses. When glutamate was used as substrate, RCC was similarly reduced. Eupafolin caused a reduction of enzymatic activities between complexes I and III of the respiratory chain. Cytochrome c oxidase and ATPase activities were not affected. Using voltammetry cyclic analysis, eupafolin give rise to irreversible oxidation with an anodic peak potential at +0.08V (SHE). We also observed that eupafolin can undergo oxidation catalyzed by EDTA-Fe, promoting cytochrome c reduction in the presence of NADH, resulting in the production of the superoxide radical and hydrogen peroxide. All together, the results could explain the cytotoxic effects observed previously with the eupafolin.


Assuntos
Respiração Celular/efeitos dos fármacos , Flavonas/farmacologia , Mitocôndrias/efeitos dos fármacos , Adenosina Trifosfatases/efeitos dos fármacos , Complexo IV da Cadeia de Transporte de Elétrons/efeitos dos fármacos , Flavonas/química , Flavonas/isolamento & purificação , Flavonas/toxicidade , Ácido Glutâmico/metabolismo , Mitocôndrias/metabolismo , Estrutura Molecular , Oxirredução , Relação Estrutura-Atividade
11.
BMC Genomics ; 8: 170, 2007 Jun 14.
Artigo em Inglês | MEDLINE | ID: mdl-17567921

RESUMO

BACKGROUND: A number of studies have shown that peptidases and in particular cysteine peptidases constitute major pathogenicity factors in Entamoeba histolytica. Recent studies have suggested that a considerable number of genes coding for proteolytic enzymes are present within the E. histolytica genome and questions remain about the mode of expression of the various molecules. RESULTS: By homology search within the recently published amoeba genome, we identified a total of 86 E. histolytica genes coding for putative peptidases, including 46 recently described peptidase genes. In total these comprise (i) 50 cysteine peptidases of different families but most of which belong to the C1 papain superfamily, (ii) 22 different metallo peptidases from at least 11 different families, (iii) 10 serine peptidases belonging to 3 different families, and (iv) 4 aspartic peptidases of only one family. Using an oligonucleotide microarray, peptidase gene expression patterns of 7 different E. histolytica isolates as well as of heat stressed cells were analysed. A total of 21 out of 79 amoeba peptidase genes analysed were found to be significantly expressed under standard axenic culture conditions whereas the remaining are not expressed or at very low levels only. In heat-stressed cells the expression of 2 and 3 peptidase genes, respectively, were either decreased or increased. Only minor differences were observed between the various isolates investigated, despite the fact that these isolates were originated from asymptomatic individuals or from patients with various forms of amoebic diseases. CONCLUSION: Entamoeba histolytica possesses a large number of genes coding for proteolytic enzymes. Under standard culture conditions or upon heat-stress only a relatively small number of these genes is significantly expressed and only very few variations become apparent between various clinical E. histolytica isolates, calling into question the importance of these enzymes in E. histolytica pathogenicity. Further studies are required to define the precise role of most of the proteolytic enzyme for amoeba cell biology but in particular for E. histolytica virulence.


Assuntos
Entamoeba histolytica/genética , Genoma de Protozoário/genética , Peptídeo Hidrolases/genética , Animais , Sequência de Bases , Biologia Computacional , Bases de Dados de Ácidos Nucleicos , Perfilação da Expressão Gênica , Temperatura Alta , Análise de Sequência com Séries de Oligonucleotídeos , Peptídeo Hidrolases/química
12.
Parasitol Res ; 102(1): 145-9, 2007 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-17906962

RESUMO

We comparatively evaluate the effect of metronidazole (MTZ) and its five analogues on trophozoites of Giardia lamblia axenically growing. The compounds MTZ-Ms, MTZ-I, MTZ-Br, MTZ-N(3), and MTZ-NH(3)Cl were obtained by molecular modification of the side chain of MTZ. Four of them presented higher giardicidal activity when compared with MTZ. Among them, MTZ-Br and MTZ-I were the most active, without cytotoxic effects against mitogen-activated human peripheral blood mononuclear cells (PBMC). The alteration of MTZ side chain constitutes a fruitful field to develop new drugs for the treatment not only of giardiasis but also of other diseases and signalize that metronidazole analogues are promising candidates as giardicidal and should be further evaluated.


