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1.
Appl Microbiol Biotechnol ; 108(1): 179, 2024 Jan 27.
Artigo em Inglês | MEDLINE | ID: mdl-38280035

RESUMO

Several COVID-19 vaccines use adenovirus vectors to deliver the SARS-CoV-2 spike (S) protein. Immunization with these vaccines promotes immunity against the S protein, but against also the adenovirus itself. This could interfere with the entry of the vaccine into the cell, reducing its efficacy. Herein, we evaluate the efficiency of an adenovirus-vectored vaccine (chimpanzee ChAdOx1 adenovirus, AZD1222) in boosting the specific immunity compared to that induced by a recombinant receptor-binding domain (RBD)-based vaccine without viral vector. Mice immunized with the AZD1222 human vaccine were given a booster 6 months later, with either the homologous vaccine or a recombinant vaccine based on RBD of the delta variant, which was prevalent at the start of this study. A significant increase in anti-RBD antibody levels was observed in rRBD-boosted mice (31-61%) compared to those receiving two doses of AZD1222 (0%). Significantly higher rates of PepMix™- or RBD-elicited proliferation were also observed in IFNγ-producing CD4 and CD8 cells from mice boosted with one or two doses of RBD, respectively. The lower efficiency of the ChAdOx1-S vaccine in boosting specific immunity could be the result of a pre-existing anti-vector immunity, induced by increased levels of anti-adenovirus antibodies found both in mice and humans. Taken together, these results point to the importance of avoiding the recurrent use of the same adenovirus vector in individuals with immunity and memory against them. It also illustrates the disadvantages of ChAdOx1 adenovirus-vectored vaccine with respect to recombinant protein vaccines, which can be used without restriction in vaccine-booster programs. KEY POINTS: • ChAdOx1 adenovirus vaccine (AZD1222) may not be effective in boosting anti-SARS-CoV-2 immunity • A recombinant RBD protein vaccine is effective in boosting anti-SARS-CoV-2 immunity in mice • Antibodies elicited by the rRBD-delta vaccine persisted for up to 3 months in mice.


Assuntos
Vacinas contra Adenovirus , COVID-19 , Vacinas , Humanos , Animais , Camundongos , Pan troglodytes , ChAdOx1 nCoV-19 , Vacinas contra COVID-19/genética , SARS-CoV-2 , COVID-19/prevenção & controle , Adenoviridae/genética , Vacinação , Anticorpos Antivirais , Anticorpos Neutralizantes
2.
Arch Med Res ; 55(2): 102960, 2024 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-38290199

RESUMO

BACKGROUND: SARS-CoV2 induces flu-like symptoms that can rapidly progress to severe acute lung injury and even death. The virus also invades the central nervous system (CNS), causing neuroinflammation and death from central failure. Intravenous (IV) or oral dexamethasone (DXM) reduced 28 d mortality in patients who required supplemental oxygen compared to those who received conventional care alone. Through these routes, DMX fails to reach therapeutic levels in the CNS. In contrast, the intranasal (IN) route produces therapeutic levels of DXM in the CNS, even at low doses, with similar systemic bioavailability. AIMS: To compare IN vs. IV DXM treatment in hospitalized patients with COVID-19. METHODS: A controlled, multicenter, open-label trial. Patients with COVID-19 (69) were randomly assigned to receive IN-DXM (0.12 mg/kg for three days, followed by 0.6 mg/kg for up to seven days) or IV-DXM (6 mg/d for 10 d). The primary outcome was clinical improvement, as defined by the National Early Warning Score (NEWS) ordinal scale. The secondary outcome was death at 28 d between IV and IN patients. Effects of both treatments on biochemical and immunoinflammatory profiles were also recorded. RESULTS: Initially, no significant differences in clinical severity, biometrics, and immunoinflammatory parameters were found between both groups. The NEWS-2 score was reduced, in 23 IN-DXM treated patients, with no significant variations in the 46 IV-DXM treated ones. Ten IV-DXM-treated patients and only one IN-DXM patient died. CONCLUSIONS: IN-DMX reduced NEWS-2 and mortality more efficiently than IV-DXM, suggesting that IN is a more efficient route of DXM administration.