Assuntos
Antiprotozoários/farmacologia , Giardia lamblia/efeitos dos fármacos , Metronidazol/análogos & derivados , Metronidazol/farmacologia , Animais , Células Cultivadas , Relação Dose-Resposta a Droga , Giardia lamblia/citologia , Humanos , Concentração Inibidora 50 , Leucócitos Mononucleares/efeitos dos fármacos , Metronidazol/química , Estereoisomerismo , Relação Estrutura-Atividade
13.
PeerJ ; 5: e2967, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28265495

RESUMO

BACKGROUND: Nowadays, the number of obese people in the world has reached alarming proportions. During the expansion of adipose tissue, a number of functions such as activation and release of cytokines and hormones may be affected. This leads the body to a pro-inflammatory pattern, which may affect the proper functioning of many tissues. Thus, studying the mechanisms by which obesity induces physiological disorders is necessary, and may be facilitated by the use of animal models, in particular rodents. We sought to characterize the metabolic and adipose tissue changes resulting from a diet rich in fats and simple sugars in gerbils. METHODS: We divided 14 gerbils into two experimental groups that received a diet rich in simple carbohydrates and fats with 5,86 kcal/g (OB, n = 7) or a standard diet with 4.15 kcal/g (CT; n = 7) for 11 weeks. The animals had free access to water and food. The animal weight and food consumption were measured weekly. Blood, adipose tissue and liver of each animal were collected at the end of experiment. The following parameters were determined: cholesterol (COL), triglycerides (TGL) and glycemia (GLI) in the plasma; cytokines (IL-6, IL-10 and TNF-α) and hormones (adiponectin and leptin) in adipose tissue; activity of superoxide dismutase (SOD) and catalase (CAT), extraction and differentiation of fat and histology in liver. RESULTS: The consumption of a diet rich in simple carbohydrates and fats led to increased total body weight and increased relative weights of liver and adipose tissue. In addition, we observed increased fasting glucose levels and circulating triglycerides, along with high TNF-α production in adipose tissue and increased total fat, cholesterol and triglyceride contents in the liver, contributing to higher intensity of hepatic steatosis. On the other hand, the animals of this group showed depletion in the enzyme activity of SOD and CAT in the liver, as well as reduction of IL-10 and adiponectin levels in adipose tissue. DISCUSSION: High intake of saturated fat and simple carbohydrates establish the gerbil as an experimental model for the study of metabolic and hepatic abnormalities resulting from obesity.

14.
Mol Biochem Parasitol ; 203(1-2): 34-8, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26542948

RESUMO

Leishmania major, the causative agent of zoonotic leishmaniasis, is restricted to Old World countries. Molecular and biochemical techniques have been used to identify some L. major-like isolated in South America including Brazil. Here, two L. major-like strains, one virulent (BH49) and one non-virulent (BH121), were subjected to suppression subtractive hybridization (SSH) technique in order to identify differentially expressed genes. SSH technique identified nine cDNA fragments exhibiting high homology to previously sequenced L. major genes. Five cDNAs (four specific for BH49 and one for BH121) were confirmed by RT-PCR. Among those differentially expressed subtracted genes, some were involved in physiological processes including metabolism, translation and destination of proteins, production of energy, virulence factors and unknown functions. Western-blot analysis confirmed a higher expression level of ß-1,3-galactosyl residues in L. major-like lipophosphoglycan (LPG). This molecular analysis opens the possibility for identification of potential virulence factors not only in different strains, but also in others species of Leishmania.