Assuntos
COVID-19 , Humanos , SARS-CoV-2 , RNA Viral , Tratamento Farmacológico da COVID-19 , Dexametasona/uso terapêutico
3.
Pathogens ; 12(6)2023 Jun 18.
Artigo em Inglês | MEDLINE | ID: mdl-37375530

RESUMO

Recent advances have increased our understanding of the molecular machinery in the cytoskeleton of mammalian cells, in contrast to the case of tapeworm parasites, where cytoskeleton remains poorly characterized. The pertinence of a better knowledge of the tapeworm cytoskeleton is linked to the medical importance of these parasitic diseases in humans and animal stock. Moreover, its study could offer new possibilities for the development of more effective anti-parasitic drugs, as well as better strategies for their surveillance, prevention, and control. In the present review, we compile the results of recent experiments on the cytoskeleton of these parasites and analyze how these novel findings might trigger the development of new drugs or the redesign of those currently used in addition to supporting their use as biomarkers in cutting-edge diagnostic tests.

4.
Planta Med ; 89(2): 148-157, 2023 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-35338475

RESUMO

Plants of Tabernaemontana species have several pharmacological activities including antimicrobial effects. Amoebiasis continues to be a public health problem, with increasing evidence of resistance to metronidazole. In this study, we assessed the effect of the alkaloid fraction of T. arborea root bark and the alkaloids ibogaine and voacangine on the viability and infectivity of Entamoeba histolytica trophozoites. Cultures were exposed to 0.1 - 10 µg/mL for 24, 48 and 72 h, and viability was then determined using a tetrazolium dye reduction assay and type of cellular death analyzed by flow cytometry. Results showed that the alkaloid fraction, but mainly ibogaine and voacangine alkaloids, exhibited potent dose-dependent anti-amoebic activity at 24 h post-exposure (IC50 4.5 and 8.1 µM, respectively), comparable to metronidazole (IC50 6.8 µM). However, the effect decreased after 48 and 72 h of exposure to concentrations below 10 µg/mL, suggesting that the alkaloids probably were catabolized to less active derivatives by the trophozoites. The treatment of trophozoites with the IC50 s for 24 h induced significant morphological changes in the trophozoites, slight increase in granularity, and death by apoptonecrosis. The capacity of T. arborea alkaloids to inhibit the development of amoebic liver abscesses in hamsters was evaluated. Results showed that even when the treatments reduced the number of amoebic trophozoites in tissue sections of livers, they were unable to limit the formation of abscesses, suggesting their rapid processing to inactive metabolites. This work leaves open the possibility of using Tabernaemontana alkaloids as a new alternative for amoebiasis control.


Assuntos
Alcaloides , Amebíase , Ibogaína , Tabernaemontana , Ibogaína/metabolismo , Ibogaína/farmacologia , Metronidazol/farmacologia , Metronidazol/metabolismo , Casca de Planta , Alcaloides/farmacologia , Alcaloides/metabolismo
5.
Diagnostics (Basel) ; 12(7)2022 Jul 05.
Artigo em Inglês | MEDLINE | ID: mdl-35885534

RESUMO

After more than two years, the COVID-19 pandemic is still ongoing and evolving all over the world; human herd immunity against SARS-CoV-2 increases either by infection or by unprecedented mass vaccination. A substantial change in population immunity is expected to contribute to the control of transmission. It is essential to monitor the extension and duration of the population's immunity to support the decisions of health authorities in each region and country, directed to chart the progressive return to normality. For this purpose, the availability of simple and cheap methods to monitor the levels of relevant antibodies in the population is a widespread necessity. Here, we describe the development of an RBD-based ELISA for the detection of specific antibodies in large numbers of samples. The recombinant expression of an RBD-poly-His fragment was carried out using either bacterial or eukaryotic cells in in vitro culture. After affinity chromatography purification, the performance of both recombinant products was compared by ELISA in similar trials. Our results showed that eukaryotic RBD increased the sensitivity of the assay. Interestingly, our results also support a correlation of the eukaryotic RBD-based ELISA with other assays aimed to test for neutralizing antibodies, which suggests that it provides an indication of protective immunity against SARS-CoV-2.

6.
Pharmaceutics ; 15(1)2022 Dec 28.
Artigo em Inglês | MEDLINE | ID: mdl-36678735

RESUMO

Dexamethasone (DXM) and methylprednisolone (MEP) are potent glucocorticoids used to control several inflammatory conditions. Evidence of delayed DXM reaching the central nervous system (CNS) as well as tachyphylaxis and systemic, undesirable side effects are the main limitations of peripheral delivery. Intranasal administration offers direct access to the brain as it bypasses the blood-brain barrier. The Mucosal Atomization Device is an optimal tool that can achieve rapid absorption into the CNS and the bloodstream across mucosal membranes. This study was designed to evaluate and compare the bioavailability of DXM and MEP after intranasal versus intravenous administration. Two open-label, balanced, randomized, two-treatment, two-period, two-sequence, single-dose, crossover studies were conducted, which involved healthy male and female adult volunteers. After intranasal administration, DXM and MEP were detected in plasma after the first sampling time. Mean peak concentrations of DXM and MEP were 86.61 ng/mL at 60 min and 843.2 ng/mL at 1.5 h post-administration, respectively. DXM and MEP showed high absolute bioavailability, with values of 80% and 95%, respectively. No adverse effects were observed. DXM and MEP systemic bioavailability by intranasal administration was comparable with the intravenous one, suggesting that the intranasal route can be used as a non-invasive and appropriate alternative for systemic drug delivery.

7.
Arch Med Res ; 52(2): 143-150, 2021 02.
Artigo em Inglês | MEDLINE | ID: mdl-33160751

RESUMO

The Chinese outbreak of SARS-CoV-2 during 2019 has become pandemic and the most important concerns are the acute respiratory distress syndrome (ARDS) and hyperinflammation developed by the population at risk (elderly and/or having obesity, diabetes, and hypertension) in whom clinical evolution quickly progresses to multi-organ dysfunction and fatal outcome. Immune dysregulation is linked to uncontrolled proinflammatory response characterized by the release of cytokines (cytokines storm). A proper control of this response is mandatory to improve clinical prognosis. In this context, glucocorticoids are able to change the expression of several genes involved in the inflammatory response leading to an improvement in acute respiratory distress. Although there are contradictory data in the literature, in this report we highlight the potential benefits of glucocorticoids as adjuvant therapy for hyperinflammation control; emphasizing that adequate dosage, timing, and delivery are crucial to reduce the dysregulated peripheral-and neuro-inflammatory response with minimal adverse effects. We propose the use of the intranasal route for glucocorticoid administration, which has been shown to effectively control the neuro-and peripheral-inflammatory response using low doses without generating unwanted side effects.


Assuntos
Tratamento Farmacológico da COVID-19 , Glucocorticoides/uso terapêutico , Animais , Citocinas/imunologia , Humanos , Síndrome do Desconforto Respiratório , SARS-CoV-2
8.
Theor Biol Med Model ; 15(1): 18, 2018 11 19.
Artigo em Inglês | MEDLINE | ID: mdl-30449280

RESUMO

BACKGROUND: Taenia solium is the aetiological agent of human taeniasis, pig cysticercosis and human neurocysticercosis, which are serious public health problems, especially in developing countries. METHODS: A mathematical model of the transmission dynamics of taeniasis-cysticercosis is formulated. The model consists of a coupled system of differential equations, which are density-dependent equations for describing the flow of the parasite through the life cycle. The model is hybrid since it comprises deterministic equations with stochastic elements which describe changes in the mean parasite burden and incorporates the overall pattern of the parasites' distribution. RESULTS: Sensitivity and bifurcation analyses were carried out to determine the range of values of the model. The model can reproduce the observed epidemiological patterns of human taeniasis, pig and human cysticercosis. For example, for a wide range of parameter values, the mean intensity of adult worms tends to rapidly stabilize in one parasite per individual host. From this model, we also derived a Susceptible-Infected model to describe the prevalence of infection in humans and pigs. Chemotherapeutic interventions against pig cysticercosis or human taeniasis may reduce rapidly and effectively the mean intensity of human taeniasis, pig cysticercosis and human cysticercosis. This effect can be achieved even if the protective efficacy of the drug is of the order of 90% and the coverage rate is 90%. This means that health in humans infected either with adult worms or cysticerci may be achieved by the application of anthelmintic drugs against pig cysticercosis. However, treatment against human cysticercosis alone, does not influence neither human teniasis nor pig cysticercosis. This is because human cysticercosis infection does not influence the value of the basic reproductive number (Ro). CONCLUSIONS: Even coverage of 100% in the administration of anthelmintics did not eliminate the infection. Then elimination of the infection in all hosts does not seem a feasible goal to achieve by administering only chemotherapeutic interventions. Throughout the manuscript a discussion of our model in the context of other models of taeniasis-cysticercosis is presented.