Assuntos
DNA de Protozoário/genética , Leishmania major/genética , Leishmaniose Cutânea/parasitologia , Animais , Cricetinae , DNA Complementar/genética , Galactosiltransferases/metabolismo , Glicoesfingolipídeos/metabolismo , Humanos , Leishmania major/patogenicidade , Macrófagos/parasitologia , Mesocricetus , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Doenças Negligenciadas/parasitologia , Proteínas de Protozoários/metabolismo , Homologia de Sequência do Ácido Nucleico , Técnicas de Hibridização Subtrativa , Virulência/genética
15.
Diagn Microbiol Infect Dis ; 46(4): 273-8, 2003 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-12944019

RESUMO

Isoenzymes and RAPD (random amplified polymorphic DNA) analysis were used to characterize three Brazilian human isolates of Giardia duodenalis and its clones. The Portland-1 strain (ATCC 30888) was included in the study as a reference pattern. Both methods divided the isolates into two main groups, one represented by the Portland-1 strain, the other constituted by the Brazilian isolates, which, in turn, were divided into 2 subgroups. The dendogram constructed with the RAPD data, using seven primers, revealed a great heterogeneity between Brazilian isolates and the Portland-1 strain. There was no relationship to the clinical characteristics of the isolates. Although a lot of similarity has been observed among Brazilian isolates and its clones, individual polymorphism was detected, which could be related to the clonal reproduction of this protozoan.


Assuntos
DNA de Protozoário/análise , Giardia lamblia/genética , Giardíase/diagnóstico , Isoenzimas/análise , Técnica de Amplificação ao Acaso de DNA Polimórfico/métodos , Animais , Sequência de Bases , Brasil/epidemiologia , Eletroforese em Gel de Amido , Doenças Endêmicas , Feminino , Giardia lamblia/classificação , Giardia lamblia/enzimologia , Giardíase/epidemiologia , Humanos , Masculino , Dados de Sequência Molecular , Estudos de Amostragem , Sensibilidade e Especificidade
16.
Toxicol Lett ; 143(1): 83-92, 2003 Jun 05.
Artigo em Inglês | MEDLINE | ID: mdl-12697384

RESUMO

Isosteviol lactone (LAC), a lactone derivative of the diterpenic acid isosteviol (ISO) was evaluated for its effect on the oxidative metabolism of mitochondria isolated from rat liver. In this model, LAC (1 mM) depressed the phosphorylation efficiency, as shown by the decreased respiratory control coefficient (RCC) and ADP/O ratio. LAC (1 mM) inhibited NADH oxidase (45%), succinate oxidase (34%) and promoted low-level inhibitions on succinate dehydrogenase (13%), succinate-cytochrome c oxide-reductase (23%), cytochrome c oxidase (10%), and NADH dehydrogenase (13%). Glutamate dehydrogenase was also a target for LAC, as it was 85% inhibited by 1 mM LAC. Cyclic voltammetry data showed that LAC, as well as ISO, does not undergo redox reactions under current experimental conditions. LAC (0.05-0.75 mM) inhibited the swelling dependent on the glutamate oxidation, 50% of the effect occurring at 0.5 mM LAC. Swelling supported by KNO(3) and valinomycin was also inhibited over all concentrations used of LAC and ISO, the effect being of a lower intensity for LAC, suggesting that the modification of the structure of ISO by lactonization diminished its interaction with the membrane. This could contribute to attenuation of the toxic effects described for ISO on mitochondrial function, such as those on respiratory chain enzymatic complexes and phosphorylating activity.


Assuntos
Diterpenos do Tipo Caurano , Diterpenos/farmacologia , Lactonas/farmacologia , Mitocôndrias Hepáticas/efeitos dos fármacos , Mitocôndrias Hepáticas/metabolismo , Animais , Diterpenos/síntese química , Diterpenos/química , Eletroquímica , Transporte de Elétrons/efeitos dos fármacos , Eletrofisiologia , Hidrólise , Técnicas In Vitro , Cetonas/química , Lactonas/química , Masculino , Mitocôndrias Hepáticas/enzimologia , Dilatação Mitocondrial/efeitos dos fármacos , Oxirredução , Fosforilação Oxidativa/efeitos dos fármacos , Consumo de Oxigênio/efeitos dos fármacos , Proteínas/metabolismo , Ratos , Ratos Wistar , Stevia/química , Relação Estrutura-Atividade
17.
Biomed Res Int ; 2014: 285607, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-25313356