Assuntos
Anti-Helmínticos/uso terapêutico , Cisticercose/tratamento farmacológico , Cisticercose/transmissão , Modelos Teóricos , Taenia solium/efeitos dos fármacos , Animais , Anti-Helmínticos/farmacologia , Cisticercose/fisiopatologia , Humanos , Estágios do Ciclo de Vida/efeitos dos fármacos , Estágios do Ciclo de Vida/fisiologia , Suínos , Taenia solium/isolamento & purificação , Taenia solium/fisiologia , Teníase/tratamento farmacológico , Teníase/fisiopatologia , Teníase/transmissão
9.
Exp Parasitol ; 172: 23-29, 2017 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-27913109

RESUMO

Taeniasis/cysticercosis caused by the tapeworm Taenia solium is a parasite disease transmitted among humans and pigs, the main intermediate host. The larvae/cysts can lodge in several tissues of the pig, i.e. skeletal muscles and different locations of the central nervous system. The molecular mechanisms associated to tissue preferences of the cysts remain poorly understood. The major public health concern about this zoonosis is due to the human infections by the larval form in the central nervous system, causing a highly pleomorphic and debilitating disease known as neurocysticercosis. This study was aimed to explore the 2DE protein maps of T. solium cysts obtained from skeletal muscles and central nervous system of naturally infected pigs. The gel images were analyzed through a combination of PDQuest™ and multivariate analysis. Results showed that differences in the protein patterns of cysts obtained from both tissues were remarkably discrete. Only 7 protein spots were found specifically associated to the skeletal muscle localization of the cysts; none was found significantly associated to the central nervous system. The use of distinct protein fractions of cysts allowed preliminary identification of several tissue-specific antigenic bands. The implications of these findings are discussed, as well as several strategies directed to achieve the complete characterization of this parasite's proteome, in order to extend our understanding of the molecular mechanisms underlying tissue localization of the cysts and to open avenues for the development of immunological tissue-specific diagnosis of the disease.


Assuntos
Encéfalo/parasitologia , Cisticercose/veterinária , Cysticercus/química , Proteínas de Helminto/análise , Músculo Esquelético/parasitologia , Doenças dos Suínos/parasitologia , Taenia solium/química , Animais , Cisticercose/parasitologia , Cysticercus/isolamento & purificação , Eletroforese em Gel Bidimensional , Sus scrofa , Suínos , Taenia solium/isolamento & purificação
10.
Mol Biochem Parasitol ; 207(2): 61-7, 2016 06.
Artigo em Inglês | MEDLINE | ID: mdl-27234210

RESUMO

Human and porcine cysticercosis is a parasitic disease caused by the larval stage (cysts) of the tapeworm Taenia solium. Cysts may live in several host tissues such as skeletal muscle or brain. We have previously described the presence of host haptoglobin (Hp) and hemoglobin (Hb) in different protein extracts of the T. solium cysts. Here, we report the binding of host Hp and Hb to a number of cyst proteins, evaluated through measuring electrophoretic and light absorbance changes. In the sera obtained from 18 cysticercotic pigs, Hp-Hb complexes were abundant, whereas free Hp was undetectable. In contrast, in the sera from non 18 cysticercotic pigs, Hp-Hb and free Hp were found. In the soluble protein fraction of cysts tissue, free Hp was detected showing a considerable Hb-binding ability, whereas in the vesicular fluid, Hp is mainly bound to Hb. Interestingly, assays carried out with the insoluble fraction of T. solium cysts tissue, showed binding of Hp and Hp-Hb in a saturable way, suggesting the existence of specific interactions. Our results suggested that the parasite can take advantage of the uptaken host Hp and Hb, either free or in complexes, as a source of iron or as a way to modulate the inflammatory response surrounding the T. solium cysts.


Assuntos
Cisticercose/veterinária , Haptoglobinas/análise , Interações Hospedeiro-Parasita , Doenças dos Suínos/imunologia , Doenças dos Suínos/parasitologia , Taenia solium/fisiologia , Animais , Cisticercose/sangue , Cisticercose/parasitologia , Haptoglobinas/metabolismo , Hemoglobinas/análise , Hemoglobinas/metabolismo , Humanos , Músculo Esquelético/parasitologia , Ligação Proteica , Suínos , Doenças dos Suínos/sangue
11.
Biomed Res Int ; 2015: 164980, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26090385

RESUMO

Nanoclays have potential applications in biomedicine raising the need to evaluate their toxicity in in vitro models as a first approach to its biocompatibility. In this study, in vitro toxicity of clinoptilolite and sepiolite nanoclays (NC) was analyzed in highly phagocytic cultures of amoebas and human and mice macrophages. While amebic viability was significantly affected only by sepiolite NC at concentrations higher than 0.1 mg/mL, the effect on macrophage cultures was dependent on the origin of the cells. Macrophages derived from human peripheral blood monocytes were less affected in viability (25% decrease at 48 h), followed by the RAW 264.7 cell line (40%), and finally, macrophages derived from mice bone marrow monocytes (98%). Moreover, the cell line and mice macrophages die mainly by necrosis, whereas human macrophages exhibit increased apoptosis. Cytokine expression analysis in media of sepiolite NC treated cultures showed a proinflammatory profile (INFγ, IL-1α, IL-8, and IL-6), in contrast with clinoptilolite NC that induced lees cytokines with concomitant production of IL-10. The results show that sepiolite NC is more toxic to amoebas and macrophages than clinoptilolite NC, mostly in a time and dose-dependent manner. However, the effect of sepiolite NC was comparable with talc powder suggesting that both NC have low cytotoxicity in vitro.


Assuntos
Silicatos de Alumínio/efeitos adversos , Silicatos de Magnésio/efeitos adversos , Zeolitas/efeitos adversos , Silicatos de Alumínio/uso terapêutico , Animais , Diferenciação Celular/efeitos dos fármacos , Argila , Expressão Gênica/efeitos dos fármacos , Humanos , Interleucina-10/biossíntese , Interleucina-6/biossíntese , Interleucina-8/biossíntese , Macrófagos/efeitos dos fármacos , Silicatos de Magnésio/uso terapêutico , Camundongos , Fator de Necrose Tumoral alfa/biossíntese , Zeolitas/uso terapêutico
12.
Biomed Res Int ; 2015: 760598, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26090442

RESUMO

In a previous study, we demonstrated that oral immunization using Autographa californica baculovirus driving the expression of the Gal-lectin LC3 fragment (AcNPV-LC3) of Entamoeba histolytica conferred protection against ALA development in hamsters. In this study, we determined the ability of AcNPV-LC3 to protect against ALA by the intramuscular route as well as the liver immune response associated with protection. Results showed that 55% of hamsters IM immunized with AcNPV-LC3 showed sterile protection against ALA, whereas other 20% showed reduction in the size and extent of abscesses, resulting in some protection in 75% of animals compared to the sham control group. Levels of protection showed a linear correlation with the development and intensity of specific antiamoeba cellular and humoral responses, evaluated in serum and spleen of hamsters, respectively. Evaluation of the Th1/Th2 cytokine patterns expressed in the liver of hamsters showed that sterile protection was associated with the production of high levels of IFNγ and IL-4. These results suggest that the baculovirus system is equally efficient by the intramuscular as well as the oral routes for ALA protection and that the Gal-lectin LC3 fragment is a highly protective antigen against hepatic amoebiasis through the local induction of IFNγ and IL-4.


Assuntos
Baculoviridae/imunologia , Imunização , Abscesso Hepático Amebiano/imunologia , Vacinas Protozoárias/administração & dosagem , Animais , Antígenos de Protozoários/imunologia , Cricetinae , Entamoeba histolytica/efeitos dos fármacos , Entamoeba histolytica/imunologia , Imunoglobulina G , Abscesso Hepático Amebiano/patologia , Abscesso Hepático Amebiano/prevenção & controle , Vacinas Protozoárias/imunologia
13.
Biomed Res Int ; 2013: 758341, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-23484154