RESUMO

Entamoeba histolytica is a parasite which presents capacity to degrade tissues and therefore has a pathogenic behavior. As this behavior is not shown by all strains, there have been several studies investigating molecular basis of the cytotoxicity process. Using the suppression subtractive hybridization (SSH) technique, differential gene expressions of two E. histolytica strains, one virulent (EGG) and one nonvirulent (452), have been analyzed with the purpose of isolating genes which may be involved with amoebic virulence. Nine cDNA fragments presenting high homology with E. histolytica previously sequenced genes were subtracted. Of these, four genes were confirmed by RT-PCR. Two coding for hypothetical proteins, one for a cysteine-rich protein, expressed only in the virulent strain, EGG and another one, coding for grainin 2 protein, exclusive from 452 strain. This study provided new insight into the proteins differences in the virulent and nonvirulent E. histolytica strains. We believe that further studies with these proteins may prove association of them with tissue damage, providing new perceptions to improve treatment or diagnosis of the invasive disease.


Assuntos
Entamoeba histolytica/genética , Entamoeba histolytica/patogenicidade , Perfilação da Expressão Gênica , Regulação da Expressão Gênica , Técnicas de Hibridização Subtrativa/métodos , Animais , Células CHO , Cricetinae , Cricetulus , Efeito Citopatogênico Viral/genética , Eletroforese em Gel de Ágar , Eletroforese em Gel de Poliacrilamida , Humanos , Fígado/parasitologia , Masculino , Trofozoítos/fisiologia , Virulência/genética
18.
Pathog Glob Health ; 107(6): 320-4, 2013 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-24091002

RESUMO

The present study evaluates the prevalence of enteroparasitosis in the urban slums of Belo Horizonte, Brazil and the risk of transmitting enteroparasites to the family members of infected individuals. Stool samples were collected and examined at clinical laboratories near each slum. Individuals were identified and classified as positive for parasitosis (IP(+)), and individuals with negative stool tests were classified as negative for parasitosis (IP(-)) and enrolled as control patients. We collected samples from 594 patients, of which 20·2% and 79·8% were classified as IP(+) and IP(-), respectively. In addition, 744 family members (FIPs) effectively participated in the study by providing fecal samples. In total, 1338 participants were evaluated. Of these, 34·6% were tested positive for parasitosis. Blastocystis was the most prevalent parasite, infecting 22·4% of individuals. Among FIPs, the overall prevalence was 46·1%. Of these, 50·6% and 44·7% were classified as FIPs(+) and FIPs(-), respectively. These results showed that IP(+) did not impact the prevalence of infection within the studied communities, not constituting index cases of specific risk behaviors, suggesting that, in fact, these communities are exposed to similar oral-fecal routes of contamination.


Assuntos
Enteropatias Parasitárias/epidemiologia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Brasil/epidemiologia , Criança , Pré-Escolar , Saúde da Família , Fezes/parasitologia , Feminino , Humanos , Lactente , Masculino , Pessoa de Meia-Idade , Áreas de Pobreza , Prevalência , Fatores de Risco , Adulto Jovem
19.
Diagn Microbiol Infect Dis ; 75(2): 160-4, 2013 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-23331963

RESUMO

The chemotherapeutic agents used for the treatment of giardiasis are often associated with adverse side effects and are refractory cases, due to the development of resistant parasites. Therefore the search for new drugs is required. We have previously reported the giardicidal effects of metronidazole (MTZ) and its analogues (MTZ-Ms, MTZ-Br, MTZ-N(3), and MTZ-I) on the trophozoites of Giardia lamblia. Now we evaluated the activity of some giardicidal MTZ analogues in experimental infections in gerbils and its effects on the morphology and ultrastructural organization of Giardia. The giardicidal activity in experimental infections showed ED(50) values significantly lower for MTZ-I and MTZ-Br when compared to MTZ. Transmission electron microscopy was employed to approach the mechanism(s) of action of MTZ analogues upon the protozoan. MTZ analogues were more active than MTZ in changing significantly the morphology and ultrastructure of the parasite. The analogues affected parasite cell vesicle trafficking, autophagy, and triggered differentiation into cysts. These results coupled with the excellent giardicidal activity and lower toxicity demonstrate that these nitroimidazole derivates may be important therapeutic alternatives for combating giardiasis. In addition, our results suggest a therapeutic advantage in obtaining synthetic metronidazole analogues for screening of activities against other infectious agents.