RESUMO

Encystment is an essential process in the biological cycle of the human parasite Entamoeba histolytica. In the present study, we evaluated the participation of E. histolytica Gln6Pi in the formation of amoeba cyst-like structures by RNA interference assay. Amoeba trophozoites transfected with two Gln6Pi siRNAs reduced the expression of the enzyme in 85%, which was confirmed by western blot using an anti-Gln6Pi antibody. The E. histolytica Gln6Pi knockdown with the mix of both siRNAs resulted in the loss of its capacity to form cyst-like structures (CLSs) and develop a chitin wall under hydrogen peroxide treatment, as evidenced by absence of both resistance to detergent treatment and calcofluor staining. Thus, only 5% of treated trophozoites were converted to CLS, from which only 15% were calcofluor stained. These results represent an advance in the understanding of chitin biosynthesis in E. histolytica and provide insight into the encystment process in this parasite, which could allow for the developing of new control strategies for this parasite.


Assuntos
Aldose-Cetose Isomerases/biossíntese , Entamoeba histolytica/enzimologia , Regulação Enzimológica da Expressão Gênica , Proteínas de Protozoários/biossíntese , Interferência de RNA , Trofozoítos/enzimologia , Aldose-Cetose Isomerases/genética , Quitina/biossíntese , Entamoeba histolytica/genética , Humanos , Peróxido de Hidrogênio/farmacologia , Oxidantes/farmacologia , Proteínas de Protozoários/genética , RNA Interferente Pequeno/genética , RNA Interferente Pequeno/metabolismo
14.
Am J Trop Med Hyg ; 88(2): 275-84, 2013 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-23249692

RESUMO

The triatomines vectors of Trypanosoma cruzi are principal factors in acquiring Chagas disease. For this reason, increased knowledge of domestic transmission of T. cruzi and control of its insect vectors is necessary. To contribute to genetic knowledge of North America Triatominae species, we studied genetic variations and conducted phylogenetic analysis of different triatomines species of epidemiologic importance. Our analysis showed high genetic variations between different geographic populations of Triatoma mexicana, Meccus longipennis, M. mazzottii, M. picturatus, and T. dimidiata species, suggested initial divergence, hybridation, or classifications problems. In contrast, T. gerstaeckeri, T. bolivari, and M. pallidipennis populations showed few genetics variations. Analysis using cytochrome B and internal transcribed spacer 2 gene sequences indicated that T. bolivari is closely related to the Rubrofasciata complex and not to T. dimidiata. Triatoma brailovskyi and T. gerstaeckeri showed a close relationship with Dimidiata and Phyllosoma complexes.


Assuntos
Variação Genética , Insetos Vetores/classificação , Triatoma/classificação , Triatoma/genética , Animais , Doença de Chagas/transmissão , Clonagem Molecular , Citocromos b/genética , Insetos Vetores/genética , Insetos Vetores/parasitologia , Repetições de Microssatélites , América do Norte , Filogenia , Filogeografia , Alinhamento de Sequência , Análise de Sequência de DNA
15.
Dalton Trans ; 41(34): 10164-74, 2012 Sep 14.
Artigo em Inglês | MEDLINE | ID: mdl-22722265

RESUMO

A new synthetic pathway was reported to obtain N6 donor ligand 2,9-bis-(2',5'-diazahexanyl)-1,10-phenanthroline (L1) and its coordination compounds of essential divalent metal ions Mn, Fe, Co, Ni, Cu and Zn. Complete characterization of all compounds was done with the conventional techniques. Crystal structures of [NiL1](PF(6))(2) and [ZnL1](PF(6))(2)·H(2)O were also reported. Electrochemical studies have shown an active participation of the aromatic moiety of the ligand in redox reactions. The in vitro tests of the cytotoxic activity against human tumour cell lines HeLa (cervix) and CHP-212 (neuroblastoma) showed that all coordination compounds that involve redox active metal ions exhibit noteworthy antiproliferative activity, superior in all cases to cisplatin. [CuL1](2+) showed the lower IC(50) value in the HeLa cell line with 1.84 µM, meanwhile, [CoL1](2+) showed the lower value in neuroblastoma CHP-212 with IC(50) = 45.28 µM. None of these compounds were active against the SK-N-SH neuroblastoma cell line. In Entamoeba histolytica cultures, remarkable nanomolar IC(50) values were found for [NiL1](2+) and [MnL1](2+) with 60 nM and 80 nM respectively, improving the antiproliferative activity more than 1000 times compared with the first choice drug for clinical treatments of human amoebiasis, metronidazole. On the other hand, a free ligand does not show antiproliferative activity either on human tumor cell lines or on Entamoeba histolytica trophozoites, highlighting the role played by metal ions to produce cytotoxicity in tumor cells and protozoa systems.


Assuntos
Antiprotozoários/química , Complexos de Coordenação/química , Fenantrolinas/química , Elementos de Transição/química , Antiprotozoários/toxicidade , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Complexos de Coordenação/toxicidade , Cristalografia por Raios X , Entamoeba histolytica/efeitos dos fármacos , Células HeLa , Humanos , Conformação Molecular , Fenantrolinas/síntese química
16.
Exp Parasitol ; 130(4): 423-9, 2012 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-22343043

RESUMO

During some estimations of the nuclear DNA content, based on determinations propidium iodide (PI) binding through fluorocytometry for Taenia crassiceps cysticerci, significant variation in the results were found. This initial observation led to a series of experiments designed to explain the variation. These changes could be induced by the diameter of the needles in the syringes used for the mouse to mouse transfer of the cysts. Nuclei from cysts transferred through 27-gauge needles showed 30% less PI staining than those transferred through 21 gauge needles, after 2 months infections. Reduction in PI capture induced by 27-gauge needle was reversible when the cysts were maintained in their mice hosts during 5 months. Moreover, variation in PI binding to cysticercal DNA was also found when comparing parasites grown in male versus female mice. The use of agents that homogenize the chromatin structure during PI staining, allowed demonstrating that variation were entirely due to differences in the chromatin relaxation/compaction. Additional experiments demonstrated that the higher compaction is accompanied by a reduced ability of cysts to grow in the peritoneal cavity of BALB/cAnN mice. Furthermore, proteomic analysis also showed that these changes in chromatin relaxation/compaction resulted in different levels and patterns of protein expression. Our results strongly suggest that chromatin is involved in several well characterized phenomena of the T. crassiceps murine model, and open new avenues for a detailed approach to understand such a complex host-parasite relationship.


Assuntos
Cromatina/metabolismo , Cisticercose/parasitologia , Cysticercus/genética , Proteínas de Helminto/análise , Animais , Cysticercus/fisiologia , DNA de Helmintos/metabolismo , Eletroforese em Gel Bidimensional , Feminino , Citometria de Fluxo , Proteínas de Helminto/química , Ponto Isoelétrico , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Peso Molecular , Proteoma/análise
17.
Am J Trop Med Hyg ; 85(3): 490-7, 2011 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-21896810

RESUMO

The purpose of this study was to conduct an entomological analysis, determination of feeding sources, and molecular identification of triatomines in five communities of the Istmo de Tehuantepec, Oaxaca. The only found species in two of five searched communities (San Mateo del Mar and Tehuantepec City) was Triatoma phyllosoma. Colonization indices were high in both communities. In San Mateo del Mar, the insects were found indoors and in Tehuantepec City in peridomestic areas. The Trypanosoma cruzi infection indices were 2.1% in San Mateo del Mar and 39.4% in Tehuantepec City. This difference could be related to the high numbers of triatomine feeding on hens in the former community. In contrast, in Tehuantepec, dogs were the principal triatomine feeding sources. All nymphs and adults that were genetically analyzed belonged to the species T. phyllosoma. Low levels of genetic variation were found between vectors from both communities.


Assuntos
Doença de Chagas/epidemiologia , Comportamento Alimentar/fisiologia , Insetos Vetores/fisiologia , Triatoma/classificação , Triatoma/fisiologia , Animais , Doença de Chagas/transmissão , Galinhas/sangue , Cães , Humanos , México/epidemiologia , Filogenia , Triatoma/genética , Trypanosoma cruzi/fisiologia
18.
Trends Parasitol ; 27(10): 450-8, 2011 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-21775209

RESUMO

Certain protozoan parasites use survival strategies to reside outside the host such as the formation of cysts. This dormant and resistant stage results from the complex process of encystment that involves diverse molecular and cellular modifications. The stimuli and changes associated with cyst biogenesis are a matter of ongoing studies in human and animal protozoan parasites such as amoeba and Giardia species because blocking every step in the encystment pathway should, in theory, interrupt their life cycles. The present review thoroughly examines this essential process in those protozoan parasites and discusses the possibility of using that information to develop new kinds of anti-parasite specific and life cycle-interrupting drugs, aimed at holding back the dissemination of these infections.