Assuntos
Antiprotozoários/farmacologia , Giardia lamblia/efeitos dos fármacos , Giardíase/parasitologia , Metronidazol/análogos & derivados , Análise de Variância , Animais , Linhagem Celular , Gerbillinae , Giardia lamblia/citologia , Giardia lamblia/ultraestrutura , Concentração Inibidora 50 , Metronidazol/farmacologia , Microscopia Eletrônica de Transmissão , Carga Parasitária , Trofozoítos/citologia , Trofozoítos/efeitos dos fármacos , Trofozoítos/ultraestrutura
20.
Rev Inst Med Trop Sao Paulo ; 55(2): 69-74, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-23563757

RESUMO

Intestinal parasites are an important cause of morbidity and mortality. Immunocompromised individuals may develop more severe forms of these infections. Taking into account the immunity impairment in patients suffering from chronic renal failure (CRF), we will determine the prevalence and associated symptoms of intestinal parasites in these patients. Controls without CRF were used for comparison. Stool samples were collected and processed for microscopic identification of parasites using the Formalin-ether concentration method. For Cryptosporidium diagnosis, the ELISA technique was used. One hundred and ten fecal samples from hemodialysis patients were analyzed, as well as 86 from a community group used as control group. A result of 51.6% of intestinal parasites was observed in hemodialysis patients and 61.6% in the control group. Cryptosporidium and Blastocystis were the most common infections in patients with CRF (26.4% and 24.5%, respectively). Blastocystis was the most common infection in the control group (41.9%), however no individual was found positive for Cryptosporidium. Among the CRF patients, 73.6% were symptomatic, 54.3% of these tested positive for at least one parasite, in contrast to 44.8% in asymptomatic patients (p = 0.38). The most common symptoms in this group were flatulence (36.4%), asthenia (30.0%) and weight loss (30.0%). In the control group, 91.9% were symptomatic, 60.8% of these tested positive for at least one parasite, in contrast to 71.4% in asymptomatic patients (p = 0.703). A significant difference between the two groups was observed with regard to symptoms, with bloating, postprandial fullness, and abdominal pain being more frequent in the control group than in the hemodialysis group (all p < 0.05). Comparing symptomatic with asymptomatic, there was no association in either group between symptoms or the prevalence of parasitic infection, nor with the type of parasite or with multiple parasitic infections. Patients with chronic renal failure are frequent targets for renal transplantation, which as well as the inherent immunological impairment of the disease itself, results in immunosuppression by medication. For this reason, carriers of intestinal parasites with pathogenic potential can develop serious clinical complications influencing the success of transplantation. This fact, coupled with the high prevalence of intestinal parasites and the dissociation between symptoms and infection in CRF patients, suggests that the stool test should be incorporated in routine propedeutics. Furthermore, preventive measures for the acquisition of parasites through the fecal-oral contamination route should be introduced.


Assuntos
Fezes/parasitologia , Enteropatias Parasitárias/epidemiologia , Diálise Renal/estatística & dados numéricos , Adulto , Idoso , Animais , Brasil/epidemiologia , Estudos de Casos e Controles , Estudos Transversais , Feminino , Humanos , Hospedeiro Imunocomprometido , Enteropatias Parasitárias/diagnóstico , Enteropatias Parasitárias/parasitologia , Falência Renal Crônica/parasitologia , Falência Renal Crônica/terapia , Masculino , Pessoa de Meia-Idade , Prevalência
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