Assuntos
Parede Celular/metabolismo , Entamoeba/crescimento & desenvolvimento , Giardia/crescimento & desenvolvimento , Estágios do Ciclo de Vida , Transdução de Sinais , Aldose-Cetose Isomerases/metabolismo , Animais , Vias Biossintéticas , Quitina/metabolismo , Quitina Sintase/metabolismo , Retículo Endoplasmático/metabolismo , Entamoeba/genética , Regulação da Expressão Gênica , Giardia/genética , Humanos , Polissacarídeos/metabolismo , Proteínas de Protozoários/genética , Proteínas de Protozoários/metabolismo
19.
J Biomed Biotechnol ; 2010: 910375, 2010.
Artigo em Inglês | MEDLINE | ID: mdl-20130782

RESUMO

In order to explore how molecules are linked to the membrane surface in larval Taenia solium, whole cysticerci were incubated in the presence of phospholipase C from Clostridium perfringens (PLC). Released material was collected and analyzed in polyacrylamide gels with sodium dodecyl sulfate. Two major bands with apparent molecular weights of 180 and 43 kDa were observed. Western blot of released material and localization assays in cysticerci tissue sections using antibodies against five known surface glycoproteins of T. solium cysticerci indicated that only one, previously called GP1, was released. Similar localization studies using the lectins wheat-germ-agglutinin and Concanavalin A showed that N-acetyl-D-glucosamine, N-acetylneuraminic, sialic acid, alphamethyl-D-mannoside, D-manose/glucose, and N-acetyl-D-glucosamine residues are abundantly present on the surface. On the other hand, we find that treatment with PLC releases molecules from the surface; they do not reveal Cross Reacting Determinant (CRD), suggesting a novel anchor to the membrane for the glycoprotein GP1.


Assuntos
Clostridium perfringens/enzimologia , Cysticercus/metabolismo , Glicoproteínas de Membrana/metabolismo , Taenia solium/metabolismo , Fosfolipases Tipo C/metabolismo , Animais , Western Blotting , Cysticercus/citologia , Cysticercus/ultraestrutura , Eletroforese em Gel de Poliacrilamida , Histocitoquímica , Glicoproteínas de Membrana/química , Microscopia Eletrônica , Músculo Esquelético/parasitologia , Suínos
20.
Methods ; 49(4): 346-50, 2009 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-19651215

RESUMO

Neurocysticercosis in humans is caused by the tapeworm Taenia solium and generates substantial morbidity in Latin America, Africa and Asia.The life cycle of T. solium includes pigs as intermediate hosts and human beings as definitive hosts. Tapeworm carriers are the main risk factor for acquiring cysticercosis in the household, thus prevention and control programs are being developed. Infected people have no symptoms, therefore are difficult to identify and treat, thus vaccination against the adult tapeworm is an alternative control measure. Since the infection occurs naturally only in human beings, experimental models have been standardized. Hamsters are believed to be good models to study the infection but they have not been properly evaluated for vaccination. Since taeniosis is gained by ingesting pork meat with cysticerci, oral vaccination was evaluated, and given that intestinal immunity is enhanced with adjuvants, cholera toxin was used, because it is one of the most potent adjuvants, in view of the fact that it increases epithelium permeability enhancing entrance of the co-administered unrelated antigens. Recombinant functional T. solium calreticulin was employed for the standardization of the methodology and the evaluation of oral vaccination. Protection was associated with the type of cysticerci and the age of the hamsters used. When reddish bigger parasites were orally introduced in hamsters as challenge, protection was around 40%, while when yellowish small parasites were used, protection increased to 100%, suggesting that the characteristics of cysticerci are determinant. Protection was gained in 9month old hamsters, but not in 3month old animals.


Assuntos
Modelos Animais de Doenças , Taenia solium , Teníase/prevenção & controle , Vacinação/métodos , Vacinas Sintéticas/administração & dosagem , Vacinas Sintéticas/imunologia , Administração Oral , Animais , Cricetinae , Feminino , Humanos , Masculino , Mesocricetus , Suínos , Taenia solium/efeitos dos fármacos , Taenia solium/imunologia , Teníase/imunologia , Teníase/parasitologia
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